Hospital follows two-phase plan for move into new building. For the move into its new facility, the management of this large general hospital assigned responsibility for master planning and hospital-wide functions to a central team and assigned responsibility for detailed tasks to individual departments and staff members. How members of the hospital's staff met special challenges, such as training staff members for new systems and maintaining continuous operation of its regional emergency-trauma center during the move, also is examined.


Hospital meets patient demand for 'home-style' childbirth. A university-affiliated hospital faced increasing patient demand to minimize or eliminate traditional medical and institutional involvement in labor and delivery. To develop appropriate responses to this demand, the hospital established a committee that included professional practitioners and consumers. Responses developed by the committee included a "home-style" labor and delivery room, more liberal visitation policies, and revised medical procedures. Through a series of compromises, the committee was able to meet patient demands while maintaining the benefits of medical and institutional care.


Work load management system ensures stable nurse-patient ratio. The concept of adjusting work load to staffing is the reverse of most currently accepted practices. However, that is precisely how this management and measurement system works. Patient care units (PCU'S) are estimated for all incoming admissions, and patients are assigned to the units that have the lowest PCU count, the goal being an even distribution of work load among nursing units.


Emergency/outpatient satellites serve as rural outposts. Physician recruitment problems and sudden population growth placed a severe strain on several New Hampshire communities until civic and hospital leaders pooled resources to develop combined emergency/outpatient satellites. Range of services, staffing, operations, financing, and impact of the satellites are discussed in detail.


Hospital, nursing homes become partners in psychiatric care. Hospital psychiatric staffs can effectively share their expertise with nursing homes. One hospital's psychiatric out-reach program has reduced patient management problems at participating nursing homes, improved patients and staff morale, changed staff attitudes, garnered physician support, and extended services provided by the psychiatric department.


Double translocation t(7;12),t(2;6) heterozygosity in one family. A contribution to the trisomy 12p syndrome. Double translocation heterozygosity t(2;6),t(7;12) in three generations of a Dutch family is described: the segregation of a double translocation in more than one generation has not been previously published. The index case was a 16-year-old mentally retarded boy with partial trisomy 12p who showed several dysmorphic features such as high prominent forehead, flat face, flat and short nose bridge, short nose, dysplastic ears, prominent lower lip, and several skeletal abnormalities. Based on the findings in this patient and those in nine other cases, the existence of a specific trisomy 12p syndrome is postulated.


Quantitative studies on the arrangement of human metaphase chromosomes. V. The association pattern of acrocentric chromosomes in human meningiomas after the loss of G and D chromosomes. The association pattern was studied in 2715 mitoses of 90 meningiomas with different numbers of acrocentric chromosomes. In cells with monosomy 22, a significant increase of mitoses with associations was observed in comparison to cells with a normal karyotype. The number of associating acrocentric chromosomes was highly significantly increased. This surplus was not only caused by a highly significant increase of associating G chromosomes but also of D chromosomes. The loss of further acrocentric chromosomes had no significant influence on the number of mitoses with associations or the number of associating chromosomes. Based on the well-known correlations between the nucleolus organization and the association pattern, the results seem to indicate a compensation mechanism among the nucleoles organizing regions (NOR's) which keeps the supply of nucleolar material constant and simultaneously causes a higher association tendency between the remaining acrocentric chromosomes. The increase of associations in the 22 monosomic cells was interpreted as a overcompensation after the loss of only one NOR.


Heterogeneity of glucose-6-phosphate dehydrogenase deficiency in Algeria. Study in Northern Algeria with description of five new variants. Glucose-6-phosphate dehydrogenase (G6PD) deficiency was found in 3.2% of the male population living in the urban area of Algiers. The deficient subjects originated from multiple geographic regions of Northern Algeria, with prevalence of individuals of Berber-Kabyle origin. Red blood cell G6PD was partially purified and characterized in deficient males from 17 families, and six different variants were found. Among them, only one, the Gd(-) Kabyle variant, had been previously described. It was detected in nine families. The other five variants were new: Gd(-) Laghouat (four cases), Gd(-) Blida (one case), Gd(-) Thenia (one case), Gd(-) Titteri (one case), and Gd(-) Alger (two brothers). Strikingly, the common Mediterranean variant was not found. G6PD deficiency is heterogeneous in northern Algeria where autochtonous variants seem to prevail. The Kabyle variant may be common in this country.


Effects of caffeine on sister chromatid exchange (SCE) after exposure to UV light or triaziquone studied with a fluorescence plus giemsa (FPG) technique. Studies are reported that are designed to analyze the mechanism by which caffeine reduces the induction of SCE by UV light or alkylating agents. The substantial points are (1) caffeine does not inhibit SCE formation, and (2) the caffeine-mediated apparent decrease of SCE induction after UV or triaziquone exposure is due to a selective destruction of those metaphases otherwise exhibiting a high number of SCE. These findings and their relevance to the ascertainment of the SCE-forming process are discussed.


Monogene inheritance of learning speed in DBA and C3H mice. A behavioral genetic study in the shuttle-box. We carried out investigations on C3H, NMRI, C57Bl/6, Balb/c, Balb/cN, and DBA inbred mouse strains in the shuttle-box to see whether their learning behavior is genetically controlled. The highly different learning behavior of the parental strains made it possible to test the F1 hybrids and the F2 generation. The environmental influences were standardized as much as possible. In particular, influences possible during the lactation period were excluded by using foster breeding. The results enable us to postulate monogenic inheritance for the learning speed in the shuttle-box. The inheritance is interpreted as codominant. The investigations are part of a basic study in mammalian behavior genetics from the human genetic aspect.


Human liver alcohol dehydrogenase isoenzyme variations. Improved separation methods using prolonged high voltage starch-gel electrophoresis and isoelectric focusing. Human liver alcohol dehydrogenase isozyme patterns were studied using prolonged high voltage starch-gel electrophoresis and gel-slab isoelectric focusing. Homo- and heterodimers of ADH2 locus were easily distinguished from each other. The gene frequencies of ADH2 2 and ADH2 3 in 46 random liver samples from Germany were found to be 0.044 and 0.424 respectively.


Successive spontaneous abortions including one with whole-arm translocation between chromosomes 2. A family with five induced and seven spontaneous abortions and no live births is described. Four of the seven spontaneous abortuses were available for cytogenetic examination and three were successfully karyotyped. Their karyotypes were 46,XX; 46,XX/46,XX,t(2;2)(2p2p;2q2q); and 46,XY. The karyotypes of the parents were normal. The origin of the 2p/2p and 2q/2q translocation in one of the abortuses was assigned to an interhomologous whole-arm translocation in an early mitotic division in a conceptus with a 46,XX karyotype.


Isochromosome for the long arm of the Y in an infertile male. A 37-year-old man investigated for infertility had bilateral atrophic testes. Cytogenetic investigations revealed a chromosome complement of 45,XO/46,Xi(Yq)/46,XY. Mechanisms for the origin of the i(Yq) are considered, and the relation of his chromosome constitution to his infertility and hypogonadism are discussed.


Autosomal recessive sensorineural-conductive deafness, mental retardation, and pinna anomalies. Three siblings, a boy and two girls aged 11, 9, and 4 years, were found to have a congenital disorder characterized by malformed, low-set ears and sensorineural-conductive hearing loss. Variable expressivity was evident, since the boy had both types I and III of microtia and his two sisters had only type I. The normal parents were third cousins. The analysis of these findings permits the identification of a distinct nosologic entity due to the homozygocity of an autosomal recessive mutation.


Development of improved media and culture conditions for clonal growth of normal diploid cells. Multiplication of normal diploid cells in culture is controlled by a complex set of interacting extracellular variables. The amount of serum protein needed for colony formation by such cells is affected directly by many of the other variables, including the nature of the culture surface, the type of trypsinization procedure used, and the qualitative and quantitative composition of the culture medium. By a sequential process of adjusting all of these variables to optimum values for cellular multiplication with minimal amounts of serum protein, we have been able to obtain clonal growth of normal human and chicken cells with less than 500 microgram per ml dialyzed serum protein. Precise quantitative adjustment of nutrient concentrations is particularly important. The multiplication-promoting functions of serum can be classified operationally as "replaceable" (those that can be replaced by modifying the medium or the culture conditions) and "nonreplaceable" (those that we have not yet been able to replace). Elimination of the requirement for replaceable functions of serum has improved greatly the specificity and sensitivity of the bioassay for the nonreplaceable functions. The nonreplaceable multiplication-promoting activity from fetal bovine serum for human diploid fibroblasts has been separated from fetuin and serum albumin and purified approximately 15-fold.


Control of DNA synthesis in tissue culture cells. Eukaryotic DNA is functionally divided into thousands of replicons, each of which may be duplicated at a characteristic time within the DNA synthetic (S) period. Our approach toward an understanding of the molecular mechanisms which control orderly eukaryotic DNA synthesis has been: (a) to devise a method of cell synchrony in a suitable tissue culture system wherein all cells in the population enter and traverse the S period with a high degree of synchrony; (b) to determine, utilizing this system, precisely when during the S period critical events and macromolecular syntheses occur; and (c) to examine, by polyacrylamide-gel electrophoresis, the spectrum of proteins which become associated with chromatin during the S period in such a way as to suggest their involvement with DNA synthesis. Possible mechanisms for control are discussed based on the results presented here.


Human tolerance to impact acceleration.

Human tolerance to impact acceleration. Tolerance to brief impact is related to the imposed velocity change, whilst in longer impacts it is the level of acceleration which is critical. This behaviour results from the dynamic characteristics of the body which, for practical purposes, can be represented as a simple mass spring system. For any given acceleration vector, and for any given system of body restraint, whole body (primary) tolerance can be defined by two parameters--a critical velocity change and a critical plateau acceleration level. Appropriate figures are given for 14 different posture and restraint conditions. It is stressed that these are best estimates (or in some cases, best guesses) and that secondary or tertiary mechanisms of injury, in particular head strikes, must not be ignored in any real-life conditions of impact.


Constitutional factors and the local and general responses to injury. The different constitutional factors which may affect the response to injury and the problems which must be considered in evaluating any injury model are discussed. Species, age, sex, nutrition and the presence of disease are discussed in relation to an injury model based on hind limb ischaemia, with reference to the extensive literature.


Acute osteomyelitis following a closed fracture. Acute osteomyelitis following a closed fracture is very rare. A case following a closed epiphyseal (type II) injury of the distal end of the radius is reported and the role of injury as a contributing factor in acute haematogenous osteomyelitis is discussed.


Go-kart injuries.

Go-kart injuries. The pattern and number of injuries which have occurred over a period of 4 years at Aviemore Kart Tract are described. Some of the modifications which have significantly reduced the number and severity of the injuries are discussed.


Some aspects of post-traumatic elbow stiffness. In this article the anatomy of the soft tissues around the elbow joint is described in relation to the causation of post-traumatic elbow stiffness. Examples of the ways in which significant damage to these tissues may occur after injury are discussed. It is suggested how arthrography may help in the diagnosis of the condition.


The pattern of fractures and dislocations in western Nigeria. A consecutive series of 2579 patients with 2571 fractures and 189 dislocations was studied prospectively. Males and children, especially under 10 years of age, predominated. The majority of the injuries resulted from falls either in the home or outdoors and from road traffic accidents. Fractures occurred more commonly in the upper limb than in the lower limb or the axial skeleton. Fractures of the shaft of the femur were three times more common, while fractures of the neck of the femur were less common by the same factor, in Africans than in Caucasians. Dislocations were commonest at the shoulder and elbow. Only 11.4 per cent of the patients had open operations. The prognosis as regards survival and functional results was very good, but malunion was the most common complication.


Rupture of herniated small intestine from blunt abdominal injury: a report of 2 cases. Two cases of perforation of the small bowel following blunt abdominal injury in patients with an abdominal hernia are presented. In one case the bowel was present in a large incisional hernia and the injuring force caused transection of the bowel at three different sites. The bowel was resected and the hernia was repaired at the same time. The patient died of associated chest injuries. In the second case, the bowel was caught in an inguinal hernia; although the injuring force was relatively mild it resulted in two perforations in the herniated terminal ileum. The patient survived following closure of the perforations and simultaneous repair of the hernia.


The role of osteomedullography in the management of delayed union and non-union of diaphyseal fractures of the long bones: a preliminary report. The significance of osteomedullography was studied in 17 patients with diaphyseal fractures of long bones to determine a rational form of treatment for delayed healing and non-union. Positive osteomedullographic signs were noted in 6 cases of delayed healing and negative signs in 11 cases. All the positive cases united by prolonging the period of immobilization in plaster. Seven of the 11 negative cases were treated by bone grafting. Three patients treated by prolonged immobilization in plaster have not shown evidence of union even after a period of 20 weeks. This simple investigation could be useful in avoiding undue loss of time in the management of delayed union and non-union.


The use of external skeletal fixation in the treatment of fractures of the humeral shaft. A method of external fixation is presented which has been applied in 8 patients with a fracture of the humeral shaft. The indications and technique are described.


False aneurysm of the profunda femoris artery following internal fixation of an intertrochanteric femoral fracture. A case is presented of traumatic aneurysm of the profunda femoris artery following spontaneous unscrewing of the nut at the nail--plate junction of a two-piece nail plate which was used for internal fixation of an intertrochanteric femoral fracture. Methods of avoiding this complication during hip operations are described, together with a short review of the literature.


Synthesis of monorhamnosyl L-rhamno-D-mannans by conidia of Sporothrix schenckii. A rhamnomannan containing single-unit alpha-L-rhamnopyranose side chains was identified in isolated conidia from Sporothrix schenckii. Such a rhamnomannan differed from the dirhamnosyl rhamnomannan synthesized by the hyphae but was very similar to the monorhamnosyl rhamnomannan formed in yeastlike cells. Nuclear magnetic resonance spectroscopy and chemical analysis were used to compare these polysaccharides. Based on the distribution of different rhamnomannans in different S. schenckii cell types and in view of the reactivity of some human antisera previously reported (6), the formation of hyphae in vivo is suggested.


Quantitative measurements of cholera enterotoxin in cultures of toxinogenic wild-type and nontoxinogenic mutant strains of Vibrio cholerae by using a sensitive and specific reversed passive hemagglutination assay for cholera enerotoxin. A sensitive and specific reversed passive hemagglutination (RPHA) assay for cholera enterotoxin has been developed. Equine anti-choleragenoid antibodies purified by immunoadsorption were covalently coupled to formalinized sheep erythrocytes, using bis-diazotized benzidine, and the antitoxin-sensitized erythrocytes were shown to agglutinate specifically in the presence of cholera enterotoxin. In a microtiter RPHA assay system, the smallest quantity of enterotoxin that caused hemagglutination was approximately 20 pg. A sensitive assay for antibodies to enterotoxin was also developed, based on inhibition of RPHA. Using such assays, we demonstrated that several nontoxinogenic (tox-) strains of Vibrio cholerae produced small but detectable yields of enterotoxin, 4 to 16 ng/ml, under conditions where the highly toxinogenic strain 569B Inaba produced approximately 16 microgram of enterotoxin per ml. The enterotoxin produced in small quantities by these tox- strains was found to be identical to the enterotoxin from V. cholerae 569B Inaba iv its immunological and biological activities. Strains of V. cholerae that produce intermediate yields of enterotoxin have been obtained by two techniques: (i) as less toxinogenic mutants derived from highly toxinogenic strains and (ii) as more toxinogenic mutants derived from tox- strains. Thus, the yield of enterotoxin in cultures of V. cholerae grown under standardized conditions is is a genetically controlled trait that can be altered by mutation.


Production of extracellular and cell-associated glucosyltransferase activity by Streptococcus mutans during growth on various carbon sources. The production of extracellular and cell-associated glucosyltransferase activity by Streptococcus mutans strain GS-5 was examined during growth on various carbon sources in a chemically defined medium. S. mutans cells produced glucosyltransferase activity only during logarithmic growth when glucose, fructose, mannitol, or sorbitol was the sole carbon source. Cells growing on mannitol or sorbitol produced approximately half as much extracellular glucosyltransferase activity as cells growing on glucose, although the proportions of the glucosyltransferase activity capable of synthesizing insoluble glucans were similar. Cells growing on fructose produced slightly more extracellular glucosyltransferase activity than cells grown on glucose, yet the proportion of the glucosyltransferase activity capable of synthesizing insoluble glucans was again similar to glucose cultures. S. mutans cells growing in the presence of both glucose and mannitol displayed diauxic growth and initial preferential utilization of glucose. Glucosyltransferase enzyme production occurred only during the phases of cell growth in the presence of the two carbon sources. The cell-associated glucosyltransferase activities of glucose-, fructose-, mannitol-, and sorbitol-grown cells were relatively low, yet all the cells were capable of adherence to glass in the presence of sucrose. When glucose-containing cultures of S. mutans were supplemented with sucrose, extracellular glucosyltransferase activity first became cell associated and then appeared to become inactivated, presumably due to the accumulation of insoluble glucans.


Cell-mediated immune responses of guinea pigs to an inactivated phase I Coxiella burnetii vaccine. The ability of a killed phase I Coxiella burnetii vaccine to induce cell-mediated immune responses in guinea pigs was studied. Cell-mediated immune responses were assessed by the inhibition of macrophage migration and lymphocyte transformation assays. The macrophage migration response occurred rapidly and was detected at high levels, but was relatively short-lived. In contrast, the lymphocyte transformation response developed more slowly, and persisted for a longer period. The vaccine, given in a single dose or in two doses 1 week apart, protected guinea pigs from a subsequent virulent challenge.


Effective immunity to dental caries: dose-dependent studies of secretory immunity by oral administration of Streptococcus mutans to rats. Rats (COBS/CD) provided Formalin-killed Streptococcus mutans 6715, C211 in their drinking water (10(8) to 10(9) equivalent colony-forming units [CFU] per ml) had high levels of specific antibodies in saliva, colostrum, and milk. Rats provided a lower concentration of S. mutans antigen (10(7) CFU per ml) in water had agglutinin titers in secretions that were similar to those in controls. Gnotobiotic rats provided S. mutans antigen in food (10(7) to 10(8) equivalent CFU per g of diet) manifested a secretory immune response as evidenced by the presence of specific immunoglobulin A antibodies in saliva, colostrum, and milk. Gnotobiotic rats provided a higher concentration of antigen (10(9) CFU per g) in food had levels of specific antibodies in their secretions that were similar to those in controls. No significant antibody activity to S. mutans was observed in sera of any group of animals. Furthermore, the presence of specific salivary immunoglobulin A antibodies in gnotobiotic rats correlated with a reduction in the level of plaque, numbers of viable S. mutans in plaque, and levels of S. mutans-induced dental caries. This paper discusses the importance of antigen dosage for induction of a secretory immune response that is protective against S. mutans-induced dental caries.


Limits of rickettsial infectivity.

Limits of rickettsial infectivity. Seven species of pathogenic rickettsiae were compared in five assay systems for group, species, strain, and phase differences in infectivity. The species examined include Rickettsia prowazekii (Breinl and Cairo 3 strains), R. typhi, R. canada, R. rickettsii (Sheila Smith and R strains), R. conorii, R. sibirica, and Coxiella burnetii in phases I and II. Pathogenicity was charcterized in terms of fever in guinea pigs. All comparisons of infectivity and pathogenicity were described in terms of numbers of rickettsiae in the inocula, as determined by direct rickettsial count. The data characterize the various species and strains of rickettsiae in quantitative terms, which are also estimates of the sensitivity of the assay systems used. Phase I C. burnetii was found to be the most, and R. canada the least, infective of the species examined. In general the primary chicken embryo cell culture system proved to be the most, and that of the mouse the least, sensitive assay system.


Immunoelectron microscopic localization of lipopolysaccharides in the cell wall of Bacteroides oralis and Fusobacterium nucleatum. Lipopolysaccharides (LPS) have been extracted and purified from two anaerobic gram-negative bacteria: Bacteroides oralis and tfusobacterium nucleatum. Chemical analysis of the preparations showed a great proportion of neutral sugars, mainly glucose, in LPS of B. oralis. In rabbits, LPS of B. oralis induced both immunoglobulin M and G antibodies in contrast to LPS of F. nucleatum, to which only immunoglobulin M antibodies were produced. An immunohistochemical method with horseradish peroxidase-labeled antibodies was used to localize LPS antigens at the ultrastructural level. An electron-dense reaction product, representing an immune complex consisting of bacterial surface antigens and specific rabbit immunoglobulin labeled with peroxidase, was surrounding the cell wall, whereas appropriate controls were negative. The findings of the present study show that LPS of Bacteroides are probably bound to a complex, including glucans, in the outer membrane of the cell wall. LPS of Fusobacterium resemble LPS of other gram-negative bacteria.


Division of single host cells after infection with chlamydiae. Mouse fibroblasts (L cells) were infected in suspension with Chlamydia psittaci (6BC) and then plated out on a solid substrate at a density of 80 cells per cm2 so that the effect of chlamydial infection on the division of single host cells and their progeny could be determined. Uninfected L cells multiplied with a mean generation time of 15 h. The generation time of single L cells infected with 1.5 50% infectious units (ID50) of C. psittaci was over twice as long. Half of the infected L cells had divided once by day 4 after infection, and the rest had divided more than once. Division of infected cells frequently produced one infected and one uninfected daughter. About half of the L cells infected with 15 ID50 of C. psittaci divided at least once before most of them detached from their substrate before observation on day 3. Less than 10% of the L cells infected with 75 ID50 of C. psittaci divided before they were lost from their substrate by day 2. Comparable results were obtained with single L cells infected with a lymphogranuloma venereum (440L) strain of C. trachomatis and with single HeLa 229 cells infected with C. psittaci. It was concluded that high multiplicities of infection of host cells with chlamydiae quickly bring cell division to a halt, whereas lower multiplicities slow but do not immediately stop the division of host cells. However, indefinitely multiplying clones of chlamydia-infected host cells were not observed. The method used here should be applicable to other studies on the division of cells in culture.


Morphological features and functional properties of human fibroblasts exposed to Actinomyces viscosus substances. Connective tissue fibroblasts undergo cytopathic degenerative changes during certain long-term inflammatory diseases such as rheumatoid arthritis and periodontitis. The failure of inflamed tissues to repair properly may result from functional alterations of fibroblasts within the affected tissues. Numerous previous studies indicate that direct cytotoxicity by bacterial or other substances may be responsible for the cellular alterations observed in vivo. We have tested this hypothesis by exposing cultures of human diploid fibroblasts to homogenates of Actinomyces viscosus (a microorganism associated with periodontitis and capable of causing other chronic inflammatory diseases) and analyzing the effects on cell viability, morphology, and function. The cells bind and subsequently engulf relatively large quantities of the bacterial substances. These substances do not appear to be toxic to fibroblasts as determined by 51Cr release and microcytotoxicity assays, although there is a slight but significant decrease in protein synthesis (P less than 0.01) as measured by the incorporation of [14C]proline. However, collagen production was not altered, and the cytopathic alterations observed in diseased tissues in vivo did not occur in the exposed cells. These findings suggest that A. viscosus substances do not directly cause injury to connective tissue fibroblasts in periodontal disease but may, through cell-surface binding, mark these cells for subsequent immune-mediated damage.


Ammonium ion requirement for the cell cycle of Mycobacterium avium. Mycobacterium avium has a defined cell cycle in which small cells elongate to about five times their original length and then divide by fragmentation. The nitrogen requirement for production of maximal number of colony-forming units was assessed by varying concentrations and kinds of nitrogen source in the medium. Ferric ammonium citrate at a concentration in 7H10 medium of 0.17 mumol/ml or ammonium chloride at 0.25 mumol/ml as the nitrogen source permitted the cells to elongate and to undergo limited division, with the final culture at 4 x 10(7) colony-forming units per ml. Ammonium chloride at 2.5 mumol/ml or glutamine at 1.37 mumol/ml supported completion of the cell cycle with final colony-forming units at about 5 x 10(8)/ml. Other amino acids, including glutamic acid, at 2.5 mumol/ml did not support completion of the cell cycle, although in most cases an intermediate number of colony-forming units per milliliter were formed. Limited uptake of [(14)C]glutamic acid and uptake of [(14)C]glutamine were not detectable until cell fission began. Cells not limited for nitrogen took up five times as much (35)S during fission as limited cells did during the same time. The nonlimited cells contained 10 times as much sulfolipid as the nitrogen-limited cells at the end of the cell cycle. These results demonstrate that rapidly dividing cells of M. avium utilize amino acids and sulfur and also synthesize sulfolipids in events that are apparently separable from metabolic functions of elongating cells. The results are contrasted with those found for other mycobacteria in which no cell cycle has been demonstrated.


Identification of the virulence-associated antigen on the surface fibrils of Actinomyces viscosus T14. Actinomyces viscosus T14V is virulent (V) for monoinfected rats, causing periodontal disease and bone loss, whereas, A. viscosus T14AV, a mutant strain, is avirulent (AV). Surface antigens from the T14V and T14AV strains were prepared by lysozyme digestion of cell walls and were compared by immunodiffusion against antisera to T14V and T14AV whole cells. The V-associated antigen (V-antigen) was detected readily in the T14V, but not readily in the T14AV cell wall extract. Antiserum specific for the V-antigen was prepared by absorbing anti-A. viscosus T14V serum with cell walls from the T14AV strain. This antiserum was used in the indirect peroxidase-labeled antibody technique to localize the V-antigen on the bacterial cell surface at the ultrastructural level. With whole bacterial cells, the V-antigen was found on fine fibrils and was detected in both the T14V and T14AV strains. The presence of V-antigen on the AV strain was supported by the demonstration of antibodies against the V-antigen in anti-A. viscosus T14AV serum. Examination of isolated bacterial cell walls revealed a greater amount of fibrils and V-antigen on the T14V cell wall than on the T14AV cell wall. The data suggest that the presence of V-antigen represents a quantitative rather than a qualitative difference between the V and the AV strains of A. viscosus T14. Samples of human plaque were examined, and the V-antigen was found to be a specific marker for the fibril-containing layer of certain plaque bacteria, which are probably strains of A. viscosus or A. naeslundii.


Humoral and cell-mediated immune responses in humans before and after revaccination with vaccinia virus. Twenty-six healthy males vaccinated 15 to 18 years ago with vaccinia virus were revaccinated. Blood samples were collected before vaccination and 3 weeks after. The lymphocytes were tested in a blast transformation assay with vaccinia antigen and phytohemagglutinin, and interferon production was measured. The sera were subjected to neutralization and antibody-dependent cell-mediated cytotoxicity (ADCC) tests. All results were compared with clinical responses. The only test showing immunity in all donors before vaccination was the ADCC. The other tests showed a very limited residual immunity or no immunity at all. After revaccination, immunity reactions were found in all tests in most of the donors. None of the tests made before vaccination could be used to predict clinical reactions. The ADCC is recommended as a sensitive serological test.


Establishment of Marek's disease lymphoblastoid cell lines from transplantable versus primary lymphomas. Six new Marek's disease (MD) lymphoblastoid cell lines were established in vitro by cultivation in a medium containing 2-mercaptoethanol (2-ME). Attempts using primary lymphoma cells were generally unsuccessful; only one of 28 lymphomas yielded a cell line and that one came from an experimentally immunosuppressed chicken. In contrast, two of seven low-passage, and two of two established MD transplantable lymphomas grew readily in vitro. A sixth line was obtained using buffy coat cells from a leukemic chicken. It was concluded that the use of transplantable tumor cells and a medium containing 2-ME provided a combination highly suited to the establishment of cell lines from MD.


Effects of presensitization in vivo on cell-mediated responses to embryonic antigens in vitro. Lymphoid cells specifically reactive with antigens shared by rat bowel carcinomas and mid-term embryo cells were generated by in vitro culture on monolayers of embryo cells. Spleen cells from WF females were cultured for 5 days on monolayers of syngeneic embryo cells or adult cells and assayed for cytotoxic activity on syngeneic embryo, bowel carcinoma, or adult fibroblast target cells in microcytotoxicity and 51Cr-release assays. After culture on embryo monolayers, spleen cells were cytotoxic for embryo and tumor but not for adult fibroblast target cells. Enhanced cytotoxicity was recorded when the spleen cells were cultured from females after interstrain (WF X BN) pregnancy rather than from virgin females. In contrast, previous intrastrain (WF X WF) pregnancy appeared to depress the generation of spleen cells cytotoxic for target cells bearing embryonic antigens.


Characteristics of hamster cells transformed by the combined action of chemical and virus. Pretreatment of hamster cells with chemical carcinogen enhances transformation by a simian adenovirus, SA7. Such transformants were compared to cells transformed by SA7 alone for the presence of virus-specified antigens, and for the ability to clone in soft agar and to form progressively growing sarcomas in hamsters. The SA7 "T" antigen content and the relative cloning efficiencies in agar of cell lines of both groups were similar. Some cell lines obtained by pretreatment with chemical carcinogen, however, produced a higher incidence of tumors, suggesting that the chemical carcinogen was involved in the transformation of cells by virus. A further indication that these cells differed from cells transformed by virus alone was obtained with MCA-transformed fibroblast cell lines that were subsequently also transformed in vitro by SA7 to cuboidal (SA7 type) morphology. These MCA-SA7-transformed cells produced a higher  incidence of sarcomas than cells transformed by virus alone, but a lower tumor incidence than the parental MCA line.


BCG vaccination and the incidence of lymphomas and leukaemia. BCG vaccination has been claimed to prevent leukaemia, but there is also concern that it might increase the risk of Hodgkin's disease and other lymphomas. Both hypotheses were tested by comparing mortality and registration rates in cohorts of children from the North and South Islands of New Zealand. When school-children were offered vaccination in both islands, subsequent death rates from lymphomas and leukaemia were similar in the two islands. After withdrawal of vaccination in the South Island, mortality and registration rates for Hodgkin's disease remained similar in the two islands, but there was significant excess of deaths from non-Hodgkin lymphomas in the North Island. The difference in the registration rates for these tumours was much smaller than the difference in mortality rates and was not statistically significant. There was no evidence that BCG vaccination in the North Island prevented leukemia. These findings and the results of other studies suggest that proposals for BCG vaccination against leukaemia are unwise.


Inhibition and promotion of tumor growth by BCG: evidence for stimulation of humoral enhancing factors by BCG. Effects of pretreatment with BCG, strain Japan, on tumor growth were studied using a transplatable methylcholanthrene (MCA)-induced fibrosarcoma in C3H/He mice. Injection of BCG7 weeks before tumor inoculation at a site distant from the tumor caused a slight inhibition of tumor growth. A low dose of tumor cells did not grow at the BCG-primed site when BCG was injected 7 and 11 weeks before the tumor. When a high dose was inoculated into the BCG-primed site, inhibition of the primary tumor occurred in mice which had received BCG 7 weeks previously, but the number of distant metastases in the popliteal lymph node and the lungs was increased in mice pretreated with BCG at any time. Furthermore, post treatment with BCG at a site distant from the tumor caused promotion of tumor growth. Enhanced antibody formation and suppression of delayed type hypersensitivity (DTH) occurred in tumor-bearing mice. BCG treatment of such mice caused a vigorously enhanced antibody formation and a marked suppression of DTH. The sera from tumor-bearing mice enhanced tumor growth. Tumor growth was suppressed in splenectomized mice. These findings suggested that antibodies against tumor-specific antigens enhanced tumor growth in this system and that BCG treatment of tumor-bearing mice stimulated formation of antibodies probably acting as blocking factors.


Relationship between clearance of lithium, potassium and sodium in human mixed saliva. Samples of saliva were collected from healthy subjects given a test dose of lithium carbonate. Lithium clearance in saliva was well related to salivary potassium clearance but not to salivary sodium clearance. Stimulation of adrenocortical activity by changing from the recumbent to the upright posture caused salivary sodium clearance to fall, but failed to affect the clearance of lithium and potassium in saliva. The findings show that the secretion of lithium in saliva resembles potassium secretion more than sodium secretion.


Use of psychotropics in the world.

Use of psychotropics in the world. A questionnaire regarding medication preferences for major categories of psychiatric disorders was sent to 1,059 psychiatric drug investigators in 53 countries. 264 questionnaires were returned, of which 165 were appropriate for this survey. A total of 87 different psychotropic drugs were selected. Chlorpromazine was the medication most frequently cited in the treatment of schizophrenia. Amitriptyline and imipramine together accounted for the vast majority of medication chosen for all varieties of depression. In the treatment of mania, chlorpromazine was chosen by almost one-third of our sample, lithium by only one-fifth. Chlordiazepoxide and diazepam were equally preferred in the treatment of alcoholism. Most psychiatrists preferred not to use any psychotropic medications consistently in treating patients with organic brain syndromes. The implications of this study are discussed and compared uith similar studies in more limited geographical regions and in children.


The daily rhythm of shedding and degradation of rod and cone outer segment membranes in the chick retina. Newly hatched chickens were maintained on a daily light cycle of 12 hr of light and 12 hr of darkness for 12 days. The pigment epithelium was then examined by electron microscopy at different times of day. Shortly after the beginning of the light period, the rods discarded groups of outer segment membranes. During the remainder of the light period, the membranes were degraded by the pigment epithelium. Early in the dark period, the cones shed membranes, which were digested by the pigment epithelial cells during the subsequent hours of darkness. Available evidence suggests that at least some of the chemical activities of the visual cells and pigment epithelium oscillate with a daily rhythm, which is synchronized with the daily fluctuation of light in the environment.


Functional competence of regenerating ocular surface epithelium. Several aspects of the ocular surface epithelium, including its clinical appearance, its effect on the strength of underlying stromal wounds, and its healing rates, have been compared between regenerating epithelium of conjunctival and that of corneal origin in rabbits. The results showed that conjunctival epithelium could not transform completely into functionally competent corneal epithelium within 6 weeks.


Studies on the crystalline lens. XXVI. Kinetic study showing saturation of the sodium pump. Active transport of sodium out of rabbit lenses was shown to be carrier mediated by observing the dependence of efflux of 22Na on intracellular concentration of nonlabeled sodium which previously was elevated by incubating lenses at 0 degree C. Velocity of sodium efflux increased asymptotically with increasing concentrations of intracellular sodium, demonstrating saturation of the carrier. The maximum velocity of the sodium pump based on Michaelis-Menten kinetics was found to be 4.6 mumol/hr/lens, and the half-saturation of the carrier, which appeared to involve a single site, occurred at 36 mM.


Fine structure and radioautography of rabbit photoreceptor cells. The retina of the adult rabbit contains two types of photoreceptors. Type I predominates and is a typical rod. The identity of the type II cell has been uncertain until now. This study demonstrates that the type II cell has certain conelike morphologic features including a pale nucleus, complex synaptic pedicel, and multiple wrappings of the outer segment by microvilli of the pigment epithelium. The type II outer segment is also conelike in that some intradisc spaces are patent to the extracellular space, that the pattern of renewal is diffuse as demonstrated radioautographically with sugar and amino acids as precursors, and that the few examples of distal tip shedding found in this study occur mainly in the early part of the dark cycle. The interior of the type II outer segment is labeled selectively with 3H-fucose, whereas the interior of the type I outer segment remains unlabeled with this sugar at all time intervals examined. The label probably reflects the presence of a fucose-containing glycoprotein in the conelike outer segment which is lacking in the rod outer segment and may represent a visual pigment or other intrinsic molecule(s). An amorphous material which forms a symmetric sleeve around the photoreceptor connecting cilium has been demonstrated after intravitreal injections of horeseradish peroxidase. This cilium-associated material is labeled specifically by 4 hr following intravitreal injection of 3H-fucose, before the interphotoreceptor matrix shows significant labeling, suggesting that it is composed of a sugar-containing material. Possible functions of the cilium-associated material, including structural support and ionic regulation, are discussed.


[Prognosis with different therapeutic methods and therapeutic recommendations in operable breast cancers]. Breast carcinoma is the most frequent form of cancer in women. In Switzerland 1265 women died of it in 1975, the mortality rate is rising slowly, the cause of this fact being unclear. There is no uniformity about the best treatment of breast cancer. We analyzed 454 carcinomas, operated in the years 1961--1971 after Halsted, Patey and McWhirter. We found similar 5- and 10-year survival rates with the Halsted and the Patey method of operation, however, decidedly worse results with the simple mastectomy. Nowadays we recommend the modified radical excision of the breast after Patey at all stages; in addition we treat all patients with postoperative X-ray therapy or prescribe an adjuvant cytostatic therapy based on the experimental study of the SAKK (Swiss Therapy Group for Clinical Cancer Research).


[Mammographic and thermographic diagnosis of breast cancer]. 2731 mammography and thermography results were analyzed. The radiological examination could be histologically verified 468 times. The diagnosis made by means of mammography was wrong in 42%, by thermography in 52%. In this group only 3 occult carcinomas were discovered mammographically. A wide range of indications of mammography applied hitherto therefore becomes doubtful. It is recommended to assign all palpable tumours of the female breast directly to the surgeon without mammography.


[Indication for subcutaneous mastectomy as carcinoma prevention]. The indication for subcutaneous mastectomy is analyzed. Prospective investigations seem to confirm that different morphological forms of breast dysplasia (grade I, II, III, PRECHTEL [10]) do not implicate differences in risk for development of invasive breast cancer. Only if these forms of breast dysplasia are associated to high risk factors such as family history of breast cancer, should subcutaneous mastectomy be considered as a cancer prophylaxis.


[Surgical treatment of pulmonary coin lesions found in cancer patients and 6 breast cancers. Solitary metastasis? 2d primary bronchopulmonary cancer? Coin lesions of benign nature?]. Pulmonary diffusion of breast cancer is frequent in those patients who have died of the disease and in those survivors who have not been cured after removal of the breast and X-ray treatment in the advanced states of the disease. When the metastases are identified, they are almost always multiple and bilateral. The appearance of a solitary, late, pulmonary coin lesion (metachrone), in someone with breast cancer certainly suggests a pulmonary metastase, but in fact, it is more likely to be a second cancer than a metastase, that is, a primary bronchopulmonary cancer. The presence of a solitary pulmonary coin lesion in someone who has or who has had breast cancer, presents therefore certain particular problems. After having controlled by xerotomography that there is no pulmonary diffusion in either lung, that there is no invasion of other tissues or organs, and after having controlled locally around the breast cancer, then it is imperative to remove the lesion without delay since it is certainly malignant and most probably a second cancer, that is, a primary bronchopulmonary cancer, an adenocarcinome, detected at an asymptomatic stage. The prognosis of a broncho-pulmonary adenocarcinome depends on the precocity of its removal.


[Role of surgery in the treatment of breast cancer in surgical border-line cases]. Since the turn of the century, a selection of borderline operable cancers of the breast have benefited of preoperative radiotherapy. Adjuvant chemotherapy may enlarge the limits of operability in a foreseeable future especially if applied before surgery. Different clinical classifications and staging procedures are studied as well as their influence on prognosis and their use in evaluating the limits of operability.


[Indications and procedures in conservative goiter treatment]. A review of the literature shows that thyroid hormone therapy causes 13 to 40% of euthyroid goiters to disappear and brings about a reduction in goiter size in another 30 to 50%. The hormonal treatment is more successful with smaller goiters. Uninodular goiters respond on the average less well than diffuse and multinodular goiters. In Switzerland, about 33% of adult persons have goiters, albeit mostly of small size. This renders impracticable investigation or treatment of all goiters. It is recommended that only goiters which are esthetically disturbing or which cause pressure symptoms be treated. An operation must always be performed when there is suspicion of thyroid cancer or in the presence of threatening pressure symptoms. In all the other cases, thyroid hormone therapy may be tried. If an operation cannot be performed for cardiac or other reasons, radioiodine will relieve pressure symptoms due to goiter in over 65% of cases.


[Surgery of benign goiter].

[Surgery of benign goiter]. All our thyroid patients are seen jointly by a specialist in nuclear medicine and by an endocrinologist. From 1972 to 1976 we operated on 564 patients; 433 were benign goitres (152 of them hyperthyroid) and 131 were malignant thyroid tumors. Operations for recurrent benign goitre were performed on 61 patients. The frequency of unilateral recurrent laryngeal nerve paralysis following a first operation (checked by laryngoscopy) was 4,4%. The surgical measures for avoiding this complication are discussed.


[Surgical treatment of malignant thyroid neoplasms]. The classification of thyroid tumors, based on morphological and clinical aspects, makes it possible to compare the experience of several centers. The histology of the tumor and the age of the patient are the main criteria of prognosis. A differentiated tumor may become anaplastic. The therapeutic approach, which is primarily surgical, is therefore different for differentiated, medullary and anaplastic carcinomas. In the field of differentiated tumors, there are still some differences of opinion regarding the radicality of thyroidectomy and node dissection. In planning surgery, the incidence of bilateral tumor according to histological type, the pattern of primary lymphatic drainage and the risk of parathyroid insufficiency should be taken into account. The collaboration of the pathologist is essential. The results should be appreciated after a follow-up of at least 10 years.


[Pathophysiology and surgery of the parathyroid glands]. The treatment of primary hyperparathyroidism consists in a teamwork between the physician, pathologist and surgeon. The endocrinologist must be able to motivate the surgeon to perform an often tedious exploration of the neck, while the pathologist may influence the tactics of surgery by his analysis of the frozen sections. The surgeon needs much experience, a sound knowledge of anatomy, a good surgical technique and enough time and leisure for the operation.


[Reconstructive surgery of the peripheral arteries in elderly people]. The results of reconstructive surgery of atherosclerotic occlusive disease in the senium are reported on 71 patients. Mortality rate of 14.68% and improvement of 63.38% showed clearly better results than described in literature with an amputation mortality rate between 26% and 60%. In the authors' opinion also in the senium the reconstructive surgery of the peripheral arterial system should be preferred if general and local operability are given.


[Wound infections in arterial surgery of the lower extremities]. A survey showed that in 764 cases of reconstructive vascular operations of the lower extremities there were 5.14% bland infections, 1.14% were, however, severe. In 78.5% of these infections synthetic materials were used. Autologous venous tissue for bypass or patches showed the lowest risks. In the case of postoperative signs of an infection on an arterial suture, a secondary bypass well in an area of healthy tissue should immediately be attempted to save the limb.


[Could the outcome of therapy of deep thrombophlebitis be improved with a combination of surgical thrombectomy and regional fibrinolysis?]. A new mode of treatment of extensive acute and subacute deep venous thrombosis of the lower extremities is introduced. For this purpose the beneficial effects of surgical thrombectomy and of thrombolysis with streptokinase are combined during the course of a single surgical intervention. Rapid-flow regional perfusion is the vehicle used for administration of streptokinase and probably represents the third arm of this therapeutic approach by adding a hemodynamic wash-out effect. Because the thrombolytic agent is rinsed out of the circuit at the end of regional perfusion the usual side effects and contra-indications of this drug are avoided. Early and late results of this treatment are assessed clinically and with repeat venograms in a group of six unselected patients. Highly satisfactory results were obtained in four patients with complete anatomical and functional restoration of deep veins along their entire length in three cases. It is felt that continued use of this method is warranted.


[The risk of lung emolism in isolated thrombosis of the V. saphena magna]. Within 1 1/2 years we have seen 4 pulmonary emboli, one of which was fatal. The origin was an isolated thrombophlebitis of the great saphenous vein. An autopsy, with dissection of the veins of the legs and of the pelvis, and phlebographies where there is any doubt of deep venous thrombosis, allow us to speak of isolated thrombosis of the great saphenous vein. The clinical level is always lower than the real level and the thrombus can grow through the saphenofemoral junction. The conservative treatment (anticoagulation and mobilisation) do not prevent a cranial ascension of the thrombus nor a pulmonary embolism. So, when the thrombosis is higher than the knee, we do a crossectomy of the great saphenous vein. We did this operation 8 times without complications.


[Bovine graft as arteriovenous fistula for hemodialysis]. We used the bovine carotid artery (BCA) as arteriovenous shunt for hemodialysis in 11 patients since December 1975. We discuss our results. For implantations we used two methods: straight and loop grafts. The BCA have many advantages as: good access for punction, anticoagulation not permanently necessary, volume of shunt more than 250 ml/min; the disadvantages are: increased tendence to infection and thrombosis. Thrombosis in BCA can be easier removed than in other grafts. Reduction of the complications incidence is possible with an accurate technique. We think that the BCA is very useful when Cimino-Brescia fistula has failed or is unavailable; we prefer BCA to a saphenous arteriovenous graft in these situations.


[Experiences with percutaneous K-wire osteosynthesis in distal radius fractures]. Follow-up controls were made on 300 patients whose fractures of the distal radius were treated by a primary operative fixation with percutaneous K-wire. This group showed significantly better results than a control group of 300 patients given conservative management. We recommend a primary percutaneous K-wire osteosynthesis as treatment of choice for the management of distal radius fractures.


Malignant hyperthermia in a Greyhound.

Malignant hyperthermia in a Greyhound. A rapid rise in rectal temperature (to 45 C) in a young male Greyhound was attributed to the use of halothane and succinylcholine. Circulatory failure and death occurred within approximately 90 minutes of the onset of the hyperthermia.


Retrospective study of an Ehrlichia canis epizootic around Phoenix, Arizona. Antibodies to Ehrlichia canis were detected by indirect immunofluorescence in sera from 40 of 339 dogs (12%) examined from around Phoenix, Az, during 1975. The prevalence was higher in purebred and crossbred German Shepherd Dogs (21%), as compared with 8% in the remaining breeds. The difference in antibody prevalence between sexes was not significant. Dogs 12 weeks to 13 years old were seropositive. The causative agent was isolated by inoculation of whole blood from a seropositive dog into a susceptible German Shepherd Dog.


Survey of conjunctival flora in dogs with clinical signs of external eye disease. Bacteria were isolated 68% of the time from the conjunctival sacs of 120 dogs with conjunctivitis, blepharitis, dacryocystitis, or corneal ulcerations. Coagulase-positive Staphylococcus aureus was the most frequently isolated organism (68%); when both eyes were affected, S aureus was recovered from 29% of the dogs; and in dogs with 1 eye affected, S aureus was isolated from 23% of the affected eyes and in 18% of the nonaffected eyes. Other organisms recovered from eyes were coagulase-negative Staphylococcus epidermidis (27%), beta-hemolytic streptococcus (19%), alpha-hemolytic Streptococcus (17%), Escherichia coli (10%), Bacillus spp (11%), and Proteus spp (7%). It was concluded that S aureus may appear as a primary pathogen in ocular disease. When 1 eye is clinically affected, S aureus can be a potential invader of the normal eye.


Renal failure and hyperparathyroidism in an Alaskan Malamute pup. In a 5-month-old male Alaskan Malamute, clinicopathologic and radiographic findings supported a diagnosis of renal failure and hyperparathyroidism. At necropsy, the parathyroid glands were enlarged and the kidneys were shrunken and irregular. The histopathologic findings were consistent with the diagnosis of logic findings were consistent with the diagnosis of terminal renal failure and hyperparathyroidism. Based on the history, a heritable disorder was suspected.


Isolation of Treponema hyodysenteriae from sources other than swine. Fecal samples were collected from animals and environments on 3 swine farms and cultured for Treponema hyodysenteriae. Each farm was a farrow-to-finish operation and, at the time of sampling, swine dysentery was enzootic among 8- to 22-week-old pigs. Pathogenic T hyodysenteriae was isolated from pigs on all 3 farms. On farm A, nonpathogenic T hyodysenteriae was isolated from a sample of lagoon water. On farm B, pathogenic T hyodysenteriae was isolated from a waste-holding pit. On farm C, a dog was observed to be eating feces of pigs that had swine dysentery. The dog was diarrheic and a fecal sample yielded a pathogenic isolant of T hyodysenteriae. Further isolation attempts were unsuccessful after the dog was removed from the infected premises. Isolation of pathogenic and nonpathogenic organisms from waste-holding systems emphasizes the need for cultural techniques in detecting pathogenic T hyodysenteriae.


Swine dysentery: studies of gnotobiotic pigs inoculated with Treponema hyodysenteriae, Bacteroides vulgatus, and Fusobacterium necrophorum. Transmission experiments were carried out in gnotobiotic pigs to determine whether lesions typical of swine dysentery could be produced by oral inoculation of Treponema hyodysenteriae in combination with Bacteroides vulgatus or Fusobacterium necrophorum, or both. Each of the organisms had been isolated from swine with early lesions of the disease. Lesions were not found in 6 pigs inoculated with T hyodysenteriae alone, in 4 pigs given F necrophorum and T hyodysenteriae, or in 4 pigs given B vulgatus and F necrophorum. Lesions typical of swine dysentery developed in 8 pigs given B vulgatus, F necrophorum, and T hyodysenteriae as well as in 3 of 4 pigs given B vulgatus and T hyodysenteriae. In both of these groups, the inoculated bacteria were recovered from the colon, and T hyodysenteriae was demonstrated in the colonic crypts, epithelium, and lamina propria. The pathogenicity of the T hyodysenteriae was shown by the development of characteristic signs and lesions of swine dysentery in 12 of 14 naturally farrowed pigs inoculated with T hyodysenteriae alone.


Mycoplasma alkalescens-induced arthritis in dairy calves. Mycoplasma alkalescens was isolated from 6 of 7 synovial fluid samples taken by arthrocentesis from 3-week- to 4-month-old Holstein-Friesian calves with severe arthritis (tibiotarsal or carpal joints). Approximately 30 of 215 calves in the herd were affected. In one 6-week-old calf, M alkalescens was isolated from the liver, right tibiotarsal joint, right and left popliteal lymph nodes, and an exposed umbilical artery. Intraarticular inoculations of broth cultures of M alkalescens initially induced a febrile response and then severe fibrinopurulent arthritis. Intravenous inoculation of M alkalescens induced only a febrile response. The natural disease may have been a complication of umbilical exposure to M alkalescens, causing omphaloarteritis and subsequent arthritis. Before and during the arthritis problem, the umbilicus of newborn calves was dipped in an organic iodine product with 10% glycerin, marketed as a postmilking teat dip. After the cause of the arthritis was determined, the umbilicus of each newborn calf was treated with 7% tincture of iodine and no new cases of arthritis occurred.


Postpartum evaluation of bovine reproductive potential: comparison of findings from genital tract examination per rectum, uterine culture, and endometrial biopsy. The genital tract of 106 cows was evaluated after parturition by rectal palpation and by visual inspection of vaginal discharges. Analysis of these results revealed a significant (P less than 0.01) correlation with the ratings of endometrial biopsies. The most frequent bacterial isolations included Corynebacterium pyogenes, coliforms, or streptococci, or combinations thereof. Abnormal ratings obtained by gross evaluation and by histologic examination were associated principally with C pyogenes. Purulent discharges in particular were highly correlated (P less than 0.01) with infection caused by C pyogenes. Neither the gross genital tract condition nor the uterine histopathology rating was positively correlated with conception rate; however, the association between days open and conception rate was significant (P less than 0.05). Bacterial infections significantly (P less than 0.05) increased the number of services required for conception and significantly (P less than 0.01) increased the number of cows culled because of infertility. Cows up to 4 years old had a 61% 1st-service conception rate compared with 35% for older cows, and this difference was significant (P less than 0.05). The development of one or more postpartum diseases significantly (P less than 0.05) reduced 1st-service conception rate, whereas milk production was negatively correlated (P less than 0.05) with conception.


Tetany associated with magnesium deficiency in suckling beef calves. Sudden death losses occurred in 4-month-old beef calves with clinical signs of muscular dysfunction, including tetanic spasms. Gross lesions were not diagnostic. Microscopic lesions consisted of deposition of calcium salts in the elastic fibers of the arteries in the heart, lungs, and spleen and in the trabeculae of the spleen. Focal areas of interstitial nephritis were characterized by infiltration of lymphocytes and plasma cells. Samples of hay and silage that the cattle were consuming were low in magnesium. Calves with spasms responded to magnesium salts given intravenously, and losses ceased after a magnesium-containing supplement was fed free choice to the calves and cows.


Lead poisoning in feeder calves.

Lead poisoning in feeder calves. A group of 90 feeder calves was accidentally exposed to lead for approximately 30 days. The source of the intoxication was determined to be contamination of feed ingredients from a railroad car. Fourteen calves died and 8 more were clinically affected. Blood samples were obtained from 24 exposed calves (16 clinically normal and 8 intoxicated), and the samples were analyzed for lead, delta-amino levulinic acid dehydrase activity, and free erythrocyte porphyrin. Blood lead values ranged from 0.44 to 1.16 parts per million. Amino levulinic acid dehydrase activity was not affected enough to be of diagnostic value, whereas free erythrocyte porphyrin was increased dramatically and consistently in lead-exposed cattle.


Pulmonary arteries of the normal rat: the thick walled oblique muscle segment. The structure and certain metrical features of the pulmonary arterial tree in the normal rat have been studied in detail by both light and electron microscopy after its distension by injection. In the parts of the tree where the muscle wall is complete a segment with oblique muscle has been identified. The wall of this segment is thick, and is composed largely of obliquely orientated, closely packed smooth muscle cells with little intervening connective tissue, while it lacks a complete external elastic lamina. Both the axial pathway and its side branches have such oblique segments. Proximal and distal to an oblique segment there are transitional regions where obliquely arranged muscle fibres spiral external to the circularly arranged fibres usually described in muscular arteries. The existence of thick walled oblique muscle segments in the pulmonary arterial tree of the normal rat is important since this animal is frequently used in experimental studies, and the thick segments could be taken as evidence of induced hypertrophy.


Cell specialization in the small intestinal epithelium of adult Xenopus laevis: functional aspects. The functional activity of the epithelial cells of the intestinal folds of Xenopus was analysed histochemically. There is a gradient of increasing absorptive, digestive and secretory activity from the trough regions to the crests of the longitudinal folds. The differences in functional activity correspond to differences in structural complexity, and confirm the existence of a more specialized population of cells at the fold crests and a less specialized population at the fold troughs, analogous to mammalian villous and crypt cells respectively.


The growth of femur and tibia in three genetically distinct chondrodystrophic mutants of the house mouse. Growth of femora and tibiae has been measured in mice carrying three distinct chondrodystrophic mutants (achondroplasia cn, brachymorphic bm and stumpy stm) aged 6--128 days, and in normal litter-mates. cn and bm resemble each other in growing slowly until the time of weaning, when growth is interrupted, stm grows strongly at first, but stops at around 14 days. The significance of these findings is discussed.


Development of the indusium griseum. I. A quantitative light microscopic study of neurons and glia. A quantitative study of the development of the indusium griseum was carried out in mice. It was not possible to separate the indusium griseum from the cingulate cortex until birth. There was no increase in neuronal numbers between birth and 140 days postnatum, but glial cells probably increased in number from birth to 17 days; 70% of the adult total of glial cells were present at birth. The length of the indusium griseum increased from 1114+/-151 micrometer at birth to 2444+/-54micrometer at 25 days and 2544+/-80 micrometer at 140 days. Although the mean number of cells in equivalent sections decreased with age the distribution of cells was similar at all ages, with the majority of cells lying in the rostral half of the indusium griseum.


Sutural closure in rabbit and man: a morphological and histochemical study. Some aspects of normal obliteration of sutures in rabbit and man were studied histologically and it was found that suture closure took place by intramembranous ossification in specific areas of trans-suturally arranged, tendon-like tissue. Histochemically these areas demonstrated high activity of oxidative enzymes.


A maturation change in the surface of cat articular cartilage detected by the scanning electron microscope. The articular surface of the femoral condyle of kittens (2 days to 2 months old) and cats (6 months to 20 months old) was studied by examining air dried and critical point dried specimens in the scanning electron microscope. The surface of kitten cartilage was found to be populated by innumerable humps which were more prominent in air dried than in critical point dried specimens. Undulations and ridges were seen on air dried kitten cartilage, which was markedly shrunken and distorted, but undulations and ridges were absent from critical point dried specimens where shrinkage was more modest or even undetectable. The surface of the articular cartilage of 12 months and 20 months old cats was populated by innumerable pits. A rare hump in a pit was seen in specimens from the 12 months old cat, but not from the 20 months old animal. In 6--8 months old cats an intermediate situation prevails, for in some specimens pits were present on the posterior part of the condyle but humps and humps in pits were present on the anterior aspect. This study shows that the surface of young articular cartilage is populated by humps, but as the cartilage matures these formations are either transformed into pits or replaced by pits.


A quantitative light microscopic study of the indusium griseum of the mouse brain following hypoxic stress. A quantitative study of the indusium griseum was carried out in control mice and in mice subjected to hypoxia at 390 mmHg for 2 days. One group of mice subjected to hypoxia was killed immediately after hypoxia while another group was allowed to recover for 1 week. There was no significant variation in the number of neurons in any of the groups. Neuroglia decreased from a mean number of 1741 +/- 120 in the control animals to 1251 +/- 67 in the hypoxic group and 1223 +/- 51 in the recovery group. It is postulated that this fall occurs mainly in astrocytes because these cells appear to be especially susceptible to the effects of hypoxia.


Localization of the phrenic nucleus in the spinal cord of the rabbit. The motor fibres of the phrenic nerve arise in the cervical portion of the spinal cord from a group of lower motor neuron cell bodies called the 'phrenic nucleus'. The phrenic nucleus of the rabbit has been located by cutting the right phrenic nerve (phrenicotomy) in the neck and observing retrograde degeneration (chromatolysis) in the cells of the ventral grey horn of the spinal cord 2-4 weeks later. The phrenic nucleus in the rabbit was found to extend from the fourth to the sixth cervical segment. It lay in the caudal half to three fourths of the fourth, the whole of the fifth and the cranial half to two thirds of the sixth segment. The phrenic nucleus in the rabbit is a longitudinal column of cells lying between the ventromedial and the ventrolateral columns of the ventral grey horn of the spinal cord. In the fourth and fifth cervical segments, the phrenic nucleus of the rabbit is dorso-medial to the ventrolateral column and dorsolateral to the ventromedial column. In the sixth segment it is dorsomedial to the ventrolateral column but lateral to the ventromedial column. The total number of chromatolysed cells found in the right phrenic nucleus (after right phrenicotomy in the neck) varied between 1496 and 1754. The chromatolysed cells had a peculiar and characteristic orientation in that in a frontal section they appeared longer than broad with their long axes almost parallel ti the longitudinal axis of the spinal cord.


Epithelial mitotic activity in experimental gastric wounds. Excision wounds were made in the pyloric epithelium of the rat's stomach and the mitotic response studied at intervals from 1 day to 12 weeks afterwards. Increased mitotic activity followed wounding with peaks at 2 and 7 days, and with a subsequent high level of activity which was maintained in all the later stages of healing, even in the presence of long-delayed epithelialization. Most mitoses initially occurred some distance from the wound edge, but subsequently the highest counts were found in glands immediately adjacent to the edge, and this distribution persisted. Mitotic counts for deep ulcers (including mucous membrane and muscularis externa) were higher than those for superficial ulcers (involving mucous membrane only) at any one time.


Functional anatomy of the lungs of the green lizard, Lacerta viridis. The gas-exchange area in the lung of Lacerta viridis has been studied by light microscopy and electron microscopy. The interior of the lung in this species is partitioned into air sacs by radially disposed septa. The surfaces of each septum are covered by a continuous epithelium, the cells of which are termed 'pneumonocytes'. Deep to the epithelium there is a close-meshed plexus of capillaries. The middle layer of the septum contains smooth muscle and fibrous tissue. Two varieties of pneumonocytes can be identified. The type I cells are squamous and give off attenuated sheets of cytoplasm which spread widely over the septal surface; these sheets contain few organelles. The type II cells are more compact and possess many organelles; their osmiophilic inclusion bodies are especially conspicuous. The pulmonary capillaries of Lacerta are evaginated into the air sacs and often display marked attenuation of their endothelium. The possible functional significance of these features is discussed.


Amino acid sequences of the two polypeptide chains in beta1-bungarotoxin from the venom of Bungarus multicinctus. The two dissimilar composite polypeptide chains (A and B) in beta1-bungarotoxin were isolated as their reduced and carboxymethylated derivatives as reported in the preceding paper. The N-terminal sequences were determined with a sequenator up to the 39th residue for the RCM-A chain and up to the 25th residue for the RCM-B chain with repetitive yields of 90-95%. The tryptic and chymotryptic peptides from the two chains were isolated and their structures were determined by manual Edman degradation together with dansyl-Edman and carboxypeptidases A and Y. To complete the primary structures of the two chains, information on the tryptic peptides derived from the maleylated derivatives of the two chains was also used. The completed amino acid sequence of the A chain containing 120 residues (molecular weight, 13,500) is similar to that of notexin, a presynaptic neurotoxin from Australian tiger snake venom, and phospholipases A from other snake venoms. The amino acid sequence of the 60 residues in the B chain (molecular weight, 7,000) bears no resemblance to any basic polypeptides from snake venoms. The B chain probably plays a significant role by interacting with some components in presynaptic membranes of neurosmuscular junctions.


A study on nitrate reductase from Propionibacterium acidi-propionici. Cell extract from a strain of Propionibacterium acidi-propionici with high nitrate reductase (NaR) activity catalyzed nitrate reduction with glycerol phosphate, NADH, or lactate. The reaction was inhibited partially by fumarate or oxygen. NaR linked to methyl viologen was found mostly in particulate fractions. It was solubilized by treatment with Emulgen 810 and purified 46-fold by DEAE-cellulose, Sepharose 4B, and triple DEAE-Sephadex chromatographies in the presence of the detergent. It was rather labile but was stabilized by glycerol. The molecular weight was estimated to be 230,000 by Sepharose 4B gel filtration and the isoelectric point was pH 5.0-5.5. The pH optimum was at 6.5-7.5 and Km for nitrate was 0.1 mM. As electron donors, methyl and benzyl viologen were utilized well but FAD and FMN were fairly ineffective. Chlorate was an active acceptor as well as nitrate. Azide, cyanide, and thiocyanate inhibited NaR. On adding 1 mM tungstate to the growing medium, the NaR level in grown cells was lowered; addition of 0.01 mM molybdate restored the activity partially. NaR is suggested to be a molybdo-protein, similar to this enzyme from other bacteria.


Mode of inhibition of ornithine aminotransferase by L-canaline. The mechanism of inhibition of ornithine aminotransferase [EC 2.6.1.13] by L-canaline (alpha-amino-gamma-amino-oxybutyric acid) was investigated. Spectral changes of pyridoxal 5'-phosphate in ornithine aminotransferase on addition of L-canaline showed that L-canaline formed an oxime-type compound with pyridoxal 5'-phosphate that had the same spectra as the compound formed on addition of hydroxylamine to the holoenzyme. Kinetic studies indicated that hydroxylamine was a reversible noncompetitive inhibitor, whereas L-canaline was an irreversible inhibitor of ornithine aminotransferase. Other analogs, such as delta-aminovaleric acid and alpha-N-acetyl-L-ornithine, also reacted with the pyridoxal 5'-phosphate of the enzyme, but these compounds were competitive inhibitors with respect to L-ornithine. L-Canaline and hydroxylamine also reacted with pyridoxal 5'-phosphate in pig heart aspartate aminotransferase [EC 2.6.1.1] to produce an oxime, but both of them were reversible and noncompetitive inhibitors of the enzyme. The Ki value of hydroxylamine for ornithine aminotransferase was 4.3 X 10(-7) M and those of L-canaline and hydroxylamine for aspartate aminotransferase were 1.7 X 10(-4) M and 2.2 X 10(-5) M, respectively.


Circadian rhythm in the ornithine decarboxylase activity of rat small intestine. Intestinal ornithine decarboxylase [L-ornithine carboxy-lyase, EC 4.1.1.17] activity was found to show a marked circadian rhythm with a peak 4 h after the start of eating in rats on a diet containing protein. In rats with an intestinal blind loop, the enzyme was induced in the portion of the intestine that came in contact with the protein meal, but not in the blind loop. Injection of tetragastrin or CCK-PZ alone had no effect on enzyme induction, but when a protein suspension was introduced into a tied loop of intestine soon after the injection of tetragastrin or CCK-PZ, the enzyme was induced in the segment to almost the same extent as in the intestines of normal rats eating a protein meal. These results suggest that the circadian rhythm in activity of intestinal ornithine decarboxylase is initiated by release of gastrin, or CCK-PZ, or both, and contact of protein with the small intestine after the intake of food containing protein.


Serine protease in mice with hereditary muscular dystrophy. The activities of serine protease, cathepsin B1, ornithine aminotransferase, and aldolase in skeletal muscles of mice with hereditary muscular dystrophy and their normal litter mates were studied. In dystrophic muscle, the specific and total activities of serine protease were much higher than in normal muscle, and the specific activities, but not the total activities, of cathepsin B1 and ornithine aminotransferase were twice those in normal muscle, and several new fragments, which are normally formed by limited proteolysis, were found in dystrophic muscle. When myofibrillar proteins of normal and dystrophic muscles were incubated with highly purified serine protease, their myosin, alpha-actinin and tropomyosin disappeared completely.


Effects of di-(2-ethylhexyl)phthalate administration on carbohydrate and fatty acid metabolism in rat liver. 1. Rats were fed on a diet containing 0, 1, 2, or 4% di-(2-ethylhexyl)phthalate (DEHP), a widely used plasticizer, for four weeks. 2. The level of plasma triglyceride was decreased in both the 1% DEHP and the 2% DEHP groups maintained for 1 to 2 weeks, but that of cholesterol was not changed. Plasma free fatty acid and ketone bodies increased in all DEHP groups. 3. The level of blood glucose was depressed to about 80% in the 2% and the 4% DEHP groups. Liver glycogen decreased markedly. Isotopic incorporations of 14C from [2-14C]pyruvate into blood glucose and liver glycogen were decreased. The changes in the contents of intermediates in the liver indicate that gluconeogenesis is inhibited at the reaction(s) between 3-phosphoglycerate and fructose 1,6-diphosphate. 4. Hepatic fatty acid synthesis increased about 2-fold on DEHP administration. The content of phospholipid in the liver was increased, whereas that of triglyceride was decreased. The rate of phospholipid synthesis was increased, but that of triglyceride was not changed.


Carp connectin: amino acid composition.

Carp connectin: amino acid composition. An elastic protein, connectin, was prepared from carp skeletal muscle by the alkaline method with some modifications; collagen contaminations were exhaustively extracted with 1 N acetic acid and hot phenol treatment was omitted. This connectin preparation contained a considerable amount of tryptophan, but almost completely lacked hydroxyproline.


X-ray analysis of the conformation of chondroitin-4-sulfate calcium salt. The conformation of the chondroitin-4-sulfate calcium salt was investigated by X-ray analysis. The following results were obtained. 1, The repeat length per disaccharide was 0.913 nm: 2, The molecular chain had three-fold screw symmetry: 3, The shape of the unit cell was a trigonal prism with dimensions a=b=1.28 nm, c=2.74 nm, and gamma=120 degrees: 4, The number of disaccharide residues in the unit cell was six. Two molecular chains were packed in the unit cell.


Sodium borohydride as a reducing agent for preparing ninhydrin reagent for amino acid analysis. Sodium borohydride, in place of stannous chloride or titanous chloride, is effective in the preparation of the ninhydrin reagent for automated amino acid analysis. The reagent, coupled with dimethyl sulfoxide as a solvent, a very stable ninhydrin solution which did not form precipitates in the flow lines of the analyzer.


Chemical properties and amino acid composition of beta1-bungarotoxin from the venom of Bungarus multicinctus (Formosan banded krait). beta-Bungarotoxin purified from the venom of Bungarus multicinctus (Formosan banded krait) contained no carbohydrate and behaved as a homogeneous protein on polyacrylamide gel electrophoresis at pH 4.1 and SDS-polyacrylamide gel electrophoresis without 2-mercaptoethanol treatment. Its molecular weight and isoelectric point were estimated to be about 21,000 by gel filtration and about 9.5 by isoelectric focusing, respectively. The toxin treated with the reducing agent was split into two polypeptide chains as revealed by SDS-polyacrylamide gel electrophoresis and their molecular weights were calculated to be about 13,000 and 7,000. The two polypeptide chains (the large one named the A chain and the small one the B chain) were isolated by gel filtration after reduction of disulfide bonds in the toxin followed by alkylation. The A chain contained 120 amino acid residues including 13 half-cystines and the B chain 60 residues including 7 half-cystines. The two chains were supposed to link by disulfide bond(s) in the intact toxin which contained no free sulfhydryl groups. The N-terminal residues of the A and B chains were asparagine and arginine and the C-terminal ones were glutamine and proline, respectively, in accordance with the results of the terminal analyses of the intact toxin.


Purification and characterization of bile acid-CoA:amino acid N-acyltransferase from rat liver. An in vitro study of bile acid-CoA:amino acid N-acyltransferase activity of rat liver was undertaken in order to determine whether separate amino acid-specific enzymes catalyzed the formation of glycine and taurine conjugates of bile acids as postulated by others. Polyacrylamide gel electrophoresis of 200-fold purified enzyme localized the glycine- and taurine-dependent activities to a single band. Both activities were optimal at pH 7.8 and showed similar loss of activity at pH 6.0, pH 9.0, in the presence of 5,5'-dithiobis(2-nitrobenzoic acid), and at temperatures exceeding 50 degrees. With the purified fraction, Km for glycine was 31 mM and Km for taurine was 0.8 mM. Km for several bile acid-CoA substrates was approximately 20 micron and independent of the amino acid acceptor. Only amino acids with terminal alpha- or beta-amino groups were active as acyl acceptors. Acyl donors were limited to bile acid-CoA derivatives. The data support the conclusion that the rat has a single bile acid-CoA:amino acid N-acyltransferase. The substrate kinetics are consistent with previous observations that taurine conjugates predominate in rat bile at normal hepatocellular concentrations of glycine and taurine.


Lysosomal alpha-D-mannosidase of rat liver. Purification and comparison with the golgi and cytosolic alpha-D-mannosidases. Rat liver contains alpha-D-mannosidases in lysosomes, Golgi membranes, and cytosol. The lysosomal enzyme has now been purified approximately 30,000-fold over the crude extract and is free of at least 13 other lysosomal hydrolases. The enzyme has an apparent molecular weight of 335,000 by molecular sieve chromatography and 200,000 by sucrose density centrifugation. It is a glycoprotein, as evidenced by its binding to a concanavalin A affinity column and by a positive periodic acid-Schiff stain. The enzyme has a pH optimum near 4.6. Although it is generally insensitive to a large variety of inorganic salts, chelating agents, and sulfhydryl reagents, prolonged exposure to ethylenediaminetetraacetic acid caused loss of activity, which could be restored by the addition of ZnSO4. Substrate specificity studies were performed on the purified lysosomal alpha-D-mannosidase, as well as on the purified Golgi and cytosolic alpha-D-mannosidases. The three enzymes exhibited only very limited activity on native glycoproteins, but were found to be active on glycopeptides and oligosaccharides, hydrolyzing 1 yields 2 and 1 yields 3 linkages, except that the Golgi enzyme had negligible activity towards the latter linkage. Immunological comparisons by antibody precipitation tests and double-diffusion plates indicated that the three enzymes are not immunologically related. The alpha-D-mannosidase isolated from rat epididymis was found to be immunologically very similar, if not identical, to the lysosomal enzyme isolated from rat liver.


The mechanism of action of ethanolamine ammonia-lyase, an adenosylcobalamin-dependent enzyme. Reaction of the enzyme.cofactor complex with 2-aminoacetaldehyde. Ethanolamine ammonia-lyase (EC 4.3.1.7) catalyzes the adenosylcobalamin-dependent deamination of ethanolamine and 2-aminopropanol. Incubation of the enzyme.cofactor complex with 2-aminoacetaldehyde leads to rapid cleavage of the carbon--cobalt bond accompanied by the destruction of the corrinoid portion of the cofactor. During this reaction the adenosyl portion of the cofactor is oxidized to 4',5'-anhydroadenosine, and the aminoacetaldehyde is converted to acetic acid, which remains associated with the enzyme as a noncovalent complex which survives gel filtration. There is no evidence for the alkylation of the corrin metal by the substrate analog. The enzyme.AdoCbl complex is thus able to eliminate an amino group from a substrate analog without the formation of a new alkyl cobalamin in which the analog is a ligand. These observations do not support the participation of what might be termed "substratylcobalamin" as an intermediate in the ammonia migration occurring in reactions catalyzed by ethanolamine ammonia-lyase.


Expression of functionality of alpha-chymotrypsin. An alternate binding mode in the substrate specificity site. Three-dimensional 2.8 A resolution x-ray crystallographic studies show that toluenesulfonamide and pipsylamide bind isomorphously in the aromatic specificity binding site of alpha-chymotrypsin. However, their orientation differs by about 90 degrees from that usually associated with substrate-like molecules, suggesting a nonproductive binding mode. A secondary binding site is also operative in one molecule of the dimer of the pipsylamide derivative and it is located some 22 A from the active site; however, this site is not operative in the toluenesulfonamide derivative. Binding of toluenesulfonamide and pipsylamide in the specificity site occurs without inducing any significant changes in the native enzyme structure, in contrast to the behavior observed upon tosylation or upon transition state analogue binding of phenylethaneboronic acid. The structural changes accompanying the formation of the latter derivatives are generally asymmetric with respect to the dimeric structure of alpha-chymotrypsin and are generally confined to the binding domain or cylinder 2 of the enzyme (sequence greater than 122). These changes are displayed in a new way via diagonal distance map representation.


Studies on the mechanism of formyltetrahydrofolate synthetase. The Peptococcus aerogenes enzyme. Two conflicting mechanisms have been proposed for formyltetrahydrofolate synthetase (EC 6.3.4.3). Detailed studies with a clostridial enzyme support a sequential mechanism, while a stepwise mechanism with formation of a dissociable intermediate has been proposed for the Peptococcus aerogenes synthetase. However, the data supporting the P. aerogenes mechanism were obtained using synthetase of questionable purity and the results supporting the mechanism could be attributed to contaminating activities. Consequently, uncertainty still exists with regard to the enzyme mechanism. To resolve this uncertainty, the P. aerogenes formyltetrahydrofolate synthetase has been purified to homogeneity and used in experiments to reinvestigate the reaction mechanism. The results of P1:ATP, ADP:ATP, and formate:10-formyltetrahydrofolate exchange experiments as well as a steady state kinetic analysis revealed no difference in the mechanisms of the P. aerogenes or clostridial synthetases. The results are inconsistent with a stepwise mechanism involving a dissociable intermediate and consistent only with a sequential mechanism.


N4-Acetylcytidine. A previously unidentified labile component of the small subunit of eukaryotic ribosomes. The nucleoside content of 18 S rRNA from rat liver is determined under conditions known to prevent the destruction of chemically labile modified nucleosides. Two base-modified nucleosides, not completely identified before, are shown to be N6-methyladenosine and 7-methylguanosine. The results further demonstrate the presence of a hitherto unidentified component of 18 S rRNA whose spectra and chromatographic properties are identical with that of N4-acetylcytidine. In addition, this compound is not detectable in 28 S rRNA nor in 16 S rRNA derived from the small ribosomal subunit of Escherichia coli. However, it appears to be conserved in the small ribosomal subunit of eukaryotes, since it is also present in yeast 17 S rRNA and chicken liver 18 S rRNA.


Amino acid sequence of rabbit skeletal muscle alpha-tropomyosin. The COOH-terminal half (residues 142 to 284). Amino acid sequence analysis of the large cyanogen bromide fragment (residues 142 to 281) derived from the COOH-terminal half of the mixed tropomyosin population of rabbit skeletal muscle has been carried out. The isolation and sequence analysis of peptides derived from chymotryptic digests and from tryptic digests of the maleylated fragment permitted the alignment of the complete sequence except for the assignment of acids or amides at residues 142, 144, and 145. Selected peptides from a Myxobacter 495 alpha-lytic protease digest have confirmed certain overlaps. Based on previously published data the sequence can be extended to residue 284, the COOH-terminal end of the protein. In fourteen positions, amino acid substitutions have been observed. In one of these (residue 199) the sequence evidence indicates a minimum of four different polypeptide chains in the mixed tropomyosin population. The assignment of particular amino acid residues to these positions for the major alpha-component of rabbit skeletal tropomyosin has been based on the relative recoveries of peptides containing different residues in these positions.


The amino acid sequence of rabbit skeletal alpha-tropomyosin. The NH2-terminal half and complete sequence. The amino acid sequence of the large cyanogen bromide fragment (residues 11 to 127) derived from the NH2-terminal half of alpha-tropomyosin has been determined. This was achieved by automatic sequence analysis of the whole fragment as well as manual sequencing of fragments derived from tryptic digestion of the maleylated fragment and thermolytic, Myxobacter 495 alpha-lytic and Staphylococcus aureus protease digestion of the unmodified fragment. Methionine-containing overlap peptides have been isolated from tryptic digests of the maleylated protein as well as from S. aureus protease digests of the unmodified protein. Coupled with previously published information on the small cyanogen bromide fragments and methionine sequences of tropomyosin, these analyses have permitted the completion of the primary structure of the protein. The complete sequence differs by only 1 residue (Gln-24 instead of Glu-24) from that previously reported. Analysis of the sequence by several authors has permitted rational explanations for the stabilization of its coiled-coil structure, for the existence of its two chains in a nonstaggered arrangement, for a head-to-tail overlap of molecular ends of 8 to 9 residues, for the existence of 14 actin-binding sites on each tropomyosin molecule, and a suggestion for the site of binding of troponin-T.


Respiration of hydrogenosomes of Tritrichomonas foetus. II. Effect of CoA on pyruvate oxidation. Hydrogenosomes of the anaerobic flagellate Tritrichomonas foetus can use oxygen as terminal electron acceptor in pyruvate metabolism. In particles broken with the detergent, Triton X-100 or by repeated freezing and thawing, pyruvate-supported respiration is greatly enhanced by coenzyme A. Additions of small amounts of CoA result in a phenomenon resembling respiratory control. ADP and GDP also have a stimulatory effect; however, the rate of respiration in the presence of saturating concentrations of CoA is always much higher than with saturating concentrations of ADP or GDP. The results suggest that the availability of CoA as acetyl acceptor is the primary controlling factor of pyruvate oxidation and that the observed effect of nucleotide diphosphates (Cerkasov, J., Cerkasovová, A., Kulda, J., and Vilhelmová, D. (1978) J. Biol. Chem. 253, 1207-1214) is explained by their participation in reactions that liberate CoA from acyl-CoA complexes in connection with substrate level phosphorylation.


Vectorial production of adenosine by 5'-nucleotidase in the perfused rat heart. Rat hearts were perfused simultaneously with [8-3H] AMP and [8-14C]adenosine. [8-3H] AMP was hydrolzyed by 5'-nucleotidase to produce intra- and extracellular [8-3H] adenosine. Comparison of the specific activities of [3H]- and [14C]adenosine in the heart cells with the specific activities of [3H]- and [14C]adenosine in the effluent perfusate showed that much more [3H]adenosine accumulated in the tissue than would be expected if extracellular adenosine were the immediate precursor of intracellular adenosine. Conversely, perfusion of rat hearts with [8-14C]AMP and [8-3H]adenosine led to a much greater accumulation of intracellular [14C]adenosine than would be expected from an uptake of adenosine from the perfusate. These results are interpreted to be due to hydrolysis of extracellular AMP by 5'-nucleotidase, located in the plasma membrane, and release of the resulting adenosine inside the cell. Measurements of the specific activities of 3H and 14C in ATP, ADP, AMP, and inosine support this interpretation.


A membrane enzyme from Staphylococcus aureus which catalyzes transpeptidase, carboxypeptidase, and penicillinase activities. Staphylococcus aureus H membranes were found to contain four major binding components: Mr = 115,000; Mr = 100,000 doublet; and Mr = 46,000. The low molecular weight protein bound penicillin reversibly and was purified by prebinding membranes with penicillin prior to affinity chromatography. The purified protein catalyzed transpeptidase and carboxypeptidase reactions using di[14C]acetyl-L-lysyl-D-alanyl-D-alanine as the substrate and glycine and hydroxylamine as the acceptors. In addition, the enzyme catalyzed a penicillinase reaction. Kinetic analysis of these reactions revealed similar Vmax values suggesting that, if there is a single active site, the rate-determining steps (i.e. deacetylation) are similar. Rapid denaturation of the enzyme.substrate complex resulted in the detection of covalent penicilloyl- and diacetyl-L-lysyl-D-alanyl.enzyme complexes by sodium dodecyl sulfate-polyacrylamide gel electrophoresis.


Silkmoth chorion proteins. Their diversity, amino acid composition, and the NH-terminal sequence of one component. Silkmoth eggshell (chorion) proteins have been characterized by electrophoresis on sodium dodecyl sulfate and isoelectric focusing polyacrylamide gels; up to 33 and 41 components, respectively, were detected from a single chorion. Some of these components are polymorphic, being absent from chorions of certain animals. A system of nomenclature for all chorion proteins is presented, based on their separation on sodium dodecyl sulfate and isoelectric focusing gels. The chorion is enriched in glycine, alanine, cysteine, and tyrosine and poor in methionine and histidine. The proteins were fractionated into four partially overlapping groups; all four are enriched in the above amino acids, although significant differences exist. Further fractionation by isoelectric focusing of one of the above groups, s/s, yielded seven components, two of which are homogeneous both on sodium dodecyl sulfate and isoelectric focusing gels. The amino acid compositions, molecular weights, and solubility properties of the components share certain features which distinguish s/s as a group from the other three groups. The sequence of the first 67 NH2-terminal residues of a homogeneous protein purified from s/s has been determined. The protein contains a cysteine-rich tail (3 cysteines in the first 18 residues) followed by a 49-residue segment which contains only a single cysteine residue. This latter segment also contains two different tetrapeptide sequences which are each repeated, one twice and the other four times.


Separation of both the Bbeta- and the gamma-polypeptide chains of human fibrinogen into two main types which differ in sialic acid content. The gamma- and Bbeta-polypeptide chains of purified human fibrinogen have each been resolved into two major species: gammaL and gammaR and BbetaL and BbetaR. These molecular variants, separable on CM-cellulose, differ from each other in sialic acid content: approximately 2 residues of sialic acid per molecule of polypeptide chain for the L species to 1 residue of sialic acid per molecule for the R species. The two types of each polypeptide are demonstrable in preparations of fibrinogen from single donors as well as in pooled fibrinogen. The L and R forms of the gamma chains or the Bbeta chains do not differ in their electrophoretic mobility in sodium dodecyl sulfate-polyacrylamide gels, suggesting that they are similar in molecular weight. They are also indistinguishable in polyacrylamide gels in the presence of urea at pH 2.7. Maps of ninhydrin-positive tryptic peptides of the L and R forms of the gamma chain displayed differences within a small group of peptides which have been shown to contain the sialic acid residues present in the gamma-polypeptides. No differences between the peptide maps of BbetaL and BbetaR chains were obvious. A larger ratio of L/R in the gamma and Bbeta chains of dysfibrinogenemia fibrinogen "Zürich II" than in those of normal fibrinogen explains the higher content of sialic acid measured in the native Zürich II fibrinogen molecule.