On the mechanism of action of biguanides. evidence against an inhibition of gluconeogenesis from protein in normal fasting subjects. The effect of phenformin on fuel homeostasis and on gluconeogenesis from protein was studied in 8 normal subjects who underwent two 4-day fasts 8-12 weeks apart. Each person received placebo or phenformin 50 mg every 12 hours for 3 days before and during the fasts. Circulating glucose, lactate, pyruvate, free fatty acid and ketone levels as well as urinary nitrogen excretion observed during placebo and phenformin treatment were not significantly different from each other. The lack of an effect of phenformin on urinary nitrogen excretion and plasma glucose level strongly suggests that gluconeogenesis from amino acids was unaltered by phenformin in these fasting subjects. The present findings are at variance with those by other authors which may be related to differences in methodology.


Pancreatic hormones and plasma glucose: regulation mechanisms in the goose under physiological conditions. I. Pancreatectomy and replacement therapy. Measurements of plasma GLI and IRI in normal fasting geese, before and during constant I.V. infusion of saline, gave GLI/I ratios of 1.32 +/- .07 and 1.34 +/- .03 (w/w). As total pancreatectomy markedly reduces the pancreatic hormone level, leading to a mortal hypoglycaemia, we attempted to maintain plasma glucose within the normal range by constant I.V. infusion of glucagon and insulin into operated animals. The results as follows: 1. Blood glucose levels can be maintained within the normal range during experiments lasting 6 or more hours with a constant G/I ratio. 2. The G/I ratio obtained in operated animals (.96 +/- .12) is near to, but significantly lower (p less than .005) than, the GLI/I ratio measured in normal animals. This difference may be explained by the presence of a small amount of circulating gut GLI in the 2nd group.


Effects of hexapeptide, a compound analogous to insulin B chain fragment B-21-26(DP-432) on the glucose uptake into the perfused hind limb of rats. DP-432 is a synthetic new peptide analogous to insulin B-chain fragment B21-26. It has been reported that this hexapeptide shows insulin potentiating action besides insulin-like activities in adipose tissue and diaphragm of rats. The perfusion of the hind limb of rats was performed according to the procedure of Ruderman with some modifications. The medium was Krebs-Ringer bicarbonate buffer containing 0.5% bovine albumin and 8.3 mM glucose. The erythrocytes were omitted from the medium. In this experimental procedure it was shown that the glucose uptake into the hind limb of rats increased by infusion of DP-432 (100 microgram/ml). The amount of glucose uptake into the hind limb responded to insulin doses (50, 100, 1000 muU/ml). DP-432 had insulin-like activity of 100 muU/ml. However, DP-432 itself did not have any potentiation of insulin activity on the muscle.


Increased glucose disappearance rate after short-term clofibrate administration in normal subjects and in patients with chemical diabetes. Following seven days clofibrate administration, the glucose disappearance rate during an intravenous glucose tolerance test was significantly increased in 6 normal subjects and in 11 patients with chemical diabetes. Both the acute and the total insulin secretory response were unchanged. Though fasting serum levels of free fatty acids, triglycerides, and cholesterol were significantly lowered by treatment, no correlation was found between the decrease in serum lipids and the increase in glucose disappearance rate. It is suggested and clofibrate improves glucose utilization by reducing insulin resistance irrespective of its effects on serum lipids concentration.


Epinephrine-sensitive adenylate cyclase of human fat cell ghosts. Modulation of enzyme activity by GTP. Some of the effects of GTP on human fat cell adenylate cyclase activity were studied. This nucleotide caused a dose-dependent inhibition of basal activity without affecting the hormone-activated rate of cAMP formation. Maximal effects were observed at GTP-concentrations of about 1 x 10(-4) M. The relative extent of hormonal stimulation was about 1.5-fold increased in the presence of 0.1 mM GTP.


Effects of fasting and refeeding on the in vitro insulin sensitivity of rat aorta. Rat aorta responds to refeeding after a fast in a manner similar to adopose tissue and liver by developing an enhanced capacity for lipogenesis and glycogen synthesis from glucose. The in vitro incorporation of D-U-14C-glucose into aortic triglycerides and glycogen was two- to four-fold higher in rats refed for three to five days after a three day fast than in ad libitum fed controls. Insulin significantly stimulated this incorporation only during refeeding for three days after a three-day fast. The glycogen synthesizing system appeared to be stimulated and to become sensitive to insulin earlier in the refeeding process than did the lipogenic system. The in vitro incorporation of 14C-glucose into aortic phospholipids was less affected by the nutritional state of the animal, and was not stimulated by insulin at any stage of the experiment. Possible mechanisms for the development of insulin supersensitivity and the implications for lipid accumulation in the artery wall are discussed.


Effect of alcohol on microsomal cortisol 4en-5 alpha-reductase in the liver of rats fed on a standard or low protein diet. Alcohol feeding (40% of total calories over a period of 9 days) increases microsomal cortisol-5 alpha-reductase activity in rat liver distinctly. It is assumed that this is an adaptive response to an increased release of cortisol caused by alcohol administration. Cortisol-5 alpha-reductase activity is decreased to one third of control values in rats fed an isocaloric, low protein diet. The response to alcohol feeding is susta ined in these animals. Phenobarbital treatment (80 mg/kg x day) stimulates 5 alpha-reduction of cortisol per g of microsomes almost twofold. The activity calculated per total liver increases 4-fold. Alcohol administration has no additional effect on cortisol-5alpha-reductase in phenobarbital-treated rats.


Characteristics of staff burnout in mental health settings. To determine the characteristics of staff burnout and ways of coping with it, the authors gathered data on institution-relation and personal variables for 76 staff members in various mental health facilities in the San Francisco area. A correlational analysis revealed a large number of statistically significant findings. For instance, the longer staff had worked in the mental health field, the less they liked working with patients, the less successful they felt with them, and the less humanistic were their attitudes toward mental illness. The authors present recommendations for reducing staff stress and subsequent burnout, including allowing more chances for temporary withdrawal from direct patient care and changing the function of staff meetings.


Using the Ward Atmosphere Scale to help change the treatment environment. The Moos Ward Atmosphere Scale was administered on a 45-bed psychiatric unit in a Veterans Administration Hospital to assess how staff, patients, and observers viewed the ward and to clarify what goals staff had and what changes they wished to make. Comparisons of staff's ratings of the existing milieu and of what an ideal milieu should be showed they wiched to improve the ward in the areas of patient involvement, support, spontaneity, order and organization, program clarity, and staff control; methods for making such changes became apparent. When the scale was administered again seven months later, respondents' ratings indicated that in general there had been substantial positive changes in all six areas. The authors believe the Moos scale is an invaluable aid in making and measuring change in a ward environment.


Attitudes of mental health center staff: changes over a five-year period. The author compares a study of attitudes of staff toward priorities and problems in eight community mental health centers with a study five years later, using the same questionnaire, of attitudes of staff in one of the centers. Between 1968 and 1973 the number of psychiatrists decreased and the number of psychologists and social workers increased. The most significant change in priorities in 1973 was the higher ranking of consultation and education services. While staff saw their problems in 1968 as a diffusion and inadequate program planning, by 1973 they were concerned with administrative problems and with shortcomings in the quality of clinical care, due to a lack of resources.


Staffing patterns and team composition.

Staffing patterns and team composition. The authors describe the work of a Veterans Administration task force in developing guidelines for staffing patterns and team composition on a hospital inpatient ward. The task force first defined the objectives of treatment, and then proceeded to define the needs of patients and their families, the types of professionals constituting the mental health team, and the functions they performed. It also identified the specific activities required in providing patient care, the staff involved in them, and those who could be involved, in some cases with more training.


Involving community representatives in CMHC evaluation and research. The participation of community representatives and consumers of services in agency planning has been mandated by legislation and the subsequent regulations. However, the representatives' lack of technical knowledge and sophistication frequently makes them unequal participants in the decision-making process. To help increase such sophistication, the program analysis and evaluation section of a large community mental health center has involved or assisted patient and community volunteers in various evaluation and research projects; they include a survey of area housing conditions, a survey of the needs of former inpatients, an evaluation of programs operated by a coalition of community groups, and a community resource listing.


Assessing community attitudes toward mental illness. Staff of a Veterans Administration hospital combined recruitment of family-care sponsors with an assessment of community attitudes toward mental illness. They mailed questionnaires to 989 persons in a small community 35 miles from the hospital. Most of the 235 respondents believed that mental illness is caused by a lack either of physical health or of proper nurturing, rather than seeing it as punishment for sins. Respondents were generally optimistic about the chances of recovery. Respondents with no children at home were more receptive to the ideal of accepting a family-care patient, while those with children at home were highly ambivalent. From the responses, the staff were able to locate several family-care sponsors.


Early planning helps hospitals run efficiently during strike. Careful planning can lessen the harmful effects of a strike on any hospital's day-to-day operations. A Baltimore hospital found that the use of contingency plan during an actual strike made the transition to a modified operational mode relatively smooth and that communication was enhanced. Because lines of responsibility and communication were clearly established, management was able to devote more time to negotiation and to making the contingency plan work rather than spending time on rush planning and policy interpretation.


Teaching hospital nurtures growth of community oncology clinics. In the belief that primary cancer care, such as drug and radiation therapy, can be delivered by community hospitals close to patients' homes and families, University Hospital and the Boston University School of Medicine are helping to establish local oncology clinics. Educational services for community physicians, nurses, and social workers are also offered.


Small firms look to hospitals for occupational health services. Some hospitals are offering preemployment physicals, routine examinations, prompt work injury care, physician and nurse consultations and visits, and other services to small companies that cannot justify establishing in-house occupational helath units.


Cost containment in laundry and linen service. One major problem looms among all others in the area of laundry and linen service, whether a hospital has an in-house operation, is part of a shared or central laundry, or uses a commercial service. That is the costly problem of controling linen consumption and replacement. A recent seminar offers some insight into reasons for the problem and some possible solutions.


Limited, periodic surveillance proves practical and effective. Little data exist as to which type of surveillance--continuous or periodic--is most effective against nosocomial infection. Study proved periodic data collection a simple, practical, and effective method for identifying and monitoring trends. More importantly, the system provided more time for the infection control nurse to spend on corrective measures.


Standards of performance developed for clinical dietitians. Although standards of performance are common to industry, they are relatively new to health care professions. The development of standards of performance for registered dietitians in a clinical capacity are described. Changes made after a one year review of the program are listed. The benefits and costs of the program also are explained in the article.


Emergency services: basic assessment model proves effective, adaptable, economical. A simple five-step quality assessment model has been easily and economically implemented in emergency departments of varying size and activity. The model has encouraged quick action in correcting deficiencies, systematic problem solving, and the development of ED standards and criteria.


Health care delivery: strength in numbers. The number and types of multi-institutional arrangements among health care facilities are on the increase, and the days of the completely autonomous, independent hospital are coming to a close. Although by themselves hospital systems are no panacea in dealing with the challenges facing hospitals today, many such arrangements offer more opportunities than problems in coping with the rapid changes currently facing the health care industry. The pros and cons of seven arrangements are discussed in detail.


System predicts patient census, forecasts staffing needs, costs. Given the present climate concerning hospital costs, it has become imperative for hospitals to take steps to ensure that they provide care in the most efficient possible manner. Donald N. Sharp Memorial Community Hospital has developed a system that allows it to respond with unusual precision to changing patient workloads. Both staffing and other operations can be altered to meet a changing patient census.


It takes a sense of survival.

It takes a sense of survival. There is a legitimate role for the small urban hospital. In order to perform that role optimally, however, the small hospital must select the services that it provides carefully, avoiding those that are better suited to larger institutions, and must investigate the appropriateness of sharing services with other small hospitals.


Decentralized pharmacist concept solves unit dose problems. In order to solve problems associated with the unit dose drug distribution system at LDS Hospital, the decentralized pharmacist concept was tried. The pharmacist uses a master medication cart to fill new drug orders at the nursing station and a computer video terminal at the nursing station to check patient profiles. The reactions of nurses and members of the medical staff to the decentralized pharmacist are described.


Energy conservation is everybody's business. The energy crisis is affecting all aspects of society, including hospitals and health care institutions. In keeping with the need to conserve energy, the food service administrator should consider implementing an energy management program. Various resources that can help the food service administrator to conserve energy are described herein.


In-house staff designs energy-saving system. With a relatively small amount of money and a lot of ingenuity, an innovative engineering team developed a successful pilot program that resulted in substantial energy and dollar savings. Using commercially available component parts, the team designed a preventive maintenance and energy-saving microprocessor system that is especially geared toward the hospital setting.


We have seen rate review, and it works.

We have seen rate review, and it works. In Maryland, state rate review is successful because of cooperation at all levels. Although there was friction in the beginning between the state commission and health care providers, the latter, with strong encouragement from their associations, played an important role in adopting a constructive program.


Hospital philanthropy faces uncertain future. Two pieces of legislation proposed by the Carter Administration may result in dire consequences for hospital philanthropy, experts fear. In an effort to alert health care executives as to how these laws and the possible demise of charitable giving could harm their institutions--and the future of this country's health care in general--the National Association for Hospital Development recently held a governmental affairs conference in Washington, DC. The highlights of that meeting are presented herein.


Human values vs. political and economic realities. There is no question, as in other industries, that there is room for improvement in hospitals' performance. However, the managers of these institutions can be justifiably proud of their past performance. There is little evidence that further intrusion in health care matters by the federal government will lead to significant reductions in the cost of health care services. The evidence seems to suggest that health care providers, working together, can have such an effect.


Five groups start community cardiac rehabilitation program. Five local organizations jointly developed a cardiac rehabilitation program for the community of Cedar Rapids, IA. The program, which is designed as a vehicle to improve the quality of life for persons with heart disease and to retard the further development of coronary artery disease, includes a preliminary patient examination, a planned exercise regimen, and an educational component, covering various aspects of heart care.


Conflict of interest and the banker-trustee. While hospitals benefit from banker board members' advice, they need to avoid the potential conflict-of-interest situations arising from financial decision making that may involve such members' institutions. Two effective ways of doing this are developing directives to be carried out by the hospital's financial manager and stimulating competition among financial institutions for the hospital's business.


The AHA cost and intensity indexes: a rejoinder. Conventional studies point to the spiraling cost of hospital services as the principal, if not the only, cause of escalating hospital expenditures; but using the AHA's Hospital Intensity Index to obtain a measure of the "real" volume of services provided by the industry opens up a whole new perspective--a perspective that gives attention to the role played by increased demand for services, as opposed to mere cost control.


Marketing ambulatory care.

Marketing ambulatory care. A number of environmental factors are bringing about the "day of the vertical patient"--making the providing of comprehensive ambulatory care important for a hospital's survival. Careful design and implementation of ambulatory care programs must be accompanied by equally well-considered marketing strategies.


Hospital care for the terminally ill: a model program. While community hospitals increasingly are becoming community health care centers, evidence suggests a great need for most of these institutions to improve their care of the terminally ill. Based on a study of existing care programs and of thanatology literature, the authors have developed a model hospital program for dying patients and their families that uses a team approach to integrate resources for their care.


Praying with the terminally ill.

Praying with the terminally ill. Terminally ill persons and their families will communicate their own prayer needs to healing persons who are attending carefully. A number of guidelines may also be helpful to healers in developing the personal characteristics needed to minister effectively and in determining when and how to pray with patients.


Peer review of hospital social work.

Peer review of hospital social work. To comply with PSRO legislation, St. Vincent's Hospital and Medical Center of New York (NYC) in 1977 established a social service peer review program. Detailed criteria are used in evaluating completed cases from 10 social service staff members each month, with the whole social work staff participating as evaluators on a rotation basis. The hospital expects the ongoing review program to benefit its social service practice in many ways and ultimately to upgrade patient care.


Selecting a CPA firm.

Selecting a CPA firm. Careful selection of a public accounting firm involves the hospital's administrators in a sensitive process. Attention to identifying prospective firms, applying selection criteria, requesting proposals, and preparing for the firms' presentations enables the administration to choose wisely.


Assessing trends in perinatology.

Assessing trends in perinatology. Hospital administrators must respond as the medical profession renews its efforts to reduce newborn mortality rates. Technological changes in the case of obstetric and newborn patients, pressures for the regionalization of service, and consumer demands for a more humane birth environment are factors the administrator must consider.


A Catholic hospital and natural family planning. Instituting a natural family planning program in a hospital setting is a challenge that the Catholic medical community is bound to accept. Natural planning methods, taught by appropriate and well-trained instructors, can help clients become more conscious of their fertility and more able to accept responsibility for their life decisions.


Early nutrition: its long-term role.

Early nutrition: its long-term role. The widespread belief that problems such as lifelong obesity originate in infancy sometimes leads to overzealous attempts to regulate energy intake too soon and may compromise linear growth. Treatment of obesity should wait unitl after age one.


Immunopathology of rheumatoid arthritis. The central place of immunologic aberrations in the pathology of rheumatoid arthritis is now well established. It is the nature and interactions of these aberrations that still remain to be elucidated. Among the many puzzles being studied are the uniquely self-perpetuating nature of rheumatoid arthritis immune complexes and the role of complement and the prostaglandins in joint damage and pain.


Managing the solitary thyroid nodule: echography's role. Improved preoperative thyroid diagnosis and reduction of needless thyroid surgery are achieved by employing gray scale echography in patients whose solitary thyroid nodule is cold on scintillation scanning. It is highly useful in distinguishing solid tissue from a cyst, identifying degenerative changes, guiding needle biopsy, and monitoring changes in a nodule during suppressive therapy.


Clinical pharmacology and toxicology of antidepressants. Described as the drugs of choice in treating acute depression, the tricyclics are unusual in that too high a dosage may be as ineffective as one that is too low, and plasma levels may vary as much as 30-fold among patients given the same dosage. After the recommended approach to determining appropriate dosage has been tried, the 15% or so of patients who do not respond are not likely to benefit from a switch of drugs.


Mucocele-like changes in Bartholin's glands. Two cases are described in which changes identical to those of oral mucocele occurred in Bartholin's glands. The histopathology, histochemistry, and pathogenesis are discussed.


The circulating "atypical" lymphocyte.

The circulating "atypical" lymphocyte. Atypical lymphocytes have been observed in the peripheral blood of patients in a large number of clinical situations, including immune reactions to transplantation and immunization, collagen diseases and other autoimmune disorders, malignant disease, drug reactions, and infectious mononucleosis, as well as other bacterial and viral infections. These cells are readily identified by their increased size and the presence of active DNA synthesis. In morphology, they closely resemble lymphocytes transformed into blasts by exposure to mitogens or antigens in vitro. They vary in morphologic detail as well as surface marker characteristics, indicating that they comprise a heterogeneous mixture of cell types. These data suggest that atypical lymphocytes may represent a polyclonal immune response to antigenic stimulation.


A fetus with recombinant of chromosome 8 inherited from her carrier father. A pericentric inversion of chromosome 8, inv(8)(p23q22), in a male carrier resulted in an unbalanced recombinant, rec(8)dup q, inv(8)(p23q22), which was diagnosed prenatally. The features seen in the aborted fetus resembled the features seen in a previously affected child who received the identical recombinant from her carrier mother. In this particular inversion involving chromosome 8, both male and female carriers risk producing an unbalanced progeny. Different familial pericentric inversions are reviewed for the presence or absence of unbalanced recombinants.


X-X translocation in a patient with gonadal dysgenesis and the problems of phenotype-karyotype correlations. A sex-chromatin-positive woman without stunted growth, but with primary amenorrhea, and some stigmas of pure gonadal dysgenesis had the chromosome constitution 45,X/46,Xt(X;X)(q27;q27). The abnormal chromosome formed a large Barr body and was late-labeling. The chromosome consisted of two X chromosomes attached by their long arms (end-to-end), both apparently having the partial distal deletion. Both centromeric regions showed C-staining but only one constriction. The chromosome is interpreted as an isodicentric with only one centromere functioning. Some problems of phenotype-karyotype correlations are discussed.


Lysosomal enzyme activities in different types of amniotic fluid cells measured by microchemical methods, combined with interference microscopy. In primary amniotic fluid cultures, four distinct types of cells were characterized as epithelioid (E I and E II), fibroblast-like (F), And large cells, Small numbers (1-200) of freeze-dried cells were isolated from colonies of each cell type and analyzed for the activity of three lysosomal enzymes: beta-N-acetylglucosaminidase, beta-galactosidase, and alpha-glucosidase. When expressed per cell, the activities for each of the enzymes were not significantly different among the small types of cells (EI, EII, and F). However, 5 to 10-fold higher enzyme activities were found in the large cells. The dry mass of individual large cells, as measured by microinterferometry, was also 5 to 10 times higher than that of the smaller cell types. When expressed per unit of dry mass, the enzyme activities tested, appeared to be independent of the type of amniotic fluid cell. The significance of this observation for the rapid prenatal diagnosis of metabolic diseases is discussed.


Molecular and functional anomalies in two new mutant glucose-phosphate-insomerase variants with enzyme deficiency and chronic hemolysis. Two new deficient glucose-phosphate-isomerase (GPI) variants have been described in patients suffering from severe chronic hemolytic anemias. The patients' parents were consanguineous, such that the patients were true homozygotes for the mutated GPI genes. In both cases the main cause of the defect in enzyme activity was molecular instability of the mutated GPI molecules, their catalytic activity being nearly normal. GPI 'Paris' was characterized by a slow electrophoretic migration and, above all, a drastically altered affinity for the substrates glucose-6-phosphate (decreased) and fructose-6-phosphate (increased). GPI 'Enfants malades' exhibited a slightly reduced electrophoretic mobility, an abnormal curve of the activity in function of pH, and an abnormal ratio of maximal velocity in the backward direction (fructose-6-phosphate leads to glucose-6-phosphate) to that in the forward direction (glucose-6-phosphate leads to fructose-6-phosphate). No clear relation could be proved between the kinetic abnormalities of the mutant GPI variants on the one hand and the metabolic changes of the GPI-deficient red cells and the severity of hemolysis on the other. Finally we emphasized the possible role of the impairment of hexosemonophosphate pathway in the reduction of viability of the GPI-deficient red cells.


A new variant of human erythrocyte G6PD occurring at a high frequency amongst the population of two villages in The Gambia, West Africa. During the course of a large survey of red cell G6PD genotypes in The Gambia, a slow electrophoretic variant with reduced enzyme activity was found to occur at a high frequency. This variant, G6PD Gambia, was found in the following genotypic combinations: males; G6PDGam, females; G6PDA+/Gam, G6PDB+/Gam, and G6PDA-/Gam. From the electrophoretic mobility and kinetic characteristics it was concluded that G6PD Gambia was a hitherto unreported variant of G6PD. The frequency of the G6PDGam gene amongst the 1109 individuals examined was 0.024.


Geroderma osteodysplastica. A report of two affected families. This paper describes two families in which four boys and two girls were affected with geroderma osteodysplastica. The major features of this syndrome include a droopy, jowly, prematurely aged appearance that has been likened by previous authors to the dwarfs in Walt Disney's 'Snow White.' Also, their skin lacks normal elastic recoil. The bones are osteoporotic and susceptible to fractures, particularly the vertebrae, which show compression with anterior wedging and biconcavity. The findings in the present families are compared with those in the originally reported family. There was a high degree of consanguinity between the parents in both of the present families and the previous hypothesis of X-linked inheritance is questioned.


t(9/22) with centric fission and NOR translocation leading to a case of pure 9p trisomy in the offspring. A case of trisomy 9p originating from t(9/22) mat is reported. The rearrangement is characterized by centric fission of chromosome 9 and by translocation of NOR from chromosome 22 to 9q. The hypothesis of centric fission is discussed on the basis of various banding patterns. The case also reveals unusual knee dislocation.


Partial trisomy for the long arm of chromosome 3 [3(q21 to qter)+] in a newborn with minor physical stigmata. In this report a description is given of a female newborn with minor physical stigmata presenting a partial trisomy for the second region of chromosome 3 (3q32 leads to 3qter). The additional 3q2 material is located at the distal end of the long arms of chromosome 2 as the result of a de novo 3/2 germinal translocation.


The Roberts' syndrome.

The Roberts' syndrome. A report is given on a small-for-date male infant showing the following symptoms: bilateral aplasia of humerus, radius, and ulna, shortened femora, bilateral cleft lip and cleft palate, stigmata of dysmorphism, and notably; simple helix formation of the ear, simian crease, clinodactylia, bilateral clubfoot deformity, hypospadia, thrombocytopenia, micrognathia, and contractures in the knee joints. Postmortem autopsy revealed horsehoe kidney, ureterstenosis with hydronephrosis, persistent branchial arches, and absence of the knee joints. Chromosome analysis results performed by G-band technique turned out normal. This, obviously, was a case of the so-called Roberts' syndrome. Our results were compared with the relevant literature and some particularities were emphasized. The question was discussed as to whether the SC-phocomelia (pseudothalidomid syndrome), the TAR syndrome, and reported single cases might be an identical syndrome.


Balanced t(8;9)(q12;q33)pat carrier with phenotypic abnormalities attributable to a de novo terminal deletion of the long arm of chromosome 7. A girl observed from birth to age 16 months had multiple congenital anomalies including growth and developemental retardation, microcephaly (-4SD), bulbous nose, prominent lips and philtrum, esotropia, latent hypermetropia, and spasticity. Chromosome analysis showed her to be a balanced carrier of a t(8;9)(q12;q33)pat translocation. In addition, she had a de novo deletion of a distal segment of the long arm of chromosome 7. Seven previously reported cases with deletions involving 7q were reviewed and had a number of nonspecific features in common, with microcephaly of a comparable degree in one of these. Studies of the Kidd (Jk) blood groups and Hageman factor were done because of the tentative assignment of their respective loci to distal 7q. Location of the Kidd (Jk) locus on the deleted segment can be excluded on the basis of heterozygosity of the proposita for the a and b alleles. Hageman factor was not decreased, which suggests that this locus is also not on the deleted segment.


Variant chromosome 3 (inv3) in normal newborns and their parents, and in children with mental retardation. The chromosomes of 102 normal newborn babies (51 boys and 51 girls) born at term, their parents, and 45 nonrelated children with mental retardation at the level of imbecility were investigated by fluorescence microscopy using propyl quinacrine mustard. In each of the 11 families, one of the parents had a variant chromosome 3 that was interpreted as resulting from a pericentric inversion of the brilliant band (or C band) only (inv3). In four cases inv3 was transmitted to the child. The frequency of inv3 in newborn boys and girls was 2 and 6% respectively, and in adult men and women 5 and 6% respectively. In children with mental retardation of unknown etiology the inv3 was detected in five cases (11.1%). This difference from normal persons was not significant.


Trisomy 10p due to t(5;10)(p15;p11) segregating in a large sibship. A family is reported with a segregating t(5;10)(p15;p11) translocation resulting in a child carrying trisomy 10p. The clinical findings of the patient are compared with trisomy 10p and the Cri-du-Chat syndrome.


Terminal deletion of (1)(q42) and its phenotypical manifestations. A 5-year-old male with multiple malformations (dwarfism, microcephalia with brachycephalic shape of skull, mongoloid lid axis, epicanthus, convergent strabismus, flat root of the nose, micrognathia, missing uvula, deformed low-set ears, hypoplastic genitals, and general hypotonia), severe mental retardation, and cerebral paroxysms caused by a partial monosomy (1)(q42 leads to qter) is described. This case is compared with other cases with a partial monosomy or ring-1 chromosomes.


Patterns of association in the human metaphase complement: ring analysis and estimation of associativity of specific chromosome regions. The pattern of metaphase chromosome association in the human complement was studied by two methods of statistical analysis of interchromosomal distances. Those methods included ring analysis in which a characteristic position of the centromere of each chromosome relative to the center of a two dimensional representation of a metaphase complement was defined, and estimation of the capacity for associativity of each of three regions of each chromosome: the centromere (c) and the ends of each arm (p, q). The following information was obtained: 1. In general, the distance from the center is directly related to chromosome size. 2. The most notable deviation from that size-related progression is displayed by the X chromosomes. The markedly peripheral position of the X is characteristic of both X's of the female and the single X of the male. 3. The relative associativity of each chromosome of the complement is, in general, inversely related to size with an additional preferential capacity of associativity displayed by the acrocentric chromosomes. Analyses of the different inter-regional classes established that the supplementary associativity factor of the acrocentric chromosomes was inherent in their pericentromeric and p-arm regions and excluded the ends of the q arms from participation in that factor. 4. Those analyses demonstrated that the specific morphology or 'geometry' of the acrocentric chromosomes contributes little to their high relative associativity. In addition to the tendency for the c/p regions of the acrocentric chromosomes to associate with each other, presumably because of their common function in nucleolar organization, those regions also displayed a propensity to associate with the distal regions of the arms of other chromosomes. A molecular basis for that propensity other than that of ribosomal DNA is postulated to be that of other fractions of highly reiterated DNA sequences. 5. Analysis of the relative associativities of each of the three regions of the Y chromosome revealed that the Yq displays a much stronger capacity to associate with the c's of other chromosomes than does the Yc or Yp.


Identification of the origin of triploidy by HLA markers. HLA-A and HLA-B markers have been determined in fibroblasts grown from tissues of triploid conceptuses and have been tested in the parents. Informative data on the origin of triploidy were obtained in eight cases: diandry I or dispermy in 4 cases, diandry II or dispermy in 2, digyny I or II in 2. This confirms that triploidy involved more frequently two sets of paternal chromosomes.


Reduced frequencies of mitomycin-C induced sister chromatid exchanges in AKR mice. The frequencies of base-line and Mitomycin-C (MMC) induced sister chromatid exchanges (SCE) were surveyed in four inbred strains of mice. In contrast to the C57Bl/6J, CBA/J, and A/J strains where frequencies of SCE increased linearly with increasing dose of MMC, levels of SCE were significantly lower in AKR/J mice at high MMC concentrations. At a dose of 5 mg/kg MMC, chromosomal aberrations were more frequent in bone marrow cells of AKR/J mice than in C57Bl/6J mice. These observations suggest an altered response to DNA damage in the AKR mouse strain.


Human erythrocyte pyruvate kinase deficiency: the use of a kinetic study of mutant enzymes for the detection of heterozygotes. Erythrocyte pyruvate kinase (PK) deficiency was detected in a boy of dutch origin. Immunologic, electrophoretic, and kinetic studies of the enzymes of propositus and the members of his family demonstrated that the boy was heterozygote for two different mutant PK alleles. The mutant enzyme, for which his mother was heterozygote, was characterized by a lower immunologic specific activity, a decreased affinity for the substrate phospho-enol-pyruvate, and a loss of homotropic interactions toward this substrate, an increased affinity toward the allosteric inhibitor MgATP2-, a decreased affinity for the activator fructose-1,6-diphosphate, and a lowered pH optimum. Electrophoresis, Km app. for MgADP, and the reactivity toward the purine nucleotide substrate analogues were normal. The mutant enzyme for which his father was heterozygote was characterized by a decreased affinity for the substrate phospho-enol-pyruvate and a loss of homotropic interactions toward this substrate. All other parameters mentioned above were normal. The use of a kinetic study of mutant enzymes for the detection of heterozygotes is discussed.


HLA antigen, gene, and haplotype frequencies in Thailand. Antigen, gene, and haplotype frequencies as well as phenotype distribution of the HLA system were studied in a series of 213 individuals in northern Thailand. The series consisted of 160 northern Thais, 23 Thai individuals from various other regions of Thailand, and 25 persons of Chinese  origin. Most frequently found were the alleles HLA-A11 and HLA-Bw40 and the haplotype HLA-A2,B-. Phenotype distribution followed a Hardy-Weinberg expectation. Significant differences were found especially between our results for the alleles of locus B and the results of a series from Bangkok reported by Chiewsilp and Chanarat (1976).


An increased frequency of chromosomal changes and SCE's in cultured blood lymphocytes of 12 subjects vaccinated against smallpox. An increased frequency of chromosomal changes and sister chromatid exchanges was detected in 10 women 7 days after smallpox vaccination.


Occurrence of 1st division metaphases in human lymphocyte cultures. Human lymphocytes were cultured for 40, 42 and 44 h and analysed for the number of 1st and 2nd division metaphases using the Brd U-Giemsa technique. The most suitable time to obtain the majority of cells in their 1st division concommittant with an adequate mitotic index was 42 h. It is recommended that laboratories use sister chromatid differential staining to determine the most suitable culture time for the analysis of 1st division metaphases. This will permit accurate comparisons between individuals and laboratories to be made.


Expression of IgGFc receptors on human plasma cells. Plasma cells that were obtained in bone marrow aspirates from patients with plasma cell dyscrasias were assessed for the expression of IgGFc receptors. A sensitive EA rosette assay and direct immunofluorescence were combined to quantitate plasma cells that expressed IgGFc receptors. Two patients with nonsecretory disorders had large numbers of receptor positive cells (98 and 78%). The remaining patients, all of which had been treated with cytotoxic drugs prior to testing, had significant numbers of IgGFc receptor positive cells (36-86%) and the number of FC receptor positive cells was increased by storing the cells at 4 degrees C for 18 hours prior to performing the rosette assay.


Preparative immunoabsorption electrophoresis. A method for separating a component from a mixture of antigens is described. The component which may be a virus or subfraction of a virus, is isolated by driving the mixture by electrophoresis through a gel containing precipitating antibodies directed against the unwanted components. The method is illustrated by the isolation of hepatitis B antigen from whole serum and by the separation of wild cucumber mosaic virus from a strain of tobacco mosaic virus.


The persistent PHA-responsive population in the mouse thymus. I. Characterization of the population. Using an in vitro culture technique, mouse thymus graft cells were co-cultured with peripheral blood lymphocytes in the presence of phytohaemagglutinin (PHA). The persistent PHA-responsive thymus graft population (Elliott, 1973) was shown to be able to respond to other T-cell mitogens (Con A, pokeweed mitogen, staphylococcal enterotoxin B), but not to E. coli lipopolysaccharide a known B-cell mitogen. The percentage of persistent PHA-responsive cells did not alter during 5 days in culture and was relatively unaffected by either hydrocortisone or anti-lymphocyte serum treatment in vivo. In allogeneic thymus grafts (AKR leads to CBA), persistent PHA-responsive cells could be demonstrated, which were destroyed when incubated with CBA anti thetaAKR serum and complement. When thymus graft cells were allowed to sediment on a 0.2--2 per cent BSA gradient, it was seen that the PHA-responsive population sedimented faster than the bulk of thymus graft cells. Some separation could be obtained on this gradient between the persistent and non-persistent PHA-responsive cell populations.


Acquired resistance to ticks. III. Cobra venom factor and the resistance response. Guinea-pigs developed resistance to larvae of the ixodid tick, Dermacentor andersoni, after one infestation. Resistant hosts were characterized by allowing significantly fewer larvae to engorge than non-resistant hosts. Larvae engorging on non-resistant hosts had a mean weight six times that of larvae obtained from resistant hosts at the end of a 5-day infestation. This immunologically based resistance was previously shown to have a cell-mediated and a humoral component. In an attempt to ascertain the role of complement in the resistance response, cobra venom factor (CoF) was administered to guinea-pigs producing prolonged (85--95 per cent) depletion of complement titres. CoF was administered during an initial infestation with tick larvae to determine if complement depletion altered the acquisition of tick resistance. CoF was also administered to tick-resistant hosts in an attempt to determine if the expression of tick resistance and the development of the basophil-packed lesion, characteristic of the tick-attachment site in resistant hosts, could be altered by complement depletion. CoF did not alter the acquisition of resistance when complement levels were reduced during a primary infestation. However, complement depletion of an animal which had acquired tick resistance blocked the expression of that resistance during a challenge infestation. In addition to increased numbers and weights of larvae engorging on tick-resistant animals depleted of complement, the basophil packed lesion at the tick attachment site was greatly reduced. Complement plays an important role in the expression of tick resistance in guinea-pigs.


Homocytotropic response to guinea-pigs to antigen extracts from formalized and unformalized Parascaris equorum. A homocytotropic anaphylactic response is described in guinea-pigs sensitized with antigen from Parascaris equorum. This antigen was not sensitive to prolonged storage in formalin. Some adult ascarid extracts prepared from worms that had been stored for 14 days in 10 per cent formalin before use, retained their ability to produce an allergenic response. The antisera withstood heat inactivation at 56 degrees for up to 7 h or treatment with 2-mercaptoethanol and were still capable of eliciting a skin response. The skin response persisted for 96 h after sensitization. The serum of animals sensitized with a single dose of P. equorum antigen remained capable of producing a positive passive cutaneous anaphylactic response up to 245 days after the sensitizing dose. It appears, from the results obtained, that this antibody resembles IgG1, rather than IgE.


Lipid A-induced tolerance and hyperreactivity to hypothermia in mice. Mice responded to lipopolysaccharide (LPS) with a dose-dependent, monophasic hypothermia reaching a maximum at 2 h postinjection. Degraded polysaccharide was not active; free lipid A, however, induced a similar pattern of hypothermia, indicating that the hypothermic principle of LPS was embedded within the lipid A component. The hypothermic response of mice to LPS was modified by prior exposure of the host to LPS. This altered reactivity was manifested by refractory periods (early and late tolerance), in which animals no longer responded with hypothermia, or a hyperreactive phase (hypersensitivity), in which hypothermic responses were greatly augmented upon LPS challenge. Thus, tolerance observed 24 h after a single injection of LPS (early tolerance) was followed, on further LPS challenge, by an enhanced hypothermic responses reaching a maximum on day 4. Further daily exposure of the animals to LPS eliminated hyperreactivity and led to the establishment of a late tolerance maximally expressed on day 8. Hyperreactivity could also be evoked on day 4 after a single injection of LPS. Mice pretreated with Salmonella S- and R-form LPS or free lipid A (Salmonella) demonstrated tolerance and hyperreactivity to both homologous and heterologous challenge. In addition, complete cross-tolerance was observed with S-form LPS derived from Shigella. It was concluded that the differential effects of LPS on host responses (tolerance and hyperreactivity) were due to lipid A.


Antibody responses in patients with rubella infection determined by passive hemagglutination, hemagglutination inhibition, complement fixation, and solid-phase radioimmunoassay tests. Antibody responses in serial serum specimens collected from 31 patients with an acute rubella infection were determined by passive hemagglutination (PHA), hemagglutination inhibition (HI), complement fixation (CF), radioimmunoassay (RIA) immunoglobulin G (IgG), and RIA immunoglobulin M (IgM) tests to evaluate the effectiveness of these tests in diagnosing a recent infection. The HI, RIA IgG, and RIA IgM antibodies appeared almost simultaneously and reached the maximum level about 1 week after the onset of rash. Compared to these, the CF antibodies developed only slightly later, whereas the development of the PHA antibodies was much more delayed. The RIA IgM response was shown to be transient, lasting approximately 1.5 to 2.5 months postinfection. The results of this study indicate that demonstration of specific IgM antibodies is the best method for diagnosing a recent infection, one within 2 months after the onset of the illness. If an IgM test is not available, a combination of the HI and PHA tests is recommended.


Microflora analysis of a child with severe combined immune deficiency. An infant with severe combined immune deficiency was delivered by Caesarean section and continuously maintained in an isolator in an effort to restrict microbial contact. However, during the past 5 years the child came in contact with at least 54 different microbial contaminants. His skin autoflora was similar to the reference group of healthy male adults in numbers of different species and the number of viable cells present per square centimeter of surface area. The subject's autoflora differed from the reference group in that significantly fewer anaerobic species were recovered from the patient's mouth and feces. The populations on the skin and in the upper respiratory tract were almost exclusively composed of staphylococci and micrococci, although several species that commonly inhabit these areas were infrequently isolated and failed to colonize. The fecal autoflora was dominated by staphylococci, and bacilli clostridia recovered in typical numbers. Enterobacter agglomerans, not commonly recovered from the reference group, was a frequent inhabitant of the mouth and feces of the child. The fact that this severe combined immune-deficient patient has remained disease free in the presence of this microbial population illustrates the noninvasive nature of the bacteria present, as well as the importance of unaffected components of the child's immune defense mechanism.


Use of Semliki forest virus to identify lipid-mediated antiviral activity and anti-alphavirus immunoglobulin A in human milk. In a simple and reliable assay system, Semliki forest virus (SFV) was used to detect the activity of antiviral factors in human milk. Fractionation of the  milk showed that a heat-stable, lipid-associated activity and an immunoglobulin-associated activity were present, either singly or together, in 85% of the human milk samples tested. Cow and synthetic milk showed neither activity. Extraction of the neutral milk lipids allowed the antiviral to be located with the monoglyceride and free fatty acid fractions. The milk low in antiviral lipids and high in triglycerides also lacked a strong lipase activity. The immunoglobulin anti-SFV activity was shown to be due to immunoglobulin A, the major milk immunoglobulin, and appears to be directed against an alphavirus closely related to SFV, possibly Ross River virus.


Cellular adherence, glucosyltransferase adsorption, and glucan synthesis of Streptococcus mutans AHT mutants. Streptococcus mutans AHT mutants M1, M2, and M13 failed to adhere to a glass surface, whereas mutants M9 and M35 exhibited decreased and increased adherence, respectively, as compared with the parent strain, when grown in sucrose broth. Extracellular glucosyltransferase prepared from glucose-grown cultures of the adherent strains (wild type, M9, and M35) induced adherence of heat-killed cells of the homologous and heterologous streptococcal strains as well as of Escherichia coli K-12 and uncoated resin particles. The glucosyltransferase was adsorbed on all the streptococcal cells and glucan-coated resins, but not on E. coli cells and the uncoated resins. Glucosyltransferase from the nonadhering mutants (M1, M2, M13) neither was significantly adsorbed on nor induced adherence of any of the cells and resins. Cell-free enzymes from the glucose-grown adherent strains produced water-soluble and water-insoluble glucans, whereas those from the nonadhering mutants produced only water-soluble glucans. Small amounts of alkali-soluble, cell-associated glucan were recovered from the sucrose-grown nonadhering mutants. Thus, the relative proportions of glucosyltransferase isozymes elaborated by the S. mutans mutants, insofar as they affect the physico-chemical properties of the glucans produced, seem to determine the adherence abilities of the cells. The adsorption of glucosyltransferase on glucan molecules on the cell surface is not required for the adherence of S. mutans, but de novo glucan synthesis is important in the adherence process.


Use of an alkali-soluble water-soluble extract of Blastomyces dermatitidis yeast-phase cell walls and isoelectrically focused components in peripheral lymphocyte transformations. An alkali-soluble water-soluble extract of Blastomyces dermatitidis yeast-phase cell walls was tested for its ability to elicit a response in lymphocytes isolated from the peripheral blood of Blastomyces-infected guinea pigs. Sequential preparations of the antigen were reproducible and specific in the in vitro lymphocyte transformation assay. Cross-reactivity of the antigen was not evident in lymphocyte transformation assays on lymphocytes obtained from Histoplasma-infected guinea pigs or from animals sensitized with complete Freund adjuvant. Fractionation of the antigen was accomplished on an isoelectric-focusing column, using a sucrose density gradient support. Components were assayed for activity in skin testing and lymphocyte transformation. Comparison of column fractions to the whole antigen showed greater response to the whole antigen in in vivo and in vitro assays.


Genetic control of murine resistance to Toxoplasma gondii. The genetics of murine susceptibility to Toxoplasma gondii was investigated in inbred mice and their F1 and F2 offspring. Among four strains of congenic mice of the B10 background, those with H-2a/a and H-2b/b genotypes were more susceptible than were those with H-2d/d and H-2k/k genotypes. Breeding studies utilizing three of these strains demonstrated linkage between the H-2a allele and greater susceptibility. These data suggest the existence of an H-2-linked gene affecting susceptibility to T. gondii. In challenge of recombinant inbred mice derived from C57Bl/6J (high susceptibility) and BALB/c (low susceptibility) strains, lines BE, BJ, and BK were more susceptible than lines BD, BG, BH, and BI. These data are consistent with the existence of a second disease susceptibility gene linked to the H-13 locus. F1 offspring of the C57B1/6J X B10.D2 mice were significantly less susceptible than either parent. This phenotypic complementary suggests the presence of more than one genetic mechanism of resistance to T. gondii. From these combined data, we conclude that (i) susceptibility to T. gondii in mice is affected by at least two genes, (ii) one of the genes is linked to the H-2 and one to the H-13 locus, and (iii) more than a single mechanism of resistance must be considered to explain the observed genetic controls of susceptibility.


Effect of indomethacin in vivo on humoral and cellular immunity in humans. We studied the effect of indomethacin on intradermal skin testing and antibody responses in humans. Since we and others have shown that prostaglandins are suppressor cell mediators, it was probable that in vivo inhibition of prostaglandin synthesis might enhance the humoral and/or cellular immune response. Administration of indomethacin (Indocin) in a dosage of 100 mg/day to 15 normal men and women resulted in a significantly increased antibody titer to A-Victoria (P less than 0.025) as compared with age- and sex-matched controls. There was no difference in titer to A-New Jersey. Since 90% of the subjects had antibody titers to A-Victoria before inoculation, whereas none had detectable titers to A-New Jersey, we interpret this data as suggesting that indomethacin enhances the secondary but not the primary humoral immune response. Indomethacin administration did not alter the intradermal skin test responses.


Enhancement of experimental anaerobic infections by blood, hemoglobin, and hemostatic agents. Certain foreign materials have been demonstrated to enhance the infectivity of aerobic and anaerobic bacteria. Whole blood and other protein compounds encountered in surgical settings or trauma were tested for their effect on infectivity of nonsporeforming anaerobic bacteria. Infectious synergistic mixtures of Bacteroides fragilis plus Peptostreptococcus anaerobius and Bacteroides melaninogenicus plus Fusobacterium necrophorum were each diluted to a barely noninfectious or minimally infectious concentration (subinfective inoculum) that was injected intraperitoneally into mice alone and in combination with test proteins. Infectivity was measured by deaths from sepsis or abscess(es) within the abdominal cavity at autopsy at 1 week. Two hemostatic agents, Gelfoam powder and Avitene (final concentrations, 10 mg/ml), and crystalline hemoglobin (4 g/100 ml) each produced a marked increase (P < 0.001) in the rate of infection when mixed with a normally subinfective inoculum of either bacterial mixture. Fresh homologous mouse blood (0.25 ml) injected intraperitoneally without anticoagulant also significantly enhanced infectivity (P < 0.01) of a subinfective inoculum of B. fragilis plus P. anaerobius. These studies demonstrated the capacity of whole blood, hemoglobin, and hemostatic agents to enhance the infectivity of certain nonsporeforming anaerobic bacteria. The high concentrations of anaerobic bacteria in the gastrointestinal, female genital, and respiratory tracts produce an increased risk of human infection after surgery or trauma in these sites; the protein agents studied here may further enhance infection.


Streptococcus mutans dextransucrase: stimulation of glucan formation by phosphoglycerides. Lysophosphatidylcholine (LPC) and other phosphoglycerides stimulated water-insoluble and water-soluble glucan production by the Streptococcus mutans 6715 dextransucrase (EC 2.4.1.5). LPC stimulated crude extracellular dextransucrase 1.7-fold, the water-insoluble glucan-producing alpha form of the enzyme 6.5-fold, the water-soluble glucan-producing beta form of the enzyme 2.1-fold, and the cell-associated dextransucrase 2.0-fold. Kinetic studies demonstrated that LPC did not change the K(m) for sucrose of alpha or beta but increased the maximum velocity of the enzymes. The K(m) for LPC of the alpha enzyme was 10(-5) M. LPC from various sources and synthetic preparations of lauroyl-LPC, myristoyl-LPC, and palmitoyl-LPC all stimulated glucan formation. Portions of phosphoglyceride molecules including fatty acids, phosphatidic acid, glycerophosphoric acid, glycerophos-phorylcholine, and choline, when tested individually or in combinations, did not enhance dextransucrase activity. The increased rates of glucan production caused by LPC and primer dextran were additive. Enzyme incubated with LPC before addition of sucrose was stimulated by dextran primer, and, conversely, enzyme treated with dextran was stimulated by addition of LPC with the sucrose substrate. Thus, dextransucrase can be activated by binding of intact phosphoglyceride molecules to a site on the enzyme that is distinct from either the glucosyl donor or glucosyl acceptor (primer) binding sites. Interactions between the S. mutans dextransucrase and amphipathic phosphoglycerides may explain properties of this enzyme which contribute to the cariogenicity of S. mutans.


Virulence of Streptococcus mutans: comparison of the effects of a coupling sugar and sucrose on certain metabolic activities and cariogenicity. A coupling sugar preparation (sucrose-free [CSSF]), which contains a mixture of sugars, oligosaccharides, and oligosaccharides terminated at the reducing end by sucrose, served as a substrate for growth and acid production by Streptococcus mutans 6715. However, CSSF was a poor substrate for cellular aggregation, glucosyltransferase activity, plaque formation, and adherence of cells to glass surfaces. In the presence of sucrose, CSSF inhibited glucosyltransfer activity and adherence of cells. The substitution of CSSF for sucrose in a rat diet significantly reduced caries score. Furthermore, rats fed diets containing sucrose and CSSF had significantly fewer carious lesions than did rats fed a sucrose diet.


Staphylococcal enterotoxin and thermonuclease production during induced bovine mastitis and the clinical reaction of enterotoxin in udders. Enterotoxin A- and C-producing strains of Staphylococcus aureus and partially and extensively purified enterotoxin A were inoculated into the udder quarters of cows. In the course of experimentally induced mastitis caused by the inoculated S. aureus strain, enterotoxin C but not A was detected in the infected udder. Enterotoxin C was observed in mastitic milk samples at very low S. aureus population levels (10(2) to 10(3) colony-forming units per ml). The results suggest that either the synthesis of enterotoxin C is stimulated in vitro or that growth of S. aureus cells in udders was, in fact, higher than the colony-forming unit values indicated. Thermonuclease was shown to be excreted into mastitic milk at a slower rate than was enterotoxin. An inoculation of 1 microgram of enterotoxin A in autogenic milk returned to the udder caused clinical reactions (swelling, palpation sensitivity, and increase in the level of somatic cells) within 6 h.


Mechanisms of hemopoietic and immunological dysfunction induced by lymphocytic choriomeningitis virus. Sublethal irradiation (500 R) of C3H mice is followed by a gradual replacement of radiosensitive cells in their spleens by surviving stem cells originating in bone marrow. This compensatory hemopoiesis was quantitated by counting the numbers of stem cell-derived colonies appearing on spleen surfaces, as well as those which grew in vitro after marrow cells, suspended in soft agar, were overlaid onto syngenic mouse embryo fibroblast feeder layers. Compensatory colony formation, both in vivo and in vitro, was severely depressed when mice were infected with lymphocytic choriomeningitis virus (LCMV) 1 day before irradiation, although the induction of virus-specific cytotoxic T cells in their spleens was unimpaired. Without irradiation, mice, acutely infected with LCMV, showed a dramatic reduction in the numbers of specific antibody-forming cells generated in their spleens after priming with sheep erythrocytes during week 1 post-infection, yet the ability of their marrow cells to form colonies in vitro remained normal. Therefore, the basis of immunodepression is distinct from that of defective hemopoiesis since the latter is apparent only when LCMV infection is accompanied by irradiation. However, as discussed, both phenomena may be related to alterations induced within the splenic environment by LCMV.


Influenza in ferrets and guinea pigs: effect on cell-mediated immunity. Two different animal models were studied to determine whether localized upper respiratory tract viral infection was associated with suppression of systemic cell-mediated immunity. During influenza infection in ferrets, there was no significant decrease in lymphocyte responsiveness to phytohemagglutinin (PHA). Guinea pigs given influenza showed no significant change in their response to PHA or to picryl human serum albumin (picHSA), to which they had been immunized previously. Delayed hypersensitivity skin test responses to picHSA in guinea pigs remained intact during infection. No change in the percentage of circulating T lymphocytes was detected during influenza infection. Transfer of immunity to nonsensitized recipient guinea pigs from picHSA-sensitized guinea pigs was accomplished during influenza infection. Lack of a suppressive effect on systemic cell-mediated immunity after influenza challenge in these two animal models of mild influenza confirmed previous findings in humans with mild influenza infection.


Protein-free and low-protein media for the cultivation of Leptospira. A protein-free medium composed of charcoal-detoxified Tweens (polysorbates), vitamins B12 and B1, inorganic salts, and organic buffer is described that supports the growth and subculture of pathogenic and saprophytic Leptospira. Growth was initiated from small inocula, and cell densities of 10(9) organisms per ml were attained. Antigenicity and immunogenicity of Leptospira cultivated in this medium were similar to those of cells cultivated in serum-containing media. The protein-free medium was converted to a low-protein medium by the addition of 0.1% bovine serum albumin.


Production of antibodies to human interferons in mice. Neutralizing antibodies were raised in mice that had been inoculated repeatedly with moderate quantities of human leukocyte interferon highly purified by affinity chromatography on immobilized anti-interferon globulins. Interferon preparations of lesser purity sensitized the mice to subsequent inoculations of interferon and almost invariably caused death before anti-interferon titers developed. Antibody-purified interferon stabilized by sodium dodecyl sulfate was a superior antigen to interferon that had received mouse serum albumin as an additive. The amount of antibody could be augmented by experimental induction of ascites. The antibodies specifically neutralized leukocyte and lymphoblastoid interferons but not those interferons obtained cultures of human foreskin fibroblasts, embryonic kidney cells, and amnion cells.


Local (immunoglobulin A) immune response by the intestine to cholera toxin and its partial suppression with combined systemic and intra-intestinal immunization. Chronically isolated Thiry-Vella (T-V) ileal loops in rabbits were used to study the local and systemic immune response to purified cholera toxin (CT). Immunization consisted of intraloop (i.l.), subcutaneous (s.c.), or combined i.l. and s.c. inoculation of CT. Fluid from the loops and sera were tested for neutralization of CT by the blueing test and for relative content of isotype-specific (immunoglobulins A [IgA] and G [IgG]) anti-CT. To demonstrate protection against CT, fluid production by the chronic T-V loops was measured after challenge with CT; an "acute" loop prepared from adjacent intestine at the time of challenge was also tested in some animals. The highest neutralizing titers in loop fluids were found in animals receiving i.l. or i.l. and s.c. inoculations, whereas titers in sera were highest in rabbits receiving s.c. or i.l. and s.c. inoculations. IgA anti-CT in fluids became greatest after i.l. inoculation alone and was lowest in s.c. animals. Combined s.c. and i.l. immunization was accompanied by reduced content of IgA anti-CT in fluids as compared with that obtained with i.l. inoculation alone. This finding strongly suggested a suppressive effect on local immunization by s.c. inoculation. While this suppression may have been due to a direct (toxigenic) effect of CT on lymphocytes, an immunogenic mechanism, probably mediated through suppressor T cells, is favored. Little IgG anti-CT was detected in any loop fluids, but high levels were found in sera after two s.c. inoculations or four i.l. inoculations. Neutralization titers for the fluid specimens showed much better correlation with IgA anti-CT values than with IgG anti-CT values. The chronic and acute T-V loops showed protection against fluid production after exposure to CT in systemically and locally immunized animals. However, IgG anti-CT usually appeared in both loops; leakage of serum antibodies because of surgical manipulation was felt, therefore, to invalidate these protection results as a demonstration of local immunity. In challenge studies in undisturbed chronic loops, only local immunization alone was found to result in definite protection.


Kinetics of phagocytosis of Chlamydia psittaci by mouse fibroblasts (L cells): separation of the attachment and ingestion stages. The kinetics of phagocytosis of Chlamydia psittaci (6BC) by monolayers of mouse fibroblasts (L cells) was studied with an assay that distinguished between the attachment and ingestion phases of phagocytosis. At multiplicities of 10 and 100 50% infectious doses (ID50) per L cell, virtually all of the inoculated C. psittaci had been attached and ingested after 60 min at 37 degrees C. At multiplicities of 500 to 5,000 ID50 per L cell, the initial rates of attachment and ingestion of C. psittaci to L cells increased with the multiplicity of infection, but phagocytosis stopped even though many chlamydial cells remained free in suspension and readily available for attachment to the host-cell monolayers. Phagocytosis probably ceased because the L cells were injured when they took up large numbers of chlamydial cells. This injury prevented direct determination of the number of potential binding sites for C. psittaci on each L cell. However, this number is large enough to make the rates of chlamydial attachment and ingestion predominantly dependent on the multiplicity of infection.


Electron microscopic study on phagocytosis of staphylococci by mouse peritoneal macrophages. Macrophages from the mouse peritoneal cavity were examined by electron microscopy at various time intervals up to 24 h after intraperitoneal administration of each of three strains of staphylococci different in virulence for mice: high-virulent, low-virulent, and avirulent strains. After engulfment, avirulent bacteria were highly liable to intracellular digestion, resulting in almost complete degradation within 24 h after injection, whereas high-virulent bacteria were more resistant to digestion, some showing figures suggestive of a dividing process; the gross configuration of most of the ingested bacteria was relatively well preserved over the 24-h period. Time-dependent morphological changes of low-virulent bacteria were intermediate. Among the most distinct cytoplasmic responses to the ingested bacteria was the formation of phagocytic vacuoles around them, the type of which was dependent on the staphylococcal strain infected; ingestion of avirulent bacteria led to formation of vacuoles in which the bacteria were surrounded by a halo of amorphous material of moderate density, which may be the lysosomal content. In contrast, larger vacuoles developed after ingestion of high-virulent bacteria and contained only a small quantity of such amorphous materials. Both types of phagocytic vacuoles were seen around the low-virulent bacteria ingested. Some degenerative changes were found in the macrophages ingesting high- or low-virulent bacteria, but were not in those ingesting avirulent bacteria. Thus, resistance to intracellular degradation, as well as cytotoxicity toward phagocytes of staphylococcal strains, can be correlated with their virulence.


Mode of action of immunological adjuvants: some physicochemical factors influencing the effectivity of polyacrylic adjuvants. The adjuvant effects of different polyacrylic products and monomers were tested. Influenza vaccine was used as a model antigen. Addition of monomers resulted in a decrease in the antibody response, though adjuvant activity of the monomers should be expected according to some theories on adjuvant action. The particle size of the polymer adjuvants proved to be a very important parameter for adjuvant activity. Particles of 0.1 to 0.2 micron yielded a good adjuvant effect, whereas conglomerates or particles bigger than 0.5 micron yielded only poor or no adjuvant effects. The adjuvant effect of 0.1- to 0.2-micron particles was much more reproducible than rat of Al(OH)3. Attention is drawn to the importance of using physiochemically reproducible materials, such as polymer particles, for experimental work.


Immunoglobulins and secretory component in the external secretions of foals with combined immunodeficiency. Nasal washings and tears were collected from seven Arabian foals with combined immunodeficiency and nine normal foals. The major immunoglobulin in the external secretions of normal foals over 2 months of age was secretory immunoglobulin A, whereas foals with combined immunodeficiency lacked this immunoglobulin. The external secretions of both normal and immunodeficient foals contained free secretory component at birth.


Aging in vitro and large-scale interferon production by 15 new strains of human diploid fibroblasts. To develop resources for large-scale production of human fibroblast interferon, we isolated, cryopreserved, and characterized 15 new strains of human diploid foreskin fibroblasts. Their life spans in vitro ranged from 52 to 72 population doublings. We based the selection of cell strains for mass interferon production on the number of population doublings during which consistently high yields of interferon were obtained after "superinduction" in roller bottles; our data show that aging in vitro leads to significant decline in amounts of interferon produced. In contrast, susceptibility to interferon remains largely unaffected by in vitro senescence. Karyotypic analysis indicated that the best interferon-producing strain, MLD (over 60,000 reference units/ml), has a translocation between chromosomes 5 and 15.


Detection of cholera enterotoxin activity in suckling hamsters. For detection of cholera enterotoxin activity, suckling hamster, mouse, and rat models were compared. It was found that not only suckling mice but also suckling rats and hamsters were sensitive to cholera enterotoxin. Among these models, suckling hamsters were the most sensitive and gave positive results with about 1/100 the cholera toxin seen in suckling mice and rats.


Amphotericin B concentrations in saliva after application of 2% amphotericin B in orabase. 2% amphotericin B in orabase was applied to the buccal mucous membranes of healthy volunteers. Amounts of 1 g and 0.5 g were used. The experiments were performed during the day time as well as during the sleeping hours at night. At several intervals after application saliva samples were collected and the amphotericin B concentrations were determined. Application of 1 g of orabase with 2% amphotericin B appears to maintain a sufficiently high concentration for four hours, 0.5 g for three hours. Application of these amounts before going to sleep guarantees a sufficient concentration until awakening the next morning.


The prevalence of ay and ad subtypes of the hepatitis B surface antigen in Belgium. Subtyping in various groups of persons positive for hepatitis B surface antigen gave the following results: Among asymptomatic blood donors (healthy carriers) subtype ad was found in 20, subtype ay in 24. Among symptomatic patients with acute hepatitis B ad was found in 25, ay in 29. Among patients of an institution for the mentally retarded ad was found in 9, ay in 23. Among patients of a Among patients of a geriatric institution ay was found in 9; among medical personnel of the same geriatric institution ay was found in 3 persons. Among symphilis patients ad was found in 6, ay in 29; and in patients of a dialysis centre ay was found in 6. These data show that neither subtype is really dominant in Belgium.


Assays of cephalosporin antibiotics administered prophylactically in open heart surgery. Determination of serum and tissue levels before, during and after cardiopulmonary bypass. Twenty-eight patients who underwent open-heart surgery were divided into three groups, each of which received a different antibiotic from the cephalosporin series (cephalotin, cefazolin or cefamandole) in order to prevent infection. All antibiotics were given via intravenous infusion in a dosage of 2 g prior to surgery. To clarify the question of antibacterial activity under operative conditions with the cardiopulmonary bypass, the serum and tissue levels were determined before, during and after the surgical procedure. The effectiveness of the cephalosporins against bacteria most frequently encountered in open-heart surgery was demonstrated and substantiated by the serum and tissue concentrations. It became apparent that, in view of the favorable serum and tissue levels during and after the cardiopulmonary bypass, cefamandole should be considered the antibiotic of choice in preventing infections during open-heart surgery.


Polymyositis accompanying coxsackie virus B2 infection. A case of polymyositis in a 15 year old girl demonstrating the clinical course of an infectious disease is presented. The Coxsackie virus B2 was isolated from the stools, and serum antibodies were detected. Histologic examination of muscle showed signs of myositis. The patient was treated with cortisone and recovered completely. Possible viral etiology of the polymyositis is suggested.


Leucocyte migration inhibition with Armadillo lepromin in human lymphocytes of healthy donors. Armadillo lepromin activates human lymphocytes from normal donors to release leucocyte inhibiting factor. The above activity was expressed optimally when leucocytes were incubated with lepromin at 37 degrees C, and only partially when incubation was carried out at 30 degrees C or at 35 degrees C. The possible mechanism of the in vitro production of lymphokine from lymphocytes stimulated by armadillo lepromin is discussed.


Ventriculitis, aqueductal stenosis and hydrocephalus in neonatal meningitis: diagnose and treatment. Five newborns are presented who had meningitis and developed aqueductal stenosis. The diagnosis of obstruction was made by ventriculography during the course of meningitis. A variety of therapeutic measures (use of intraventricular antibiotics, use of reliably diffusible antibiotics, drainage of ventricles) led to varying results. Protocols should be established to assess each modality or combination thereof to help reduce the extremely high morbidity and mortality of this disease.


Characterization of surface receptors on bovine leukocytes. Three bovine leukocyte populations--peripheral blood lymphocytes (PBL), mammary gland polymorphonuclear neutrophils (PMN) and macrophages (Mo)--were characterized with respect to five surface markers: surface immunoglobulin (SIg), sheep erythrocyte receptor, complement (C) receptor and Fc receptors for both IgG and IgM. The majority of PMN and Mo possessed C and Fc receptors for IgG, but lacked SIg and the erythrocyte receptor. The PMN, but not Mo, also expressed a Fc receptor for IgM. The PBL were heterogeneous with respect to their surface characteristics and evidence was presented for the following subtypes: (a) cells with the E receptor alone; (b) cells with E receptor plus the Fc(IgG) receptor; (c) cells with SIg plus the C receptor but minus the Fc(IgG) receptor; (d) lymphocytes with SIg plus the C receptor and the Fc(IgG) receptor, and (e) cells lacking E receptors and SIg but bearing Fc(IgG). It was assumed, but not proven, that some of these latter cells must also bear the C receptor. The significance of the various cell types in antiviral defense is briefly discussed.


Glucans and dextrans in rats and man.

Glucans and dextrans in rats and man. Intracutaneous injections of three glucan contaminants of invert sugar solutions and crude cane sugar into human skin produced localised wheals and erythema reactions. These glucans of molecular weight between 12,000 and 1,400,000 were also active on intradermal injection into both dextran-sensitive and dextran-resistant rats, and, like dextrans, were active on subcutaneous injection into dextran-sensitive animals.


Studies of the paw test and tumor-associated immunosuppression in mice. Specific paw test reactivity to tumor cells was transferred to normal mice using lymphoid cells or serum from mice with a growing or surgically excised methylcholanthrene-induced transplantable sarcoma. When the donors had been sensitized to PPD and to tumor, normal recipients of lymphoid cells became reactive by specific paw test to tumor cells and PPD. Normal recipients of serum from the same donors also became reactive to tumor cells. Normal recipients of serum from tumor-bearing mice developed depressed PHA paw tests. Adoptive transfer with cells or serum of paw test reactivity to tumor and PPD was unsuccessful when recipients had large tumors. This was the case when the recipients had either tumor of the same line or that of a separate line that did not cross-react antigenically.


Properties of receptors for IgG on human placental cell membranes. The amount of 125I-IgG which bound to membranes isolated from the human placenta was competitively inhibited by the presence of increasing amounts of unlabeled IgG but not by unlabeled albumin. The relationship between membrane-bound and free IgG indicated the presence of membrane receptors with an appreciable affinity for IgG. Incubation of membranes with collagenase or neuraminidase did not results in appreciable reduction of IgG-membrane binding, indicating that neither intact collagen nor sialic acid  play an important role in the binding. Placental surface membranes isolated by salt extraction bound 3.79+/- 1.78 (SD) pmol IgG/microgram membrane protein, whereas membranes isolated by differential centrifugation bound only 1.61 +/- 0.24 pmol/microgram (p less than 0.02). The fraction of a preparation of solubilized membranes which bound to an IgG affinity column yielded on polyacrylamide gel electrophoresis three prominent protein bands which had molecular weights of 3.7 X 10(4), 4.5 X 10(4) and 6.0 X 10(4) daltons. These findings are consistent with the existence of a limited number of receptors for IgG on placental membranes, including IgG receptors on the microvillus membrane of the syncytial trophoblast. The latter, in accordance with Brambell's hypothesis, could be of importance in the transplacental transport of maternal IgG.


Studies on the role of histamine and 5-hydroxytryptamine in immunity against the nematode Trichostrongylus colubriformis. IV. Inhibition of the expulsion of worms transplanted into the duodenum of immune guinea pigs. Significant numbers of fourth larval stage Trichostrongylus colubriformis from sheep donors are expelled from the small intestine of T. colubriformis-immune guinea pigs within 8 h of transplantation into this organ. This model system for the immune expulsion of gastrointestinal nematode parasites was used in the current experiments to examine the effect of drugs recently shown to modify the release of histamine. The experiments showed that worm expulsion was inhibited by the beta-adrenergic agonists salbutamol and isoprenaline, the phosphodiesterase inhibitors aminophylline and ICI 63,197, the antiallergic drug ICI 74,917 and the antihistamine mepyramine. The beta-blocker propranolol prevented inhibition of worm expulsion following salbutamol treatment and there was some evidence of synergistic action between salbutamol and ICI 63,197. The results support previous findings suggesting an important role for histamine release in the effector mechanism of the immune response of guinea pigs against the parasitic nematode T.colubriformis.


Quantitation of IgE on the basophil surface in atopic patient. A radioimmunoassay was employed in order to quantitate IgE on the basophil surface in 6 normal and 10 pollen sensitive subjects. A basophil-rich fraction obtained from whole blood in a Ficoll gradient was incubated with specific anti-IgE; the inhibition produced by surface IgE was standard inhibition curve. Atopic patients had on the average about 10-30 times as many IgE molecules per basophil as control subjects. Total and specific IgE as well as degranulation by Lollium perenne crude extract were also studied in both populations and their results compared with those of basophil IgE quantitation.


Human colostral macrophages as indicator cells in migration inhibition test. The macrophage inhibition test was carried out using colostral macrophages obtained from PPD sensitive and measles and mycoplasma pneumoniae antigen insensitive woman. The macrophage migration was inhibited by PPD but neither by measles nor by mycoplasma pneumoniae antigen. Human colostral macrophages can be used as indicator cells in migration inhibition tests.


Immunoglobulin class distribution of measles virus antibodies in serum and spinal fluids of patients with subacute sclerosing panencephalitis. Sera and cerebrospinal fluids (CSF) from patients with subacute sclerosing panencephalitis (SSPE) were tested by indirect immunofluorescence for the presence of measles virus specific antibodies of the various heavy chain classes. IgG and IgA antibodies were detected in the CSF while IgG, IgM, IgA, IgD and IgE measles virus antibodies were found in a significant number of the patient sera. Sera from SSPE patients had slightly elevated levels of IgG, IgM, IgD and IgE while IgA was decreased. The heterogeneous heavy chain class distribution of measles antibodies suggests the possibility that non-complement fixing antibodies serve as blocking antibodies which aid in the persistence of intracellular measles virus infection in patients with SSPE.


Prostaglandins and inflammatory cell movement in vitro. The prostaglandins PGE1, E2, F2alpha, and A1, had no significant cell migration inhibitory or chemotactic activity for guinea pig peritoneal exudate cells when tested in vitro using respectively the capillary tube cell migration test and the Boyden chamber test for chemotaxis. PGE1 and PGE2 also did not affect the positive chemotactic action of lymphokine. The proposed role of prostaglandins as chemotactic agents in inflammatory responses therefore should be treated with some caution.


Studies of contact hypersensitivity and tolerance in vivo and in vitro. I. Basic characteristics of the reactions and confirmation of an immune response in tolerant mice. Contact hypersensitivity to dinitrochlorobenzene (DNCB), picryl chloride (PCl) or oxazolone was induced in mice by skin painting with these agents, and was measured by skin testing in vivo using the ear swelling method. Prior administration of dinitrobenzene sulfonate (DNBS) or picryl sulfonic acid prevented sensitization to DNCB and PCl, respectively. Both this tolerization and the original sensitization were specific for the hapten used. Cyclophosphamide given before sensitization enhanced skin reactions, but when given before tolerization it interfered with establishment of tolerance. Cells from both sensitized and tolerized mice were shown to be reactive with the corresponding haptens in vitro in the leukocyte adherence inhibition (LAI) reaction. LAI specificity was similar to that found for cutaneous reactivity. The reaction of DNCB-sensitized cells with DNBS led to the production of a soluble mediator which induced LAI in normal cells. The demonstration of potentially reactive cells in mice judged to be tolerant by skin testing indicates the concomitant existence of suppressor factors.


Modulation of eosinophil chemotaxis and eosinophil chemotactic factor release by concanavalin A. Concanavalin A (Con A) is not a chemotactic factor for guinea pig eosinophil leukocytes. It does, however, modulate the response of cells to neutrophil-derived eosinophil chemotactic factor (ECF) or to C5a. This effect occurs by a direct interaction of Con A with the target cell and is reversible by appropriate doses of alpha-methyl-D-mannoside (alpha-MM). The inhibition of chemotaxis at high doses of Con A is due to agglutination of the cells on top of the filter. The mechanism of enhancement at low doses is not understood, but several possible explanations are explored. Careful definition of the effect of Con A on eosinophil chemotaxis makes it possible to exclude an effect of Con A on ECF secretion by neutrophils.


Levels of homocytotropic antibody in hereditarily asplenic, splenectomized and normal mice. No significant differences were detected in circulating levels of IgE or IgG1 in hereditarily asplenic (Dh/+) or splenectomized mice compared to normal controls (+/+). These findings suggest that the spleen plays a minor role in the regulation of IgG1 and IgE production in this mouse line.


Detection of trophoblastic tumour activity by pregnancy-specific beta-1-glycoprotein. Pregnancy-specific beta-1-glycoprotein (PSBG) and human chorionic gonagotrophin (hCG) were measured by specific radioimmunoassays in 73 serum samples from eight patients treated for gestational choriocarcinoma. Both markers were initially present in all cases, but at the time of writing both had levelled off in six cases and decreased markedly in two. Usually hCG was detectable for longer than PSBG during the course of chemotherapy. Both PSBG and hCG reappeared on clinical recurrence. However, at times during the follow-up period one trophoblastic marker only was expressed. This was hCG in most cases, but in one case PSBG alone was encountered. These results suggest that PSBG may serve as an additional marker for the detection of residual tumour growth in patients with trophoblastic tumours.


Movement of human mammary tumour cells in culture: exclusion of fibroblasts by epithelial territories. Time-lapse cinematography was used to investigate the movement of confronting populations of human mammary epithelium and stromal cells (fibroblasts). Epithelial cell islands from fibroadenomas and from normal lacteal secretions completely excluded the fibroblasts, and individual cell territories were maintained even in dense cultures. Electron microscopy of the boundary between epithelium and fibroblasts showed that the two cell types made contact. In contrast, epithelial islands from two carcinomas did not retain territorial integrity and allowed penetration of mammary fibroblasts. Confronting homologous eptihelial islands from benign tumours merged, but this was shown to be due to interdigitation rather than free mixing of cell. Epithelial cells moved actively but unlike fibroblasts they retained their neighbour relationships.


Isolation of a human prostate carcinoma cell line (DU 145). A long-term tissue culture cell line has been derived from a human prostate adenocarcinoma metastatic to the brain. The cell line, DU 145, has been passaged 90 times in vitro over a period of 2 years. The cells are epithelial, grow in isolated islands on plastic Petri dishes, and form colonies in soft agar suspension culture. Karyotypic analysis demonstrates an aneuploid human karyotype with a modal chromosome number of 64. Distinctive marker chromosomes (a translocation Y chromosome, metacentric minute chromosomes and three large acrocentic chromosomes) have been identified. Electron microscopy of the original tumor tissue and of the tissue culture cell line show a remarkable similarity in cell organelle structure.


Mobility of cells from solid tumors.

Mobility of cells from solid tumors. The mobility of cells from two established cell lines of hamster origin, NIL B and its SV40 virus-transformant SV-NIL, was studied in cellular aggregates maintained in agitated liquid medium. Subcutaneous injection of 5 x 10(6) NIL B or SV-NIL cells into Syrian hamsters resulted in a high frequency of tumor formation - 1.00 for SV-NIL cells and 0.93 for NIL B cells. Cells from five different tumors of NIL B origin were grown in tissue culture and their mobility in cellular aggregates was also studied. The mobilities of the tumor-derived cells were similar to each other and to that of the SV-NIL cells, but slightly higher than that of the NIL B cells. In addition, the plating efficiency, saturation density and doubling time of NIL B, SV-NIL and the tumor-derived cells were determined in conventional flat culture. No consistent pattern of saturation density or doubling time was observed in the tumor-derived cells with respect to each other or to the established lines; however, the plating efficiencies of the tumor-derived cells were all considerably lower than those of NIL B and SV-NIL cells. It was concluded that selection for the ability to divide and survive in vivo (i.e. to form tumors) was accompanied by a modest increase in cell mobility in vitro.


Separation of lymphoid cells with a suppressor effect on the activity of cytotoxic cells in vitro during the growth of a syngeneic mouse tumour. One or 4 weeks after grafting of a syngeneic sarcoma (T2) to C57BL/6 mice, lymph-node cells (LNC) are cytotoxic in vitro for the cells of this tumour. But after 2 weeks LNC are not cytotoxic or show a non-significant activity. These second-week LNC, added to cytotoxic lymphocytes (CL) from the first or fourth week, reduce considerably the cytotoxicity of the latter cells. When velocity sedimentation at unit gravity is used to fractionate 11th-14th day LNC, some fractions, enriched in small lympoid cells, kill the cancer cells. Other fractions, containing large blast cells, lack this property but can suppress the activity of the small cells or of fourth week CL. These suppressor cells can also be separated from active CL by passage on a glass bead column. They are "adherent" while CL are non-adherent at this stage. This suppressor effect is abolished when the lymphoid cell suspension is treated by anti-theta serum, but macrophage depletion does not modify the inhibition. The cell responsible for the suppressor effect is thus a large T lymphoid cell, adherent and non-phagocytic. It seems to act essentially on an effector phase of the cell-mediated cytotoxicity.


Growth characteristics in vitro of hybrids between normal and transformed cell lines. Sensitivity to the inhibition of division during cell crowding in vitro was determined for hybrids between normal and tumor cell populations of human and mouse origin. Interspecies hybrids were more sensitive to cell crowding than intraspecies hybrids. The results suggest that the inhibition of cell division due to crowding was more dependent upon the species of origin rather than on the normal or transformed character of the parental cell lines of the hybrid cell populations.


Cell surface receptors for ecotropic MuLV: detection and tissue distributions of free receptors in vivo. The expression of available cell surface receptors for binding of the major envelope glycoprotein of Rauscher MuLV, gp71, was studied in freshly prepared tissues of the low-leukemic mouse strain CBA. The presence of such receptors is a requirement for infection and therefore also for transformation of cells in vitro by ecotropic MuLV. This paper describes the conditions of the binding assay for radiolabelled RLV gp71 and freshly prepared lymphoid cells under which binding occurs as a function of time and cell number. With this assay, available receptors for RLV gp71 were detected in all lymphoid as well as some non-lymphoid tissues, most notably brain. The lack of binding in other non-lymphoid tissues was not due to the presence of gp71 specified by endogenous ecotropic MuLV, as was determined by a sensitive group specific RIA. These results demonstrate that the distribution of receptor sites necessary for infection by ecotropic MuLV does not directly correlate with the tissue distribution of malignant transformation that is most commonly induced by these endogenous viruses. Therefore, susceptibility to infection by ecotropic MuLV, as determined by the expression of viral receptors, is not sufficient to determine the target organ for MuLV-induced oncogenesis.


Polyadenylated RNA molecules and polyribosomes in tumours of chemical and viral origin. The distribution of poly(A)-enriched mRNA in nuclei, mitochondira, free and membrane-bound polyribosomes from normal C3HA mouse and Syrian hamster livers and normal chicken fibroblasts has been compared with that in corresponding subcellular fractions in a transplantable, chemically induced MD hepatoma, non-virus-producer hamster and virus-producer chicken Rous sarcomas. It has been shown that the content of poly(A)-RNA is increased in all tumour fractions except in free polyribosomes. The distribution of different classes of polysomes i.e. free, membrane-bound and mitochondrial outer-membrane-associated polysomes in tumour cells was changed in comparison to that in normal cells. It is concluded that in tumours of chemical and viral origin, the observed changes in the two components of the protein-synthesizing apparatus occur simultaneously.


In vitro and in vivo indications of the carcinogenicity and toxicity of food dyes. Eight food dyes or commercial color mixtures certified for use in the United States were tested for their ability to transform in vitro a serial line of Fischer rat embryo cells previously reported to be a sensitive indicator of chemicals having carcinogenic potential. Malignant cell transformation was induced by a commercial mixture (G2024) of two of these dyes (Blue 1 and Yellow 5) and by Blue 2, Green 3 (one of two experiments) and Red 4. Food dyes Blue 1, Red 3, Yellow 5 and Yellow 6 did not induce cell transformation. One to 1.5 mg of each dye was injected into suckling LVG or Graffi hamsters which were monitored for tumor induction and/or death over a 330-day period. None of the non-transforming dyes (Blue 1, Red 3, Yellow 5, Yellow 6) or Green 3 induced a significant increase in tumor (mostly lymphoma) incidence or animal mortality. Three of the transforming dyes (Blue 2, Green 2024, Red 4) did increase tumor incidence and/or mortality in at least one strain of hamster. We conclude the the in vitro assay suggested that certain food dyes were carcinogens and that in vivo studies in hamsters supported this interpretation.


Isoniazid tumorigenicity in mice under different experimental conditions. Tumorigenicity of isoniazid in Swiss and A strain mice was studied with three different doses (0.55, 1.1 and 2.2 mg/mouse/day). Mice were fed isoniazid by gastric intubation and killed when they appeared weak. Incidence of tumour ranged from 32 to 100%. Subsequently sex-specificity in isoniazid tumorigenicity was also studied as well as the effect of vitamin B complex or protein deficiency, and of transplacental exposure of Swiss strain embryos to isoniazid. It was observed that protein deficiency and transplacental exposure stimulated tumour incidence in mice treated with 0.55 mg of isoniazid. No sex specificity was observed in isoniazid tumorigenesis.