Behavioral observations of mentally impaired aged: those who decline and those who do not. Behavioral observations were conducted on aged mentally impaired females in an institutional environment on a weekly basis for a 2-year period. An eight-category taxonomy was used to rate the complexity level of the subjects' behaviors. Although the average weekly performance showed significant linear decline over the 2 years, half the subjects remained stable in the level of their behavioral functioning. In comparison to subjects exhibiting declines in level of functioning, the stable subjects evidenced less serious medical conditions, had better comprehension of their situations, were more socially reactive, manifested fewer neurotic conditions, and had greater control over impulses and aggression at the beginning of the project.


Variables associated with the winter camping location of elderly recreational vehical owners in southwestern Arizona. During the winter of 1973-1974, 580 elderly recreational vehicle campers were interviewed in six private trailer parks and four public camping areas in southwestern Arizona and Southern California. The ten sites represented four different types of camping environment: (1) highly developed, relatively expensive urban private trailer parks, (2) moderately developed, relatively inexpensive, rural private trailer parks, (3) authorized, developed, free federal campgrounds, and (4) unauthorized, undeveloped, free public land camping areas. Analysis of variance was utilized to identify those respondent characteristics most strongly associated with the type of site selected for winter residence. The most important of these variables were per person annual income, woman's age, man's education, woman's total preretirement adult camping experience, value of mobile quarters and tow vehicle, and percentage of the year lived in a recreational vehicle. Several financial indicators and certain noneconomic characteristics of females were strongly associated with winter camping location.


Mortality rates among relocated extended-care patients. Eighty-one male extended-care patients relocated due to planned demolition of the building in which their ward was located were followed for 21 mo. from the date of transfer. No increase in mortality rate was detected during the first 3 mo. post-relocation, an interval usually associated with high mortality in elderly persons involuntarily relocated. During the first year after relocation, the death rate was 33.33%, compared to an average annual death rate of 41.20% during the 5 years preceding the move. At 21 mo., half the relocated population were alive. These data contrast with previous studies, most of which show increased mortality after relocation, and lead us to concur with those who suggest that the degree of environmental change involved may be a key factor in explaining variance in relocation effects.


A mathematical model of aging processes. IV. A multivariate analysis of blood pressure in a Montreal and a Utah population. Two populations, one Montreal- and one Utah-based, were studied with respect to heart disease risk factors on a cross-sectional basis. The Utah population afforded consistently lower mean blood pressures than the Montreal population, although there was not evidence that the Utah population was less obese, or had a lower pulse rate. Also, in the Utah population, it was found that the proportion of persons with a family history of heart disease did not differ significantly in the hyper- and normo-tensive groups. Fourteen parameters were investigated in the Montreal population, and the analyses indicated that, when other variables are controlled, age, pulse rate, some measure of serum lipid levels, and a family history of heart disease generally assist in the discrimination between the hyper- and normo-tensive groups, but the obesity measurement did not. In that sense, obesity, on its own, may not be considered a risk factor for hypertension.


Superoxide dismutase in the rat and mouse as a function of age and longevity. Superoxide dismutase was assayed in extracts of a variety of tissues in relatively short-lived (A/J) and long-lived (LP/J) male mice. At 9 mo of age the brains and lungs of LP/J mice contained more of this activity than did those of A/J mice. No significant differences were seen in the other tissues investigated. It was also found that specific activities of total superoxide dismutase in extracts of the brain and liver of male CD Sprague-Dawley rats did not diminish during aging; however, the cyanide-insensitive superoxide dismutase, which reflects the mitochondrially localized enzyme and which constitute only a fraction of total activity, did diminish somewhat with age in liver, but not in brain.


The effects of improved oral hygiene on taste perception and nutrition of the elderly. An experimental group of 12 elderly institutionalized subjects received professional oral hygiene therapy three times weekly for 5 weeks while a control group of 11 received only placebo treatment. The experimental group responded with significantly improved ability to detect sweet and salty tastes but not sour or bitter tastes. No change in taste perception was observed in the controls. Initially, nutrient intake of both groups was suboptimal, especially for folacin. After the treatment period, the intake of calories, hematocrits, hemoglobins, and plasma ascorbic acid concentrations of both groups were satisfactory; plasma proteins were slightly low; plasma folates were at deficiency levels. No change in these blood measurements occurred in either group. It is concluded that improved oral hygiene can enhance taste acuity but that taste perception is only one factor determining nutrient intake.


Verbal-pictorial recoding in the elderly. Older individuals have been reported to use imagery mediation less often than the young in remembering verbal material. To determine whether this is due to a decrease with age in the speed with which verbal stimuli are recoded into pictorial representations, the reaction time of 12 old (63-78) and 12 young (17-25) subjects for matching verbal description to geometric shapes was measured. While the elderly were generally slower, they were identical to the young in being faster at determining whether or not two simultaneously presented shapes were the same than they were a printed description of a shape and a shape. Also, in both age groups, when 1 or more sec elapsed between the presentation of the description and that of the shape, matching times were as quick as when both stimuli were shapes, suggesting that, for the old as well as the young, 1 sec was sufficient time to recode the description into a pictorial form.


The effect of practice on age differences in central perceptual processing. The effects of practice on performance in a dichoptic masking paradigm was assessed across two age groups (mean age - 66.5 and 18.8 years). For both age groups practice resulted in a reliable reduction in the time separation between the onset of the target stimulus and the onset of the masking stimulus required to escape masking. The magnitude of reduction was equivalent in both age groups, indicating that the amount of practice in this study does not ameliorate previously demonstrated age differences in speed of central perceptual processing. On the other hand, the equivalent improvement in both age groups suggests no age-related decrement in the adaptability of central perceptual processes.


Satisfaction with life among persons sixty-five years and older. A causal model. A fully recursive path model heuristically depicting a causative chain between financial situation, health satisfaction, satisfaction with standard of living, satisfaction with family life, and satisfaction with life as a whole was examined. The model was evaluated via path analysis, with parallel analyses conducted for each sex. The data used in this analysis were collected from a national probability sample of persons aged 18 and over living in noninstitutional dwelling units. This present study includes only those persons 65 years and older (N = 301). The model was shown to be an effective predictor of satisfaction with life for each sex. Satisfaction with family was found to make the greatest single impact on life satisfaction. Moreover, the model was determined to be generally consistent throughout.


Retired academics and research activity. A limited literature on the retirement of academics suggests that they do not follow the expected pattern of retirement but may be similar to scientists in retirement. Data collected from 207 academics who retired from state and small private colleges and ivy league and large state universities indicate that to varying degrees they continue to engage in professional activities after retirement. Academics who are high on professional visibility appear to be especially likely to continue to engage in research and/or writing. The importance of psychological centrality of pre-retirement work to post-retirement activity is suggested.


Age profiles of membership in sixteen types of voluntary associations. Several trends are noted which may lead future cohorts of older persons to belong to voluntary associations to a greater extent than current cohorts. Because the best previously available data which could be used to monitor changing membership levels are not two decades old, this study presents detailed age profiles of membership in 16 types of associations. Based on data from the 1974 and 1975 NORC General Social Surveys of the adult population of the USA (N = 2,974), various patterns of age differences in belonging to associations are identified and discussed for both the total sample and by sex. A rank order of the types of associations to which older persons are most likely to belong in 1974-1975 is compared with a similar rank order based on a 1955 survey. The paper concludes by specifying additional research questions which replicate studies should address.


A rapid, simple and sensitive method for the demonstration of central catecholamine-containing neurons and axons by glyoxylic acid-induced fluorescence. II. A detailed description of methodology. The glyoxylic acid-induced fluorescence method for localization of brain catecholamine neurons has been modified. Fluorescence is developed rapidly in cryostat sections of brains fixed by perfusion with 0.5% depolymerized paraformaldehyde and 2.0% glyoxylic acid. Since neither freeze drying nor vibratome sectioning is required, total processing time can be less than 1 hr. Both perikarya and fine varicose axons of norepinephrine- and dopamine-containing neurons can be seen throughout the neuroaxis. The modified technique retains good cytologic integrity and may provide a useful alternative for methods combining histochemical approaches.


Spontaneous seizures: a new mutation in Syrian golden hamsters. Hereditary spontaneous seizures in the Syrian hamsters are described. The seizures occur in 30 to 60-day-old animals from about 2 to 5 hours. The condition is a simple recessive trait, and the gene symbol sz is proposed.


The distribution of 14c from [U-14c]glucose in mice using whole-body autoradiography. Tissue distribution of radioactive carbon from [U-14C]glucose in the mouse in vivo was studied by whole-body autoradiography. The mice were frozen with Dry-Ice-acetone at 0.5, 1, 5 and 30 min, 1 and 24 hr and 1 and 3 weeks after intraperitoneal injection of [U-14C]glucose. Whole-sagittal sections of the frozen mouse, obtained by using a microtome in a cryostat, were dried in a cryostat and autoradiographed. The resulting dry autoradiographs are called untreated autoradiographs in the present work. The sections were then fixed in cold 6% (w/v) HClO4, dried at room temperature and again autoradiographed. Autoradiographs that have undergone this process are referred to as treated autoradiographs. In both untreated and treated autoradiographs, within 1 min following injection of the labeled glucose, the abdominal cavity had the highest autoradiographic density. At 1 hr, density became highest in Harder's, sublingual and duodenal glands, large intestinal mucosa and tongue, and after 3 weeks, no autoradiographic denisty was present.


Palp-antenna, a homeotic mutant in Aedes aegypti. Palp-antenna is a homeotic mutant of Aedes aegypti that modifies the apex of the maxillary palps of both sexes into a variable number of antenna-like segments. The fine structure of the antenna, including sexual dimorphism, is apparent in the mutant palps. Palp-antenna is a sex-linked recessive. a linkage distance of 27.9 +/- 0.45 crossover units from sex is estimated from F2 and backcross data. Penetrance is complete; expressively is variable. The fitness of the mutant compares favorably with that of the wild type. The mapping of the ppa locus lengthens the known chromosome 1 linkage map of Aedes aegypti.


Inherited ateliotic dwarfism in mice. Characteristics of the mutation, little, on chromosome 6. A new autosomal recessive mutation in the mouse, little (lit), has been shown to be located on Chromosome 6. The mutation in the homozygous state causes ateliotic dwarfism that is first detected at 15 days of age by decreased body weight. Long bone lengths are significantly reduced. Skull width, however, is not affected. Female little mice are fully fertile; they may lose their first litters. Although most of the little males sire one or two litters, they rarely sire a third litter. Analysis of pituitary extracts electrophoresed on acrylamide gels reveal a significant reduction of the two anterior pituitary hormones, GH and PRL, in both male and female little mice. Because the little mouse shares a number of similarities with the human ateliotic dwarfism, isolated growth hormones deficiency type I, it may be a useful animal model for this inherited human growth disorder.


Changes in C3 metabolism during protozoan infection (Babesia rodhaini) in rats. Metabolism of the third component of complement (C3) and IgG was measured in rats before and during infection with the hemosporidium agent Babesia rodhaina. In the course of infection, hypocomplementemia and immune complex nephritis developed. During babesial infection, in most animals the half-life of C3 fell sharply, as did serum levels of C3; the catabolic rate for C3 sharply increased, whereas the synthetic rate sharply decreased. In contrast, the catabolic rate for IgG remained unchanged. The alteration in the metabolism of C3 in the face of nonparallel changes in IgG metabolism suggests that abnormal glomerular filtration and increased vasopermeability cannot explain the findings. Babesial infection in the rat provides a useful model for the study of acquired C3 metabolic defects that have been observed in humans with immune complex diseases.


Release of tumor-associated antigens by murine melanoma cells. Synthesis and release of radiolabeled macromolecules and tumor-associated antigens (MAA) by murine B16 melanoma was studied by pulse labeling cells in culture with 3H-leucine. Approximately 36% of newly synthesized macromolecules and 44% of newly synthesized MAA were released in 48 hr. MAA release was slightly, but consistently, more rapid than the average release of other macromolecules. Release of MAA did not result solely from cell death since it was greater than that of 51Cr-labeled molecules and cell viability was over 98%. The rate of release of newly synthesized MAA was not significantly influenced by cell replication. However, synthesis of MAA was much greater during the logarithmic than the stationary phase of cell growth, suggesting a concomitant increase in the amount of MAA available for release. These findings indicate that antigens and other macromolecules can be rapidly released by viable tumor cells.


Pathogenetic mechanisms in immune polioencephalomyelitis: quantitative evaluation of protective and pathogenic effects of lymphoid cells. Terminal dilution, adoptive cell transfer techniques were developed to quantify the protective effect of lymphoid cells in the pathogenesis of immune polioencephalomyelitis (IPE). The pathogenic effects of lymphoid cell populations were quantified by deleting the step of antigenic challenge. Regression curves were computer analyzed and PD50 values were compared. Immune spleen cells (ISC) from 4- to 6-week-old donors were more protective (PD50 = 4.9 +/- 1.3) than ISC from 12-month-old animals (PD50 greater than 7.0). The slopes of the regression curves also differed markedly (young mice, -0.24; old mice, -0.09). ISC were less protective in 12-month-indicator mice than in 5-month-old recipients (PD50 values of 5.2 +/- 0.8 and 3.7 +/- 0.8, respectively). When adoptive cell transfer tests were used to quantify the pathogenetic effects of donor cells it was found that ISC were pathogenetic at doses of 10(5) or less, but protective at higher doses. IPEC were pathogenetic at all test doses. When ISC were x-irradiated or sonicated the were only pathogenetic. Normal spleen or peritoneal exudate cells were neither protective nor pathogenetic. A model was developed in which mice were either thymectomized at birth (Tx), or Tx at birth and x-irradiated (500 R) 8 weeks later (Tx-XR). Sham Tx or Tx-XR mice served as controls. All of the mice were challenged with antigen (10(4) x-irradiated Ib cells). Only a portion (8/24) of the Tx mice developed IPE, indicating that resistance was T cell dependent but also involved a significant T cell independent component. The data also indicated that T cells were not pathogenetic effector cells in this model. Tx mice were not reconstituted by ISC (7/18 developed IPE), Tx-XR mice were partially reconstituted (3/12 developed IPE), but sham Tx-XR were fully restored (0/20 had IPE). Normal spleen cells did not reconstitute any of the mice.


Suppressor cells in the spleens of tumor-bearing mice: enrichment by centrifugation on hypaque-ficoll and characterization of the suppressor population. Spleen cells from mice bearing methylcholanthrene-induced sarcomas or a mammary adenocarcinoma suppressed the mitogen responses of normal spleen and lymph node cells. Lymph node cells from tumor bearers had no suppressive effects. Centrifugation of spleen cells layered on Hypaque-Ficoll (specific gravity of 1.08) produced a dense fraction which pelleted and a light fraction which was retained at the Hypaque-Ficoll-medium interface. Suppressive activity was not found in either fraction of normal spleen cells. In tumor-bearer spleen cells suppressor activity was greatly enriched in the light fraction. Treatment of the suppressor fraction with anti-theta or anti-Ig serum and complement did not remove suppressor activity. However, the suppressor cells were removed by passage through nylon wool or by carbonyl iron treatment. Also, the population which adhered to plastic Petri dishes contained the suppressor cell activity.


Antibody against the antigen receptor of a plasmacytoma prolongs survival of mice bearing the tumor. A transplantable plasmacytoma, TEPC-15 in BALB/c mice, induces a transient antibody response directed against the combining region of the myeloma protein produced by the tumor. Mice immunized with mitomycin-treated tumor cells produce similar antibody but do not develop tumor; mice so immunized survive longer than untreated mice when inoculated with viable TEPC-15 cells. This protection afforded by immunization can be transferred by serum alone; the protective effect of passively given serum is eliminated by absorbing out antibody directed against the myeloma protein.


Isolation and partial characterization of SAA-an amyloid-related protein from human serum. A 12,000 dalton serum amyloid A protein (SAA) has been isolated by chromatography on Sephadex in 10% formic acid. It is similar immunologically to the previously characterized 8500 dalton tissue amyloid A (AA) protein. The results of amino acid analyses, peptide maps, and the identity of the first 11 residues of the SAA and AA proteins support the idea that AA represents the amino terminal fragment of SAA and is derived from it by proteolysis.


The C5b-9 complex: subunit composition of the classical and alternative pathway-generated complex. The membrane attack complex of complement (C5b-9) is identical in composition regardless of which pathway of activation was instrumental in its formation. Band V protein was consistently a subunit of the soluble complex. Since band V protein is not required for complement-dependent cytolysis, it probably represents a membrane site equivalent in serum of the nascent C5b-9 complex.


Redistribution of the Fc receptor on human blood monocytes and peritoneal macrophages induced by immunoglobulin G-sensitized erythrocytes. The Fc receptor is a plasma membrane component exhibiting an affinity for the C gamma 3 domain of certain subclasses of immunoglobulin G. Using anti-Rho (D)-sensitized red cells (EA) in a slide rosette technique, we have demonstrated a translational mobility and polar redistribution of this receptor on the human blood monocyte and peritoneal macrophage. These cells, isolated from venous blood and malignant ascites and identified histochemically, showed a time- and temperature-dependent receptor capping defined by binding six or more EA confined to the cell half-perimeter. Caps formed in serum were mainly extreme caps in which bound EA appeared as a morula contiguous with the adherent cell. At 21 degrees C and 37 degrees C in serum 80% live rosettes formed caps; virtually none formed at 4 degrees C and about 25% were seen in PBS at 21 degrees C. Similarly, 10% extreme caps in PBS and 60 and 70% in serum were seen at room temperature and 37 degrees C, respectively, suggesting a serum factor(s) was important in promoting live rosette capping. Capping was reversibly inhibited by sodium azide although inhibition was incomplete even at 0.1 M, a concentration 10-fold higher than that giving complete inhibition of lymphocyte antigen-receptor capping. Azide also induced reversion of capped rosettes to diffuse, noncapped rosettes, and to levels comparable to those seen in inhibition studies. Re-exposure to EA after rosette capping failed to increase either the proportion of cells forming rosettes or the fullness of such rosettes indicating a critical number of receptors had been capped. Live rosettes induced a spherocytosis in bound EA irrespective of capping status; such changes appeared early in PBS where capping was minimal. Dead cells bore EA as normal biconcave discs. Some rosetting EA were ultimately hemolyzed upon prolonged incubation, and erythrophagocytosis was seen in about 15% of capped rosettes at 37 degrees C.


Motheater: an immunodeficient mouse with markedly less ability to survive that the nude mouse in a germfree environment. The motheaten mouse is an immunodeficient mutant which has been found to have elevated immunoglobulin levels, to exhibit autoimmunity, and early death. Since survival is not improved by a germfree environment, it appears that some mechanism other than infection results in its early demise.


Radiolabeling of immunoglobulin without loss of antibody activity--use of 14C-phenylisothiocyanate with human cytotoxic antibody. Radioiodination of immunoglobulins, particularly human cytotoxic IgG, induced a marked loss of antibody activity. Phenylisothiocyanate (PITC) readily reacts with alpha-amino groups and the epsilon-amino groups of lysines to form phenylthiocarbamyl derivatives. Since PITC is commercially available with 14C, 3H, and 35S substitutions, it provided an alternative means for labeling immunoglobulin which preserved antibody activity. There were approximately 80 PITC binding sites per IgG molecule, and 14C-PITC was bound with an efficiency in excess of 80%. With as many as 40 PITC molecules bound per IgG molecule, full cytotoxic activity was retained by both anti-HLA isoantisera and human anti-melanoma autoantisera. Even with 70 to 80 molecules of PITC bound per IgG molecule, over 80% of the antibody activity remained. Radiolabeling of antibody with 3H, 14C, or 35S-substituted PITC will greatly facilitate studies of antibody binding, particularly to cell surface antigens.


Receptors for immune complexes on cells within a non-lymphoid murine tumor. In this paper we present evidence that antibodies unrelated to the tumor cell can comprise part of the in vivo Ig surface coat of cells derived from the non-lymphoid murine tumor, TA3/St. This was shown by incubating TA3 cells originating from other OA or BSA preimmunized mice with radioiodinated 125I-OA and 131I BSA. Radioiodine-labeled 125I-OA specifically fixed to cells derived from TA3/St tumors originating from OA-preimmunized mice. On the other hand 131I-BSA was specifically fixed by cell populations from BSA-preimmunized mice. Incubation of these cells in vitro at 37 degrees C abolished the specific binding and antibody could subsequently be detected in the tissue culture medium. Radioiodine labeled purified soluble antibody-antigen complexes could also be bound to cells derived from freshly harvested TA3/St tumors but not to their in vitro propagated counterparts. Removal of phagocytic or adherent cells from these cell populations decreased the binding of the complexed antibody on freshly harvested TA3/St populations, but did not eliminate it. Inhibition of complexed antibody binding was obtained when TA3/St cells (an H-2a tumor) were pre-incubated with anti-H2a antiserum. Propagation of the tumor in an F1 hybrid (A X C57BL) in which host cells could be distinguished from tumor cells by using an anti H-2b antiserum showed that binding of the immune complex was mostly limited to host cells infiltrating into the tumor population.


The formation of septate-like junctional complexes between lymphoid cells in vitro. During the initial 48 hr of incubation in immunized Mishell-Dutton spleen cell cultures, most lymphocytes exist as single cells or an occasional pair of cells (doublets). Ultrastructural examination of the area between those cells forming doublets revealed a septate-like junctional complex occurring over large portions of the plasmalemma. This junctional zone persisted in areas where extensive cytoplasmic interdigitation among cell processes was evident. Approximately 1% of the lymphocytes isolated from the top fraction of a discontinuous fetal calf serum gradient were involved in formation of doublets. The majority of these doublets showed evidence of junctional interaction. Moreover, this septate-like zone of adhesion was seen using several different fixatives and stains and remained intact after osmotic cell disruption. The junction was not demonstrable by using freeze-fracture techniques and therefore must be limited to interactions within the surface of the cells.


Purification of homogeneous murine immunoglobulins with anti-fructofuranan specificity. Homogenous IgAK immunoglobulins ABPC-47 new, EPC-109, UPC-61 (anti-2,1 fructofuranan specificity) and homogeneous IgAK ABPC-48 and IgGK UPC-10 (anti-2,6 fructofuranan specificity) were purified by affinity chromatography by using inulin- and grass levan-Sepharose columns, respectively.


Mononuclear cells migration inhibition and delayed hypersensitivity in man. The migration inhibition by soluble PPD of peripheral blood mononuclear cells showed a much better reflection of delayed hypersentitivity to tuberculin in man than the migration inhibition of buffy coat cells. The migration index (MI) with mononuclear cells in a group of tuberculin positive subjects was 0.51 +/- 0.14 (mean +/- S.D.) (n = 20) and that of a group of tuberculin negative subjects was 0.89 +/- 0.11 (n = 14) (P less than 0.001). Using buffy coat cells the MI was 0.65 +/- 0.26 (n = 19) for a group of tuberculin positive subjects and 0.76 +/- 0.24 (n = 10) for the group of tuberculin negative subjects. They were not significantly different (P greater than 0.05). The mononuclear cells migration inhibition was also quite reproducible from day to day.


Development of an enzyme immunoassay for human placental lactogen using labelled antibodies. An immunoassay for the determination of human placental lactogen (HPL) has been developed using specific antibodies labelled with horseradish peroxidase. A method is presented in which the enzyme : antibody conjugate is purified with an antigenic solid phase to ensure the production of conjugates with intact immunological activity. Serum components interfere with the assay but this problem can only be avoided at the expense of a precise and practical assay for the determination of HPL.


The antigen-binding capacity of serum IgG. An immunosorbent method applied to subtilisin as antigen. A method for estimating the antigen-binding capacity of serum IgG is presented. A Sepharose anti-human IgG immunosorbent is incubated with test serum and washed. The antigen-binding capacity of the bound specific IgG is determined by incubation with purified, inactivated 125I-subtilisin. The method was applied tp the sera of workers in the detergent industry exposed to enzymatic preparations from Bacillus subtilis. The sera of 33 out of 69 workers bound a detectable amount of subtilisin, the detection limit ranging from 8--34 ng/ml in different assays. The amount of antigen bound ranged from 42--9380 ng/ml. IgG to subtilisin was not detected in the sera of 61 subjects with no known exposure to subtilisin and who were not factory workers.


Effects of sulfate concentration, electroendosmotic flow, and electrical resistance of agars and agaroses on counterimmunoelectrophoresis with adenovirus antigens and antisera. The sulfate concentration, electroendosmosis (EE), and electrical resistance of 23 commonly used agars and agaroses were measured by standard techniques and related to counterimmunoeletrophoresis (CE) with adenovirus antigens and antisera. Statistical analyses of the test procedures themselves showed that the tests had sufficient reproducibility to warrant further analyses on the interaction or combined effects of these parameters on adenovirus CE tests. Both sulfate concentration and EE were related to CE by cubic polynomial equations; sulfate was directly related to EE by a linear equation. Resistance was somewhat linearly related to CE, but the differences among agars were too small to show a clear correlation. The data show that the fixed charges of the sulfate anion are major, although probably not the only, contributors to EE. The combines EE calculations from spectrophotometrically-scanned and hand-measured slides were used to predict the usefulness of new lots of agaroses for diagnostic adenovirus CE tests, according to the derived equation, CE score = 4.9 + 112.1 EE --397.1 EE2 + 329.1 EE3. The best stabilizing media for adenovirus CE tests had sulfate concentrations of 0.37% +/- 0.16% and EE cathodic mobility ratios of 0.17 +/- 0.03.


Selective action of hydrocortisone on postmitotic epidermal cells in vivo. Hydrocortisone administered systemically for 3 weeks has no effect on any phase of epidermal cell proliferation as measured by autoradiographic methods. However, the speed of cell differentiation (maturation) is increased, resulting in a thinning of the living epidermis due to the shorter epidermal cell life. Comparison of the epidermis from two body sites (ear and sole of foot) in mice receiving 2.4 mug per gm body weight per day of hydrocortisone in drinking water for 3 weeks revealed no change in the labeling with [3H]thymidine, the mitotic indices, or the lengths of the cell cycle phases. Quantitation of the epidermal cell compartments showed that thinning of the epidermis with hydrocortisone was due to the loss of an identical number of differentiating epidermal cells per unit surface from both body sites. In both sites there was the same increased rate of maturation of postmitotic cells while the proliferative cell-pool remained unresponsive to the hormone. The alteration of the speed of cell maturation is the principal action of hydrocortisone in epidermis. The results indicate that the epidermal cellular concentration of, and the susceptibility to, the hormone were identical in ear and sole of foot despite the differing speeds of turnover of the two tissues.


Cytophotometric studies of epidermal proliferation in psoriatic and normal skin. Cytophotometric measurements of single-cell DNA content were used to study human epidermal cell proliferation in vivo. It was found that Feulgen-DNA profiles can be used to assess proliferative activity in involved, uninvolved, and nonpsoriatic skin. Profiles of involved psoriatic skin were bimodal as is characteristic of actively proliferating populations. This was due to the presence of cells with twice (2C--presynthetic) or four times (4C--post-synthetic) the amount of DNA of the gametes, separated by the intermediate values of cells undergoing scheduled DNA synthesis. Profiles of uninvolved psoriatic as well as nonpsoriatic epidermis were unimodal with the majority of cells containing a 2C amount of DNA incating relatively low levels of proliferative activity. The observed variations in proliferative activity of these samples are discussed in terms of two alternative models. Since radioisotopes are not required, this technique presents a useful approach to studying human epidermal proliferation in vivo.


Ultraviolet light carcinogenesis in hairless mice: cell kinetics during induction and progression of squamous cell carcinoma as estimated by the double-labeling method. The double-labeling method for studying cell cycle kinetics was applied to uninvolved epidermis of hairless mice (irradiated and unirradiated), and at various stages of ultraviolet light (UVL)-induced squamous cell carcinoma. The growth fraction (GF) was determined by continuous labeling with tritiated thymidine (3HTdR). The data indicate that the double-labeling technique is an acceptable method for studying cell kinetics in UVL-induced squamous cell carcinoma. The GF in normal epidermis (unirradiated and irradiated), hyperplastic epidermis, and lesions of squamous cell carcinoma is 1. The duration of cell generation (Tc) and DNA synthesis (Ts) periods decreased progressively with induction and progression of squamous cell carcinoma. For normal epidermis (irradiated mice) the Tc and Ts were 91 and 6.3 hr, respectively. In the hyperplastic epidermis, the Tc and Ts were reduced to 44 and 4.5 hr, respectively, while even lower values were obtained for the tumor (Tc = 16 hr, Ts = 3.1 hr). The labeling index (LI) in normal epidermis was 7% and increased progressively in hyperplastic epidermis (10%) and squamous cell carcinoma (19%). Epidermal cell differentiation in hyperplastic and tumor tissues appears to have been delayed (GF = 1), since cells above the basal layer, which in normal epidermis keratinize, retain the ability to proliferate, as evidenced by extensive incorporation of [3H]TdR in these cells. The results suggest that tumor production is associated with a progressive shortening of the cell cycle and delayed keratinization of epidermal cells.


A liquid film model of tetrakaidecahedral packing to account for the establishment of epidermal cell columns. The possiblity that the organization of cells into columns in the mammalian epidermis may be a result of the close packing of these cells has been investigated in a model system involving the association of randomly produced soap bubbles into a stable froth. Upon floating to the surface of a liquid, soap bubbles have been found to spontaneously assemble into precise columns of interdigitating bubbles. The tetrakaidecahedral shape and the spatial configuration of these bubbles closely resemble those of stacked epidermal cells, although the columns of a froth were oriented at a 60degrees angle to their substratum rather than at right angles as occurs in the epidermal cell columns. These observations lend support to the theory that the organization of the cells in the epidermis into columns is due to the assumption of the keratocytes of a minimum surface-close packing array. Such an organizing mechanism would be independent of both positional control of the underlying mitoses and active guidance of the cells as they become superficially displaced within the epidermis. The observation that a high rate of cell turnover is incompatible with the epidermal column structure may be related to the finding that rapidly produced soap bubbles do not, at least initially, assemble into a columnar array.


Natural synchrony of newborn mouse epidermal cells in vitro. Epidermal cells were separated from newborn mouse skin and grown on glass chamber slides or in plastic Petri dishes. Cell proliferation was examined at short intervals during the first 3 to 4 days in culture. DNA synthesis was estimated by the incorporation of [3H]thymidine into epidermal DNA, or by the labeling index. The mitotic rate was estimated by blocking mitotic cells with vinblastine. Reasonable agreement was found with the different methods. The cells grew synchronously with peaks of DNA synthesis at 21 to 23 hr, 35 to 40 hr, and 60 hr; peak mitotic rates were seen at 44 to 48 hr and 72 to 76 hr. The cells appeared to grow exponentially during the first 3 to 4 days in culture, with one main cohort of cells dividing during the successive proliferative peaks.


Freeze-fracture of Langerhans granules. A comparative study. A comparative study of freeze-fracture replicas and thin sections of Langerhans granules has proved that the effects of conventional tissue processing on the inner structure of the granule are negligible, and the study has confirmed the validity of the previously proposed tridimensional models. The paracrystalline quality of the substance contained between the limiting membranes of the disc-shaped area of the granules seems to suggest it protein nature. In replicas, no particle organizations attributable to intercellular junctions were observed on the plasma membranes of Langerhans cells.


Organ culture of adult mouse esophageal mucosa in a defined medium. A method is described for culturing adult mouse esophageal mucosa in a chemically defined medium. The preparation consists of mucosa and superficial submucosa. By light microscopy, autoradiography, and tritiated-thymidine [3H]TdR) uptake the tissue appears viable for at least 3 days. Although a drop in the rate of [3H]TdR uptake is observed in the initial hours of culture, recovery occurs by 24 hr and uptake remains constant for at least an additional 48 hr. Twenty-four-hour exposure to [3H]TdR and autoradiography reveals that 93 +/- 3% of the basal cells take up label; appreciable labeling is not found in other cells of the preparation. Pulse labeling indicated a transit time from basal layer to keratin layer of about 72 hr. This preparation should be useful for short-term in vitro studies of keratinized, stratified, squamous epithelium free of appendages and for studies of the growth properties of esophageal mucosa under simulated pathologic conditions.


Importance of pyrimidine nucleotide salvage pathways for DNA synthesis in skin. Split-thickness rabbit skins were minced and incubated in vitro with radioactive precursors selected to measure do novo and salvage pathways for pyrimidine nucleotide synthesis. In this system, both the salvage precursors [3H]thymidine and [14C]cytidine were incorporated actively into skin DNA and only [14C]cytidine into skin RNA. In contrast, the de novo precursor [14C]orotic acid labeled skin RNA extensively but did not significantly label skin DNA. An unusually high ratio of specific activities of UMP:CMP in RNA was observed when [14C]orotic acid was used as a nucleotide precursor. Methotrexate effectively blocked thymidylate synthesis do novo but did not inhibit DNA synthesis as determined by [3H]thymidine incorporation into DNA thymidylate residues or by [14C]deoxycytidine incorporation into DNA deoxycytidylate residues. Azauridine inhibited labeling of RNA by [14C]orotic acid to a greater extent than labeling of DNA by [3H]thymidine. These results suggest that in rabbit skin, cytidine nucleotides are utilized more effectively than uridine nucleotides for deoxypyrimidine biosynthesis, that DNA synthesis is primarily dependent upon salvage mechanisms to supply deoxypyrimidine nucleotides, and that inhibition of thymidylate synthesis by methotrexate does not inhibit DNA synthesis in this system.


Testing the effects of fluids on rat skin using disposable polypropylene chambers. Inexpensive, disposable polypropylene chambers for testing fluids on rat skin were made by removing the wings from polypropylene wing nuts and by cutting into the top of each nut a seat for an O-ring. Polypropylene screws with black heads were fabricated as seals for the chambers. After attachment to the backs of rats, these chambers provided a satisfactory  means of keeping fluid in contact with the skin for 3 days.


Lead content of tissues of baby rats born of, and nourished by lead-poisoned mothers. Three groups of Wistar rats (5 males and 5 females in each group) were given zero, 100 mg., and 200 mg. Pb(NO3) 2 per kilogram of body weight per day for 45 days and were mated. The lead feeding was continued throughout the duration of experiments. The blood and urinary lead, blood aminolevulinic acid dehydratase activity, and free erythrocyte protoporphyrin and urinary aminolevulinic acid were measured on these rats at the beginning of the study, at the time of mating, and prior to delivery. The tissues, incuding kidney, liver, heart, and brain of the newborn progeny were analyzed for lead content. The blood lead and aminolevulinic acid dehydratase were also measured (pooled blood). These tissues contained very high amounts of lead when the newborns were killed within half an hour after their birth, a very significant reduction being found after one day with further reduction after 7 days. Kidney and heart contained the highest amount of lead, the levels being dose-related. Three weeks after birth the lead content of tissues rose significantly but blood concentrations remained almost identical to the levels obtained 1 day after birth. There was no evidence of lead accumulation in the brain of the babies...


Molecular size of rabbit C-reactive protein synthesized in vitro. This study was performed to determine whether immunologically detectable C-reactive protein synthesized by rabbit liver mince in vitro is a micro- or macromolecular precursor of CRP, or if it is native CRP. Employing gel filtration and density gradient techniques, the molecular size of CRP newly synthesized by minced rabbit liver from inflamed rabbits was found identical with that of native CRP. Since no evidence for hepatic synthesis of a precursor can be shown, increased CRP formation after inflammation must result from increased de novo synthesis.


The relationship of serum ionized and total calcium in primary hyperparathyroidism. Ionized and total calcium concentrations were measured in the same serum samples of 48 patients with primary hyperparathyroidism in order to compare diagnostic accuracy and sensitivity and to determine the intra-individual variation of these parameters in this disorder. In 149 samples, 0.7 per cent of serum ionized calcium and 7.4 per cent of total calcium determinations were within the normal range (mean +/- 2 S.D.). However, all patients exhibited abnormally elevated values upon repeated testing. Ionized and total calciums were significantly correlated both preoperatively (r=0.85 and p less than 0.001) and postoperatively (r greater than 0.9). Heterogeneous and abnormally large variations of both measurements were noted in these patients. It is concluded that the superior precision of the method for ionized calcium determination renders it more accurate than total calcium for the diagnosis of primary hyperparathyroidism. The disturbance of calcium homeostasis is reflected in both calcium and its ionic fraction in serum.


Effect of acetazolamide on sodium reabsorption in the diluting segment in man. The purpose of this study is to compare the effects of extracellular volume expansion (ECVE) and acetazolamide (ACTZ) on the transport of sodium in the diluting segment in man. All studies were performed at the peak of sustained oral water diuresis (Uosm. less than 75 mOsm. per kilogram). Distal delivery was estimated by the rate of urine flow and distal sodium reabsorption was approximated by the generation of free water. Free water clearance for given rates of distal delivery expressed as V(range 10 to 35 ml. per minute) was significantly lower and the clearance of sodium higher in the ACTZ group as compared with subjects with ECVE. However when CH2O was plotted against distal delivery expressed as CC1 + CH2O, no difference was detected between ACTZ and ECVE studies. We interpret these findings as suggesting that ACTZ does not have a direct inhibitory influence on the NaCl pump at the diluting site. Rather, the lowering of CH2O/V following ACTZ is due to the flooding of the diluting site with the poorly reabsorbable bicarbonate ion. Thus, the proximal action of ACTZ, which initiates urinary bicarbonate loss results secondarily in an apparent interference with the diluting mechanism in man.


Correction of protein binding defect in uremic sera by charcoal treatment. Protein binding of numerous drugs, primarily organic acids, is decreased in sera from uremic patients. The defect in binding is (1) greater than can be accounted for by hypoalbuminemia alone; (2) unchanged by prolonged in vitro dialysis; (3) transferred in the protein but not the ultrafiltrate fraction of uremic serum; and (4) not reproduced by addition of low and middle molecular weight compounds known to accumulate in uremia. However, treatment with activated charcoal at pH3 was found to significantly increase drug protein binding in uremic sera. This effect was studied with six different drugs in sera from groups of 6 normal subjects and 8 patients on chronic hemodialysis. The percentage of sulfamethoxazole, dicloxacillin, diphenylhydantoin, salicylate, and digitoxin bound to protein in normal sera (65.9, 97.1, 93.1, 96.7, and 92.7, respectively) was unchanged by charcoal treatment. In contrast, charcoal treatment significantly (p less than 0.01) increased the percentage of drug bound to protein in uremic sera from 41.7 to 59.0 for sulfamethoxazole, 90.7 to 96.3 for dicloxacillin, 84.3 to 90.8 for diphenyhydantoin, 86.4 to 93.8 for salicylate, and 89.5 to 90.9 for digitoxin. Charcoal treatment significantly (p less than 0.05) reduced penicillin protein binding in normal sera and failed to correct the binding defect for penicillin in uremic sera. The effect of charcoal can be explained by removal of an inhibitor which accumulates in uremia and (1) occupies the binding site of certain drugs, (2) changes the configuration of the albumin molecule, or (3) both. Free fatty acid (FFA) concentrations in uremic patients were similar to those in normal subjects and were not the cause of the binding defect.


A new iron-binding protein isolated from intestinal mucosa. A new iron-binding protein contained in guinea pig intestinal mucosa has been purified to homogeneity. The protein has a molecular weight of 78,000 in a nondissociating system and 43,000 in SDS-gel electrophoresis. It binds approximately 2 moles of iron per mole with a formation constant of 10(19) at pH 7. It is distinguished from transferrin and lactoferrin by differences on DEAE-Sephadex chromatography and spectroscopy and by its failure to cross-react with antisera to these other iron-binding proteins.


Effect of mannitol on phosphate transport in intact and acutely thyroparathyroidectomized rats. Clearance experiments were performed in male Sprague-Dawley rats to determine the effect of mannitol on phosphate (Pi) transport. Solutions of 10 per cent mannitol or normal saline were infused at progressively increasing flow rates with or without parathyroid extract (PTE) infusion into the following animals: Group I: Intact Rats-Mannitol Infusion: A--Intact, hypocalcemic; B--Intact, normocalcemic, normomagnesemic; Group II: TPTX Rats: A--Mannitol infusion; B--Mannitol + PTE infusion; C--Hydropenia + PTE infusion; and D--Saline + PTE infusion. In contrast to previous reports, mannitol increased Pi excretion in intact rats. When Ca + Mg were maintained constant in intact rats or after TPTX, mannitol failed to increase Pi excretion. In TPTX rats receiving mannitol + PTE, increased Pi excretion was again noted. Comparison of Pi excretion during PTE infusion during hydropenia and volume expansion with mannitol or saline in TPTX rats revealed significantly higher Pi excretion with volume expansion. Pi excretion paralleled Na excretion in intact mannitol-loaded and PTE-infused TPTX animals undergoing a mannitol or saline diuresis. Pi and Na excretions, however, were dissociated in mannitol-loaded TPTX rats, intact animals receiving simultaneous Ca and Mg infusion, and TPTX hydropenic animals receiving PTE. These studies indicate that (1) mannitol increases Pi excretion in intact rat, (2) the phosphaturia is PTH-mediated, (3) Pi and Na excretions can be dissociated, and (4) volume expansion with either mannitol or saline enhances the effect of PTH on Pi transport in the renal tubule.


Chemotherapy of experimental streptococcal endocarditis. V. Effect of duration of infection and retained intracardiac catheter on response to treatment. The dynamics of infection and response to penicillin G (P) or combined P plus streptomycin (S) therapy were studied in a rabbit model of penicillin-sensitive Streptococcus sanguis endocarditis. Increased delay between infection and initiation of treatment required longer duration of therapy to effect a cure. Irrespective of the interval between infection and onset of therapy, P combined with S regularly eliminated streptococci from endocardial vegetations more rapidly than P alone, both in the presence and absence of an intracardiac catheter--although the difference in response was more marked with the catheter present. The combination of P and S consistently showed superior in vivo bactericidal activity compared to P alone in established S. sanguis endocarditis.


Inhibition of platelet aggregation by 5-nitro-2-furaldehyde diacetate with observations on structure-activity relationships. Nitrofurantoin [1-(5'-nitro-2'-furfurylideneamino)-2, 4-diketoimidazole] is a potent inhibitor of primary ADP-induced platelet aggregation. The nitro group on the furan ring and the specific arrangement of the two keto groups on the imidazole ring are the molecular characteristics of nitrofurantoin critical to its inhibitory effect. The present studies report that 5-nitro-2-furaldehyde diacetate is also a potent inhibitor of primary ADP-induced platelet aggregation. 5-Nitro-2-furaldehyde diproprionate, 5-nitro-2-furaldehyde dibutyrate, and 5-nitro-2-furfuryl acetate are essentially inactive. These results indicate that a diacetate moiety can replace the diketoimidazole moiety of nitrofurantoin and form a compound that fully retains the inhibitory effect upon primary ADP-induced platelet aggregation.


Drift in the oxygen content measured with the Lex-O2-Con: long-term assessment. After more than one year of intensive use (30 to 50 measurements a day) a drift was observed in the data given by two Lex-O2-Con analyzers. This was confirmed by iterative comparisons with the Van Slyke method. After recharge of the cells, the instruments seem to function again correctly.


A radioimmunoassay to determine the subtypes D and Y of the hepatitis-B surface antigen. A radioimmunologic test system for determining the subtypes D and Y of the hepatitis-B surface antigen (HBs-Ag, Australia antigen, HAA) is described. This technique uses the immunoadsorption principle. The antigen to be subtyped is incubated with an antiserum which contains (125)I-labeled antibodies to the antigenic determinants a, d, and y. The specificity of the antibody which is not consumed in forming the antigen-antibody complex is determined by absorption on agarose to which HBs-Ag of subtypes D and Y is coupled. This specificity permits conclusions as to the subtype of the hepatitis-B surface antigen which is being investigated.


Effect of simultaneous thiamin and riboflavin deficiencies on the determination of transketolase and glutathione reductase. Thiamin and/or riboflavin status have been assessed in rats simultaneously deprived of both vitamins and subsequently repleated with either one or the other in succession. Undernutrition in control rats was found to depress the activity of the erythrocyte enzymes (transketolase and glutathione reductase, respectively) used to assess status. On the other hand, the activity coefficients obtained by in vitro stimulation of the enzymes with their respective missing cofactors were not suppressed. Therefore the activity coefficients were superior indices of thiamin and riboflavin nutriture compared to the enzyme activities. When activity coefficients were used as the indices of vitamin status there did not appear to be interference from the simultaneous deficiency of the alternate vitamin. Apparent interference was encountered when the enzyme activity itself was used as the criteria.


The effect of uremia on penicillin flux between blood and cerebrospinal fluid. The effect of uremia on the choroid plexus system responsible for transport of penicillin from cerebrospinal fluid (CSF) to blood was studied in vitro and in vivo. Uremia was induced in rabbits by injecting toxic doses of cephaloridine or by obstructing urine flow. Three days after the induction of uremia, isolated choroid plexuses from normal rabbits were unable to concentrate penicillin 14C normally when incubated in either CSF or ultrafiltrates of plasma from uremic rabbits. In vivo, a decrease in the disappearance of penicillin 14C from CSF was observed in uremic rabbits. However, the choroid plexus transport system for penicillin was only partially and reversibly depressed in uremia. The increased CSF levels of penicillin uremia are due to: decreased excretion of penicillin by the kidney, depression of active efflux of penicillin from the CSF, and decreased plasma binding of penicillin.


A new approach to quantitation of the various sources of bilrubin in man. Studies of the "early labeling" of fecal bile pigment have been widely interpreted as indicating that 10 to 20 per cent of total bile pigment production in normal man is derived from the processes which give rise to the "early labeled" pigment peak (ELP). However, this conclusion is based on a very small number of studies with inherent experimental and analytic limitations. In the present study, a new experimental approach was employed. Plasma bilirubin turnover (BRT), total red blood cell volume (TRCV), and red cell survival were measured in 26 normal volunteers and 35 patients with various hepatic and/or hematologic disorders. The quantity of bilirubin derived from red blood cell degradation (BRRBC) was calculated from TRCV and the mean red cell life-span, and the difference between BRT and BRRBC, designated "excess bilirubin synthesis (EBS)," was considered to represent the quantity of bilirubin derived from the processes responsible for ELP. In normal volunteer subjects, EBS averaged 1.0 mg. per kilogram per day, and accounted for 25 per cent of daily bilirubin turnover. Hence, results derived by this approach exceeded the range estimated from analysis of "early labeled peak" data. Further analysis of the data in terms of a simple model of bilirubin sources was compatible with the hypothesis that EBS consists of an essentially constant component, averaging 0.8 mg. per kilogram per day, presumably derived principally from the liver, and a second component which varies with the erthropoietic rate. Hence, in normal man the liver may contribute the major fraction of EBS.


Changes in the ratio of plasma to intersitiial fluid volume during long-term antihypertensive therapy. Based on determinations of plasma (T-1824)(PV) and extracellular fluid volumes (82Br- distribution space) (ECFV), the PV/IF ratio (IF=ECFV.PV) was calculated three times with a 1-week interval between determinations in untreated patients with essential hypertension. During treatment with hydrochlorothiazide (13 males and 8 females), hydrochlorothiazide-triamterne (5 males and 5 females), hydrochlorothiazide-alpha-methyldopa (7 males and 4 females), and propranolol (6 males and 4 females), PV/IF was determined after 2 to 6 months of treatment. PV/IF was significantly reduced during hydrochlorothiazide treatment (delta PV/IF:--0.011, S.D.: 0.011, p is less than 0.001) and propranolol treatment (delta PV/IF:--0.017, S.D.: 0.022, p is less than 0.05), remained unchanged during hydrochlorothiazide-triamterene treatment and increased significantly during hydrochlorothiazide-alpha-methyldopa treatment (deltaPV/IF: .013, S.D.: 0.015, p is less than 0.05). There was a negative correlation between PV/IF and mean blood pressure during hydrochlorothiazide treatment (r:--0.49, n: 21, p is less than 0.05 (and hydrochlorothiazide-triamterene treatment (r:--0.79, n: 10, p is less than 0.01), whereas no correlation was found during treatment with hydrochlorothiazide-alpha-methyldopa or propranolol. Correlation between mean blood pressure and the individual volumes was not observed nor was there any correlation between changes in the parameters investigated. It is concluded that PV/IF is more accurately correlated to mean blood pressure than are the individual volumes and that, during long-term antihypertensive treatment, PV/IF will reflect changes in the tone of the capacitance vessels and the venous resistance vessels, resulting in a changed equilibrium between the intravascular and the interstitial volumes. It is suggested that the reduction in PV/IF during thiazide and propranolol therapy is related to a relative increase in venoconstrictor sympathetic activity whereas the increase in PV/IF by alpha-methyldopa is caused by decreased sympathetic tone and venodilation.


Vitamin B6 status of young men, women, and women using oral contraceptives. Vitamin B6 intakes of 33 males, 73 females, and 46 females using oral contraceptives, all 18- to 25-year-old students, were compared to coenzyme stimulation of erythrocyte alanine aminotransferase activities of blood samples obtained from these subjects in order to ascertain the vitamin B6 status and perhaps the requirements of this population group. Trained interviewers obtained 24-hour recalls; subjects supplied 2-day food records. Dietary record analyses revealed that students reported consuming slightly less kilocalories, more protein, and considerably less vitamin B6 than the 1974 recommended dietary allowances. Erthrocyte alanine aminotransferase activities, with and without additional coenzyme, of blood samples from the 3 groups were significantly different from each other; per cent coenzyme stimulation of enzymatic activity of females and females on oral contraceptives were significantly different from each other. Enzyme data were not significantly affected by race, age, income, food budget, preparation of own food, frequency of eating out, or vitamin supplementation. Several subjects in all 3 groups had high per cent stimulation values which is considered to be indicative of subclinical vitamin B6 deficiency. Subjects with values near 0 per cent stimulation had varying vitamin B6 intakes thus perhaps indicating large individual variations in vitamin B6 requirements.


Determinants of deranged sodium and water homeostasis in decompensated cirrhosis. Previous studies from this laboratory have demonstrated that the redistribution of blood volume and concomitant central hypervolemia induced by water immersion to the neck (NI) results in a significant natriuresis, kaliuresis, and diuresis. The NI model was utilized to assess the role of "effective volume" and hyperaldosteronism in the impairment of sodium and water handling in cirrhosis. Eleven cirrhotic patients were studied twice while in balance on a 10 mEq. Na, 100 mEq. K diet: control and NI. The conditions of seated posture and time of day were identical. UNaV was constant throughout C, ranging from 1 to 2 muEq per minute. During NI, UNaV increased progressively from 1 +/- 1 (S.E.M.) during the prestudy hour to 89 +/- 32 muEq per minute during hour 5 of NI (p less than 0.02), greatly exceeding the comparable value found in normal subjects on an identical diet. (See article).


Combined pre-immunization and granulocyte transfusion therapy for treatment of pseudomonas septicemia in neutropenic dogs. An experimental model was designed to evaluate a combined protocol of active immunization and granulocyte transfusions for treatment of Pseudomonas aeruginosa sepsis in the neutropenic host. One member of a pair of dogs was immunized with P. aeruginosa vaccine. Both dogs were then rendered transiently neutropenic with a single intravenous dose of cyclophosphamide (40 mg. per kilogram) and challenged with an intravenous inoculum of P. aeruginosa. Twenty-four and 48 hours after pseudomonas challenge each animal received granulocyte transfusions. Effectiveness of therapy was evaluated by observation of survival time, febrile response, and quantitative blood cultures. Results showed a significant increase in the survival period (P is less than 0.05), a lower febrile response (P is less than 0.025), negative blood cultures, and a greater recovery rate in the immune group. Immune dogs that died had negative blood cultures or less than or equal to 10 pseudomonas per milliliter of blood despite the presence of P. aeruginosa in tissues. In contrast, control dogs had septic deaths within 67 hours of pseudomonas challenge, marked febrile responses with 24 hours of infection, and positive blood cultures with 4,000 to 25,800 pseudomonas per milliliter of blood. These data show that combined therapy with immunization and granulocyte transfusions is effective in reducing the severity of P. aeruginosa infection and in preventing bacteremia during periods of leukopenia.


A three-month survival of a calf with an artificial heart. An artificial heart constructed from Biomer, a polyurethane, kept a calf alive for more than three months after its natural heart was removed. During this time all of the calf's vital organs apparently functioned well. We had been able to keep similar animals alive with Jarvik III hearts made from Silastic for one month. The principal problem encountered was the infection centering along the compressed air drive lines to the artificial heart. Another problem is thrombus formation in the artificial heart. Redesigning of the artificial ventricles to improve flow patterns and reduce material discontinuities should be attempted to eliminate localized thrombus formation within the ventricles. The rapid growth of the calf has led us to examine the possibility of alternative animal models for future long-term experiments. The most encouraging aspects of this experiment were the long survival time of the calf and the very low level of blood damage caused by the artificial heart.


Procoagulant and platelet aggregating properties of antilymphocyte sera. Ten anti-human lymphocyte sera in clinical use at six transplant centers and five anti-nonhuman lymphocyte sera were studied for their procoagulant and platelet-aggregating properties. This investigation was initiated following the observation of a clinical episode of renal artery thrombosis, associated with the use of one of the sera in a patient who had received a cadaveric renal transplant. The procoagulant from this serum shortened the clotting times of individual samples deficient, respectively, in factors XII, XI IX, X VIII, and VII. Esterolytic activity, demonstrated on the substrate benzoylarginine ethyl ester, was completely inhibited by phenylmethylsulfonyl fluoride, but coagulant activity was variably affected. The platelet-aggregating activity (PAA) has been identified as a complement-fixing gamma G antibody that was absorbed from the antilymphocyte serum (ALS) with human spleen lymphocytes. Five of the ten anti-human lymphocyte sera showed varying procoagulant activity (PCA), and six sera demonstrated PAA. Three sera contained both activities. The presence of both PCA and PAA in the same preparation may predispose patients to thrombotic events, particularly if administered intravenously. Sera should be screened for PCA and PAA prior to clinical use, and sera with these properties should be used with caution.


The measurement of rates of mineral loss with aging. The photon absorption technique was used to measure bone mineral content in the radius repeatedly with time to determine the feasibility of using this method in studies on the rate of loss with aging. Two groups of postmenopausal women were followed, a younger group for an average of 3.74 years and an older group for an average of 2.37 years. It was determined that estimates of rate over periods less than two years were too low in precision to be useful. There were significant differences in rates among individuals. The loss of mineral from the midshaft site was determined to be linear within the precision of the method and over an average interval of 3.74 years. From these data the size of groups and length of time needed for a study to determine the effect of a treatment upon inhibition of bone loss was predicted. When the inhibition is 50 per cent of the control rate, it would be possible to detect a significant (alpha=0.05) inhibition with a certainty of 95 per cent (1-beta = 0.95), if 56 subjects were followed in each group for a period averaging 2.70 years. A more potent inhibition or more frequent visits would reduce the number of subjects and duration of study needed. The separation of individuals into groups of significantly more rapid and slower rates of loss was demonstrated. From these data, it is possible to predict that one-third of a sample could be so characterized after 3.5 years. The photon absorption technique can be applied to the measurement of the rate of loss of mineral with aging with a precision that is sufficicient for certain types of clinical investigations.


A capillary test for the control of heparin therapy. The BaSon test (Baden, Sonnenfield) for the control of heparin therapy has been studied in capillary and venous blood of normal and heparinized children and the same reliability is found. The sensitivity to heparin and the reproducibility of the test have been observed in in invitro studies and the findings show that the sensitivity is excellent and the reproducibility is very good.


A semiautomated micromethod for dextran in the presence of glucose. A semiautomated method for microanalysis of dextran with anthrone reagent has been developed. The method utilizes the Technicon inulin manifold and other standard AutoAnalyzer equipment. Since the anthrone method is sensitive to plasma glucose, a glocuse oxidase method which eliminates glucose in samples to be analyzed for dextran has also been developed. When solutions containing 50 mg. per 100 ml. of dextran were compared to solutions containing 50 mg. per 100 ml. of dextran plus up to 200 mg. per 100 ml. glucose, no significant difference was observed. The coefficient of the anthrone-glucose oxidase method is 0.039, which is within the error of the AutoAnalyzer itself.


An enzymic method for the spectrophotometric determination of oxalic acid. A simple method is described for the measurement of urinary oxalate. Oxalate decarboxylase is coupled with NAD+ requiring formate dehydrogenase and the result recorded spectrophotometrically. Accurate determination can be carried out either on urine or a citrate extract or urine. Using the citrate extract procedure, the urinary samples can be stored for at least 3 months without any effect on the oxalate content.


Congenital aural and pre-auricular sinuses in the South African Bantu. The incidence of ear pits was studied in the South African Bantu population. A short account of the anatomy, occurrence, inheritance, developmental origin, complications and treatment of this anomaly was given. Mention has been made of the clinical significance and the possible association between ear pits and other systemic and ear abnormalities, including deafness. There seems to be a higher incidence of other abnormalities in children born with sinuses. The blanket term 'pre-auricular fistula' should possibly be discarded in favour of auricular sinuses or ear pits.


Audiological and haematological studies on the Todas of Nilgiris. A detailed audiological study combined with certain haematological investigations which were never previously done was undertaken on Todas, a small vanishing tribe of Nilgiris, South India. Ten per cent of the Todas studied exhibited otosclerosis which, though lower than that of a previous study, is still a very high percentage when compared to the general population. The incidence of chronic suppurative otitis media in Todas is markedly low (1-6 per cent) and 13-3 per cent of Todas had sensorineural hearing loss. Evaluation of hearing thresholds at various frequencies in the different age groups did not show much variation from that of the general population. The incidence of presbyacusis is not found to be low as reported earlier. Blood pressure recordings did not show any age-related increase. The majority of Todas studied belonged to the blood group 'B' (63 per cent) and 16-6 per cent belonged to the 'AB' group, which is distinctly different from the distribution pattern seen in the general Indian population. There was no significant variation in the serum cholesterol level in Todas.


Gunshot wounds to the head and neck.

Gunshot wounds to the head and neck. The types of injury which occur as a result of the civil disturbances in the north of Ireland are described. Four cases of gunshot wounds to the head and neck are described, each of particular clinical interest. The recent literature on the subject is reviewed and the consensus of opinion appears to be that the safest policy is to explore all cases of penetrating wounds of the neck


Traumatic pneumocephalus.

Traumatic pneumocephalus. A case is described of pneumocephalus involving both frontal lobes, following a frontoethmoidal fracture. It is noteworthy because it illustrates how extensive cerebral damage may be before it it detected. Intracerebral pneumocephalus occurs most commonly in fractures involving the frontal sinus or anterior ethmoidal/cribriform plate area. These fractures are not rare, and this potential complication should be in the minds of Ear, Nose and Throat surgeons who deal with trauma to the frontonasal region.


Oxygen tension and surface temperature of the endonasal tissues in the goat. Preliminary report. Measurement of the PO2 in the mucosal cells and at various depths of the lamina propria lining the nasal cavity in five male goats using Silver's special microelectrodes, revealed relative hypoxia in the anterodorsal part of the nasal septum and tips of naso- and maxilloturbinals. In these sites PO2 values varied between 55-65 at the surface and 30-35 in the superficial subepithlial layer while elsewhere in the nose, lip and tongue,  PO2 values of 85-90 were obtained. Temperature measurements with the thermistor in two goats yielded readings of 34-34-5 degrees C. in the hypoxic areas against  36-37 degrees C. in the posterior parts, the roof and floor of the nasal cavity. However, the oxygen tension diminished steadily in all areas as the microneedle electrode approached the chondro-osteal tissues but PO2 values of 0-10 in the perichondrium. The significance of these findings in the predilective localization of granulomata and various forms of vasculitis in the anterior endonasal tissues has been discussed.


Malignant lymphoma presenting as a parotid mass. Three cases of maglignant lymphoma presenting as a parotid mass have been presented. Head and neck surgeons should be aware of this condition both as a possible variant of lymphoma presenting in the head and neck region, and also as a part of the differential diagnosis of parotid gland enlargement. Early diagnosis is essential in these cases, as this has a beneficial effect on both the prognosis and management of these patients. The histopathology and staging of these tumors has been discussed with regards to the prognosis and management, and the difficulty of making the diagnosis and the chances to improve this are also discussed.


Myiasis in otorhinolaryngology with entomological aspects. Maggot infestation in a series of 100 cases pertaining to the E.N.T. domain is reported. The Entomological aspects were studied in 20 cases. The clinical features of myiasis have been described. This disease commonly occurs in diseased nasal and ear cavities which allow maggots to grow. The use of maggot oil drops and irrigation with normal saline provides an effective treatment. Prevention of maggot infestation is stressed.


The oto-palato-digital syndrome.

The oto-palato-digital syndrome. A boy aged ten years with oto-palato-digital syndrome is discussed. Because of severe conductive hearing loss tympanotomy was performed and abnormal poorly mobile ossicles were found. Stapedectomy was performed without improvement of hearing.


Wound infection following laryngectomy. The effect of topical ampicillin and carbenicillin. A study of twenty-two consecutive laryngectomies performed in one unit is reported. In eleven controls, seven (67%) developed a wound infection. In eleven patients, IG of Ampicillin and IG of Carbenicillin were used topically, and three (18%) developed a wound infection (P less than 0-04). Studies of the bacterial flora at the time of operation suggest that the major source of wound contamination is colonization of the upper gastrointestinal and respiratory tract. The ease with which a small innoculum of pathogens produces wound infection in these patients is discussed, and it is suggested that the underlying defect is depression of host cellular immunity resulting in failure of bacterial destruction by activated macrophages.


Deep vein thrombosis in ear, nose and throat surgery. I. One hundred and three patients undergoing head and neck or ear operations were screened for deep vein thrombosis using I125 labelled fibrinogen. 2. No thrombosis was detected in thirty-five patients undergoing ear operations. 3. Nine (16%) of fifty-six patients who underwent head and neck operations subsequently developed deep vein thrombosis. In a further twelve patients undergoing head and neck operations the ankle was used as a drip site and seven of these patients developed thrombosis in that leg.


Cochlear implants.

Cochlear implants. All presently devised single channel devices generate a primitive sensation of hearing by the mechanism of 'periodicity pitch'. No 'place pitch' encoding is possible. Although some enhancement of communicative skills with lip reading results, unaided speech discrimination is not possible. Definite psychological advantages for the totally deaf have been observed with these simple devices. Multiple segments of auditory nerve must be stimulated in a manner which will simulate the complex patterns of neural activity necessary for speech discrimination. Electrode optimization and the pathophysiological consequences of electrical stimulation of the auditory nerve can best be determined in animals. The perceptual consequences of electrical stimulation of the auditory nerve, however, can best be determined in man. How much we will have to innovate the methods of aural rehabilitation will depend upon how well we can generate perceptual speech patterns by electrical excitation of the auditory nerve.


The cochlear histopathology of chronic intracochlear implantation. A fundamental question regarding the feasibility of artificial electrical stimulation of auditory nerve in cases of sensory deafness is the fate of that nerve after electrode implantation. The results of chronic scala tympani electrode implantation in normal and neomycin-deafened cats indicate that the vast majority of primary auditory neurons will survive the initial surgical implantation and the long-term interface with the molded silastic electrode for periods of at least 30 months (the longest normal-implant survivors). While there is histologic evidence that some neurons are lost in the basal coil, especially in neomycin-deafened animals, the majority of spiral ganglion cells and their radial fibres survived neomycin administration and surgical implantation even in this region. The implantation of electrodes in neomycin-deafened cats did not result in heavy neuronal degeneration. Indeed, there was little apparent difference in nerve survival between the implanted and unimplanted ears of the neomycin animals for periods up to five months (the longest neomycin implant survivor). Traumatic electrode insertion with injury to the boney covering of the modiolus or disruption of the basilar membrane resulted in extensive nerve loss in the traumatized region. When carefully inserted by an experienced otologist, the molded-silastic scala tympani electrode permitted discrete differential stimulation of restricted segments of auditory nerve and produced little or no neural degeneration.


Secretory otitis. Histopathology and goblet-cell density in the Eustachian tube and middle ear in children. The histological changes in the middle ear of four children with incipient or mild chronic secretory otitis are described. They consist in vascular dilatation and proliferation, round-cell infiltration, epithelial metaplasia into pseudostratified, columnar, ciliated epithelium posteriorly in the middle ear, formation of abnormal mucous tubular glands, and an increase in goblet-cell density in the osseous tube and middle ear. The causes of these changes were presumably long-lasting tubal occlusion due to a permanent nasogastric tube and protracted catarrhal diseases. The density of goblet cells was determined in various parts of the Eustachian tube and middle ear. The findings were analysed statistically and compared with the density in prematures and newborns, normal children, and normal adults. In the tympanic orifice and in the osseous Eustachian tube the goblet-cell density was greatly increased, whereas the increase in the middle ear varied individually, but was in conformity with the other findings. These cases illustrate that the histopathological changes in the middle-ear mucosa in secretory otitis must be regarded not only from a qualitative, but certainly also from a quantitative point of view.


The aetiology of otitis externa--a new concept. This paper reports the results of middle ear pressure measurements on 33 patients with recurrent otitis externa and compares the results with those of 21 normal subjects. The maximum negative pressure in the control group was -50 mm. of water, while 17 patients (52%) with recurrent otitis externa recorded measurements of -100 mm. of water or more. It is postulated that chronic Eustachian tube dysfunction may interfere with the normal migration of epithelium from the tympanic membrane along the external auditory meatus leading to irritation and chronic infection. The treatment in these cases should be directed at the underlying cause of the Eustachian tube dysfunction.


Analysis of bile acids in conventional and germfree rats. The well-known bile acid analysis technique used by us and others (Grundy, Ahrens, and Miettinen. 1965. J. Lipid Res. 6:397-410) does not allow for the detection of hyodeoxycholic acid, a product of quantitative importance in rodent feces. Using updated methodology, it was established that hyodeoxycholic acid and omega-muricholic acid, both apparent conversion products of beta-muricholic acid, occur in apppreciable amounts in intestinal contents and feces of conventional Wistar type Lobund rats. In conventional rats, these bile acids comprise about 50% of fecal bile acids; they are not found in intestinal contents or feces of germfree rats. Others have demonstrated that hyodeoxycholic acid if formed by combined action of gut flora and liver. A new method for the separation of conjugated and free bile acids in biological samples was developed. Results with this method confirmed the total conjugation of bile acids in the germfree rat, and the almost total deconjugation that takes place in the cecum of the conventional rat.


Preparation of laurylsarcosyltaurine: a surface active constituent of crab gastric juice. The conditions for the preparation of laurylsarcosyltaurine from lauric acid and sarcosyltaurine, a crystalline dipeptide, are described. The preparation of sarcosyltaurine from the reaction of taurine and the mixed anhydride of benzyloxycarbonylsarcosine and isobutyl chloroformate is also described.


Halofenate and clofibrate: mechanism of hypotriglyceridemic action in the rat. Rats fed a fat-free diet containing no drug, 0.02% or 0.10% halofenate, or 0.25% clofibrate for 14 days were injected intravenously with equivalent amounts of either [2-3H]glycerol or [1(3)-3H]glycerol. Blood samples were collected at times up to 150 min after injection and serum triglycerides were isolated and assayed for radioactivity. Kinetic analysis of the serum appearance and clearance curves of 3H-labeled triglyceride permits estimation of serum total 3H-labeled triglyceride formation and triglyceride fractional turnover rates. The total amounts of 3H-labeled triglyceride formed from [2-3H] or from [1(3)-3H] glycerol in control-fed animals were very similar. Over 95% of the serum 3H-labeled triglyceride formed from either substrate circulated in a rapidly turning-over triglyceride pool (t1/2 = 8 min). Treatment with 0.10% halofenate or 0.25% clofibrate decreased labeling of serum triglycerides by 75-80% without increasing serum 3H-labeled triglyceride fractional turnover rates. Furthermore, both drugs decreased incorporation in vivo of 14C from [U-14C]glycerol into hepatic but not intestinal triglycerides without significantly decreasing incorporation of 14C into total phospholipids of either tissue. From these observations we suggest that, in the intact normal rat, sustained reduction of serum triglyceride levels produced by treatment with halofenate or clofibrate is due to inhibition of hepatic triglyceride formation.


Characterization of myelin of chick sciatic nerve during development. Myelin was isolated from the sciatic nerves of chicks of ages 18-day embryonic, 1-day, 4-day, 7-day post-hatch, and adult to study developmental changes in lipid composition of this structure. The yield of myelin increased throughout the early stages of development and the preparations were of high purity. Although the lipid content of the myelin did not change, significant changes took place in lipid composition during development. The most significant changes were a relative increase in cerebrosides, phosphatidalethanolamine and long-chain fatty acids of cerebrosides, and a relative decrease in the content of phosphatidylserine and phosphatidylethanolamine. A second fraction ("lower band") was obtained during the isolation procedure. This "lower band" was present at all developmental stages and layered consistently at the interface of 1.2 and 0.8 M sucrose on a discontinuous gradient. The quantity of this fraction did not change during development and it differed from myelin in electron microscopic appearance. Its lipid composition, which did not change, resembled that of 18-day embryonic myelin in its high phospholipid:cholesterol ratio and low galactolipid content. The enzyme, 2':3'-cyclic-nucleosidemonophosphate phosphodiesterase was found to be present in both the myelin and "lower band" fractions; however there was no enrichment of this enzyme in purified myelin.


An enzymatic method for the determination of the initial rate of cholesterol esterification in human plasma. A method is described for the determination of the initial rate of cholesterol esterification in human plasma, based on the enzymatic determination of free cholesterol in the plasma before and after incubation at 37 degrees C. The cholesterol esterification rate was linear up to 40 minutes. In 18 normal male and 10 normal female subjects the cholesterol esterification rate was 91 +/- 15 (mean +/- SD) and 62 +/- 12 nmoles/hr/ml of plasma, respectively.


Automated glass capillary gas-liquid chromatography of fatty acid methyl esters with reference to cis and trans isomers. The availability of an excellent separation method for fatty acid methyl esters, including separation of cis and trans isomers and of isomers that differ only in the position of double bonds, has become more and more important. The present glass capillary chromatography system combines high separation power with high precision and easy handling. Moreover, the system is completely automated and therefore provides a time saving method. As compared to a conventional packed column, the glass capillary column provides about one hundred fold more theoretical plates (227,000), as well as narrower peaks, thus giving rise to less error when integrating with electronic integrators. The reproducibility for relative retention time is better with the capillary column (0.26%) and reproducibility of the weight percent values is at least similar to that of the packed column (1.53%). When handling only small sample amounts the capillary provides better values because of its low capacity. This powerful system should open up new possibilities in the field of fatty acid investigation.


Human fat cell sizing--a quick, simple method. Two methods were used to determine the mean cell diameters of 37 samples of human adipose tissue, obtained by open or needle biopsy. Method I was the sizing of cells in cell suspensions and Method II was a quick, simple method of sizing cells from fixed sections. The agreement between the two methods was good (r = 0.93, P = less than 0.001). The results using Method II were slightly lower than those using Method I, and a correction factor is suggested. Method II has several advantages over Method I and we propose that it is a suitable method for sizing cells when a quick method with a permanent record is required.


Sterol balance in hyperlipidemic patients after dietary exchange of carbohydrate for fat. Dextrose was exchanged isocalorically for polyunsaturated fat in the liquid formula diets of 10 hyperlipidemic patients maintained under metabolic steady state conditions. Carbohydrate caused an increase of plasma triglycerides in all 10; plasma cholesterol rose in 7, and 6 of these 7 failed to show any increase in total fecal excretion of cholesterol. In contrast, fecal steroid excretion increased significantly in the three patients who maintained an unchanged or lower plasma cholesterol on the high-carbohydrate diet. Squalene, an obligatory precursor in the biosynthesis of cholesterol, rose in the plasma during carbohydrate feeding in 6 out of 6 patients studied. After a single intravenous infusion of radioactive cholesterol, plasma and feces were analyzed for specific activity over at least a four-month period. Plasma and fecal neutral sterol specific activity were essentially equivalent at all times in all patients, regardless of feeding regimen. On institution of carbohydrate feeding, the slope of cholesterol specific activity flattened for those seven patients who had a rise in plasma cholesterol concentration. There was no change in slope for the two patients with fixed plasma cholesterol levels nor for the one patient with a decreased plasma cholesterol on the high-carbohydrate diet. These experiments demonstrate a divergent response between plasma cholesterol concentration and cholesterol excretion. To establish causal relationship between the two (i.e., plasma cholesterol increases because excretion does not increase) will require further research. The flattening of specific activity die-away curves with rising cholesterol concentrations is best explained by mobilization of slowly turning-over tissue cholesterol into plasma. A subsequent decrease in cholesterol synthesis would also add to the slower decline of plasma cholesterol specific activity.


Oestriol, oestradiol-17beta and the proliferation and death of uterine cells. It has been suggested that oestriol protects against breast cancer, because in some experiments on uterine growth it is only weakly active, and partially inhibits the effects of oestradiol-17beta. When its effects are measured 24 h after a single injection, oestriol behaves as a typical impeded oestrogen with low potency and a flat dose-response line. This does not result from failure to stimulate certain critical stages of growth but from failure to sustain the products of growth. We found that oestriol induced all phases of uterine growth including DNA synthesis and cell division. It was as effective as oestradiol in stimulating early increases in protein synthesis and uterine weight, and half as effective in stimulating epithelial cells to replicate DNA and divide. However, epithelial cell numbers did not increase after a single injection of oestriol because cell death rate increased at the same time as mitotic rate, apparently as a result of the more rapid loss of oestriol from the uterus. Repeated injections of oestriol prevented premature cell death and produced as much uterine hypertrophy and hyperplasia as oestradiol-17beta. These results support the thesis that the oestrogenic potency of a substance is largely determined by the duration of its occupation of receptors. Thus in situations of continuous production, (e.g. pregnancy) oestriol would be as active as oestradiol and unlikely to exert any significant 'buffering' or protective action. The findings are also discussed in relation to a new model for the regulation of cell proliferation.


Effects of new multi-site hormone blockers on the fertility of male rats. The effects on the fertility of adult male rats of six new synthetic steroids: I, 3-cyano-5alpha-androst-1-en-17-one; II, the 17beta-acetate form of I; III, 17beta-hydroxy-5beta-cyano-androstan-3-one; IV, 6-methylpregnenolone; V, 17beta-hydroxy-17alpha-ethynyl-5beta-cyano-19-norandrostan-3-one; and VI, 19-norspiroxenone (oestr-4-en-3-one-spiro-17alpha-2'-[tetrahydrofuran]) have been tested. After 6 weeks of treatment with daily doses of 5 mg (I, II, III), 15 mg (IV) or 10 mg (V, VI) only steroid VI blocked the completion of spermatogenesis and reduced the number of foetuses sired in at least five females/male. Steroid VI also diminished seminal vesicular, prostatic, testicular and epididymal weights. It inhibited the testicular enzymes, 3beta-hydroxysteroid dehydrogenase-delta4-5-3-oxosteroid isomerase system, 17alpha-hydroxylase, and C17-20 lyase markedly, but did not affect the adrenal dehydrogenase-isomerase system. It depressed, strikingly, testicular and serum levels of testosterone and 5alpha-dihydrotestosterone and reduced pituitary and serum levels of FSH and LH. Although marked depression of target organ weights also occurred with steroids II, IV and V, and reduction of androgen levels and LH in the circulation with III, IV and V, only VI was a potent blocker of male fertility with the exception of a slight block of the siring of viable foetuses by steroids IV and V. The major difference in site of action of steroid VI from the others was the depression of pituitary and serum levels of FSH along with a marked diminution of testicular content of both testosterone and 5alpha-dihydrotestosterone. 19-Norspiroxenone in the rat is a potent anti-oestrogen without inherent oestrogenicity and is anti-uterotrophic. Thus, VI may affect male fertility by virtue of its potent anti-oestrogenic action in the hypothalamus or testis.


Plasma follicle-stimulating hormone during photoperiodically induced sexual maturation in male Japanese quail. Follicle-stimulating hormone (FSH) was measured in Japanese quail using a heterologous radioimmunoassay, the specificty of which was confirmed by its cross-reactions with purified chicken FSH and luteinizing hormone (LH). Plasma concentrations of FSH, LH and testosterone were determined in quail during the testicular growth and sexual maturation which follows their transfer from short to long daylengths. All three hormones could be detected in short-day birds but their concentrations were greatly increased following photostimulation. Plasma FSH increased 12-fold during the first 9 long days, remained at this level for a week, and then declined steadily so that by the time the birds were sexually mature the level of FSH had decreased to one-third of the maximum level. LH reached a high level (five times the short day level) after 4 long days. Thereafter two patterns of LH secretion could be distinguished. In one experiment the high level of LH was maintained unchanged throughout sexual maturation while in another experiment LH secretion decreased significantly between days 11 and 28 of photostimulation. A strong correlation existed between testicular growth and the plasma FSH concentration. It was maximal during the phase of rapid testicular growth and decreased as spermiogenesis began. The pituitary FSH content increased during photostimulation. Castration caused a 20-fold rise in plasma FSH compared with that in intact quail. The change in LH concentration after castration was about eightfold. The changes in hormone secretion were strikingly similar to those found during sexual development and puberty in the rat.


Ovine placental lactogen: acute effects on intermediary metabolism in pregnant and non-pregnant sheep. The intravenous administration of ovine placental lactogen to pregnant and non-pregnant sheep produced significant acute decreases in plasma free fatty acid, glucose and amino nitrogen concentrations. Plasma insulin concentrations decreased 1 h after administration of ovine placental lactogen and then increased significantly above baseline concentrations. The results suggest that, like human placental lactogen, ovine placental lactogen is important in the modulation of intermediary metabolism during pregnancy. The sheep is an excellent animal model for the investigation of the physiology of placental lactogen.


Observations on the localization of recently synthesized catecholamines in chromaffin cells after the injection of L-(2,5,6-3H)dopa. The fate of tl-[2, 5, 6-3h] dopa, and the intracellular localization of its metabolic products dopamine, noradrenaline and adrenaline, have been determined by the simultaneous use of assay techniques following separation of amines by chromatography and light and electron microscopic autoradiography. During the first 24 h after i.v. or i.p. injection of [3H] DOPA, synthesis of the above catecholamines occurred. Throughout this time the labelled amines were associated with chromaffin granules or immediately adjacent cytosol and not with either the Golgi complex or rough endoplasmic reticulum. Labelling of chromaffin granules occurred simultaneously throughout the cell and there was no evidence of regions containing recently labelled granules and others containing previously charged (older) granules. Adrenaline-storing cells took up [3H] DOPA and its products more rapidly and lost recently synthesized adrenaline more rapidly than noradrenaline-storing cells took up and stored their equivalent amines. This was in keeping with a more rapid turnover of catecholamines in adrenaline-storing elements.


Experimental diabetes insipidus in a marsupial, Macropus eugenii (Desmarest). A new method is described for placing lesions in the supraoptico-neurohypophysial tract to produce diabetes insipidus. The method is particularly useful with species such as Macropus eugenii in which different individuals show a large variation in relative skull measurements. The development of the diabetes insipidus syndrome in the tammar wallaby (Macropus eugenii) is shown to have essentially the same triphasic pattern of urine production as in eutherians. Increased values of adrenal corticosteroids (less than 16 mug/100 ml plasma) were observed in the oliguric interphase. This has not been previously reported and we suggest that these increased values are most probably due to potentiation of corticotrophic releasing factor by the uncontrolled release of vasopressin during this phase.


Immunoreactive alpha- and beta-subunits of luteinizing hormone in human peripheral blood and follicular fluid throughout the menstrual cycle, and their effect on the secretion rate of progesterone by human granulosa cells in tissue culture. The concentration of the common alpha-subunit of the glycoprotein hormones and of the beta-subunit of luteinizing hormone (LHbeta) in peripheral blood and follicular fluid was measured throughout the menstrual cycle, and the effects of these subunits, either alone or in combination, on the production of progesterone by human granulosa cells in tissue culture were investigated. Changes in the serum concentration of alpha-subunit and immunoreactive 'LHbeta-like' material throughout the menstrual cycle were similar to those of LH. The concentrations of the subunits before the mid-cycle gonadotrophin peak were not significantly different from those during the luteal phase of the menstrual cycle. Gel filtration of a pooled serum sample obtained at mid-cycle confirmed the presence of immunoreactive alpha-subunit together with intact LH; however, because of the cross-reactivity of LH in the LHbeta assay a distinct peak of LHbeta-subunit could not be demonstrated. In follicular fluid, alpha-subunit was detectable in large follicles (greater than or equal to 8 mm) throughout the menstrual cycle at concentrations similar to those found in serum. By contrast alpha-subunit in small follicles (less than 8mm) and 'LHbeta-like' material in all follicles were only detectable during or just after peak concentrations in peripheral plasma. The LH subunits did not increase the rate of progesterone secretion by human granulosa cells when each was added alone, even at concentrations five times higher than those in plasma. However, when both subunits were added simultaneously there was an increased rate of progesterone secretion comparable to that achieved with intact LH. It is concluded that the common alpha-subunit circulates in blood independently of the intact hormones, and that it is present in a proportion of developine Graafian follicles without affecting either the viability or biosynthetic potential of their granulosa cells. During the late follicular phase however, when both the alpha- and LHbeta-like subunits are present in follicular fluid, they may recombine and enhance steroid production by granulosa cells which are undergoing luteinization at this time.