Hormonal changes in puberty III: Correlation of plasma dehydroepiandrosterone, testosterone, FSH, and LH with stages of puberty and bone age in normal boys and girls and in patients with Addison's disease or hypogonadism or with premature or late adrenarche. In 104 normal boys, aged 7 to 14 years (bone ages 5 to 15 years), plasma dehydroepiandrosterone (DHEA) rose from 52.7 at 7 years, to 112.0 ng/100 ml at 10 years. A further rise occurred at 12 years (188 ng/100 ml). In relation to the bone age, DHEA increased from a mean plasma level of 31.1 at a bone age of 5 years to 77.1 ng/100 ml at one of 7 years. Further increases were observed with mean values of 163.2 at a bone age of 11 years, and of 221.2 at a bone age of 12 years, with a maximum of 333.4 ng/100 ml at bone ages of 14-15 years. The first significant increase of plasma testosterone (T) was noted at a bone age of 12 years (54.8 ng/100 ml). The major rise of T was preceded by the rise of plasma LH and was accompanied by the rise of plasma FSH. Plasma DHEA and T were also measured in 123 normal girls, ages 6 to 13 years (bone ages 5 to 15 years). DHEA rose significantly from a mean level of 44.7 at 6 years, to 80.9 ng/100 ml at 8 years, with further increases between 9 and 10 years and between 10 and 11 years. In relation to bone age, DHEA increased significantly from a mean plasma concentration of 30.9 at a bone age of 5 years, to that of 58.6 ng/100 ml at 7 years. Further increases were observed with values of 191.1 at a bone age of 10 years and 485.6 ng/100 ml at a bone age of 13 years. The first significant rise of testosterone (T) occurred at 10 years of both chronological and bone age. DHEA rose before the increase of gonadotropins. The major rise of T at a bone age of 10 years occurred concurrently with increases in plasma FSH and LH. Low levels of DHEA were observed in Addison's disease. In hypogonadotropin hypogonadism and in anorchia, DHEA levels were normal, suggesting that DHEA is produced primarily in the adrenal gland. In seven girls with early adrenarche, plasma concentrations of DHEA were in the upper range of normal values, whereas T levels were within the normal range. Conversely in girls with late adrenarche, plasms DHEA was lower than normal but T was within the normal limits. The elevation of DHEA prior to the first signs of puberty suggests that DHEA may play a role in the maturation of the hypothalamic-hypophysealgonadal axis. However, the mechanism that triggers the secretion of DHEA is not known.


Efficacy of phage typing epidemiologically related Staphylococcus epidermidis strains. A total of 118 epidemiologically related Staphylococcus epidermidis strains from hospital patients, staff, and fomites were examined with a provisional set of 18 typing phages. Seventy (59.3%) of these strains were typed using phage concentrations of 100 times routine test dilution. The remainder were nontypable. Thirty-six (30.5%) of the strains were of related phage types, 71/108/275A/459 and 71/108/275A. These latter strains were associated with clinical S. epidermidis endocarditis in patients with prosthetic valve replacements. Ninety-eight strains were characterized by the Baird-Parket biotyping schema. Eighty-three (84.7%) were biotype 1, and the majority (68.4%) of these were resistant to penicillin, ampicillin, methicillin, cephalothin, erythromycin, and clindamycin. Type 71, 71/108/275A/459, 71/108/275A and 71/108/275/459 strains were generally resistant to penicillin, ampicillin, erythromycin, and methicillin, whereas a less consistent resistance pattern was noted among miscellaneous and nontypable strains.


Content and synthesis of glycosaminoglycans in the developing lung. The function of lung is fundamentally linked to the connective tissue composition of the alveolar interstitium. The composition and synthesis of one class of interstitial connective tissue components, the glycosaminoglycans (GAG), was determined in lung parenchyma of rabbits at different stages of development. Parenchymal GAG content ranged between 0.2 and 0.4% (wt/wt) of dry weight, with highest concentration in adult lung. There were significant changes in types of GAG present at different ages. Fetal lungs contained a relatively high proportion of chondroitin 4-sulfate while the GAG in lung parenchyma of older animals was predominantly dermatan sulfate, heparan sulfate, and heparin. Methods were developed for the study of rates of synthesis of GAG by incorporation of [1-14C]glucosamine into lung explants. The rate of synthesis of total GAG per cell increased with development to a maximum in lung from weanling rabbits and fell to low rates of synthesis in mature rabbits. Fetal rabbit lung parenchyma synthesized mostly hyaluronic acid and heparan sulfate, while in weanling rabbit parenchyma hyaluronic acid and chondroitin 4/6-sulfate synthesis was greatest. In mature animals, the rates of synthesis of all types of GAG were relatively low but there was a relatively greater emphasis on synthesis of dermatan sulfate and heparin. These results may have significance in changes in lung function during development and in effects on other connective tissue components.


Immediate-type hypersensitivity to a bean lectin source with commentary on occupational allergy in allergy-immunology laboratory and research workers. A case of acute, severe allergic conjunctivitis is reported in a biochemist who had been grinding Bandeiraea simplicifolia, the source of an anti-B lectin. A crude extract of these beans produced immediate-type skin test reactivity, and reaginic antibodies were demonstrated in her serum. Several other examples of occupationally induced allergy in laboratory and research workers in the field of allergy-immunology are briefly cited. Exposure to potential allergens in a dry, powdered form seems particularly hazardous.


Effects of age on renal function and enzyme activity in male C57BL/6 mice. The activities of renal lactate and malate dehydrogenases, glutaminase, and Na-K-ATPase were determined in aging male C57BL/6 mice. Urine concentrating ability in these mice and renal response to metabolic acidosis were also studied. Total enzyme activities were measured in vitro in tissue homogenates from mice that were 120, 400, 500, 600, 700, and 800 days old. Urine concentrating ability was determined in these mice prior to sacrifice. Lactate and malate dehydrogenase activities decreased between 120 and 700 days with only male dehydrogenase activity increasing between 700 and 800 days. Age did not affect glutaminase or Na-K-ATPase activities and urine concentrating ability was decreased only at 700 days. Both urine ammonia excretion and renal glutaminase activity increased at 120 and 600 days in response to metabolic acidosis. However, only 5 of 12 animals tested at 600 days survived the acid stress for a full 7 days.


Cell-mediated immune responses in vitro. III. Elimination of specific cytotoxic lymphocyte responses by 3H-thymidine suicide. The role of cellular proliferation in the development of cytotoxic lymphocyte (CL) responses in one-way mixed lymphocyte reactions was investigated by using tritiated thymidine of high specific activity to kill proliferating cells. To develop maximum CL responses, responding lymphoid cells must proliferate for approximately 72 hr; thereafter, precursors of CL appear to differentiate into active CL without further proliferation. Different alloantigen-sensitive precursor cell populations participate in the CL responses to each of two sets of stimulating alloantigens. When cells responding to one set of alloantigens were selectively destroyed after incorporating the hot thymidine, the surviving cells retained the capacity to develop a normal CL response to the second set of alloantigens.


Augmentation of proliferation and in vitro production of cytotoxic cells by 2-ME in the rat. Low concentrations (10(-5) to 10(-8) M) of 2-mercaptoethanol (2-ME) greatly enhance the proliferation of allogeneic cells in the rat mixed lymphocyte culture (MLC). Studies were undertaken to determine the mode of action of 2-ME. MLC proliferation can occur in the absence of serum proteins (fetal calf serum, FCS) only if 2-ME is present; however, a synergistic effect is present with FCS plus 2-ME, with a 3-fold increase in 3HTdR incorporation with FCS concentrations as low as 0.1%. Kinetic studies show no shift in the peak of proliferation (92 hr) when comparing cultures with and without 2-ME; however, 2-ME-supplemented cultures have significant 3HTdR uptake at 24 hr, and the peak amount of uptake at 92 hr is two to four times higher. Delayed addition of 2-ME until 92 and 166 hr produces a further increase in 3HTdR uptake, indicating that the entire effect is not expressed at the time of allogeneic recognition. L-ascorbic acid, another reducing agent which lacks sulfhydryl groups, elicits a much lower effect on DNA synthesis than does 2-ME. The cytotoxicity of cells harvested from MLC supplemented with 2-ME is increased without loss of target specificity, whereas the same concentration of 2-ME has no direct effect upon the cytotoxicity assay except at higher concentrations where 2-ME suppresses cytotoxicity.


Cytotoxic T cell activity is strain-specific in outbred mice infected with lymphocytic choriomeningitis virus. The cytotoxic T cell response in outbred mice infected with lymphocytic choriomeningitis virus (LCMV) is strain specific. The same is true for adoptive transfer of fatal LCM disease. The response of individuals within an outbred strain is completely cross-reactive, as shown by using immune lymphocytes and virus-infected macrophage targets from individual mice. Reciprocal exclusion of cytotoxic T cell activity between inbred and outbred mouse strains is the rule, the exception being one strain (H) known to have some C57BL ancestry. Immune T cells from one of 7 H mice specifically lysed LCMV-infected C57BL macrophages. Experiments with inbred mice have shown that only one allele need be shared at either the H-2K or H-2D locus for cytotoxic T cell activity to be manifest. Adoptive transfer protocols may thus be considered in outbred situations, providing that T cells are effective before allograft rejection occurs. Also, the LCMV cytotoxic T cell assay may be useful for determining the degree of H-2 variability in wild mouse populations, as novel H-2 types can be detected and mice need not be congenic.


Suppression of lymphocyte activation by a factor produced by Mycoplasma arginini. Mycoplasma arginini, when grown in broth or in cultures of human lymphoblast cell lines, inhibits activation of lymphocytes by allogeneic cells of mitogens. This inhibition can also be produced by cell free media obtained after the growth of this organism. Inhibition is not species specific and inhibits both B and T cell activation in the mouse.  The inhibitory factor is required during the first 3 days of the mixed leukocyte culture, but has no effect on the latter phase of this reaction when thymidine uptake is greatest. The factor is stable to treatment at 60 degrees, but not 90 degrees, for 30 min.


Generation of cytotoxic T lymphocytes in vitro. VI. Effect of cell density on response in mixed leukocyte cultures. Reexposure of day 14 murine mixed leukocyte culture (MLC) populations to the original irradiated allogeneic stimulating spleen cells has previously been found to result in the ratpid generation of cytolytic T lymphocytes (CTL) associated with a net increase in cultured cell number. Under the experimental conditions used, day 5 MLC cells appeared unable to respond to the allogeneic stimulus. In order to characterize further the development of the potential for anamnestic reactivity during the course of MLC, C57BL/6 spleen cells were incubated with irradiated (1000 rads) DBA/2 spleen cells (primary MLC) for up to 3 weeks. At various time intervals after the onset of the primary MLC, the surviving cells were collected and reexposed, at varying cell concentrations, to irradiated DBA/2 spleen cells (secondary MLC). At daily intervals thereafter, CTL activity was assessed using a quantitative 51Cr-release assay system. A paradoxic effect of responding cell concentration on generation of CTL activity was observed; relatively greater increase in CTL activity was observed as the concentration of responding cells was decreased over a 100-fold range. This effect was more pronounced with responding cells reexposed to antigen after primary MLC for 20 days, but was observed even with normal cells. The apparent unresponsiveness of day 5 MLC cells to alloantigen restimulation could be overcome by simple dilution of responding cells. Cytotoxic activity at the time of restimulation with antigen seems to be a major factor determining the magnitude of the secondary response. Since intact cells bearing alloantigens are required for the generation of CTL in MLC, residual cytotoxic cells reduce the effective antigenic stimulus by destroying stimulating cells. This effect of concentration of responding cells on generation of CTL in MLC complicates interpretation of experiments investigating the role of "inhibitor" and "helper" cell in cell-mediated immune responses occurring in vitro. Under optimal conditions, the highest CTL activity and the largest increase in total cell number was observed 4 days after restimulation of day 10 MLC cells. On a per cell basis, the lytic activity was up to 4 times greater than that observed at the peak of a primary response, and the number of viable cells recovered was nearly 20 times higher than that at the onset. Such secondary MLC are thus a convenient source of lymphoid cells selected primarily on the basis of proliferation induced by alloantigens.


New bacteriophages of Staphylococcus epidermidis. Fifty-six phages designated the U-series and 34 phages designated the Ph-series were carefully examined. The spectrum of activity was established for 183 strains of Staphylococcus epidermidis from different countries and for 258 strains of Staphylococcus aureus. All 90 phages were active against strains of both species, but some differences in sensitivity were noted. Sixteen of the 90 phages were selected for phage typing of S. epidermidis. With use of a routine test dilution of phage (the highest dilution that yields confluent lysis of the propagation strain), 71.6% of 183 coagulase-negative staphylococci tested were sensitive to the phages; 42 phage patterns were observed. The results indicate that phage typing of S. epidermidis can be recommended for epidemiological and ecological purposes.


Ultrastructural identification of extracellular matrix and cell surface components during limb morphogenesis in man. Development of the human hand plate (stages 16-17) has been analyzed with emphasis on differentiation of elements within the extracellular  matrix and the composition of the mesenchymal cell surface. The epithelial-mesenchymal interface contains a basal lamina and a sublaminar matrix exhibiting: (a) collagen fibrils with characteristic 63-64 nm banding: (b) non-banded filaments, 10-15 nm in diameter; (c) ruthenium red-positive particles, 12-15 nm in diameter; and (d) attenuated threads, 3-5-5-0 nm in diameter which inter-connect particles, fibrils, filaments and the basal lamina. Processes of mesenchymal cells penetrate this matrix network. In addition to staining with ruthenium red, components of basal laminae bind to ferritin-conjugated Concanavalin A, greatest binding being localized on the mesenchymal surface of the lamina. Asymmetry of binding is removed by incubation of exposed laminae with trypsin (5 mug/ml). Regional differences in these staining and binding characteristics within the subepithelial matrix have not been observed in the hand plate. However, precartilaginous extracellular zones deep within the plate are notably unstructured in comparison to the sublaminar region. Ruthenium red-positive materials at mesenchymal cell surfaces display sensitivity to testicular hyaluronidase, Pronase and trypsin but resist removal with neuraminidase and EDTA. These features of the substrate in situ may be important in the regulation of mesenchymal cell behavior during limb morphogenesis in man.


The requirement for DNA synthesis and gene expression in the generation of cytotoxicity in vitro. The requirement for cell division and expression of new genes was examined in the primary and secondary mouse mixed leukocyte culture (MLC). Hydroxyurea (HU) was used to block DNA synthesis and cell division, and 5-bromo-2'-deoxyuridine (BUdR) was used to probe for the expression of new cell-specific genes. In the primary MLC, inhibition of DNA synthesis and cell division by HU almost totally suppressed the generation of initial, target-specific cytotoxicity. When HU was washed out of the cultures, cytotoxicity was generated after a lag time approximately equal to the period of treatment with  HU. The rate of development and maximal value of cytotoxicity in HU-reversed cultures was identical to untreated controls, suggesting that the inhibition was not due to a nonspecific lethal effect of the drug. Development of initial cytotoxicity in primary MLC was similarly suppressed by levels of BUdR 25 to 75-fold below the levels of this drug having nonspecific mutagenic effects in lymphocytes, indicating that development of cytotoxicity was also dependent on the expression of a new genetic program. In the secondary MLC, regeneration of both DNA synthesis and cytotoxicity was apparent 12-15 h after re-exposure to initial stimulating antigen. In this reaction, however, generation of cytotoxicity was insensitive to both HU and BUdR. Thus, the cytotoxic program developed in the primary MLC appears to be genetically stable through the production of effector memory cells, and into regeneration of fully cytotoxic memory cells in secondary MLC.


Intercostal muscle biopsy in human neuromuscular disease. Histochemical and electron microscopic studies. External intercostal muscle biopsies were examined histochemically and by electron microscopy. The use of this muscle allowed correlation with physiological and pharmacological studies on the same specimens. Changes observed in musclar dystrophy and motor neurone disease resembled those previously described in biopsied limb muscle and underline the particular usefulness of this preparation in the study of human neuromuscular disease.


Sensory system involvement in infantile spinal muscular atrophy. Nine cases of infantile spinal muscular atrophy were studied post-mortem. Their ages at death ranged from 5 months to 10 years. In all cases severe loss of anterior horn cells in the spinal cord and neurogenic muscular atrophy were characteristic of this disease. In 6 cases there was also loss of myelin in the posterior columns particularly affecting the lumbar contribution. Sensory ganglia, especially from the lumbar region, contained nodules of Nageotte, indicating sensory neuron degeneration. These sensory abnormalities were more severe in the longer surviving cases. It seems possible that sensory neuron degeneration occurs more commonly in Werdnig-Hoffmann disease than has previously been supposed but that it is less severe and develops more slowly than motor neuron degeneration.


Parabiotic reinnervation in normal and dystrophic mice. Part 2. Morphological studies. The technique of parabiotic reinnervation has been used to test directly the neurogenic theory of the aetiology of muscular dystrophy in mice. Dystrophic muscles contain significantly fewer muscle fibres than their normal controls; they also have a much broader spectrum of fibre size because of a much higher proportion of very small fibres and are poorly differentiated into histochemical fibre types. These criteria were used to assess whether there was any amelioration of the dystrophic process in response to the introduction of a normal nerve supply, or whether dystrophic changes were induced in normal muscle reinnervated with a dystrophic nerve. Self-reinnervated normal and dystrophic TA and EDL muscles contained the same numbers of fibres as unoperated controls. The process of parabiosis alone resulted in no changes in normal or dystrophic muscles. In the process of parabiotic reinnervation, the efficiency of the reinnervation process was not affected by the parabiotic state. The parabiotic reinnervation of dystrophic muscle by normal nerve resulted in no significant increase in fibre numbers and the spectrum of fibre sizes was essentially the same as in unoperated dystrophic muscle. The parabiotic reinnervation of normal muscle by dystrophic nerve resulted in a reduction of fibre numbers in only some of the muscles examined. However, the spectrum of fibre diameters remained essentially normal, and the differentiation of the fibres into histochemical fibre types was characteristic of reinnervated normal muscle. There was a marked absence of necrosis or of other histological signs of dystrophy in these muscles. Since there was no positive evidence to show that conversion of normal to dystrophic, or dystrophic to normal muscle occurred under the influence of parabiotic nerve transposition, two alternative conclusions were admissible. Firstly, the influence of dystrophic nerve upon muscle may be operative in fetal or neonatal life and may be irreversible by means of the subsequent introduction of a normal nerve supply. Secondly, the dystrophic state in muscle may be determined by genetic factors independent of nerve supply.


Intraoperative use of Doppler to detect successful obliteration of carotid-cavernous fistulas. Technical note. The authors describe their use of a Doppler flow detector in the treatment of carotid-cavernous fistulas to monitor the ocular bruit, the clinical sign usually used to detect obliteration of the fistula. In seven procedures the Doppler ultrasonic flow detector has yielded satisfactory proof that the flow sounds were no longer audible, and that the surgery was successful.


The responses of muscle spindles in sheep extraocular muscles. 1. Responses from de-efferented muscle spindles in sheep extraocular muscles have been recorded in the Gasserian ganglion using glass microelectrodes. 2. The afferent fibre conduction velocities ranged from 32 to 106 m/sec and most were intermediate. The distribution was unimodal. 3. The afferent responses to ramp stretches and longitudinal vibration of the muscle could not be divided into two groups, but both the dynamic indices and vibration sensitivities of the afferents increased with their conduction velocities. 4. Injection of suxamethonium did not help to classify afferents with intermediate conduction velocity. 5. It is concluded that the afferents from sheep extraocular muscle spindles form a single population whose properties are correlated with their nerve fibre diameters.


Quantitative aspects of ouabain binding to human erythrocyte and cardiac membranes. 1. [3H]ouabain binding to human erythrocyte membranes is a time- and temperature-dependent process. The association of ouabain to the membrane-bound receptor follows second-order kinetics, while the dissociation is a monomolecular reaction. An association rate constant of 4-6 x 10(4) M-1 sec-1 and a dissociation rate constant of 1-4 x 10(-4) sec-1 were measured at 37 degrees C. The dissociation constant calculated from these data agrees with that determined from equilibrium binding experiments. There is only one type of ouabain binding sites with high affinity for the drug as reflected by the low dissociation constant of 0-28 x 10(-8) M. 2. The dissociation constants of the ouabain-receptor complexes from human erythrocyte and cardiac membranes are identical. 3. The maximal number of membrane-bound ouabain binding sites was measured from equilibrium binding experiments as 288 +/- 28 per single erythrocyte. Thus one receptor site corresponds to less than 1 mum2 of the membrane, provided the receptors are diffusely distributed on the surface of the membrane. 4. Neither the maximal number of ouabain receptors nor the affinity for the drug changes with the age or sex of the blood donor. 5. A maximal transport capacity for sodium of 5-6 m-equiv/hr.1. is calculated from the number of receptor sites per erythrocyte and from the turn-over number of the (Na+ + K+)-ATPase.


Responses of neurones in motor cortex and in area 3A to controlled stretches of forelimb muscles in cebus monkeys. 1. The experiments were designed to investigate the effects of longitudinal muscle displacements on neurones of the motor cortex of anaesthetized Cebus monkeys and thus test the hypothesis that signals from muscle spindles may modify motor cortical output. The effects of sinusoidal stretching of the extensor digitorum communis (EDC) at frequencies varying from 6 to 300 Hz and of step and rhomboidal stretches were studied in neurones of the motor cortex. For comparison, neurones of the primary receiving area for low-threshold muscle afferents, cortical area 3a, were also included in this study. Neurones of the motor cortex were subdivided into corticospinal (PT) neurones and non-corticospinal (non-PT) neurones. 2. Threshold stretch amplitudes were clearly higher for neurones of area 4 (PT and non-PT) than for 3a neurones. However, a conspicuous fall in threshold stretch amplitude was observed for all three neurone populations when the frequency of sinusoidal stretching was increased (highest frequency: 300 Hz). A small number of non-PT and PT neurones responded to vibration amplitudes of less than 100 mum and some of these low-threshold cells of area 4 also responded to rhomboidal stretches of 8 mm/sec ramp velocity and 80 mum plateau amplitude. Increasing the stretch amplitude to twice threshold nearly doubled the output magnitude in all three cell types. Neurones of area 3a and non-PT neurones of area 4 had similar latencies, and these were significantly shorter than the latencies of PT neurones tested with trains of high frequency vibration. Dynamic response patterns were observed in all three cell types, but most frequently in 3a neurones. 3. It is concluded that, in Cebus monkeys, signals from both primary and secondary muscle spindle endings from forelimb muscles reach the motor cortex. Under the present experimental conditions, the input from the primaries to the motor cortex was effective only if these spindle receptors were driven maximally by vibratory stimuli. The particularly low probability of stretch-evoked discharges of cortico-spinal neurones in the anaesthetized preparation may be explained by a low gain in transmission from input to output cells of the motor cortex.


A study of subcutaneous rhizolysis in the treatment of chronic backache. Subcutaneous rhizolysis is defined as cutting the nerves to the posterior intervertebral joints. Although a blind procedure, it has proved safe in several centres and provides relief of pain in about two thirds of those who suffer from intractable, persistent backache. Some conditions such as ankylosing spondylitis and previous spinal fusion are clinically unsuitable, but this technique, which is described, is recommended for consideration before all major surgical procedures for back pain are undertaken.


The morphology of follicular development and ovulation in non-human primates. A colony of Macaca fascicularis have been under continuous observation for the past 4 years to ascertain the follicular morphological changes that occur prior to ovulation and during the development of the corpus luteum. Of 609 experimental cycles, laparoscopy was performed at least once in 44-5% of the cycles. Of the 104 cycles where the ovulatory status was definitely known, 89-4% were deemed ovulatory and 10-6% anovulatory. The presence or absence of ovulation in the previous cycle did not have an effect on the cycle length either for the total cycles or when analysing only cycles over 28 days. Similarly, the occurrence of two consecutive ovulations (in consecutive cycles) on the same vs. opposite ovaries did not have a significant effect on the cycle length. Neither laparoscopic stress or anaesthesia effected the normal cyclicity of the animals. The characteristic changes in follicular morphology are most clearly defined in M. fascicularis. In this species the 24 of 36 hr prior to ovulation are accompanied by discrete changes which occur in a fixed sequence, allowing one to predict the time of ovulation with reasonable accuracy. In S. sciureus ovulation is preceded by extensive bulging at the follicular apex and haemorrhaging at the base of the follicle. Due to this haemorrhaging post-ovulatory follicles in S. sciureus are generally more easily discernible than in fascicularis. The formation of clear areas (stigma) is not as evident in either S. sciureus or G. senegalensis as in the macaque. Actual ovulation has been observed four times in M. fascicularis and twice in S. sciureus.


Reinstitution of gold therapy in rheumatoid arthritis after mucocutaneous reactions. Gold therapy was successfully reinstituted in 28 of 30 patients with rheumatoid arthritis who had previously developed toxic, mucocutaneous reactions. At least six weeks after the reactions had completely cleared, patients were given 1 to 10 mg of gold sodium thiomalate (Myochrysine). This dose was increased every two to four weeks. An initial dose of 1 or 2 mg, with an interval of at least three weeks between injections, increasing not more than5 mg at each injection, is recommended.


Investigation of sacroiliac disease: Comparative evaluation of radiological and radionuclide techniques. An attempt has been made to improve diagnostic precision in a group of diseases associated with inflammation of the sacroiliac joints, by using a 99mTechnetium stannous pyrophosphate bone scan. Inflammation of these joints is associated with osteoblastic activity and is reflected by an increase in the uptake of radionuclide, which can be precisely quantitated. The uptake was markedly above the range of normal in patients with active ankylosing spondylitis (AS), and also in a number of patients with possible AS, psoriasis, and Reiter's syndrome. However, patients with Grade 4 radiological changes of the sacroiliac joints frequently had normal scans. This technique may be useful in the early diagnosis of sacroiliitis, and the nosological implications of the changes in patients with Reiter's syndrome and psoriasis are of interest.


Organ culture of adult human articular cartilage. I. The effect of hyperoxia on synthesis of glycosaminoglycan. Adult human articular cartilage was maintained in organ culture for eight days in 20 per cent and 95 per cent oxygen. The higher oxygen tension proved to be lethal to the cartilage, whereas the mean number of surviving chondrocytes in 20 per cent oxygen was 91 per cent of the total. Specimens were cultured  with radioactive sulfate (Na235SO4) for two days and the uptake of sulfate (DPM) at 95 per cent oxygen was significantly lower than at 20 per cent oxygen. There was a positive correlation between DPM and uronic acid, and between DPM and DNA (p less than 0.001), but not between DPM and dry weight.


Organ culture of adult human articular cartilage. II. The differential rate of glycosaminoglycan synthesis in layers of articular cartilage matrix. Adult human articular cartilage was maintained in culture for three days and labelled with 35SO4 for the first 48 hours. The explants were sectioned horizontally on the cryostat, and the sections from the top, middle, and deep layers analyzed  separately. Autoradiographs were prepared from every twentieth section. The DNA content in each of the three layers was of the same order, but the sulfate uptake and uronic acid content were considerably reduced at the surface. It is suggested that the chondrocytes synthesize matrix appropriate to the layer in which they lie.


Partial purification and properties of a bacteriophage T7 inhibitor of the host exonuclease V activity. Infection of Escherichia coli with bacteriophage T7 results in an inhibition of the host exonuclease V (recB, C DNase) activity. This inhibition is not observed when cells are infected in the presence of chloramphenicol or with a gene 1 mutant. The protein responsible for the inhibition of exonuclease V has been partially purified from T7-infected cells. The protein which does not possess nuclease or ATPase activity can inhibit all nucleolytic activities associated with exonuclease V. The protein does not, however, inhibit the DNA-dependent ATPase activity associated with exonuclease V. The inhibitory protein has a molecular weight of about 12,000, as determined from sedimentation analysis in glycerol gradients.


Turner-mongolism polysyndrome. Review of the first eight known cases. To date, a total of eight cases of the Turner-mongolism polysyndrome have been recognized. The clinical manifestations included retarded growth (resulting in a small and infantile appearance), with shield-like chest, poorly developed breasts, absent body hair, brachycephaly, short neck with foldings and low hairline, oblique eyes with epicanthal folds, squat nose, scrotal or normal tongue, abnormal hard palate (high or cleft), short hands and feet, frequent cubitis valgus, normal clitoris (may be either hypoplastic or peniform), mental retardation, and the XO/G+ karyotype, mosaic for XO in most instances. At this time, a single cause for all cases of the double aneuploidy is not known.


Magnetorheography; calculation of blood flow from surface induced potentials. This research is an attempt to measure blood flow without doing any hurt to the body. When the body is placed in a high magnetic field the blood-flow-induced electromotive force (EMF) can be detected with the surface electrodes using the same principle as an electromagnetic flow-meter. This blood flow measurement is named magnetorheography (its recording: MRG). A theoretical analysis of the potential in the tissue shows that blood flow can be calculated from the surface-induced EMF (MRG) without measuring the radius and the depth of the vessel from the skin surface, when the surface is fairly flat in the vicinity of the vessel. A model experiment verified the theory. In order to apply the theory to the measurement in an in situ situation, a structural coefficient was introduced which is related to the external shape, internal tissue compositions and their impedance of the body. Using such a coefficient the flow calculated from MRG, by an equation including the coefficient, agreed with the flow actually recorded. In experiments on the thighs of 8 dogs weighing 7 to 17 Kg, MRG proved to be proportional to arterial flow and the coefficients were approximately constant. These results indicate that blood flow may possibly be estimated by the calculation from MRG.


Effect of diuretics on ion transport of kidney cortex mitochondria. III. Species difference in calcium accumulation and in ethacrynic acid effect. Effect of inorganic phosphate (4 X 10(-3) M) on Ca++-accumulation was examined in kidney cortex mitochondria. Ca++-accumulation of rat kidney cortex mitochondria was slightly influenced by inorganic phosphate. On the other hand, dog kidney cortex mitochondria did not accumulate calcium from the incubation medium until the inorganic phosphate had been added. Ca++-accumulation of rabbit kidney cortex mitochondria was markedly stimulated by inorganic phosphate. When ethacrynic acid was added to the reaction medium in the absence of inorganic phosphate, Ca++-accumulation of rat kidney cortex mitochondria was depressed and the decrease in calcium content of rabbit and dog kidney cortex mitochondria was enhanced. In the presence of inorganic phosphate, the inhibition of Ca++-accumulation by ethacrynic acid was observed only on dog kidney cortex mitochondria. Subsequently, the effect of inorganic phosphate (4 X 10(-4) M) and ethacrynic acid (1 X 10(-4) M on Ca++-ATPase was examined in kidney cortex mitochondria. The low concentration of inorganic phosphate (4 X 10(-4) M) activated Ca++-ATPase of kidney cortex mitochondria in all animal species. The greatest activation of Ca++-ATPase occurred in rabbits, but the activity of the enzyme was lower than that in rats and dogs. Inhibition of Ca++-ATPase by ethacrynic acid was depressed by the addition of inorganic phosphate in kidney cortex mitochondria of experimental animals. Ca++-accumulation may be regulated through the stimulating effect of inorganic phosphate and the inhibitory effect of ethacrynic acid on Ca++-ATPase in kidney cortex mitochondria. Species difference in ethacrynic acid effect on Ca++-accumulation in kidney cortex mitochondria of rats, rabbits and dogs is discussed.


Effects of various drugs on serum free and total tryptophan levels and brain tryptophan metabolism in rats. Various drugs known to bind to serum albumin were examined to determine whether or not they influenced the level of free tryptophan in serum in vitro and in vivo. Possible relationships between the serum free tryptophan level and serotonin (5-HT) synthesis in the brain and the hypothermic effects of these drugs were investigated. Of the drugs examined, sodium salicylate, sodium benzoate and indomethacin caused a significant increase in the concentration of serum free tryptophan and stimulated the synthesis of 5-HT in the brain. Hypothermia induced by salicylate and indomethacin was potentiated by pretreatment with pargyline, a monoamine oxidase inhibitor. Administration of benzoate did not cause any change in body temperature, but after pargyline a hypothermia did occur. However, pretreatment with parachlorophenylalanine, an inhibitor of 5-HT synthesis, did not influence the hypothermia induced by salicylate and indomethacin. Relationship between the hypothermic effect and the increase of 5-HT synthesis in the brain after a large dose of salicylate and indomethacin is discussed.


[Angiocardiographic evaluation of left atrial volume in patients with mitral valve defect]. The cine- and angiocardiographic method permits a quantitative evaluation of the left atrial volume. The maximum volume of the left atrium was found to increase along with the enhancement of mitral regurgitation. Mitral insufficiency is not always accompanied by a "systolic expansion" of the left atrium. The size of the left atrium chamber is mainly determined by the state of the myocardium, and does not much depend on the elevation of the intra-atrial pressure. In cases of mitral defects the increase of the left atrial volume and the changes in the pulse gradient of the left atrial volumes are a compensatory mechanism in the work of the left atrium.


[Effect of sensitization on the characteristics of development of experimental atherosclerosis]. Tests were set up on 99 rabbits to study the influence of sensitization with the vascular "atherosclerotic" antigen on the features specific for the development of experimental atherosclerosis. The results obtained bear witness to a variable effect of sensitization that depends upon the timing of the latter. Sensitization practised concurrently and right from the beginning of feeding the rabbits on cholesterol introduced some elements of prophylaxis of the experimental atherosclerosis. On the other hand, sensitization instituted 3 months after starting the cholesterol feeding of the rabbits resulted in the aggravation of the disease, manifesting itself not in the extension of the area of the atherosclerotic lesion of the aorta, but in an increased incidence of myocardial infarctions.


[Early electrocardiographic signs of chronic pulmonary heart disease]. Early electrocardiographic diagnosis of chronic cor pulmonale was attempted in the study of 312 patients with chronic pneumonia and in 251 normal individuals. In comparing the mean values of several ECG indices in chronic pneumonia, Stage 1, some signs with significant differences were revealed (amplitude of the PaVF wave, ratio of the hight of the PII and TII Waves, are of the PII wave). These signs appeared to be most informative. The following groups were singled out: 38 patients with chronic pneumonia, Stages 1 and 2 with signs of bronchospasm, and 53 persons with the same stages of the disease, but free of bronchial patency disorders. The ECG of these groups of patients was evaluated by means of the tables of electrocardiographic diagnosis of right atrial hypertrophy. The signs of overloading of the right atrium were revealed with statistical significance more often in patients with bronchospasm. The possibility of using this table for revealing early electrocardiographic signs of chronic cor pulmonale was also established; these signs include the amplitude of the PII wave greater than 2 mm and PaVF 1.5 mm, ratio of the hight of the PII and TII waves less than 0.75, the are of the PII wave greater than 0.09.


[Electro- and phonocardiographic changes in cases of a common arterial trunk]. Analysing the data of the examination of 27 patients with a common arterial duct the authors studied the phono- and electrocardiographic changes observed in this cardiac defect. The probable phono- and electrocardiographic signs of this pathology are described the criteria that permit to suspect the presence of a common arterial duct are presented.


Efficacy of lincomycin and spectinomycin on canine pathogens. Lincomycin (L), spectinomycin (S), and both (L-S) were evaluated in vitro and vivo against canine pathogens. In vitro, a wider spectrum of antimicrobial activity was achieved with L-S than with either L or S. The therapeutic effects of the antibiotics were investigated in 36 dogs inoculated with a mixture of the microbal agents. Tonsillitis, conjunctivitis, laryngitis, abscess formation, and necropsy findings were measured. The L-S combination was effective for the treatment of tonsillitis, conjunctivitis, laryngitis, and pneumonia in dogs.


Selective decontamination of the digestive tract of pregnant rabbits: a method for producing Enterobacteriaceae-free rabbits. Selective elimination of the Enterobacteriaceae species from the microflora of pregnant rabbits was achieved by the use of nalidixic acid and cotrimoxazole. Animals born under Enterobacteriaceae-free conditions remained so as long as adequate isolation conditions were maintained.


Mesangial lesions and focal glomerular sclerosis in the aging rat. The pathogenesis of focal glomerular sclerosis (FGS) and its relation to proteinuria and idiopathic nephrotic syndrome are unknown. Urine protein excretion in Sprague-Dawley rats increased with age. Fifty per cent of 12-month and 90 per cent of 24-month-old animals were proteinuric (greater than 20 mg. per day). Heavily proteinuric old rats manifested biochemical changes characteristic of nephrotic syndrome without significant loss of renal function. Three-month, 6-month, and nonproteinuric 12-month-old animals had mesangial deposits of IgM in occasional lobules of some glomeruli and slight mesangial hyperplasia. Four proteinuric 12-month-old rats had diffuse 4+ deposits of IgM in the mesangium of most glomeruli, basement membrane thickening and epithelial cell foot process fusion without FGS. The mesangial IgM deposits eluted in acid buffer and did not fix complement. Six proteinuric 12-month-old rats had focal and segmental areas of glomerular sclerosis with adhesions to Bowman's capsule, foamy cells, intraluminal eosinophilic deposits and capillary wall wrinkling and collapse. These lesions were more advanced in 24-month-old animals. Nonproteinuric 24-month-old rats did not have detectable FGS. Mesangial uptake of colloidal carbon was normal in proteinuric and nonproteinuric animals without FGS. Mesangial uptake of colloidal carbon was normal in proteinuric and nonproteinuric animals without FGS and reduced in proteinuric animals with FGS. In the aging rat the development of proteinuria and mesangial IgM deposition apparently precede development of a focal sclerotic glomerular lesion with histologic and ultrastructural features similar to FGS in man. The generalized impairment of mesangial phagocytic function in proteinuric rats with FGS suggests that this lesion may result from mesangial overload and dysfunction consequent to the persistent increase in glomerular permeability and proteinuria.


Exploring sites on mitochondrial ATPase for catalysis, regulation, and inhibition. Evidence is presented that mitochondrial ATPase has two types of sites that bind adenine nucleotides. The catalytic site, C, binds the substrates ATP, GTP, or ITP and the inhibitor guanylyl imidodiphosphate (GMP-PNP). A second type of site, R, binds ATP, ADP, adenylyl imidodiphosphate (AMP-PNP), and the chromium complexes of ATP or ADP. All of these substances binding to the R site inhibit the hydrolysis of ATP in a competitive manner; their inhibition of hydrolysis of ITP and GTP is noncompetitive. GMP-PNP inhibits oxidative phosphorylation in submitochondrial particles but AMP-PNP does not. The localization on mitochondrial membranes of sites for the binding of various antibiotics that inhibit oxidative phosphorylation is discussed.


Interaction of homogeneous mitochondrial ATPase from rat liver with adenine nucleotides and inorganic phosphate. Mitochondrial ATPase from rat liver mitochondria contains multiple nucleotide binding sites. At low concentrations ADP binds with high affinity (1 mole/mole ATPase, KD = 1-2 muM). At high concentrations, ADP inhibits ATP hydrolysis presumably by competing with ATP for the active site (KI = 240-300 muM). As isolated, mitochondrial ATPase contains between 0.6 and 2.5 moles ATP/mole ATPase. This "tightly bound" ATP can be removed by repeated precipitations with ammonium sulfate without altering hydrolytic activity of the enzyme. However, the ATP-depleted enzyme must be redissolved in high concentrations of phosphate to retain activity. AMP-PNP (adenylyl imidodiphosphate) replaces tightly bound ATP removed from the enzyme and inhibits ATP hydrolysis. AMP-PNP has little effect on high affinity binding of ADP. Kinetics studies of ATP hydrolysis reveal hyperbolic velocity vs. ATP plots, provided assays are done in bicarbonate buffer or buffers containing high concentrations of phosphate. Taken together, these studies indicate that sites on the enzyme not directly associated with ATP hydrolysis bind ATP or ADP, and that in the absence of bound nucleotide, Pi can maintain the active form of the enzyme.


Purification and properties of membrane-bound coupling factor-latent ATPase from Mycobacterium phlei. The membrane bound coupling factor-latent ATPase was solubilized from the membrane vesicles of Mycobacterium phlei by using 0.25 M sucrose or low ionic strength buffer. Purification of the solubilized enzyme by use of Sepharose-ADP conjugate gel yielded a homogenous preparation of latent ATPase which was purified about 216-fold in a single step with an 84% yield. The enzyme exhibits a specific activity of 39 mumoles of ATP hydrolyzed per min per mg protein. The purified enzyme exhibits coupling factor activity. Electrophoresis in two dissociating solvent systems indicates that the enzyme contains at least three major polypeptides of molecular weights 56,000, 51,000, and 46,000 daltons, and two minor polypeptides of 30,000 and 17,000 daltons. Equilibrium binding studies of ADP with purified coupling factor-latent ATPase reveal the presence of two nucleotide binding sites per molecule with an apparent Ka of 8.1 X 10(-5) M. By use of affinity chromatography, another latent ATPase has been isolated from the solubilized enzyme, which does not exhibit coupling factor activity.


Partial purification of active delta and epsilon subunits of the membrane ATPase from escherichia coli. We have partially purified active delta and epsilon subunits of the E. coli membrane-bound Mg2+-ATPase (ECF1). Treating purified ECF1 with 50% pyridine precipitates the major subunits (alpha, beta, and gamma) of the enzyme, but the two minor subunits (delta and epsilon), which are present in relatively small amounts, remain in solution. The delta and epsilon subunits were then resolved from one another by anion exchange chromatography. The partially purified epsilon strongly inhibits the hydrolytic activity of ECF1. The epsilon fraction inhibits both the highly purified five-subunit ATPase and the enzyme deficient in the delta subunit. The latter result indicates that the delta subunit is not required for inhibition by epsilon. By contrast, two-subunit enzyme, consisting chiefly of the alpha and beta subunits, was insensitive to the ATPase inhibitor, suggesting that the gamma subunit may be required for inhibition by epsilon. The partially purified delta subunit restored the capacity of ATPase deficient in delta to recombine with ATPase-depleted membranes and to reconstitute ATP-dependent transhydrogenase. Previously we reported (Biochem, Biophys. Res. Commun. 62:764 [1975]) that a fraction containing both the delta and epsilon subunits of ECF1 restored the capacity of ATPase missing delta to recombine with depleted membranes and to function as a coupling factor in oxidative phosphorylation and for the energized transhydrogenase. These reconstitution experiments using isolated subunits provide rather substantial evidence that the delta subunit is essential for attaching the ATPase to the membrane and that the epsilon subunit has a regulatory function as an inhibitor of the ATPase activity of ECF1.


Studies of substructure and tightly bound nucleotide in bacterial membrane ATPase. Highly purified preparations of Streptococcus faecalis ATPase contain a similar but inactive protein detected by prolonged polyacrylamide gel electrophoresis. The inactive protein appears to arise by proteolytic cleavage of the major subunits in the enzyme. By use of a new technique, subunit analysis in SDS gels was performed on the enzyme band and the inactive protein band excised from a polyacrylamide gel after electrophoresis. The results indicated that the ATPase has the composition alpha3beta3gamma in which alpha = 60,000, beta = 55,000, and gamma = 37,000 daltons. The inactive protein appears to have the composition (f)6 in which f = 49,000 daltons. There is also evidence that the enzyme band contains some slightly modified forms of the ATPase, such as alpha3beta2 (f)gamma. The inactive protein lacks the capacity for tight nucleotide binding. Our experiments show that the tight ATPase-nucleotide complex formed in S. faecalis cells (the endogenous complex) behaves differently from the tight complex formed in vitro (the exogenous complex). We prepared a doubly labeled complex containing endogenous 32P-labeled ADP and ATP and exogenous 3H-labeled ADP. We observed that the addition of free nucelotide to the doubly labeled ATPase displaced the exogenous bound ligand from the enzyme but not the endogenous bound nucleotide. We suggest that the displaceable and nondisplaceable forms of the tight ATPase-nucleotide complex correspond to two different conformational states of the enzyme.


The possible role of tightly bound adenine nucleotides in oxidative and photosynthetic phosphorylation. The tightly bound nucleotides of the beff-heart mitochondrial ATPase are released during cold inactivation followed by ammonium sulfate precipitation. During incubation at 0 degrees C the sedimentation coefficient (S20W) of the ATPase first declines from 12.1S to 9S. Prolonged incubation or precipitation with ammonium sulfate leads to dissociation of the 9S component into subunits with S20W of 3.5S. The 9S component still bears bound nucleotides which exchange more extensively and rapidly with added nucleotides than those bound to the active 12.1S component. The bound nucleotides are lost when the 9S form dissociates into the smaller subunits. Thus, firm binding of nucleotides is a property of the quarternary structure of the enzyme. The exchangeability of the nucleotides bound to the ATPase of chloroplast membranes is greatly increased in membranes illuminated in the presence of pyocyanine. Pi can exchange into both the beta and gamma positions of the bound nucleotides when the membranes are energized in the presence of Mg2+. The exchange of the nucleotides and the incorporation of Pi are insensitive to the inhibitor Dio-9 but are inhibited by the uncoupler S13. This inhibition by S13 parallels that of the inhibition of photosynthetic phosphorylation. These findings are discussed with regard to our hypothesis that electron transfer causes release of preformed tightly bound ATP from the ATPase by inducing a conformational change.


Crosslinking studies on the Ca2+, Mg2+-activated ATPase of Escherichia coli. Crosslinking of membrane proteins of Escherichia coli with dithiobis (succinimidyl propionate) (DSP) resulted in loss of several enzyme activities including the Ca2+, Mg2+-activated ATPase. This enzyme was crosslinked by DSP to the membrane and was not released by dialysis at low ionic strength in the absence of dithiothreitol which could cleave the crosslinking group. DSP inactivated both phosphohydrolase and coupling activities of the solubilized ATPase. Loss of hydrolytic activity could be correlated with the extent of reaction of the alpha and/or beta subunits of the enzyme. The loss of coupling activity appeared to be associated with modification of the gamma and/or delta subunits.


Impaired cell-mediated immunity in patients with cancer. The ability of peripheral blood lymphocytes to respond in vitro to phytohemagglutinin (PHA) and to allogeneic cells in mixed leukocyte reaction (MLC) was studied in 85 patients with cancer and in 50 healthy controls. The effect produced by sera from cancer patients on in vitro lymphocyte blastogenesis was tested on autologous cells and on homologous cells from a constant panel of 10 normal volunteers. Patients with cancer showed a distinct deficiency of cellular immune responsiveness reflected in a stage-related impairment of PHA and MLC reactivity. This deficiency seems at least partially attributable to the presence of lymphocyte depressive factors in cancer sera, since such sera reduced the reactivity of both autologous and normal homologous lymphocytes to a level that was significantly lower than that found in the presence of pooled normal serum. The inhibitory activity of cancer sera was directly related to the extent of the neoplasia.


The significance of coronary arterial stenosis during cardiopulmonary bypass. Myocardial infarction may develop during an uneventful open-heart operation. In order to better understand this complication, we undertook an experimental study. The left circumflex coronary artery of 20 dogs was narrowed to 50 per cent of its area by a metal screw clamp to produce a localized coronary stenosis. Regional myocardial perfusion in the distribution of both the stenotic circumflex and normal left anterior descending (LAD) coronary arteries was measured by injection of a radioactive-labeled microsphere (15 +/- 5 mu). Circumflex coronary artery flow was measured with an electromagnetic flow probe. An epicardial electrogram was recorded in the distribution of the left circumflex. Measurements of regional myocardial perfusion, circumflex flow, and the epicardial electrogram were performed in each animal during the control (prebypass) state and during cardiopulmonary bypass with a beating and fibrillating ventricle. Half the animals had cardiopulmonary bypass performed at 50 mm. Hg perfusion pressure and half at 100 mm. Hg. The animals were put to death at the end of the study, and the hearts were sectioned, weighed, and counted. A cast was made of the stenotic circumflex coronary artery, the degree of stenosis is measured, and the per cent area stenosis calculated. The study showed that the effect of a 50 per cent coronary stenosis in reducing distal flow is apparent only during cardiopulmonary bypass at reduced pressure. The mechanism whereby a myocardial infarction develops during cardiopulmonary bypass could evolve from the development of a "critical" stenosis out of a mild-moderate one at a reduced perfusion pressure during cardiopulmonary bypass.


A new method for temporary left ventricular bypass. Preclinical appraisal. Surgical patients who cannot be weaned from cardiopulmonary bypass during operation or who develop balloon-dependent left ventricular failure postoperatively are now considered unsavable. However, in those with potentially reversible ventricular dysfunction, recovery might be possible if an improved means of temporary circulatory support were available. Towards this end, a pneumatically actuated, left ventricular assist pump was developed and evaluated in 20 consecutive calf experiments. The device, containing a flexible polyurethane pumping chamber, was positioned on the chest wall and connected to the left ventricular apex and descending thoracic aorta by two Dacron valved conduits (xenograft valves). All animals survived a 14 to 30 day pumping interval, and 7 underwent successful removal of the device by division of the Dacron conduits below skin level. As a prelude to human investigation, pumps were implanted during a series of routine autopsies through midline sternotomy incison. The device was positioned on the right anterolateral chest wall, with two valved conduits traversing the mediastinum to connect the pump to the left ventricular apex and ascending aorta.


A new method for prosthetic valve replacement in congenital aortic stenosis associated with hypoplasia of the aortic valve ring. The smallest commercially available prosthetic valve has an external diameter of 17 mm. Therefore, prosthetic valve replacement has been unfeasible in cases in which the diameter of the aortic valve ring is smaller than this size. A new operative method was used on 2 patients with congenital aortic valve lesions associated with hypoplasia of the valve ring. The procedure consists of a longitudinal incision in the aortic septum placed in the midportion of the two coronary ostia, a vertical incision in the outflow tract of the right ventricle to join the septal incision, prosthetic aortic valve replacement, and patch reconstruction of the outflow tracts of both ventricles by means of two layers of a fusiform Dacron patch. A 23-year-old woman with congenital aortic stenosis associated with hypoplattic aortic valve ring underwent prosthetic placement of the aortic valve and is well 1 year after the operation. Our other patient, a 3-month-old infant with congenital aortic regurgitation, died on the first postoperative day because of postperfusion lung. However, her hemodynamic status had been good in the immediate postoperative period.


[SISI-Test and functional hearing loss (author's transl)]. The original SISI test (Jerger, Shedd and Harford) is described in German language. Modifications of various authors are cited. The results of SISI test with 16 persons demanding monetary compensation for hearing loss are tabulated. 18 ears seemed free from non-organic overlay. 12 of those ears gave SISI scores more than 70%. Functional discrepancies wer observed in 13 ears. Only in one of them a maximum SISI score of 25% was presented. All other non-organic ears gave lower ratings, 0% in most cases. Increments of 5 dB were not responded with 5 ears. In one case of unilateral non-organic loss SISI scores were high in the normal and low in the aggravating ear. It is suggested therefore, that low SISI ratings should arise suspicion of non-organic problems, besides of retrocochlear site.


[The diagnosis of conductive hearing loss by tympanometry (author's transl)]. The use of electronic impedance measuring instruments to determine the acoustic impedance of the ear drum is very helpful in the diagnosis of middle ear diseases. Fluid filled middle ear spaces, otosclerosis and tympanosclerosis can be differentiated easily. In addition tube and facial nerve testing is possible. All tests can be done in less than 10 minutes for both ears.


[Unilateral reversible high-tone loss after rubella in adults (author's transl)]. Report on a patient age 18 in whom on the eighth day after a new, seriologically proved rubella infection a unilateral transitory, mediobasal laesio auris internae occurred without any vestibular irritation. Reference is made to the importance of carrying out an examination for a possibly viral genesis as soon as possible after the occurence, by which any type of sudden deafness is understood. Discussion on the development of the hearing defect on the basis of the available literature, whereby an endolymphatic labyrinthitis may be assumed.


[Different forms of deafness in Treacher - Collins-Syndrome, shown on a family over three generations (author's transl)]. A Treacher Collins-syndrome has been followed up in a family over 3 generations. We found a change in deafness from grand parents- and parents-generation to the third one. The first two showed an inner ear deafness, the latter the well-known conduction deafness. The phenotype becomes more and more extensive in the following generations. Simultaneous with this development the deafness becomes more typical. In theory the inner ear deafness could either be explained as the final stage of a long standing conductive deafness or it is part of an incomplete developed syndrome.


[Objective tinnitus caused by arteriovenous malformations in the region of transverse and signoid sinus (author's transl)]. Surgical problems concerning the treatment of objective tinnitus caused by arteriovenous malformations of the posterior fossa are discussed. Angiographies and clinical signs of seven patients are reported. In two cases the operative procedure of Hugosson and Bergstroöm was sucessful. In principle the transverse and sigmoid sinus are exposed and separated from the arterial shunts.


[The difference between the noise deafness of the pitmans and the typical noise-induced hearing loss of the worker in metals (author's transl)]. Reffering to 92 expert evidences on the noise-deafness the signs are shown, in which the mining noise-deafness differs from the typical noise deafness in the metallurgical industry. In more than 80% of the cases a slope of the curve could be observed in the audiogram beginning at 125 cps. Therefore the hearing loss early impairs the ability to understand conversational speech. Only every second patient shows a positive result at the SISI-test; this is the reason, that the mining noise deafness frequently is misinterpreted. In these cases the cochlear hearing loss will be surely proved by the Langenbeck-audiogram and Békésy-audiogram. A secondary finding was the typical "mining ear-drum". These ear drums are bilaterally whitish, without reflex and thickened; they don't show any air-bone gap in the audiogram. The differences between the mining noise-deafness and the typical noise-induced hearing loss are explained by an additional baro-trauma which occurs, when the pitmans work at a depth of about 2000 feet.


[Evoked response audiometry of the guinea pig before and after drop in hearing induced by furosemid (author's transl)]. Using particular surface electrodes evoked response audiometry (ERA) can be applied to small laboratory animals without any sedation or anaesthesia. In addition to other methods we have studied in this way the influence of extreme doses of Furosemid on the guinea pig inner ear. 6-20 mg Furosemid per 100 g of bodyweight (about 100 times the human therapy dose) were applicated i.v. within 5 min resulting in a drop of hearing immediately. For the anaesthesized animal the cochlear microphonic potentials (CM) decrease simultaneously, indicating Furosemid to act at the inner ear itself. By both methods recovery of inner ear function is seen to start about 10 min later according to the little biological half life of Furosemid. As being restricted in respect of time for the CM-measurements we are sure about reversibility only by the ERA-method.


[Karyometric studies of the ventral cochlear nucleus in kanamycin treated guinea pigs (author's transl)]. The aim of these investigations was to study the effect of kanamycin on the nucleus ventralis cochleae in guinea pigs. The evaluation of the results was based on the karyometric method which is a very sensitive criterion of the cell activity. In the same animals, the microphonic potential (PM) and the nerve action potential (AP) were measured in order to detect changes in the peripheral auditory system. The results were statistically analysed. The studies showed that despite marked changes in the PM and AP values, there were no changes in the cell volume in the nucleus ventralis cochleae. The findings indicate that the injuries induced by kanamycin are localized peripherally and the later central morphological changes reported in the literature can be assumed to be secondary resulting from the damage to the peripheral auditory system by kanamycin.


[Modern methods of partial nasal reconstruction (author's transl)]. On the basis of selected examples on the possibilities of partial reconstruction of the nose with special regard to cosmetically advantageous reconstructive methods is reported. Our preference is given to the frontotemporal flap following Schmid, which is performed in several operation stages and varied in dependence upon the position and largeness of nasal defects. However, for older patients the nasolabial flap is recommended, because it is an one-stage operation technique.


[The free dermis fat transplantation as adjunct in the surgery of the parotid gland (author's transl)]. In order to overcome the short comings of a total parotidectomy in reference to appearance and the possible Frey syndrom, the author proposes the free dermis fat transplant directly after the extirpation of the gland. This diminishes the formation of serous fluid in the operated area. Long term observations showed no signs of the Frey syndrome. Further experimental research and clinincal observations will determine if the free fat transplantation is a workable method to prevent the Frey syndrom.


An unusual presentation of systemic organophosphate poisoning. A man presenting with an acute skin reaction to a pesticide was found to have severe systemic organophosphate poisoning.


Antibiotic-resistant mutants of Bacillus subtilis conditional for sporulation. Among spontaneously occurring antibiotic-resistant mutants of Bacillus subtilis 168 we have identified a sub-class that is conditionally sporulative. Mutants in this sub-class are resistant to antibiotic during vegetative growth but are sensitive during sporulation. Mutants conditionally-resistant to erythromycin, kanamycin, spectinomycin, and streptomycin have been isolated and characterized by phase contrast microscopy and with respect to their ability to synthesize heat-resistant endospores or the sporulation-associated enzyme alkaline phosphatase. The results suggest that several entirely different genetic lesions may result in this single phenotype. This group includes mutants whose properties suggest that both th 30S and 50S ribosomal subunits may be altered concomitant with early spore specific metabolism. The blockage imposed by antibiotic may be at or near Stage 2 of sporulation.


Effects of some antibiotics on the stringent control of RNA synthesis in Escherichia coli. Effects of neomycin, spectinomycin, tetracycline and chloramphenicol on the stringent control RNA synthesis and on ppGpp synthesis in the rel+-cells of Escherichia coli having a temperature-sensitive valyl-tRNA synthetase were examined. Without antibiotics, ppGpp began to accumulate and both RNA and protein syntheses were inhibited by transferring the exponentially growing cells from 30 degrees C (permissive temp.) to 40 degrees C (non-permissive temp.). Tetracycline or chloramphenicol, when added after the temperature shift, caused a resumption of RNA synthesis and decay of the accumulated ppGpp, while neomycin or spectinomycin had little effect both on RNA synthesis and the level of ppGpp. When the cells were treated with these antibiotics at permissive temperature, the shift of the temperature to 40 degrees C caused neither inhibition of RNA synthesis nor an accumulation of ppGpp. When neomycin or spectinomycin was added at the beginning of the temperature shift, RNA synthesis continued with an accumulation of ppGpp. Tetracycline or chloramphenicol had no such effect under the same conditions; RNA synthesis continued without an accumulation of ppGpp.


Mobilization of plasmid-borne drug resistance determinants for transfer from Pseudomonas aeruginosa to Escherichia coli. RSU2, a plasmid transmissible between strains of P. aeruginosa but not to Escherichia coli can be mobilized by R751. Conjugatants receive a single plasmid composed of DNA from both R751 and RSU2 which has the compatibility properties of a member of group P (like R751). Study of this fusion plasmid suggests that the failure of RSU2 to transfer into enterobacteria is due to an inability to replicate in these bacteria. The fusion plasmid replicates using the genes of R751.


[The mechanism of the acute action of insulin (author's transl)]. All the known acute actions of insulin are based on the reaction of insulin with specific insulin receptors in the cell membranes of the insulin-sensitive organs. The reaction of insulin with these receptors leads, through changes in the membrane structure, to changes in the permeability of the cell membrane, the activity of the normal membrane K+/Na+ATPase and adenyl cyclase. From this results the multiplicity of acute insulin effects on numerous metabolic quantities, following a kind of intensifier system. Under physiological conditions, the effect of insulin on the liver metabolism plays the central role for the maintenance of blood sugar homeostasis.


Evaluation of carotid stenosis by phonoangiography. Quantitative spectral (frequency) analyses were performed on 60 carotid bruits in 48 patients with suspected carotid stenosis. The technic was totally noninvasive. Bruits were recorded at the skin surface, analyzed by a minicomputer, and the degree of arterial stenosis estimated using a recently derived theory of sound production by turbulent blood flow. This method has been named phonoangiography. Fifty of the 60 bruits (83 per cent) had spectrums characteristic of turbulent blood flow, and a residual lumen diameter could be calculated. Results from the noninvasvie technic compared well with data obtained from radiographic carotid arteriograms in the 50 bruits with internal carotid artery stenosis. The residual lumen diameter estimated from phonoangiography differed from the radiographic value by less than 1 mm in 83 per cent and less than 1.5 mm in 92 per cent of the studies. Phonoangiography appears to be a useful method for assessment of carotid artery stenosis.


Fibroblasts in Huntington's disease.

Fibroblasts in Huntington's disease. We investigated the growth of skin fibroblasts in tissue culture from 10 patients with Huntington's disease and eight healthy, unrelated controls. The patients' ages ranged from 34 to 56 years (mean 48.5), and the mean duration of their clinical illness was 12.4 years. The controls' ages ranged from 32 to 64 years (mean, 42.3). No statistically significant differences were observed between the two groups in out-growth of cells from the biopsies nor during subsequent routine culturing of fibroblasts. This contradicts the finding of earlier investigators that skin fibroblasts from patients with Huntington's disease grow poorly in tissue culture. Matched pairs of Huntington's disease and control cultures grew at the same rate, but Huntington's disease cells grew to a significantly higher maximal density (P less than 0.02). This may indicate a genetically determined change in the cell surface or metabolic difference in Huntington's disease.


Effects of monovalent cations on cardiac  Na+, K+-ATPase activity and on contractile force. The relationship between Na+, K+-ATPase inhibition by monovalent cations and their inotropic effect was studied in guinea pig hearts. The activity of partially purified cardiac enzyme was assayed in the presence of 5.8 mM KC1 and either 20 or 150 mM NaCl. Rb+ and Tl+ inhibited Na+, K+-ATPase activity, the magnitude of the inhibition by these cations being greater in the assay media containing lower Na+ concentrations.  Tl+ produced a dose-dependent inhibition of Na+, K+-ATPase activity in the presence of 20 mM Na+ and 75 mM K+, a cationic condition  similar to that of intracellular fluid. Other monovalent cations such as K+, Cs+, NH4+, Na or Li+ produced essentially no effect on the Na+, K+-ATPase activity or slightly stimulated it. In left atrial strips stimulated with field electrodes and bathed in Krebs-Henseleit solution (5.8 mM K+ and 145 mM Na+), addition of Cs+ failed to alter the isometric contractile force significantly. NH4+ and K+ caused a transient positive inotropic effect which was partially blocked by propranolol. The positive inotropic response to K+ was followed by a negative inotropic response. Rb+ produced a sustained, dose-dependent inotropic response reaching a plateau at 1-2 min, whereas Tl+ produced a dose=dependent positive inotropic effect which developed slowly over a 30-min period. The positive inotropic effects produced by Rb+ and Tl+ were insensitive to propranolol pretreatment. Concentrations of Tl+ and cardiac glycosides which produce similar inotropic effects appear to cause the same degree of Na+-pump inhibition. The onset of the positive inotropic response to Rb+ or Tl+ was not dependent on the number of contractions which is in contrast to the cardiac glycoside-induced inotropic response. Substitution of 20 mM LiCl for an equimolar amount of NaCl in Krebs-Henseleit solution produced a significantly greater inotropic response than that observed when sucrose was substituted for NaCl. It appears that, among monovalent cations, only sodium pump inhibitors produce a sustained positive inotropic response.


Decreased glycolytic flux rate in the isolated perfused rat brain after pretreatment with 6-aminonicotinamide. Cerebral energy metabolism was studied in the isolated perfused rat brain after 6-aminonicotinamide (6-AN; 35 mg/kg i.p.) administered to the intact animals 7 hrs before perfusion was started. The metabolic alterations in the isolated rat brains were such as reported for rat and mouse brain in vivo: Inhibition of 6-phosphogluconate dehydrogenase was followed by an accumulation of 6-phosphogluconate, leading to a decreased activity of glucosephosphate isomerse. This was reflected by increased levels of glucose and glucose 6-phosphate and decreased levels of fructose 6-phosphate, pyruvate and lactate. Since the concentration of lactate in the perfusate of the isolated brain was also lowered, 6-AN must have reduced the glycolytic flux rate.


[The chromosomal syndromes (author's transl)]. Considering the results of modern cytogenetics in the area of chromosomal abberation, we can adopt without restriction two demands formulated by G. Koch (1971). First, further systematic research on mentally subnormal children should be carried out to clarify whether forms of chromosomal-genetic abnormality based on previously unknown structural alterations in one or several chromosomes can be discovered among those labelled "mentally subnormal". Second, the chromosomal-genetic forms of mental subnormality should be differentiated psychologically in close cooperation with child and adolescent psychologists and psychiatrists.


[Clinical aspects of chromosomal aberrations - problems of semiotics and nosography]. Despite intense clinical and cytogenetic research for more than 15 years we are far from knowing any definite relations between karyotype and phenotype. Both cytogeneticists and clinicians are working on the methodological improvements which are still necessary. In the methods of clinical recording there is still very great weakness of exact documentation of findings. Above all there is a lack of reproducible qualitative, and especially quantitative standards which constitute the basis of any practicable nosography. Some procedures for quantifying dysplastic facial features which have hardly been described so far (graphic statistics) are reported briefly. Also the summation of symptoms, a method currently used in the field of chromosomopathy syndromes, is subject to critical consideration. In this group of diseases a total list of symptoms of about 250 items can be obtained which includes multilocular minor stigmata, dysplasias, errors of differentiation and gross malformations of organs. An extraordinarily high degree of overlapping of symptoms is characteristic of these syndromes and makes accurate diagnosis difficult. However, for the trisomy-syndromes we succeeded in working out a diagnostic guideline by differentiating between an unspecified basic symptomatology concerning all trisomias and a pattern-forming additional symptomatology of each single syndrome. Thereby the diverse total symptomatology is reduced to the crucial and the recognition of patterns in daily practice is facilitated considerably. The comparatively specific additional symptomatologies of trisomy 13 -- 14 (Patau), trisomy 17 -- 18 (EDWARDS), and trisomy 21 (DOWN) are demonstrated in graphic views.


[Clinical psychological studies on adolescent and adult mongols (author's transl)]. A group of 20 mongol adolescents and adults with an average age of 19 years was strictly parallelized with a group of 20 brain-damaged adolescents and adults of other diagnostic groups with respect to age and intelligence level (average age 19 and average intelligence 50.65--52.90). The hypothesis of a possible deterioration of performance and personality was checked with the following battery of tests: Hamburg-Wechsler intelligence test for adults, the motoricity test according to Walther and the Rorschach experiment. The average difference in the sub-tests of HAWIE primarily emphasize the psycho-organic disturbance in visual-motor coordination, which is seen to a much greater extent in mongols than in non-mongols. The significant difference in completing pictures indicates an intensification of psycho-organicity. In agreement with the observations of WUNDERLICH, the psychomotoricity of the mongols is significantly slower and less adroit than the psychomotoricity of non-mongol oligophrenias. Non-significant differences in the psycho-organic sign (P%, F+%) in disfavor the mongols were seen in the Rorschach experiment. Factorial analysis distinguished the two groups very clearly. In the three most important factors, the mongol group showed stronger signs of psycho-organicity and weak mental performance associated with an intensified neuroticism factor compared with the brain-damage oligophrenia group of other diagnostic origin.


[Social behavior, musicality and visual perception in monogloid children (author's transl)]. Forty-nine mongoloid and 48 non-mongol test persons of equivalent age and intelligence were selected and studied with respect to social behavior, speech disorders (observation of behavior), musicality and visual perception. There were significant differences in favor of the mongols with respect to social adaption. Speech disorders of all kinds occurred significantly more frequently in mongol children; stuttering was significantly more frequent in the boys. The mongol group did significantly better in the musicality test; the difference in the rhythmical part was highly significant. The average differences in the capacity for visual discrimination of colors, geometrical forms and the spatial relationship of geometrical forms were not significant.


[On the mental structure of mongolism (author's transl)]. Forty-nine mongol and 48 non-mongol test persons of equivalent age and intelligence were selected and investigated with respect to intelligence components (S-I-T), motoricity (LOS), fine motoricity (KP), memorizing capacity (BA), working tempo (BS) and organization of the perceptual field (MZT). There were significant differences in S-I-T in favor of non-mongols in verbal tasks and in the LOS in favor of the mongoloids in rhythmic tasks. There was a tendency for the mongoloids to have poorer results for working tempo and fine motoricity. The differences in capacity to memorize and organization of perception were not significant.


[The mongol who becomes older (author's transl)]. The intensive concern of society, pedagogics and medicine for the child with disturbed development places Down's syndrome in the center of a new and gratifying interest. People from the most diverse disciplines are now concerned with mongoloid children, many simply following the trend of the times. Whoever has the care not only of the mongoloid child, however, but also of adolescents and adults, is very well aware that there are a series of processes in the physical, spiritual and mental development of the child through to adulthood in which an apparently dynamic characteristic is inherent. Alone and concurrently with several factors they produce impairment of family life with which the relatives are no longer able to deal. The question as to the motive for handing over the mongol concerned to an institution or home often cannot be answered. There are numerous reasons which may apply alone or in combination. As the mongol becomes older, his relatives are also becoming older giving rise to further difficulties. The present study shows that the furture of the mongol child, i.e. his puberty, his adolescence and adulthood must be planned carefully and principally require pedagogical, therapeutic, sociological and organizational consideration and measures. At present the future of the patients under our care does not seem by any means secure and ordered. The euphoria which is obligatory in many places is not justified.


[Investigations of erythrocyte metabolism in trisomy 21 (author's transl)]. Although the present studies were carried out in a large number of patients, it is not possible to say wether (and what kind of) enzymes are genetically fixed on chromosome 21. It is to be hoped that further studies on the question of enzyme localization, particularly of phosphofructokinase, will afford some clarification.


Eimeria falciformis (Eimer, 1870) in specific pathogen free and gnotobiotic mice. The endogenous and exogenous stages of a species of murine Eimeria were examined and compared with descriptions of other species isolated from Mus musculus. Developing stages of the parasite were found in the epithelial cells of the caecum and colon. Three asexual generations were observed, the third becoming apparent on the fourth day concurrently with gametogeny. The total length of the prepatent period was 5 days. Oocysts of the coccidian, identified as Eimeria falciformis, were administered to a group of germfree mice. Despite a dose level of 1 x 10(6), the coccidium was not established. Histological examination indicated the failure of the sporozoite to penetrate its host epithelium. The possible causes of this phenomenon were discussed.


[Role of photoperiodism and temperature in the induction of diapause in the egg phase of Aedes caspius caspius Pall. (Diptera, Culicidae)]. The influence of two photoperiods (12 and 20 hours of light per day) and different constant (25, 20, 15.5, 12.5 and 4 degrees) and fluctuating temperatures on the hatching of the larvae of Ae. caspius caspius from Bairam-Ali (Turkmenia) has been studied experimentally. The induction of winter egg diapause has been established to be under photoperiodic and temperature control. In eggs laid by short-day treated females the tendency to enter diapause is more expressed (55.9%) as compared with those laid by long-day exposed females (1.6%); in this case the eggs were kept at 12.5 degrees and in shortday regime. The low temperature 4 degrees increases only slightly the number of diapausing eggs (13.9%). The temperature which fluctuate during twenty-four hours being combined with short-day regime is more effective (the action of low temperature coincides with the scotophase). Even in the progeny of long-day exposed females the termorhythms 13--21, 8--20 and 4--20 degrees induce to enter diapause correspondigly 27.4, 22.8 and 59.9 per cent of the eggs.


[Experience in the control of the mosquito Culex pipiens molestus Forsk. In Leningrad]. Systematic control of mosquitoes (C. p. molestus) has begun in Leningrad since 1968 when undertaken inspections revealed mosquitoes and their aquatic stages in 21.8% of basements. All hatching sites of mosquitoes were registered and sanitary, hydrotechnical and destroying measures were carried out there. During the period of 1968 to 1973 the basements were cleaned, in 509 of them hydroisolation was made and many were dried. Anually chemical treatment of basements was undertaken and for 6 years the treated area amounted to 6.00000 m2. As a result, the percentage of flooded basements decreased from 25.8 to 7.1% and the hatching sites of mosquitoes--from 21.8% to 9.1%. The total population of mosquitoes decreased 43 fold.


Inhibition of ouabain-sensitive ATPase by the saliva of patients with cystic fibrosis of the pancreas. A study has been made of the effect of saliva from children with cystic fibrosis of the pancreas (CFP) on various components of the ATP hydrolyzing enzyme system. The ouabain-sensitive ATPase activity of erythrocyte membranes prepared from intact erythrocytes preincubated with CFP saliva was 35 +/- 4 nmol Pi/mg fry wt membrane suspension/hr, compared with 48 +/- 7 nmol Pi/mg dry wt membrane suspension/hr when the erythrocytes were preincubated with control saliva. A calcium-activated component of ATPase was decreased from 202 +/- 30 nmol Pi/mg dry wt membrane suspension/hr in erythrocytes preincubated with control saliva, to 151 +/- 17 nmol Pi/mg dry wt membrane suspension/hr when the incubation was carried out with CFP saliva. In a second series of experiments, ultrafiltered saliva was added directly to an ATPase assay. The saliva from children with CFP brought about a mean decrease in ouabain-sensitive ATPase of 16% compared with control saliva.


Gel electrophoretic studies on proteoglycans and collagen of abnormal human growth cartilage: proteoglycan abnormalities in pseudoachondroplasia and in Kniest's disease. The microchemical study of growth cartilage biopsies may improve the classification and the genetic advice of some types of growth disturbances and contribute to the understanding of biochemical defects. Small tibial growth cartilage biopsies were performed during orthopedic surgery in cases with achondroplasia, pseudoachondroplasis (three cases), Kniest's disease (two cases), diastrophic dwarfism (two cases), parastrematic dwarfism, pycnodysos tosis, mucolipidosis type III, Blount's disease, and in three normal growing children. Five human fetal cartilages were also studied. The proteoglycans were extracted with 4 M guanidinium chloride. After dialysis against 8 M urea at pH 7, the proteoglycans were obtained by ion chromatography in urea on DEAE-cellulose and submitted to gel electrophoresis on polyacrylamide-agarose gels. The gel electrophoresis of the proteoglycans of growth cartilage of normal growing children gave two metachromatic bands situated close one to another. The proteoglycans extracted from fetal growth cartilage gave a single band with a slightly slower migration. An abnormal gel electrophoretic pattern was found in pseudoachondroplasia and in Kniest's disease. In pseudoachondroplasia a single wide band was found; in overcharged tubes several thin, more rapid bands appeared in addition to the main band. In Kniest's disease three bands were found. In all of the other syndromes studied two normally or almost normally situated bands were present. Small differences in the width and intensity of the bands observed in several cases were difficult to assess. In all cases except mucolipidosis III and Kniest's disease the collagen was extracted by using limited cleavage and solubilization with pepsin, purified and analyzed by polyacrylamide gel electrophoresis. A type of collagen with a single alpha band was found.


Determination of diffusivity of oxygen and carbon dioxide in respiring tissue: results in rat skeletal muscle. Gas transfer rates for O2 and CO2 through freshly excised respiring rat abdominal muscle were measured. The tissue separated as a thin membrane two chambers, one of which was ventilated with a constant gas mixture. The other chamber was closed and the time course of changes of PO2 and PCO2, initially set at varied levels, was followed by electrodes. A plot of rate of change of PO2 and PCO2 in the closed chamber against the partial pressure difference across the tissue yielded both Krogh's diffusion constant, KO2 and KCO2, and metabolic rate of tissue, i.e. specific O2 consumption and CO2 production, mo2 and mco2. The mean values at 37 degrees C, KO2 = 1.31 x 10(-9) mMol-cm-1-min-1-torr-1 and KCO2 = 28.0 x 10(-9) mMol-cm-1-min-1-torr-1, did not differ significantly from values determined by other authors in various tissue preparations in which metabolism had been suppressed. Average O2 consumption, mo2 = 0.87 mMol-min-1-L-1, was not different from the values obtained in the same tissue by the Warbung manometric method, 0.74 mMol-min-1-L-1. The mean respiratory quotient, calculated as the ratio of mean CO2 production and mean O2 consumption, was 0.85.


The onset and progress of transformation of avian slow into fast muscles under neural influence. The slow anterior latissimus dorsi (ALD) muscles of newly hatched chickens were transposed and cross0innervated by the mixed, predominantly fast superior brachialis nerve, and investigated 2 to 15 months after the operation. Two months after the operation, myosin ATPase activity of the cross-innervated ALD muscles was still as low as in the control ALD, although the ultrastructure and the histochemical ATPase activity already showed a mixed fibre-type pattern with a predominance of fast -type fibres around the site of nerve implantation. The change of myosin properties of thw whole cross-innervated ALD did not occur until the third month after the operation. At that time, the myosin ATPase activity increased about 2.5 times and light chains of myosin of the fast type appeared in the electrophoretic pattern. The myosin ATPase activity attained 62% of the activity found in the control fast posterior latissimus dorsi muscles at three months; subsequently it remained at about this level reaching 68% 18 months after the operation. The results indicate that approximately two thirds of the cross-innervated ALD muscle fibres became changed towards the fast type under neural influence, whereas about one third remained slow, being re-innervated by the slow-type motor fibres of the implanted nerve.


Studies on the biological role of DNA methylation: inhibition of methylation and maturation of the bacteriophage phichi174 by nicotinamide. Nicotinamide was found to be a potent inhibitor of DNA methylation in vivo without interfering with protein or DNA synthesis. The inhibition of DNA methylation in a phage-infected cell resulted in a parallel decrease in the production of viable virus particles. In vitro experiments revealed that nicotinamide inhibits DNA methylase activity in a competitive fashion with respect to S-adenosylmethionine and non-competitively with respect to DNA. These results were interpreted to mean that DNA methylation is an essential step in the process of maturation of the bacteriophage phichi174.


Proteoglycans of soluble fraction of mouse mastocytoma. Proteoglycans have been isolated from a high speed supernatant fraction of a mouse mastocytoma by procedures which should minimize alteration of the native protein-polysaccharide molecule. The methods used include in vivo labeling proteoglycans with 35S-sulfate, 3H-leucine and 3H-lysine, centrifugation of the tumor homogenate at 105,000 g, cetylpyridinium fractionation of the supernatant, and further purification of some of the fractions obtained by DEAE-cellulose column chromatography, gel filtration on Sepharose 4B and cellulose acetate electrophoresis. Two major sulfated proteoglycans were obtained, one containing keratan sulfate-like material (KSP-S), the other a heparin-like polymer (HP-S). The presence in HP-S of a compound similar to heparin was confirmed by its digestibility with flavobacterium heparinase. HP-S contained about 4 per cent protein. Glycine was the predominant amino acid, and serine did not appear to be involved in the peptide-carbohydrate linkage. The proteoglycan present in HP-S appeared to be homogeneous when examined using cellulose acetate electrophoresis. KSP-S was found to contain sialic acid and its protein content was significantly higher than that of HP-S. Glutamic and aspartic acids were the most abundant amino acids in KSP-S.


DNA polyintercalating drugs: DNA binding of diacridine derivatives. As a first step in the synthesis and the study of DNA polyintercalating drugs, dimers of acridines have been prepared. Their DNA binding properties have been studied. It has been determined that when the chain separating the two aromatic rings is longer than a criticål distance, bisintercalation is actually observed and that the DNA binding affinity becomes quite large (greater than 10(8)-10(9) M-1). It is shown also that the optical characteristics of these molecules are dependent on the sequences of DNA. The fluorescence intensity of one of these dimers when bound to DNA varies as the fourth power of its A+T content. This derivative could be used as a fluorescent probe of DNA sequence.


Arrangement of sequences in the inverted terminal repetition of adenovirus 18 DNA. In contrast to the single-stranded circular molecules produced with denatured DNA from other adenoviruses, there was associated with nearly all circular molecules of adenovirus type 18 a visible, duplex projection. These projections had a mean contour length of 0.31 +/- 0.12 mum, equivalent to approximately 3% of genome length. Individual projections ranged in size from 0.1 to 2 mum. Alkaline sucrose gradient purification of single-stranded molecules did not affect formation of these projections, and treatment of a preparation of circular molecules with Neurospora crassa single-strand specific nucleases yielded 0.34 +/- 0.09 mum duplex fragments. Single-stranded circles did not form if a limited number of nucleotides were removed from the 3' ends of native molecules by Escherichia coli exonuclease III digestion prior to denaturation and annealing. In addition, preformed single-stranded circles could be converted to linear molecules by similar treatment. Based on the formation of specific heteroduplex structures when preparations of native DNA were denatured and reannealed and the absence of branches on linear, single-stranded molecules, we conclude that projections are generated by unusually long, inverted terminal repetitions. The repetitious sequences occur in place of rather than in addition to regular sequences. These data provide direct, visual evidence for the arrangement of bases in the inverted terminal repetition of adenovirus DNA.


Electron microscope study of native and crosslinked rabbit muscle phosphofructokinase. Electron microscopy of native and dimethyl-suberimidate crosslinked rabbit muscle phosphofructokinase (EC 2.7.1.11; ATP:D-fructose-6-phosphate 1-phosphotransferase) has been carried out using negative staining with sodium phosphotungstate. The results obtained suggest the protomer of molecular weight 80,000 can be approximated as a prolate ellipsoid with axes of 67 A and 25 A. The dimer, which is the fundamental unit for polymerization, is formed by association along the 25 A axis and has approximately dimensions of 67 A X 55 A X 25 A. The tetramer appears to be formed by an end-to-end aggregation of dimers, and the octamer is a sheet-like structure made up of a side-to-side aggregation of tetramers. Higher crosslinked aggregates and long crosslinked filaments also are seen. The filaments have a constant width of about 250 A, are about 0.5 mum in length, and appear to involve tetramers as a fundamental structural unit. The functional significance of the filament structure is not known.


Association of phiX174 DNA-dependent ATPase activity with an Escherichia coli protein, replication factor Y, required for in vitro synthesis of phiX174 DNA. phiX174 DNA-dependent DNA synthesis is catalyzed in vitro by the combination of at least 11 purified protein fractions: dnaB, dnaC(D), and dnaG gene products, DNA polymerase III, DNA elongation factors I and II, DNA binding protein, and replication factors W, X, Y, and Z. The reaction requires ATP, 4 dNTPs, and Mg+2 and is specific for phiX174 (or phiXahb) DNA. Purified replication factor Y contains phiX174 (or phiXahb) DNA-dependent ATPase (or dATPase) activity. The ATPase activity is poorly stimulated by other single-stranded DNA, by double-stranded DNA, or by RNA. The products of the phiX174 DNA-dependent ATPase activity of factor Y are Pi and ADP (or dADP). The association of phiX174 DNA-dependent ATPase activity with factor Y was shown in the following ways: (a) the two activities copurified with a constant ratio; (b) they comigrated on native polyacrylamide gel electrophoresis; (c) both activities were heat-inactivated at the same rate; and (d) both showed identical patterns of N-ethylmaleimide sensitivity.


Carbon-13 nuclear magnetic resonance study of chondroitin 4-sulfate in the proteoglycan of bovine nasal cartilage. The Fourier transform 13C nuclear magnetic resonance spectra of bovine nasal cartilage proteoglycan subunit and complex and whole bovine nasal cartilage were obtained and compared with that of chondroitin 4-sulfate. The spectrum of chondroitin 4-sulfate in solution revealed multiple resolvable resonances with linewidths that are consistent with considerable segmental motion in the polysaccharide chain. The spectra of proteoglycan subunit and complex in solution and that of whole cartilage were very similar to that of free chondroitin 4-sulfate chains. This indicates that the linkage of multiple chondroitin sulfate chains to proteoglycan core protein and the association of proteoglycan with collagen and other constituents of cartilage matrix does not significantly alter the structure and motions of these chains.


Antigenic and enzymatic architecture of Micrococcus lysodeikticus membranes established by crossed immunoelectrophoresis. By crossed immunoelectrophoresis with membrane antiserum, 17 antigens have been detected in fractions from plasma membranes of M. lysodeikticus solubilized with Triton X-100. Absorption tests with protoplasts have demonstrated that eight of the antigens are expressed on the surface. Of these antigens the major one has been identified as a succinylated mannan. Five of the principal immunoprecipitates unaffected by absorption with protoplasts were shown by zymograms to possess the following enzymic activites: succinate dehydrogenase (EC 1.3.99.1), ATPase (EC 3.6.1.3), NADH dehyrogenase (EC 1.6.99.3)(two separate components), and malate dehydrogenase (EC 1.1.1.37). These enzymes or enzyme-complexes are, therefore, not expressed on the outer surface of the protoplast membrane.