The mechanism of increase in the ATPase activity of sarcoplasmic reticulum vesicles treated with n-alcohols. Treatment of sarcoplasmic reticulum vesicles with aqueous n-alcohols caused inhibition of calcium uptake and enhancement of ATPase activity. With increasing alcohol concentration, the ATPase activity reached a maximum (in the case of n-butanol, at about 350 mM) and then decreased. The effect of n-butanol was extensively studied. The purified ATPase enzyme and leaky vesicles treated with Triton X-100 or phospholipase A showed high ATPase activity in the absence of n-butanol. With increasing n-butanol concentration, their atpase activities began to decrease above about 250 mM n-butanol, without any enhancement. In the presence of ATP, the turnover rate of calcium after calcium accumulation had reached a steady level was the same as that at the initial uptake. n-Butanol did not affect these rates. Kinetic analyses of these experiments were carried out. The mechanisms of calcium transport and of increase of ATPase activity in the presence of alcohol were interpreted as follows. After calcium accumulation had reached a steady level, fast influx and efflux continued; the influx was coupled with phosphorylated enzyme (E-P) formation and most of the efflux was coupled with rephosphorylation of ATP from ADP and E-P. The observed ATPase activity is the difference between these two reactions. If alcohol molecules make the vesicles leaky, calcium ions will flow out without ATP synthesis and the apparent ATPase activity will increase. The effect of alcohols on sarcoplasmic reticulum vesicles was separated into two actions. The enhancement of ATPase activity was attributed to a leakage of calcium ions from the vesicles, while the decrease of ATPase activity at higher concentrations of alcohols was attributed to denaturation of the ATPase enzyme itself. The two effects were interpreted in terms of equilibrium binding of alcohol molecules to two different sites of the vesicles; leakage and denaturation sites. Similar analysis was carried out for various n-alcohols from methanol to n-heptanol. The apparent free energies of binding of the methylene groups of n-alcohols were evaluated to be -863 cal/mol for the leakage site, and -732 cal/mol for the denaturation site.


Inhibition of proteases in coagulation, kinin-forming and complement systems by alpha2-plasmin inhibitor. Inhibitory activities of alpha2-plasmin inhibitor against various proteases were investigated. The inhibitor promptly inhibited the esterolytic activity of alpha-chymotrypsin and progressively inhibited the esterolytic or amidolytic activities of bovine plasma kallikrein, bovine thrombin and bovine activated factor X. Heparin had no effect on the reaction of the inhibitor with thrombin or activated factor X. However, the inhibitor had no effect on the activities of human C-1-esterase, papain and snake venom kininogenase. On the basis of its rapid inhibition of kallikrein, alpha2-plasmin inhibitor is considered to exert some regulating effect on kallikrein activity in plasma.


Acanthamoeba cofactor protein is a heavy chain kinase required for actin activation of the Mg2+-ATPase activity of Acanthamoeba myosin I. We have purified a cofactor protein previously shown (Pollard, T. D., and Korn, E. D. (1973) J. Biol. Chem. 248, 4691-4697) to be required for actin activation of the Mg2+-ATPase activity of Acanthamoeba myosin I. The purified cofactor protein is a novel myosin kinase that phosphorylates the single heavy chain, but neither of the two light chains, of Acanthamoeba myosin I. Phosphorylation of Acanthamoeba myosin I by the purified cofactor protein requires ATP and Mg2+ but is Ca2+-independent. The Mg2+-ATPase activity of phosphorylated Acanthamoeba myosin I is highly activated by F-actin in the absence of cofactor protein. Actin-activated Mg2+-ATPase activity is lost when phosphorylated Acanthamoeba myosin I is dephosphorylated by platelet phosphatase. Phosphorylation and dephosphorylation have no effect on the (K+,EDTA)-ATPase and Ca2+-ATPase activities of Acanthamoeba myosin I. These results show that cofactor protein is an Acanthamoeba myosin I heavy chain kinase and that phosphorylation of the heavy chain of this myosin is required for actin activation of its Mg2+-ATPase activity.


Aurovertin binds to the beta subunit of yeast mitochondrial ATPase. Isolated beta subunit of ATPase (F1) from yeast mitochondria does not catalyze an ATPase reaction but still binds the specific F1 inhibitor aurovertin. Binding was measured by enhancement of aurovertin fluorescence; it was as tight as that to F1-ATPase. No binding was observed with F1 or with isolated beta subunit from a single-gene nuclear yeast mutant whose F1-ATPase was resistant to aurovertin.


Modification of (Na+,K+)-ATPase function by purified antibodies to the holoenzyme. Effects on enzyme activity and (3H)ouabain binding. Antibodies (abys) raised to (Na+,K+)-ATPase were purified by elution methods and shown to be cross-reactive with anti-gamma-globulin and the original antigen. Abys were isolated from two different antisera and the effects on (Na+,K+)-ATPase hydrolytic activity and [3H]ouabain binding were measured. The antisera fractions differed as to their maximum level of inhibition of hydrolytic activity and maximal [3H]ouabain binding, but both fractions caused inhibition of maximal [3H]ouabain binding to the same quantitative extent as inhibition of hydrolytic activity. Variable effects on the rate of [3H]ouabain binding were noted which were highly dependent on ligand conditions. During the "turnover state conditions" of the enzyme, the abys stimulated the rate of [3H]ouabain binding to the (Na+,K+)-ATPase. We conclude that effects of aby-(Na+,K+)-ATPase interaction depend upon degree of purity of aby, specificity, aby/enzyme ratios, and ligand conditions.


Crawling patterns in neglected poliomyelitis in the Solomon Islands. Two hundred and twenty-two adult crawling poliomyelitic cripples were investigated. Analysis showed that bilateral lower limb paralysis with or without trunk involvement was the most common residual lesion. Remarkably paralysis was confined to a single lower limb in six cases. The various crawling patterns can be classified into six broad groups which are described and illustrated. It is suggested that in a developing country this classification has a practical application in the assessment of patients for rehabilitation to crutch walking. Subjects in the "true quandruped progression" and "squatting gait" categories generally have an excellent potential for rehabilitation, as often do those in the "infant-like crawl" group. Caution should be exercised in attempting crutch walking for those in the "body-dragging" group, and there seems little indication for attempting to establish upright walking for those in the "buttock pivoting" and "minimal movement" groups.


Ultrastructure of the sodium pump. Comparison of thin sectioning, negative staining, and freeze-fracture of purified, membrane-bound (Na+,K+)-ATPase. Purified (Na+, K+)-ATPase was studied by electron microscopy after thin sectioning, negative staining, and freeze-fracturing, particular emphasis being paid to the dimensions and frequencies of substructures in the membranes. Ultrathin sections show exclusively flat or cup-shaped membrane fragments which are triple-layered along much of their length and have diameters of 0.1-0.6 mum. Negative staining revealed a distinct substructure of particles with diameters between 30 and 50 A and with a frequency of 12,500 +/- 2,400 (SD) per mum(2). Comparisons with sizes of the protein components suggest that each surface particle contains as its major component one large catalytic chain with mol wt close to 100,000 and that two surface particles unite to form the unit of (Na+,K+)-ATPase which binds one molecule of ATP or ouabain. The further observations that the surface particles protrude from the membrane surface and are observed on both membrane surfaces in different patterns and degrees of clustering suggest that protein units span the membrane and are capable of lateral mobility. Freeze-fracturing shows intramembranous particles with diameters of 90-110 A and distributed on both concave and convex fracture faces with a frequency of 3,410 +/- 370 per mum(2) and 390 +/- 170 per mum(2), respectively. The larger diameters and three to fourfold smaller frequency of the intramembranous particles as compared to the surface particles seen after negative staining may reflect technical differences between methods, but it is more likely that the intramembranous particle is an oliogomer composed of two or even more of the protein units which form the surface particles.


On the distribution of Na+-pump sites in the frog skin. Exposure of the outside of the isolated frog skin to a Ringer's solution, made hypertonic by the addition of mannitol, causes a rapid and sustained increase in transepithelial permeability through a structural distortion-a focal blistering-of the "tight" junctions of the outermost living cell layer. [(3)H]ouabain, used as an autoradiographic marker for the Na+-pump (Na+-K+-adenosine triphosphatase), is usually unable to penetrate the frog skin from the outside solution, but when added to a hypertonic mannitol- Ringer's solution in the outside bath it readily penetrates the epithelium, presumably through the opened shunt pathway. Radioautographic analysis of [(3)H]ouabain binding sites revealed that most of ouabain enters from the outside solution binds to the sites on the cell membranes of the stratum spinosum, as was the case when it was applied from the inside bath in an earlier study. The outer living cell layer, the first to be exposed to ouabain, does not appear to be the major site for the Na+-pump, and therefore, is not likely to be responsible for most of the active pumping of Na+. This result demonstrates that previous failure to show a high density of Na+-pump sites on the cells of the outermost layer, when [(3)H]ouabain was applied from the inside solution, was not due to the inability of the marker to reach these cells at a sufficient concentration to reveal all pump sites. These results provide further support for a model of Na+-transport across the frog skin which distributes the active pump step on the inward facing membranes of all living cells.


Application of the somatization factor of the MMPI-168 with low back pain patients. The usefulness of the Somatization factor of the MMPI-168 with low back pain patients was examined in two separate studies. In study 1, 58 male veteran low back pain patients who had been divided into organic and mixed groups served as Ss. MMPI protocols were rescored for the five factors of Overall, Hunter, and Butcher (1973). The organic and mixed groups differed only on the Somatization factor. A cutting score of raw greater than or equal to 8 (T = 75) was determined to classify the sample correctly 74.5% of the time. In study 2, a second sample of 48 male veteran low back pain patients was divided into mixed, organic, and functional groups. The mixed group was subdivided further into a mixed-pain group who still were having pain and were seeking treatment and a mixed-relief group who were experiencing a reduction of pain and were returning to vocational activities. The functional and mixed-pain groups differed from the organic and mixed-relief groups on the Somatization and Depression factors. The cutting score determined in the study 1 correctly classified patients 83% of the time.


Study of the noncollagenous components of the periodontium. Periodontal ligament and gingivae of bovine and porcine periodontium were analyzed for relative amounts of carbohydrates, collagen, and acid mucopolysaccharides. The sugar content was 3.6% and 3.4% of dry weight in bovine and porcine periodontal ligament, respectively. The values were lower in the gingivae being 2.34% and 2.30%, respectively. Approximately 50% of hexosamine in gingivae was present in acid mucopolysacchrides as compared to 36% in periodontal ligament. Relative to collagen there was a considerable amount of non-collagenous glycoproteins present in the periodontium as judged by carbohydrate content of the tissue.


Muscle spindle supply to the human jaw muscle. Histological study of the human jaw muscle revealed that the temporal muscle displayed 342 muscle spindles, 208 in the horizontal and 134 in the vertical portion; the masseter muscle contained 114 spindles, 91 in the superficial and 23 in the profound portion; the medial pterygoid muscle had 59; and the lateral pterygoid muscle contained 6, four in the upper head and two in the lower head. These data suggest that the extensive mobility of the temporomandibular joint, and the maintenance of mandibular posture during mastication and speech are strongly influenced by proprioceptive mechanisms.


Circulating immune complexes and activation of the complement sequence in acute allergic bronchopulmonary aspergillosis. A 13-yr-old asthmatic girl with allergic bronchopulmonary aspergillosis (ABPA) exhibited eosinophilia, IgE hypergammaglobulinemia, serum precipitins to Aspergillus, a dual skin reaction to Aspergillus fumigatus, and positive, lymphocyte transformation in vitro in response to stimulation with Aspergillus fumigatus antigen. The acute phase of the disease was characterized by an increase in blood eosinophilia, a rise in serum IgE, circulating immune complexes, and evidence of activation of the complement system through the classical pathway.


Metabolism of crufomate [(4-tert-butyl-2-chlorophenyl methyl methylphosphormidate)] by the rat. Fifteen metabolites of crufomate (4-tert-butyl-2-chlorophenyl methyl methylphosphoramidate, I) were identified in the excreta from rats given single oral doses of I. Compound I was not detected in either the urine or the feces. The metabolic reactions observed were N-and O-demethylation, oxidations of the t-butyl moiety, replacement of the H-N-CH3 with an OH moiety, hydrolysis of the phosphoramidate moiety to yield the phenol, conjugation with glucuronic acid, and combinations of these reactions. No ring dehalogenation or ring substitution was observed.


Rat intestinal metabolism of crufomate (4-tert-butyl-2-chlorophenyl methyl methylphosphoramidate). Everted sacs of rat small intestine metabolized crufomate (4-tert-butyl-2-chlorophenyl methyl methylphosphoramidate) under in vitro conditions to form six 14C-labeled metabolites in quantities sufficient for isolation and identification. These metabolites were 4-tert-butyl-2-chlorophenyl methyl phosphoramidate (25%), 2-chloro-4(2-hydroxy-1,1-dimethylethyl)phenyl methyl methylphosphoramidate (19%), 2-[3-chloro-4-[[(methoxy) (methyl-amino)phosphoinyl]oxy]phenyl]-2-methylpropionic acid (2%), 4-tert-butyl-2-chlorophenol (0.8%) and its glucuronide (6%), and the aromatic glucuronide of 2-chloro-4(2-hydroxy-1,1-dimethylethyl)phenol (1%). These intestinal metabolites may represent precursory stages in the overall metabolism of crufomate.


Uranaffin reaction: a new cytochemical technique for the localization of adenine nucleotides in organelles storing biogenic amines. Amine storage organelles of aldehyde-fixed rabbit platelets have a strong affinity for uranyl ions before their dehydration and appear highly electron-dense when examined by electron microscopy; both the matrix and membrane of these organelles are intensely stained. This affinity, which is also shown by platelets of other species, including healthy reserpinized platelets, which contain no cytochemically demonstrable amine, show a positive uranaffin reaction. However, platelets and megakaryocytes of strains with storage pool deficiency (low adenosine-5'-triphosphate), including patients with Hermansky-Pudlak syndrome, are uranaffin negative. The cytochemical reaction, probably the result of an interaction between UO2++ ions and phosphate groups of 5'-phosphonucleotides, is also observed in adrenal medulla, sympathetic nerve terminals and ganglion cells, suggesting that the technique will be of considerable value in identification of aminergic neurons and in further elucidation of amine storage mechanisms.


Androgenic control of the harderian gland in the male golden hamster. In the golden hamster, there are marked sex differences in the Harderian gland. Male glands (which are heavier than female glands) possess two cell forms (Type I and Type II cells); female glands only exhibit the former. Female (but not male) glands contain large amounts of porphyrin, which are readily visible as solid depositions within the lumina. The weight, histology and porphyrin content of the Harderian gland was examined in intact adult male hamsters and in male hamsters castrated for 1,2 or 8 months. Castration resulted in a significant reduction in the weight of the gland, the disappearance of Type II cells, and the presence in the gland of solid porphyrin accretions. The levels of copro- and (especially) protoporphyrin were greatly increased. These changes were more marked with time after castration. When the ability of diverse androgens (testosterone, 5alpha-dihydrotestosterone, androst-4-ene-3,17-dione (androstenedione), dehydroepiandrosterone and androsterone) to prevent these changes was tested, testosterone and androstenedione maintained glandular weight. All the androgens maintained normal frequencies of Type II cells and all except dehydroepiandrosterone prevented deposition of porphyrin. The potencies of the various androgens in maintaining normal Harderian gland morphology and activity are compared with their effects on other somatic variables and sexual behaviour.


Regulatory mechanisms in cell-mediated immune responses. VI. Interaction of H-2 and non-H-2 genes in elaboration of mixed leukocyte reaction suppressor factor. Previous studies have shown that alloantigen-activated spleen T cells produce a soluble factor which suppresses mixed lymphocyte reaction proliferative responses, and that the interaction between suppressor and responder cells is controlled by genes of the H-2 complex. However a defect in the expression of suppressor activity was identified in the mouse strain C57BL/6J. Factor prepared from alloactivated B6 spleen cells failed to suppress MLR responses of syngeneic or H-2 compatible responder cells. Unimpaired suppressor factor production by other H-2 (b) strains and failure of suppressor factor production by a B6 congenic strain, B6.C-H-2(d) isolated the defective gene to the non-H-2 portion of the genome. In addition, the defect appeared to be related specifically to inability to produce an active factor, while the capacity to respond to suppressor molecules was unimpaired. The genetic character of the non-H-2 gene action was identified in F1 hybrid studies. Initially F(1) hybrids of the nondefective histoincompatible strains were studied. Suppressor factor from F1 cells suppressed the responses of both parental strains, and parental factors each suppressed the response of F(1) cells. Adsorption of F(1) factor with Con A-activated thymocytes of either parental strain removed suppressor activity specific for that strain, leaving activity against the other parental strain intact. The data support cedominant expression and production of distinct, parental H-2 haplotype-specific suppressor molecules by F(1) suppressor cells. An F(1) hybrid of the defective B6 strain with nondefective BALB/c produced suppressor factor which was also capable of suppressing both parental strains. Production of a suppressive B6-reactive factor by F(1) cells was verified by adsorption studies. Thus it appears that non-H-2 genes of the BALB/c parent acted in a genetically dominant fashion to provide the function required for expression of B6 suppressor molecules. We conclude that multiple genes control the expression of alloactivated suppressor cell activity, with at least one gene mapped to the I-C subregion of the murine major histocompatibility complex and one or more genes mapped to the non-H-2 gene complement.


Quantitative studies on the precursors of cytotoxic lymphocytes. III. The lineage of memory cells. In the course of a mixed lymphocyte culture, memory cells are produced which can give rise to a large secondary cytotoxic lymphocyte response on reexposure to the sensitizing alloantigen. We have studied the lineage of these memory cells using a clonal assay for precursors of cytotoxic T lymphocytes (CLP). Our data provide conclusive evidence that individual CLP, upon stimulation with alloantigens, gives rise to clones which contain memory cells of the same specificity as the CLP. Only half of the clones that responded in the primary stimulation could be reactivated upon exposure to the original priming alloantigen.


H-2 effects on cell-cell interactions in the response to single non-H-2 alloantigens. II. H-2D region control of H-7.1-specific stimulator function in mixed lymphocyte culture and susceptibility to lysis by H-7.1-specific cytotoxic cells. The relative immunogenicity of the H-7.1 alloantigen has been shown in a previous communication to be regulated by a gene in the D region of the mouse major histocompatibility (H-2) complex. The level of relative immunogenicity was inferred from survival times of H-7.1-incompatible skin grafts donated by donors with different H-2 haplotype origins of H-2D region genes. In this communication we report the results of an extension of these previous investigations into the possible role of H-2D region genes in controlling the capacity of H-7.1-incompatible lymphocytes to stimulate H-7.1-speciflc mixed lymphocyte culture proliferation and generation of cytotoxic effector cells. The results reported herein demonstrate that the H-2D genotype of H-7.1-incompatible stimulator cells determines the relative H-7.1-specific capacity of those lymphocytes to stimulate H-7.1-specific proliferation of in vivo primed responder T cells in secondary mixed lymphocyte culture. H-2D(b)-bearing, H-7.l-incompatible stimulators were significantly more effective in stimulating H-7.1-specific proliferation than H-2D(d)-bearing stimulators. As expected, H-2D(b), H-7.1-in-compatible stimulators were also more effective than H-2D(d) a stimulators in generating H-7.1- specific cytotoxic effector cells. Further, the susceptibility of (51)Cr- labeled, H-7.1-incompatible lymphoblast targets to H-7.1-specific lysis was similarly regulated by an H-2D gene. Reciprocal H-2 restriction (F(1) cells are capable of killing only the cells bearing the immunizing cell parental H-2 haplotype) observed by other investigators for cytolysis of non-H-2-incompatible targets was not observed. H-2D a-bearing, H-7.1- incompatible stimulators stimulated generation of cytotoxic effectors capable of detectably lysing H-2D(b) but not H-2D(a)-bearing, H-7.1- incompatible targets. The impact of these observations on the proposed models for H-2 restriction of non-H-2 histocompatibility antigen-specific cytolysis is discussed.


Occurrence of restricted suppressor T-cell activity in man. Blood leukocytes from an immunologically hyporesponsive patient with urinary bladder carcinoma were found to be deficient in their ability to stimulate 3 of 27 responder leukocyte preparations from normal individuals in one-way mixed leukocyte culture (MLC). The patient's T-depleted leukocytes, however, functioned adequately as stimulator cells. T-enriched lymphocytes from this patient suppressed the MLC responsiveness of those three normals but not the responsiveness of other normals. The patient's cells suppressed the MLC responsiveness of only one of each of the parents of two of the normals who could be suppressed by the patient's leukocytes suggesting a possible genetic restriction to this suppressor cell activity.


The role of gene products of the I-J subregion in mixed lymphocyte reactions. We have examined the MLR reaction in two sets of recombinants that differ in the I-J subregion. In both cases, significant stimulation was mediated by antigens controlled by genes in the I-J subregion. This stimulation was inhibitable by the addition of the culture of antisera directed against the I-J gene products on the stimulator cell. The specificity of this inhibition was shown by specific blocking of the relevant gene product on F1 hybrid stimulator cells. MLR stimulation was also eliminated by pretreatment of the stimulator population with anti-I-J sera plus complement. Pretreatment of F1 hybrid stimulator T cells with anti-I-J sera directed against either parental I-J product in the presence of complement, completely eliminated stimulation, indicating that there is no allelic exclusion of the relevant I-J products. Pretreatment with an anti-I-E/I-C serum and complement also eliminated stimulation, suggesting that the stimulating T cells express both I-J and I-E/I-C subregion products. This assay offers a potentially more direct and practical method for serological detection of the I-J products.


Autologous mixed lymphocyte culture reactions and generation of cytotoxic T cells. Autologous mixed lymphocyte culture (MLC) reactions were studied utilizing autologous purified B cells and autologous established B lymphoid cell lines as stimulating cells. Similar results were obtained although somewhat greater stimulation of lymphocyte proliferation was found with the autologous lymphoid cell lines. Cytotoxic T cells were not generated against the stimulating cells in either case when peripheral blood cells were used as targets. A low cytotoxicity was detected when lymphoid cell lines were used both as stimulators and target cells. However this was nonspecific and was always greater for heterologous lines than for the stimulator line. Third-party cell experiments demonstrated that the autologous reaction could serve as a proliferative stimulus for specific cytotoxic lymphocyte generation. Heat-treated allogeneic lymphocytes that alone do not stimulate proliferation ro cytotoxic T-cell generation in MLC reactions when added to the autologous system produced specific cytotoxic cells. The separation of the proliferative phase from the cytotoxic cell generation was especially striking in these experiments. Possible uses of this system for the generation of specific cytotoxic cells to other nonstimulatory cells are discussed.


Lymphocyte transformation induced by autologous cells. V. Generation of immunologic memory and specificity during the autologous mixed lymphocyte reaction. Lymphocyte proliferation in vitro may follow antigen recognition and serve as a correlate of cell-mediated immunity. Lymphocyte proliferation can also be simulated by nonimmune mechanisms as, for example, following culture with plant lectin, lipopolysaccharides, or staphylococcal protein A (1). The autologous mixed lymphocyte reaction (MLR) refers to the proliferation of T lymphocytes cultured with autologous mon-T lymphocytes (2,3). The purpose of this study was to determine whether lymphocyte proliferation in the autologous MLR results from immune or nonimmune mechanisms. We have shown that the autologous MLR has two classical attributes of an immune phenomenon: memory and specificity.


Acne vulgaris: recent advances in pathogenesis and treatment. Hormonal factors, particularly androgens, appear to be important in the pathogenesis of acne vulgaris. The sebaceous glands in acne are more sensitive to normal blood levels of androgens, and are stimulated to produce more sebum. Corynebacterium acnes in the sebaceous follicles act on triglycerides in the sebum to form free fatty acids which might alter the process of keratinization in the follicular canal. A microcomedo is formed which can progress to the clinical lesions of acne. Sebum and its components may also be inflammatory if released into the skin. There are, however, still a number of unanswered questions relating to acne pathogenesis. Currently, therapy of acne vulgaris revolves around topical benzoyl peroxide and retinoic acid and systemic tetracyclines. Benzoyl peroxide and tetracyclines are antibacterial while retinoic acid is comedolytic. Because of these different actions, combined therapy appears to be more effective (benzoyl peroxide and/or tetracyclines together with retinoic acid). Topical antibiotics show promise as new therapeutic agents.


Rapid modulation of gill Na+ + K+-dependent ATPase activity during acclimation of the killifish Fundulus heteroclitus to salinity change. The enzymatic properties of membrane-bound Na+ + K+-ATPase from gills of killifish acclimated to fresh water, to 16% sea water, or to 30% sea water appear to be identical, indicating that the same enzyme may function to absorb Na+ in low salinities and excrete Na+ at the gills in high salinities. Ammonium ion is an effective substitute for K+: in the ATPase reaction itself, in blocking phosphorylation of the ATPase protein, and in inhibiting the binding of ouabain to the enzyme. The specific activities of the Na+ + K+-ATPase in the three different salinities are consistent with the expected Na+ pumping rates: higher in fresh water and 30% sea water than in 16% sea water. Within one-half hour after transfer of killifish from one salinity to another, gill Na+ + K+-ATPase activities reach equilibrium levels. The rapid increase in Na+ + K+-ATPase activity in gill microsomes of fish acclimating from fresh water to 30% sea water is accompanied by a slow decrease in the number of binding sites for ouabain, supporting the idea that acclimation to short-term salinity changes may involve modifications in the catalytic rate rather than the number of Na+ + K+-ATPase molecules.


Pathways of glucose metabolism in Candida 107, a lipid-accumulating yeast. Phosphofructokinase was not detected in extracts of Candida 107 prepared in a variety of ways but was highly active in cells treated with toluene. Disruption of these cells destroyed activity of phosphofructokinase indicating that the enzyme is extremely labile. As patterns of labelling from [I-14C]glucose and [6-14C]glucose showed that 60% of glucose was metabolized via the pentose cycle, augmentation of this cycle is necessary to account for the high molar growth yields of this yeast. Phosphoketolases, reacting with xylulose 5-phosphate and fructose 6-phosphate, were found but the extent to which they contribute to glucose metabolism was not assessed.


Ouabain binding sites in skeletal muscle from normal and dystrophic mice. The specific binding of tritiated ouabain was used to estimate the density of Na+-K+-ATPase sites ("Na+-pump" sites) in segments of skeletal muscle from normal and dystrophic mice. Ouabain binding was approximately 4 times greater in red (soleus) muscle than in white (superficial gastrocnemius) muscle from normal animals. In dystrophic soleus muscles, ouabain binding was decreased by nearly one-half. Because Na+-K+-ATPase activity is associated with plasma membranes, these observations constitute further evidence for a sarcolemmal abnormality in dystrophic mice.


Neurotransmitter enzyme abnormalities in senile dementia. Choline acetyltransferase and glutamic acid decarboxylase activities in necropsy brain tissue. Reductions in 2 neurotransmitter synthesizing enzymes in brain, glutamic acid decarboxylase (GAD) and choline acetyltransferase (CAT), have been found in dementias of different origins, including senile dementia (Alzheimer type). Significant reductions in cerebral GAD have also been found in depression (unipolar). The GAD reductions did not generally appear to be localised in any specific region of the brain examined. However, the reduction of CAT in the hippocampus, relative to reductions in other areas examined, was substantially greater in the brains with Alzheimer-type changes. GAD and CAT activities in normal brains were examined for the effects of some variable factors inherent in necropsy biochemical measurements. These factors included: (i) age; (ii) agonal status; (iii) time of death, and (iv) delay in tissue sampling; and GAD was found to be significantly influenced by (ii), (iii) and (iv) and CAT by (i), (iii) and (iv). None of these factors accounted for the total alterations in the enzyme activities of the mentally abnormal brains. The results indicate that reductions in cerebral GAD require to be interpreted with caution in view of the sensitivity of this enzyme to premortem status but that reductions in cerebral CAT may be a more reliable index of pathological change in senile (Alzheimer-type) dementia.


Immunocytochemical localization of the sodium, potassium activated ATPase in knifefish brain. Results presented in this paper demonstrate the feasibility of using immunocytochemical methods to localize the (Na+ + K+)-ATPase, and its subunits, in the C.N.S. We have shown that in the Black Ghost knifefish, Sternarchus albifrons, the enzyme is located on the plasma membrane of the somata and dendrites of neurons and on the somata and cellular processes of glia. In myelinated axons the enzyme is restricted in localization to those portions of the axolemma not covered by the myelin sheath. The capacity of cell plasma membranes to restrict mobility of functionally important proteins should be considered in models of membrane structure in which lateral mobility of membrane components is considered a major characteristic.


A 'late supernormal period' in the recovery of excitability following an action potential in muscle spindle and tendon organ receptors. 1. Discharge patterns have been recorded from five types of stretch receptor; frog muscle spindles, lizard tendon organs, cat soleus tendon organs and primary and secondary endings of cat soleus muscle spindles.2. The fully adapted discharge of each type of receptor is irregular, especially for frog spindles and primary endings of cat spindles as compared with the other three types (the ;regularly firing' receptors). Frog spindles and some cat spindle primary endings would maintain a discharge at very low mean rates (1/sec or less) while the remaining receptors would stop suddenly, as soon as their rate of discharge fell below a critical value characteristic for each individual ending.3. This pattern of discharge suggests that there is a peak in the excitability of ;regularly firing' receptors at a time following a preceding impulse, which corresponds to the intervals between impulses at each particular receptor's slowest rate of maintained firing, and that the excitability subsequently falls again. Primary endings of cat muscle spindles also showed some evidence of such a ;late supernormal period', but frog spindles did not.4. Direct evidence for the ;late supernormal period' was obtained from experiments in which a maintained discharge was restarted by an antidromic action potential in a receptor which had stopped firing, and to which had been applied a stretch just too small to restart the discharge.5. It is shown in an Appendix that a model receptor in which the recovery of excitability following an impulse has a hyperbolic time course, and in which Gaussian distributed noise is superimposed on the generator potential, can have a discharge pattern very closely resembling that of a frog spindle (cf. Buller, 1965).6. After addition of a late supernormal period to the model, its discharge pattern could mimic closely that of a lizard or cat tendon organ, or of a secondary ending of a cat spindle.


The close correlation between symptoms and disease expression in HLA B27 positive individuals. Following demonstration that 20 percent of presumed "healthy" HLA B27 positive individuals develop symptomatic ankylosing spondylitis, a controlled follow-up assessment of the remaining "asymptomatic" 80 percent was performed. The clinical and radiological study revealed that there is a close correlation between symptoms and radiologic change in HLA B27 positive subjects; those individuals remaining symptom free have normal pelvic radiographs. Ankylosing spondylitis or "asymptomatic sarcroiliitis" does not exist in a subclinical manner throughout the entire group of B27 positive subjects. Evaluation of the pelvic radiographs of both symptomatic and asymptomatic HLA B27 positive subjects and symptomatic HLA B27 negative controls demonstrated that osteitis pubis and fluffy periostitis are equally distributed among the three groups, only the frequency of sacroiliitis being statistically greater in the B27 positive symptomatic subjects.


The prune belly anomaly: heterogeneity and superficial X-linkage mimicry. The genetic, clinical, and necropsy findings of 2 brothers with the prune belly anomaly are presented and the literature reviewed. The combined data emphasise the clinical and genetic heterogeneity of the disorder and show that in at least some instances a heritable component may be the primary insult. The most likely heritable explanation involves a two-step autosomal dominant mutation with sex-limited expression that partially mimics X-linkage.


Regular G21-trisomy in 3 sibs from mother with trisomy 21 mosaicism. This paper describes a family with 3 affected sibs with regular trisomy 21 Down syndrome. The condition seems to be transmitted from a phenotypically normal mother in whom G-trisomy mosaicism was identified. Giemsa banding depicted trisomy 21 mosaicism in cells from the mother. Chromosomes from the children showed a trisomy 21 in all the cells analysed.


Serum cholinesterase (pseudocholinesterase) in Down's syndrome: 1. Phenotype frequencies at the E1 and E2 loci. Dibucaine, fluoride and RO2-0683 inhibition studies were used to determine the serum cholinesterase (pseudocholinesterase) phenotypes at the E1 locus in a sample of 130 subject with Down's syndrome and fifty-three mentally retarded control subjects. No example of the Ef1 and Es1 allele was detected in either group, nor were any of the genotype E1a E1a detected. The gene frequency of E1a for the control group (0.0189) resembles closely that reported in the literature for normal European populations but in the Down's group was significantly lower (0.0038). The means and distribution of the dibucaine, fluoride and RO numbers in both groups were similar to those reported in normal subjects. The presence of the additional C5 cholinesterase type heterozygous for a variant cholinesterase on the E2 locus) was detected after starch gel electrophoresis, and the frequency was found to be raised in both groups. Several possible environmental factors (age, sex, maternal age, etc.) were investigated to account for this finding but with no success.


Serum cholinesterase (pseudocholinesterase) in Down's syndrome: 2. Quantitative levels. The observation elsewhere (Drew and Rundle, 1977) that increase frequencies of the C5 + variant of the serum cholinesterase in Down's syndrome may be due to a protective influence against adverse environmental factors has been investigated for such factors as age, sex, duration of institutionalisation, presence of the hepatitis -B antigen and maternal age. With the exception of the maternal age none of the factors tested appear to affect the circulating levels of cholinesterase. A maternal age effect in the Down's subjects was detected with lower levels of the enzyme being found in the subject positive for the C5 + variant born to mothers over thirty-five years when compared to the C5 + subjects born to mothers under thirty-five years. Further studies confirmed the presence of a relationship between maternal age, serum cholinesterase levels and haptoglobin phenotypes.


The use of the Vineland Social Maturity Scale, the Merrill-Palmer Scale of mental tests (non-verbal items) and the Reynell Developmental Language Scales with children in contact with the services for severe mental retardation. Psychological tests were administered to a complete population of severely retarded children aged 0-14, from one area of south-east London. The fifty-six children selected for the present study included all those who had obtained scores on measures of social maturity, visuo-spatial skills not involving symbolic concepts, and level of language comprehension. The results show very low correlations between the age-related quotients obtained for each measure. This suggests that, in severely retarded children, marked discrepancies can occur between different areas of cognitive and social development. Some children could be classified as moderately or mildly retarded on one type of test but as profoundly retarded on another. The profiles on the tests can be related to diagnosis and behaviour pattern. The findings highlight the problems of assessment and educational placement of retarded children.


Thymus and breast cancer--plasma androgens, thymic pathology, and peripheral lymphocytes in myasthenia gravis. Plasma androgen sulfates were measured in 92 patients with myasthenia gravis. Plasma androgen sulfates were strongly associated with thymic pathology. The presence of germinal centers was associated with decreased androgen levels. A weak but significant positive correlation existed between androgen sulfates and lymphocyte counts. Low lymphocyte counts in patients with germinal centers were associated with subnormal androgen concentrations. Plasma androgen sulfate levels decreased immediately after thymectomy, but overall no significant differences were found between prethymectomy and post thymectomy levels. A trend toward higher concentrations of plasma androgens was noted 5 or more years post thymectomy. Plasma androgen sulfate levels in myasthenia gravis patients with breast cancer were markedly depressed. The lowest androgen sulfate levels were observed in patients who had previous bilateral breast cancer and those who, subsequent to the determination of the steroid sulfates, developed a second primary lesion of the breast.


Lack of involvement of monoamine oxidase inhibition in the lethality of acute poisoning by chlordimeform. Depletion of serotonin or norepinephrine in male rats by pretreatment with p-chlorophenylalanine or DL-alpha-methyl-p-tyrosine, respectively, failed to reduce the lethality of the monoamine oxidase-inhibiting acaricide, chlordimeform. Administration of phenylephrine, a directly acting alpha-adrenergic agonist, with  chlordimeform did not increase the lethality of chlordimeform. Thus, monoamine oxidase inhibition by chlordimeform does not seem to contribute to its lethality.


A carbon dioxide flowmeter.

A carbon dioxide flowmeter. A simple and inexpensive uroflowmeter has been designed that uses a common carbon dioxide cystometer. The patient voids urine in a specially designed container connected at the bottom of the carbon dioxide source in the cystometer. After proper calibration (500 cc fluid for a 20 mm. deflection in the paper) the patient is asked to void while carbon dioxide flows through the container and the paper recording commences. Any change of volume in the container produces a change in the pressure recorded by the cystometer, which will allow us to know, following calibration, the 2 factors involved in flow: 1) volume voided and 2) time. A table of average flow is available for a quick reading of the findings only by superimposing the table on the uroflow study. The only attachment needed for the cystometer is the special container. Our results support the use of this economical new way to determine urinary flow.


Metabolic changes with inflammation induced by a surfactant. Studies were carried out to determine the relationship between metabolic histopathological changes in the gastrocnemius muscle of rats in which an acute exudative inflammation had been induced by alkyldimethylbenzylammonium chloride (alkyl-DBAC, a cationic surfactant). Succinate respiration, Na+-K+-Mg2+ ATPase activity and ATP, ADP and AMP levels were determined as the index of metabolic changes. Myofascial edematous swelling with the acceleration of succinate oxidation and Na+-K+-Mg2+ ATPase activity was noted at 30 minutes in the inflamed muscle. The ATP level was also transiently reduced. On the other hand, Na+-K+-Mg2+ ATPase activity and succinate oxidation were inhibited by alkyl-DBAC, at this concentration, in vitro. These results support the possibility that enhancement of energy metabolism is not directly initiated by alkyl-DBAC but may be the result of certain chemical mediators released by alkyl-DBAC. The enhancement of energy metabolism continued after 1 hour and this energy may initiate leukocyte migration as well as increase vascular permeability and edema.


[Mathematical model of transient long-term adaptation processes of the heart]. A mathematical model of the heart's long-term adaptation to changing load is described. The model is based on the long-term adaptation concept which assumes the phosphorylation potential changing in the heart muscle cells with the load to control the genetic apparatus activity and thus determine structural alterations ensuring adaptation. The computerized model reproduces the transient process in four major types of the heart's long-term adaptation, i.e. in a compensatory hypertrophy development and hypertrophy regression, as well as in the training and detraining states formation. The simulation results are in agreement with the available data and predict the phenomena that are not as yet substantiated by experiments.


[Activity of key enzymes of glycolysis in the rat liver during the development of experimental myocardial necrosis]. Hexokinase, glucokinase, and phosphofructokinase activity in supernatant hepatic fluid obtained by centrifugation of the homogenate at 20,000 g for 20 minutes was studied during the development of experimental necrosis of the heart muscle. The activity of these enzymes was lowest in the 12th and 24th hour following arterial occlusion. Phosphofructokinase and glucokinase activity returned to the initial level on the 6th and 9th days, respectively; hexokinase activity was still diminished after the 12th day.


[Role of membrane-bound calcium in the changes of ion permeability and Na+, 5+ and ATPase activity of the erythrocytes in hypertension]. The content of calcium bound to the erythrocyte membrane and the effect of intracellular calcium concentration on the activity of Na+, K+-ATPase in the reconstituted erythrocytes were studied in 20 patients with essential hypertension and in 20 individuals with normal pressure. In incubation of the erythrocytes in a solution containing EDTA much more calcium is removed from the outer surface of their membrane in patients with essential hypertension than in the control group (60 +/- 5 mEq/l and 41 +/- 3 mEq/l, respectively). When the intracellular calcium concentration varies from 0 to 500 mumol/l, which corresponds to a rise in the free calcium (Ca2f+-3) concentration to 41 mumol/l, a difference in the changes of Na+, K+-ATPase activity of the reconstituted erythrocytes is noted. When intracellular calcium concentration is 50 mumol/l (Ca2f+-3 mumol/l), ATP-ase activity in patients with essential hypertension is 21% less than that in individuals with normal pressure (P less than 0.005). The authors explain the difference in the kinetics of Na+, K+-ATPase changes by the different degree of calcium depletion of the inner surface of the erythrocyte membrane in relatively low Ca2f+ values in the internal medium. The data obtained are evidence of the altered calcium-binding capacity of the erythrocyte membrane, which may cause the increased permeability of the erythrocyte membrane to sodium and potassium ions in patients with essential hypertension, which the authors had revealed earlier. The authors consider the revealed changes to be a fragment of a more extensive membrane defect which may be the principal cause of activation of the servomechanisms which maintain arterial pressure.


[Cardiac effects of antikaliuretic diuretics-clinical and biochemical investigation (author's transl)]. Recently direct myocardial effects of antikaliuretic diuretics with respect to contractility parameters and prevention of digitalis-induced arrhythmias were published. In order to test the value of these reports we measured the effect of potassium-canrenoate and triamterene on cardiac output and on digitalis-induced arrhythmias in patients during diagnostic and the therapeutic flow directed right heart catheterization (Swan-Ganz) in our intensive care unit. In addition the influence of these drugs on (Na+ + K+)-ATPase and on (3H)g-strophanthin binding to human cardiac cell membranes was investigated to gain information on the mechanism of their action. Triamterene (100-200 mg p.o.) was without any effect on cardiac output, the same was found true for potassium-canrenoate given in a single dose (200-1000 mg intravenously). However, when applied in two doses (200 mg i.v. and 60 min later 400 mg i.v.), potassium-canrenoate increased cardiac output by 11 percent (p less than 0.05). Only in 2 out of 14 patients potassium-canrenoate (200-400 mg i.v.) suppressed digitalis-induced ventricular ectopic beats. Canrenone, the active metabolite of potassium-canrenoate displaces [(3H)]g-strophanthin from its binding sites in human cardiac cell membranes and inhibits (Na+ + K+)-ATPase activity. These in vitro effects were measured at the same concentrations as found in vivo after "therapeutical" doses. The effects of triamterene in this respect were found only in extremely high concentrations. Our results imply that canrenone has cardiac glycoside-like effects in human cardiac cell membranes.


Murine lymphocytic choriomeningitis: the history of a natural cross-infection from wild to laboratory mice. A new locus of wild house mice tolerantly infected with the virus of lymphocytic choriomeningitis (LCM) has been identified in the United Kingdom. Evidence is presented which indicates that these mice were the source of infection in a laboratory mouse breeding colony, the mode of transmission probably being bites on tails and limbs exposed through wire-grid flooring. The results of an experiment which simulated indirect exposure of SPF mice to tolerantly infected wild mice supported earlier observations that without injury to the epidermis the risk of spread of infection from the infective urine, saliva or faeces is low.


The use of biotyping of Enterobacteriaceae as an evaluation of the isolation efficacy of laboratory animals. 2 sets of data are given of biotyping the Enterobacteriaceae from the faecal flora of SPF rats and cats housed with different isolation arrangements. The method is presented as an evaluation of the isolation efficiency of laboratory animals. Biotypes which were determined on almost every occasion belonged to the resident flora of the animals while those found occasionally represented contamination. Contaminating bacteria are cleared from an SPF breeding unit by colonization resistance and the continuous efflux of animals from the unit. A parameter of isolation efficacy is the number of different biotypes determined in time. With optimal isolation arrangements a stable bacterial flora of permanently colonized biotypes is obtained in the animals.


Alkaloids of Thalictrum. XXII. Isolation of alkaloids with hypotensive and antimicrobial activity from Thalictrum revolutum. Sixteen alkaloids were characterized from Thalictrum revolutum DC., namely; thalidasine, O-methylthalmethine, O-methylthalicberine, thalrugosaminine, thalicarpine, thalmelatine, pennsylvanine, palmatine, berberine, thalifendine, columbamine, jatrorrhizine, deoxythalidastine, thalphenine and magnoflorine. The structure of thairugosaminine (1) a bisbenzylisoquinoline type which was previously proposed on partial data was completely established, including the absolute configuration as S,S. Thalphenine, thalidasine, O-methylthalicberine, thalicarpine, thalrugosaminine and thaliglucinone were found to possess hypotensive activity in rabbits. Thalrugosaminine, thalicarpine, thalmelatine, O-methylthalmethine, pennsylvanine and thalphenine were found to be active against Mycobacterium smegmatis.


[Bacteriophage typing of Staphylococcus aureus strains isolated from patients in the microbiology department]. 78 Staphylococci isolated from mouth, throat, nose, ear passages and skin lesions were typed with 22 basic phage types. 68 of 78 staphylococci were typed. 45 of these (66%) could be typed with RTD, and 23 (33.8%) with 1000 X RTD. When phage grouping is considered, 14 out of 68 strains were from Group III and 12 out of 52 strains isolated from nose were from Group I phage.


[Bacteriophage typing of Staphylococcus aureus isolated from dental plaque and saliva]. Samples from 1127 persons were collected, S. aureus were isolated from 39 (3,4%) dental and from 4 (3,5%) saliva. The strains were tested with 22 basic phage types and 56 of them could be typed. 19 of them were typed by RTD and 19 by 1000 X RTD. Most of them were from phage Group II.


[Therapy of androgenization symptoms: double blind study of an antiandrogen preparation (SH B 209 AB) against neogynon (author's transl)]. The antiandrogen-containing preparation SH B 209 AB was tested in comparison with an ovulation inhibitor with no antiandrogenic effect (Neogynon) in a randomized, double blind study conducted to determine the effects of both on androgenization symptoms. 88 patients took part in the study. In the majority of cases the duration of the treatment period was more than 6 months. It was shown that the therapy with the combination preparation containing cyproterone acetate was distinctly more successful in cases of acne and seborrhoea: not only acne in its various locations but also all forms of seborrhoea were more favourably influenced by the trial preparation. This difference was statistically significant (p less than 0.05). It was not possible to establish a significant difference between the two preparations in the case of hirsutism in its various locations from the limited number of cases observed. There were also no differences detected in hepatic tolerance. During treatment with both preparations all values measured were within the normal range.


Nuclear-extranuclear interactions affecting oligomycin resistance in Aspergillus nidulans. The extranuclear mitochondrial oligomycin-resistant mutation of Aspergillus nidulans, (oliA1), was transferred asexually into four nuclear oligomycin-resistant strains of different phenotypes. In all four cases, the possession of the nuclear plus extranuclear mutation led to an increase in the in vivo level of oligomycin resistance. In two cases, the altered cytochrome spectrum and impaired growth ability determined by (oliA1) were suppressed by the nuclear mutations. In the third case, the in vitro oligomycin resistance of the double mutant ATPase was dramatically increased above that of either of the component single mutant strains, indicating a synergystic interaction between the nuclear and extranuclear gene products. In the fourth case, the double mutant became cold-sensitive. A new extranuclear mitochondrial oligomycin-resistant mutation (oliB332) is described. This mutant is phenotypically similar to, though not identical with, (oliA1) but is separable by recombination. A range of nuclear oligomycin-resistant mutants have been mapped. Despite presenting five distinctly different phenotypes, they all map at the same locus.


Interaction of the cytoplasmic membrane and ribosomes in Escherichia coli; altered ribosomal proteins in sucrose-dependent spectinomycin-resistant mutants. Alterations in the ribosomes of sucrose-dependent spectinomycin-resistant (Sucd-Spcr) mutants of Escherichia coli were studied. Subunit exchange experiments showed that 30S subunits were responsible for the resistance of ribosomes to spectinomycin in all Sucd-Spcr mutants tested. Proteins of 30S ribosomes were analyzed by carboxymethyl cellulose column chromatography based on their elution positions. Mutants YM22 and YM93 had an altered 30S ribosomal protein component, S5, and mutant YM50 had an altered protein, S4. Although a shift of elution position was not detected for all the 30S ribosomal proteins from mutant YM101, the amount of protein S3 was appreciably lowered in the isolated 30S subunits. A partial reconstitution experiment with protein S3 prepared from both the wild-type strain and YM101 revealed that the mutant had altered protein S3 which is responsible for the spectinomycin resistance. These alterations in 30S subunits are discussed in relation to the interaction between ribosomes and the cytoplasmic membrane.


Cholesterol in muscle membranes.

Cholesterol in muscle membranes. The composition of skeletal muscle microsomes is reviewed. Evidence for the involvement of cholesterol in the transport of calcium by vesicles derived from the sarcoplasmic reticulum is considered. Results obtained by non aqueous extractions of skeletal muscle microsomes, and by use of the cholesterol analogue 20, 25 diazacholesterol indicate that cholesterol is not involved in calcium transport by vesicles of sarcoplasmic reticulum origin. Use of density perturbation procedures indicating that cholesterol is present in muscle membranes other than those of the sarcoplasmic reticulum involved in calcium transport is discussed. The distribution of membranal cholesterol in muscle is compared to that in other tissues.


On the kinetics of acetylcholine at the synapse. Electrophysiologic, morphologic, and biochemical definitions of the synapse will be correlated spatially and temporally. Postsynaptic fatigue and facilitation follow oscillations of the free pool of acetylcholine, predicted by the kinetic theory and observed at the electric organ of Torpedo marmorata. The underlying thermodynamic instability exhibits properties of the Na+--K+-dependent hydrolysis of adenosine triphosphate and represents a necessary condition for synaptic memory.


Secretion of monoquaternary ammonium compounds by guinea pig small intestine in vivo. In anesthetized guinea pigs N-(3H)methylscopolamine (NMScop), N1-(14C)methylnicotinamide (NMN), and (14C)tetraethylammonium (TEA), administered intravenously, were secreted against a concentration gradient into the lumen of the small intestine. The concentration ratio of unmetabolized ammonium base in the intestinal lumen to that in the plasma was 4.3 and 6.5 for NMScop and NMN, respectively, 75 min after the intravenous injection of 1 nmole/g body weight of the individual compounds. The corresponding value for TEA after 180 min was 2.0. The establishment of the concentration gradient between intestinal lumen and plasma was diminished with increasing doses. An excess of NMN inhibited the uphill transport of NMScop. Since the electrical potential difference across the intestinal epithelium and a 'fluid circuit' mechanism cannot solely account for the observed accumulation of the monoquaternary ammonium compounds in the intestinal lumen, the evidence presented supports previous in vitro findings that the small intestine is capable of actively secreting organic cations.


Clinical neurogenetics. A survey of the relationship of medical genetics to clinical neurology. A retrospective survey was made of the prevalence of neurologic disorders with an established or possible genetic component in a university hospital patient population. More than half the patient visits to the Medical Genetics Clinic concerned diseases of the nervous system. More than 20 percent of pediatric neurology inpatients and at least 8.5 percent of adult neurology inpatients had disorders with a genetic factor. This study documents the common ground shared by medical genetics and clinical neurology.


An outline for the analysis of dementia. The memory disorder of Huntingtons disease. Methods have been developed for assessing the cognitive parameters contributing to a memory disorder. Our findings suggest that individuals with Huntington disease have impairments in the encoding of new information and the consistent retrieval from storage of learned material. Their difficulties lie particularly in the realm of episodic memory.


Unexplained infertility. A reappraisal.

Unexplained infertility. A reappraisal. During a 30-month period 229 couples were evaluated for infertility at the Naval Regional Medical Center, Oakland. Laparoscopy was used in all cases of otherwise unexplained infertility. The resulting laparoscopic examination of the female partner of 24 such couples demonstrated abnormal findings in 18 (75%). Of these 18 subjects, unsuspected endometriosis was found in 11 (46%) and peritubal adhesions in 7 (29%). Of the 229 couples evaluated, only 8 (3.5%) failed to show some etiologic factor associated with infertility. Thus, the previously reported 10 to 20% incidence of unexplained infertility is too high an estimate in view of the additional information made available through pelvic endoscopy.


Hormone levels following wedge resection in polycystic ovary syndrome. A study of serum estradiol, progesterone, 17alpha-hydroxy-progesterone, testosterone, dihydrotestosterone, dehydroepiandrosterone (DHA), delta4-androstenedione (delta4-A), FSH, and LH was carried out in one of three sisters having polycystic ovarian disease for a period of 18 days before wedge resection, at the time of surgery, and for 24 days following wedge resection. The mean levels of 17alpha-hydroxyprogesterone, testosterone, DHA, delta4-A, and LH were remarkably elevated prior to wedge resection. There was considerable day-to-day variation. Serum LH varied from 12.5 to 70.5 mIU/ml with a mean of 41.03 +/- 3.55 mIU/ml. Serum estradiol and progesterone levels were generally higher than those found in the early follicular phage. Wedge resection resulted in a fall in serum estradiol, progesterone, 17alpha-hydroxyprogesterone, DHA, and delta4-A. Ovarian secretion of the last four steroids was confirmed by a study of the ovarian vein blood obtained at the time of surgery. An estradiol peak occurred on the 14th post wedge resection day with smaller increases in 17alpha-hydroxyprogesterone, DHA and delta4-A. An increase in serum LH occurred on the 15th post wedge resection day with a peak on Day 16 accompanied by increases in FSH and progesterone. The postovulatory rise of progesterone was accompanied by reduction of serum LH levels to those generally found in the early part of the menstrual cycle. Various hypotheses for the ovulatory failure are discussed.


Differences between functional versus organic low back pain patients. There are contradictory findings about whether there are differences in personality and/or amount of emotional disturbance between patients with functional versus organic low back pain. The MMPI scores of 42 back pain patients diagnosed as "organic" were compared with the scores of 37 patients classified as "functional". The functional patients scored significantly higher than the organics on the Hs, Hy, Pd, Sc, Ma, and Si scales. The organics scored significantly higher than the functionals on the K scale. The results confirm the view that certain symptoms of emotional disturbance are more characteristic of patients who have relatively little evidence of physical findings. However, the degree of overlap between groups was high enough to suggest caution in making predictions and diagnoses about functional versus organic pain on individual patients solely on the basis of personality data.


Non-quantal release of transmitter at mouse neuromuscular junction and its dependence on the activity of Na+-K+ ATP-ase. The postjunctional membrane of mouse diaphragm fibres was hyperpolarized by 1-2 mV during local curarization of the end-plate zone in the presence of anticholinesterase. In a solution containing 5 mM K+, the mean hyperpolarization was 1.0+/-0.35 mV. Hyperpolarization was 1.46+/-0.30 mV when the activity of the Na-K pump was blocked by 2 x 10(-5) M ouabain and 1.82+/-0.43 mV when it was blocked by bathing the muscle in a K+-free medium. Reactivation of membrane ATP-ase by addition of potassium after a period in K+-free medium reduced the hyperpolarization to zero, when measurements were made 10-20 min after the readdition. It is concluded that spontaneous non-quantal leakage of acetylcholine occurs at the mouse neuromuscular junction, as it does in the frog (2). Conditions which block the Na+-k+-dependent ATPase increased the leakage whereas potentiation of the pump activity decreased it.


Studies on the biological role of DNA methylation: III Role in excision of one-genome long single-stranded phi X 174 DNA. Accumulation of replicative intermediates of the bacteriophage phi X174 was observed in E. coli C infected cells when phage DNA methylation has been inhibited by nicotinamide or when cells were infected with a temperature-sensitive mutant in gene A. Analysis of the accumulating replicative intermediates by electron microscopy revealed that these molecules are composed of double-stranded DNA rings with multiple-genome length single-stranded "tails". These results suggest that the single 5-methylcytosine residue present in the phage DNA serves as a recognition site for the gene A protein mediating the excision of one-genome long phage DNA. This excision process is oligatory for the final maturation of the phage.


[Role of iodine and manganese in the etiology of endemic goiter in the Martuksk region of the Aktubinsk district]. Iodine deficiency in the external environment served as the main etiological factor of endemic goiter. Low iodine content in the soil (1.8-390 mg/kg) undoubtedly served as the cause of endemic goiter development. Water from the sources of the Martuksk district contained iodine in low concentration. Soils of the Martuksk district are characterized by an acute deficiency of mobile manganese. Manganese concentration in the arable soil layer (16.9-75.4 mg/kg) was regarded as reduced, this being characteristic of the foci of goiter endemia. Water manganese content (from 66 to 186 microgram/1, 99.45 +/- 10.6 microgram/1, on the average) was below the maximal permissible concentrations indicated in the International standard for drinking water. Iodine, manganese content in the soil, food products and in the water of the Martuksk district of the Aktyubinsk region indicated a reverse relationship between iodine and manganese content and the extent of endemic goiter spread.


Action of nucleotide phosphotransferase of Escherichia coli on nicotinamide riboside and nicotinamide mononucleotide. The action of the nucleotide phosphotransferase of Escherichia coli on nicotinamide riboside and on its 5'-phosphate results in the addition of one phosphate moiety to each of the substrates. Although the proof is not conclusive, it is likely that the phosphate group is transferred to the 3'-hydroxyl of the ribose. This is in contrast to the behavior of the enzyme toward NAD in which only the adenylic acid portion is phosphorylated enzymically.


Migration of Escherichia coli dnaB protein on the template DNA strand as a mechanism in initiating DNA replication. The first step in conversion of varphiX174 singlestranded DNA to the duplex replicative form in vitro is the synthesis of a nucleoprotein intermediate [Weiner, J. H., McMacken, R. & Kornberg, A. (1976) Proc. Natl. Acad. Sci. USA 73, 752-756]. We now demonstrate that dnaB protein (approximately one molecule per DNA circle) is an essential component of the intermediate and retains its ATPase activity. Synthesis of RNA primers, dependent on dnaG protein (primase), occurred only on DNA that had been converted to the intermediate form. In a coupled RNA priming-DNA replication reaction the first primer synthesized was extended by DNA polymerase III holoenzyme into full-length complementary strand DNA. In RNA priming uncoupled from replication, multiple RNA primers were initiated on a varphiX174 circle. The single dnaB protein molecule present on each DNA circle participated in initiation of each of the RNA primers, which appear to be aligned at regular intervals along the template strand. We propose that dnaB protein, once bound to the template, migrates in a processive fashion along the DNA strand, perhaps utilizing energy released by hydrolysis of ATP for propulsion; in this scheme the actively moving dnaB protein acts as a "mobile promoter" signal for dnaG protein (primase) to produce many RNA primers. Schemes are proposed for participation of dnaB protein both in the initiation of replication at the origin of the Escherichia coli chromosome and in the initiation of primers for nascent (Okazaki) fragments at a replication fork.


Proof of de novo synthesis of the qa enzymes of Neurospora crassa during induction. In Neurospora crassa three inducible enzymes are necessary to catabolize quinic acid to protocatechuic acid. The three genes encoding these enzymes are tightly linked on chromosome VII near methionine-7 (me-7). This qa cluster includes a fourth gene, qa-1, which encodes a regulatory protein apparently exerting positive control over transcription of the other three qa genes. However, an alternative hypothesis is that the qa-1 protein simply activates preformed polypeptides derived from the three structural genes. The use of density labeling with D(2)O demonstrated conclusively that the qa enzymes are synthesized de novo only during induction on quinic acid. Native catabolic dehydroquinase (5-dehydroquinate dehydratase; 5-dehydroquinate hydro-lyase, EC 4.2.1.10) (a homopolymer of ca 22 identical subunits) has a density of 1.2790 g/cm(3) as determined by centrifugation in a modified cesium chloride density gradient. Growth in H(2)O followed by induction in 95% D(2)O shifts the density of the enzyme to 1.3130 g/cm(3), indicating de novo synthesis during induction. In the reciprocal experiment, i.e., growth in 80% D(2)O followed by induction in either 95% D(2)O or H(2)O, the densities of catabolic dehydroquinase were 1.3135 and 1.2800 g/cm(3), respectively. Because growth on D(2)O does not affect the density of the H(2)O-induced enzyme, there can be no significant synthesis of catabolic dehydroquinase prior to induction. Similar results were obtained for a second qa enzyme, quinate dehydrogenase (quinate:NAD(+) oxidoreductase, EC 1.1.1.24). Thus, induction of two qa enzymes involves de novo protein synthesis, not enzyme activation or assembly.


Oligomycin-dependent ionophoric protein subunit of mitochondrial adenosinetriphosphatase. A proteolipid isolated from yeast mitochondrial adenosinetriphosphatase (subunit 9) (ATP phosphohydrolase; EC 3.6.1.3) by chloroform/methanol extraction has been shown to discharge photo-induced potentials across a planar phospholipid membrane containing bacteriorhodopsin. Oligomycin, a specific inhibitor of oxidative phosphorylation which binds to this protein, allows the potential gradient to be reestablished. When proteolipid was isolated from an oligomycin-resistant strain, ionophoric activity was still obtained but the effect was not reversed by oligomycin. These studies suggest that the hydrophobic subunit-9 polypeptide is the ionophoric component linking ATP synthesis (hydrolysis) with proton translocation.


F1-ATPase-catalyzed synthesis of ATP from oleoylphosphate and ADP. Purified preparations of F1-ATPase (ATP phosphohydrolase; EC 3.6.1.3) isolated from yeast mitochondria catalyze the reaction of oleoylphosphate with ADP to yield ATP and oleic acid. Formation of ATP is specifically inhibited by the F1-ATPase inhibitor 1799 and by dinitrophenol. In the presence of F1, dinitrophenol "uncouples" the synthase reaction by causing rapid hydrolysis of oleoylphosphate without ATP formation. It is proposed that this F1 catalyzed ATP synthesis reaction corresponds to the terminal chemical step in oxidative phosphorylation.


Identification of dynein as the outer arms of sea urchin sperm axonemes. The location of dynein, the main flagellar ATPase, within the sea urchin sperm axoneme was investigated by the use of immunofluorescence and immunoelectron microscopy, employing an antiserum against a tryptic fragment of dynein 1 (Fragment 1A) purified from sea urchin sperm flagella. The axonemes were found to be stained with the antiserum when examined by an indirect immunofluorescence technique. Immunoelectron microscopy with the antiserum and a ferritin-conjugated IgG fraction of goat antiserum to rabbit IgG revealed that, among the structures within the axoneme, only the outer arms were labeled with ferritin particles. With either the normal serum or antiserum absorbed with Fragment 1A, there were no ferritin particles within the axonemes. When the outer arms were extracted with 0.5 M NaCl, leaving the inner arms intact, again no ferritin dots were detected. Furthermore, it was found that the outer arm on the no. 5 doublet microtubule, which connects with the extra arm projection backward from the no. 6 doublet, had no attached ferritin particles. From these observations, it can be concluded that the outer arm consists of dynein (at least dynein 1) and that Fragment 1A, containing the active site for ATPase activity of dynein 1, is located at the distal end of the outer arms. The significance of the present findings is considered in connection with flagellar movement.


Histological investigation of aortic wall in experimental sclerosis of rabbits. Rabbit aortic changes were investigated after different sclerogenic diets. A subintimal chondroitin sulfate layer was characterized by toluidine blue metachromatic staining at pH 1--3, CEC value expressed in MgCl2 concentration: 0.4M, hyaluronidase sensitivity. This layer becomes disorganized during plaque formation and partly disaappears. A long-lasting sclerogenic diet, as well as a diet containing several sclerogenic factors, produced an increase of frequency and extension of the media necrosis, calcification, and chondroid metaplasia of the aortic wall. In these lesions, an increase of mucopolysaccharide secretion of modified smooth muscle cells was observed.


Antioestrogenic action of the aldosterone antagonist canrenoate K in the rat (adenohypophysis, ceruloplasmin). In a dose of 7,k mg/rat/day in food, the aldosterone antagonist canrenoate K inhibited the adenohypophyseal reaction (growth, raised thyroxine-binding capacity of the adenohypophyseal proteins in vitro) and the ceruloplasmin reaction (elevation of the serum ceruloplasmin level) to three weeks' intramuscular administration of long-acting oestradiol benzoate in doses of 1 mg twice a week. The effect was similar to that of the antioestrogen clomiphen in a dose of 1.25 mg/rat/day in food. In combined administration of clomiphen and canrenoate K, no summation of their effect was observed. Neither canrenoate nor clomiphen affected the post-oestradiol drop in body weight, but they both potentiated the oestradiol-induced decrease in testicular weight and canrenoate potentiated the effect of oestradiol on uterine weight. It was therefore concluded that the effect of canrenoate is not of a catatoxic nature, i.e. that it is not determined by increased metabolic degradation of oestradiol.


Effect of intravenously infused noradrenaline on the cardiovascular system of cold-acclimated rats. Total metabolism, body temperature and basic circulatory parameters (the heart rate, blood pressure, blood oxygen content and hind limb blood flow) were measured in anaesthetized cold-acclimated male laboratory rats exposed to the calorigenic action of noradrenaline. In addition, cardiac output, the oxygen extraction coefficient and peripheral vascular resistance were calculated from these data.


Pulmonary haemodynamics during nasal apnoea in rabbits. Changes in pulmonary haemodynamics, produced during stimulation of the nasal mucosa with xylol fumes, were found in experiments on anaesthetized rabbits. In spontaneously breathing animals, a decrease in the mean blood pressure in the pulmonary artery and an increase in the mean blood pressure in the left atrium occurred in addition to apnoea and bradycardia. Pulmonary vascular resistance fell significantly, whereas systemic vascular resistance rose. Changes in the pulmonary circulation during nasal stimulation in spontaneously breathing rabbits did not differ significantly from changes in artifically ventilated animals.


Effect of anaesthesia on the pattern of breathing. The pattern of breathing of male rats was studied after stimulating respiration with carbon dioxide at different levels of general anaesthesia. Anaesthesia was induced by the inhalation of halothane or by the i.p. injection of urethane. Ventilation values were measured in intubated rats in body plethysmograph. It was found that a linear relationship between minute ventilation and tidal volume was maintained during the decrease of minute ventilation due to deepening of anaesthesia. The slope of the relationship after stimulating respiration with carbon dioxide also diminished during deeper anaesthesia. The duration of inspiration did not alter significantly, despite marked changes in tidal volume. Tidal volume correlated with the duration of expiration at different anaesthesia levels. In vagotomized rats, the duration of expiration shortened as ventilation was depressed by deepening anaesthesia.


Development of spontaneous motility in chick embryos. Sensitivity to aminergic transmitters. The consequences of systemic administration of aminergic transmitters (n-adrenaline 16 microgram/kg egg weight; serotonin 2.5 and 5 mg/kg e.w.; dopamine 2.5 and 5 mg/kg e.w.) for the spontaneous motility and heart rate of 11- to 19-day chick embryos were studied intack eggs. The following results were characteristic for all three transmitters: a) when administered to 11- and 13-day embryos their effect was non-significant; the first signs of activity did not appear until the 15th day of incubation. The effect on 17- and 19-day embryos was stronger. b) After the 15th day of incubation, all these transmitters had a predominantly inhibitory effect on spontaneous motility; in 17- and 19-day embryos this acquired a periodic character. c) The changes in spontaneous motility did not correlate significantly in any way with the relatively small heart rate changes.  It is concluded from the results that aminergic mechanisms begin to participate in regulation of the spontaneous motility of chick embryos from the 15th day of incubation, and not before.


[In vitro studies to elucidate the fibrinolytic activity in cerebrospinal fluid altered by inflammation and hemorrhage and having pathologic protein values]. Using selectively immunological methods, it was possible, through FSP determination, for plasmin activities and plasminogen concentrations to be occasionally and exclusively detected in inflammatorily altered liquores and in bloody liquores, respectively. Thus, bloody cerebrospinal fluid, in contrast with inflammatorily altered liquor, usually shows free fibrinolytic activity, so that antifibrinolytic therapy of intracranial aneurysmal hemorrhage is pathophysiologically justifiable.


Evidence for a noradrenergic mechanism in the grooming produced by (+)-amphetamine and 4, alpha-dimethyl-m-tyramine (H 77/77) in rats. Rats were kept on a 12-h light-dark cycle. One hour after the light was switched on, physiological saline, (+)-amphetamine 1 mg/kg, and H 77/77 5 mg/kg were injected s.c.; the number of groomings was counted 1-2 h after the treatments. (+)-Amphetamine and H 77/77 produced increased grooming which was antagonized by the tyrosine hydroxylase inhibitor H 44/68 (250 mg/kg), the dopamine-beta-hydroxylase inhibitor FLA 63 (40), the neuroleptics haloperidol (0.1 and 0.5), and clozapine (1 and 5). The (+)-amphetamine-induced grooming was also antagonized by the NA-receptor blocker aceperone (10) but not by the sedative phenothiazines mepazine (10) and diphenhydramine (20) nor diazepam (1). These results indicate that NA-release is involved in the mediation of (+)-amphetamine- and H 77/77-induced grooming. The inhibition of haloperidol and clozapine is presumably due to NA-receptor blockade.


Circadian rhythms in lung resistance and dynamic lung compliance of healthy children. Effects of two bronchodilators. In two groups of healthy children synchronized with a diurnal activity (light-on at 07.00) and a nocturnal rest(light-off at 21.00), lug resistance (R1) and dynamic lung compliance (C1 dyn) were measured at fixed clock hours (07.30, 11.30, 16.30, 22.30). The measurements were performed before and 10 minutes after the inhalation of bronchodilators (a beta-sympathetic stimulating agent (orciprenaline, dose 2 mg) and a vagolytic agent (SCH 1000, doses 80 mug and 200 mug). A circadian rhythm is detected for R1 and C1 dyn and quantified by the cosinor method. R1 and C1 dyn acrophase (peak time in the 24-h scale) are not significantly different for both groups: they are located for R1 at 05.39 and 03.31, for C1 dyn at 09.31 and 12.51. R1 and C1 dyn circadian rhythms are not detectable both after 2 mg orciprenaline and 200 mug SCH 1000 inhalation. This chronopharmacologic effect of the bronchodilators might be considered indirect evidence of a circadian rhythm in the bronchial tone. This latter may be related to circadian changes in neurovegetative activity. A dose-effect difference of the vagolytic agent on R1 at night seems to indicate a relative nocturnal prominence of the vagal tone.


[Relations between human and animal influenza viruses]. The following more important problems are dealth with in the present paper:--antigenic relationships between human and animal influenza viruses;--recombination of human and animal influenza viruses;--epidemiological relationships between human and animal influenza viruses;--the theory of cyclical evolution of influenza viruses.


[Aspects of bacteriophage-eukaryotic cell relationships]. The present work deals with bacteriophage--eukaryotic cell interrelationship, including:--Data on the possible presence of bacteriophages in calf serum and viral vaccines.--Methodological problems of research on phage--eukaryotic cell relations.--Data concerning the effects of phage infection on eukaryotic cells (demonstration of phage infection of the eukaryotic cell; integration of phage genetic material in the genome of the eukaryotic cell; coding of phage proteins in phage-infected eukaryotic cells; modified multiplication parameters and transformed potential of some phage-infected eukaryotic cells).--Indications for solving some of the problems.


[Mucoproteinuria in complicated scarlet fever]. Mucoproteinuria, determined by Bugard's semiquantitative method, revealed values of 100-350 mg/24 h in common scarlet fever and of 300-882 mg/24 h in scarlet fever complicated by early nephritis, pseudorheumatism and acute diffuse glomerulonephritis. These alterations appear to lend support to the recent hypothesis concerning the mucoprotein substrate of the autoimmune mechanism in the complications of streptoccal infection.


[Aspects of the relation between Enterobius vermicularis and appendectomy in Oradea]. The relationship between the nematode Enterobius vermicularis and appendicectomy in three age-groups: 0--15, 16--20 and 21--25 years was followed during the 1968-1973 period. Following the compaign against enterobiosis, there was a significant decrease in the number of operations for appendicitis in the 0--15 years age-group; the incidence of appendicectomy was greater in the 16--20 years age-group than in the reference 21--25 years age-group.


[Simplified diagnostic scheme for Enterobacteriaceae]. After testing 18 experimental variants against 40 reference strains belonging to different Enterovacteriaceae groups, the authors chose a multitest medium that permits concomitant testing of mobility, indol, lysinedecarboxylase and phenylalanine deaminase (MILP). In this medium, the reactions of 3734 strains belonging to the genera Escherichia, Shigella. Salmonella, Citrobacter, Klebsiella, Enterobacter, Serratia. Proteus and Providencia were concordant with the reactions in individual test media, taking as reference, in a proportion of 99.7 and 100%. The authors, associating the MILP medium to TSI agar, propose a simple scheme consisting of eight test which permits, within only 24 hours, the correct identification of the genus and sometimes of the species of cultures or isolated colonies from selective media.


[Intrauterine inoculation, an experimental model in colibacillary infection in mice]. Impuber females of the RAP, Swiss, Albino-Rosso, NRMI and C57Bl mice strains were inoculated by intrauterine route with E. coli O55B5, following up the mortality rate, dissemination and persistence of the bacilli in different organs, the onest of possible lesions in the urinary and genital tract, the reproduction capacity and frequency of coli diarrhea in the progeny. The C57Bl, NRMI and H strains were more sensitive, and the RAP, Swiss and Albino-Rosso more resistent to the lethal effect of the intraperitoneal inoculations. Intrauterine inoculation revealed no significant difference in the susceptibility of the animal strains, the mortality rate being virtually nil even when doses much higher than those inoculated by i.p. route were used, except for C57Bl mice in which lethal cases were recorded also after intrauterine inoculation. E. coli was isolated from the liver and sporadically from the kidneys and urinary bladder up to 14 ways after i.p. inoculation. After intrauterine inoculation E. coli was isolated from the uterus up to 30 days, and sporadically from the kidneys and urinary bladder, but not from the liver. In this case, too, the C57Bl mice were the exception, E. coli benig found in the liver after both i.p. and intrauterine inoculation. The latter inoculation did not influence the reproduction capacity of the females or the appearance of epizootic diarrhea in the progeny.


Peritoneoscopy as a diagnostic supplement to liver function tests and liver scan in patients with carcinoma. Liver function tests, liver scintigraphy and peritoneoscopy were carried out in 240 patients with carcinoma. Hepatic invasion was demonstrated by peritoneoscopy essentially when both of the other tests were positive. However, in one-third of the patients, peritoneoscopy revealed the presence of other pathologic changes which could account for the positivity of either of the other diagnostic procedures. The status of the liver, with regard to presence or absence of metastases, could be microscopically documented in 59 patients. False-negative findings of liver chemistry tests, liver scan and peritoneoscopy were seen in 27, 42 and 36 per cent, respectively, of the patients while the rate of false-positive results was 15, 10 and 3 per cent, respectively. The rate of false-negative peritoneoscopy examinations in the absence of simultaneous positivey of the two other investigations was insignificant. These data indicate that liver chemistry tests, liver scan and peritoneoscopy play a major and complementary role in the screening of patients with carcinoma.


Distribution of water between extracellular and intracellular compartments of incised wounds of rabbits. The distribution of water between the extracellular and intracellular compartments in incised wounds of skin, muscle and stomach has been studied in healthy rabbits and the progress of healing monitored by the determination of tensile strength for 120 days. It has been shown that, after wounding, there is an immediate expansion of the extracellular space. The increase is most marked during the first 24 hours and is maximal by this time in wounds of the skin and the muscle. During the first 30 days of healing, all incised tissues contain similar amounts of extracellular water which constitute approximately one-half of the total tissue mass, irrespective of the size of the extracellular space prior to wounding. There is an inverse relationship between the increase in the amount of extracellular water and the size of the extracellular space prior to wounding. All incised tissues maintain an elevated, and similar, concentration of extracellular water for more than 120 days. The period of maximal gain in tensile strength corresponds to the period of maximal expansion of the extracellular space which signifies a particularly active phase in the wound healing process. The intracellular space in wounds of muscle and stomach is reduced for more than 120 days, but no significant changes are seen in the intracellular water of wounds of skin. It has been concluded that expansion of the extracellular space is essential for wound healing, and it can, therefore, serve as a sensitive indicator of tissue injury.


Celiac axis, superior mesenteric artery, and inferior mesenteric artery occlusion: surgical considerations. Occlusion of the celiac, superior mesenteric, and inferior mesenteric artery has been studied in 46 patients treated by operation. The condition was acute and was caused by embolic obstruction of the superior mesenteric artery in four cardiac patients and detachment of the inferior mesenteric artery in two patients during removal of infrarenal abdominal aortic aneurysms. The condition was chronic and involved two or all three of the vessels in 40 patient. Embolic obstruction caused severe abdominal pain but few physical signs early in the process,, but the picture of an acute abdomen indicating bowel gangrene developed in a few hours. Ischemia from inferior mesenteric detachment was observed at operation. Patients with chronic obstruction had abdominal pain, weight loss, and diarrhea. Patients with embolic obstruction were treated successfully by embolectomy, and patients developing intraoperative sigmoid ischemia were treated by reattachment of inferior mesenteric arteries to aortic graft. Various procedures were employed in patients with chronic multiple obstruction. However, graft bypass using Dacron tubing was preferable because of its simplicity and because the frequently (48%) associated occlusive disease and aneurysm of the distal aorta were treated at the same time. Confining operation to the abdomen significantly reduced the magnitude of operation and eliminated risks in this age group. Of the 46 patients, 91% survived and were relieved of their symptoms despite associated disease. The 5-year survival rate in this group of patients was 62%.


The reoperation of choice for aortofemoral graft occlusion. Restoration of circulation following occlusion of an aortofemoral graft was attempted in 50 limbs in 38 patients. Progressive arteriosclerosis in the femoral vessels was the cause of graft limb thrombosis in most instances. Profundaplasty was employed in 47 limbs and femoral-popliteal bypass in two to provide effective runoff. Inflow was established by thrombectomy in 30 of the 37 limbs in which it was attempted, and it was readily accomplished with a Fogarty catheter if carried out within a few weeks after graft occlusion. In the 20 limbs in which thrombectomy was either abandoned (seven or not feasible because of prolonged delay (13), new grafts were inserted (13) or extra-anatomic bypass was carried out by femoral-femoral (four) or axillary-femoral (three) techniques. Operative death occurred in one patient (2.6%) after transabdominal graft replacement, and only two other patients had significant complications with full recovery. Initial graft patency was achieved in 96.6% following thrombectomy and in 85% following new graft or extra-anatomic bypass. Cumulative patency was 75% after 3 years. Amputation was required in five of the limbs in which graft patency could not be maintained. Reoperation should be performed promptly after graft limb thrombosis. The procedure of choice is thrombectomy and profundaplasty through a simple groin incision.


Protective effect of drugs on histamine-induced asthma. Controlled standardised histamine inhalation tests were carried out in 21 asthmatics to determine the degree of non-specific bronchial hyperreactivity with and without prior treatment with several anti-asthmatic drugs. A significant protective effect was produced by inhaled salbutamol, 200 microgram, ingested salbutamol, 4 mg, inhaled Sch1000, 40 microgram inhaled atropine sulphate, 290 microgram, and ingested choline theophylinate (200 or 400 mg) producing serum theophylline levels over 10 mg/l. Inhaled salbutamol was consistently the most effective and was significantly better than the other drugs. The protective effect between the other four was not significantly different. Drug side-effects occurred only with the ingested drugs. No significant protection was detected after ingested choline theophyllinate producing serum theophylline levels of less than 10 mg/l, inhaled sodium cromoglycate, 20 mg given once or six-hourly for one week, or ingested ascorbic acid, 1 gram.


[Nuclear proteins and DNA in the myocardial myocytes in compensatory cardiac hypertrophy]. Using cytophotometry, the amount of DNA, total nuclear proteins and of histones were studied in the myocardial cells during days 21--36 of experimental compensatory hypertrophy of the heart (in rats). The enlargement of myocardial nuclei during the cardial hyperfunction was accompanied by accumulation of total nuclear protein, in particular, the histone fraction, without distinct changes in DNA. Analysis of correlations between nuclear proteins and DNA in the myocardial cells allows to reveal a delayed accumulation of histones in the big and gigantic nuclei, with a superfluous increase in non-histone nuclear proteins. In middle-sized nuclei, non-histone proteins have little changes against intensive accumulation of histones.


[Immunogenetic characteristics of the population of the Transcarpathian region of the UkrSSR ABO system erythrocyte antigen findings]. The phenotypes with their respective alleles frequencies of the ABO system were studied in 33 230 individuals of 9 ethnic groups of the Transcarpathian Region population. Statistically significant differences in allele frequencies were found in Gypsies, Germans and Slovaks as compared to those in the main Ukrainian population. There are significant differences between Hungarians and Gypsies of the Transcarpathian Region and analogous populations beyond the region. Absence of a reliable difference between gene pools of the Slav groups of the population and of Hungarians may point to the local origin of the later.


[Significance of the autoimmune component in the genesis of inflammatory "tumors" of the abdomen]. Data concerning  the studies on autoimmune processes in 110 cases of inflammatory "tumors" of the abdomen and its anterior wall are set forth in this article. With this goal, pathomorphological immunomorphological studies of inflammatory "tumors" specimens, a dynamic study of the reaction intensity of the complement fixation (RFC) with the tissue antigen and also the immunoglobuline  serum level together with eosinophylic reaction of the organism were carried out. The dynamic study of the fixation of the complement reaction, immunoglobuline level and eosinophylic reaction of the body permits to make a more complete judgement on the degree of the pathological process activity and on  the postoperative clinical course.


Hyperplastic and metaplastic lesions in the reproductive tract of male rats induced by neonatal treatment with diethylstilbestrol. Newborn male Wistar rats were castrated on the day of birth (= day 1) and treated daily with diethylstilbestrol (DES) from days 1 to 30; 1 microgram for the first 10 days of life, 2 microgram for the next 10 days and 4 microgram for the last 10 days. The animals were autopsied at 30, 90 and 270 days of age. The epithelium of the coagulating glands (CGs) and ejaculatory ducts (EDs) underwent metaplastic transformation in all DES-treated rats. These pathological changes were more marked with age. The most striking changes were found in the periurethral regions of the CGs and EDs and associated regions of the dorsal urethral wall. The normal transitional epithelial lining almost disappeared and large papillary epithelial outgrowths occurred near the opening of the EDs and CGs. This type of neoplastic change was most marked in the group of rats sacrificed at 9 months of age.


The influence of chronically hypoxemic states on human carotid body structure and cardiac hypertrophy. Quantitative and qualitative changes in the human carotid body morphology, and their relationship to changes in the weight of right and left ventricles were investigated in 10 patients with a history of chronic hypoxemia. 5 patients without a history of cardiac, pulmonary or cerebral respiratory failure served as the control group. In the chronically hypoxemic group, a 2.67-fold increase in the total specific glomus cell volume was found. Up to a critical volume this increase is due to hypertrophy, beyond that it is due to hyperplasia. The course of the morphologic changes under the influence of slowly progressive chronic hypoxemia is discussed in a frame work of three stages (stage I - hypertrophy, stage II = nodular hyperplasia, stage III = atrophy). Plasmacellular infiltrates are constant though sometimes sparse. They are mostly perineural in location, less often intralobular and if so almost exclusively periglomoidal. In one case, we found an increase of Schwann cells in the interstitial and periglomoidal space without demonstrable degeneration of the nerve fibres themselves. Our hypothesis suggests that degeneration of special nerve terminals of the reciprocal type occurs in afferent nerve fibers. The increase of right ventricular weight (by a factor of 2.05) is significant, in contrast to that of the left. A linear correlation between the increase of right ventricular weight and the increased total glomus cell volume was not established. In 4 cases, however, we found pulmonary hypertensive vascular changes, which might be responsible for the disparity in the linear relationship.


Ultrastructural study of a hitherto unknown disturbance of the synthesis of collagen precursor molecules in human cutis. The ultrastructural characteristics of on hitherto unknown disturbance of collagen precursor molecules in human skin were studied. A severe disturbance of the synthesis of collagen exists at the procollagen stage. The paracrystalline accumulation of a form of procollagen in the tubes of the smooth ER and an increase in its appearance in the extracellular space, distinguish this clinical picture from all other previously described connective tissue disorders, especially from Dermatosparaxis and Ehlers-Danlos syndrome type VII. For this clinical picture we suggest the name "Dermatosparaxis endoplasmatica".