What administrators look for in their directors of pharmacy. A questionnaire was sent to 160 hospital administrators in Southern California. The questionnaire sought to determine whether directors of pharmacy are good managers, how much time directors spend on administrative tasks, and which managerial skills are most valuable for a director of pharmacy to possess. The results of that questionnaire and a series of recommendations are presented.


The four Cs of food service.

The four Cs of food service. This article focuses on the themes of cost containment, competence, compilance, and commitment in food service administration in the health care setting. Particular emphasis is given to cost containment and various ways to contain costs in the food service operation are presented.


Progress in professional service: nurse leaders queried. Nursing service directors were asked to comment on the progress of key recommendations made by the National Commission for the Study of Nursing in the seven years since the Commission study was published. Their observations and responses are described here, along with conclusions and commentary by the author on the state of affairs in nursing.


Controlling absenteeism can help curb hospitals' costs. Absenteeism involves costs associated with sick pay, overtime pay, decreased employee productivity, and less effective patient care. Hospital management and supervisors must minimize absenteeism and its costs by thoroughly recording and evaluating employees' attendance patterns, counseling and disciplining employees when necessary, and applying policies on attendance with consistency.


System cuts red tape for patients from admission to discharge. One hospital's personalized admitting system is staffed by business representatives who perform a variety of liaison activities to ease patients' nonmedical experience in the hospital. For example, they expedite admissions, discharges, and transfers; make bed assignments; arrange patient transportation; supervise room cleaning on their units; verify patients' insurance coverage; provide financial counseling; and collect payment on bills.


Hospital-based educators take a look at their jobs. Cost containment was high on the list of concerns at the annual meeting and conference of the American Society for Health Manpower Education and Training in New Orleans in June. Speakers exhorted attendees to look at their profession more closely to be sure that they are doing all that they should be doing.


Regulation--Hey! We're in this together. If health care professionals and legislators work cooperatively to set goals and criteria, it may be possible to resolve the confusion about regulations.


Medical students, practitioners hold health fair at high school. This health fair provides a model for introducing health care into a poor, urban, underserved community. Medical students, practitioners, health care institutions, school officials, and local businesses cooperated to provide a program that included physical examinations, educational sessions, entertainment, and information on health careers, especially for young people from "minority groups."


Psychiatric emergency service expands to serve total hospital. A hospital with a comprehensive psychiatric service provides a nearby general hospital with staff and backup for psychiatric consultations, crisis intervention, concurrent psychiatric care of medical-surgical patients, follow-up for psychiatric hospitalization or referrals, and staff education. Personnel include senior psychiatrists and on-call residents, and the program relies heavily on use of psychiatric nursing clinical specialists and RNs with experience in psychiatric nursing.


"Health happening" educates community, enhances hospital's image. A health fair sponsored by St. Mary's Hospital, Lewiston, ME, demonstrated that the public is definitely interested in participating in its own health care. "Health Happening '77" thus proved to be a valuable public service as well as an effective marketing tool and image enhancer for the hospital.


Cardiovascular fitness program--a fringe benefit. St. Marys Hospital Medical Center, Madison, WI, initiated an exercise counseling and activity program for its 1,400 employees. With professional guidance employees are encouraged to become involved in appropriate and enjoyable forms of exercise that will improve and maintain their cardiovascular health and overall well-being. The hospital believes that this is one of the first programs of its kind in a hospital setting.


Medical economics and morality: the conflict of canons. The more the economics of cost containment becomes an ideology, taking on a moral tone, the more it conflicts with the traditional medical morality. Looking at the canons of "good" economics in health care and at the canons of "good" medicine and examining the potential conflicts may point to methods of equilibrating their opposing tendencies.


The last resort: regulation by law.

The last resort: regulation by law. Within the health professions the diminished constraint of conscience and custom upon aggrandizement, acquisitiveness, and exploitation has given rise to the increased role of the government in regulating health care delivery. This situation will continue to exist in direct proportion to the public's convictions that the private sector is incapable of satisfying public expectations. Conversely, when the health care profession, as a whole, recommits itself to a sense of stewardship and accountability to patients and the community, the present system of regulation will pass.


Study shows ESP reduced hospitals' profitability. A study of the Economic Stabilization Program's impact on North and South Carolina hospitals indicates potential effects of more government controls. The study results may be useful in appropriately informing and influencing those pressing for new cost containment legislation.


Study confirms values of ovulation method. In order to determine the effectiveness of the ovulation method in spacing or achieving pregnancy as well as couple satisfaction with the method, the Natural Family Planning  Program of St. Vincent Hospital, Green Bay, WI, studied 453 couples who had participated in the program. The program's director reports that the study results suggest the method provides a safe, effective approach to birth control and assists some women to become pregnant who have not succeeded prior to using this method.


Corporate evangelization: pooling generosity and competence. The conditions of the society in which we live oblige those providing health care to use every means available to bring the Christian message to the people they serve. Only if individuals and religious communities unite in their efforts to provide total care and healing to persons in need can the goal of true evangelization be achieved.


Reorienting staff attitudes toward the dying. After assessing the kind of care it was providing to terminally ill patients and their families, Holy Cross Hospital of Silver Spring (MD) committed itself to a more balanced program of care that included the creation of a thanatology department, implementation of special educational programs for hospital personnel, and exploration of the possibility of establishing a hospice care concept at the hospital.


Functional anatomy of the kidney.

Functional anatomy of the kidney. Initiating a new series of articles on renal function in health and disease, this article describes the ultrastructure of the kidney's basic unit, the nephron, as revealed in recent years by transmission and scanning electron microscopy. With greater appreciation of its morphology and cellular architecture has come new understanding of the functional relationships among the nephron's individual components.


Asthma in adults II: the patient in status asthmaticus. A life-threatening emergency requiring intensive hospital care, status asthmaticus responds to therapy with aminophylline, epinephrine, and steroids. Spirometry and blood gas measurements are used to follow the patient's progress.


Hereditary factors in alcoholism.

Hereditary factors in alcoholism. Being born to an alcoholic parent clearly raises the risk that a man will become an alcoholic, as shown by studies of adopted sons of alcoholic and nonalcoholic biologic parents. Being raised by an alcoholic parent, however, seems not to increase this risk. Alcoholics may possess some sort of biochemical tolerance for alcohol, perhaps based on more efficient detoxification mechanisms than those of moderate drinkers.


Neonatal 'thick blood' syndrome.

Neonatal 'thick blood' syndrome. It has long been known that neonatal hyperviscosity can produce serious central nervous system consequences, including paresis. In recent years, it has been shown that his condition occurs in approximately 5% of newborns and that partial plasma exchange transfusion can lower the hematocrit and may help prevent sequelae. Simple screening of all neonates four hours after birth is recommended so that treatment can be given early.


The Aarskog syndrome.

The Aarskog syndrome. In this report a description is given of the Aarskog syndrome in six males belonging to three different families. Partial expression of the syndrome was confirmed in two of the three examined obligate female heterozygotes, who had short stature, small hands and feet, short neck, and a round face with widow's peak and, in one of them, ptosis of the eyelids.


Are NORs easily translocated to deleted chromosomes? The karyotype of two human meningiomas are reported in which, besides other aberrations, a deleted chromosome 1 and 6 could be observed. In these chromosomes most of the short arm is missing. After silver staining for the detection of NORs, not only the satellite regions of most of the acrocentric chromosomes were stained but also the deleted ends of chromosomes 1 and 6. Besides other explanations for this unusual NOR-staining, the possibility is discussed that NORs of acrocentric chromosomes are easily translocated to deleted chromosomes.


Cytologic observations in 35 individuals with a 5p- karyotype. Chromosome investigation of 35 individuals with a 5p- karyotype and their families revealed the presence of 27 apparently terminal deletions, four interstitial deletions, and four translocations, including two familial cases. Four of the probands with simple deletions and one of the mother were mosaics. Unusual chromosomal heteromorphism, as rendered visible after acridine orange staining, was observed on the short arm of chromosome 14 in two cases and, after heterochromatin staining, on chromosome 19 in one family. Measurement studies, carried out in probands with simple deletions and in two control groups, showed a short-arm loss clustering between 32% and 62% of the normal short-arm length. Using at least two complementary staining methods per proband, we found that the midportion of the 5p15 segment probably must be deleted to develop the typical clinical features of the cri du chat syndrome.


Mitotic cycles in human cell strains with sex chromosomes aneuploidy. A persistence of the embryonic type of mitotic cycle was found in postnatal strains with aneuploidy of sex chromosomes (45,X; 47,XXX; 49,XXXXX; 47,XYY; 49,XXXXY). Life-span and proliferating activity of the strains did not differ from those of diploid postnatal cells.


Triosephosphate isomerase deficiency with hemolytic anemia and severe neuromuscular disease: familial and biochemical studies of a case found in Spain. A 16-month-old girl of Spanish origin with chronic hemolytic anemia and severe neuromuscular disease was found to have markedly reduced triosephosphate isomerase (TPI) activity in her erythrocytes, leukocytes, and plateletes. Both parents and some other family members had moderately reduced erythrocyte TPI activity in accordance with the autosomal recessive mode of inheritance in this enzymopathy. Latex ingestion and latex-stimulated histochemical NBT reduction by the patient's granulocytes were normal. Zymosan-stimulated superoxide radical (O-.2) formation, not previously studied in TPI-deficient granulocytes, was also within normal limits. Starchgel electrophoresis of TPI in both erythrocytes and leukocytes of the proposita and her parents was normal. Molecular studies of deficient TPI showed a normal kinetic pattern with markedly reduced heat instability. Immunologic studies demonstrated no cross reacting material in proposita leukocytes and a normal molecular specific activity. These studies suggest that molecular instability might cause both enzymatic and antigenic degradation of the TPI molecule and, therefore, TPI deficiency in our patient.


Genetic and biosynthetic studies of families carrying hemoglobin J alpha Mexico: association of alpha-thalassemia with HbJ. Hemoglobin J Mexico, an alpha chain mutant, was studied in eight unrelated Algerian families. The quantities of the abnormal hemoglobin in 116 subjects are trimodally distributed: 55% in homozygotes, 31% and 38% in heterozygotes. Both hematological data and the alpha/beta chain biosynthetic ratio are normal in heterozygotes with 31% Hb J and in homozygotes. In contrast, the MCV and MCH as well as the alpha/beta biosynthetic ratio are slightly reduced in heterozygotes with 38% Hb J and in their relatives carrying Hb A. The elevated expression of alphaJ chains in heterozygotes with 38% Hb J may be due to an alpha thalassemia gene trans to the alphaJ locus.


The HLA system and leprosy in Thailand.

The HLA system and leprosy in Thailand. To investigate immunogenetics of leprosy, 205 leprosy patients (26 with tuberculoid, 57 with borderline-tuberculoid, 21 with borderline, 31 with borderline-lepromatous, and 70 with lepromatous leprosy) have been typed for HLA antigens, and compared with 183 healthy controls from the same region (Northern Thailand). There was no significant difference between the overall group of leprosy patients or the three borderline classes and the controls. The two polar forms, tuberculoid and lepromatous leprosy, however, showed significant associations: HLA-A2 is decreased and HLA-Bw17 is increased in tuberculoid leprosy; HLA-B7 is increased in lepromatous leprosy. When both polar forms are compared with each other, HLA-A2 is significantly higher, HLA-Bw40 lower in patients with lepromatous than in those with tuberculoid leprosy. The results are discussed with respect to the different immune responsiveness in the two polar forms of leprosy.


Distribution of G6PD phenotypes in red blood cells of Southern African Negroids: evidence for somatic selection. In a recent population study, we observed a striking deficit of G6PD heterozygotes among Southern African Negroid females. This finding was interpreted tentatively as evidence for a small number of hematopoetic stem cells in man. In a follow-up study we examined peripheral blood and cord blood in 547 mothers and in their newborn offspring. In mothers and sons, the frequencies of the G6PD alleles are apparently quite different. When the allele frequencies determined in sons are used for calculation of the expected phenotype frequencies in mothers and daughters, there is a large deficit of maternal G6PD AB phenotypes, and an equivalent surplus of G6PD homozygotes. However, no relevant heterozygote deficit is observed in newborn daughters. This discrepancy may be explained by the assumption that in peripheral blood of heterozygotes carrying the GdA- allele, G6PD-deficient cells progressively become eliminated during development from birth to adulthood. In other words, the large heterozygote deficit observed in adult females may be due to somatic selection rather than to a small pool of hematopoetic cells at the time of X differentiation.


Interstitial deletion of chromosome 7 detected in three unrelated patients. Chromosome studies were carried out on three patients for the following reasons: (1) growth retardation and mental subnormality in a boy; (2) marked developmental delay in a female infant; (3) routine check on a man whose wife had a stillborn with congenital anomalies. An interstitial deletion at 7q11::7q21 was observed in all three cases.


Familial occurrence of a syndrome with mental retardation, nasal hypoplasia, peripheral dysostosis, and blue eyes in Japanese siblings. Two Japanese siblings, a 2-year-old girl and a 7-month-old boy, had a syndrome of mental retardation, severe nasal hyp9plasia, peripheral dysostosis, and blue eyes. The mother showed nasal hyp9plasia of lesser degree and a mild form of peripheral dysostosis. This disorder bears a striking similarity to acrodysostosis, but in view of certain novel features its relationship to the disease is uncertain. The mode of inheritance could be either dominant with variable expressivity or autosomal recessive.


Assignment of NADH-cytochrome b5 reductase (DIA1 locus) to human chromosome 22. NADH-cytochrome b5 reductase (DIA1, EC. 1.6.2.2) from human fibroblasts and from Chinese hamster cells, both identified by immunologic studies, were clearly distinguished after polyacrylamide gel isoelectro-focusing followed by staining for NADH diaphorase activity. In thirteen independent man-hamster hybrids, the human enzyme DIA1 presented a positive correlation with the human chromosome G22. Eight hybrids were DIA1(+) G22(+) and five hybrids were DIA1(-) G22(-). These data agree with the recent assignment of DIA1 to chromosome G22 by Fisher et al. (1977a). We assume that this newly assigned locus codes for both soluble and microsomal forms of NADH-cytochrome b5 reductase.


10q(q23 leads to qter) duplication: GOTs, HK1, and other gene markers. Gene marker analyses have been carried out in a patient with 10q(q23 leads to qter) duplication. The observed elevation of red cell glutamic oxaloacetic transaminase activity is compatible with earlier somatic cell hybridization studies that mapped the locus to this region. Hexokinase-1 activity in the red cells was normal,which is consistent with its prior assignment to the unaffected part of chromosome 10(10pter leads to q23).


Nucleoli of cultured human lymphocytes. II. Nuceolar fusion and its relation to acrocentric association. Nucleolar fusion and nucleolus formation occurred simultaneously, immediately after mitosis, in cultured human lymphocytes. Evidence is presented that in late telophase and post-telophase, the individual nucleolar organising site includes two components, represented in post-telophase by the nucleolus and its attached process; that fusion can be either directly between the nucleoli or via the processes; and that the latter type of fusion persists into metaphase as acrocentric associations, whilst the former is lost.


Genetic polymorphism of the second component of human complement (C2): presentation of a modified typing technique and data on C2 phenotype distribution, linkage genetics, and haplotype associations a Norwegian family material. Proteins were separated by prolonged isoelectric focusing in polyacrylamide gels, whereupon C2 bands were detected by a specific hemolytic assay. This was performed by treating the gel with iodine to increase C2 activity, and then developing C2 bands with an agarose gel overlay containing sensitized sheep cells and diluted human serum as a complement source deficient in functional C2. The gene frequencies observed in a material of 122 unrelated adults were: C2(1): 0.97 and C2(2): 0.03. C2 linkage relations and C2 haplotype associations have been examined a family material. It is concluded that C2 is very closely linked to HLA loci.


Evidence for a 'silent allele' GLO0 at the glyoxalase I locus. In a three-generation family, the segregation of an apparent silent allele at the GLO I locus in association with the rare HLA haplotype 'AW30-CW4-BW35' was observed in four members. In two cases the assumption of homozygosity at the GLO locus would lead to mother-child exclusions. Phenotypically, the GLO activity in the GLO0 carriers is clearly diminished.


Trisomy-8 mosaicism: report of a case.

Trisomy-8 mosaicism: report of a case. An 8-year-old male with mental retardation, speech difficulties, and minor congenital anomalies is presented. The clinical findings suggest the trisomy-8 syndrome. The karyotype indicates trisomy-8 mosaicism with trisomic as well as normal cell lines in blood lymphocytes.


Patau's syndrome and 13q21q translocation. This paper reports the case of a one-day-old male child presenting the typical features of Patau's syndrome. The cytogenetic study by means of conventional techniques and GTG and QFQ banding techniques showed that the chromosomal pattern of the propositus was 46,XYq+,-21,+t(13q21q) 15ps+,22ps+, and that the nondisjunction that originated the translocation and trisomy had occurred in the mother.


Terminal deletion of the long arm of chromosome 1 in a malformed newborn. Microcephaly and craniofacial dysmorphia, cleft palate, cardiac malformation, and hypospadias are observed in a child with 46,XY,del (1)(q42).


Partial deletion 10q.

Partial deletion 10q. The patient described represents the first reported case of partial deletion 10q. The patient is compared to the partial trisomy 10q syndrome.


Ring chromosome 3 in a retarded boy.

Ring chromosome 3 in a retarded boy. We present a boy with the karyotype 46,XY,r3 and a phenotype with psychomotor and growth retardation, craniofacial anomalies, syndactyly of the toes, and edema of the feet. The karyotypes and phenotypes of both parents are normal.


Balanced and unbalanced pericentric inversion of a chromosome 14. Description of a pericentric inversion of a chromosome 14 with breakpoints at p11 and q23 leading to duplication-deficiency as a result of crossing over.


Investigations on the PGMa1 polymorphism (phosphoglucomutase--EC 2.7.5.1) by isoelectric focusing. The determination of phosphoglucomutase (PGM1) phenotypes was performed by isoelectric focusing on samples from 1678 unrelated individuals from Hessen, Germany. Ten common phenotypes are considered as gene products of four alleles at the PGM1 locus with the following frequencies: PGMa1(1) = 0.6305, PGMa2(1) = 0.1844, PGMa3(1) = 0.1320, and PGMa4(1) = 0.0530. Twenty-two different mating types were observed in 113 families with 202 children. The segregation of the phenotypes in the offspring supports the assumed way of autosomal codominant inheritance. The example of a silent allele (PGM0(1)) as well as a rare variant (PGM7(1)) is reported.


Malate dehydrogenase types in the Asian-Pacific area, and a description of new phenotypes. A survey of more than 21 000 haemolysates from blood samples collected in various parts of south and southeast Asia, Australasia and the Western Pacific and examined in this laboratory has revealed several new alleles controlling variants of sMDH; in addition, further information has been provided on the distribution of sMDH3 in New Guinea. Two of the variant alleles, sMDH3 and sMDH6, achieve polymorphic frequency in various populations. sMDH3 is widely distributed in New Guinea, with highest frequencies in the Eastern Highlands. The pattern of its distribution suggests the mutant arose originally in a Papuan-speaking population. So far, sMDH6 has been detected only in Micronesians from a number of islands in the Carolines. A single example of another new variant, sMDH 5-1, and two examples of a slow variant, sMDH 7-1, were detected in samples from Iran and Singapore, respectively. No examples of mMDH variants were found in a total of 652 placental extracts from Papua New Guinea and Australia.


A variant glucose-6-phosphate dehydrogenase Gd(-) Chiapas associated with moderate enzyme deficiency and occasional hemolytic anemia. Erythrocyte glucose-6-phosphate deficiency is an X-chromosomal-linked hereditary trait often associated with hemolytic anemia. This report defines a new variant designated as Gd(-) Chiapas, which was found in a subject with occasional hemolytic jaundice. The red cell enzyme activity of the subject is about 15% of normal. The variant enzyme is thermolabile in vitro and has faster-than-normal anodal electrophoretic mobility and stronger-than-normal substrate affinity. The patient's hemolytic problem might be correlated with instability of the variant enzyme under physiologic stress.


Transferrin: evidence for two common subtypes of the TfC allele. Evidence is presented for an extended polymorphism of human transferrin (Tf). Three common phenotypes were observed among TfC individuals after isoelectric focusing of sera on polyacrylamide gels. They are explained in terms of two subtypes of the TfC allele, tentatively designated TfC1 and TfC2. The distribution of the phenotypes Tf C1, C2-1, and C2 provides a good fit to the Hardy-Weinberg equation. In our population sample (n = 942) the following frequencies were calculated: TfC1 = 0.8195, TfC2 = 0.1720, TfB2 = 0.0064, TfB1-2 = 0.0016, and TfD1 = 0.0005. Family studies (n = 112) indicate an autosomal codominant way of inheritance. The observed subheterogeneity is detectable in purified transferrin after isofocusing and subsequent immunofixation. The subtypes are still present after treatment of sera with neuraminidase.


Distribution of C3 phenotypes in North India: a pilot study. The commonly occurring phenotypes and some rare variants of C3 were studied in a North Indian population. Based on known gene frequencies, the Indian population seemed more akin to Mongol, African, and Afghan populations than to Caucasians.


DNA, RNA and protein content of tissue during growth and embryogenesis in wild-carrot suspension cultures. A highly selected population of cells (clumps from 63 to 125 micron in diameter), obtained by screening 14-day-old stock suspension cultures of wild carrot (Daucus carota L.), was used to initiate cultures in this study. Time-course changes in DNA, RNA and protein were followed when these cultures were grown in the presence or absence of 2.25 muM 2,4-dichlorophenoxyacetic acid (2,4-D). The data show that growth of these cultures, particularly in the early part of the growth curve, is different from that in most other studies reported on suspension cultures initiated without screening. The gross compositional analysis shows that this difference stems from the very high RNA:DNA and protein: DNA ratios of the cellular material used as the inoculum in this study. The presence of 2,4-D in the medium promoted total RNA and protein levels. Correlations were sought between the appearance of embryos in the absence of exogenous 2,4-D and gross compositional differences developing in cultures grown in the presence and absence of 2,4-D. The handling of cultures during inoculation appeared to have led to a substantial loss of DNA. This had, however, little effect on dry weight or protein content of the tissue.


Expression of fetal antigens in fetal and adult cells during long-term culture. Expression of fetal antigens in early and late passages of tissue culture cells derived from C3H/HeN and C57BL/KaLw mouse fetal cells and from lung tissue of young C57BL/6N mice was investigated by the isotopic antiglobulin technique. The late passage lines of fetal cells had undergone "spontaneous" neoplastic transformation in culture. The antisera were produced by syngeneic immunization with 5000 R x-irradiated tissues from C3H/HeN and C57BL/6N fetuses of 1 to 2 weeks gestation. Fetal antigens were found to be retained even after 5 years in cell lines derived from fetal tissues. In these lines no consistent change in fetal antigen expression could be correlated with neoplastic transformation. In contrast, the early passages of adult cells did not have detectable amounts of fetal antigens. However, fetal antigen(s) was demonstrated in cells of the late passages, and cells of both lines grew as sarcomas when next assayed 55 days later. In addition, fetal antigens were also present in established tumor lines in culture.


Callus formation versus differentiation of cultured barley embryos: hormonal and osmotic interactions. The effect of various plant growth substances as single agents was evaluated in a complex tissue culture system: whole embryo culture of early differentiating barley embryos. Callus formation in unsupplemented medium derives from the mesocotyl and is uniquely characteristic of cultures initiated at this stage of embryonic development. This phenomenon could be prevented or reversed by incorporation of gibberellic acid in the medium resulting in plantlet formation. Indoleacetic acid enhanced callus growth, whereas kinetin did not promote either callus or meristematic development. Callus tissue markedly accumulated starch, effectively lowering the cellular osmolarity, while inducing a corresponding rise in the osmolarity of the culture medium. This osmotic pattern was reversed by gibberellic acid induction of shoot formation. These osmotic-hormonal interactions are interpreted relative to in vivo, in situ normal embryogeny or developmental lesions such as tumors.


Cell cycle analysis and synchronization of the TN-368 insect cell line. A cell cycle analysis of the Trichoplusia ni (TN-368) insect cell line is described. By means of autoradiography and percent labeled metaphase data, the cell cycle parameters were determined to be as follows: S, 4.5 hr; G2, 8.5 hr; M, 0.5 hr; G1, 1.0 hr; the total cell time being 14.5 hr. A synchronization procedure using 50 mM thymidine in a double block procedure was used to provide a method of obtaining a large number of cells in particular cell cycle phases, especially S and G2.


In vitro analysis of proliferating epithelial cell populations from the mouse mammary gland: fibroblast-free growth and serial passage. Normal and neoplastic mouse mammary epithelial cells were cultured in nutrient medium containing D-valine substituted for L-valine. Fibroblast overgrowth was prevented and epithelial cell functions and morphology were retained in cultures maintained in D-valine medium up to 2 months. A nonenzymatic technique was devised to dissociate epithelial cell monolayers. The combined use of this dissociation buffer and D-valine nutrient medium made it possible to passage serially normal and neoplastic mammary epithelial cells. Normal cells were derived from mammary glands of animals stimulated with exogenous hormones for various periods. The period of in vivo hormonal stimulation influenced the ability of normal mammary epithelial cells to attach and proliferate in primary and serially passaged cultures. A greater proportion of cells derived from glands following 2 to 4 weeks of hormonal stimulation were recovered after replating and showed higher labeling indices during serial passage than cells from unstimulated or 5- to 7-week stimulated groups.


Transforming activities of trichloroethylene and proposed industrial alternatives. Three chlorinated hydrocarbons, proposed or already in use as industrial substitutes for the hydrocarbon trichloroethylene, were tested for in vitro transforming potential in a Fischer rat embryo cell system (F1706), which previously has been shown to be sensitive to transformation by chemical carcinogens. Trichloroethylene and the three substitutes (1, 1, 1 trichloroethane, tetrachloroethylene and methylene chloride) all were found to induce transformation, the three substitutes being equal or more efficient transforming agents.


Development of plants from leaf discs of variegated Coleus and its relation to patterns of leaf chlorosis. Leaf discs approximately 8 mm in diameter taken from green and from chlorotic areas of variegated leaves of Coleus were grown in light under sterile conditions in a mineral salt, sucrose, vitamin medium supplemented with auxin and cytokinin. Green shoots, which later formed roots, grew from both green and chlorotic discs in media containing suitable amounts of auxin and cytokinin. None developed in media supplemented with auxin alone or with cytokinin alone. Discs with young plants were transferred to soil. Plants that grew varied widely from those with no chlorosis to those with more chlorosis than the original variety from which the discs were taken. Plants grown from discs taken from green areas of leaves with chlorosis varied in patterns of chlorosis as much as those that grew from discs from chlorotic areas of leaves.


In vitro growth of embryos and callus of coconut palm. A medium for optimal growth of embryos of Jamaican Tall and Green Malayan Dwarf varieties of coconut palm was developed. The Liquid basal Murashige and Skoog medium was supplemented with coconut milk, IAA and 2IP. Activated charcoal improved embryo growth on agar medium. A single callus line was initiated from solid endosperm and subcultured on basal Schenk and Hildebrandt medium supplemented with 2 mg per 1 NAA. Attempts at inducing organogenesis in the callus were unsuccessful. No vascular tissue was present. The callus was aneuploid with the chromosome number = 8 (normal 2n = 32).


Characterization of normal human embryo cells grown to over 100 population doublings. Normal human embryonic cells were subcultured for over 100 population doublings without modification of the basic medium. The cells were evaluated for growth rate, confluent density, chromosome stability, growth in soft agar, ability to hydrolyze casein and tumorigenicity. The cells possessed the characteristics of normal cells. The batch of serum used to supplement the medium was found to be of primary importance in the long-term growth of this cell culture.


A diploid rat liver cell culture. IV. Malignant transformation by aflatoxin B1. Chronic exposure of a cloned rat hepatocyte culture (RL-PR-C) to a subtoxic, sublethal dose of aflatoxin B1 resulted in malignant transformation. Continuous exposure to aflatoxin B1 caused increasing tumorigenic potential as tested by back injection into isogenic animals. Control cultures exhibited spontaneous transformation, although approximately 20 passages beyond the chemically induced event. Neither aflatoxin-treated nor control cultures exhibited cytopathological morphology, formation of cell foci, growth in soft agar, or irregular fibroblast-like growth patterns that could be specifically related to the onset of tumorigenic potential. In general, those parameters commonly used to monitor fibroblast cultures for transformation in vitro were not applicable for assessing the tumorigenic potential of these epithelial cells. Karyotypic analyses revealed no specific chromosomal aberrations associated with aflatoxin treatment; however, chromosomal instability was a property of the tumorigenic cell populations. Injection of both aflatoxin-treated and control cultures at passage 56 resulted in tumors indicative of both carcinoma and sarcoma indicating to us the multipotency of these epithelial cells transformed in vitro.


Influence of 0.5 ppm nitrogen dioxide exposure of mice of macrophage congregation in the lungs. The lungs of 12 mice, half of which were exposed to continuous 0.5 ppm nitrogen dioxide for 3 weeks, were explanted in culture, and the instances of macrophage congregation were quantitated according to numbers of target cells involved, categories of congregation from three to 11 or more, numbers of macrophages participating in each category for the total cultures, and the influence of delaying explantation for 24 and 96 hr. A total of 9042 macrophages and 2140 epithelial and spindle target cells were counted in the outgrowths from 306 explants. The incidence of macrophage congregation (or numbers of target cells) was greater for the cultures from the NO2-exposed animals, both with respect to total incidences between groups (p leads to) and the 0-hr (p less than 0.001) and 24-hr (p less than 0.01) culture subgroups. In addition, the values for T3 to T6 macrophage congregation were individually and consistently greater for the exposed animal group. Postmortem interval stress at 96 hr appeared to result in large colonies, but they were reduced greatly in number. Also the incidence of macrophage congregation fell by 28% as compared to 0-hr and 24-hr subgroups.


Long-term storage of tissue samples for cell culture. The establishment of cultured cell lines from skin biopsies stored at -196 degrees C for periods up to 1 year has been investigated. Attempts to initiate cell cultures from the frozen tissue samples were uniformly successful. There was no alteration in chromosome constitution, morphological appearance, or specific activities of lysosomal enzymes in cells cultured from the stored samples. This process can safeguard against failure of the initial tissue culture and provide an alternate means of storing viable cells when it is impossible or impractical to initiate a cell culture immediately.


Terminal bronchiolar-alveolar (TB-A) units in circumfusion system cultures. Long-term (more than 75 days) recirculating circumfusion system cultures of 15-day fetal mouse lungs gave rise to terminal bronchiolar-alveolar (TB-A) units detached from the respiratory tree. TB-A units established in serum-free microenvironments under reconstructed cellophane membranes became especially flat and optically accessible in fast-flowing 6-chamber systems devoid of a pulse pressure. Time-lapse cinephase movies showed that the alveoli was maintained debris-free by the phagocytosis of alveolar macrophages which entered the alveoli by penetrating the pneumocytic walls. The terminal bronchioles with active ciliary cells communicated with the alveoli and together they formed a flat fluid-filled tubule-like closed system composed of ultrastructurally typical differentiated cellular constituents, viz. types I and II pneumocytes, endocrine-like cells, ciliary cells. Clara cells, and septal cells. TB-A units were observed throughout 150 days of cultivation at which time their phase morphological differentiation remained stable and their functional activity observable as smooth-muscle contractions, ciliation, and alveolar macrophage phagocytosis. A 1000-foot motion-picture film complementing the descriptions of the TB-A units and the alveolar macrophages in the text is deposited with the Motion Picture Film Rental Collection of the Tissue Culture Association, Inc., at the W. Alton Jones Cell Science Center, Lake Placid, New York.


Long-term tissue cultures of human pleural effusions: a cytological follow-up. A cytological follow-up was performed on long-term cultures of 53 metastatic pleural effusions and 34 cases of mesothelial hyperplasia. Cells were grown in Leighton tubes with up to 11 changes of cover slip in the same tube. Cell morphology and arrangement was followed for up to 1 year. Morphological criteria are of no use in recognizing malignant cells with any degree of certainty. As to cell distribution, several types of heaping up are observed both in benign and malignant pleural effusions. In malignant cases, patches of cells in an epithelial-like arrangement are seen heaping up from their substrate. They can be recovered from the used medium by cytocentrifugation. Such behavior may explain the numerous unsuccessful attempts to establish epithelial cell lines, as the clusters of epithelial cells are progressively lost in the supernates.


Hydrocortisone stimulation of human mammary epithelial cells. Rapid proliferation of mammary epithelial cells derived from biopsy specimens of human fibroadenomas was observed when medium was supplemented with ten percent fetal bovine serum and hydrocortisone (5 microgram per ml-1). Hydrocortisone in combination with FBS also led to a 2.5-fold increase in cell cluster attachment and subsequent colony formation. A similar effect was not observed with human serum. In contrast to fibroblast cell systems, insulin did not significantly alter cell growth. The results show that a mitogenic response to glucocorticoids by mammary epithelium may depend on the presence of factors in sera.


Resistance of SJL mice to immunosuppression by antibodies. Immunosuppressive antisera that specifically inhibit induction of tuberculin-type delayed hypersensitivity in mice to the antigens chicken conalbumin and methylated human serum albumin were compared for activity in CF-1, CAF1, and SJL mice. These contrasensitizing antisera were effective in CF-1 and CAF1 mice of various ages. But in SJL mice they were ineffective if the animals were older than 8 weeks and were only moderately effective if the animals were younger. The antibodies of these antisera responsible for specific immunosuppression could be absorbed onto adherent peritoneal exudate cells taken from DAF1 but not SJL mice. They could be eluted from the CAF1 cells at 56 degrees C. These results and other data that are discussed suggest that SJL resistance to immunosuppression by contrasensitizing antibodies is related to a macrophage abnormality. The results also help explain earlier findings that SJL mice are unusually easy to sensitize and unusually resistant to tolerogenesis. This strain of mice should be especially useful for studying the mechanisms of afferent immunosuppression in immunologic enhancement-like forms of tolerance.


Progression of the immune response to solubilized tumor antigens. Local adoptive transfer assays (LATA) were used to analyze and compare the progression of the immune response in C3H/HeJ mice to irradiated tumor cell vaccine and to crude 3M KCl solubilized antigens extracted from a syngeneic methylcholanthrene-induced fibrosarcoma. Sequential LATA performed 2, 6, 9, 12 and 15 days after soluble antigen pretreatment of spleen cell donors revealed a sinusoidal evolution of lymphoid cell activity. An initial brief period of potent tumor facilitation (days 6-9) was followed by a phase of tumor neutralization (days 9-12) which decayed by day 15. On the other hand, spleen cells from donors sensitized with irradiated tumor cells exhibited consistent tumor neutralization which was sustained throughout 15 days. Thus the tumor growth facilitation observed only in CSA-treated donors may represent a qualitative difference in the immune state induced by soluble, as opposed to cellular, forms of tumor antigen.


The A, M, N, and P1 blood group substances in butanol extracts of human erythrocytes. Aqueous phase and butanol phase extracts of group A1, O, M, N, P1 and P2 human erythrocytes perpared by extraction of red cell stromata with n-butanol were examined for the presence of blood group substances in inhibition of agglutination experiments using antisera and test cells of human origin. Group A substance was recovered in both aqueous phase and butanol phase extracts, M and N substance in aqueous phase extracts only, and P1 substance only in butanol phase extracts. Use of high concentrations of extract resulted in detection of P1 activity in aqueous phase extracts and of N specific inhibition in the aqueous phase extracts of group M erythrocytes. Distilled water appeared to be superior to digitonin and hypotonic phosphate buffer solutions for preparation of stromata for the recovery of group A and M substances in the butanol extracts but not for group N and P1 substances.


Colonization of teeth in humans by Streptococcus mutans as related to its concentration in saliva and host age. The relationship of the salivary concentration of Streptococcus mutans and host age to the colonization of this organism on erupting teeth was studied in humans. Plaques were obtained from fissures and buccal surfaces of erupting permanent first and second molars of children 6 to 7 and 11 to 14 years old, respectively. In subjects of both age groups with salivary S. mutans concentrations below 5 x 10(2) colony-forming units per ml, the organism was detected on only a few of the tooth surfaces; at concentrations of 5 x 10(2) to 4.9 x 10(4) colony-forming units per ml more than half of the surfaces and at concentrations of 5 x 10(4) colony-forming units per ml or higher most of the surfaces were colonized by S. mutans. The frequency of detection and concentration of S. mutans in plaque as well as its concentration in saliva were higher in the case of the older children. However, when younger and older children with similar salivary S. mutans concentrations were compared, S. mutans was more frequently isolated from plaque from older children only in the case of children with below 5 x 10(2) colony-forming units per ml of saliva. Eleven of the 64 children studied had low or undetectable S. mutans levels in plaque and saliva. The salivary S. mutans levels of the parents of these children were lower than those of parents of a group of children with normal oral S. mutans levels.


Experimental pathogenicity and mortality in ligated ileal loop studies of the newly reported halophilic lactose-positive Vibrio sp. Laboratory animals were challenged subcutaneously, intraperitoneally, and intravenously with the halophilic lactose-positive Vibrio. Intraperitoneal inocula of 10(8) organisms proved to be rapidly lethal in mice, rats, and hamsters. The 50% lethal doese in ICR strain mice was estimatd to be 8 X 10(5) live cells, injected intraperitoneally or subcutaneously. Subtaneous inocula in mice resulted in severe local infections, characterized by gross edema, and for those animals surviving longer than 48 h, tissue necrosis. Intravenous, intraperitoneal, and subcutaneous injections of 10(8) cells in mice resulted in death within 3 to 6 h. These animals rapidly developed Vibrio bacteremia after injections. V. parahaemolyticus, studied for comparative purposes, produced no morbidity or mortality when injected subcutaneously. Injections of live lactose-positive Vibrio into ligated ileal loops in rats and rabbits consistently proved to be lethal with a high-density bacteremia resulting.


In vitro and in vivo photosensitized inactivation of dermatophyte fungi by heterotricyclic dyes. The ability of three heterotricyclic dyes to photosensitize dermatophyte fungi was studied with Trichophyton mentagrophytes and Microsporum gypseum. In vitro studies showed that methylene blue, neutral red, and proflavine were capable of killing these fungi when used in conjunction with broad-spectrum light. Proflavine, however, killed both fungi most rapidly and was used for further studies. Fungal killing by proflavine plus light was dependent on dye concentration, pH, light wavelength, and light intensity. Based on the in vitro studies, a treatment regimen was developed for in vivo use on experimentally infected animals. When treatment of guinea pigs inoculated with T. mentagrophytes was begun during fungal invasion, lesion formation at inoculated sites was either prevented or substantially reduced. When treatment was begun after lesion formation, however, light-plus-dyed treated sites showed only slightly faster curing than untreated sites.


Molar growth yields and enterotoxin B production of Staphylococcus aureus S-6 with amino acids as energy sources. Staphylococcus aureus S-6 was able to utilize glutamate, proline, histidine, aspartate, alanine, threonine, serine, or glycine as a major energy source, when these were added singly at a concentration of 2.5 g/liter to a base medium containing salts, vitamins, and 30 mg of each of 18 amino acids per liter. Molar growth yields (Y) for each of the eight amino acids were determined in aerobic batch cultures. Except with aspartate and serine, all molar growth yields decreased with increasing concentration of the energy source, whether or not it was present in excess. The average molar growth yields for both aspartate and serine were Y = 35 g/mol. The highest Y values obtained with the other amino acids were 79, 78, 56, 46, 58, and 21 g/mol for glutamate, proline, histidine, alanine, threonine, and glycine, respectively. Histidine was not utilized as an energy source at concentrations of less than 2 mM. Enterotoxin yields (mg of toxin per g [dry wt] of cells) higher than in the basal medium were obtained when proline, histidine, alanine, serine, and glycine were the major energy sources. When proline and alanine were the major energy sources, enterotoxin yields were not affected by the concentration, whereas toxin yields were reduced at high concentrations of the other six amino acids. The highest cell yields and Y values were obtained with glutamate, but its utilization as an energy source markedly reduced enterotoxin yields.


Effect of minerals on staphylococcal enterotoxin B production. Raising the magnesium level from 0.4 to 1.5 mM in a medium containing only amino acids (2.08%), salts, and vitamins increased enterotoxin B production by Staphylococcus aureus S-6 by about 80%. The level of phosphates in the medium was lowered to one-tenth (to 2.87 mM) the original amount without adversely affecting growth and enterotoxin production. The optimum level of potassium was between 15 and 45 mM. Ammonium salts were not essential in the amino acid medium. Sufficient quantities of trace metals were already present. Revising the salt mixture according to the information obtained resulted in a doubling of the enterotoxin B production.


Ultrastructural studies on dengue virus infection of human lymphoblasts. Ultrastructural studies of dengue-2 virus-infected lymphoblastoid Raji cells showed that the virus induced an increase in the size of the rough endoplasmic reticula (RER) and that the replication of the virus was confined to the cisternae of these RER. The proliferating RER formed cytoplasmic inclusions that could be seen by light microscopy. This observation could be used as evidence of a cytopathogenic effect of dengue virus on infected Rajii cells in routine cultures. Accumulation of virions in the infected cells was minimal in comparison with other cell systems, however. Sporadic clusters of mature virions were often seen on the plasma membrane. These extracellular virions were distributed adjacent to the virus-bearing RER and were presumably released virions. Vertical transmission of the virus was evident in mitotic lymphoblasts. The replication pattern of dengue virus in lymphoblastoid cells suggests that efforts should be made to determine whether blast-transformed lymphocytes, numerous in secondary dengue infections, support dengue virus replication in vivo.


Substances that interfere with action of viridin B, a Streptococcus mitis bacteriocin. Several body fluids and various substances were examined for their effects on the action of viridin B, a Streptococcus mitis bacteriocin. Heme has been shown previously to interfere with viridin B action. In the present study, peroxidase and catalase, but not superoxide dismutase, interfered with the bacteriocin. Sulfhydryl-containing compounds had no effect. Trypsin, protease, and amylase exhibited interfering capacities. Of various body fluids tested, serum, saliva, and leukocyte lysate interfered with viridin B. The activity in leukocyte lysate was probably due to myeloperoxidase. Salivary activity was likely due to both peroxidase and amylase. Interfering activity in serum was neither complement dependent nor antibody mediated and was probably due to a combination of enzymatic activities. Temporal studies with these agents suggested more than one mechanism of interference with viridin B.


Induction of a pH-stable interferon in sheep lymphocytes by Mycoplasmatales virus MVL2. The data presented in this report show that the enveloped Mycoplasmatales virus MVL2 is capable of interferon induction in sheep peripheral blood lymphocytes. The phenomenon was dose dependent, requiring approximately 10(8) plaque-forming units of virus per 2 X 10(6) lymphocytes. The interferon was stable to pH 2.0 treatment, was produced in moderately high levels (greater than 1,000 units), and met many of the criteria for classification as a type I interferon. Heat-inactivated MLV2 lost its ability to induce interferon, whereas ultraviolet-inactivated virus retained its capacity to induce levels comparable to untreated virus. Whereas the MVL2 used in these studies was contaminated with several host cell proteins as determined by sodium dodecyl sulfate-containing polyacrylamide gel electrophoresis, the contaminants probably did not play a role in the induction because isolated cell membranes or soluble cell contents from Achoplasma laidlawii are inactive as inducers. Also presented in this report is a preliminary description of the structural polypeptides of MVL2.


Lipid metabolism of Borrelia hermsi.

Lipid metabolism of Borrelia hermsi. The synthesis of complex lipids by Borrelia hermsi while growing in Kelly medium was investigated by labeling cultures with D-[14C]glucose or [14C]palmitic acid. Labeled glucose was incorporated into phosphatidyl choline, phosphatidyl glycerol, monogalactosyl diglyceride, cholesterol glucoside, and acylated cholesteryl glucoside. The fatty acid composition reflected that of the medium suggesting that this spirochete directly incorporates acyl chains unaltered from its external milieu. Furthermore, the distribution of labeled carbon between acyl groups (lipid-soluble) and water-soluble moieties indicates that there is no metabolic exchange between these two pools. The relationship of B. hermsi to other spirochetes is discussed in terms of lipid metabolism as a generic characteristic.


Age-related susceptibility to Pseudomonas aeruginosa ocular infections in mice. The susceptibility of newborn and infant mice to eye infection by Pseudomonas aeruginosa was studied in 5-, 10-, 15- to 16-, and 21-day-old mice. In the first of three age-related susceptibility experiments, inoculation of P. aeruginosa under the unopened eyelids of infant (5- and 10-day-old) mice in the absence of prior corneal wounding resulted in acute infection and rapid death of many of the animals. However, endophthalmitis was observed in about 30% of bacteremic animals that survived to age 14 to 15 days. In the second experiment, 15- to 16-day-old mice whose eyes were open received P. aeruginosa topically onto either wounded or unwounded corneas. At least 50% of the mice that received both corneal wounding and the bacteria exhibited keratitis, endophthalmitis, and subsequent phthisis bulbi. None of the infected mice died of bacteremia. In addition, mice infected in the absence of corneal wounding did not exhibit any eye damage. In the third experiment, the wounded-cornea responses of 21-day-old mice to P. aeruginosa were more variable. Thirty seven percent of the mice exhibited an intermediate response of decreased eye size and cataracts which was not observed in 15- to 16-day-old mice, 32% recovered spontaneously, and 29% exhibited complete shrinkage of the infected eyes. The variability of the latter responses may reflect a transitional maturation period of natural immunity to the organism in some of the animals, since all 4- to 6-week-old adult mice respond routinely to ocular wounding and similar infections with the organism by undergoing a spontaneous resolvable keratitis (3 to 4 weeks).


Chromosomal locus for staphylococcal enterotoxin B. The genetic locus of staphylococcal enterotoxin B (SEB) was investigated in the Staphylococcus aureus food-poisoning isolates, strains S6 and 277. Direct neutral sucrose gradient centrifugation analysis of sodium dodecyl sulfate-sodium chloride-mediated cleared lysates demonstrated that strain S6 contained a single 37S plasmid. Transductional analysis revealed that the 37S plasmid in S6 encoded for cadmium resistance (Cad) but not SEB. Additionally, elimination of cadmium resistance in S6 provided a plasmid-negative derivative that produced SEB at the same level as the parent. Examination of strain 277 showed two plasmids, a 37S species encoding for penicillin resistance (Penr) and a 21S species containing the gene(s) responsible for tetracycline resistance (Tetr). Elimination of the 37S, penr plasmid in 277 had no effect on SEB production, whereas introduction of the 21S tetr plasmid via transformation into strain 8325 (SEB--) did not confer enterotoxigenesis upon the transformants. The data obtained in this investigation suggest that the SEB gene(s) in these food-poisoning isolates of S. aureus is chromosomal.


Evidence for a disseminated plasmid in Streptococcus mutans. Based on a survey of 86 isolates, approximately 5% of all naturally occurring strains of Streptococcus mutans contains a 3.6 X 10(6)-dalton (3.6-megadalton) multicopy plasmid of unknown function. The amount of plasmid deoxyribonucleic acid per chromosome varies from 2 to 6% depending on the host strain. About 13% of the total covalently closed circular deoxyribonucleic acid in each of the four plasmid-containing strains consists of dimeric molecules, with interlocked circular forms predominating. Site-specific restriction endonucleases have been identified that cleave this 3.6-megadalton plasmid at single and at multiple sites. Each of the four plasmids is cleaved once by the HindIII and BamHI restriction enzymes. The HpaI, TaqI, and HhaI enzymes generate two, five, and six components, respectively, and the digestion products of each of the four plasmids are identical. Because the four plasmid-containing S. mutans strains are physiologically unique with respect to one another, we conclude this plasmid to be a disseminated extrachromosomal element in S. mutans.


Isolation of a bacteriophage for actinomyces viscosus. A lytic phage which produces clear plaques on a human isolate of Actinomyces viscosus was isolated from a sample of raw domestic sewage.


Rapid activation of the interferon system in vivo. Experiments were carried out to study the kinetics of local interferon production in the subcutaneous tissues of rats stimulated with Newcastle disease virus. Specifically, the interferon produced and released in the extracellular fluids was collected at various intervals of time in micropore chambers implanted into the subcutaneous tissue of rats. Interferon was detected at moderate titers 1 h after induction, and it was present at high titer at 2 h. The interferon levels remained remarkably high in the samples collected after 3, 5, and 24 h, and in some rats it was still detectable after 48 and 72 h. Since control experiments showed that it requires 2 to 3 h for interferon to penetrate the chambers, it may be concluded that high concentrations of interferon are present in the extracellular fluid within 1 h of induction. The evaluation of the kinetics of production and of the concentrations attained in the extracellular fluid suggests that in a solid tissue a cell infected by a potent interferon inducer may produce interferon early enough and in sufficient quantity to protect neighboring cells before the production of progeny virions.


Innate resistance of mice to experimental infection with Naegleria fowleri. The mouse system provides an excellent model for studying host resistance to Naegleria fowleri, the agent of primary amoebic meningoencephalitis. Innate resistance to infection with N. fowleri was examined with respect to infecting dose and the age, sex, and strain of mice. Intravenous inoculation with 10(7) amoebae per mouse produced 100% mortality in 9 days, whereas inoculation with fewer amoebae reduced the cumulative mortality. Male and female DUB/ICR mice of varying ages were inoculated intravenously with 2.5 X 10(5) N. fowleri per g of body weight. The youngest mice died first, with 100% mortality for both males and females, and mortality decreased with increasing age. Female mice were significantly more resistant to infection than males. Five strains of mice weighing approximately 20 g were inoculated intravenously with weight-adjusted doses; mortality ranged from 10% in C57BL/6 mice to 95% in A/HeCr mice.


Cross-protection of mice provided by active and passive immunization against experimental infections with virulent Proteus rettgeri and Providencia bacteria. Immunization with Providencia and Proteus rettgeri Formalin-treated bacterial suspensions produced high levels of protection in mice against homologous and heterologous challenge. Mice were also cross-protected, but less effectively, by passive administration of rabbit type-specific antisera. The protective activity appeared to be due to an antigen common to strains of different O-serotypes. It was not detectable in agglutination reactions, and preliminary results indicate that it is thermostable, not being inactivated in its antibody binding capacity at 121 degrees C for 1 h.


Inverse relationship between the susceptibility of lipopolysaccharide (lipid A)-pretreated mice to the hypothermic and lethal effect of lipopolysaccharide. Mice pretreated (day 0) by a single injection of lipopolysaccharide (LPS) responded with hypothermic tolerance to (LPS) challenge on day 1 and with hypothermic hyperreactivity to LPS challenge on day 4. Reciprocally, mice pretreated similarly but with a higher challenge dose were hyperreactive with respect to LPS lethality on day 1, but highly tolerant to lethality when challenged on day 4. Hyperreactivity to LPS lethality (day 1) was evident from an accelerated onset of death as well as from a reduced 50% lethal dose in pretreated mice, the level of hyperreactivity being more pronounced with higher LPS pretreatment doses. Lethal hyperreactivity, however, was only seen after challenge with a 50% lethal dose of soluble LPS. In contrast, protection to lethality occurred after challenge with a 50% lethal dose of insoluble LPS (day 1). Tolerance to LPS lethality in mice was observed on day 4 after pretreatment with one (day 0) or four daily injections of LPS. Since reciprocal hyperreactivity (day 1) and cross-tolerance to lethality (day 4) could be achieved by treatment with Salmonella smooth- or rough-form LPS as well as with free lipid A, it was concluded that lipid A represents the active principle of LPS in inducing both hyperreactivity and tolerance to the lethal effect of LPS.


Activation of complement by cell surface components of Staphylococcus aureus. The abilities of intact Staphylococcus aureus H, crude cell walls (CCW), purified cell walls (PCW, peptidoglycan [PG] and covalently linked teichoic acid), peptidoglycan, and cell membranes (CM) to activate the complement system in normal human serum, C2-deficient serum, and immunoglobulin-deficient serum were compared. On a weight basis, PCW was the most active fraction; intact organisms and CCW were about equally effective; and PG was least active in causing complement consumption in normal serum. CM also activated complement but did not give a clear dose-response relationship in the concentrations used. Kinetic studies revealed that C3-C9 consumption occurred at a significantly slower rate in C2-deficient serum, indicating that intact organisms, PCW, and PG may activate the complement system via the classical and alternative pathways in normal serum. C3-C9 consumption was also slower in immunoglobulin-deficient serum than in normal serum, implying that immunoglobulins play a role in attaining maximum rates of complement activation. In all sera studied, PG was less active in complement activation than PCW. These results indicate that a number of cell surface components of S. aureus can play a role in complement activation by this organism and that the presence of teichoic acid has a significant enhancing effect in this regard.


Immune reactivity of Visna virus-inoculated mice. Visna virus inoculation of mice before immunization with unrelated antigens caused suppression of humoral and cell-mediated immune responses. The immunosuppressive effect was dependent on the virus dose and time of administration. The number of "background" antibody-producing spleen cells increased slightly after inoculation of Visna virus. Heat treatment of Visna virus abolished its immunosuppressive effect. Replication of Visna virus in mice seemed to be a prerequisite for its immunomodulating effect.


Delayed-type hypersensitivity in mice immunized with Trypanosoma rhodesiense antigens. When mice were immunized intravenously, subcutaneously, or by the footpad route with formaldehyde-killed Trypanosoma rhodesiense, delayed-type hypersensitivity was elicited by the use of frozen-thawed trypanosomal antigen. The delayed footpad swelling technique was used to measure delayed hypersensitivity. Hypersensitivity induction was dose dependent (greater than or equal to 10(6) formaldehyde-treated T. rhodesiense) and was affected by the route of immunization. The footpad route induced higher levels of hypersensitivity than other routes of immunization. Mice immunized with a single dose of formaldehyde-treated antigen and challenged with live T. rhodesiense did not survive. Yet, mice immunized subcutaneously with formaldehyde-treated antigen and then injected with frozen-thawed antigen and challenged 28 days after immunization survived. The results suggest that T-cell activation, manifested by delayed hypersensitivity responses, was a necessary component in the protective response, perhaps functioning in a helper cell capacity.


Purification and characterization of an immunologically distinct delta-hemolysin from a canine strain of Staphylococcus aureus. Culture supernatants of 26 strains of Staphylococcus aureus possessing delta-hemolytic activity have been tested by immunodiffusion against a serum raised against purified delta-hemolysin from S. aureus CN 4108 (Newman D2). Supernatants from 14 strains of canine origin gave a reaction of partial identity with delta-hemolysin from CN 4108, whereas supernatants from all other strains had full identity. Delta hemolysin from one of these canine strains, CN 7450, was purified by ammonium sulfate fractionation and precipitation at pH 4.5. The physical, chemical, and biological properties of this toxin were compared with those of delta-hemolysin from CN 4108. Differences in molecular weight (as judged by Sepharose 6B chromatography), isoelectric point, and amino acid composition were found. Both toxins caused dermonecrosis in rabbits, lysed erythrocytes from several different species, and were inhibited by normal sera and phospholipids. Unlike delta-hemolysin from CN 4108, the hemolytic activity of delta-hemolysin from CN 7450 was found to be dependent on the incubation temperature over the range of 25 to 37 degrees C. Immunodiffusion results obtained with antisera raised against purified delta-hemolysin from CN 7450 indicated that delta-hemolysins from the canine strains were probably identical and confirmed that these differ immunologically from delta-hemolysin from CN 4108.


Cytopathogenicity of Mycoplasma hyopneumoniae in porcine tracheal ring and lung explant organ cultures alone and in combination with monolayer cultures of fetal lung fibroblasts. Porcine tracheal rings and lung explants alone and in combination with monolayer cultures of porcine lung fibroblasts (PLF) were separately inoculated with virulent strains of Mycoplasma hyopneumoniae and incubated at various times. The preparations were observed by bright-field, phase-contrast, and scanning electron microscopy. In PLF cultures, the strains at initial concentrations of 10(1.3) colony-forming units/ml increased within 3 days to 10(6) colony-forming units/ml, showed progressive clustering on the cells, and caused some sloughing. Introduction of a tracheal ring or lung explant into these mycoplasma-infected PLF cultures caused the explant to lose its epithelial ciliary motility. Eventually parts or whole cells of the respective ciliated epithelium were lost. Without infected PLF monolayers, the explants inoculated with M. hyopneumoniae were less susceptible to infection. When uninfected explants were incubated for 18 days or kept in stock for 2 months, they did not show the above changes. With 5 h postinoculation, M. hyopneumoniae cultures became intimately associated with the PLF culture, but when epithelial cell sloughing occurred, the mycoplasmal cells became dependent on the introduction of a fresh PLF monolayer or a tracheal or lung explant for survival.


Suramin effects on macrophage phagolysosome formation and antimicrobial activity. The effects of suramin on phagolysosome formation and antimicrobial activity of mouse peritoneal macrophages cultivated in vitro have been studied. Prolonged in vitro pretreatment of macrophages with high concentrations of suramin caused macrophages to form large fragile phagolysosomes in which the concentrations of the various lysosomal enzymes were inferred to be diminished. In addition, suramin-treated macrophages demonstrated enhanced exocytosis of acid phosphatase during phagocytosis of polyvinyl toluene spherules. However, suramin was found not to inhibit formation of phagolysosomes in macrophages that had ingested Listeria monocytogenes when those cells were examined by the electron microscope. Suramin pretreatment did not alter the ingestion or intracellular killing of Staphylococcus aureus or of a strain of L. monocytogenes that was essentially avirulent for mice, but did protect macrophages from destruction by virulent L. monocytogenes ingested in vitro, an effect that appeared to have been mediated through enhancement of the bacteriostatic potential of the macrophages. However, at a single dosage level, the drug did not alter the mortality of mice challenged with virulent L. monocytogenes.


Characterization of Thermoactinomyces sacchari antigens. Three different extracts of Thermoactinomyces sacchari were analyzed for their antigenicity and physical-chemical properties. Rabbit antiserum to each preparation, tested by immunodiffusion in gel, demonstrated that the most potent immunogen was that prepared by the double-dialysis method. This extract also contained the greatest number of precipitating antigens as detected by gel filtration. All extracts, analyzed by column chromatography on Sephadex G-75, were heterogeneous in that they contained large- and small-molecular-weight fractions. Precipitins in each extract were detected in column eluates of relatively low ultraviolet absorption and were of a similar molecular size range. Chemical analysis of purified antigens demonstrated protein and carbohydrate. The culture supernatant contained the greatest amount of carbohydrate (66%), and the soluble extract of bacterial cells contained the greatest amount of protein (68%). Several antigens were partially sensitive to the proteolytic enzymes Pronase and trypsin, but none was sensitive to lysozyme. These results demonstrate that there are multiple-antigen systems in T. sacchari and that, of the samples analyzed, the double-dialysis method of antigen preparation yields the most potent antigens.


Clindamycin-induced enterocolitis in hamsters as a model of pseudomembranous colitis in patients. Stools from a patient with antibiotic-associated colitis and cecal contents from a hamster with clindamycin-induced enterocolitis were compared in a cytotoxicity assay to determine common properties. Both specimens produced actinomorphic changes in human amnion cells at 10(-7) dilutions. The toxin was acid labile, heat labile, nonether extractable, non-dialyzable, and produced maximum activity at 60% with ammonium sulfate precipitation. Cytotoxicity was neutralized with clostridial antitoxin but not with equine serum. Clostridium difficile was recovered in high concentrations in specimens from both the hamster and patients. The supernatants of these C. difficile strains produced cytoxic effects which were also neutralized by clostridial antitoxins. These results indicate that clindamycin-induced enterocolitis in hamsters is a model of human disease and implicate toxin-producing clostridia as responsible agents.


Antigenic analysis of coccidioidin and spherulin determined by two-dimensional immunoelectrophoresis. Immunological tests are valuable aids for diagnosis of mycotic infections and, in some cases, as objective guides for clinical management and prognosis. The usefulness of these procedures is limited to the extents that crude antigen preparations are employed, that these are difficult to standardize uniformly, and that they contain antigens common to several species of pathogenic fungi. Analysis by two-dimensional immunoelectrophoresis methods of the two crude preparations used for coccidioidomycosis demonstrated that coccidioidin contained at least 26 antigens, with 10 of these found also in spherulin. In addition, spherulin contained two antigens not demonstrated in coccidioidin. No single test detected all antigens present, and multiple procedures were required to display the complete array of antigens. A reference system was established for coccidioidin and precipitated immunoglobulins from a burro hyperimmunized with coccidioidin. Evaluation of the reference system demonstrated that it was highly reproducible with respect to the reagents used, to repeated tests by the same person, and to comparative tests by two individuals using the same reagents. Applications of this reference system for standardization of reagents, for detecting common antigens, for monitoring successive steps during fractionation of crude preparations, and for fingerprinting strains for ecological and epidemiological studies are presented.


Purification of cholera toxin and its subunits: new methods of preparation and the use of hypertoxinogenic mutants. Cholera toxin was obtained in pure form by fractionation on two phosphocellulose columns successively. Cholera toxin and choleragenoid were quantitatively and selectively adsorbed to the first column in 10 mM phosphate buffer, pH 7.0, and were subsequently eluted with buffer of high ionic strength. The toxin was then separated from choleragenoid on the second column by chromatography at pH 8.3. The toxin obtained was highly active and pure as judged by electrophoresis, isoelectric focusing, and various immunological and chemical tests. Pure choleragenoid was by-product of the procedure. The A1 chain of the toxin was obtained in pure form by treating phosphocellulose-bound toxin with urea and a reducing agent. The anionic A1 peptide was thereby released, leaving a complex of the B and A2 chains (A25B) bound to the resin. The latter was then eluted and further purified to obtain nontoxic antigen. The overall yields of cholera toxin and choleragenoid were increased two- to threefold by the use of hypertoxinogenic mutants of Vibrio cholerae.


Invasive aspergillosis: antiserum for circulating antigen produced after immunization with serum from infected rabbits. We describe a method for producing an antibody capable of detecting an antigen of Aspergillus fumigatus in serum and urine from infected rabbits and humans.


Effect of sucrose in culture media on the location of glucosyltransferase of Streptococcus mutans and cell adherence to glass surfaces. Streptococcus mutans strain B13 (serotype D) almost exclusively produced free glucosyltransferase (GTase) in the culture supernatant when grown in sucrose-free TTY broth medium, which was composed of Trypticase (Baltimore Biological Laboratory [BBL] Cockeysville, Md.), tryptose (Difco Laboratories, Detroit, Mich.), yeast extract (BBL), salts, and 1% glucose. Organisms grown in sucrose-free TTY broth retained very weak cell-associated GTase activity and did not adhere significantly to glass surfaces in the presence of exogenous sucrose. If sucrose was added to TTY broth, however, GTase was found on the cell surface where cell-bound, water-insoluble glucans were synthesized. Most commercially available products of Todd-Hewitt broth were found to contain trace amounts of sucrose, as did Trypticase soy broth (BBL), whereas brain heart infusion broth (Difco and BBL) was found to be essentially free of sucrose. Almost all detectable GTase activity was cell associated when S. mutans B13 was grown in Todd-Hewitt or trypticase soy broth. Heat-treated B13 cells grown in Todd-Hewitt broth and cell-free, water-insoluble glucans bound free GTase and produced marked adherence in the presence of sucrose. Experiments strongly suggest that the binding sites for free GTase are the surface glucans, and cell-associated and extracellular GTases are most likely alternate states of the same enzyme protein.


Contrast of Glycogenesis and protein synthesis in monkey kidney cells and HeLa cells infected with Chlamydia trachomatis lymphogranuloma venereum. Glycogen metabolism of monkey kidney (LLC-MK-2) cells and HeLa 229 cells infected with a Chlamydia trachomatis lymphogranuloma venereum 440 L (LGV) was studied. The growth cycle of LGV in both host cells was similar; however, a greater number of infectious organism developed intracellularly and were released into the medium during LGV infection of HeLa 229 cells than MK-2 cells. A rapid infection accompanied by a high rate of glycogen synthesis and a short period of accumulation was found in GeLa 229 cells infected with LGV. LGV infected MK-2 cells started to accumulate glycogen about the same time as HeLa 229 cells; however, the rate of glycogen synthesis was lower and the period of accumulation was longer. The LGV agent grew in cycloheximide-treated cells in the absence of host cell protein synthesis. Protein synthesis associated with LGV throughout the developmental cycle was similar in both cell types and could be abolished by chloramphenicol. The continued synthesis of glycogen in the presence of cycloheximide suggested that the synthesis of glycogen was directed by the organism in both MK-2 cells and HeLa 229 cells.


Lymphocyte cytotoxicity to influenza virus-infected cells: response to vaccination and virus infection. Peripheral blood leukocytes obtained from volunteers at various times following influenza vaccine or live influenza virus infection were assayed for cytotoxicity against influenza virus-infected cells. Cytotoxicity was highest on days 3 and 7 following vaccination with commercial A/Port Chalmers/1/73 inactivated influenza virus vaccine. Maximal cytotoxicity was found 9 days after infection induced by intranasal inoculation of a strain of A/Scotland/840/74 influenza virus. Individuals naturally infected with A/Victoria/3/75 were also found to have elevated cytotoxicity approximately 1 week after onset of illness. Cytotoxicity levels decreased toward base line approximately 30 days after the virus exposure. The effector mechanism(s) responsible for the early, transient elevation in specific immune release to influenza virus-infected cells may be different from the antibody-dependent cell cytotoxicity demonstrated in the peripheral blood leukocytes from volunteers who had a remote experience with influenza virus.


Interaction of glucosyltransferase with the cell surface of Streptococcus mutans. The partially purified glucosyltransferase (GTF) fraction synthesizing primarily water-insoluble glucans, GTF-A, and the homogeneous fraction synthesizing water-soluble glucans, GTF-B, were utilized to assess the binding of GTF activity to the cell surface of Streptococcus mutans GS-5. Growth of the cells in either Todd-Hewitt broth or a chemically defined medium did not appear to affect the ability of the cells to bind either enzyme fraction. Heat inactivation of the cells did not singificantly reduce the interaction of the enzymes with the cells. Cell surface glucan molecules appear to be involved in GTF binding to the cells because: (i) dextranase or alpha-1,3-glucanase treatment of the cells markedly reduced enzyme binding; (ii) the inclusion of soluble dextrans in the binding assays reduced both GTF-A and GTF-B binding to the cells; and (iii) pretreatment of the cells or the GTF-B fraction with soluble dextrans before binding significantly reduced enzyme binding to the cells. In addition, enzyme binding appears to require a cell surface protein component because Pronase, but not trypsin, treatment of cells reduced enzyme binding. Furthermore, the removal of a portion of the cell surface GTF-glucan complex with 3 N NaCl appears to provide additional binding sites for the enzymes. These results are interpreted in terms of the mechanism of the conversion of extracellular GTF to the cell-associated form.


Temperature-sensitive mutants of influenza A virus: production and characterization of A/Victoria/3/75-ts-1[E] recombinants. The Hong Kong/68-ts-1[E] virus, which has a 38 degrees C shutoff temperature for plaque formation, has been proposed as a donor of its two ts lesions to new variants of influenza A virus that pose an epidemic threat. To further examine whether the acquisition of the two ts-1[E] lesions will predictably attenuate new influenza A variants, the HK/68-ts-1[E] virus was mated with the A/Vic/3/75 wild-type virus. The Vic/75-ts-[E] recombinants that had the two ts-1[E] lesions also had a 38 degrees C shutoff temperature. Two Vic/75-ts-1[E] recombinants (clones 81 and 113) that had the two ts-1[E] lesions, a 38 degrees C shutoff temperature, and the Vic/75 hemagglutinin and neuraminidase glycoproteins were similar to each other and to their ts-1[E] parent in the pattern of replication and genetic stability in hamsters. These findings support the hypothesis that the acquisition of the two ts-1[E] lesions will predictably attenuate wild-type influenza A virus. Each Vic/75-ts-1[E] recombinant virus that possessed only the group 1 ts-1[E] lesion had a 39 degrees C shutoff temperature. Two of three of the Vic/75-ts-1[E] recombinants that had only the group 2 ts-1[E] lesion had a 39 degrees C shutoff temperature. This suggests that the HK/68-ts-1[E] donor virus contains two ts genes each of which by itself restricts plaque formation at 39 degrees C and above. The HK/68-ts-1[E] parent virus and its Vic/75 recombinant clones 81 and 113 were evaluated in ferret tracheal organ cultures maintained at permissive and restrictive temperatures. The Vic/75-ts-1[E] clone 81 differed from its parent and sister clone 113 in that it replicated readily and caused ciliostasis at 37 degrees C, a temperature restrictive for the replication of other ts-1[E] recombinants with a 38 degrees C shutoff temperature. The genetic basis underlying this difference was not elucidated.


Temperature-sensitive mutants of influenza A virus: evaluation of A/Victoria/3/75-ts-1[E] recombinant viruses in volunteers. The Hong Kong/68-ts-1[E] virus and its Udorn/72 and Georgia/74 recombinants, which have a 38 degrees C shutoff temperature and a ts lesion(s) on the genes coding for the P3 and NP proteins, were adequately attenuated and immunogenic in adult volunteers who lacked serum hemagglutination-inhibiting antibody (titer, </=1:8), but who possessed serum neuraminidase-inhibiting antibody. Two Victoria/75-ts-1[E] clones that also had a 38 degrees C shutoff temperature and a ts lesion(s) on the same two genes were administered to adult volunteers who lacked both serum hemagglutination-inhibiting antibody (titer, </=1:8) and neuraminidase-inhibiting antibody (titer, </=1:4). In contrast to the behavior of the earlier ts-1[E] recombinants, the Vic/75-ts-1[E] recombinants retained the capacity to cause febrile, systemic illness. However, the recombinants were attenuated compared with wild-type virus. The Vic/75-ts-1[E] virus vaccinees shed a larger amount of virus for a longer time than the previous ts-1[E] vaccinees, but they shed less virus than volunteers infected with wild-type virus. The ts-1[E] virus shed retained its ts phenotype in most instances and failed to spread to susceptible contacts. Vaccinees were partially protected against homologous wild-type virus challenge. The failure of HK/68, Udorn/72, and Georgia/74 ts-1[E] vaccinees to develop systemic reactions may reflect the presence of neuraminidase immunity before infection. In this situation, attenuation probably resulted from the degree of defectiveness of the ts-1[E] recombinant virus and the existence of neuraminidase immunity in the recipients. The 50% human infectious dose of the Vic/75 ts-1[E] virus was less than 10(5.2) 50% tissue culture infective doses. This suggests that at the time of a pandemic shift involving both the hemagglutinin and neuraminidase glycoproteins, a small amount of live virus vaccine might be effective in initiating infection.


Splenomegaly, enhanced phagocytosis, and anemia are thymus-dependent responses to malaria. Immunologically competent mice and mice with defined immunological deficiencies were infected with Plasmodium yoelii. Splenomegaly, enhanced phagocytosis, and anemia were most marked in infected mice having intact thymic tissue. Whereas the spleens of infected nude mice increased minimally in size, the relative blood hemoglobin levels and the rates of carbon clearance in these mice were similar to those of noninfected, immunologically intact mice. Thymus-reconstituted nude mice and B-cell-deficient mice responded to infection in a manner similar to that of infected immunocompetent mice. These data demonstrate that the hallmarks of malaria, i.e., splenomegaly, enhanced phagocytosis, and anemia, are thymus-dependent responses to infection.


Effect of cytochalasin D on Toxoplasma gondii cell entry. Cytochalasin D did not prevent attachment but did prevent entry of Toxoplasma gondii into peritoneal macrophages and bladder tumor 4934 cells. Inhibition of entry of T. gondii into peritoneal macrophages by cytochalasin D was dose related and comparable to inhibition of phagocytosis. Prevention of entry of T. gondii into bladder tumor 4934 cells by cytochalasin D followed a dose response identical to that observed with peritoneal macrophages. After removal of the medium containing cytochalasin D, its effect was completely reversible, and the kinetics of the loss of inhibition followed a similar time course for both phagocytic and "nonphagocytic" cell types. The studies support the concept that the host cells actively participate in the process by which T. gondii gains entry into cells.


Scanning electron microscopy of lymphoid cells from leukemia virus-infected mice. Spleen, lymph node, bone marrow, and thymus cells from Friend leukemia virus (FLV)-infected mice were examined by scanning electron microscopy. Whereas splenocytes from normal, noninfected animals showed the expected morphological classes of lymphocytes, including cells with numerous villous projections and smoother cell types, spleen cells from mice infected with FLV showed a rapid alteration of surface morphology. Shortly after infection, a decrease in the number and percentage of villous cells occurred, with a concomitant increase in the number of cells that were larger and smoother. Within 10 to 20 days after infection, the majority of splenocytes were smooth, large cells showing many distinct morphological charges, including surface "holes" and a "spongy" appearance. By days 25 to 35 after infection, most splenocytes were abnormal in appearance. Similar changes occurred in the lymph nodes after FLV infection, but the rate of change was much lower. Abnormal and larger smooth-surfaced cells did not become prominent until after week 2 or 3 infection. Thymus and bone marrow cells showed little if any change in surface morphology until late in the infectious process. However, even at that time only a few of the cells were abnormal in appearance. The changes in cell population in the spleen but not the lymph nodes paralleled the rapid decrease in the percentage of cells which stained positive for surface immunoglobulin and theta antigen. Futhermore, FLV antigen rapidly appeared on spleen cells after infection; fewer lymph node cells were positive, and only low numbers of marrow and thymus cells stained positive for FLV antigen. The marked immunosuppression induced by FLV infection paralleled and in some instances preceded the marked morphological changes.


Studies on a canine intestinal spirochete: scanning electron microscopy of canine colonic mucosa. A large homogeneous population of spirochetes was observed attached to the colonic mucosa of clinically normal dogs. Their association with the colonic mucosa and their mode of attachment resembled those of a spirochete reported in primates.


Toxicity of emulsified trehalose-6,6'-dimycolate (cord factor) in mice depends on size distribution of mineral oil droplets. Toxicity of emulsified trehalose-6,6'-dimycolate administered intravenously into mice depended on the size distribution of the mineral oil droplets. Emulsions containing the greatest number of the largest oil droplets were the most toxic. Emulsions of trehalose-6,6'-dimycolate made with peanut oil or with olive oil were less toxic than those made with mineral oil.


Application of charge shift electrophoresis to antigenic analysis of mycoplasmic membranes by two-dimensional (crossed) immunoelectrophoresis. Both the anodic migration and resolution of membrane antigens of Mycoplasma arginini were greatly improved in two-dimensional immunoelectrophoresis by incorporating both a neutral detergent, Triton X-100, and an anionic detergent, sodium deoxycholate, in the agarose support for first phase electrophoresis. The mobility of cytoplasmic antigens was unaffected at concentrations of 0.1% or less of sodium deoxycholate.


Suppression of Staphylococcus aureus extracellular nuclease and alpha-toxin synthesis by acetylmethylcarbinol. Acetylmethylcarbinol added to the inoculated culture of Staphylococcus aureus Wood 46 suppressed 85% of the differential rates of extracellular nuclease and alpha-toxin.


Sex hormones and the immune response. I. Host factors in the production of penicillin-specific antibodies in the female guinea pig. Female guinea pigs between 500 and 650 g produced the highest precipitation titers to penicilloyl-coupled guinea pig gamma-globulin of an array of animals ranging in weight from 350 to 850 g. When one or two depot injections in complete Freund's adjuvant were succeeded by saline boosters, the response maximized within 1 week after the first booster (in phase with the maximum output of IgM); subsequent boosters rapidly reduced the titer. The observed response was directed primarily at the hapten, as no reaction was evident to heterologous carrier (KLH) until the anti-hapten titer had declined. The daily administration of estradiol for 23 days after the last antigen inoculation prevented the decay of titer, whereas the titer of the controls or progesterone-treated animals dropped by 40--50% during that interval.


5-Hydroxytryptamine and rat passive peritoneal anaphylaxis. Antigen challenge of rats, sensitized by intraperitoneal injection with rat anti-serum, did not result in a detectable increase in the 5-hydroxytryptamine (5-HT) levels in their peritoneal fluids over the background level induced by sensitisation alone. The maximum amount of extravasation produced by intraperitoneal injection of 5-HT into passively sensitised rats was less than that produced by antigen or histamine, and the doses of 5-HT producing these levels of extravasation produced an produced an increase of 5-HT concentrations in the peritoneal fluids. Therefore, 5-HT is unlikely to make much direct contribution to the extravasation produced during rat passive peritoneal anaphylaxis. However, when given intraperitoneally to rats, 5-HT potentiates the extravasation produced by histamine.


Effect of antigen dose on the secondary IgE response in BN rats. Inbred Brown Norway rats were immunized at day 0 with a single dose of ovalbumin and 1 mg A1(OH)3. Various doses of ovalbumin without adjuvant were given at day 28. The secondary IgE antibody was antigen dose-dependent. Total serum IgE levels decreased after immunization and during the IgE antibody response.


The promise of the membrane artificial lung. Membrane lungs are more physiologic than bubble or disc blood oxygenators. Membrane lungs are indispensable in long term perfusions. Recently, there has been a noticeable trend toward use of the membrane lung in open heart surgery. Improper use of the membrane lung lowers its optimum potential and may lead to disenchantment on the part of the user. It is important to recognize that for optimum performance of the membrane lung it is necessary to learn to correctly use the membrane lung.


Morphologic examination of a prototype liver assist device composed of cultured cells and artificial capillaries. Cultures of the minimal deviation rat hepatoma cell line H4-11-E derived from the Reuber hepatoma were grown on bundles of artificial capillaries. Sections of those cells grown in circumfusion culture on acrylic copolymer and polysulfone capillaries were prepared. Light, transmission and scanning electron micrographs disclosed variations in cell viability in different parts of the bundle, variations in capillary wall structure affecting the proximity of cells to capillary lumen, and intraluminal sediment deposits derived from pump tubes used for circulating the culture medium. These sediments, by obstructing capillaries, inhibited cell growth and caused focal necrosis of cells. The importance of monitoring the morphology of cell cultures grown on bundles of artificial capillaries which are to be used as artificial organs is emphasized.


Response of malignant melanoma to plasma exchange, surgical debulking, and Corynebacterium parvum. A 48 year old man with extensive cutaneous and subcutaneous metastatic malignant melanoma responded to treatment by repeated plasma exchanges, partial surgical debulking of tumor masses, and intravenous Corynebacterium parvum. Circulating immune complexes were measured by the serum C1q binding method. Tumor regression coincided with serum C1q binding reduction to normal levels. Clinical relapse was preceeded by recurrent elevation of serum C1q binding activity.


A microcomputer based portable hemodialysis system. A microprocessor controlled portable hemodialysis system (The Suitcase Kidney) has been designed and 18 dialyses have been successfully completed. The microprocessor is shown to be an improvement and simplification over the conventional discrete control mechanisms presently used to monitor and control system functions. The microprocessor is shown to reduce development time, complexity and size of the system while improving patient safety. A description of the system is presented emphasizing the functioning of the microcomputer.


Continuous monitoring of urea and inorganic phosphate during hemodialysis: II. Clinical trials. Ion-selective eletrodes have been used to monitor urea and inorganic phosphate removal during dialysis. The dialysate outlow concentrations from a single pass beside controller were used to compute instantaneous serum concentrations. The urea data are in agreement with a single pool reservoir model; the inorganic phosphate removal was not log-linear, indicating rate limiting intracompartmental transfer. The procedure can be applied to the computation of urea generation rates and thus provide information on the protein catabolism rates of the patient; in addition, the monitor provides continuous checks on the dialyzer performance.


The conflict between anticoagulation and hemostasis during hemodialysis. Anticoagulation with comparatively small amounts of heparin has been carried out in more than 3,000 acute and chronic hemodialysis procedures without problems. In a selected high risk group of postoperative and polytrauma patients no patient hemorrhages have occurred, nor have there been any clotting problems in the dialyser circuit. Studies to elucidate the underlying mechanisms of coagulation and anticoagulation in the extracorporeal environment have been performed. Minimal intermittent heparin administration based on plastic anticoagulation monitoring with the APTT method, has proven to be particularly safe.


Haemoperfusion over ion exchange resins and polymeric adsorbents. A review of the development and application of haemoperfusion over ion exchange resins and polymeric adsorbents is presented. Aspects of biocompatibility and ion balance are discussed.


Recent changes in the morphology of gastric cancer in Japan. The macroscopical and histological features of 4,428 cases of gastric cancer resected stomachs was noted.d surgically between 1953 and 1974 were studied. Of these cases, 81.9% were in an advanced stage and 18.1% in an early stage. The following changes were observed. (1) In the group of advanced cancers, a gradual decrease in the frequency of Borrmann II type was noted. Most of these tumors were well-differentiated adenocarcinomas. (2) In the group of early cancers, a gradual increase in the proportion of "cancerous erosion" was detected. The majority of these tumors were poorly differentiated adenocarcinomas or infiltrating signet-ring cell cancers. (3) At intervals of 10 years, 5-10% decrease in the frequency of intestinal metaplasia in resected stomachs was noted.


The effect of mass screening in Iceland, 1965-74, on the incidence and mortality of cervical carcinoma. A clinic for early detection of cancer of the uterine cervix has been in operation in Iceland since 1964, aimed until recently at the age-group 25-59. More than 85% of women in this age group have been screened at least once. Mortality from cancer of the cervix had been rising in Iceland, and continued to rise during the first few years of operation of the screening clinic. Since 1970, however, a more than two-fold reduction in mortality has been observed among women aged 25-59. There has been a similar decrease in incidence of tumours of stages II, III and IV. Both deaths and advanced tumours are largely confined to women who have never been screened. Alternative explanations are considered, but the only tenable explanation of the reduction in mortality is that it is a consequence of the introduction of a comprehensive screening programme.


Melphalan-induced chromosome damage in sensitive and resistant human melanoma cell lines. Twelve consecutive treatments of a human melanoma cell line (MM253) with melphalan gave a subline (MM253-12M) which was five times more resistant to melphalan with respect to survival. In contrast to mustard-resistant rodent cells, the MM253-12M line had a higher stemline number than the parent line while growth rate and cell and colony morphology were unchanged. A further melphalan treatment following attempted mutagenesis with UV did not increase resistance. In a comparison of these two lines with two melanoma lines derived from other patients and the rat XC line, resistance was correlated with lower frequency of melphalan-induced chromosome aberrations, determined 48 h after a 4-h exposure to melphalan (3 microgram/ml). In the two cell lines studied, aberration-free metaphase cells from treated culture had fewer chromosomes than untreated cells. DNA synthesis studied in the 4- to 72-h period after treatment was inhibited to the same extent in MM253 and MM253-12M cells at 4 microgram/ml but to a greater extent in the sensitive line at 0.1-1.5 microgram/ml. During the first hour of treatment at 0.1-1.5 microgram/ml, DNA synthesis in MM253 appeared to be enhanced.