Panic-fear in asthma: requests for as-needed medications in relation to pulmonary function measurements. Requests for as-needed medications and treatments (PRNs) by asthmatic patients scoring high, moderate, or low on the Asthma Symptom Checklist panic-fear category were studied for days when patients were matched at normal, intermediate, and subnormal levels of pulmonary function. Low panic-fear patients were the least likely to request PRNs regardless of the pulmonary function level. In contrast, high panic-fear patients often requested PRNs each level of pulmonary function. Only moderate panic-fear patients made progressively more PRN requests on days when pulmonary functions were lower. These observations and others concerning the adverse influence of extreme panic-feat coping styles upon the treatment of asthma were discussed.


What should be optometry's role? How should we prepare for it? This is a position paper which broadly states the role which, in the author's opinion, optometry should adopt as its goal and the educational environment in which we can best prepare graduates to fulfill that role. The role briefly stated, is that of first contact supplier of vision care and of certain aspects of primary health care. The appropriate educational milieu is held to be the academic health center.


Optometric goals and priorities.

Optometric goals and priorities. An analysis of the needs of the public for optometric services by (1) the examination of the epidemiology of vision problems, (2) a systems analysis of the vision care system, (3) the services optometrists provide and (4) the educational resources available, indicate that the priorities for optometric development should be on primary vision care services and new academic facilities.


Leadership and change in optometric education. The quality of institutional administration and the competence of its academic leaders are considered to be crucial elements in the continued progress of optometric education. Educational leadership to meet the need for ongoing, guided, systematic educational planning is underscored. Such planning, in turn, depends significantly on the emerging role definition of the optometrist as a primary care practitioner.


Financing of optometric education.

Financing of optometric education. Decline in enrollments in college age group is reflected in health professional and law schools. This evolving reduction in applicant pool is discussed with reference to caused factors and contrasted with the post World War II boom. These factors are discussed in relationship to present and future fiscal integrity of our optometric colleges. The author illustrates the need for securing financial support from state governments for existing optometric educational institutions before building new schools.


Education for the optometric educator.

Education for the optometric educator. Optometric education is faced with the need for new thinking in developing and training educators and new methods of financing their education. Establishing of research faculty and the instituting of research in present schools of optometry must be given a high priority in the future of optometric education.


Methodology for interfacing results of basic science research and clinical practice. The importance of the integration of basic vision sciences and applied clinical research is discussed. Steps which must be taken in optometry and in the schools and colleges in particular are reviewed. Some of the problems that will be encountered are discussed.


Quality is still the backbone of good education. The author believes that quality optometric education results from excellent teaching, which depends on qualified faculty members who also add to optometric knowledge through research, and on a curriculum that stresses fundamentals. He believes that practicing optometrists can help to encourage and assist research.


Continuing optometric education crossroads. This paper gives a brief overview of continuing optometric education from the perspective of the providers and the Commission on Continuing Education. The early history is covered as well as a discussion of some present problems and concerns of many of the providers. This is followed by recommendations for the future and a proposed plan for each provider's involvement in continuing education.


The paraoptometric puzzle.

The paraoptometric puzzle. In the past ten years, interest in the production and utilization of paraoptometric personnel has risen sharply. Production alone will not meet the demands of the foreseeable future. More effective utilization of the skills of the trained paraoptometric is vital. Current practitioners have been slow to accept new modes of vision care delivery. Conversely, most optometric educational institutions have been unwilling or unable to develop training programs for student optometrists beyond the traditional solo concept. Recommendations are presented for immediate and long range remediation.


Prevention and enhancement: future needs. The objective of prevention of eye and visual problems, and the objective of enhancement of vision performance are primary and most urgent for undergraduate, postgraduate and/or continuing optometric education. Prevention is something done before the need occurs. The concept of enhancement assumes performance far beyond so-called normals. The future needs of the public and the profession should determine the content and scope of optometric education pre and post graduation. Occupying the unique areas of prevention and enhancement can expand optometry's role, insure its continued growth and vital service to the public.


The optometric recognition award program--an option for self assessment. This discussion addresses itself to the need for all optometrists to continue recognizing their professional responsibility to maintain continuing competency through meaningful optometric education experiences; and to work toward this goal by participation in the Optometric Recognition Award Program. The ORA Program is structured to enable the participant to design, record and fulfill his personal continuing optometric education needs. Fullfillment of the award requirements, 150 hours of C.O.E. in three years, results in recognition by the AOA in the form of a recognition plaque.


Age-dependent changes in the oxidative metabolism in rat brain. The effects of aging on the oxidation of labeled glucose and 3-hydroxybutyrate and on several mitochondrial enzymes in rat brain were investigated. The oxidation of labeled glucose and labeled 3-hydroxybutyrate was diminished by about 40 and 35%, respectively, in cerebral cortex slices from 2-year-old rats compared to those from 3-mo-old animals. A significant reduction in the activities of 3-hydroxybutyrate dehydrogenase, 3-oxo acid CoA transferase, acetoacetyl CoA thiolase, and NAD-isocitrate dehydrogenase was observed in brains of 1- and 2-year-old rats compared to 3-mo-old animals. However, aging had no effect on the activities of citrate synthase and pyruvate carboxylase. These findings show that specific alterations occur in the activities of several mitochondrial enzymes in aging brain.


Quantitative and morphological changes in neurons and neuroglia in the indusium griseum of aging mice. The indusium griseum was investigated quantitatively in 5-mo-old and 18-mo-old mice. There was no difference in the number of neurons between 5 and 18 mo, but the number of neuroglia increased by almost 30%. Astrocytes and microglia increased in number, but oligodendrocyte number remained unchanged. There was no significant variation in the volume density of neuronal perikaryon, neuropil, or blood vessels with age.


Isolated aortic valve stenosis in the eighth decade. Aortic valve stenosis was evaluated in 26 patients in the eighth decade of life, representing 20% of all patients referred for aortic valve stenosis. There were no clinical features which distinguished this older group from younger patients. Coronary artery disease was present in 46% of the older group, 30% in the younger. Angina pectoris or an infarction pattern on the electrocardiogram was not useful in predicting coronary artery disease in either group. A dominant left coronary circulation was more frequent (22%) in the younger than the older group. This was related to the higher incidence of congenitally deformed aortic valves (55%) found in the younger, compared in the advanced age group (5%). The surgical mortality in the younger and advanced age group was 8 and 20%, respectively. Mortality in both groups was related in more than half the patients to coronary artery disease.


A factor analytic study of daydreaming, imaginal process, and temperament: a replication on an adult male life-span sample. One hundred and seventy males aged 24 to 91 years were measured on daydreaming and related mental activity using the imaginal processes inventory and on temperament using the Guilford-Zimmerman Temperament Survey to determine if earlier results relating temperament to daydreaming in college populations were generalizable to the adult life-span. The earlier derived factor of "Neurotic-Anxious Absorption in Daydreaming" was found in this sample and had a negative correlation with chronological age, suggesting a decrease with increasing age. Other earlier derived factors on "Personal Acceptance-Utilization of Daydreaming," "Masculinity-Femininity," "Thinking Introversion," and "Social Extroversion" were also found in this sample, but were not related to age.


Changes in individual attitudes toward traditional morality: a 24-year follow-up study. Changes in individual attitudes toward traditional morality are charted by focusing on a sample of 1950 persons from rural Pennsylvania surveyed as high school sophomores in 1947 and again 24 years later when they were in their early 40s. A series of 12 Likert-type items were used to assess traditionalism. In general, the subjects' attitudes changes toward a more permissive stance in regard to questions concerned with adult behavior. Liberalization of attitudes toward the behavior of young people was less pronounced. The direction and degree of change differed markedly among individuals. Frequency of church attendance was closely associated with the amount of change. Those attending church once a week or more were decidedly less likely to change toward a liberal or permissive position than were those who attended less often. Other factors related to amount of liberalization included residence, education, and income.


Sampling in social gerontology: a method of locating specialized populations. This paper describes a two-stage sampling design for obtaining probability samples of the elderly and other specialized populations. The first stage enumerates a sample of elderly individuals residing in a probability, sample of households; the second stage involves the mailing of a questionnaire to the eligible respondents located in the first stage. The results of this method for a study of noninstitutionalized elderly in Washington State are reported. These results indicate that (1) elderly individuals will respond in the two-stage sampling design does not appear to increase sample bias above that expected in normal mailed-questionnaire studies, and (3) the method is extremely inexpensive. Use of the two-stage design by social gerontologists is recommended.


[Study of folic acid blood levels during estro-progestagen treatments]. When the blood levels of folates were estimated in 100 women following treatment with oestro-progestagens the following facts emerged : 1) There is no significant difference between the folate levels found in women on oestro-progestagens compared with a control population of the same age. 2) We have not found any evidence of frank folic acid deficiency in women on oestro-progestagens, but there have been slight drops in the levels of folates in women who show other reasons for folic acid deficiency than taking oral contraceptives. These are : troubles in absorption, salmonella, an increase in the need for folic acid in pregnancy, and failure of transport. Furthermore the incidence of lower levels in the control population was exactly the same as in the experimental population.


[Dynamic study of tissue circulation in the cervix uteri and the prognosis of the newborn infant]. A new machine has been developed to measure heat dissipated in the tissues especially by convection linked to the blood flow. Its use is simple and without any danger as far as application to the pregnant cervix goes. New discoveries are presented. A new concept of the micro-circulation is considered together with the consequences. The practitioner may be able to draw conclusions as to the prognosis, as to whether a small-for-dates fetus together with or without premature labour is likely. This depends on the tracings obtained which register the intrinsic activity of the periarteriolar muscles.


[Pneumoperitoneum of genital origin. Apropos of a case at the beginning of pregnancy]. The authors studied the literature for similar findings in order to try to classify the different types of pneumoperitoneum when they had a case of pneumoperitoneum of genital origin. They then went on to study the mechanism by which these rare cases of genital pneumoperitoneum occurred: the insufflation or aspiration of air through the tubes, tubal antiperistalsis and uterine or vaginal perforations. From the clinical angle it is essential to eliminate pneumoperitoneum secondary to perforation of a hollow organ, and then to look for signs of peritonitis which only occur in genital pneumoperitoneum due to uterine perforation. The case usually cures spontaneously but recurrences are not uncommon. If there are no signs of peritonitis expectant treatment should usually be undertaken.


[Viral hepatitis and pregnancy].

[Viral hepatitis and pregnancy]. When nine cases of viral hepatitis had been analysed they showed that the normally held pessimism of this infectious pathological condition associated with pregnancy was ill-founded. The natural outcome for viral hepatitis in our country is not any worse in pregnancy than when the patients are not pregnant. The outlook for the fetus is above all determined by its prematurity, since we have not observed any cases of abortion, malformations or small-for-dates fetuses. On the other hand the neo-natal progress is slow with anaemia and jaundice frequent signs. Treatment of threatened early labour with beta-mimetic drug has been carried out carefully because of the risks for the fetuses.


[Treatment of amenorrhea-galactorrhea with bromo-ergocryptine. Apropos of 12 cases]. The authors report the action of bromergocryptine in 12 patients who were suffering from a syndrome of amenorrhoea with galactorrhoea and hyperprolactinism. None of these 12 patients had a pituitary tumour; or else they had been operated on before treatment. Only in one case was the syndrome linked to taking oral contraceptives, which had been stopped more than six months previously. The dose of bromergocryptine was 5 mg. a day. If this dose fails it is possible to raise it after two months' observation. The galactorrhoea disappeared or markedly decreased in 8 out of the 12 cases. Ovulatory cycles started in 9 out of the 12. 5 of the women wanted a pregnancy: 4 succeeded and 2 of them have already been delivered of normal children. Bromergocryptine (CB 154) has thus proved itself to be remarkably effective in hyperprolactinism, not caused by a pituitary tumour, which gives rise to amenorrhoea and galactorrhoea. All the same, its long-term use, especially after stopping treatment, is still to be evaluated.


[The outlook for gross hydrosalpinx].

[The outlook for gross hydrosalpinx]. Classically, the volume of a hydrosalpinx is taken as an essential factor in working out the prognosis of tubal operations carried out on this kind of lesion. This theory is contradicted following a study of a series of 62 patients who presented with greatly dilated ampullae. Firstly, the percentage of cases that were and could not be operated on during examination was no different in this group than in those where the hydrosalpinx was of a more usual volume. Secondly, at least 12 patients, which means 28 per cent of those who had tubal plastic surgery, had at least one intrauterine pregnancy. The percentage of those with tubal patency if also very satisfactory because it could be restored in more than three-quarters of the cases. Remarkably, in this series there was no ectopic pregnancy. It seems, therefore, that it is time to review the poor reputation of the large hydrosalpinx.


[Perinatal mortality. Statistics from the Gynecological and Obstetrical Service of the Notre-Dame-de-Bon-Secours Hospital (1961-1975)]. The authors present their statistics for perinatal mortality from 1961 to 1975. In the last 5 years this mortality is less than 15 per 1000, in spite of an increase in the numbers of pathological pregnancies. Over and above progress in obstetrics and neonatology that has occurred, this result can be attributed in part to the quality of supervision carried out by the senior obstetricians and in part to the existence of pathological consultation which takes place between colleagues about the therapy to be carried out in multidisciplinary meetings.


[Correlation between initial sperm quality and its ability to tolerate cryopreservation]. The aim of this study based on the investigation of 570 semen specimens providing from 251 patients is to determine the correlation existing between initial sperm quality and its ability to bear cryo-preservation. Semen quality was estimated according to the usual parameters : numeration, motility and morphology of spermatozoa. Motility recovery on the one hand, and fertilizing capacity after cryo-preservation on the other hand, allowed an estimation of the sperm ability to bear thawing. Motility recovery seems to depend widely on the initial sperm quality. The persistence of sperm fertilizing capacity partly depends on the motility recovery and consequently partly on the initial quality. However, some semen specimens seem to lose their fertilizing capacity in spite of a good motility after deep-freezing.


[Chronic suppuration of the breast].

[Chronic suppuration of the breast]. We have found a series of 14 cases of chronic suppuration of the breast in 10 years in the Surgical and Gynaecological Clinic of the Salpêtrière Hospital. The symptomatology at the outset is usually innocuous, showing itself as a tumour that appears either gradually or suddenly. Diagnosis is not difficult and is usually solved by tapping the tumour. The progress of these lesions to become chronic with the formation of fistulae is the main risk, and the incident is not negligible. Thus the true problem is to find the correct therapy so as to avoid this possible unfavourable outcome.


[Transplancental exchange of oxygen in the female]. Extra-uterine survival of 20-28 weeks old fetuses is exceedingly rare. Analysis of the mechanisms of, and the conditions which permit transplacentar exchanges is fundamental for the study of the technical problems inherent in the development of an artificial placenta. Presently, extra-corporeal maintenance cannot be continued more than five days. Nevertheless, the above analyses provide directional guidelines for the pursuit of this problem.


[Non-specific fetal infections: possibilities and value of early diagnosis]. Notes of 4,000 obstetric and neo-natal cases have been analysed. The role of non-specific fetal infection (which means excluding syphilis, toxoplasmosis and German measles) has been defined with the backing of histopathological, bacteriological and clinical findings. Its role in perinatal mortality and neo-natal morbidity has been outlined. Possibilities for prevention of the condition are suggested, dealing with premature rupture of the membranes and clear-cut maternal infection. "At risk of infection" is the term that has been defined for neonates by studying carefully obstetrical clinical parameters where they are isolated or associated with other conditions but relatively innocuous. When the condition has been defined in particular cases, intensive supervision of the children at risk must be carried out so that as a result of bacteriological findings therapy can be started earlier, better adjusted to the condition and promising a better prognosis.


[The effects of cyproterone acetate on plasma testosterone in the hirsute woman]. It seems that the principal antiandrogenic action of cyproterone acetate is on the site of the androgenic receptors. Since recent work proves that there is an action on plasma testosterone, we wanted to check in women with raised testosterone levels if cyproterone lowered these levels. The answer is : Yes. Observations seem to confirm that cyproterone acetate causes a significant lowering of plasma testosterone. The basic level of testosterone and the drop in blood levels of testosterone after the antiandrogenic action has occurring do not seem to be significantly different whether the virilism is caused by an adrenal mechanism or by an ovarian mechanism.


[The descendants of 47, XYY subjects. Apropos of a family]. Having come across a family where the wife of a father who had 47 XYY chromosomes had 5 pregnancies, of which 3 were pathological (one case of anencephaly and spina bifida, one early abortion, one stillborn girl) the authors reviewed the literature concerning the lineage of parents with 47 XYY chromosomes. We have details of a total of 34 subjects who had this caryotype. 10 of them were sterile (30 per cent) and 10 (30 per cent) had a pathological lineage. Only 14 subjects (40 percent) gave rise to a line of normal descendants. The authors discuss the mechanism by which this caryotype (47 XYY) might be transmitted. They point out that so far only few cases have been published and conclude that it is important that many details about this subject should be published so that worthwhile genetic counselling will later be able to be carried out.


[Estrogen receptors in uterine fibromyomas]. The authors have sought the existence of cellular hyper-receptivity in susceptible cells in order to explain the transformation of normal uterine muscle fibres into tumours, because of the absence of true biological proof of hyperoestrogenization in fibromyomata. The total of uterine receptors for oestradiol (the total number of accessible receptors or those in which endogenous oestrogens had been taken up) were measured in normal myometrium and in fibroids removed at myomectomy or at hysterectomy. The oestradiol receptors are significantly raised in fibromyomatous tissue when the results are expressed as femtomoles per mg. of protein. All the same, there is no significant difference to be found when the results are expressed in micrograms of DNA, which leads to the supposition that there is no significant difference in the concentration of oestrogen receptors in the two types of cells. The cause of the cellular multiplication of fibromyomatous tissue should therefore be sought at another level.


[Removal of Douglas's pouch - a review of 50 cases]. The authors demonstrate the innocuousness of the method of removal of the Pouch of Douglas after a critical analysis of 50 cases followed up for between 6 months and 5 years. They believe that simple removal of the pelvic peritoneum is sufficient to overcome the condition of a pathological fold of peritoneum, but not sufficient on the other hand to correct the condition of retroverted uterus because two new peritoneal fossae form lateral to the viscerae easily because of the morphology of the pelvic cavity. They therefore stress that, at the time of the operation, it is important to suture the utero-sacral ligaments together in the sagittal plane.


[Fetal and neonatal supraventricular paroxysmal tachycardias]. The authors report 3 cases of fetal supraventricular tachycardia which carried on into neo-natal life. In one case the tachycardia was discovered in the 35th week of amenorrhoea and was able to be followed up to term. In this respect the means of making a definite diagnosis are as follows : a cardiotocogram when one happens to be able to observe the beginning or the end of an attack (in which case a definite "plateau" shape is found) and especially a fetal electrocardiogram obtained transparietally in pregnancy or by means of an internal electrode in labour. Supraventricular tachycardia is usually of marked degree and comes in isolated episodes and is not to be confused with tachycardia that is found accompanying fetal distress, occurring in pathological pregnancies. The obstetrician can therefore delay while carrying out intensive supervision of the case, which is particularly directed to searching for signs of generalised feto-placental oedema. After birth the child should be transferred to an intensive care unit where the tachycardia, if it persists, should be reduced in order to avoid serious asystolies. The outlook for the baby if there is no congenital malformation is good, since the trouble with the cardiac rhythm is usually a reflection of lack of maturity in the cardiac conduction tissues.


[Nonsurgical diagnosis of breast diseases. Hierachy of the examinations]. Systematic senological examination which includes clinical, radiological, thermographic and cytological examination of 306 breast cancers and 776 benign breast conditions has led to the ability to demonstrate the value of classifying the information obtained by these examinations. The most significant results were obtained when cytological and rediological examinations were considered together. The combination of bloodless diagnosis is improved without having to resort to negative histological biopsies. This classification should also take into account the clinical examination and thermography, which is particularly useful in screening for diseases of the breasts. Mammary echography is still another method by which the precision of the diagnosis can be improved. Thermography, using liquid crystals, is a useful complement to a first clinical examination, but is not highly reliable.


[Personal experience with salpingoplasty. Apropos of 285 cases]. 285 cases of salpingoplasty are reported and studied. Only 172 patients could be followed up recently. The percentage of pregnancies was 27-7, 20 per cent were intra-uterine pregnancies. The percentage of successful pregnancies was 45-93 if only those patients who were seen again were reviewed. The results are best when the operation is carried out in the most conservative and most anatomically restorative fashion. This fine surgery, which must be carried out so delicately and with such detail, which will appeal more and more to modern technicians using micro-surgery, should stay in the hands of specialists because there is no doubt that the level of success rises with the experience of the operator.


[Indication and technic of tubal sterilization by vaginal route. Posterior transverse colpotomy]. 40 tubal sterilisations have been carried out in 5 years. The indications most often are for grand parity. The anatomical criteria for effective sterilisation by the vaginal route should be sought by gynaecological examination under general anaesthetic. The date for the operation, the technique and its difficulties have been described. The operation lasted on an average 30 minutes. The average stay in hospital was 8.2 days. Only one local complication using this techinque was noted. On one occasion the authors had to abandon the vaginal route to reach the tubes. This simple procedure without any aesthetic inconvenience seems to them to be the ideal route for tubal sterilisation.


The hospital support game revisited.

The hospital support game revisited. This paper updates a well-known case study in which Ray Elling examined the failure of community health planning in a medium-sized American city. During the period described by Elling, the city's hospitals had managed to resist efforts to rationalize the system by marshalling the support of their respective constituents. This political gamesmanship gave way in the late sixties and early seventies to greater cooperation and a significant consolidation of the hospitals. The authors hypothesize that an important factor influencing this development was the change in medical technology. The effects on the community's health care delivery are also discussed.


Radioimmunocytochemistry-a novel immunocytochemical principle. Currently available immunocytochemical techniques rely on the detection of the antibody by means of direct labeling or indirect immunologic procedures. To increase immunocyto-chemical specificity a new radioimmunocytochemical method has been developed. In the radioimmunocytochemical method radiolabeled antigen is incubated with surplus amount of antibody. In this way only one combining site of the antibody will bind the labeled antigen, leaving the other site free to react with tissue-bound antigen. The site of reaction with tissue antigen is revealed by autoradiography. The radioimmunocytochemical method is specific and sensitive and can be combined with conventional staining methods or with immunoperoxidase techniques. It may also be useful for ultrastructural and quantitative immunocytochemistry.


The use of ruthenium and p-phenylenediamine to stain cartilage simultaneously for light and electron microscopy. p-Phenylenediamine was incorporated during the dehydration of cartilage blocks fixed with ruthenium red, glutaraldehyde and osmium. The addition of p-phenylenediamine permitted the visualization by light microscopy of cellular detail not obtained with ruthenium red alone. Ruthenium red is a widely used method for retaining and localizing proteoglycan for electron microscopy but its inability to produce sufficient density within the cells for light microscopy prevented simultaneous visualization by these two modes. The addition of p-phenylenediamine staining overcame this shorcoming without interfering with the ultrastructural detail and both stains can also be used before the making of radioautographs.


Peroxidase and gold complexes of lectins for double labeling of surface-binding sites by electron microscopy. Double labeling experiments were performed for visualization of the binding sites of Concanavalin A and anti-AHel (the lectin from Helix pomatia). The anti-AHel was labeled with the colloidal gold whereas the membrane bound Concanavalin A was demonstrated by an affinity technique using horseradish peroxidase. The two markers used could be clearly distinguished electron microscopically. The specificity of the cell surface double labeling was demonstrated in the control experiments. A topological distinct localization of the both lectin-binding sites is evident.


Cytochemical and biochemical latency of alkaline phosphatase in granules of neutrophilic leucocytes. Biochemical and cytochemical latency of alkaline phosphatase activity was demonstrated in periotoneal guinea pig leucocytes. By incorporation of a cell homogenate containing intact granules or a cell sonicate into polyacrylamide films, cytochemical latency up to 75% could be demonstrated.


Concurrent cytochemical localization of adenylate cyclase and peroxidase in the developing rat submandibular gland. An electron microscopic histochemical technique for the concurrent localization of adenylate cyclase and endogenous peroxidase is described. The procedure involves incubation of glutaraldehyde fixed tissue in adenylate cyclase medium followed by washing and incubation in 3,3'-diaminobenzidine tetrahydrochloride medium to demonstrate peroxidase activity. Adenylate cyclase was localized at the cell surface of the tissue investigated (20 day fetal rat submandibular gland) while peroxidase was localized in the rough endoplasmic reticulum and secretory granules of some cells. Biochemical and histochemical controls indicate that the procedure is valid. The potential use of this procedure and variations of the procedure are discussed.


Nucleoside diphosphatase in the onion root tip. II. The effect of inhibitors on isoenzymes. An electrophoretic study was performed to determine the number of isoenzymes of nucleoside diphosphatase in onion root extract. Five bands exhibiting nucleoside diphosphatase activity were detected when gels were incubated with inosine diphosphate, uridine diphosphate, guanosine diphosphate or cytidine diphosphate as substrates. These consisted of a single fast migrating band (band one), a group of three intermediate migrating bands (bands two, three and four) and a single slow migrating band (band five). Gels incubated with adenosine diphosphate, thiamine pyrophosphate, inosine monophosphate, guanosine monophosphate and cytidine monophosphate showed only two bands (bands one and five). Inhibitor studies showed that sodium fluoride inhibited bands one and five but not bands two, three and four. Conversely, 1% (v/v) glutaraldehyde inhibited bands two, three and four but did not inhibit bands one and five. These results suggest that two separate groups of onion nucleoside diphosphatase isoenzymes occur which have different substrate specificities and are selected against by certain inhibitors.


An ultrastructural method for the use of polyvinylpyrrolidone and dextrans as electron opaque tracers. A simple method has been developed whereby polymers of polyvinylpyrrolidone and dextran can be visualized at the ultrastructural level by postfixing tissue with osmium tetroxide partially reduced by potassium ferrocyanide. Further enhancement of polymer electron opacity is obtained by counterstaining thin sections with bismuth subnitrate chelated by alkaline tartrate. These tracers remain adequately dispersed in serum and, depending on molecular size and concentration, are easily detected at the ultrastructural level within vascular lumena, basement lamina, apical vacuoles of proximal and distal tubules and endothelial vesicles. This technique offers the potential of studying nephron and cell permeability with families of tracers, the members of which are similar in chemical affinities, charge density and molecular shape.


Sexual preference, sex role appropriateness, and restriction of social access. This study related components of gender inappropriateness that isolate target persons from sources of reward and hence function as social controls against such deviance. The study confirmed that a significant portion of negative responsiveness toward homosexuals results from the belief that their behavior is incongruent with their anatomical sex. Further, it was found that people are tolerant of out-of-sex-role characteristics when such traits are irrelevant to effective functioning in the situation, as long as the target does not display additional gender incongruence, namely, homosexuality. The results did not support the hypothesis that negative reactions to gender incongruence occur as a function of personal threat to one's own sexual identity.


Erotic imagery and self-castration in transvestism/transsexualism: a case report. After nearly 30 years of marriage, a 51-year-old man castrated himself in order to fulfill a long-standing fantasy of being a girl. There was a prior history of cross-dressing since childhood, and a reversed erotic role imagery during coitus. There was no history of schizophrenia or psychotic depression. Pair bonding with the wife was very strong. It led the patient to elect low-dose maintenance on androgen so as to permit some degree of continued marital sex. Otherwise the patient would have preferred estrogenization and, perhaps, eventual sex reassignment. The rehabilitative program as a transvestite man has continued for 3 years.


Sexual molestation and rape reported by homosexual and heterosexual women. Questionnaire returns for 225 homosexual and 233 heterosexual adult women revealed 115 cases of rape or attempted rape. Data from 78 follow-up questionnaires are summarized. About the same proportion of lesbians and heterosexuals, age 16 or over, were the object of rape. The lesbians tended to reject all men as sexual partners or companions, while the heterosexuals tended to blame themselves for not being careful. Some of the heterosexual women, age 16 to 18, interpreted the rape as a compliment to their sexual attractiveness. Girls under 16 whose assailant was a stranger are, as adults, as frequently homosexual as heterosexual. Sixteen of the 17 girls, age 4 to 16, molested or seduced (6 for a long time) by a relative or close family friend are lesbians as adults. The subjects' attitudes about the incient were highly determined by parental reactions. Inferences are drawn about the rapists' attitudes toward women.


A critique of anthropological research on homosexuality. Since the 1970 resolutions of the American Anthropological Association, encouraging more research activity among anthropologists on the topic of homosexuality, there has been less than enthusiastic response. Rather than directly attempting to provide reasons for this research failure, this paper takes a look at what actually has been done by anthropologists with an eye to assessment of their major contributions. Thus, summarized are the studies on the role of the "berdache" in primitive cultures, with a critique of the terminological problems associated; a review of some of the ethnographic accounts of homoerotic behavior among primitive folk, with comments on the weaknesses of such treatments; and, finally, a discussion of the current typological approach to the study of the homosexual community, with its multimodal rather than unimodal emphasis. In short, this is a review article that tries to assess the impact of anthropological research for the ultimate understanding of this facet of humankind.


Diabetes in pregnancy.

Diabetes in pregnancy. Diabetes, while at one time a rarity in pregnancy, is now one of the more common medical problems encountered by the obstetrician. Immaculate control is essential and can only be accomplished by close co-operation between the patient and all her medical attendants. The patient can then anticipate an almost trouble free pregnancy and should deliver a baby of normal weight with none of the usual stigmata associated with babies of diabetic mothers.


Effect of cyclophosphamide on interstitial nephritis and tubule cell proliferation in NZB/NZE mice. The effect of cyclophosphamide on renal interstitial mononuclear cell infiltration and renal tubule cell (TC) proliferation has been examined in (NZB x NZW)F1 hybrid (B/W) and control mice. TC proliferation was measured by tritiated thymidine (3/Tdr) injection, autoradiographic examination of kidney sections, and enumeration of labeled tubular cells. Cyclophosphamide administered 2 months before sacrifice virtually obliterated interstitial infiltration and significantly decreased tubule cell proliferation at 3, 5, 7, 9, and 12 months of age.


Determination of e antigen and antibody to e by means of passive hemagglutination method. A simple and sensitive method was developed for the detection of e antigen and antibody to e (anti-e) by means of the passive hemagglutination (PHA) method. Sheep erythrocytes were fixed by glutraraldehyde and coated with either e antigen or anti-e which had been specifically purified from plasma of asymptomatic carriers of hepatitis B antigen by affinity column chromatography, and then by gel filtration on Sephadex G-200. A gain in specific activity of more than 2000-fold for e antigen and 100-fold for anti-e was achieved as compared with the original plasma pool which was used as the source for purification. The PHA method was at least 300 times more sensitive than the conventional immunodiffusion method both for e antigen and anti-e assays. When the PHA method was applied for neat serum samples of 80 asymptomatic carriers, e antigen was found in 21.3% and anti-e in 62.5%, at rates much higher than those determined by immunodiffusion for the identical serum samples at 3-fold concentration (16.3 and 30.0%, respectively).


Two cytotoxic cells in peritoneal cavity of virus-infected mice: antibody-dependent macrophages and nonspecific killer cells. Two new types of cell-mediated immune cytolysis of togavirus-infected cells are compared. The peritoneal cavity of mice 2 days after infection contains a nonadherent, non-phagocytic, non-O-bearing, trypsin-resistant, EDTA-sensitive cell displaying broadly specific cytotoxicity for uninfected or virus-infected syngeneic or xenogeneic cell lines. Peritoneal macrophages from normal mice are cytotoxic to infected target cells sensitized with minute amounts of homologous antiviral IgG.


Mediation of CRP-dependent phagocytosis through mouse macrophage Fc-receptors. The purified acute phase serum constituent, C-reactive protein (CRP), was examined for its ability to interact with mouse macrophage Fc-receptors when serving as an opsonin for C-polysaccharide (CPS)-coated erythrocytes (E.CPS). The ingestion of E.CPS-CRP by macrophage monolayers was dependent on the presence of CRP on the erythrocyte surface and on the treatment of the CRP-sensitized cells with human complement (C). The ingestion of E.CPS-CRP-C was inhibited by exposing the macrophages to either 2-deoxyglucose, a selective inhibitor of Fc-receptor activity, or to aggregated gamma-globulin. Inhibition was also brought about by plating macrophages on glass-bound immune complexes of BSA/anti-BSA (IgG) but not on complexes formed with the F(ab')2 fragment of the anti-BSA. Substrate-bound complexes of CPS-CRP selectively inhibited the uptake of IgG-coated E, EA(IgG); the inhibition was proportional to the concentration of CRP used to form the complex. The opsonin-independent ingestion of latex particles was not altered by these Fc-receptor blocking procedures. These findings coupled with the recent demonstration of a sequence homology between CRP and the CH3 domain of human IgG lead us to propose shared opsonic functions for antibody and CRP.


Increase in titer of the naturally occurring human antibody to neuraminidase-treated lymphocytes after influenza. Nine of 25 (36%) humans suffering from naturally acquired influenza A infection developed significant increases in the titer of a "naturally" occurring antibody to neuraminidase-treated human lymphocytes. Only two of 43 normal and noninfluenza respiratory infection controls showed titer changes of this antibody, p less than 0.001. The antibody was not directed at influenza virus C fixation, hemagglutination inhibition, or neuraminidase antigens. Three of 10 normals given a highly immunogenic, formalin-killed influenza A vaccine developed significant titer rises. These results suggest that influenza virus, live or dead, can provoke an increase in antibody to a cross-reacting antigen present on neuraminidase-treated human lymphocytes.


Allogeneic pregnancy as immunoabsorbent. Experiments in the past have indicated that both human and rodent females undergo immune recognition of paternally derived antigens during pregnancy. Although the mechanisms that allow pregnancy to proceed in the face of this process have not yet been clarified, one simple possibility is that the paternal strain antigens are presented in a manner capable of neutralizing the maternal immune response. This predicts that allogeneically pregnant females should be capable of neutralizing passively administered cytotoxic anti-paternal strain antibody, with no consequent harm to the fetus. In this paper we demonstrate by two different methods that allogeneically pregnant mice rapidly absorb passively administered antibody directed against paternal strain major histocompatibility (H-2) antigens when compared to control mice pregnant by syngeneic or third party males. The possible sites where this absorption occurs are discussed.


Reconstitution of C1 in native proenzyme form and its use in a quantitative C1 activation test. C1 was reconstituted in macromolecular proenzyme, nonactivated form by incubation of highly purified C1q, C1r, and 125I-C1s together in the presence of calcium. C1 reformed in this manner had an equimolar ratio of C1 subcomponents, as is found in serum, and sedimented in sucrose density gradients with the 16S rate characteristic of C1 in serum. Reconstituted C1 was activatable as shown by cleavage of the 87,000 dalton polypeptide chain of C1s into disulfide linked subunits of 59,000 and 28,000 daltons, respectively, after incubation with aggregated IgG. The extent of activation may be quantitated. Reformed activatable proenzyme C1 can be used to quantitatively assess the C1-activating properties of various substances in addition to its use in the analysis of the C1 activation process.


Measurement of IgD by a double antibody radioimmunoassay: demonstration of an apparent trimodal distribution of IgD levels in normal human sera. A double antibody radioimmunoassay (RIA) was developed to measure IgD in serum and secretions. One IgD myeloma protein was used as radiolabeled antigen and standard with antiserum to a second IgD myeloma protein. The IgD standard and normal sera yielded parallel inhibition curves in the RIA and inhibition was produced by IgD and not by any of the other immunoglobulins. The assay had a lower limit of sensitivity of 0.01 International Unit (I.U.)/ml and modifications increased the sensitivity to 0.0008 I.U./ml. Measurable IgD levels were found in all 112 normal adult sera assayed (geometric mean 13.0 I.U./ml, arithmetic mean 30.1 I.U./ml, median 14.8 I.U./ml, range 0.10 to 202 I.U./ml, 95% of values between 0.19 and 156 I.U./ml. The distribution of IgD in the 112 normal sera appeared trimodal with modes at approximately 0.25 I.U./ml, 5I.U./ml, and 35 I.U./ml. IgD was measurable in nasal and bronchial washes and human milk, but could not be detected in parotid fluid.


Synthesis and release of thymidine by macrophages. An inhibitor of DNA synthesis and cell proliferation found in macrophage supernatants was subjected to further analysis. The following conclusions were drawn: a. Biochemically, the inhibitor appeared to be thymidine, judging from the migration in three different thin-layer systems and on gel filtration. b. The inhibitor was shown to be synthesized de novo by macrophage monolayers pulsed with a radiolabeled precursor of thymidine such as 14C-formate. c. The amount of inhibitor present in tissue culture fluids proved to be sufficient to block the growth of the EL-4 leukemia cell line in a manner that could be prevented with the addition of 2'-deoxycytidine. The EL-4 line was considerably more sensitive to thymidine blockade than a number of different cells tested.


Influence of inhibitors of cellular function on chemotactic factor-induced neutrophil aggregation. Chemotactic factors which induce polymorphonuclear neutrophils (PMN) to migrate directionally and release granular enzyme constituents also induce these cells to aggregate. The potency of these factors in inducing aggregation closely parallels their chemotactic and enzyme-releasing potencies. Several reagents known to influence the migratory and degranulatory response of PMN to chemotactins have been examined for their influence on chemotactic factor-induced aggregation of PMN. We have found that ambient temperatures below 37 degrees C, deoxyglucose, and iodoacetic acid inhibit PMN aggregation, whereas sodium cyanide and dinitrophenol have no effect. Inhibitors of microtubules (colchicine and vinca alkaloids) and of protein synthesis (cyclohexamide) had no effect. Cytochalasin B markedly enhanced aggregation. We conclude that chemotactin-induced aggregation is similar to the other chemotactin-induced PMN functions in the requirements for proper temperature and intact glycolytic pathways; in contrast, however, and intact cytoskeletal microtubular system appears unessential for this response. This may be explained by assuming that the chemotactic factor-induced aggregation of PMN is predominantly a surface membrane-dependent phenomenon.


Soluble factors in tolerance and contact sensitivity to DNFB in mice. II. Genetic requirements for suppression of contact sensitivity by soluble suppressor factor. Active suppression of contact sensitivity to DNFB in mice is mediated by a soluble suppressor factor (SSF) which is released in vitro by cultures of LN cells that contain antigen-specific suppressor T cells. For SSF-mediated suppression to occur, identity is required among genes in the H-2 complex between the donor of SSF and the donar of DNFB-immune LN cells. Identity in either the left half or right half of the H-2 complex was found to be both sufficient and required. Additional experiments using congeneic strains with intra-H-2 recombinants showed that the genetic homology which was required mapped to genes contained in the H-2K and/or H-2D regions of the H-2 complex.


Complementation between genetic variants affecting the response of chicken leukocytes to concanavalin A. Birds of the partially inbred G-B1 chicken line can be classified as either high or low responders to Con A, on the basis of the amount of 3H-thymidine incorporated by Con A-containing cultures of their peripheral blood leukocytes. The pattern of inheritance of the high and low responder traits suggests that the variation in response is due to genetic polymorphism at a single autosomal locus. However, the allele responsible for the low responder trait of the G-B1 line is not identical to the allele of the previously described Mr1 locus carried by the inbred low responder CC line, since (CC x G-B1 low responder)F1 birds are uniformly high responders to Con A. The responses to Con A of mixtures of CC and G-B1 low responder cells are not significantly higher than the responses of either component of the mixture alone. Thus, the gene products responsible for the complementation observed in F1 hybrids cannot complement extracellularly, and are not readily transferable from one cell to another.


Evidence for adsorption of heterospecific host immunoglobulin on the tegument of Schistosoma mansoni. Mice infected with Schistosoma mansoni were immunized against human type B Rh-positive (B+) RBC, bovine serum albumin, or horseradish peroxidase. Adult parasites, recovered by perfusion, extensively washed, and incubated in their respective antigens, selectively bound to their tegumental surfaces only those antigens to which their murine host had been immunized. All controls supported the specificity of those reactions leading to the conclusion that adult S. mansoni in mice have the ability to adsorb heterospecific antibody onto their tegumental surfaces. These surface immunoglobulins were lost within 10 min when complexed with theri antigens or within 2.5 hr when incubated at 37 degrees C. Parasites that had lost their tegumental immunoglobulins regained them when incubated in normal mouse or rat anti-human type B Rh-negative (B-) RBC serum. Those parasites that had their surface immunoglobulins reconstituted with rat anti-human B- serum specifically bound human B- RBC, suggesting the possible presence of Fc receptors on adult S. mansoni.


A new method for evaluation of intracellular inhibition of multiplication or killing by mononuclear phagocytes. A new method for evaluation of the capacity of mononuclear phagocytes to inhibit intracellular microorganisms is described. The system provides a means for assessing this effect of macrophages without concern for multiplication of extracellular organisms, effect of antibiotics, and the potential observer bias which may result from visual evaluation. It involves measurement of amount of [6-3H]UdR incorporated by Toxoplasma gondii. Differences between uptake of [6-3H]UdR by infected and uninfected macrophages can be augmented by cytosine arabinoside as this agent inhibits macrophage DNA synthesis but does not substantially alter DNA synthesis by the test organism, T. gondii.


Genetic analysis of generation of serum interferon by bacterial lipopolysaccharide. Administration of bacterial lipopolysaccharides (LPS) (LPS) to mice causes interferon (IF) to appear in their blood stream. The generation of this serum IF is "early" and its maximum concentration appears 2 hr after an inoculation of a single optimal dose of LPS (100 microgram). Two inbred strains of mice differing in their ability to generate LPS-induced serum IF were used: a high responder strain (C3H/eB), which generates high levels of IF (about 100 IF units/ml serum), and a low responder strain (C3H/HeJ), which fails to generate any detectable amounts of IF in the serum. A genetic analysis was carried out and circulating IF production was determined in the F1 hybrid, in the F2 generation, and in the progeny of backcrosses of F1 hybrids to either parent strain. The results indicate that a single dominant autosomal gene is responsible for the difference in circulating IF production between both parent strains. In addition it was shown that this type of IF is stable to treatment at pH 2 for periods of time as long as 48 hr, but is labile to heating at 56 degrees C for 30 min.


Histocompatibility-2 system in wild mice. V. Serologic analysis of sixteen B10.W congenic lines. Sixteen B10.W congenic lines carrying on C57BL/10Sn or B10 background H-2 haplotypes extracted from wild mice are described. The lines were tested serologically in two ways: first by antisera against known H-2K and H-2D private antigens of inbred strains, and second by antisera made in inbred strains against antigens carried by the B10.W lines. Both direct cytotoxicity and absorption tests were used. The analysis resulted in the serologic identification of the H-2 haplotypes carried by the 16 B10.W lines. Lines B10.STA10 and B10.STA12 are serologically indistinguishable; so are lines B10.KPA42, B10.KPA132, and B10.SNA57, whose H-2 haplotypes resemble the H-2v haplotype of B10.SM; all other B10.W lines are different from one another and different from all inbred strains known. In total, 10 new H-2 haplotypes (H-2w11 through H-2w20) and 16 new H-2 antigens (H-2.110 through H-2.125) were identified. One haplotype was discovered carrying new combination of known H-2 antigens: H-2w19, which is H-2K.19, H-2D.2. Known H-2 antigens in combination with new ones were discovered in haplotypes H-2w8 (H-2.23 and 110), H-2w12 (H-2.23 and 112), and H-2w11 (H-2.26 and H-2). The first two of these three haplotypes are probably the result of intra-H-2 crossing over that occurred during the production of B10.W lines; the last one (and the H-2w19 haplotype) is probably a natural recombinant.


The first complement component: evidence for an equilibrium between C1s free in serum and C1s bound in the C1 complex. An equilibrium between free C1s and C1s bound in macromolecular C1 exists in human serum. This equilibrium can be utilized to incorporate radioiodinated C1s into serum C1. Human sera were incubated for 40 hr at 4 degrees C with 125I-C1s to allow the exchange between free and bound C1s to reach equilibrium. The C1 complex labeled in this manner was separated from the majority of serum proteins by centrifugation in linear 10 to 30% sucrose density gradients. The resulting fractions containing 125I-C1 can be used directly and conveniently in C1 activation assays that detect the cleavage of proenzyme C1s. Electrophoretic analysis on polyacrylamide gels showed the presence of only proenzyme 125I-C1s in serum C1, whether or not the applied labeled material contained 125I-C1s or other labeled proteins. The inability of C1 to incorporate C1s was shown to be the result of decreased stability of C1 upon activation.


Immune response to phosphorylcholine. III. Requirement of the Fc portion and equal effectiveness of IgG subclasses in anti-receptor antibody-induced suppression. Anti-idiotypic antibody raised against the BALB/c myeloma proteins TEPC-15 or HOPC-8 inhibits induction of the immune response to phosphorylcholine (PC). (Fab')2 and Fab' fragments from anti-idiotype serum were prepared and anti-idiotypic antibody was purified by absorption and elution from idiotype or anti-IgG subclass immunoabsorbents. The parent material, the purified anti-idiotypic antibodies and the Fab' fragments were assayed for: i) binding to the idiotype by using a sensitive radioimmunoassay, and ii) specific suppression of the response to PC in vitro. The correlation of binding to suppression was very similar for anti-idiotype serum and both kinds of purified antibody. However, the Fab' fragments only bound to the idiotype and did not suppress. These findings indicate that the Fc portion of anti-idiotypic anti-receptor antibody (ARA) is essential for inducing suppression but that ARA-induced suppression is independent of the IgG isotype. It is postulated that idiotype suppression is mediated by the interaction of anti-idiotype with the antigen receptor and the Fc receptor thereby cross-linking both receptors.


Linkage of guinea pig Bf and C4 TO THE GPLA. Inbred and partially inbred strains of guinea pigs were typed for their polymorphic Bf and C4 patterns. In crosses and backcrosses of inbred strains of guinea pigs the inheritance of Bf, C4, and GPLA phenotypes was studied. The presented data indicate linkage of the structural Bf and C4 loci to the major histocompatibility complex (MHC) of the guinea pig, the GPLA system. Evidence for a GPLA-B-Ia crossover was obtained. Bf and C4 typing contributes to the characterization of the GPLA region in some inbred strains thus providing a simple method for determining the state of inbreeding on the one hand and providing the necessary third or fourth region within the GPLA for the detection of informative meiosis, on the other.


Partial purification of human lymphocyte-activating factor (LAF) by ultrafiltration and electrophoretic techniques. Lymphocyte-activating factor (LAF) has been produced by culturing human peripheral blood leukocytes in the presence of lipopolysaccharide and autologous human serum. The major LAF activity, identifiable by Sephadex chromatography (m.w. 13,000) was separated from most serum proteins in the culture medium by ultrafiltration with a hollow fiber device. Sucrose gradient isoelectric focusing of the concentrated ultrafiltrate yielded a single peak of LAF activity with an average isoelectric point of pH 6.8. Semi-preparative polyacrylamide gel electrophoresis of the isoelectric focusing sample resulted in separation of the LAF activity from detectable amounts of serum proteins. The recovered LAF activity was estimated to be purified more than 16,000-fold and is judged to be active in submicrogram amounts.


Potentiation of the immune response to influenza virus subunit vaccines. Influenza subunit vaccines are poorly immunogenic in unprimed lower animals and man and methods were sought to potentiate the humoral response. Influenza B intact virus vaccines potentiated the antibody response of hamsters to purified vaccines containing influenza A hemagglutinin and neuraminidase subunits. The levels of anitbody induced were at least as high as those induced by equivalent doses of whole virus. Similarly, intact heterologous influenza A virus vaccine (A/Victoria/3/75 [H3N2]) potentiated the antibody response of hamsters to A/NJ/76 [Hswl NI] subunit vaccines but large doses of intact virus were required. Considerably lower doses of homologous intact A/NJ/76 [Hswl NI] potentiated the antibody response of hamsters and sero-negative people to subunit vaccines. This suggests that future influenza subunit vaccines for use in seronegative people should contain a minimal dose of whole virus vaccine sufficient to potentiate the immune response to the subunits but insufficient to be reactogenic. A synthetic water soluble adjuvant (N-acetyl-muramyl-L-alanyl-D-isoglutamine) was also shown to potentiate the immune response of hamsters to A/NJ/76 [Hswl NI] influenza virus subunit vaccines.


Genetic control of the immune response to phage fd. IV. Complementation between H-2-linked Ir genes. The immune response to bacteriophage fd has been tested in congenic resistant strains of mice. B10 mice are high responders; B10.BR and B10.D2 mice are low responders. B10.A mice with the recombinant H-2a haplotype as well as heterozygous H-2k/d (B10.BR X B10.D2) F1 mice respond with high antibody titers, indicating the existence of two complementing Ir genes. From the results obtained with recombinant and F1 hybrid mice, Irfdalpha can be localized to the right of the crossover between the IE and IC subregions, Irfdbeta to the left of IB, presumably at IA. Complementation is observed with selected pairs of alleles only, suggesting alpha-beta coupled complementation. In addition to genes in the H-2 complex, there are non-H-2 background genes modulating H-2 regulated anti-fd response. B10.BR mice lack primary IgM and IgG antibodies, yet are able to mount a secondary IgG anti-fd response, giving evidence for intact memory formation in this low responder strain. The combined data are most easily interpreted by assuming that both H- and non-H- genes modulate the effective immunogenic dose of antigen.


Basophil-derived platelet-activating factor (PAF) as an in vivo mediator of acute allergic reactions: demonstration of specific desensitization of platelets to PAF during IgE-induced anaphylaxis in the rabbit. The anaphylactic response to an i.v. injection of antigen into rabbits making only IgE antibody against the antigen was shown to be preceded by a disappearance of the metachromatic staining properties of the circulating basophils and was accompanied by marked but transient thrombocytopenia. The platelets which returned to the circulation 1 hr after the anaphylaxis were shown to be unresponsive to the secretion-inducing activity of basophil-derived platelet-activating factor (PAF) when compared with platelets examined before antigen challenge. By contrast, platelet responsiveness to other stimuli such as collagen, thrombin, and C3b was unchanged. The specific desensitization to PAF provides strong evidence for the action of this mediator on platelets in vivo during IgE-mediated anaphylaxis and provides a useful tool for detecting the effects of particular cell activators in inflammatory reactions.


Intravascular aggregation and pulmonary sequestration of platelets during IgE-induced systemic anaphylaxis in the rabbit: abrogation of lethal anaphylactic shock by platelet depletion. Significant alterations in the circulatory properties of platelets have been documented during IgE-induced systemic anaphylactic shock in the rabbit. Within 30 to 60 sec after i.v. antigen challenge, platelet aggregation occurs in both the venous and arterial circulations. The platelet aggregates then sequester in small blood vessels of various organs, particularly in the lung. The organ sequestration results in the development of a profound thrombocytopenia within 3 to 5 min after antigen challenge. Fifteen min later deaggregation of platelets occurs and the platelets return to the peripheral circulation within normal, prechallenge levels by 60 min. Additional experiments demonstrated that platelet depletion before antigen challenge abrogates the lethal effects and significantly reduces the pathophysiologic manifestations of IgE-induced systemic anaphylaxis. We conclude that the IgE-induced platelet alterations, probably induced by the intravascular release of basophil and perhaps mast cell-derived platelet-activating factor (PAF), play a major role in the pathogenesis of systemic anaphylaxis in the rabbit.


Apparent improvement of antisera for radioimmunoassay by treatment with sodium iodide. The concept that antisera for radioimmunoassay can be improved by treatment with NaI and dialysis is challenged. Some antisera are little affected by NaI. Even when NaI does dissociate endogenous antigenic material from crude antiserum, in the subsequent dialysis NaI is dialysed at a much faster rate than antigen, allowing reassociation to occur. The reported improvement of antisera by the NaI method cannot be attributed to the effect of NaI but can be explained by the dilution which occurs during dialysis.


Mechanization of the enzyme-linked immunosorbent assay (ELISA) for large scale screening of sera. An on-line routing system (including dispensers, washing device, spectrophotometer, carts, elevators and identification device) for macro-ELISA is described. The system enables processing of 4,000 sera daily by 2 persons. Test results are presented on a data sheet as a combination of identification number and extinction value.


A simple technique for studying immunoglobulin synthesis by normal and malignant plasma cells in vitro. Plasma cells from human marrows are saturated with C-14 labelled amino acids, harvested and recultured in unlabelled growth medium. The appearance of radioactivity in the growth medium then provides a simple and rapid measure of protein synthesis. The secreted radio-labelled material is characterized by isoelectric focusing and autoradiography in acrylamide gels, a technique which has advantages over established serological methods.


Comparative study of macrophage migration in different strains of mice: absence of migration in C3H mice. In the course of studying the susceptibility of mice to Salmonella infection, we discovered that C3H/HeJ mice differ from other strains in both their susceptibility to infection and their response to prior vaccination. Others have noted the absence of the usual B cell stimulation by LPS in these mice. To investigate the status of delayed hypersensitivity in C3H/HeJ mice, we tried to use the technique of macrophage migration. Under conditions of normal migration for other strains, C3H/HeJ peritoneal and splenic macrophages failed to migrate, thus establishing another differentiating immunological trait for this strain.


Characteristics of soluble immune complexes prepared from oligovalent DNP conjugates and anti-DNP antibodies. The stability of soluble immune complexes was investigated after isolation by gel filtration and sucrose gradient ultracentrifugation. Soluble immune complexes were formed between specific goat anti-dinitrophenol (DNP) antibodies and DNP conjugated to a large (19 S) carrier, namely bovine thyroglobulin. The composition and molecular weight of these complexes were determined by ultracentrifugation on calibrated sucrose density gradients and the use of different isotopic markers for antigen and antibody. A good separation of immune complexes containing one, two, or three antigen molecules per complex was obtained by ultracentrifugation while gel filtration was less effective. Ultracentrifugational analysis of fractions isolated by these two procedures showed that large immune complexes containing more than one antigen were relatively labile, whereas small immune complexes containing one antigen were stable. The stability of large immune complexes was dependent on dilution. Because dilution affects the size and composition of soluble immune complexes, it is important to emphasize that for the investigation of a causal relationship between the biological properties and the size and composition of immune complexes, analysis of the immune complexes should be performed in the same dilutions in which they will be used experimentally.


A microculture system for the measurement of antigen-induced murine lymphocyte proliferation: advantages of 5% horse serum and 5 X 10(-5) M mercaptoethanol. Short term microculture systems which measure murine lymphocyte proliferative responses to mitogens are well established. We demonstrate here that these microculture methods are not suitable for antigen-induced responses because of the high levels of murine lymphocyte proliferation in control cultures associated with the use of fetal calf serum or human serum. We also show that this problem can be eliminated with the use of a combination of 5% horse serum and 5 X 10(-5) M mercaptoethanol. We describe an antigen-induced murine lymphocyte proliferation microculture system in which good stimulation indices are achieved and the lymphocyte proliferation in control cultures remain at a low level throughout the 7 day culture period.


Quantitative analysis of macrophage phagocytosis by uptake of particulate 192iridium. This study describes a method for the quantitation of macrophage phagocytosis by measuring the uptake of small particles of the radioactive inert metal, iridium (192Ir). It is shown that macrophage labeling with 192Ir was uniform, and the label was retained by the macrophages up to 21 days post-plating (when all cultures were terminated). 192Ir labeled macrophages were able to phagocytose other particles such as sheep red blood cells and were able to be activated to become tumorcidal in vitro 21 days following initial 192Ir uptake. Therefore, 192Ir could be a useful tool for the quantitative analysis and correlation of phagocytosis, tumor cytotoxicity and other macrophage functions.


An indirect solid-phase microradioimmunoassay for human IgM-anti-IgG (rheumatoid factor). In order to satisfy the general need for a more precise measurement of the serum concentration of rheumatoid factor (RF) than is presently obtainable with the latex test, we developed a rapid indirect solid phase micro radioimmunoassay for RF determination. The assay involves the binding of IgM-anti IgG (RF), to the polyclonal, native or denatured human IgG dried on the bottom surface of microtiter plates; the amount of antiglobulin bound is then determined by adding 125I-labelled goat anti-human IgM (125I-AHIgM). Variations of reagents and their concentrations, temperatures and incubation times were studied to find the optimal conditions for test sensitivity, specificity and reproducibility. The test has a within day and between day average coefficient of variation of 9 and 15% respectively, showing its advantages over the latex test. The results obtained by studying 100 human sera from patients with positive or negative latex test show that the test should prove valuable in diagnosis and research.


A simplified isotope release assay for cell-mediated cytotoxicity against anchorage dependent target cells. An assay for cell-mediated cytotoxicity has been developed in which anchorage-dependent target cells are cultured on small plastic beads in suspension. Confluent target cells on the beads are handled by methods appropriate to suspension-grown cells and labelled with chromium-51, iodine-125 and [3H]proline. Fetal human lung fibroblasts and HeLa cells were used as targets in model experiments measuring human natural killer cell activity. In 20 h experiments, chromium-51 was the most suitable isotope. In 40 h experiments, [3H]proline release assay was superior to chromium-51 and iodine-125 assays. The bead cytotoxicity assay offers a rapid and simple isotope release technique for anchorage dependent cells because no trypsinization and re-seeding of target cells is needed.


The measurement of total serum IgE levels in rats. A direct solid phase radioimmunoassay for the measurement of total serum IgE in the rat has been developed. The assay is very sensitive with a lower limit of detection of 5 ng/ml. The intra and interassay reproducibility proves to be very acceptable and the specificity is demonstrated by looking at the interferen-e by other immunoglobulins. Preliminary results on total serum IgE levels are given.


Semi-automatic solid-phase radioimmunoassay for carcinoembryonic antigen. A solid phase, double antibody radioimmunoassay has been semi-automated for the quantitation of carcinoembryonic antigens. Sepharose-bound rabbit anti-goat IgG immunoglobulin was used as the second antibody, reactions were carried out in 96-well microtiter plates, and samples processing (filtration and washing) was accomplished with the aid of a 24-sample harvester.


Isolation of soluble immune complexes by affinity chromatography using staphylococcal protein A--Sepharose as substrate. This report describes a technique for the general isolation of immune complexes, based on a combination of gel filtration and affinity chromatography. The first step is the preparation of a globulin-enriched fraction by precipitation with ammonium sulfate at 50% saturation, or of an immune-complex-enriched fraction by precipitation with 5% polyethylene glycol 6000. The enriched fraction is then subfractionated by gel filtration in Ultrogel AcA 34. The immune complexes elute close to the void volume in the macroglobulin peak, separated from monomeric IgG molecules. This peak (sometimes subdivided into two fractions) is then submitted to affinity chromatography on a protein A--Sepharose cooumn. Most immune complexes contain IgG molecules and therefore bind to the column. Almost no protein is bound when normal serum is fractionated according to this method, and no immunoglobulins are detectable in the acid-eluted fraction from the protein A--Sepharose column. In two patients with soluble immune complexes in their sera we eluted immunoglobulin-containing fractions from the column; in one, these fractions had high rheumatoid factor titers; and in the second, with a clinical diagnosis of systemic lupus erythematosus, a similar fraction contained RNA.


Particle counting immunoassay (PACIA). I. A general method for the determination of antibodies, antigens, and haptens. By using a device designed for counting blood cells, it is possible to measure the agglutination of polystyrene beads (0.8 mu) with accuracy and great sensitivity, the agglutination resulting in a reduction in the number of particles. The latter coated with antigen can be used for determining IgM or IgG antibodies e.g. human rheumatoid factor or rabbit anti-bovine serum albumin antibodies. Macromolecules with multiple antigenic determinants agglutinate particles carrying specific antibodies. This system has been applied for determining HPL and alpha1-fetoprotein with a threshold of sensitivity of about 10 microgram/1. However the agglutination was decreased by serum factors which led to a 10-fold loss of sensitivity. The interference of rheumatoid factor which agglutinated the particles coated with rabbit or goat immunoglobulins could be avoided by reduction of the serum to be analyzed with 5mM dithiothreitol for 5 min. Haptens, i.e. DNP-lysine and T4, were determined by their inhibitory activities toward their specific antibodies, the agglutinator being a hapten-macromolecule conjugate or the antibodies themselves.


An indirect anti-immunoglobulin rosetting reaction to detect alloantibodies to human lymphocytes. A test is reported which detects alloantibody, absorbed onto the surface of human lymphocytes from multiparous antisera, by means of a red cell rosette assay. The red cells, trypsinised ox, are coupled with anti-immunoglobulin using chromic chloride. The antiglobulin used is rabbit anti-human IgG (Fc), chosen to avoid reaction with the surface immunoglobulin naturally present on human B lymphocytes. The reaction is termed the Indirect Anti-immunoglobulin Rosetting Reaction (IARR). The IARR is shown to be specific in the following ways: anti-immunoglobulin coupled ox cells do not react with normal human lymphocytes nor with lymphocytes treated with non-reactive serum. Red cells coupled with normal rabbit IgG do not react with normal or alloantibody coated lymphocytes. Multiparous sera (reactive with other individuals) do not react with cells of the serum donor in the IARR. Finally, the coupled red cells do not usually react with lymphocytes which have absorbed immune complexes onto their Fc receptors. The IARR is shown to be more sensitive than a standard cytotoxic test for detection of alloantibody. Several possible applications of the IARR are discussed.


The effect of pregnancy on lymph node weight in the mouse. Lumbar and renal lymph nodes, inguinal lymph nodes and spleen weights were recorded during pregnancy in mice after mating with males of the same (syngeneic) or a different (allogeneic) strain. The weights of these nodes increased during pregnancy. Spleen weights increased up to the fifteenth day of pregnancy and then decreased in weight. There was no consistent correlation of any of these measurements with the extent of the antigenic dissimilarity between mother and conceptus.


e antigen and antibody, DNA polymerase, and inhibitors of DNA polymerase in acute and chronic hepatitis. A high positive correlation was found between e antigen (HBe Ag) and DNA polymerase in hemodialyzed patients with acute hepatitis B, chronic carriers of hepatitis B surface antigen undergoing hemodialysis, and patients with chronic hepatitis. In contrast, the correlation was poor in nonhemodialyzed patients with acute hepatitis. Among the patients with chronic hepatitis, HBe Ag and DNA polymerase were were found mostly in those with aggressive hepatitis and rarely in those with persistent hepatitis. This difference was significant (P less than 0.01) and suggests that the persistence of these antigens may be a factor in the progression of the disease. Our data also indicate that the development of antibodies to HBe Ag (anti-HBe) might be a sign of a favorable prognosis, since 50% of the patients with persistent hepatitis vs. 6% of the patients with aggressive hepatitis were anti-HBe-positive. Inhibitors of DNA polymerase, which are possibly antibodies, appeared regularly after acute hepatitis and were transient. Their presence may be associated with viral replication.


In vitro activity of five oral cephalosporins against anaerobic pathogenic bacteria. The in vitro inhibitory activities of cefaclor and and cefatrizine, two new orally absorbed cephalosporin antibiotics, against 44 isolates of anaerobic pathogenic bacteria were measured using the agar dilution procedure of the World Health Organization-International Collaborative Study. Tests also were performed with cephalexin, cephaloglycin, and cephadrine, as well as with the parenteral cephamycin antibiotic cefoxitin. Cefoxitin was the most active antibiotic and inhibited the majority of isolates at a concentration of less than or equal to 4 microgram/ml. None of the oral cephalosporins was clearly superior against all of the anaerobic isolates; only cephadrine and cefatrizine appeared to have any potential clinical value.


Clindamycin-associated colitis due to a toxin-producing species of Clostridium in hamsters. Clindamycin-associated enterocolitis in hamsters was studied to detect and characterize a transmissible agent. It was found that the disease could be transferred by cecal contents and filtrates of cecal contents (pore size of filter, 0.02 micron) obtained from animals after administration of clindamycin. Subsequent work showed that enterocolitis could be produced with broth cultures of a species of Clostridium recovered from cecal contents of animals with clindamycin-induced disease. The cell-free supernatant of this strain also caused enterocolitis. Cecal contents from animals with clindamycin-induced disease incubated with gas gangrene antitoxin failed to cause intestinal lesions. These experiments indicate that clindamycin-associated colitis in hamsters is due to a clindamycin-resistant, toxin-producing strain of Clostridium.


Mesenchymal cell atypicality in inflammatory polyps. Excised nasal polyps should be examined histologically for classification as to aetiological process. Recently a 'pseudosarcomatous' change was reported in nasal polyps. We reviewed one hundred excised polyps for mesenchymal atypicality and found a wide degree of mesenchymal atypicality in the majority of nasal polyps. Nasal and choanal inflammatory polyps are the most frequent nasal masses which the otolaryngologist has to treat. Surgical excision is frequently required to alleviate the nasal symptoms of polyps unresponsive to medical therapy. Excised nasal polyps should be examined histologically for classification as to possible aetiological mechanisms and to rule out neoplastic lesions (Shea, 1947). Smith, Echevarria, and McLelland (Smith, 1974) recently described 'pseudosarcomatous changes' in nasal polyps. This stimulated us to investigate our inflammatory nasal polyps for these histological changes. Causal observations demonstrated an impressive number of inflammatory polyps with varying amounts of atypical mesenchymal stroma cells. To quantitate these alterations, we reviewed the histological slides from one hundred patients with excised inflammatory polyps and the results are the subject of this report.


Anaesthesia of the tympanic membrane using iontophoresis. Experience is described of the use of iontophoresis of 2% lignocaine to achieve effective tympanic membrane anaesthesia. The main uses are for outpatient myringotomy and transtympanic electrocochleography. Complete anaesthesia of the drum has been obtained in all cases, and those patients who had previously had these procedures carried out without anaesthetic, or with surface 'anaesthesia' only, all stated a marked preference for iontophoresis. No complications have occurred.


Sudden deafness and facial palsy from metastatic bronchogenic carcinoma. A 52-year-old man developed sudden total bilateral deafness, and unilateral facial palsy, without other symptoms and findings. He died two months later of of bronchogenic carcinoma metastatic to dura, brainstem, pons, carebellopontine angle, cerebellum and cranial nerves III, VI, VII and VIII. There was bilateral internal auditory canal erosion. Tumour replaced right facial, acoustic and vestibular nerves. Tumour infiltrated spiral ganglion, cochlear nerve, cochlear aqueduct, and destroyed nearly all facial nerve fibres to the level of the stapedius muscle. No tumour cells were found on the left, but few fibres of facial, acoustic and vestibular nerves survived. Both ears showed some cochlear outer hair cell destruction. Metastatic tumour to temporal bone or dura should be considered when loss of peripheral VIIth or VIIIth nerve function occurs.


Malignant oncocytoma.

Malignant oncocytoma. A case of malignant oncocytoma of the parotid gland in a 32-year-old male is presented. Ten months after parotidectomy an undifferentiated carcinoma, in which oncocytes still could be recognized, developed in the operated area. According to the literature available to us, this is the second reported case in which malignant transformation in a benign oncocytoma of the salivary gland has been observed.


Nasal encephalocoeles associated with unilateral absence of the cochlea. A case of nasal encephalocoele associated with unilateral absence of the cochlea is described. The association of congenital abnormalities of the skull and the spine with deformities of the inner ear is discussed together with the value of tomography in the assessment of these children.


Anterior lenticonus--an unusual feature of Alport's syndrome. A case of anterior lenticonus with haematuria and sensorineural deafness is presented. It is suggested that the anterior lenticonus forms an important clue to the diagnosis of Alport's syndrome which can be confirmed by detailed audiometric examination and urine analysis of other family members.


Prognostic features of ventricular tachycardia complicating acute myocardial infarction. Prognostic features of 115 patients with ventricular tachycardia complicating acute myocardial infarction were analyzed. Age, sex, infarct location and peak CPK levels were not significantly different when comparing survivors (S) and non-survivors (NS). Highly significant clinical characteristics of NS compared to S were: heart rate, presence of cardiogenic shock and a poor response to lidocaine therapy (P less than 0.0001, 0.0003 and 0.001 respectively). Electrocardiographic features distinguishing S and NS were: coupling intervals (S = 522.9, NS = 389.9, P less than 0.004), prematurity index (S = 1.36, NS = 1.04, P less than 0.001), ventricular tachycardia rate (S = 132, NS = 174, P less than 0.0013) and number of episodes of ventricular tachycardia (S = 4.04, NS = 6.75, P less than 0.0058). These findings have importance for the evaluation of newer active and prophylactic therapies.


Restorative effect of epinephrine on the electrophysiologic properties of depressed human atrial tissue. The effects of epinephrine on the electrophysiologic properties of human right atrail tissue, obtained at cardiac surgery, were evaluated utilizing standard microelectrode techniques. In studies of electrophysiology, epinephrine had little effect on the resting membrane potential and transmembrane action potentials of normal atrial fibers. Epinephrine enhanced phase-4 depolarization and increased automaticity in normal fibers but hyperpolarized partially depolarized atrial fibers and decreased automaticity. The hyperpolarizing action of epinephrine resulted in an increase in action potential amplitude and dV/dt and enhanced conduction. Active force increased 40-230% in depressed tissues when exposed to epinephrine. Epinephrine-induced hyperpolarization of depressed atrail fibers may have a beneficial effect on atrial arrhythmias and depressed contractility encountered clinically.


P-R interval in relation to heart rate during exercise and the influence of posture and autonomic tone. P-R interval (PR) in relation to heart rate (HR) during exercise was studied in healthy men. When subjects were in a recumbent position, mean PR between HR 90-140 beats/min (bpm) decreased linearly from 167 +/- 8 ms to 136 +/- 5 ms. (Regression line: PR = 0.287 HR + 182.9, r = 0.40). PR did not decrease further at HR up to 180 bpm. When subjects were in a sitting position, a further decrease occurred after HR 150-160 bpm. The shortest PR observed during exercise was 100 ms. The decrease of PR between HR 90-140 bpm was affected by atropine but not by propranolol. Higher HR was not achieved after propranolol, and after atropine there was no difference in PR in either exercise position compared to the two exercises without any drug. Thus, exercise induces a decrease in PR which is for the most part completed at HR 140-150 bpm and is mainly achieved by a withdrawal of the parasympathetic tone. PR at HR 90 bpm was correlated to body surface area, indicating that the PR duration is related to the body and heart dimensions.


The vectorcardiogram as an aid to diagnosis in left ventricular-right atrial communication. Frank vectorcardiograms (VCGs) in four patients with left ventricular-right atrial (LV-RA) communication were analyzed and compared with published values of VCGs of normal individuals and those from a group of patients with membranous ventricular septal defect. It was observed that the QRS-loops in the frontal and sagittal planes of the patients with LV-RA communication are shifted more superiorly than usual. Thus, almost 50% of the QRS-loop area was superior to the X- and Z-coordinates in the frontal and sagittal planes, whereas in the controls less than 10% of the frontal and sagittal plane QRS-loops were superior to the horizontal axes. The QRS-loops of patients with LV-RA communication thus seem to be halfway between normal and endocardial cushion defect loops. An unusual degree of superior orientation of the QRS-loop in a patient with clinical findings of a ventricular septal defect should arouse suspicion of a LV-RA communication. Also in patients with an isolated ventricular septal defect but with an exaggerated superior orientation of the QRS-loop in the frontal and sagittal planes, the interatrial septum should be examined at the time of operation to exclude the possibility of an associated LV-RA communication.


Endocardial electrogram amplitudes and stimulation thresholds in monopolar electrodes: possible correlations. The possible correlations between unipolar endocardial electrogram amplitudes and voltage threshold stimulation were tested with electrodes having metallic tips of two different areas. The threshold measurements were always studied with the same pulse characteristics. The data were analyzed through five mathematical functions (linear, exponential, potential, hyperbola and translate hyperbola). The results show a significant correlation, mainly by hyperbolic and exponential functions. Consequently, we could deduce a parameter value since we already knew the other one. This was possible not only during the first implantation, but also later on.


In vivo atrial excitability and anti-arrhythmic drugs. The threshold of excitability of the atrial muscle was studied in the in vivo beating canine heart. Unipolar cathodal and anodal strength-interval curves were constructed and found to be dissimilar in shape. It was found that at any interval within the relative refractory period of the atrium, as in the ventricle, there is a wide range of current levels delineated by an upper (TU) and lower (TL) limit of threshold which can stimulate the atrial myocardium. Within these limits the threshold varies spontaneously and can be reduced to TL level by a run of extrasystoles. Such TU and TL curves were repeatedly determined following administration of therapeutic doses of quinidine, procaine amide or lidocaine. It was observed that all three drugs prolonged the refractory period. The TU values increased following each of the drugs, and mostly after quinidine, while the TL curve was less affected by quinidine. It is suggested that the exit block thus produced is the principal mechanism whereby quinidine depresses atrial disrhythmias.


Direct-writing color vectorcardiograph.

Direct-writing color vectorcardiograph. By employing a specially designed instrument, a "color vectorcardiogram," a single, colored horizontal plane vectorcardiogram (VCG) was written directly on the recording paper. At first, the voltage in the X, Y and Z axes was memorized into the digital memory cells. After that, the voltage in the X and Z axes was slowly read out so that a horizontal plane vectorcardiogram was written directly on the paper of the X-Y recorder. The color of the recording pen varied according to the voltage in the Y axis, which was read out simultaneously with the voltage in the X and Z axes.


Complex atrial dysrhythmias in a patient with congestive cardiomyopathy. A patient with primary cardiomyopathy and long standing atrial fibrillation presented with atrial tachycardia and AV block. Atrial pacing sequentially converted the atrial tachycardia to atrial flutter (biatrial) and to the dissimilar atrial rhythms of right atrial fibrillation with left atrial flutter, right atrial flutter with left atrial electrical standstill and right atrial fibrillation with left atrial standstill. The clincial milieu and intracardiac recordings are presented.


Reduction of the rate of outgrowth, cell density, and cell division following removal of the apical ectodermal ridge of the chick limb-bud. Removal of the apical ectodermal ridge causes a reduction in the rate of outgrowth of the wing-bud and the loss of distal parts. More specifically it causes a short-term increase in cell density and cell death and a decrease in the rate of cell proliferation. The evidence supports the hypothesis of density-dependent control of cell division and suggests that there may also be a mechanism regulating skeletal length at the time of differentiation. An informal model is presented to explain the observations.


Hormone control in regeneration: effects of somatostatin on appendage regeneration, blood glucose and liver glycogen in Diemictylus viridescens. In the present communication, synthetic somatostatin, a hypothalamic factor which has a known inhibitory effect on the release of growth hormone, thyroid-stimulating hormone, prolactin, insulin and glucagon in man and other mammals, was found to have an inhibitory effect on limb and tail regeneration in adult Diemictylus viridescens, when the newts were treated with a daily dose of 3-5 or 15 microgram/animal for a period of 34 days post-amputation. At the higher dose, the animals exhibited total inhibition of appendage regeneration in a few cases and the remainder showed a considerable delay compared to the controls; none of the experimental animals reached the advanced four-digit stage achieved by the controls. Furthermore, the blood glucose and liver glycogen values in the somatostatin-treated animals were significantly lower than the control values. Mechanisms in the storage, mobilization and utilization of glucose (involving hormones) are discussed in relation to appendage regeneration in the newt and possible controls of regeneration at the level of the hypothalamus are suggested.


Cell growth and differentiation of murine extra-embryonic fetal and adult hematopoietic tissues in diffusion chamber cultures. Studies were conducted with diffusion chambers (DC) filled with cell suspensions from different CF1 murine hematopoietic tissues: adult peripheral blood; adult tibial marrow; day 17-5 of gestation fetal liver, spleen and thymus; day 14-5 gestation fetal liver; day 10-5  of gestation yolk sac. After an initial decrease in DC cell numbers on day 2 of culture, growth of each cell group continued, but, at different rates. ATM had the highest growth ratio and FT-D17-5(2) had the lowest. The growth rates for APB and FL-D17-5 were similar. FS-D17-5 and FL-D14-5 cultures did not recover from the day 2 values (i.e. FL-D14-5 DC values on day 13-14 of culture were half that recorded on day 2). The YS-D10-5 DC cell numbers continued to increase throughout the 14 days of study. The profile of cellular elements from the DCs did not reflect the original cell suspensions. The predominant cell type recovered from peripheral blood cultured for 14 days was the macrophage. By day 10-14 of culture, the populations of cells harvested from the fetal tissue DC groups were similar to that of tibial marrow. Both proliferative and mature granulocytes, and macrophages were the predominant cell types. The yolk-sac pattern of cytodifferentiation recorded on day 7-14 was unlike that of the other groups. These DC cultures were comprised of mainly macrophages and plasma cells.


Nuclear transplantation from intestinal epithelial cells of early and late Xenopus laevis tadpoles. The aim of these experiments was to test whether the developmental potential of nuclei from intestinal epithelial cells of Xenopus laevis tadpoles declined during the life of the functional larval gut. The results of transplantation of nuclei from three different stages of development were compared: stages 46-48, when feeding begins and while yolk is still present but before the formation of the typhlosole; stage 57, just prior to the onset of metamorphic reorganization; and stage 54, an intermediate stage. The results showed that there was no change in developmental potential of these nuclei during the life of the larval gut, thereby disproving the hypothesis that nuclear transplants from intestinal epithelial cells of early tadpoles of X. laevis will support extensive development because the cells are not fully functional. However, nuclei from the intestinal epithelial cells were less able to support development than blastula nuclei. It was concluded therefore, that the developmental potential of the gut nuclei is restricted relative to that of the blastula nuclei, but that these restrictions are reversed in a small proportion of cases.


Retinal growth in double dorsal and double ventral eyes in Xenopus. The growth of normal and surgically produced compound dorsal and ventral retinae in Xenopus laevis has been studied autoradiographically following injections of [3H] thymidine at stages 50 and 58. The animals were sacrificed 3 weeks after metamorphosis. The histogenetic pattern of the dorsal and ventral retinal halves was different at the three time points investigated, i.e. up to stage 50, between stages 50 and 58 and between stage 58 and 3 weeks after metamorphosis. Asymmetrical dorsal retinal growth occurred up to stage 50. From stage 50 onwards the retinal growth tendency reversed so that more ganglion cells were produced along the ventral than the dorsal ciliary margins. The overall preponderance of ventral retinal growth was 32.4% in cell number and 12.4% in retinal length from early embryogenesis to 3 weeks after metamorphosis. The characteristic histogenetic pattern of the dorsal and ventral retinal halves was maintained in an ectopic position in the compound eye, indicating that this particular property of the retinal halves is intrinsically determined.


The blastomere pattern in echinoderms: cleavages one to four. The results of a longitudinal study of the blastomere pattern in six embryos during the first four cleavages are reported. At each cleavage stage optical sections through an embryo, taken at vertical intervals of 5 or 10 micron, were recorded on 35 mm film: digitization of the blastomere contours and computer analysis allow calculation of the center, radius, surface area and volume of each blastomere. The subjective impression of exquisite regularity seen in normal echinoderm blastulae acquires a quantitative dimension from the present study. For example, the individual angles formed by the various quartets of blastomeres depart from right angles by at most a few degrees. The egg volume was found to be conserved up to the fourth cleavage. At the 16-cell stage, unlike the earlier stages, the blastomere positions cannot be ascribed solely to the position and orientation of the respective cleavage planes. Finally, a few features of a formal model of these early cleavages are sketched.


Variation in Habrobracon juglandis ovariole number. I. Ovariole number increase induced by extended cold shock of fourth-instar larvae. Variation from zero to five ovarioles in the braconid wasp, Habrobracon juglandis, is reported for dissections of females from laboratory stocks. Four ovarioles (two per ovary) is the standard phenotype for the species. Storage of fourth-instar larvar in a refrigerator resulted in a large proportion of females with an increased number of ovarioles (5-11). A 1- to 2-week cold exposure during the period of ovarian differentiation is required for the maximum effect.


Cell disposition and adhesiveness in the developing chick neural retina. The adhesive properties of neural retinal cells located in the ventral and dorsal hemispheres of chick embryonic eyes were investigated. Cell adhesion was monitored using the collision efficiency method; this technique provides a system in which it is possible to identify any preferential adhesions that may occur between cells. In this study no adhesive specificity was detected between cells of the dorsal and ventral retina. This evidence would not support theories which invoke preferential cell adhesion as an explanation of the ordered projection of the neural retina onto the optic tectum during development.