Evaluation of fluorochemicals for liquid membrane oxygenation. An attempt is being made to develop an oxygenator utilizing gaseous oxygen bubbles completely encapsulated with fluorochemical, thereby avoiding the detrimental changes induced by a blood-gas interface. After the feasibility of this method of oxygenation had been proved, the biocompatibility of the fluorochemicals was investigated. No significant changes in human red blood cells, fibrinogen, or platelets were induced by chronic in vitro contact with fluorochemical over a 24-hour peroid. There is no evidence that the fluorochemicals tested extract lipids from plasma. A device which allowed continuous formation of a blood-fluorochemical interface was utilized in vitro with human blood and in vivo with dogs. No significant alterations were induced by fluorochemicals in the human or animal blood or in the canine organs at autopsy. A prototype oxygenator is now undergoing evaluation. A method of analyzing for fluorochemical in blood and other protein solutions is presented.


Effect of fluorocarbon emulsions on the mechanical fragility of normal and sickle cells: in vitro studies. Mechanical fragility measurements have been made in vitro on fluorocarbon emulsions mixed with normal and sickle cells in plasma to determine the effect of fluorocarbon. Emulsions of FC-80 with Pluronic F-68 were added to give final solutions of 0, 1, 5, 10, and 20% fluorocarbon emulsion. The effect of the fluorocarbon emulsion was observed in the presence and absence of oxygen. In the presence of oxygen, there was no effect of the fluorocarbon emulsion on the mechanical fragility of normal or sickle cells. In deoxygenated systems, however, there was significantly less hemoglobin in the plasma during the mechanical fragility test with fluorocarbon emulsion added to sickle cell blood. The normal blood was not affected by the fluorocarbon emulsion in the deoxygenated system. Five percent of fluorocarbon emulsion was required for a significant effect on the deoxygenated sickle cells. Since the effect of the fluorocarbon emulsion was in a deoxygenated condition, the effect is due to the presence of the fluorocarbon emulsion and not its oxygen carrying capability.


Artificial blood and the National Blood Policy. Artificial blood will become important to the extent that it is safer, more effective, more economical, and more readily available than natural substances. If physicians would accept blood substitutes, their use might help to eliminate prevailing shortages of blood and increasing needs for plasma. The high demand for albumin, one of the safest biologics known, is pushing the requirement for plasma to higher levels every year. The albumin situation illustrates both the value placed on hemotherapeutic agents of proven safety and the problem of mobilizing sufficient blood resources for their production. The solutions to this problem and others addressed by the National Blood Policy will involve both scientific efforts and improvements in the operation and organization of blood service systems. Thus, the search for blood and plasma substitutes must be viewed as just one of a set of management and scientific approaches to meeting the need for safe, effective hemotherapuetic agents.


Food and Drug Administration regulations and licensure. The regulatory requirements of the Food and Drug Administration for products that are defined as drugs and/or biologics are extensively documented in the applicable sections of the Code of Federal Regulations. They include broad requirements for good manufacturing practices and a number of general requirements for matters such as sterility, stability, pyrogens, and labeling. In addition, for biological products, specific standards are published for comment in the Federal Register and then as final rules prior to the acceptance of license applications for new products. The major criteria for such products are safety and effectiveness when the products are used prudently as directed in accompanying circulars. It is generally sound practice to obtain extensive toxicologic and pharmacologic knowledge of potential new products prior to embarking on clinical trials. These trials must be described in investigational new drug applications (INDs), if the products will be entered into interstate commerce. Clinical trials should be planned and conducted using sound scientific and ethical principles in order to obtain objective evidence of clinical effectiveness without unduly jeopardizing the participants in such studies.


Influence of estrogen and progesterone treatment on ovarian contractility in the monkey. Five female rhesus monkeys were treated with natural estrogens, 5 mg/day for three weeks, after which ovarian contractility was studied in vitro in one of the ovaries. Estrogen treatment was followed by progesterone, 25 mg/day for three weeks, after which the contractility of the remaining ovary was similarly measured. Responses to autonomic agents and prostaglandins were studied in both groups. Spontaneous ovarian contractility and ovarian contractile responsiveness to prostaglandins and norepinephrine were found to be enhanced after progesterone treatment. Cholinergic agonists had a stimulatory effect after progesterone and an inhibitory effect after estrogens. Our results suggest that ovarian contractile responsiveness is modified by the local steroid environment, perhaps through intracellular changes in cyclic AMP.


The effect of prostaglandin F2alpha on corpus luteal function in the rhesus monkey. An experimental model was set up to show whether PGF2alpha caused luteolysis when adequate chorionic gonadotropin was administered. Four mature female rhesus monkeys were studied for two cycles each. HCG was given after ovulation until day 36. PGF2alpha was given on days 28 and 29 in one cycle, while the other served as control. In five out of the total eight cycles studied, the luteal function was maintained up to day 36. However, gradual decline of serum progesterone was noted after days 22 to 26 despite the continuous administration of hCG. Administration of PGF2alpha on days 28 and 29 did not accelerate CL regression. It was also noted that hCG in doses of 500 IU did not maintain CL function in three of four cycles when surgical stress was added.


Neutrophils and the mechanism of IUD action in rats. Current research on the mechanism of action of the IUD has focused on a local, low-grade endometritis in preventing blastocystic implantation. With rabbit antisera to rat neutrophils, a neutropenic state was induced in rats having a silk suture in one uterine horn. Assay for nidation sites in the severely polymorphdepleted rats revealed no implantation sites in the IUD horn and an average of 5.8 sites in the control horn, suggesting that inflammation plays a relatively minor role, if any, in the mechanism of action of the IUD.


Action of cyproterone acetate on the accessory organ of reproduction in prepubertal and sexually mature rats. The effects of cyproterone acetate on the accessory organs of immature, prepubertal, and sexually mature adult rats were studied. Administration of CA to immature 30-day-old rats for 15 days markedly decreased the silaic acid in the caput and cauda epididymides and decreased the secretory activity of the accessory glands. Treatment of adult rats with CA for 15 days caused an increased incorporation of 3H-uridine into RNA and of 3H-phenylalanine into protein in the caput and cauda epididymides. The percentage of increase in the specific activity of RNA and protein was higher in the cauda epididymidis than in the caput epididymidis.


Oxidative and glycolytic metabolism of semen components by washed guinea pig spermatozoa. The concentration of several potentially metabolizable substances in guinea pig semen and the ability of these substances to support ATP synthesis and the motility of guinea pig sperm have been determined. Both glucose and fructose were present in high concentration in semen and were equipotent at the concentration tested in maintaining high levels of ATP and a high rate of motility. Lactic and pyruvic acids also supported a high rate of sperm motility but maintained lower levels of ATP. These constituents of guinea pig semen, as well as the metabolites alpha-glycerophosphate, succinic acid, and beta-hydroxybutyric acid, are oxidized at unusually high rates. The active oxidative metabolism of guinea pig sperm is compared with that of human sperm which is primarily glycolytic.


Stimulation of ejaculated human spermatozoa by caffeine. Caffeine (1, 3, 7-trimethyl-2, 6-dioxypurine), a cyclic nucleotide phosphodiesterase inhibitor, increased the motility of ejaculated human spermatozoa. Both the percent motility and the grade of forward progression were significantly increased (P smaller than 0.001) and maintained in the presence of 6 mM of caffeine for up to five hours at 37 C. Increases in percent motility greater than two-fold occurred among the groups having low initial motility. Increases in samples with a high initial percent motility (greater than 60%) were not as dramatic, but they were still significant. In addition, the grade of motility or forward progression was increased in all groups up to one full unit. The longevity of the spermatozoa, judged by activity, was also increased for up to five hours.


Effect of centrifugation and seminal plasma on motility and fertility of stallion and bull spermatozoa. The effect of centrifugation of diluted and undiluted semen on equine and bovine spermatozoan motility and fertility was examined, as was the effect of seminal plasma and dilution on stallion spermatozoa during incubation before and after freezing. Centrifugation at 370 g or 829 g was not detrimental (P greater than 0.05) to prefreeze or postfreeze motility if a final concentration of 10% seminal plasma was present. A reduction of seminal plasma from 10% to 2% significantly (P smaller than 0.05) reduced motility. A centrifugal force of 956 g significantly reduced prefreeze but not postfreeze motility of spermatozoa in undiluted semen, regardless of seminal plasma concentration. With a dried skim milk extender, prefreeze and postfreeze motility was greater in samples containing 20% seminal plasma. Motility was depressed by high and low concentrations of seminal plasma. The fertility of frozen or unfrozen stallion spermatozoa was not depressed (P greater than 0.05) by centrifugation at 310 g for 3.5 minutes. In contrast, the fertility of bull semen was significantly (P smaller than 0.05) lowered by centrifugation at 270 g for three minutes. Further, the fertility of centrifuged, diluted bovine semen was lower (P smaller than 0.05) than centrifuged, undiluted semen.


Spontaneous reanastomosis of the vas deferens after surgery: a case report. Spontaneous reanastomosis of the vas on one side in a 40-year-old man was caused by a sperm granuloma. Leaking spermatozoa bridged the gap between the cut ends; this was followed by epithelial canalization. From review of the literature it was concluded that the best preventive measure is light fulguration of the cut ends and then their separation by covering one cut end by its faschial sheath.


Tubal ligation and pregnancy: mechanism of recanalization after tubal ligation. The mechanism of recanalization after resection of a segment of the fallopian tube by the Pomeroy sterilization method is discussed. At the level of resection, the epithelial lining of the fallopian tube tends to regenerate, covering the split ends and planes of cleavage of the resected surfaces and forming slitlike spaces and blind pouches lined by tall columnar cells. Scarring and subsequent retraction of both severed ends of the tubes tend to result in approximation; in some cases, the resected ends and the epithelial lining bridge the gap between the lumina, re-establishing patency of the tube. Surgical procedures that prevent approximation of the resected ends of the fallopian tubes or methods that seal the lumina should produce failure rates lower than those obtained with the classic Pomeroy sterilization method.


Effects of postovulatory estradiol benzoate administration on women's ovarian function. The effects of postovulatory estradiol benzoate (EB) administration were studied in six women having normal biphasic, presumably ovulatory, menstrual cycles. EB was administered intramuscularly at dose levels of 0.5 and 10 mg daily for 5 consecutive days. The study covered a total of 30 cycles; parameters studied were basal body temperature, urinary excretion of estrogens and pregnanediol, and characteristics of cervical mucus. EB had no effect on the length of the luteal phase nor on urinary pregnanediol excretion, but a dose of 10 mg/day caused a significant prolongation of the next follicular phase. The elevated estrogen excretion observed at the end of the cycle supports the hypothesis that gestagenic deprivation causes menstrual bleeding.


Quantitative analysis of immunoglobulins and albumin in secretion of female reproductive tract. Total protein, IgG, secretory IgA (SIgA), IgA, IgM, and albumin were quantitatively analyzed in 115 cervical fluid specimens from healthy, adult women. Although albumin was the most predominant protein among those that were analyzed (17.2% of total protein), IgG was the major immunoglobulin in this secretion (8.7%). A highly significant correlation between the levels of IgG and albumin and a mean IgG-albumin ratio similar to that of serum suggest that both proteins originate from the circulation. Although the main type of IgA was of the secretory type (4.4%), serum type IgA (smaller than 1.0%) was close to 2. The mean levels of IgG and albumin (but not of SIgA) of the postpartum group were significantly higher than those of the normal subjects. The number of children, the current method of contraception, and present and past local infections had no effect on the immunoglobulins and albumin in this secretion when the mean levels of these components were analyzed and compared with the "normal" values.


Transfer of uterine implantation blastocysts to the oviduct in mice. Although ectopic pregnancy is a common occurrence in humans, it occurs only rarely in other species. In the mouse, confinement of the blastocyst to the oviduct fails to result in implantation at that level. In prior studies, these confined blastocysts had not been exposed to the uterine environment. We transferred blastocysts from the uterus, at a time when implantation would normally occur, to ligated oviducts of pseudopregnant mice. Although enlargement of the blastocysts, loss of the zona pellucida, and adherence to the tubal epithelium were noted, implantation did not occur. The contributions to the embryo by the uterine environment were not sufficient to allow implantation within the oviduct.


Infertility practice and Orthodox Jewish law. The evaluation and management of infertility problems in Orthodox Jewish couples can be made difficult because of restrictions which may be imposed by religious rules. The origins of the religious laws governing problems such as semen collection, diagnostic and therapeutic genital surgery, menstruation, and homologous and donor insemination are reviewed. Contemporary Rabbinic authorities are quoted; their opinions may serve as guidelines for the patient and physician dealing with infertility problems.


[Extinction of brain activation responses to direct electrical stimulation of its structures in normal awake cats]. In unrestrained cats, repeated electric stimulation of the mesencephalic reticular formation (MRF), center median (CM) of the thalamus, and different cortical areas: both the low--and the high--threshold points (in regard to the brain activation), with the threshold strength current evoked similar EEG reactions of activation which diminished and disappeared after 3--5 repetitions of the stimuli.  The moderate strength current evoked, apart from the EEG activation, pseudoviolent movements (turning of the head, etc.) and changes in the breathing rate.  All these reactions could be extinguidhed by sufficient number of repetitions of stimuli, the effector reactions disappearing first, the EEG changes--last.  The essential difference of the stimulation effects emerged when the strong current stimulation was used.  In this case, when stimulating the high-threshold cortical points, the EEG and effector reactions could be abolished during long enough repetition of the stimuli, but it was impossible when stimulating the low-threshold cortical points, the MRF or CM: all the reactions stayed intense and stable, the animals became highly irritated.  The data obtained are discussed from the point of view of the authors' concept of the interaction between the activating and integrative analysing mechanisms of the brain.


[Mixed and antidromal cortical responses evoked by electrical stimulation of the splenial portion of the corpus callosum]. In anesthetized cats, antidromic and orthodromic components were shown to be represented unequally in the mixed responses to the splenium stimulation recorded from various areas of the cortex.  Orthodromic component makes about 50 per cent of the mixed response amplitude in the area 17, while in the area 18 and in middle part of the suprasylvian gyrus it makes about 70 per cent.  In the regions beyond focus the mixed responses disappeared because of falling out of the orthodromic impulses.  During antidromic recording of the responses, only deep layers got activated at weak stimuli, while both deep and superficial ones were involved at stronger stimulation.


[Seasonal changes in the influence of prostaglandin E2 on corticosteroid biosynthesis by rabbit adrenals]. The biosynthesis of corticoids from exogenic tritated progesteron with and without addition of progstaglandin E2 in incubation medium was studied in rabbits in spring and in summer.  C-14 inclusion into aldosteron, cortisone, and 11-dehydrocorticosterone in the spring rabbits was considerably higher than in the summer ones.  Prostaglandin E2 suppressed the biosynthesis of the final fraction of corticosteroids in the spring rabbits and did not change the C-14 inclusion into corticosteroids in summer.


[The influence of prolonged painful stimulation on adrenaline and noradrenaline concentration in different regions of the brain and in the adrenals of white rats before and after removal of the superior cervical sympathetic ganglia]. Prior to removal of the sympathetic ganglia, the sustained nociceptive stimulation decreased the adrenalin and noradrenalin content in the medulla oblongata, cerebellum, and the midbrain while increasing it in the adrenal glands.  After the removal, the stimulation was followed by a shorter (except in the medulla oblongata) decrease of the adrenalin and noradrenalin content in the same brain areas; in the adrenal glands the noradrenalin contents increased.


[Functional features of the electroreceptors of several electric and non-electric fish]. The effect of the electric, magnetic, and mechanical stimulation of the electroreceptors was studied in marine skates (Trigon pastinaca, Raja clavata, and Torpedo marmorata) by recording single nerve fiber responses.  The following two types of backbround dischanges were found with regular activity and bursting.  In some fibres the bursts of impulses appeared simultaneously with respiration while in the others the activity was suppressed.  The current threshold of the clear-cut responses were equal to 10- minus 9--10- minus 11 A/mm-2.  The activity of the electroredeptors was modified by introducing a conductive or nonconductive object into the water.  High sensistivity of the electroreceptors to weak magnetic field was shown.  The biological role of the electroreceptors is discussed.


[A mathematical model of cardiac rhythm disorders in the presence of rapid auricular electrical activity]. The model is based on regularities of changes of the refractory period in the atrioventricular conductive system as well as changes of the delay of impulse conduction in this system during coupled electric stimulation of atria.  With the aid of the model, the conditions of accurring heart-rate disturbances were considered (increase in the delay of the atrio-ventricular conduction,  Wenkenbach's periodicity, fallin out of two, three or more successive excitations of the ventricles, etc.) on fast electric activity of the atria.  Quantitative correlations were established between the parameters characterizing changes of the refractory period and the period of atrias excitations which produces a ventricles rhythm disturbance.


[The mechanism of action of parenterally administered protein hydrolysates on the exocrine activity of the pancreas]. In dogs with fistulae of the stomach and the pancreatic duct as well as in dogs with artificially made partition between the stomach and the duodenum, effect of protein hydrolysates administered into the blood, on the pancreas secretion, was shown to manifest itself only in conditions of unopposed transition of the stomach acid contents into the duodenum.  In the mechanism of protein hydrolysates action on the pancreas secretion, the major part is played by their ability to stimulate the stomach secretion and the transition of the acid content into the duodenum.  Under the influence of the acid contents, release of the secretin occurs in the duodenum which induces the secretion of the pancreatic juice.


[Renal and extrarenal mechanisms of potassium homeostasis following a potassium load]. In rats, per os i.v. administration of 1.25 per cent KCl solution (840 mcEq/100 g.b.w.) increased concentration of potassium in the blood and tissues, as well as the urine excretion of potassium.  The excretion rate depended on the amount of potassium load.  After adrenalectomy and aldactone injection, the potassium excretion decreased.  This, apparently, indicates that the adrenocortical steroids exert an influence on the transport of potassium.  Hence, both the renal and extrarenal systems play an important role in the potassium homeostasis.


[The effect of cold acclimatization on thermoregulation in the Mongolian gerbil, Meriones unguicilatus]. The long-term (45 days) acclimation of the Mongolian gerbil, in contrast to albino rats, is not accompanied by adaptive changes in general metabolism, chemical thermoregulation, bioelectric skeletal muscles activity, and tissue respiration in muscles and liver.  The effect of short-term intermittent cooling in the Mongolian gerbil is of short duration and does not manifest itself in increased stability of body temperature.  The calorigenic effect of norepinephrine may be only observed after a long-term adaptation.  The pattern of adaptation to cold in the Mongolian gerbil, a typical habitant of continental climate of Central Asia, depends on the stability of its normal thermoregulation.


Effects of cortisol on uterine eosinophilia and other oestrogenic responses. The effects of cortisol on several oestrogenic responses of the rat uterus were measured. Whether injected i.v., simultaneously with oestradiol-17 beta, or i.p,, 12 h before the oestradiol, cortisol had no effects on the oestrogen-induced increases in uterine glycogen, protein and DNA contents. In contrast, cortisol inhibited both uterine eosinophilia and the increase of wet weight. Both responses show the same higher sensitivity to i.v. injection than to i.p. injection of cortisol. Inhibition of both responses by cortisol follows identical dose-response curves. These data support our hypothesis that the water-imbibition effect of oestrogen i- mediated by uterine eosinophilia and is thus related to the eosinophil receptor system.


Sertoli cell origin of testicular androgen-binding protein (ABP). In this report it is suggested that the specific adrogen-binding protein (ABP), previously shown to originate in the testis of rat and other species, is produced by the Sertoli cells. This suggestion is based upon the following experimental findings: 1) ABP was found in high concentrations in testicular efferent duct fluid but only in trace amounts in inter-tubular lymph. i) ABP could be recovered from crude preparations of testis tubules, but not from Leydig cells from the same testes. 3) Testes whose germinal epithelium had been severly damaged by gamma irradiation showed no decrease in ABP content. The transport of ABP to epididymis was also preserved as judged from the levels of ABP in caput epididymis. 4) Testes that were completely devoid of germ cells following prenatal gamma irradiation showed high levels of ABP, These high levels approached zero following hypophysectomy, but could be restored by FSH administration to the hypophysectomized animals. ABP has been well characterized and now provides a valuable experimental tool as an indicator of Sertoli cell function.


Single and mixed infections of avian infectious bronchitis virus and Mycoplasma gallisepticum. Comparative studies of the clinical signs, pathological changes, multiplication of the pathogens and serological responses were made of groups of chickens infected with either or both avian infectious bronchitis virus and Mycoplasma gallisepticum. Electron microscopic examination of thin sections of infected tracheas showed that the multiplication of M. gallisepticum was greatly enhanced in the tracheas of chickens which had been previously or simultaneously infected with avian infectious bronchitis virus. The enhanced multiplication of mycoplasmas resulted in more severe clinical signs, pathological effects and serological responses. These findings confirm the general belief that avian infectious bronchitis infection may precipitate latent mycoplasma infection. Similar experiments were made in the allantoic sacs of 10-day embryonated hens' eggs. The results again show that there was at least a one hundred-fold increase in the multiplication of M. gallisepticum in eggs simultaneously or previously infected with avian infectious bronchitis virus. These findings emphasize the importance of using mycoplasma-free eggs for the production of virus vaccines.


The protection of ferrets against influenza by immunization with a split influenza vaccine. Ferrets were vaccinated with either A/England/42/72 zonal purified influenza queous vaccine containing whole virus particles, or a subunit vaccine containing the purified hemagglutinin and neura-minidase proteins of A/England/42/72 adsorbed onto aluminium hydroxide. The immune status of the ferrets was assessed by reaction to challenge with virulent live virus of the homologous strain. In the preliminary experiment reported, the subunit vaccine conferred greater protection than the zonal purified vaccine.


Neuraminidase assay of influenza vaccines. At present influenza vaccines are standardized on their haemagglutinin content only. Recently it has been shown that both neuraminidase and haemagglutinin antibodies are important in providing protection against the influenza virus. We have, therefore, developed an automated neuraminidase assay, based on the enzymic method of Kendal, but modified to minimise the interference by sucrose. A neuraminidase standard has been established and samples assayed against the standard have a coefficient of variation of plus or minus 6.8 percent. The assay has been adapted to measure the neuraminidase antibody level in serum. The neuraminidase to haemagglutinin ratio has been determined for various influenza strains. Both neuraminidase and haemagglutinin titres are being compared with results obtained by the single radial diffusion method.


Influenza virus and ciliary beating of the respiratory epithelium in sensitized animals. Normally, under good survival conditions, the ciliary beating can be observed and recorded during hours after dissection of the respiratory epithelium. When laboratory animals have been immunized locally against influenza virus (under delayed type hypersensitivity conditions) the ciliary beating stops within 3 to 6 minutes when the same virus is put in contact with the sensitized mucosa. The specificity and the immunological conditions of this stopping have been analyed and the practical importance of this new test in cellular immunology is discussed.


Delayed hypersensitivity to influenza virus in man. Delayed hypersensitivity to influenza antigen has been demonstrated in man by means of skin tests and 'in vitro' lymphocyte transformation. Adjuvanted influenza vaccine enhances this response, in contrast to non-adjuvanted vaccine in commercial use.


'Non-specific' mediators in host defense against respiratory viruses: assay methods and results. Understandably, there is interest in further defining the nature of 'non-specific' defenses. This, in fact, would clarify the specific nature of their appearance and action, as well as their role in host defense. Therefore, 'non-specific' defenses in my discussion will be taken to mean non-immune defenses in virus infections. Tissue or respiratory tract organ cultures are convenient means of studying host defenses in the absence of immune responses. Factors to be controlled in making observations with such systems will be discussed which help 'in vitro' results to parallel 'in vivo' findings. 'In vitro' and 'in vivo' methods and results probing importance of mucous barrier, ciliary activity, temperature and pH will be reviewed. Critical points in separating cellular and humoral immune functions from non-specific mediators will also be touched on. This distinction is complicated by the fact that both limbs of immunity work in close concert with macrophages, PMN's complement and other inflammatory responses. Interferon and macrophage mobilization are examples of 'non-specific' responses which have more recently become complicated by their recognition as part of the effector limb of the cell-mediated immune responses. Finally, methods will also be presented for distinguishing human interferon of the immune-specific variety associated with cell-mediated immunity from that produced by viral infection of non-immune cells.


The immune cellular response tested by lymphocyte transformation in the streptococcal infections. Thirty-five children between 6 and 17 years treated in the clinic for scarlet fever, rheumatic fever and other non-streptococcal infections as controls, were tested by lymphocyte transformation to four streptococcal antigens. In all cases of scarlet fever and especially of rheumatic fever the lymphocytes were better stimulated by streptococcal products than in the control group. The SO and the MAP fraction showed a good stimulating activity. The response in the rheumatic fever patients was not influenced by the steroid treatment, nor by the stage of the illness. A parallelism with high humoral and cellular responses to SO at the beginning of the acute rheumatic fever was observed, followed by a dissociation of both responses during the evolution with the maintenance of the cellular one and the decrease of the ASO titre.


Duration of circulating and secretory antibody and cell-mediated immunity following immunization. An important consideration in evaluating vaccines is the duration of immunity. The only really important measure of this immunity is the protection against infections and/or illness at various time intervals, following natural or artificial challenge. There are few data of this sort, more commonly immunity is estimated by measuring serum antibody, in many instances an erroneous measure. Serum antibody levels to respiratory viruses fall only slightly 6 months following infection or immunization. It is difficult to assess the duration of antibody for much longer than this, because of problems with intercurrent infection. With respiratory bacterial infections, e.g. pneumococcal pneumonia, parenterally-induced immunity probably lasts for only several months. Secretory antibody induced by inactivated viral vaccines, seems to persist for about a year, after having reached a peak level at about 4-6 weeks following immunization. Work with the live attenuated polio virus vaccine indicates longer lasting immunity, with detectable antibody persisting for up to 34 months. Restimulation with the inactivated polio virus vaccine produced no evidence of a secondary response (memory). Following booster immunization with influenza very little evidence of memory is seen. Cell-mediated immunity (CMI): in guinea pigs BCG sensitization can be demonstrated for at least 2-9 months. In humans, intracutaneous BCG immunization leads to positive tuberculin reaction in 6-10 weeks, and skin sensitivity lasts an average of about 4 years. There is contradicting data as to the duration of protection against infection following BCG immunization. Local and systemic CMI have been shown to exist independently of each other in experimental animals and man.


Assessment of immunity to influenza using artifical challenge of normal volunteers with influenza virus. The candidate humoral mediators of protection against influenza include antibody (Ab) to the viral hemagglutinin (H) or neuraminidase (N) in serum or respiratory secretions. In the present studies these mediators were evaluated principally by low dose live virus challenge of previously vaccinated volunteers. Following IM vaccination with H3N2 virus, Ab appeared in both serum and secretions and direct quantitative relationship between the two was noted. Among individuals vaccinated intranasally with comparable doses, the serum and secretion Ab responses were similar to those after IM vaccination. Irrespective of immunization method, there was a better correlation between protection and titers of serum Ab than titers of Ab in nasal secretions. After vaccination with an N-specific vaccine, an inverse correlation between titer of serum anti-neuraminidase Ab and quantity of virus in secretions occurred. This was reflected in occurrence of illness in those with low Ab titers, infection only in those with intermediate Av titers and no evidence of infection in those with high Av titers. In a naturally occurring outbreak with the England variant, a similar pattern of infection responses occurred among persons with varying titers of serum anti-hemagglutinin Ab. This suggests that, in man, Ab to the H and to the N results in similar host responses to infection and that the extent of infection is determined by magnitude ofAb present.


Assessment of resistance to influenza virus infection in animal models. The antibody response and immunity to challenge infection were determined in ferrets immunized with inactivated influenza vaccine in saline or adjuvant. Adjuvanated vaccines induced variable titres of serum antibody, and the degree of immunity to challenge infection was directly related to the titre of serum HI antibody induced by these vaccines. Conventional doses of saline vaccine did not induce serum HI antibody, and the ferrets were completely susceptible to challenge infection. Infection with live virus produced a more solid immunity to challenge infection than immunization with a adjuvant vaccines, even though immunization induced higher titres of serum HI antibody. Ferrets previously infected with a heterotypic influenza A virus, but not other viruses, produced serum HI antibody in response to subsequent immunization with inactivated influenza vaccine. Similar results were obtained in hamsters and mice. Thus, the failure of animals to produce antibody in response to immunization with saline inactivated vaccines was due to the absence of a previous priming infection; this prior experience would be a feature of most volunteers. Live virus infection produced nasal antibody in ferrets, but inactivated vaccines only induced serum antibody. This may explain the more solid immunity observed following infection; however, at the time of challenge infection, no nasal wash antibody could be detected. Immunization with inactivated vaccine in Freund's complete adjuvant and influenza virus infection both produced a cell-mediated immune response; thus, the difference in the degree of immunity induced by these two immunization procedures are probably not due to differences in the cell-mediated immune response. However, cell-mediated immunity was measured by skin tests and by macrophage migration inhibition tests with spleen cells; the reaction of cells from the respiratory tract may be more important, but was not measured in these studies.


Effect of neuraminidase on potency of inactivated influenza virus vaccines in mice. The protective effect of neuraminidase was studied in a mouse protection test using isolated neuraminidase of A2/Aichi/68(H3N2) virus and the complete recombinant virus A/eq(Heq-1)-HK(N2) as antigens. Immunized mice were protected against A2/Aichi(H3N2) challenge virus; however, the protection rate was low in comparison to animals immunized with comparable amounts of the complete A2/68(H3N2) virus9 Furthermore there was no cross-protection against A2/Asia/57(H2N2) challenge virus. The protective effect of neuraminidase was not impaired by Tween-ether treatment of the A/eq(Heq-1)-HK(N2) recombinant virus.


Contribution to the standardization of influenza neuraminidase inhibition tests, based on enzyme- antienzyme kinetic studies. 1. From enzyme-antienzyme kinetic studies a method for the determination of anti-n2-neuraminidase antibodies has been developed. This procedure is applicable in routine diagnosis. 2. Considering the present conditions, the reversibility of the virus neuraminidase antigen-antineuraminidase antibody complex has been proved. The inhibition must be described as a non-competitive one. 3. As a reference value for comparative immunity estimations the relative antibody concentration T150 is defined as the point of reciprocal serum dilution, where under a given substrate concentration the inhibited enzyme activity vi is half of the non-inhibited enzyme activity vo. 4. For the calculation of T150 a simple formula is derived from the Michaelis-Menten kinetic. Under present conditions the value of T150 is in a range of 1.5 log 2 dilution steps plus or minus the specific serum dilution T150 uninfluenced by possible variable parameters such as enzyme activity, substrate concentration and inhibitor concentration.


Potency of influenza vaccines: mouse protection experiments in correlation to field studies in man. The seroconversion rates have been studied following vaccination of human volunteers with two commercial influenza vaccines. Vaccine A did not give a significant increase of hemagglutination-inhibition titers. Vaccine B, on the other hand, raised the titers 2- to 8- fold, depending on the pretiters of the individuals. The potency of the same vaccines has been tested using mouse protection experiments: vaccine B gave significantly better protection rates, as measured by survival as well as by reduction of lung lesions. These results give additional evidence that the use of mouse protection experiments for the evaluation of different influenza vaccines is meaningful.


Immune response to combined live influenza virus vaccines administered intranasally. The immune responses in volunteers vaccinated intranasally with a live influenza virus vaccine containing attenuated A and B strains have been studied. The hemagglutination-inhibiting antibody titers for both components were determined a few weeks after vaccination. They demonstrated that a good antibody response had been induced in a high percentage of volunteers. The hemagglutination inhibiting titers and percentage of seroconversion for both components were comparable to the results obtained after the intranasal administration of each component separately.


An 'in vitro' marker of attenuation for live influenza virus vaccine candidates. Organ cultures of ferret tracheal rings maintained in tubes for up to two months supported the growth of several strains of influenza A viruses produced by genetic recombination. These strains were developed as possible candidates for live attenuated influenza virus vaccines. By observing the effects on cilia and titrating the growth in eggs it was possible to rank these various influenza strains in order of descending virulence for the respiratory epithelium in organ culture. This rank corresponded in general to the virulence of these viruses for human volunteers. Viruses too virulent to use as vaccine strains in man regularly destroyed the ciliated epithelium of ferret trachea while strains attenuated for man usually did not. Although the ciliated respiratory epithelium of ferret was not as sensitive as that derived from human embryonic trachea, this test did identify all the tested strains too virulent for evaluation in human volunteers.


Immunization schedules for influenza.

Immunization schedules for influenza. Although on an individual basis and for some selected closed groups immunization against influenza has proved its value over the last 30 years, it has not resulted in prevention of epidemics. The impact of vaccination on national morbidity and mortality statistics has been disappointing. The reasons for this apparent failure of disease control by immunization are discussed. It is concluded that influenza vaccination has not been practised on a large enough scale to achieve an obvious effect on the spread of the viruses in open communities. The groups of people who deserve special attention in order to minimize the damages caused by influenza epidemics are considered. Advantages and disadvantages of available types of vaccine, methods and routes of administration are assessed. It is suggested that systematic application of present knowledge would probably have more impact in the near future than efforts to increase vaccine efficacy. The main current deficiencies are identified as the difficulty of producing adequate supplies of properly constituted vaccines at short notice and the general reluctance to accept annual large-scale immunization programmes. In the view of the authors these logistic and administrative problems could be resolved by long-term coordinated planning between relevant authorities and vaccine manufacturers.


Reactions to injected influenza vaccine. Almost 6,000 healthy adults inoculated with influenza vaccine were asked to record daily for 5 days the presence or absence of defined local and general symptoms; 70 percent completed and returned the record. The overall incidence of subjective reactions derived in this way was high; approximately 50 percent complained of local pain and 40 percent of general symptoms. Only about one-third of vaccinees recorded no symptoms. The incidence of subjective local reactions was about 8 percent higher in women than men. Among 5,600 vaccinees a total of 63 days sickness absence was attributed to reaction in the week of immunization (1.1 days per 100 employees). Almost all the absences were of less than 3 days duration, unsupported by a doctor's certificate. In the week of vaccination sickness absence among vaccinees was not higher than that of non-vaccinees. Although the incidence of subjective reaction was found to be high and vaccination might itself cause some sickness absence, most of the reactions appear to be trivial. Among 700 employees who were interviewed 1 or 2 days after vaccination in 1973, the incidence of what was judged to be significant local pain and erythema was about 5 percent. Nevertheless,the reactions appear to have an important effect on acceptance of influenza vaccination. From the answers given to a questionnaire 30 percent of employees who refused vaccination did so either because they previously had symptoms after vaccination, or because others had told them of their occurrence. In approximately 16,500 injections, only 2 patients had an acute reaction resembling anaphylaxis.


Group A streptococcal M protein vaccine: protection following immunization via the respiratory tract. Previous studies have shown the efficacy of parenteral immunization of volunteers with purified type 1 M protein against challenge with homologous streptococcui (J. clin, Invest. 52: 1885, 1973). A double-blind study was conducted on 21 adults immunized by aerosol-spray into the nasopharynx, and 23 controls who received saline placebo. Two booster doses were given at monthly intervals and approximately 30 days later vaccinees and controls were challenged with homologous streptococci (10-6/ml) by swabbing the pharyngeal-tonsillar areas. Throat cultures, leukocyte counts, temperatures and physical signs and symptoms were followed to assess infection. Illness was defined as a positive throat culture, oral temperature of larger than or equal to 38 degrees C, a WBC count of twice baseline or greater than 10,000 per mm-3, exudative pharyngitis and adenopathy. Of the 43 subjects, 13 were ill by all criteria (10 controls, 3 vaccinees [p smaller than .02]); 21 were well by all criteria (6 controls and 15 vaccinees); and 10 exhibited some but not all positive criteria (7 controls, 3 vaccinees). Positive throat cultures following challenge were obtained in 19 controls and 5 vaccinees (p smaller than 0.001). There was no correlation between the pre-challenge serum antibody titer and the development of subsequent illness. It is concluded that local topical immunization with a M protein vaccine offers significant type-specific protection against challenge with streptococci.


The anti-MAP and anti-group A carbohydrate antibodies response in streptococcal human infections. The streptococcal infection cases from two outbreaks were serologically examined against two components of the streptococcal cellular wall; the M associated protein (MAP), by a latex agglutination test and the group A carbohydrate (A-CHO), by passive hemagglutination technique. Many positive cases with high levels of both antibodies were found, in rheumatic fever and glomerulo-nephritis, comparatively with the acute streptococcal infections. The differences were statistically significant. The results were correlated with the ASO titres and with the dermic cellular response.


Newcastle disease.

Newcastle disease. Newcastle Disease of chickens is of interest in comparative pathology because the causal virus has a wide range of pathogenicity and is of only one antigenic type. The virus can be grown to high titre and is easily titrated. Both live attenuated and inactivated vaccines give effective protection under experimental conditions. Vaccination of commercial chickens is associated with variable results, and it is now possible to investigate the factors which complicate vaccination. These include the presence of interfering respiratory infections, immunosuppressive disease of viral origin, variations in the techniques of vaccination and in the programmes of revaccination. Challenge with lethal virus given by aerosol is severe and allows an accurate assessment of the relationship between serum HI levels and the degree of protection. In the later stages of the immune process most but not all protection is associated with the IgG fraction of serum. There is also an increasing amount of evidence to show that local immunity in the respiratory tract is important in the early stages of the immune process.


Newcastle disease: effects of vaccines and vaccination techniques of hemagglutination-inhibition (HI) antibodies and clinical resistance. Different vaccines (the lentogenic Hitchner-, the avirulent Ulster- and the mesogenic Beaudette strain) and different vaccination schemes (spray, dip, drinking water) were used in 1380 broiler-type chicks on the first day of life and at four weeks. In all these experiments an immunity status was induced sufficient to resist to a strong individual challenge infection, two weeks after the second vaccination. This uniform resistance showed no correlation with the very heterogeneous HI titers which were obtained two weeks after the second vaccination. The titers were, however, strongly dependent both of the vaccine strain used and on the vaccination method. The Beaudette spray-vaccinations provoked the highest titers. The highest post-challenge titers were seen after vaccinations with the lentogenic and avirulent strains and the lowest after-challenge titers were seen when the mesogenic Beaudette strain was used. Birds with high HI titers showed a marked decrease of these HI titers two weeks after challenge, while there was an increase of HI titers in birds with low post-vaccinal titers. It may be concluded that clinical resistance to NCD infection is not directly correlated with high-vaccinal HI titers. Experimental infection by natural route is essential to judge the immune status of fowls.


Immunity to mycoplasma infections of the respiratory system in the domestic fowl and turkey. The mycoplasma infecting and sometimes causing disease of the respiratory system in both species are M. gallisepticum and M. synoviae, while M. meleagridis affects the turkey only. M. gallisepticum has received the greatest attention and natural infection is reported to result in protection varying greatly in degree and duration. Vaccination with living organisms either by the respiratory or intravenous routes results in a high degree of protection against air sac challenge for at least a few weeks. Killed organisms administered intravenously result in a lesser degree of protection to this challenge but when given intranasally provide no protection. M. gallisepticum survives in the respiratory tract and perhaps in other organs and tissues for long periods after infection, perhaps in latent form. However, intercurrent infection and other avian respiratory pathogens may precipitate multiplication of the organism and disease in birds apparently resistant to challenge with mycoplasma alone. The mechanism of protection has not been extensively studied and production and significance of local antibody or a cell-mediated response have not been determined. However, bursectomized chickens vaccinated against M. gallisepticum show much less resistance compared with controls but the exact part played by humoral antibody is still in doubt. Similar observations have been made with M. synoviae.


Efficacy of inactivated Mycoplasma pneumoniae vaccine in man. Inactivated Mycoplasma pneumoniae vaccine produced antibody responses that reached protective levels in most individuals. These were almost equivalent to natural infection in amount and duration, and they persisted more than a year. Adverse effects were inconsequential. At Keesler Air Force Base, Mississippi, where the annual rate of mycoplasma pneumonia varied from 20 to 50 per 1000 since 1959, protection studies were conducted in 21, 199 men in 1964-1966 and in 13, 892 men in 1969-1971. This produced a 36 percent reduction in bronchitis and a 45 percent reduction in pneumonia due to all etiologies in the frist study and 37 percent and 48 percent respectively in the second study. The protective efficacy of the second vaccine was 87 percent for acutebronchitis and 66 percent for pneumonia due to Mycoplasma pneumoniae. Persons who developed Mycoplasma pneumoniae infections in spite of vaccination did not experience more severe illnesses.


Prevalence of concurrent diabetes mellitus and idiopathic facial paralysis (Bell's palsy). Diabetes mellitus was present in 11.4 per cent of 684 patients with Bell's palsy, in 28.4 per cent of the sixty-seven with recurrent or bilateral palsy, and in 16.8 percent of the 440 with palsy who were thirty years or older.  Diabetes was present in only 3.8 per cent of 27,399 persons thirty years or older who had never had Bell's palsy and who underwent multiphasic health testing.  These figures clearly indicate that diabetes is more common among patients with Bell's palsy than among persons who have never had that disease; and that the risk of Bell's palsy is increased in patients with diabetes.  The diabetic patient is more prone than the non-diabetic person to nerve degeneration, and this tendency to nerve degeneration is not age-related.  Although 10 per cent of our patients with Bell's palsy and known diabetes were younger than thirty-nine years, we now advise screening for blood sugar elevation only for patients who are forty years or older, or who have recurrent or bilateral facial paralysis.


Prevalence of diabetes and glucose intolerance in 199 offspring of thirty-seven conjugal diabetic parents. We have studied the prevalence of diabetes and glucose intolerance, both cross-sectionally and longitudinally, in a cohort of 199 offspring of conjugal diabetic parents.  Although the prevalence of already known diabetes was low (11.5 per cent), twenty-eight of 123 tested offspring (23 per cent), ranging in age from ten to sixty (mean 32.6) years had latent diabetes on their initial glucose tolerance test.  Eighty of 123 tested offspring had a normal initial glucose tolerance test.  Forty-one of those whose first glucose tolerance test was normal were retested after a mean of 9.4 years and showed no significant change in mean glucose tolerance.  On the basis of questionnaire data only, we estimate that cumulatively 36.5 per cent of offspring will have diabetes by the age of sixty years.  However, if all offspring are routinely surveyed with glucose tolerance tests, 60 per cent will have abnormal glucose tolerance by the age of sixty years.  In view of the high prevalence of asymptomatic latent diabetes, genetic studies of diabetes should not be based on questionnaire data.  In six families all offspring were diabetic; in twenty-one families there was a mixture of diabetic and nondiabetic offspring; and in ten none of the offspring was diabetic.  The finding of families with no diabetic offspring suggests the possibility of genetic heterogeneity of diabetes in the parents.  Most of the parents had maturity-onset diabetes with a mean age at diagnosis of 54.5 years of age.  Diabetes among their offspring was generally of a mild maturity-onset type.  Only 2 per cent of offspring at risk had developed juvenile-onset type diabetes.  Thus the prevalence of any metabolic abnormalities in offspring of two maturity-onset type parents cannot be assumed to be relevant to the offspring of parents with juvenile-onset type diabetes.


Influence of oral glucose ingestion on splanchnic glucose and gluconeogenic substrate metabolism in man. To evaluate the role of splanchnic and peripheral tissues in the disposal of an oral glucose load, splanchnic exchange of glucose, lactate, pyruvate, glycerol and amino acids was determined in ten healthy subjects in the basal state and for three hours following the oral ingestion of 100 gm. of glucose.  Following glucose ingestion, splanchnic glucose output rose rapidly, reaching values two to three times the basal rate at fifteen minutes and returning to baseline by ninety minutes.  A secondary rise in splanchnic glucose output occurred at 150 minutes and coincided with a secondary increment in arterial glucose.  Total splanchnic glucose output over three hours was 40 plus or minus 3 gm., representing a total increase of only 15 plus or minus 3 gm. above basal splanchnic glucose output.  The peak rise in blood glucose was directly proportional to the increase in splanchnic glucose output.  Arterial concentrations of alanine, lactate and pyruvate rose by 15, 65 and 80 per cent, respectively, following oral glucose.  These arterial elevations were preceded by a 75-100 per cent inhibition of splanchnic uptake of alanine and lactate; in the case of pyruvate there was a reversal from a net uptake in the basal state to a significant net splanchnic output after glucose ingestion.  Arterial glycerol fell by 50 per cent and was accompanied by a comparable fall in splanchnic uptake.  It is concluded that in normal, postabsorptive man, (a) the major portion of a 100 gm. oral glucose load is retained within the splanchnic bed; (b) only 15 per cent of the ingested glucose is available for disposal by peripheral tissues as increased (above-basal) glucose utilization; (c) the height and shape or the oral glucose tolerance curve are largely determined by the rate and pattern of splanchnic glucose escape; (d) glucose-induced hyperlactatemia, hyperpyruvicemia and hyperalaninemia are due at least in part, to altered splanchnic exchange of these substrates.


Modulation of fatty acid metabolism by glucagon in man.  I. Effects in normal subjects. This investigation was designed to examine the contribution of glucagon to the regulation of free fatty acid (FFA) metabolism in man. The acute effects of exogenous glucagon upon the concentration of plasma FFA and its metabolites: acetoacetic acid (AcAc), beta hydroxybutyric acid (BOH), and triglyceride (TG) were assessed in five normal male subjects. The threshold of response was determined by administering intravenous glucagon in a graded dose-response fashion, andassessing the magnitude of change in FFA and its metabolites. A prompt rise in FFA concentration occurred after glucagon injection, and was associated with a synchronous elevation in both AcAc and BOH concentration in plasma. The rise in FFA concentration was maximal with the glucagon dosage of 0.5 mug./kg. with no greater FFA response seen athigher hormone dosage. In contrast, the rise in plasma ketones was linearly related to thedosage of glucagon but not to the corresponding rise in FFA concentration. The behavior of plasma triglyceride concentration following glucagon challenge also contrasted with the linear rise in ketone concentration. At the lowest dosage of glucagon, TG concentration rose transiently, while at the highest two dosages the concentration of TG was reduced. The transitional dosage of hormone at which no change in TG concentration was observed was approximately 0.5 mug./kg. Comparing our in vivo data with the conclusions reached from published liver perfusion studies, it can be suggested that in the dosages employed, glucagon acutely elevates plasma FFA concentration and modulates their metabolism by augmenting conversion to AcAc and BOH while tending to reduce conversion to TG in man.


Barium or gastrografin: which contrast media for diagnosis of esophageal tears? In order to study the effects of the commonly employed contrast media alone and in combination with bacteria in the mediastinum, various mixtures of barium and meglamine diatrizoate (Gastrografin) with and without flora were instilled in the mediastinum of 29 domestic cats. The animals were killed for pathological studies at serial time intervals following mediastina injection. These data demonstrate that water-soluble contrast media cause no significant histological reaction. Barium causes granuloma formation but has no additional deleterious effects when mixed with flora. Barium has superior physical properties of mucosal coating and radiographic density. It should be employed as the contrast agent of choice in difficult clinical problems with regard to esophageal tears. A water-soluble contrast study of the esophagus may be utilized as the initial diagnostic procedure, but if this is normal a follow-up barium esophagram is recommended.


Carcinoma and epithelial dysplasia complicating ulcerative colitis. The pathology of 19 specimens of carcinoma complicating ulcerative colitis, resected in the University Department of Surgery of the General Infirmary at Leeds, was reviewed with particular reference to the incidence of epithelial dysplasia. The carcinomas were found to be more frequently multiple, more evenly distributed in the large bowel, and much more often of atypical macroscopic appearances and of mucoid histological type than ordinary colorectal carcinomas, but the proportion of poorly differentiated tumours complicating ulcerative colitis was not as high as previously reported. Of the patients in our series 26% are alive and well at least 5 years after surgery. Unequivocal epithelial dysplasia was demonstrated in some part of the large intestine in 15 of 19 specimens with colitis carcinoma, but was also found in 4 of 14 specimens from a "control" series of patients with longstanding total colitis but without carcinoma. Clearly, therefore, the finding of dysplasia in a rectal biopsy of a patient with colitis is not a reliable guide to the presence of a frank carcinoma elsewhere in the bowel. Whether it indicates a special predisposition to the development of a growth in the future, as might be postulated from the analogy of similar changes in other organs, cannot be determined on the data of this study. The fact that epithelial dysplasia when present in colitis is often patchy in distribution and frequently spares the rectum even in patients with definite carcinomas makes a negative rectal biopsy particularly unreliable in deciding on the absence of a tumour or the lack of predisposition to it. Multiple biopsies from different parts of the colon as well as the rectum would thus seem to be desirable if mucosal sampling is to be employed as a screening test.


Control of gastric emptying by osmolality of duodenal contents in man. During the steady state perfusion of the duodenum, recovery of a duodenal marker (polyethylene glycol 4000) can be used to quantify intraduodenal volumes. We used this technique to measure and time the recovery from the duodenum of a second nonabsorbable marker, phenol red. When liquid test meals containing phenol red were placed in the stomach, the rate of gastric emptying could be quantified from the appearance of the maker in the duodenum. The osmolality of either test meal or duodenal perfusates was then varied so that the osmotic control of gastric emptying could be examined. Gastric emptying was fastest when duodenal contents were isotonic; nonisotonic duodenal contents slowed emptying whether these conditions were achieved by nonisotonic duodenal perfusates or by emptying of a nonisotonic test meal. However, nonisotonic solutions in the stomach and in the jejunum did not slow gastric emptying, so long as duodenal contents were isotonic. Osomoreceptors, which slow gastric emptying, are present in the duodenum but not in the jejunum or stomach.


Early lymphoepithelial relationships in human appendix. A combined light- and electron-microscopic study. The fine structure of human appendix was studied from the earliest stages of lymphoid development in fetuses to the definitive relationships found in children up to 8 years old. Follicular accumulations of lymphocytes were observed first in the mesenchyme immediately beneath epithelium which contained a predominance of goblet cells on the surface and in the crypts. Larger accumulations of lymphoid cells in older fetuses were intimately related to surface epithelium but not to the epithelium of crypts. At the point of invasion of lymphoid cells into surface epithelium, the goblet cell population diminished and epithelial cells displaying a morphologically distinct form of differentiation were observed. They were characterized by the presence of irregular microvilli or microfolds and numerous apical micropinocytotic vesciles. This follicle-associated epithelium (FAE) appeared ultrastructurally identical with epithelium in chicken bursa of Fabricius, mouse Peyer's patch, and rabbit appendix, which has been shown to be capable of transporting ferritin and India ink tracer from the lumen to underlying tissue. It appeared identical to specialized epithelial cells of adult human Peyer's patches. FAE was maintained through the neonatal period into childhood. We speculate that the biological significance of FAE is to provide a channel through which antigens may stimulate clonal proliferation and seeding of B-lymphocytes throughout the lamina propria of internal mucous surfaces.


Radioimmunoassay for motilin.

Radioimmunoassay for motilin. A specific radioimmunoassay for motilin has been developed with the use of antisera to porcine motilin raised in guinea pigs. Highly purified 125-I-motilin was used as the tracer and the sensitivity range was 10 to 320 pg. No cross-reactivity was demonstrated with gastric inhibitory polypeptide, secretin, glucagon, gastrin, cholecystokinin-pancreozymin, or vasoactive intestinal peptide. In dogs with denervated pouches of the fundus of the stomach and Mann-Bollman fistulae, duodenal alkalinization resulted in an increase in gastric motor activity in the fundic pouch with a corresponding increase in serum motilin.


Inhibition of gastric emptying is a physiological action of cholecystokinin. This study was designed to determine whether cholecystokinin (CCK) plays a physiological role in the inhibition of gastric emptying. Physiological conditions were simulated by giving CCK by continuous intravenous infusion rather than by bolus injection, by using doses known to be distinctly submaximal for pancreatic protein secretion, and for gallbladder contraction, and by releasing endogenous CCK. The rate of gastric emptying was determined in 4 dogs with gastric fistulas by measuring the volume of fluid remaining in the stomach 10 min after instillation of 300 ml of 0.15 M NaCl. Rate of emptying was studied during intravenous infusion of saline (control) and of different doses of 98% pure CCK, commerically available 20% pure CCK, synthetic COOH-terminal octapeptide of CCK (OP-CCK), pentagastrin, and heptadecapeptide gastrin. The effect of endogenously released CCK was studied by measuring the rate of emptying of solutions in which different concentrations of tryptophan replaced equiosmolar amounts of NaCl. The d50's of 20% pure CCK (3 U kg minus-1 hr minus-1) and of OP-CCK (125 ng kg minus-1 hr minus-1) for inhibition of gastric emptying were about the same as their D50's for cholecystokinetic and pancreozyminic actions. By contrast, although both pentagastrin and heptadecapeptide gastrin inhibited gastric emptying, the doses required for this action were much higher than the D50's required for stimulation of gastric acid secretion. The effectiveness of OP-CCK indicates that inhibition of gastric emptying is attributable to CCK itself and not to an impurity in the CCK preparation. We have confirmed this directly by showing that pure CCK is a potent inhibitor of gastric emptying. Tryptophan also inhibited gastric emptying. In other dogs pancreatic protein secretion and gallbladder contraction were shown to be stimulated during the time tryptophan was inhibiting gastric emptying. This evidence supports the view that inhibition of gastric emptying is one of the physiological actions of CCK, but in the case of gastrin it must be regarded as a pharmacological action.


Thiamine transport across the rat intestine. I. Normal characteristics. The characteristics of normal thiamine transport across the intestine were studied in rats using intact intestinal loops and everted jejunal segments. In vivo studies with [35-S]-thiamine hydrochloride revealed, in all segments of small intestine, saturation kinetics for low thiamine concentrations (0.06 to 1.5 muM), but a linear relationship between high concentrations (2 to 560 muM) and absorption. Moreover, in vitro studies of net transmural flux using everted jejunal sacs demonstrated movement of [14-C]-thiamine hydrochloride against a concentration gradient only when low, but not when high, thiamine concentration was used, so that the serosal to mucosal ratio became significantly greater than the initial value of one. Pyrithiamine, 2 muM, dinitrophenol, 200 muM, norethylmaleimide, 100 muM, and ouabain, 10 muM, reduced the net transmural flux of 0.2 muM thiamine. In contrast, these inhibitors had no effect on 20 muM thiamine. When unidirectional flux across the jejunum was measured, saturation kinetics was again demonstrated for low thiamine concentrations. This phenomenon, however, was abolished by the addition of pyrithiamine, which exerted competitive inhibition on thiamine absorption. Anoxia and sodium lack reduced intestinal uptake of 0.5 muM thiamine to 58% and 74% of normal, respectively, but did not affect uptake of 50 muM thiamine. Lowering the marked with low thiamine concentrations (O10, 1.648) than with high concentration (Q10, 1.127). Stirring of the water layer reduced Km to 59% of unstirred value, while Vmax and permeability coefficient remained unchanged. Finally, movement of low concentration thiamine against an electrical gradient was observed under conditions of electrical short circuiting and zero potential difference. In contrast, no such effect was seen with high concentrations. These studies suggest that there exists in the rat a dual system of intestinal thiamine transport. At low concentrations, thiamine is absorbed by an active process; at high concentrations, transport across the intestine is largely a passive movement.


Effects of bombesin and bombesin-like peptides on gastrointestinal myoelectric activity. Gastrointestinal myoelectric activity was studied during intravenous infusion of bombesin and bombesin-like peptides in conscious dogs with electrodes chronically implanted at different levels between the stomach and the rectum. The peptides used were bombesin, the COOH-terminal hepta- octa-, and nonapeptide of bombesin, and litorin, a new natural peptide isolated from the skin of Litoria aurea. Bombesin significantly increased the frquency of pacesetter potentials (PP) in tha antrum, duodenum, jejunum, and ileum. In the duodenum and jejunum the increase of PP frequency showed linear correlation  with the reduction of PP amplitude. The propagation velocity of PP was clearly reduced. Spikes were not affected in the antrum and ileum, whereas they were abolished in the duodenum and jejunum. In the duodenum the increase of PP frequency and the slowing down of propagation velocity was followed by the loss of PP phase lock and the appearance of a characteristic electric pattern, consisting of an irregular sequence of small and slow potentials ("electric disorganization"). The mechanical counterpart was the disappearance of intraluminal pressure activity. In the colon the effect of bombesin on electric activity was not consistent. Neither the COOH-terminal heptapeptide nor the octapeptide of bombesin showed a significant effect on myoelectric activity, whereas the effect of COOH-terminal-nonapeptide and litorin was similar to that of bombesin. Thus, the characteristic electric changes of PP produced by bombesin appear to be related to the sequence of the nine amino acids in the COOH-terminal residue of the bombesin molecule.


Morphological and functional effects of bile salts on rat colon. By correlating morphological observations with quantitative measurements of net water transport, we determined whether bile salts altered colonic absorptive cells. Epithelial alteration was equivocal and water absorption was uninhibited during infusions of 1 mM deoxycholate or of concentrations of taurocholate less than 10 mM. In contrast, 3 mM deoxycholate and greater than 10 mM taurocholate caused severe altertion of colonic epithelium and inhibited water absorption. These studies suggest that bile salts in the low concentrations normally found within the colon have little effect on colonic structure or water absorption. On the other hand, abnormally high concentrations of bile salts do alter colonic mucosal structure and function.


Value of gallbladder B-scan ultrasonography. The gallbladder B-scans of 20 patients who had subsequent surgery were separated into three categories based upon certain sonographic criteria. Our data, in this limited series, revealed gallbladder pathology in each patient who had any one or combination of the following scan characteristics: (1) internal echos, (2) irregular wall, or (3) absence of recognizable gallbladder sonolucency. The category which demonstrated a normal sonographic gallbladder, namely a smooth wall and no internal echos, contained a number of false negatives which proved to have either small stone cholelithiasis or extraphepatic ductal obstruction. Within the described limitations, the B-scan can be a valuable test in confirming the significance of a radiographically nonvisualized gallbladder or in detecting a biliary tract lesion in a patient with a disease entity that precludes radiographic visualization by conventional techniques.


Serum free proline and free hydroxyproline in patients with chronic liver disease. Serum free proline and free hydroxyproline were determined in 71 patients with liver disease and in 62 control subjects. The group with liver disease included 60 patients with liver cirrhosis and 11 with chronic active liver disease. Forty-five of the cirrhotic patients were alcoholics, 9 of which were studied during an episode of alcoholic hepatitis. The control group consisted of 24 healthy volunteers, 19 primary malnourished patients, and 19 severely ill patients without liver disease. The values obtained in normal subjects were quite constant; no differences related to sex or age were detected. In patients with malnutrition, and especially in severely ill patients, the proline values were always below the normal limit. Patients with nonalcoholic cirrhosis or chronic active liver disease had serum proline and hydroxyproline values similar to those of normal subjects. However, the patients with alcoholic liver cirrhosis had proline and hydroxyproline values significantly higher than the normal group. Furthermore, in patients with alcoholic hepatitis the serum free proline values were significantly higher than in the other groups. The results suggest that alcohol might have a direct effect on proline metabolism or facilitate its release from the liver cell.


Noncirrhotic presinusoidal portal hypertension associated with chronic arsenical intoxication. A 39-year-old male with bleeding esophageal varices due to portal hypertension was observed. The patient had taken an arsenical preparation during a period of 12 yr because of psoriasis and subsequently developed keratotic changes of the palms and soles of his feet and an epithelioma of the scrotum. Physical examination was unremarkable except for splenomegaly and skin lesions. Liver function tests were normal; a needle biopsy of the liver (right lobe) showed nonspecific changes. Combined hepatic and umbilicoportal catheterization revealed, on splenography and portography, huge esophageal varices and patent portal vein; dilation, distortion, and cut-off of many intrahepatic portal branches were found. A marked gradient existed between the free portal venous pressure (25 mm Hg) and the wedged hepatic venous pressure (9.5 mm Hg). Hepatic blood flow, portal PO2, cardiac output, cardiac index, and blOOD volume were within normal range. Arteriographies did not reveal arteriovenous shunts in the splanchnic or splenic vessels. A splenorenal shunt were performed and a wedged biopsy of the liver (left lobe) revealed nonspecific changes. Three years later the patient had not experienced any episode of hemorrhage or hepatic encephalopathy but developed an epithelioma of the tongue. No known cause could be incriminated in the pathogenesis of the portal hypertension. However, there was unequivocal chronic arsenic intoxication. Toxic hepatitis, cirrhosis, noncirrhotic portal hypertension, and hemangiosarcoma of the liver have been reported with the intake of arsenicals. Thus, it is suggested that in this patient, presinusoidal portal hypertension was secondary to chronic arsenical intake associated with marked intrahepatic vascular changes seen on portography.


Jaundice due to erythromycin estolate.

Jaundice due to erythromycin estolate. A 43-year-old male developed abdominal pain and jaundice after the administration of erythromycin estolate. The diagnosis was strongly suspected on clinical grounds, but ultimate confirmation depends upon the demonstration of biochemical and morphological alterations after challenge with the drug.


Small intestinal mucosal injury in the experimental blind loop syndrome. Light- and electron-microscopic and histochemical studies. Microscopic (light and electron) and histochemical abnormalities have been demonstrated in the jejunum of rats with the blind loop syndrome. Three groups of animals were studied: normal control animals, and animals with either self-filling (SF) or self-emptying (SE) blind loops. Vitamin B12 malabsorption and bacterial overgrowth occurred only in those animals with SF blind loops. Three jejunal segments were studied: the blind loop segment and the jejunal segments proximal and distal to the blind loop. In the animals with the blind loop syndrome, those with SF blind loops, the most striking findings occurred in the blind loop itself, with similar but less marked changes in the jejunum distal but not proximal to the blind loop segment. Hypertrophy of both crypts and villi was evident with focal abnormalities of villus architecture. Approximately 10 to 20% of the columnar cells in the upper half of the villi were swollen and vesiculated. By electron microscopy microvilli demonstrated a variety of degeneration changes and the glycocalyx and terminal web were disrupted. Mitochondria and endoplasmic reticulum (ER), both smooth and rough, were swollen. Concentric whorls of parallel membranes and long, curvilinear rough ER were present in the cytoplasm. Histochemically, there was loss of enzymatic activity in the epithelial brush border, mitochondria and ER. Inasmuch as bacterial invasion of the jejunal mucosa was not seen, the etiology of these changes is not known but may involve bacterial "toxins" or products of bacterial metabolism. These morphological observations demonstrate that both brush border and intracellular injury occur in the jejunal epithelial cell of rats with the experimental blind loop syndrome.


[Balanced synthetic diet in gynaecological operations]. A balanced synthetic diet, the so called astronaut-diet was given to nine gynaecological patients with recto-vaginal fistulas or operation for aplasia of the vagina. By the reduction of stool quanitity and the change to an almost sterile stool, wound healing was very much enhanced. This diet is recommended for some operations involving the vagina and rectum.


[Basic pathology of sigma and rectum in geriatric-gynaecological laparotomies]. This is a report on 49 women with sigmoiditis and 66 women with sigmoido-rectal carcinoma. These cases were collected in 18 women hospitals in West-Berlin over a period of 10 years, from 1960 until 1969. All patients were at least 60 years old. In most of these cases laparotomy was performed for gynaecological reasons. The post-operative mortality was 40.9%. It is pointed out that the whole spectrum of diagnostic procedures was not fully utilized; today laparoscopies and ultrasound are added when indicated. Inflammatory processes of the sigma as well as sigmoido-rectal carcinoma are typical geriatrical diseases that will increase with rising life expectancy thus constituting an important part of the differential-diagnostic section of geriatric gynaecology.


[Psychological problems in heterologous insemination]. Heterologous insemination is being established in increasing numbers in Anglo-Saxon, middle and north European countries, although some medical, legal and theological-ethical aspects are still not settled. Our experiences with this type of sterilisation in regard to marriage stability and psycho-physical development of the children are positive. During the months of insemination, even with psychically balanced couples, conflicts arise such as disturbed vita sexualis (sexual life) emotional stress situations or aggravation from the husband's inferiority complex which can endanger the success of the treatment or even the marriage. The doctor in charge of insemination has the responsibility to discuss and analyse arising problems with the couple so that both husband and wife are emotionally and intellectually prepared when pregnancy occurs.


[The serum and uterine muscle concentrations of Cephradin and Cephalothin]. On the day of operation 2 grams of Cephradin was given intravenously to 33 patient and 2 grams of Cephalothin was given intravenously to 31 patients. At various time intervals, uterine tissue and serum was removed. The mean serum concentrations of Cephradin were 3 times higher and the mean tissue concentrations of Cephradin were 7-8 times higher than the corresponding concentrations of Cephalothin. Because of the difference in protein binding of the examined cephalosporins (Cephradin 6%, Cephalothin 60-65%) the difference for the antimicrobially active portion (Protein free portion) is even more favorable for Cephradin. The significance of these pharmacokinetic differences for the treatment are obvious.


[Morphological study on 167 Wertheim-operation specimens (author's transl)]. The squamous cell carcinoma of the uterine cervix was examined in 167 cases with the regard to the spreading and the dependence of metastazation on the size of the primary tumor. Up to a primary tumor size of 5 + 10 + 10 mm a metastazation of 1,58% in the lymph nodes, of 30,9% at stage Ib and of 57,6% at stage II was found. The metastazation rate from one stage to the other seems not to be continuously but skippy. The hysterectomy seems to be an entirely sufficient therapy in case of a microcarcinoma, provided that accurate three-dimensional measurement proves that the size does not exceed 5 + 10 + 10 mm. The Wertheim's operation should give its best results at stage Ib, as long as it includes the careful removal of the pelvic lymph nodes.


[Long term results after Latzko operation for vesico-vaginal fistula]. The Latzko operation for the closure of a vesico-vaginal fistula is still not well enough known. Urologists have voiced doubts about the functional quality of the late results after this operation. Pouches in the bladder wall which could lead to infection and stone formation are said to occur frequently. Follow-up studies regarding this question are not known. Therefore the patients treated at the University Hospitals Erlangen-Nuernberg and Graz between 1962 and 1972 were subjected to a standardized follow-up protocol. Of 37 women who had the Latzko operation, 24 women returned for a follow-up examination. 19 women had no intermittent urinary tract disease and the follow-up results were normal. In many instances a scar corresponding to the previous fistula could not be detected on cystoscopy. Immediately after the operation, one patient had a recurrent fistula and has not been re-operated. Four women had significant urinary tract findings or had intermittent urinary tract disease. These complications were possibly due to the operation in one case and likely due to the operation in a second case. Our findings suggests that the Latzko operation is the method of choice for the treatment of the typical post-hysterectomy vesico-vaginal fistula.


[On the question of recurrent carcinoma in-situ of the uterine cervix]. Among 280 patients with carcinoma in-situ of the uterine cervix treated from 1961 to 1970, a review of the histology revealed 50 patients in which the carcinoma in-situ had been removed with certainty by conization. None of these patients was found to have had a recurrence. In the literature, 13 observations of a recurrence of carcinoma in-situ after complete removal by conization were located among 3, 129 cases. It is concluded that true recurrence after complete removal of carcinoma in-situ by conization is rare and that conization is adequate treatment for carcinoma in-situ for patients who attend regular follow-up examinations.


[Pull through operation for endometriosis of the recto-sigmoid]. Case report on endometriosis of the recto-sigmoid which caused intestinal stenosis. Concomitant endometriosis of the right ovary was removed by a right salpingo-oophorectomy. The recto-sigmoid endometriosis was resected and intestinal continuity established by the pull through method of Hollenbach preserving the anal sphincter. The patient was well 6 years following the operation.


[Aspiration biopsy in ovarian tumours].

[Aspiration biopsy in ovarian tumours]. The value of fine-needle aspiration biopsy in ovarian tumours is determined comparing cytological and histological findings. In our material of 53 cases cytological diagnostic accuracy is 92.6%. Further cytological classification occasionally is possible. Additional studies are necessary to permit final conclusions about the applicability and diagnostic accuracy of the method.


[Unexpected situations during laparatomia]. This is a report of a case of a malignant Haemangiopericytom in a 49 year old woman. Four solitary tumors were discovered in connective tissue of the right side of the pelvis, which histologically presented a variety of forms, partially mature, partially immature embryonal tissue, in which the atypical proliferating vessel-portion was predominant.


Fitness of karyotypes in Drosophila pseudoobscura. In the dynamics of the survival of chromosomal polymorphism selection may be operating at the genic level, at the chromosomal level or at the supergene level. Tests designed to distinguish between these levels were run on Drosophila pseudoobscura. There was no evidence for heterosis, a necessary requirement for gene-determined chromosomal polymorphism A strong chromosmal selection was observed. No evidence was found for the presence within one locality of more than a single superallele for each supergene (equals gene order). These results are compared to those found by others.


The response of dopa decarboxylase activity to variations in gene dosage in Drosophila: a possible location of the structural gene. A location of the structural gene(s) for dopa decarboxylase (EC 4.1.1.26) is proposed on the basis of enzyme determinations in a set of duplication-bearing aneuploids, which revealed only one dosage-sensitive region in the Drosophila genome. This region lies between 36EF and 37D on the left arm of chromosome 2.


Further evidence for selective differences between isoalleles in Drosophila. A number of separate strains of Drosophila pseudoobscura were inbred for 38 generations of brother-sister mating with forced heterozygosity for two alleles of either the octanol dehydrogenase or esterase-5 locus. Crosses were set up within each of these inbred lines such that simple mendelian ratios were expected, and eggs from these crosses were placed on media with additions simple chemicals likely to interact with alleles of the two loci--octanol and ethanol for the ODH locus and tributyrin and triacetin for the E-5 locus. Similar crosses were set up involving parental flies with normally heterozygous genetic background as a control.--Significant deviations from mendelian expectation were observed in inbred E-5 flies grown on tributyrin, inbred ODH males grown on octanol, and inbred ODH females grown on ethanol. There was also a strong effect of octanol medium on males of one of the inbred E-5 lines, and a weak effect of tributyrin medium on ODH inbred females.--The probability that these results reflect interactions between these loci and the environment is assessed in the light of differences between the present results and those obtained at earlier stages of inbreeding.


Evidence for the single phase pairing theory of meiosis. The segregation pattern of an attached X chromosome with several Y-autosome translocations conflicts with the expectations based on the distributive pairing hypothesis because the chromosomes segregating from the translocation configuration include both exchange and non-exchange chromosomes. The results of the second experiment involving three compound chromosomes go even further; they suggest that the essential association which determines the segregation of nonhomologous elements is in fact set up prior to the time of crossing over.


Chemical selection of mutants that affect alcohol dehydrogenase in Drosophila. II. Use of 1-pentyne-3-ol. We describe a procedure for the selection of alcohol dehyrogenase negative mutants in Drosophila. The method consists of exposing eggs and larvae to low concentrations of 1-pentyne-3-ol dissolved in the culture medium. Only those flies with greatly reduced levels of alcohol dehydrogenase activity survive. In addition, genotypically negative flies die if their mothers are alcohol dehydrogenase positive. Using this procedure and formaldehyde to generate mutants, we were able to detect seven alcohol dehydrogenase negative mutants out of 350,000 individuals subjected to selection. At least five of the mutants contain small deletions that include the alcohol dehydrogenase locus.


Fitness of allozyme variants in Drosophila pseudoobscura. I. Selection at the PGM-1 and Me-2 loci. We have studied in Drosophila pseudoobscura the effect of allozyme variation on seven fitness components: female fecundity, egg hatchability, egg-to-adult survival under near-optimal and under competitive conditions, rate of development under near-optimal and under competitive conditions, and mating capacity of males. Three genotypes at each of two loci, Pgm-1 and Me-2, have been studied in various combinations. These two loci are highly polymorphic in natural populations of D. pseudoobscura. Statistically significant differences involving one or more genotypes exist for all components of fitness. No single genotype is best for all fitness components; rather the relative fitnesses of genotypes are reversed when different parameters are considered, or when they are studied in different environmental conditions. Also, the average egg-to-adult survival and rate of development are better when different genotypes are reared together than when they occur in pure culture. Four different modes of selection have been uncovered by our experiments. These forms of selection may account for the persistence of the two allozyme polymorphisms in nature, and for previously observed seasonal fluctuations of the allelic frequencies in natural populations.


Genetic organization and adaptive response of allozymes to ecological variables in Fundulus heteroclitus. Populations of Fundulus heteroclitus, (Cyprinodontidae) a widespread coastal marine fish, were studied in control and artificially heated environments on the North Shore of Long Island, New York to determine (1) patterns of variation in biochemical phenotypes and (2) the extent to which this variation reflected adaptation to environmental characteristics. Variation at three of twelve polymorphic isoenzyme loci from the warm water population was beyond the range of variation among control populations, and resembled those determined for populations living at more southern latitudes. Hence, these differences were interpreted as adaptations to warm environments. Significant differences in allele frequencies and zygotic proportions at ten of twelve isoenzyme loci were found associated with differences in environments, sexes, and/or age classes. These data strongly support the view that protein polymorphisms are adaptive. Several observations suggested that selection acts upon multilocus phenotypes rather than upon those of single loci. Several di-locus phenotypic distributions were demonstrated to be nonrandom, and those that exhibited similar patterns of dependence over years were postulated to be maintained by selection. Highly heterozygous fish exhibited superior viability when cohorts were compared over successive years. The consequences of the polygynous mating system in this species for maintaining genetic variation and for allowing rapid evolutionary response to a variable environment are discussed.


Studies of mutations in T4 control genes 33 and 55. Available mutations in transcriptional control genes 33 and 55 of coliphage T4 have been examined. By complementation analysis and map position, 15 mutants (13 in T4D, 2 in T4B) have been shown to lie in gene 33 and 6 (5 in T4D, 1 in T4B) in gene 55. According to patterns of suppression and recombination, these mutants define three distinct amber sites in gene 33 and also three distinct amber sites in gene 55. All of these mutations are true amber mutations, in apparent contrast to some traditional T4 "amber" mutants which grow in su+ E. coli CR63 but not in su minus E. coli B because of a strain difference other than the su+ determinant. Evidence is presented that, contrary to previous suggestions (BOLLE et al. 1968; pulitzer and geiduschek 1970), the gene 33 product is absolutely essential for T4 development.


A genetic and biochemical study of histidine biosynthesis  in Micrococcus luteus. Histidine auxotrophs of Micrococcus luteus strain ATCC 27141 were induced by treatment of the parent strain with N-methyl-N'-nitro-N-nitrosoguanidine. Auxotrophs were biochemically characterized by examining culture accumulations of histidine intermediates, using paper chromatography and the Bratton-Marshall test, and growth responses to L-histidinol. his(IG) mutants failed to accumulate Pauly-positive imidazoles; his(EAHF) mutants accumulated 5-amino-1-ribosyn-4-imidazole carboxamide; hisB mutants accumulated imidazoleglycerol; hisC mutants accumulated imidazoleacetol; hisD mutants, but did stimulate all other histidine mutants, blocked at earlier steps in the biosynthetic pathway. In addition, imidazoleglycerol phosphate dehydrase activity was assayed in representative mutants of each class. hisB mutants lacked activity for this enzyme.--Two -point, three-point, and cotransformation analyses resolved linkage relationships of histidine genes and in two gene clusters aided in determining their sequences. Histidine biosynthetic genes exist in at least four separate, unlinked regions of the chromosome. One histidine gene cluster linked to a tryptophan gene cluster and appears to be contiguous in the sequence his(IG) -his(EAHF)-trpE-trpC-trpA. A second and unlinked histidine cluster has the tentative gene sequence his(EAHF)-hisB-hisC-his(EAHF). The hisD gene and an unclassified mutant site his-94 are not linked to any of the other histidine genes examined in this study or to each other.


Clonal age and the proportion of defective progeny after autogamy in Patamecium aurelia. The relation of mortality and the proportion of progeny with reduced fission after autogamy to the clonal age in Paramecium aurelia was investigated. This relation is not linear but the proportion of defective progeny in creases stepwise. The observations are in agreement with those expected from the calculations of the number of deleterious mutations in the micronucleus.


Expression in organ culture of agouti locus genes of the mouse. CLEFFMANN (1963) reported that pigment cells of the lethal yellow (A-y/a) genotype change to black immediately upon cultivation, and continue to produce black pigment unless sulfhydryl reagents are added to the culture medium. We have attempted to repeat this observation and have not been able to do so. We also have been unable to indue the synthesis of yellow pigment by adding glutathione to cultures of agouti (A/A) skin. We therefore suggest that hypotheses which attempt to explain the action of agouti locus genes on the basis of effects of sulfhydryl compounds be considered with caution.


Genetic polymorphism and evolution in parthenogenetic animals. II. Diploid and polyploid Solenobia triquetrella (Lepidoptera: Psychidae). Genic polymorphism at sixteen enzyme loci of four different chromosomal races of Solenobia triquetrella (bisexual, two diploid parthenogenetic races and tetraploid parthenogentiic) has been studied by starch gel electrophoresis. Isolated small diploid bisexual populations have rather uniform allele frequencies at all loci which we have studied. Diploid and tetraploid parthenogenetic individuals of this species are in general as heterozygous as bisexual ones. All parthenogenetic local populations are different from each other in the Alps. These parthenogenetic genotypes cannot be derived from a common ancestor through single mutations but rather bear evidence for a polyphyletic origin of parthenogenesis in Solenobia triquetrella. In the marginal distribution areas of the species in northern Europe single genotypes are spread over far larger areas than in the mountain regions of central Europe. This may be due to the old origin of parthenogenesis and polyploidy in northern Europe. No new parthenogenetic and polyploid strains have lately arisen in the regions outside of the Alps.


Natural selection and age-structured populations. This paper studies the properties of a new class of demographic parameters for age-structured populations and analyzes the effect of natural selection on these parameters. Two new demographic variables are introduced: the entropy of a population and the reproductive potential. The entropy of a population measures the variability of the contribution of the different age classes to the stationary population. The reproductive potential measures the mean of the contribution of the different age classes to the Malthusian parameter. The Malthusian parameter is precisely the difference between the entropy and the reproductive potential. The effect of these demographic variables on changes in gene frequency is discussed. The concept of entropy of a genotype is introduced and it is shown that in a random mating population in Hardy-Weinberg equilibrium and under slow selection, the rate of change of entropy is equal to the genetic variance in entropy minus the covariance in entropy and reproductive potential. This result is an information theoretic analog of Fisher's fundamental theorem of natural selection.


The deterministic behavior of self-incompatibility alleles. For a system of n self-incompatibility alleles, neglecting mutation and random drift, it is shown that the completely symmetric equilibrium is locally stable, and any allelic frequency less than q equals 1 + a minus the square root of 1 + a-2, where a equals [2(n minus 1)]- minus 1, will increase. For all n, q greater than (2n)- minus 1, but if n greater than 1, q is approximately equal to (2n)- minus 1.


Unilateral clubbing of the fingernails in patients with hemiplegia. A survey based on an analysis of photographs compared the angle at the base of the fingernail in paralysed hands with controls. The significance of the changes in the nail bed angle in relation to clubbing and atrophy of the finger is discussed.


The general practitioner and the hospital. An experiment in in-patient care. A 3-year rotating programme for three general practitioners as clinica assistants involved in in-patient care is described in the three associated departments of medicine, geriatric medicine and psychiatry in a district general hospital. The appointments were for three sessions a week each. The effectiveness of such rotating appointments is analysed including both the service contribution to the hospital and the educational content from the general practioner's point of view and also the way in which such appointments fit in with the timetable of work in general practice. Such a rotating scheme is most successful in geriatric medicine and psychiatry. Involvement in in-patient care in general medicine proved more difficult. Future schemes might include only 6 months in general medicine, together with 6 months in another specialty.