Partial trisomy 10p and familial translocation t(7;10)(p22;p12). A girl with partial trisomy for the short arm of chromosome 10(p12leads to pter) due to mal chromosome segregation in the father 46,XY,t(7;10)(p22;p12) is described. The major abnormalities in this case are: mottled skin, mid-facial hypoplasia, low percentiles for weight, length, and head circumference, and club feet.


A  chiasma map of man.

A  chiasma map of man. By fitting compounds beta distributions to chiasma frequencies the physical map obtained from banded chromosomes has been converted into a chiasma map giving the distribution of observed chiasmata in relation to several hundred cytological bands, assuming proportionality of mitotic and meiotic chromosomes. This is a genetic map if there is a precise correspondence between sites of chiasmata and crossing-over. However, if there is appreciable preanaphase movement of chasmata, then the chiasma map is a serious distortion of the genetic map. Predictions from the chiasma map can be confirmed or refuted only by genetic evidence for which the estimates of this paper serve as initial values to begin maximum likelihood iteration.


Origin of human trisomics and polyploids. A mathematical theory is derived for the distribution of heteromorphisms in trisomics and polyploids. Applied to the sparse literature, it appears that most trisomy 21 is due to first division maternal nondisjunction, although there are case reports of second division nondisjunction, both paternal and maternal. Most triploids appear to be due to dispermy or failure of the first meiotic division in the oocyte. The need for larger systematic samples is stressed. This maximum likelihood analysis gives great resolution to epidemiological studies, permitting discrimination of etiological factors acting on fathers and mothers, at different meiotic stages, and on dispermy.


Further studies on the biological properties of guinea-pig IgG1 antibodies. III. Haemolytic efficiency in vitro. The capacity of non-reciprocally contaminated guinea-pig IgG1 and IgG2 antibodies for sensitizing antigen-coated sheep erythrocytes for lysis was studied in the presence of serial dilutions of normal guinea-pig serum as the source of complement. IgG1 antibodies were highly efficient, provided complement was little diluted and not heated at 56 degrees. Chelation of Ca2+ by EGTA affected lysis of IgG1-sensitized cells by only 50%, while completely blocking lysis of IgG2-sensitized cells. The F(ab')2 fragment of IgG1, in contrast to the F(ab')2 fragment of IgG2, was almost as efficient as the parent antibody.


The effect of immunological adjuvants on the relative affinity of anti-protein antibodies. Inbred mice of a strain (B1OD2 new) known to produce either no detectable antibody or antibody of low affinity to two protein antigens administered in saline, were immunized with human serum transferrin (HST) in one of nine adjuvants. Such immunization increases the level and relative affinity of anti-HST antibody. The adjuvants used varied in the degree to which they augmented these parameters of the antibody response--that is, FCA and FIA were capable of inducing high levels of high affinity antibody, whereas other adjuvants elicited lower levels of high affinity antibody. The possibility is discussed that substances with adjuvant activity may effect antibody production at two stages: (1) at the stage of antigen selection of cells for proliferation and (2) at the stage or proliferation of antibody producing cell precursors.


The response of protein-deficient mice to tetanus toxoid. Effects of antigen dose, adjuvants, period of deprivation and age on antibody production. The effects of a 4% albumin diet initiated at weaning on primary and secondary responses to AIPO4-adsorbed and soluble tetanus toxoid (TT) were studied in C57BL mice. Responses of both groups were directly proportional to the dose of antigen over most of the range tested, but at very low doses protein-deficient mice produced higher primary titres than normal mice. In the primary response AIPO4-adsorption of the antigen essentially increased the effective dose irrespective of the diet, but after secondary challenge responses to soluble TT were more severely affected by the diet. Normal secondary titres to AIPO4-adsorbed TT were achieved when deficient mice were given high doses of antigen. Diet also affected the relative proportions of IgG and IgM produced in most responses. Gram-negative bacterial vaccines and lipopolysaccharide increased antibody production in both groups of mice. The low protein diet produced less dramatic effects when initiated at the time of inoculation or later, and mice maintained for longer on the diet produced more nearly normal titres. Mechanisms which may explain these findings are discussed.


Passive cutaneous anaphylaxis and vasoactive amine challenge in nude (nu/nu) mice. A comparison with nu/+, axenic and young normal animals. Recent suggestions of a thymic origin or thymo-dependent differentiation of tissue mast cells prompted an investigation in the athymic nude (nu/nu) mouse. The outbred nu/nu examined were found to possess mast cells in at least comparable numbers to the phenotypically normal nu/+. These nu/nu were superior to nu/+ as recipient for mast cell-dependent, long latent period (IgE-type), passive cutaneous anaphylactic (PCA) reactions. A variety of studies performed, including competition with nu/+ serum, thymic restoration, quantitation of nu/+ homocytotropic antibody responses, comparison with axenic or young mice and time-course studies of skin sensitization suggested that the higher PCA titres in nu/nu skin were not entirely due to a lack of competitive inhibition of mast-cell sensitization. The nu/+ used in these studies were in fact poor IgE recipients, but were somewhat more sensitive than nu/nu to hypovolaemic shock induced by histamine and serotonin mixtures. Pretreatment with Bordetella pertussis greatly enhanced the sensitivity of nu/+, axenic, holoxenic and young normal mice to the vasoactive effects of histamine and serotonin, but was somewhat less effective in nu/nu and old mice.


Enthalpy-entropy compensation in dinitrophenyl--anti-dinitrophenyl antibody interaction(s). The effect of varying the temperature over a wide range (-3 dergees -67 degrees) on the binding of xi-DNP-L-Lysine to bovine colostral anti-DNA IgG1, and also rabbit anti-DNP IgG revealed non-linear van't Hoff plots. The extent of the curvatures were found to be indicative of large positive heat capacity changes; and the thermodynamic parameters, calculated using a non-linear least-squares computer procedure for these anti-DNP antibody preparations, revealed an enthalpy-entropy compensation mechanism for hapten-antibody binding. The enthalpy factor was found to be the primary contributor for the binding process at low temperatures, but at increasing temperatures the entropy factor assumed greater importance. At physiological temperature (37 degrees), the entropy factor was the major contributor to the free energy of reaction for rabbit anti-DNP IgG, while for bovine colostral anti-DNP IgG it was predominant at temperatures higher than 37 degrees.


Induction of acute and chronic arthritis by intra-articular injection of preformed collagen-anticollagen complexes. Three groups of rabbits were injected intra-articularly into the knee joints with preformed complexes of hydroxamated collagen and antiserum to denatured collagen respectively, with hydroxamated collagen and normal serum, hydroxamated collagen, anticollagen serum or saline as controls. From twelve rabbits, receiving only one injection and investigated 8 h later, only the joint tissues of rabbits which received immune complexes showed acute arthritis with severe infiltration of polymorphonuclears, leucocyte thrombi in vessels and haemorrhages. From six rabbits, receiving two consecutive injections (day 0 and day 2) and investigated 12 h after the last injection, only the joint tissue of rabbits which received immune complexes showed subacute-chronic arthritis. From sixteen rabbits receiving four consecutive injections day 0, day 2, day 7 and day 14) and investigated 7 days after the last injection, only the joint tissue of rabbits which received immune complexes showed severe chronic arthritis with infiltration of plasma cells, lymphocytes and nodule-like accumulation of lymphocytes whilst their joint fluid showed additionally a distinctly increased number of polymorphonuclears (3-15 x 10(6)) which differed with P less than 0-01 statistically from all controls.


Leucocyte chemotactic activity in the parallel bioassay of guinea--pig lymphokines. This paper describes the application of parallel bioassay to determine the extent to which the leucocyte chemotactic activity of guinea-pig lymphokine preparations is associated with their lymphocyte mitogenic, inflammatory (increased vascular permeability) and macrophage migration inhibitory activities. A convenient test of chemotactic activity was devised whereby Coulter counter determinations were made of the number of peritoneal exudate cells passing through a Nuclepore membrane into the medium of the lower (test) compartment of a chemotaxis chamber. Symmetrical parallel line (2+2) dose assays were used to obtain potency ratio estimates of the chemotactic activity of seven lymphokine preparations whose mitogenic, inflammatory and migration inhibitory activities were already known. Statistical analysis of the results revealed a clear dissociation of chemotactic activity from mitogenic and migration inhibitory activities; but with six of the seven lymphokine preparations, a marked similarity was revealed between their chemotactic and inflammatory activities. Cytological studies showed that although the lymphokine preparations were chemotactic for different cell types, there was a preferential migration of neutrophils and the smaller mononuclear cells. The demonstration by parallel bioassay of an association between the ability of lymphokine preparations to increase vascular permeability in vivo and to promote chemotaxis of leucocytes in vitro is viewed in relation to the possible role of lymphokine in inflammatory processes.


Functional affinity constants of the reaction between 125I-labelled C1q and C1q binders and their use in the measurement of plasma C1q concentrations. Purified 125I-labelled C1q has been found to react with glutaraldehyde-treated red cells (glut-RBC), with a value for the functional affinity constant, K, of 1-3 X 10(8) M-1, based on measurement of concentrations of bound and free reactants at equilibrium. Values of K obtained for other C1q-binders were as follows: diaminopropane, 2 X 10(2) M-1; monomer IgG, 5 X 10(4) M-1; heat-aggregated IgG, 0-5-2-5 X 10(8) M-1; IgG-anti-IgG complexes, 0-31 X 10(8) M-1. The functional rate constant for association (ka) between 125I-labelled C1q and glut-RBC was 5 X 10(5) M-1 S-1 at 37 degrees in 0-17 M NaC1. The rate of dissociation of the C1q-glut-RBC complex was biphasic with rate constants (kd) of 2 X 10(2) S-1 and 2 X 10(5) S-1. Calculated values of K from the ration ka/kd gave values of 2-5 X 10(7) M-1 and 2-5 X 10(10) M-1. It is suggested that the range of values of K reflects the involvement of 1,2 or more binding sites on the C1q molecule. Reduction of the ionic strength of the medium from 0-17 M to 0-14 M increases the rate of association of C1q and glut-RBC eleven-fold, indicating involvement of ionized groups at the binding site. A method is described for measuring plasma C1q concentrations by saturation assay, using 125I-labelled C1q and glut-RBC. Plasma C1q concentrations fell in the range 170-250 microng/ml.


Endothelial adenine uptake as an assay for cell- or complement-mediated cytotoxicity. [3H] Adenine uptake by monolayers of cultured pig aortic endothelium was characterized and used as the basis for a terminal adenine uptake cytotoxicity assay. This assay was used to demonstrate specific cytotoxicity against donor-type endothelium by serum and by peripheral blood lymphocytes from skin-graft allergized pigs, and to compare the time course of these two types of effector mechanism.


Rat transplantation antigens. V. A variation of the KCl-method releasing low molecular weight antigen. Extraction of soluble histocompatibility antigen from normal rat lymphoid cells by hypertonic salt (3 M KC1) releases high mol. wt material. It the original lymphoid cell suspension is washed more than once before extraction, antigenic acitivity disappears as measured by inhibition of cytotoxic antibody. This activity is mostly retained in the supernatants of the washing procedures. Extraction of these cell-free supernatants produced an antigenic material with a mol. wt of approximately 60,000. This suggests that at least part of the material accessible for KCl-extraction is released very early from the cell surface from the mechanical action, necessary for preparing the cell suspension.


Antibody-independent activation of the alternative complement pathway in human serum by parasitic cells. The lysis of the monocellular parasite Trypanosoma cyclops by normal human serum (NHS) was found to be complement-dependent and to follow activation of the alternative pathway without apparent requirement for conventional antibodies. Lysis of the organisms was inhibited by heat-inactivating NHS at 56 degrees, preincubation of NHS with cobra venom factor or chelation of divalent cations with EDTA. It took place, however, in human C2-deficient serum and was inhibited by prior heating of NHS at 52 degrees to destroy the activity of factor B of the alternative pathway. Moreover, the lytic reaction was magnesium- but not calcium-dependent. Repeated low-temperature (0 degrees) absorption of either human hypogammaglobulinaemic serum or NHS with the parasite failed to remove or significantly decrease their lytic activities.


Influence of aromatic compounds on the interaction of activated C4 with EAC1. The influence of amino acids and their derivatives on the formation of SAC14 from SAC1 and C4 were investigated. Among the compounds tested, aromatic amino acids containing phenolic hydroxyl group or carbobenzoxyl group inhibited the formation of SAC14. When aromatic compounds other than amino acids were tested, the inhibitory capacities of these compounds were found to relate to the substituent group incorporated in the aromatic ring. Aromatic compounds with subsituent groups with negative Hammett's substituent constant showed stronger inhibition. Conversely, compounds with group with positive constant showed weaker inhibition. These results may suggest that the increased electron density in the aromatic ring is responsible for the inhibition. Regardless of inhibition of SAC14 formation, these inhibitors did not affect the enzymatic action of EAC1 or C1 on C4 but inhibited the binding of activated C4 to EAC1.


Host-parasite relationships in the yeastlike form of Paracoccidioides brasiliensis strain IVIC Pb9. The yeastlike form of Paracoccidioides brasiliensis strain IVIC Pb9 reduced the amount of alpha-1,3-glucan in its cell wall from 45 to 3% when subcultured in vitro for several years. This strain regained its alpha-1,3-glucan up to 25% of the total cell wall when grown in vivo. A mutant strain of P. brasiliensis Pb9, named IVIC Pb140, reported to have 1,3-mannan instead of alpha-glucan in the cell wall, could not be recovered from experimentally infected animals. The existence of some relationship between the presence of alpha-1,3-glucan in the cell wall of the yeastlike form and the pathogenicity of this fungus is suggested in this report.


Selection of influenza B virus recombinants and their testing in humans for attenuation and immunogenicity. The selection of influenza B virus recombinants from plaques in bovine kidney cell monolayers is described. Two sets of recombinants were each derived from parents of high and low virulence for humans, respectively. Recombination frequency was apparently high, and reassortment of genes made it possible to obtain attenuated recombinants containing the surface antigens of the virulent parents. Attenuation and immunogenicity were demonstrated in a series of volunteer trials. However, technique proved less satisfactory than for influenza A viruses which periodically undergo antigenic shift and for which there is a wide choice of parent viruses with distinctive surface antigens. In our two influenza B recombinant series there was appreciable antigenic overlap in the neuraminidases of the parents, even though in both cases these were chronologically widely separated. Another marker used was comparative titer at 35 and 38 degrees C. In practice, technical problems might sometimes make it difficult to ensure rapid production of live influenza B vaccines by recombination.


Immunoglobulin and specific-antibody synthesis in vitro by enteral and nonenteral lymphoid tissues after subcutaneous cholera immunization. An in vitro culture technique with synthesis of 14C-labeled protein has been used to study immunoglobulin and specific-antibody formation by spleen, mesenteric lymph nodes, Peyer's patches, and small intestine of rabbits, which were immunized twice subcutaneously with Vibrio cholerae lipopolysaccharide (LPS) and enterotoxin; saline-injected rabbits served as controls. Newly synthesized immunoglobulin G (IgG), IgA, and IgM were quantitated by liquid scintillation after their isolation by means of affinity chromatography from columns with immunoglobulin class-specific antibodies coupled to Sepharose. Specific antibodies were similarly measured after purification from gels derivatized with LPS or cholera toxin. The isolated antibodies had full biological activity as studied in protection tests. The immunization increased the overall IgM synthesis in the spleen. It also enhanced the production of IgA and IgG in Peyer's patches and of IgA in intestine. Significant synthesis of radiolabeled antibodies against both V. cholerae LPS and enterotoxin was found in spleen as well as in Peyer's patches of immunized animals. Titration with an enzyme-linked immunosorbent assay (ELISA) showed significant levels of IgG as well as IgA antibodies in incubation medium from all the studied tissues, whereas specific IgM was only found for spleen and mesenteric lymph nodes. Simultaneous tissue incubations at 37 degrees C and in an ice bath indicated that the major part of the antibodies registered with the ELISA represented de novo synthesis.


Brain lysosomal glycosidase activity in immunosuppressed mice infected with avirulent Semliki forest virus. Mice infected with an avirulent strain of Semliki forest virus show an increase in the activity of some of the brain lysosomal glycosidases. The increase in activity of these enzymes has been correlated with the histological, virological, and serological changes that result from the infection in the presence and absence of immunosuppression. Semliki forest virus alone caused the development of a mild encephalitis with perivascular infiltration, microgliosis, astrocyte hypertrophy, and a focal spongiform encephalopathy, together with an increased activity of brain N-acetyl-beta-D-glucosaminidase and beta-glucuronidase. Antilymphocyte serum given after infection marginally affected the course of the disease. Cyclophosphamide markedly delayed the development of the spongy changes in the increase in enzyme activities, but not the perivascular infiltration. It is suggested that the increased activity of the lysosomal glycosidases studied may be linked both to the development of a successful immune response and to the focal spongiform changes produced by the infection.


Virulence of Streptococcus mutans: cariogenicity of S. mutans in adult gnotobiotic rats. Gnotobiotic rats infected with Streptococcus mutant 6715, mutans C211 at 45 days of age on provided a purified diet containing 5% sucrose developed carious lesions on buccal, sulcal, and proximal molar surfaces within 15 days (60 days of age). The level of caries increased significantly (P less than or equal to 0.01) within the next 15 days (by day 75), an extensive decay was observed on all three molar surfaces of 90-day-old infected rats (45 days after challenge). Mutant C211 was previously shown to exhibit increased glucosyltransferase activity and greater adherence and virulence than S. mutans 6715 wild type (wt). Gnotobiotic rats (90 days of age) infected with either S. mutans AHT or S. mutans 6715 (wt) at 45 days of age developed significantly (P less than or equal to 0.01) fewer caries on all molar surfaces than rats of the same age that were infected with S. mutans 6715, mutant C211. The level of plaque increased 2-fold, and the number of viable S. mutans in plaque increased 10-fold between days 60 and 90 in rats infected with S. mutans 6715, mutant C211. Ninety-day-old rats infected with either S. mutans AHT or S. mutans 6715 (wt) had similar levels of plaque and numbers of S. mutans in plaque; however, these values were two- to fourfold lower than those observed in rats of the same age that were infected with S. mutans 6715, mutant C211.


Release of beta-lysin from platelets caused by antigen-antibody complexes, purified enzymes, and platelet-aggregating substances. The release of beta-lysin, which followed the intravenous injection of antigen-antibody complexes, did not take place when these complexes were added to citrated whole blood but did occur in heparinized blood. beta-Lysin release in heparinized blood was inhibited by citrate but were reversed by the addition of calcium ions that implicated complement reactions. Fourteen different enzymes were added to platelet-rich plasma (PRP). Streptokinase, neuraminidase, papain, phospholipase C, sulfatase, and trypsin caused platelets to release significant quantities of beta-lysin, whereas elastase, phosphatase, protease, ribonuclease A, hyaluronidase, lipase, and pepsin caused little or no increase in the plasma beta-lysin concentration. One enzyme, fibrinolysin, inactivated beta-lysin faster than it was released. The enzyme-induced release of beta-lysin from PRP was often accompanied by a reduction in the number of platelets. The intravenous injection of streptokinase, neuraminidase, and sulfatase caused in vivo releases of beta-lysin into the plasma. The platelet-aggregating substances collagen, arachidonic acid, and adenosine 5'-diphosphate caused beta-lysin to be released from PRP. The platelet-aggregating substances L-epinephrine, zymosan, fibrinogen, reserpine, and serotonin caused little or no release of beta-lysin from platelets. The results of this study indicate that the release of beta-lysin during antigen-antibody-complement reactions, blood coagulation, phagocytosis, and inflammation could be enzyme mediated.


Evaluation of surface components of Vibrio cholerae as protective immunogens. Surface components of a motile Inaba strain (CA401) were removed from washed cells by low-speed shearing. Flagella contaminated with a vesicular material (designated as crude flagella [CFA1) were obtained by differential centrifugation of the shear fluid. Vesicles were obtained from a nonflagellated mutant by the same procedure. Homogeneous small vesicles were obtained in diminished yield from CsCl gradients of CF preparations. Treatment of CF with sodium deoxycholate removed the vesicular material and flagellar sheaths and yielded naked flagella (NF). The ability of these preparations of passively protect infant mice suckled by CFW mothers that had been immunized at the time of mating was compared, on a dry-weight basis, with commercial vaccine (CV). Eight-day-old mice were challenged orally with more than 1,000 50% lethal doses of either the homologous or a heterologous (Ogawa Ca411) strain. The most effective immunogen was CF, which provided complete protection at 1 microng against both challenges. CF and vesicles provided 50- to 100-fold greater protection than CV against homologous challenge. With heterologous challenge, vesicles were 10-fold more protective than CV, markedly less protective than CF. The NF offered only slightly greater protection than CV against both challenges. Immunoelectrophoresis revealed an antigen in CF distinct from vesicles, cell wall lipopolysaccharide or NF. This antigen is not present in the nonflagellated mutant and is apparently associated with motility,


Fat of Coxiella burnetti in macrophages from immune guinea pigs. The interaction between Coxiella burnetii and peritoneal macrophages obtained from immune guinea pigs was studied by transmission electron microscopy. Phagocytosis and subsequent fate of ingested phase I and II rickettsiae were compared. Phase I rickettsiae were more resistant to phagocytosis than were phase II organisms. Macrophages from phase I- and II-immunized animals were equally capable of phagocytizing rickettsiae. Phase I and II rickettsiae previously treated with normal serum multiplied and destroyed macrophages from guinea pigs that had been immunized with phase II rickettsiae. Phase II organisms were initially suppressed in macrophages from phase I-immunized animals, but eventually multiplied in these cells. In contrast, only phase I organisms were destroyed by macrophages from phase I-immunized animals. Treatment of rickettsiae with immune serum enhanced ingestion by macrophages and potentiated the destruction of organisms by both types of macrophages. The macrophage migration inhibition assay was performed on peritoneal exudate cells from immune animals. Migration of peritoneal macrophages from phase I-immunized guinea pigs was inhibited, whereas macrophages from phase II-immunized animals migrated when cells were cultured in the presence of killed, intact phase I or II C. burnetii.


Novel bacterial structures in human blood: cultural isolation. Evidence for the existence of a novel bacteriological system has been obtained from osmotically lysed and filtered human blood (membrane filters with a pore size of 0.22 micronm) placed in special culture media. These blood filtrates gave rise to ordinary bacteria for 71% of the blood specimens processed from diseased humans and for 7% of those from supposedly normal humans. Morphologically, the bacteria resembled streptococcal, staphylococcal, and gram-positive filamentous (cocco-bacillary) forms. Prior to the appearance of bacteria in the media, large and small "dense bodies" were microscopically observed but disappeared when ordinary bacteria were apparent, Cultures of of unlysed blood as conventionally performed were negative. These organisms may represent an adaptation of certain bacteria to life in the blood.


Enteropathogenicity of various isolates of Treponema hyodysenteriae. Isolates of Treponema hyodysenteriae from 25 geographically separated outbreaks of swine dysentery were tested for their ability to produce the disease. Clinical signs and lesions typical of acute swine dysentery were produced in 52 of 68 (75%) susceptible specific pathogen-free pigs that had been orally inoculated with pure cultures of 23 of 25 beta-hemolytic isolates. In addition, 13 weakly beta-hemolytic isolates of nondysentery origin with morphology similar to T. hyodysenteriae did not produce disease when orally inoculated into susceptible specific pathogen-free pigs. Two of these latter isolates, Puppy and B296, and one pathogenic, beta-hemolytic isolate failed to produce disease when orally inoculated into puppies.


Cancer in the Africans and Arabs of Zanzibar. Among 392 cancers histologically diagnosed in Zanzibar during 1957-62 and 1964-67, a definite pattern seems to emerge. Skin and cervix cancers were the most common types in both Africans and Arabs. Skin cancer was predominantly of the squamous cell type. The Zanzibar Arabsthus appear to be protected against basal cell carcinoma which in the Arab desert community has been diagnosed with about the same frequency as squamous-cell carcinoma. In the same manner, the Zanzibar Arab immigrants seem to have a reduced risk for stomach and oesophageal cancers, which are common in other Arab countries. This is probably because Arabs in Zanzibar have adopted the dietary habits and other customs of the Zanzibar Africans in whom cancer of the alimentary tract seems to be uncommon. On the other hand, unlike Zanzibar Africans, the Arabs have an increased risk for Hodgkin's disease similar to that of the Middle East Arab population.


A 51chromium isotope release assay for detecting cytotoxicity to human bladder carcinoma. A 51chromium isotope release assay is described for measuring in vitro cytotoxicity reactions against allogenetic target cells. Lymphoid cells from some patients with localized transitional cell carcinoma (TCC) were shown to selectively lyse targets derived from two long-term cell lines of TCC. The effector cells in this reaction did not form E rosettes. Cytotoxicity was successfully recovered after cryopreservation.


Detection of cellular immunity to tumor-associated antigen(s) in mice by the macrophage spreading inhibition test. Soluble preparations of antigens from a methylcholanthrene-induced fibrosarcoma of C57BI mice were prepared by homogenization of tumor tissue and high-speed centrifugation of the homogenate. These preparations were able to sensitize syngeneic mice to tumor-associated antigens (TAA) of the fibrosarcoma, and to provoke a delayed hypersensitivity reaction when injected into the footpad of sensitized mice. Furthermore, the same soluble preparations could inhibit in vitro the spreading of peritoneal macrophages from mice sensitized to TAA. A similar inhibition of macrophage spreading was obtained when peritoneal cells from C57BI mice, bearing transplants of the fibrosarcoma, were incubated with the preparations. We conclude that the macrophage spreading inhibition test, like other in vitro assays, can detect cell-mediated immunity to tumor-associated antigens.


Metabolism of the carcinogenic hydrocarbon benzo(a)pyrene in human fibroblast and epithelial cells. II. Differences in metabolism to water-soluble products and aryl hydrocarbon hydroxylase activity. Aryl hydrocarbon (benzo(a)pyrene) hydroxylase (AHH) activity and metabolism of benzo(a)pyrene to water-soluble products were measured in cultures of body fibroblasts and kidney epithelial cells from different human embryos. AHH activity at 24 h after treatment with or without benz(a)anthracene was determined in cultures from 23 embryos, and 3 days' accumulated metabolism of benzo(a)pyrene to water soluble products was measured in cultures from 18 embryos. The body fibroblasts from the different embryos could be divided into three groups according to the amount of water-soluble products, but not according to the AHH activity. These three groups were not found by either assay in the cultures of kidney epithelial cells. In both fibroblast and epithelial cells, high metabolism to water-soluble products was not necessarily associated with high AHH activity. The results extend our previous finding (Huberman and Sachs, 1973) of three presumably genetic groups for BP metabolism to water-soluble products in human fibroblast but not in epithelial cells and indicate that this grouping was not found in these cells by measuring AHH activity.


HLA in familial Hodgkin's disease. Results and a new hypothesis;. A familial aggregate of seven cases of Hodgkin's disease (HD) has been investigated by HLA typing. Over 600 people in the immediate population (i.e. about half) have been HLA typed and haplotypes have been obtained for 95% of them. It was expected that the cases would share a particular HLA haplotype or at least that they would have one or two HLA antigens of the same series in common. However, this was not the case so no simple idea of association of HLA with HD cases was upheld. When antigen frequencies were examined in the whole population, it was found that HLA B18 increased progressively in incidence from 0.08 to 0.4 in successive groups of individuals each one more closely related to the HD cases. Similarly the community with the highest incidence of HD also had the highest incidence of B18. Thus B18, which in the world figures carries the highest relative risk, emerged as important in this study. Of four proposed interpretations of the data, we are most interested in the idea that the important HLA association is at a population level rather than at the level of the individual patient. A hypothesis, based on the concept of a "healthy carrier" for the HD agent, explains how such an association might operate. It is possible that B18-linked complement deficiency could be the basis for such a carrier state.


Chromosome 14 translocation in African and North American Burkitt's lymphoma;. Two established North American Burkitt lymphoma cell lines were studied by chromosomal banding techniques. The SU-AmB-1 line previously shown to be negative for the Epstein-Barr virus (EBV) was found to have, among other changes, a translocation from the long arm (q) of chromosome 8 onto 14q. The SU-AmB-2 line, which contains the EBV genome, also displayed the same 8/14 translocation. These results were compared with data from three EBV-positive tumor cell lines derived from patients with African Burkitt's lymphoma. Our findings indicate that a translocation from 8q onto 14q occurs in both African and North American Burkitt lymphomas, and that this abnormality apparently is not related directly to EBV. This chromosome translocation therefore may be an important event in the development of human lymphocytic malignancy, analogous to the occurrence of the Philadelphia chromosome rearrangement in chronic myelogenous leukemia.


Relationship of human natural lymphocyte-mediated cytotoxicity to cytotoxicity of breast-cancer-derived target cells. Mononuclear cells from 115 individuals were tested in a 4-h chromium release assay against two breast-cancer-derived cell lines, G11 and MCF-7, and a myeloid line, K-562, shown previously to be sensitive to natural cytotoxicity. These data were analyzed in a manner designed to detect hyperreactivity against the breast cell lines relative to the level of reactivity against K-562. A high proportion of breast cancer patients were found to be relatively hyperreactive against G11 (12/18 or 67%) and against MCF-7 (10/18 or 56%). Fibroadenoma patients were very similar to the normal females, with 0/11 hyperreactive to G11 and 1/11 (9%) to MCF-7. However, several normal males (7/17 or 41%) were hyperreactive to G11 but not to MCF-7 (2/17 or 12%). Colon cancer and lung cancer patients were also more hyperreactive to G11, 4/8 or 50% and 4/6 or 67%, respectively, than they were to MCF-7, 1/8 or 13% and 1/6 or 17%, respectively. Only fibrocystic patients resembled the breast cancer patients, with some but not as many individuals being hyperreactive to G11 (3/8 or 38%) and to MCF-7 (2/8 or 25%). With another group of individuals reproducibility of the method was demonstrated, with only 1/14 or 7% of normal females and 12/17 or 70% of breast cancer patients being hyperreactive to G11. Thus, natural cytotoxicity toward K-562 can be related to breast cancer-associated cytotoxicity toward MCF-7 in a way that distinguishes a majority of breast cancer patients specifically from other groups of individuals.


"Sternberg-reed" giant cells of Hodgkin's Disease: cultivation in vitro, heterotransplantation, and characterization as neoplastic macrophages. Cells from the involved spleens of 25 patients with Hodgkin's disease have been grown in long-term culture and compared with normal spleen macrophage cultures from control cases. The Hodgkin's spleen cell culture contained mono-, bi-, and multi-nucleate giant cells, many closely resembling Sternberg-Reed cells, which were adherent, phagocytically active and neoplastic by the dual criteria of aneuploidy and heterotransplantability. Lysozyme secretion was consistently observed in all Hodgkin's cultures tested. The giant cells possessed both Fc and complement (c3b) receptors, and lacked lymphocyte markers such as (c3b) receptors, surface IgM, and the capacity to form E-rosettes. Binucleate and multinucleate cells, as well as mononuclears, were capable of active DNA synthesis, and binuclear mitotic figures were observed. It is concluded that these cells are the in vitro descendants of the Sternberg-Reed and Hodgkin neoplastic cell population, and that they are derived from macrophages or closely related cells of the mononuclear phagocyte system.


Origin of embryo-derived yolk sac carcinomas. The origin of yolk sac carcinoma obtained from rat embryos transplanted to extrauterine sites was traced to the extraembryonic portion of 9-day egg-cylinders. Under appropriate conditions cells of the extraembryonic portion of the egg cylinder differentiate into cells of parietal yolk sac epithelium, continue to proliferate and form retransplantable malignant tumors. Serum concentrations of alpha-fetoprotein were elevated in rats bearing yolk sac carcinomas and in some animals bearing teratomas admixed with yolk sac carcinoma. Possible factors that regulate the survival and proliferation of yolk sac epithelium in extrauterine sites are briefly discussed.


Induction by cigarette smoke of aryl hydrocarbon hydroxylase activity in the rat kidney and lung. In both the rat kidney and lung, inhalation of cigarette smoke diluted with air (1/15) for a limited period of time (15 min) specifically induces aryl hydrocarbon hydroxylase (AHH)2 in less than 4 h. Up to four successive inhalations administered at 2-h intervals activities. The maximal effect corresponds to about 10 times the control value. Compared to the kidney enzyme, the lung AHH activity is about three or four time more sensitive to small concentrations of cigarette smoke. The biological half-life of the lung AHH activity is longer than 24 h, while it is only 3-4 h in the kidney. In both tissues, the induced enzyme presents the same in vitro thermolability and sensitivity to various inhibitors. For the establishment of the AHH induction, protein synthesis is continuously required, while RNA synthesis is only necessary during the first 2 h following the smoke treatment.


Augmentation of natural cytotoxic reactivity of mouse lymphoid cells against syngeneic and allogeneic target cells. The levels of natural cell-mediated cytotoxicity against tumor cells in young BALB/c and BALB/c nude mice could be augmented by inoculation of a variety of mouse tumor cells or of mouse thymocytes. In older mice with low levels of spontaneous cytotoxic reactivity, inoculation of tumor cells led to rapid appearance of cytotoxicity. This augmented cytotoxicity reached a peak 3 days after inoculation and then declined rapidly. The specificity of the augmented cytotoxicity appeared to be the same as that seen with natural cell-mediated cytotoxicity. The detected antigens were restricted to mouse tumor cells and thymocytes, and were absent on cells from other species. The effector cells after boosting also had the same cell surface characteristics as the natural cytotoxic effector cells, being non-adherent, non-phagocytic, and only weakly sensitive to treatment with anti-theta serum plus complement. In addition to this boosting by mouse tumor cells, marked increases in the levels of cytotoxicity were caused by a variety of murine viruses, including murine sarcoma virus and lymphocytic choriomeningitis virus. These effector cells also had the same properties as those seen with natural cytotoxic effector cells. The results indicate that the levels of natural cell-mediated cytotoxicity in conventional and athymic BALB/c mice can be consistently and rapidly boosted by inoculation with tumor cells or viruses. This should provide a valuable tool for better understanding of the mechanisms responsible for the expression of natural cytotoxicity and its relevance to in vivo resistance to tumor growth.


Tumor cell locomotion--a factor in metastasis formation? Influence of cytochalasin B on a tumor dissemination pattern. The influence of Cytochalasin B (CB) on TA3 ascites tumor cells was studied in vivo in order to assess whether CB-induced cell paralysis would affect the transplanation behavior of the cells and in particular tumor distribution after IV cell infusion ("experimental metastases"). Tumor cell pre-treatment with CB (1 mug/ml) did not alter the SC or IV transplantability of TA3 cells. Pre-treatment with 10 mug/ml CB, in contrast, consistently increased the incidence and number of extra-pulmonary tumor takes from IV transfused cells. The amount of pulmonary tumors was not significantly altered. SC transplantability was not affected by 10 mug/ml CB. The importance of cell mobility and cell surface topography for tumor cell nidation in vessels is discussed.


Effects of sex hormones on oncogenesis in rat urinary bladder by N-butyl-N-(4-hydroxybutyl)-nitrosamine. The effects of testosterone propionate, estradiol, and estriol on the oncogenesis of male rats' bladder induced by oral administration of the carcinogen N-butyl-N-(4-hydroxybutyl)-nitrosamine (BBN) were examined. There were significant differences between the testosterone propionate-administered and estradiol-administered groups and also between the testosterone propionate-administered and estriol-administered groups in the incidence of bladder tumors.


Blood and saliva salicylate measurement in the monitoring of salicylate therapy. Total and free plasma salicylate as well as saliva salicylate was measured in 13 patients with rheumatoid arthritis treated with different doses of acetylsalicylic acid alone. Although the clinical response was judged moderately effective to effective in all cases, total and free plasma salicylate varied markedly. Saliva salicylate correlated well with both total plasma salicylate and fre plasma salicylate. Determination of saliva salicylate may be a simple and useful technique for the monitoring of salicylate therapy.


Gentamicin pharmacokinetics during hemodialysis in patients suffering from chronic renal failure. Because the elimination of gentamicin, a potent aminoglycoside antibiotic, is dependent almost entirely on renal excretion, renal functional impairment drastically changes the pharmacokinetics of this drug. As a first step in the study of the effects of renal insufficiency and the anephric state on the pharmacokinetic parameters of gentamicin, serum and urine levels of this drug were studied after a single intravenous bolus dose during hemodialysis in patients suffering from chronic renal failure. The data were fitted to the two-compartment open model and the appropriate kinetic parameters were calculated with the COMPT computer program modified by Pfeffer. The rate constant of metabolism was estimated from plasma and dialysis rate constants of elimination. The rate of renal excretion was shown to be very weak in patients who were not anuric. The use of the mathematical equations of the two-compartment open model demonstrated that, after a single dose of gentamicin, the percentage of decrease of serum concentration with time does not represent, because of tissue binding retention, the percentage of drug eliminated from the body. It was shown that pharmacokinetic parameters represent a useful tool in the optimization of gentamicin therapy.


Computer-calculated kanamycin dosage regimen and monitoring. The applicability of a computerized dosage regimen determination based on a recently developed method for drugs following the minimum inhibitory concentration pattern (bacteriostatic drugs) has been tested for kanamycin injected intramuscularly in 8 patients with varying degrees of renal impairment. The dosage regimens for maintaining a minimum therapeutic kanamycin concentration of 5.0 micron g/ml was determined with the Wang 700 C Computer. After the 1st, 2nd, 3rd, 6th, 9th, and 12th dosing interval blood samples were taken and the acual kanamycin serum concentrations determined. The expected multiple dose serum level curves for each patient were simulated using the Comdyna Dose Generator Analog Computer and the individual actual serum concentrations were monitored. The actually found serum level data were compared to the expected values based on the dosage regimen equations and to the steady state equations. Additionally, the expected peak maxima were calculated. The proposed method resulted in effective and safe kanamycin serum levels.


The effect of pyridinolcarbamate after acute and chronic administration in patients with atherosclerosis obliterans. The study was aimed at evaluating the effect of pyridinolcarbamate in patients with atherosclerosis obliterans of the lower limbs. A single intravenous administration of the drug did not induce changes in resting blood flow, but elicited a significant decline in capillary filtration rate. In a double-blind cross-over trial patients reported a significant prolongation of walking distance after 6 months of pyridinolcarbamate treatment. Sensation of cold in the lower limbs was not affected. Pyridinolcarbamate did not influence peak values of blood flow through the calf but they appeared significantly earlier. No significant changes were seen at the evaluation of the elevation and dependency test and performance using tiptoeing test. Likewise the findings of systolic pressure in the lower limbs, pulse wave morphology, and claudication distance on treadmill did not confirm a therapeutic effect of pyridinolcarbamate.


Effect of xanthinol nicotinate treatment on platelet aggregation. Treatment of 24 male patients with 3 g/day of xanthinol nicotinate did not change the in vitro measurements of ADP-induced platelet aggregation but produced a marked inhibition of collagen-induced platelet aggregation. This effect may be connected with the drug-induced depression of the ATP level in platelet-rich plasma. Changes in the platelets in the patients' blood or in the lipid composition and the concentration of uric acid in their serum were ruled out as reasons for the decrease of the collagen-induced aggregation. The activity of the three serum enzymes y-GT, GOT, and GPT and the concentration of the blood sugar did not change.


Fast reliable photography for the dermatologist. Color photography using a 35 mm single lens reflex camera is a useful tool for both academic and practicing dermatologists. With an electronic flash and a macro lens, the production of high quality photographs requires only a few seconds. Equipment and techniques effective in office photography are surveyed and evaluated.


Solid phase synthesis and some hormonal activities of 8-L-homoarginine-vasotocin and 1-deamino-8-L-homoarginine-vasotocin. 8-L-Homoarginine-vasotocin and its 1-deamino derivative were synthesized by the solid phase method. Both compounds possessed high activities in the uterus contraction and fowl depressor assays and showed higher pressor activities than the corresponding vasopressins. Homoarginine-vasotocin, unexpectedly, was a more potent antidiuretic agent than its deamino analogue.


Preliminary X-ray crystallographic and physico-chemical investigations of human ceruloplasmin. Single crystals of the plasma protein ceruloplasmin (CP) and its two modified forms: neuraminidase-treated CP (asialoCP) and NaN3-inhibited CP (NaN3-CP) suitable for X-ray studies have been grown. The native CP crystallizes as described previously by Magdoff-Fairchield et al. (1969) in the tetragonal space group 14 (a = b = 268.2 A, c = 129.1 A) with two protein molecules in the asymmetric part of a unit cell. AsialoCP crystals belong to the trigonal space group P 3(1)21 or P321 (a = b = 215.0 A, c = 84.5 A) and have one protein molecule in the asymmetric part of a unit cell. NaN3-CP crystals are isomorphous to crystals of native CP. Despite some differences in electrophoretic mobility and optical properties, the conformations of the native CP molecule and its modified forms are similar, as can be concluded from a study of ORD and CD spectra.


Investigation of the role of tryptophan in alpha-MSH. Replacement by L-pentamethylphenylalanine and L-phenylalanine. Two analogues of alpha-MSH are described, in which the tryptophan residue occuring in position 9 of the natural hormone has been replaced by pentamethylphenylalanine and phenylalanine, respectively. The analogues were synthesized via a conventional procedure and the [Phe9]-analogue also by a semi-synthetic approach, which demonstrated the favourable properties of the applied, new amino-protecting Msc function for this purpose. The widely different electron donor properties of the substituted residues were accompanied by a large difference in melanocyte stimulating activity of the analogues. The [Pmp9]-analogue, having donor properties comparable to those of the natural compound, was four to five times more active than the analogue containing the poorly donating Phe residue. The opposite effect was noted in in vivo lipolysis in rabbits.


The influence of esterification of carboxyl groups of ribonuclease-A on its structure and immunological activity. The effect of modification of carboxyl groups of Ribonuclease-Aa on the enzymatic activity and the antigenic structure of the protein has been studied. Modification of four of the eleven free carboxyl groups of the protein by esterification in anhydrous methanol/0.1 M hydrochloric acid resulted in nearly 80% loss in enzymatic activity but had very little influence on the antigenic structure of the protein. Further increases in the modification of the carboxyl groups caused a progressive loss in immunological activity, and the fully methylated RNase A exhibited nearly 30% immunological activity. Concomitant with this change in the antigenic structure of the protein, the ability of the molecule to complement with RNase-S-protein increased, clearly indicating the unfolding of the peptide "tail" from the residues for orthobenzoquinone reaction and the loss in immunological activity of the more etion of these derivatives as compared with the compact native conformation. The fact that even the fully methylated RNase-A retains nearly 30% of its immunological activity suggested that the modified protein contained antibody recognizable residual native structure, which presumably accommodates some antigenic determinants.


A comparison of the amino-terminal sequences of two calf-thymus chromatin non-histone proteins. The N-terminal sequences of two calf-thymus non-histone chromosomal proteins, HMG-1 and HMG-2, are compared. Both N-terminal sequences are basic, and differ at only one position. This contrasts with the C-terminal portion of HMG-1 which we have previously shown to be high in acidic residues. There is no obvious sequence homology with any of the known sequences of the histones.


Stereochemical restrictions on the occurrence of amino acid residues in the collagen structure. The primary structure of collagen is characterized by the repeating tripeptide sequence (Gly-R2-R3)n. The results of theoretical studies, carried out using contact criteria to compute the stereochemically allowed orientations for various side chains at locations 2 and 3, are reported here. It is found that side chains with only gamma-atoms, as in valine, serine and threonine, or with only one delta-methyl group, as in isoleucine, can occur equally well at locations 2 and 3, as is actually the case in collagen. Side chains with two Cdelta-atoms, as in leucine and phenylalanine, can also be accommodated at both positions. However, if they occur as R3, their freedom of orientation is severely restricted in the presence of a proline residue as R2 in a neighbouring chain. If water molecules bound to the chains of the triple helix are assumed to be present, then location 3 is virtually impossible for leucine and phenylalanine residues. Location 2 is, however, unaffected, and their presence as R2 can help to shield the water molecules from disturbance by the solvent medium. This may be the reason for the preferential occurrence of Leu and Phe residues in location 2 in the collagen triplets, although the polypeptides (Gly-Pro-Leu)n and (Gly-Pro-Phe)n form collagen-like structures.


Studies of the affinity of human serum albumin for binding of bilirubin at different temperatures and ionic strength. The association constants for the binding of bilirubin to human serum albumin (HSA) have been determined at four different temperatures by measurements of the rate of the peroxidase catalyzed oxidation of unbound bilirubin. The change of enthalpy is determined from a van't Hoff plot (ln Kass versus 1/T) to about -13.5 kcal/mol. deltaG degrees is calculated from the binding constants, and deltaS degrees is obtained from: deltaG degrees = deltaH degrees--TdeltaS degrees. The results show that the large negative deltaG degrees (--11 kcal/mol) for binding of bilirubin to HSA is a consequence of the negative deltaH degrees. The entropy was found to be about--8.5 cal/mol/degree and tends to diminish the numerical value of deltaG degrees. The binding constant has also been determined at varying ionic strength. The results show a decrease in binding for increasing salt concentration. The data from the two sets of experiments suggest that hydrogen bonds and salt linkages rather than hydrophobic interactions are the main factor in the binding of bilirubin to its primary site on HSA.


Thiamine deficiency in fulminant hepatic failure and effects of supplementation. Nine out of 24 patients with acute hepatocellular necrosis leading to fulminant hepatic failure showed biochemical evidence of thiamine deficiency early in the course of their illness, probably as a result of inadequate intake of the vitamin. This was corrected by twice daily administration of intravenous vitamin supplements containing thiamine hydrochloride (100 mg b.d.). These studies indicate that conversion of thiamine hydrochloride to its biologically active co-enzyme form, thiamine pyrophosphate, is possible even in the presence of severe acute hepatocellular necrosis, and it is suggested that supplements of the vitamin should be included in the routine management of patients with acute hepatic failure.


[Niacin efficiency in zein base diets].

[Niacin efficiency in zein base diets]. Lots of growing or adult rats consume ad libitum diets based on zein (12 p. 100 of protein) (diets A), or on zein partially supplemented by lysine and tryptophan (= protein quality of maize) (diets B), or on zein supplemented by amino acids to fill its deficiencies totally (diets C). The diets contain 15 mg of niacine per kg. The diets A 1, B 1 and C 1 receive an addition of 10 mg of supplementary niacin. The efficiency is estimated by the amount of niacin and tryptophan in blood plasma and in liver. The niacin efficiency depends more on the protein quality - more precisely of the intake of tryptophan - than the intake of niacin: the hepatic concentrations of niacin are 120 mug with diet A and 210 mug with diet C in growing rats. These variations are identical to the evolution of intakes and of concentrations of liver nitrogen. The administration of a supplement of niacin (10 mg/kg) does not cause repercussion on the body niacine on the growing rat and provokes a very weak profit on the adult animal. In the dietsA (unsupplemented zein), only the tryptophan is an efficacious source of niacin. The niacin seems unable to increase the vitamin storage in the liver.


Effect of dietary vitamin E on lipid synthesis by rat lung in vitro. The effect of dietary vitamin E on lipid synthesis from U-14 C-D-glucose and 1-14C-acetate was studied in rat lungs in vitro. One-month-old Sprague-Dawley male rats were fed either a basal vitamin E-deficient diet or one supplemented with 45 ppm vitamin E ad libitum for two months. Glucose oxidation to CO2 by lungs was significantly (p less than 0.05) decreased by the exclusion of vitamin E from the diet. Oxidation of acetate to CO2 was not affected by the presence of vitamin E in the diet. The extent of labeled carbons from both glucose and acetate incorporated into total lipids was significantly lower in the lungs of vitamin E-deficient animals than in those of the supplemented group. However, the relative amounts of phospholipids, neutral lipids are free fatty acids in total lipids, and of glyceryl moiety and fatty acids in total lipids and in phospholipid fraction were not significantly altered by the status of dietary vitamin E. The results suggest a general depression of lipid synthesis in the lungs of vitamin E-deficient rats.


[Efficiency of various B vitamins in zein base diets]. Lots of growing rats consume ad libitum rations based on zein (diets A), or zein partially supplemented in lysine and tryptophan (= protein quality of maize) (diets B), or zein supplemented in amino acids to fill totally its deficiencies (diets C). The diets contain 15 mg of niacin, 1.5 mg of thiamine, 2.5 mg of riboflavin and 1.5 mg of pyridoxine per kg. The efficiency of these vitamins is estimated according to their hepatic amounts. Compared with the diet A, the liver of animals of diet B contains in average 37 p. 100 less vitamins and that of diet C 56 P. 100 less. The amount of niacin in liver directly depends on the tryptophan intake and not on that of the niacin. The amount of riboflavin and pyridoxine depends on the nitrogen in the liver: the retention of 1 mg of nitrogen in the liver provokes the retention of 1.1 mug of riboflavin and of 0.2 mug of pyridoxine. The amount of thiamine is less narrowly linked to the hepatic nitrogen metabolism. The constitution of vitamin storage in the liver comes from the protein quality of the diet and from the possibilities of nitrogen storage.


The validity of biochemical assessment of thiamine, riboflavin and folacin nutriture. Nutriture of thiamine, riboflavin and folacin was assessed by two tests: thiamine (109 subjects) by the TPP effect of erythrocyte transketolase activity and urinary excretion of thiamine (mug/g creatinine); riboflavin (81 subjects) by the activation coefficient of erythrocyte glutathione reductase and excretion of riboflavin in urine (mug/g creatinine); and folacin (91 subjects) by estimation of folacin in red blood cells and in serum (ng/ml). The following correlation coefficients (r) were obtained: transketolase activity vs thiamine excretion: -0.33; glutathione reductase vs riboflavin excretion: -33; and red blood cell folacin vs serum folacin: 0.77. When "deficient" and "low" values were defined as "not acceptable" and compared with "acceptable" values, sensitivity of thiamine excretion was 54%, of riboflavin excretion 33% and of serum folacin level 90%. The respective value of specificity were 75%, 83% and 37%. Sensitivity of thiamine excretion and of serum folacin level, respectively, increased when stricter criteria of insufficiency were applied and assessment of "deficiency" of these vitamins rather than of "non-acceptability" was attempted.


The effect of riboflavin deficiency on white cell glutathione reductase in rats. 1. The measurement of glutathione reductase in white cells from peripheral rat blood is described; the contribution from red cell glutathione reductase was virtually eliminated. 2. The activation of the white cell enzyme by flavin adenine dinucleotide (FAD) in vitro was measured during an eight week period of dietary riboflavin deficiency in weanling rats. 3. The activation increased progressively during the deficiency reaching about fifty per cent after eight weeks, at which time liver FAD levels had declined to 30% of the control levels and clinical symptoms of deficiency were beginning to appear. No increase in activation of white cell glutathione reductase was observed in pair-fed controls, over this time period. 4. These observations suggest that the measurement of the activation coefficient of white cell glutathione reductase could usefully supplement that of the red cell enzyme, in defining riboflavin status.


Effect of graded biotin levels in the diet on liver pyruvate carboxylase of chicks fed ad libitum and after starvation. Decreasing levels of biotin in the diet from 200 mug/kg to 0 mug/kg reduced the pyruvate carboxylase activity per g liver and in the whole liver of chicks to 17% of the normal activity. Oral supplementation with 3 mg biotin per chick increased the enzyme activity within 24 hours back to normal. In animals well supplemented with biotin (200 mug/kg diet) 24 hours of starvation increased the per g liver pyruvate carboxylase activity. When the pyruvate carboxylase activities were related to the whole liver, no effect of starvation was found.


L-ascorbate 2-sulfate and mobilization of cholesterol from plaque deposited in rabbit aortas. Rabbits were randomly assigned to two groups. Age and body weight distribution were equal in both groups. All animals were placed on a high cholesterol diet for 9 wk, then returned to a normal diet for 1 wk. At the end of this dietary regimen, one group of animals received subcutaneous injections of psysiological saline 3 times/day, 5 days/wk for 10 wk, and the other group recieved L-ascorbate 2-sulfate (0.37 mmole) according to the same timetable. On alternate weeks, the serum levels of total and free cholesterol were determined. After 10 wk of treatment the animals were killed; the plaques were excised from the aortas and examined for total mass and cholesterol content. We observed that, under these dietary conditions, L-ascorbate 2-sulfate does not mobilize cholesterol or its esters from preformed aortic plaque. However, we did observe that animals showing high cholesterol levels in their sera died prematurely when injected with L-ascorbate 2-sulfate. Gross and histopathological investigations of organs and tissues did not reveal any significant differences among the animals that died prematurely and those that survived to the termination of the experiment.


Blood levels and bioavailability of ascorbic acid after administration of a sustained-release formulation to humans. Plasma and whole blood concentrations of ascorbate have been measured after administration of 1000 mg of the vitamin as standard or sustained-release capsule formulations to human subjects. A peak of mean whole blood and plasma concentrations of 8.5 mug/ml and 5.7 mug/ml respectively (corrected for predose concentrations of "endogenous" ascorbate) occurred at 12 h and 6 h respectively after administration of a sustained-release formulation. After administration of a standard formulation, the peak of mean whole blood and plasma concentrations of 9.4 mug/ml and 8.6 mug/ml respectively occurred at 4 h. In whole blood, mean concentrations of ascorbate declined with a half-life of about 34 h. The bioavailability of ascorbate from the sustained-release formulation was 149% and 180% of that from the standard formulation by calculation from whole blood and plasma concentration data respectively.


Gonadal and plasma testosterone and cholesterol in scorbutic guinea pigs. Testes of pre- and post-pubertal guinea pigs were examined histologically and biochemically to see if ascorbic acid deficiency brought about any changes in their structure or function. Apart from a slight but significant fall in gonadal cholesterol there was no apparent change in the histology or testosterone content in scorbutic guinea pigs. Plasma cholesterol and testosterone were also similar in the control and scorbutic animals.


Effects of chronic vanadium pentoxide administration on L-ascorbic acid metabolism in rats: influence of L-ascorbic acid supplementation. 1. Rats toxicated with vanadium pentoxide showed drastic retardation in growth rate and supplementation of L-ascorbic acid to these rats could not reverse this effect. The urinary excretion of L-ascorbic acid and D-glucuronic acid was decreased in the toxicated group of rats. 2. Considerable lowering of L-ascorbic acid content of the liver tissues of rats was observed under vanadium toxicated conditions. Supplementation of L-ascorbic acid to this group raised the tissue Vitamin C reserve considerably. 3. The normal histological patterns of the liver and kidney tissues of rats were severely disturbed under vanadium toxicated conditions. L-ascorbic acid supplementation to this group of rats showed marked signs of restoration in this respect. 4. Vanadium pentoxide treatment brought about a significant reduction in the biosynthetic capacity of L-ascorbic acid, along with an enhanced utilization of this vitamin. Subsequent supplementation of L-ascorbic acid to the toxicated group of rats was found to be effective in reversing these effects almost to the basal level.


Enzymatic assay of fructose-1,6-diphosphate for the measurement of its utilization by tissues. The enzymatic analysis of fructose-1,6-diphosphate by a coupled assay utilizing triose-phosphate isomerase, glycerol-3-phosphate dehydrogenase and aldolase was carried out in rat serum and human plasma after fructose-1,6-diphosphate administration. The rate of disappearance of fructose-1,6-diphosphate was determined and compared to the activity of aspecific phosphatase. The uptake of fructose-1,6-diphosphate by rat liver and ileum was also determined with the same method.


Peroxide formation, vitamin E and myocardial damage in the rat. Rats were fed diets containing 10% cod liver oil with or without dietary tocopherol for 16 or 32 weeks. Adipose tissue, skeletal muscle and myocardium were isolated. They were examined histologically and analyzed for autoxidation products and a number of energy metabolites. After 16 weeks small amounts of peroxides were present in adipose tissue as determined by the thiocyanate method. Ceroid pigment and slightly defective striation were observed in myocardium and to a lesser extent in skeletal muscle of the tocopherol deficient group. The ATP content was also significantly lower in this group. After 32 weeks, adipose tissue of the tocopherol deficient group contained large amounts of ceroid and had a high content of peroxides and other autoxidation products. More ceroid and a significantly higher peroxide estimate in lipid extracts were found in myocardium of the tocopherol deficient group as compared to the controls. Similar but weaker signs of peroxide occurrence and ceroid formation were obtained in skeletal muscle. The significance of the findings for myocardial function is discussed.


Acute inversion of the uterus.

Acute inversion of the uterus. Eight successive cases of acute puerperal inversion of the uterus are presented. The incidence was one in 26,364 deliveries. Fundal insertion of the placenta, adherent placenta and low parity were significant factors in our cases. Two patients were given halothane anesthesia to relax uterine retraction and cervical ring. Vaginal repositioning was successful in seven cases. One patient required abdominal repositioning. Apart from iron deficiency anemia, the postoperative period was uneventful.


Genital bleeding in women aged 50 and over. Histologic findings from 1,038 endometrial curettages performed for abnormal bleeding in patients 50 years old and over are presented. These findings were analyzed and compared according to the age of the patients. The overall malignancy rate was 22.7%. Endometrial carcinoma was the most common malignancy. The possibility of a malignant tumor increases with age. Atrophic endometrium was the cause of post-menopausal bleeding in 33.2% of the patients and adenomatous or adenocystic hyperplasia in 27.0%.


Perineovulvovaginal preparation in labor. Seventy-four patients who received antenatal care and whose deliveries were supervised in our hospital were studied. Most of them were from the low socioeconomic class. Forty patients were shaved and all patients had the same routine perineal, vulval and vaginal swabbing during the first stage of labor. Puerperal morbidity was low. Even in developing countries where patients often have poor personal hygiene, shaving the vulval, pubic and perineal hair is unnecessary for patients whose pregnancies and deliveries are supervised in hospital. We suggest that the procedure should be reserved for patients who have complications of pregnancies which may necessitate instrumental or abdominal deliveries.


Angular and interstitial pregnancy.

Angular and interstitial pregnancy. Two cases of interstitial and one of angular pregnancy have been presented. Angular pregnancy occurs in the angle of the uterine cavity, while interstitial pregnancy is a true ectopic pregnancy. The outcome may therefore be different: the first may develop or abort into the cavity, while the second will almost always rupture. Because of the rich vascularization of this area, hemorrhage is usually profuse and may be catastrophic. Preoperative diagnosis is rare, but the triad of bleeding in pregnancy, no fetal remnants on dialation and curettage and an asymmetric uterus suggest angular or interstitial pregnancy. More liberal use of laparoscopy may increase the number of cases diagnosed before severe bleeding occurs.


Cervical carcinoma in Zambia.

Cervical carcinoma in Zambia. A series of 274 cases of cervical carcinoma in Zambian Africans is presented. Cervical carcinoma formed 29.3% of all female cancers. Poorly differentiated squamous carcinoma constituted the most common histologic type. Concomitant bilharziasis was found in 24.4% of cases. In 59 cases causal factors such as coitus, early marriage, age at first pregnancy and parity were studied and discussed. No correlation was found between circumcision and cervical carcinoma.


Thyroid cancer after irradiation.

Thyroid cancer after irradiation. Nine patients with thyroid cancer who had received irradiation on the thyroid region (four for breast cancer with deep x-ray or Co60, one for a skin lesion with deep x-ray and four for hyperthyroidism with I131) have been discussed. These cases might be within the range of statistical coicidence, but the potential carcinogenic effect of irradiation could not be ruled out completely. The need for detailed examination of the thyroid gland before and for many years after irradiation and early removal of thyroid gland nodules which develop after I131 treatment are emphasized.


Immediate postoperative complications in the critically ill surgical patient. Immediate postoperative complications in 160 surgical patients were retrospectively reviewed. Complications were analyzed by system, and mortality was evaluated.


Culdotomy for female sterilization.

Culdotomy for female sterilization. Tubal occlusion through a culdotomy incision offers a method of surgical sterilization acceptable to women in some countries where abdominal sterilization is not acceptable. This technic requires skilled gynecologic surgeons, who are a limited resource in many developing countries. On the other hand, gynecologic surgeons practicing in outlying areas may find it more feasible to perform colpotomy sterilizations than laparoscopic sterilizations with the latter's requirements for specialized instrumentation and additional training. With adequate attention to operator training, patient selection, evaluation of patients for postoperative complications and appropriate care of those with such complications, culdotomy should be included among the various approaches appropriate for female sterilization.


Ovarian neoplasms and pregnancy.

Ovarian neoplasms and pregnancy. Sixteen cases of benign cystic ovarian neoplasms (13 serous cystadenomas and 3 dermoids) were discovered among 5,616 pregnancies. The only serious complication was torsion of the ovary containing the tumor in one case.


Choriocarcinoma in Jamaica.

Choriocarcinoma in Jamaica. Fifty-two cases of choriocarcinoma were recorded in the Jamaica Cancer Registry for the parishes of Kingston and St. Andrew, giving an incidence of 1:7,384 live births. This is intermediate between figures reported for Western metropolitan populations and for Far Eastern countries. Clinicopathological studies on 26 cases treated at the University Hospital in Jamaica revealed that 65% followed either normal pregnancy or abortion. Choriocarcinoma following a normal pregnancy or occurring without a previous history of pregnancy carried a poorer prognosis than those with a preceding hydatidiform mole. Many patients had primary neurological manifestations. The overall prognosis was poor due to late presentation with disseminated metastases. Of the 26 patients, 15 died of the disease within two years and only six appeared for a six-year follow-up.


Cesarean section for twin pregnancy in Ibadan, Nigeria. During an 11-year period, 23,524 deliveries were conducted at the University College Hospital, Ibadan; 4,360 (18.5%) were delivered by cesarean section, 49 of which were associated with twin pregnancy. The main indications for this operation in twin pregnancy were prolapsed umbilical cord, previous cesarean section scar and malpresentation of the second twin with the membranes already ruptured. The perinatal mortality of 18.2% was mainly attributable to prematurity. Obstetricians in this community have adopted a conservative attitude towards cesarean section because of the general aversion to this operation by our patients and the risk of subsequent rupture of the uterine scar in women who wish to deliver themselves at home in subsequent pregnancies. Cesarean section is therefore performed in twin pregnancy for the same absolute indications as for single pregnancy.


Induction of abortion and labor by extraamniotic isotonic saline, with or without addition of oxytocin, in cases of missed abortion, missed labor and antepartum fetal death. In a group of 16 women admitted for missed abortion, missed labor or antepartum fetal death, we induced abortion or labor by means of extraamniotic infusion of isotonic saline solution, using a Foley catheter inserted through the cervix. Eight patients also required parallel intravenous administration of 5-10 U of oxytocin drip. The average time from the onset of treatment until expulsion of the macerated fetus was 9.09 hours. No complications were recorded. This method is suggested in all such cases, especially when the classical methods are contraindicated.


Primary elective cesarean section in Ibadan, Nigeria. A total of 13,692 deliveries were conducted at University College Hospital, Ibadan. These included 2,764 (22%) cesarean sections, 63 (2.3%) of which were primary elective sections. The case records of 44 of these patients have been analyzed to determine the place of primary elective cesarean section in modern obstetric practice in Nigeria. No maternal deaths occurred and morbidity attributable to the operation was low and not serious. One perinatal death occurred and morbidity was low and less serious than maternal morbidity. There was no record of a uterine scar rupture occurring in subsequent pregnancies in any of these patients. Primary elective cesarean section, if used discriminately, will reduce difficult labor with the attendant risks of maternal and fetal mortality and morbidity in Nigeria and developing countries. The role which improved socioeconomic conditions and health education plays is emphasized.


Accumulation of taurine by cultured retinal pigment epithelium of the rat. Cultured rat retinal pigment epithelium actively accumulates taurine against a concentration gradient; an intracellular concentration of 12 mM is maintained at an extracellular taurine concentration of 0.12 mM. Taurine accumulation conforms to Michaelis-Menten kinetics for extracellular taurine concentrations between 0.006 and 0.05 mM; within that range the Km is 16 muM and the Vmax is 2.3 nmol./hr./mul of cells. Above an extracellular concentration of 0.5 mM the kinetics are complex. Accumulation is inhibited by reduced temperature and iodoacetate as well as by dinitrophenol in the absence of glucose. Ouabain, reduced extracellular sodium, and removal of extracellular potassium and calcium also inhibit taurine accumulation. The cells also release previously accumulated taurine at half-time of 1 to 3 hours. The rapid accumulation and release of taurine by cultured retinal pigment epithelial cells is consistent with the idea that this tissue is a site for the transport of taurine in vivo.


An interpretation of small-angle light-scattering patterns of human cornea. Small-angle light-scattering patterns of human cornea in I. and I modes were compared to theoretical scattering patterns generated in the computer on the basis of a cornea model. This model is a nonrandom assembly of optically anistropic rods. The best match of theoretical and experimental patterns yielded the following parameters describing the ultrastructure of human cornea: length of the rod (fiber)=12 mu; angle between optic and geometric axis of the rod=60 degrees; most probable orientation of the rod=150 degree; the width of the distribution function of rod orientation (omega0)=2.5; intrinsic birefringence of the rod=7 X 10(-6); form birefringence=1.3 X 10(-5).


The influence of age onthe sensitivity of the cornea,. Corneal touch threshold (CTT) was measured in 205 healthy people of different ages. It was found that CTT increased gradually throughout life, although more significantly after the fifth decade of life. The results are found to be in good accord with those of Boberg-Ans, thus refuting an earlier report asserting that corneal sensitivity (threshold-1) increased up to the fifth decade and declined thereafter,


Unilateral gonadectomy does not alter contralateral intraocular pressure in rabbit eyes. The purpose of this study was to determine whether unilateral gonadectomy caused any change in intraocular pressure. Repeated measurements in 22 male and nine female rabbits after unilateral gonadectomy were made by three different methods of determining intraocular pressure. We were unable to find a decrease in intraocular pressure on the side opposite to the side of the removed gonad, as had been previously reported. Neither were any ipsilateral changes observed. A calibration curve (rabbit) for the applanation pneumatonograph is also presented.


The electrical stimulation of the retina by indwelling electrodes. Chronically implanted intraocular electrodes were  used to stimulate visual cortical responses to current passage through the retina. Electrical current threshold for the light-adapted cat retina lies in the region of 30 to 100 X 10(-6) amps. for the conditions used. Dark adaptation caused a large threshold increase. Retinal excitability also decreased when electrical stimuli were delivered more often than one per 5 seconds. Effective charge-density threshold was moderately greater than reported for frogs and humans.


Calcification in retinoblastoma.

Calcification in retinoblastoma. Review of 40 cases of retinoblastoma revealed that histologic evidence of calcification was noted in 38 of the 40 tumors, whereas only three of 16 roentgenograms were positive for calcium. Calcificaton detected by roentgenograms and histopathologic examination correlated poorly with quantitative determination for tumor calcium. Compared to control eyes, however, eyes with retinoblastoma contain large amounts of calcium (1.2 vs. 218 mug/ml. ash). This calcification, though frequently not observed in standard roentgenograms, should be detected by the newer diagnostic modalities such as hypocycloidal polytomography, computerized transaxial tomography, ultrasonography, and radionuclide scintigraphy with technetium diphosphonate, a bone-scanning agent.


Heterotransplantation of retinoblastoma into the athymic "nude" mouse. Fresh surgical specimens of retinoblastoma were successfully transplanted into the anterior chamber of the "nude" mouse (a homozygous mutant, nu/nu, with a severe defect in cellular immunity), filling the eyes but failing to grow subcutaneously. Retinoblastoma cells from an established cell line, Y-79,1 spread from the intraocular injection site to invade the orbit, optic nerve, and brain and formed large tumors when implanted subcutaneously. Tumor cells injected into the anterior chamber of immunologically normal litter-males (heterozygotes, nu/+) survived for varying periods in the anterior chamber but showed little growth.


Pathology of the corneal endothelium.

Pathology of the corneal endothelium. Clinical specular microscopy has indicated that human cell healing occurs by spreading, there is a limited healing reserve, and premature cell loss is the equivalent of a "premature aging" that may lead to later decompensation. This instrument has been useful in studying healing and cell damage from surgery, drugs, and special procedures such as intraocular lens insertion. It pointed out extensive cell loss at the time of intraocular lens insertion, and subsequent studies have indicated that at least part of this cell loss may be due to the methacrylate surface of the lens. Laboratory studies suggest that coating that surface can prevent this component of cell loss. The magnitude of benefits to be found from such coating requires further clinical study.


Human cornea organ cultures: epithelial-endothelial interactions. Full-thickness explants of five human corneas grown in organ culture demonstrate that the migration of epithelium and endothelium is mutually limited once contact between the two cell types has been established. Two human corneas, however, failed to show this contact inhibition, and corneal epithelium surrounded completely the explant; both corneas showed cornea guttata. The implications of corneal epithelium-endothelium contact inhibition, and the failure of such inhibition, are discussed.


Effect of the ophthalmic preservative thimerosal on rabbit and human corneal endothelium. Widespread use of the mercurial-containing preservative thimerosal as an antibacterial agent in ophthalmic drugs and solutions warranted an investigation into its possible cytotoxic effects on the functional and ultrastructural integrity of the corneal endothelium. No changes in corneal thickness were observed during 5 hours' perfusion of the endothelium of rabbit and human corneas with 0.0001 and 0.0005 percent thimerosal in glutathione bicarbonate Ringer's solution (GBR). Scanning electron microscopy (SEM) and transmission electron microscopy (TEM) of the endothelium of the 0.0001 percent group revealed normal ultrastructure. SEM and TEM of the endothelium of corneas perfused with 0.0005 percent thimerosal for 5 hours revealed condensed mitochondria, cytoplasmic vacuoles, and cytoplasmic flaps at the apical end of the cellular junctions. Perfusion of higher concentrations (0.001 and 0.005 perecnt) of thimerosal in GBR resulted in increases in corneal thickness after 2 hours and irreversible ultrastructural damage to the endothelial cells by 5 hours. Corneas perfused with 0.01 and 0.1 percent thimerosal in GBR showed a rapid and immediate increase in corneal thickness and endothelial cell death and necrosis within 1 hour. It is postulated that the mercury in thimerosal becomes bound to the cell membrane protein sulfhydryl groups, causing an increase in cellular permeability; These results suggest that the prolonged exposure of the corneal endothelium to thimerosal in the accepted antimicrobial dosage of 0.005 to 0.001 percent may result in functional and structural damage to the endothelium.


The contribution of morphology to our understanding of the pathogenesis of experimentally produced corneal vascularization. In many disorders of the cornea, blood vessels invade this normally avascular tissue. Several theories have been proposed to explain the pathogenesis of corneal vascularization. Since none had received general acceptance, experimental studies of the phenomenon were begun several years ago in hamster cheek pouch chambers. These investigations which employed sequential morphologic studies during the process of corneal vascularization gave rise to a new hypothesis, namely, that corneal vascularization is usually a manifestation of the inflammatory response and mediated by leukocytes. This report briefly reviews the evolution, evidence, and current status of this theory.


Corneal radiofrequency burns: effects of prostaglandins and 45/80. Vascularization of the rat cornea was produced by means of radiofrequency burns adjacent to the limbus. Animals with and without lesions were tested for a vascular response to topical PGE1 and compound 48/80, with carbon used as an indicator. There was no response to PGE1 until 6 hours, at which time dense labeling of limbal vessels was observed. This response gradually decreased and by day 15, when labeling returned to control levels. No further permeability changes were seen for the duration of the experiment to day 90. Compound 48/80 gave four-plus labeling of the limbus and one-plus labeling of corneal vessels at all times during the experiment. The presence or absence of the thermal lesion did not change the degree of labeling. Histamine and bradykinin did not produce vascular labeling either with or without a corneal lesion.


Scanning electron microscopy of the trabecular meshwork in normal and glucomatous eyes. The ultrastructure of trabeculectomy specimens obtained from enucleated eyes was compared to that of 23 similar specimens extracted from glaucomatous eyes. It was thus possible to study the trabecular meshwork from Schwalbe's line to the iris root, the corneal endothelium, the canal of Schlemm, its inner and outer walls, and the collectors. In the glaucomatous specimens it was possible to verify, for the first time, two aspects which show clinical correlation. The first aspect is pseudoexfoliation: we find in the trabecular meshwork two types of deposits in great quantity. One is formed by a dense irregular fibrillar net, with a diameter oscillating betwwwn 600 and 830 A, surrounding the trabeculae; the others are pigment granules. Both fill the intertrabecular spaces. The first type is also observed on the corneal endothelium lining Schwalbe's ring, stressing the visibility of the intercellular spaces. The second aspect is the pectinate ligament (mesodermic remnant) in late-onset congenital glaucoma. Its pattern is characteristically different from that of the corneoscleral meshwork.


Effects of repeated microwave irradiations to the albino rabbit eye. Albino rabbits were exposed to repeated microwave irradiations once daily for a month. Clinical examination was carried out for a period up to one year. No changes occurred below 300 mW./cm.2. At and above this level posterior subcapsular iridescence and posterior cortical cataracts were produced. The histologic findings in posterior subcapsular iridescence are presented. It appears that the nonprogressive posterior cortical cataracts are a result of the temperature levels generated by the microwaves in the immediate retrolental area.


Experimental allergic optic neuritis in guinea pigs: preliminary report. An experimental model for acute allergic optic neuritis was produced in adult strain 13 guinea pigs by sensitization with isogenic spinal cord emulsion in complete Freund's adjuvant. These animals exhibited two distinct clinical patterns: (1) "retrobulbar optic neuritis," with a diminished pupillary response to light despite a normal fundus, and (2) "neuroretinitis," with a diminished pupillary response associated with hyperemia and swelling to the disc and juxtapapillary retinal edema. Histopathologic study of those animals with "retrobulbar neuritis" revealed that some had no abnormalities in the optic nerve or chiasm, but showed foci of mononuclear cell infiltration in the brain. Others had a mononuclear cell infiltration localized to the retrobulbar portion of the optic nerve and chiasm with multiple foci of axial and periaxial demyelination. Similar pathologic changes were present in the animals with "neuroretintis", but the lesions were located just behind the lamina scleralis. These animals also exhibited marked swelling of the axons at the lamina retinalis. On examination by light microscopy, the alterations in the region of optic nerve head appeared characteristic of papilledema.


Experimental retinal dysplasia due to cytosine arabinoside. Retinal dysplasia was produced in newborn rats treated postnatally with the antimitotic substance, cytosine arabinoside (ara-C). In rats examined from 6 to 60 days, there were numerous retinal rosettes surrounded by photoreceptor cells and bipolar cells containing photoreceptor cell processes, displaced nuclei, and cellular debris. Abnormal development and alignment of photoreceptor cell processes were commonly observed. Cellular degeneration was evident at all ages, and infiltrating phagocytic cells were especially numerous in the retina of treated rats examined at 60 days. Characteristic features of ara-C-induced retinal dysplasia included the scattering of bipolar cell nuclei in the inner and outer nuclear layers and marked reduction in the width of the affected retina. Considerable retinal and cerebellar development occurs postnatally in the rat, thus newborn animals might be useful in the testing of possible teratogenic drugs.


The zonular insertion: a scanning electron microscopic study. The zonular insertion on the human lens capsule has been studied with scanning electron microscopy. The whole insertional area is covered by a variety of 35 to 55 nm. fibrils, including those of the major inserting zonules, the meridional zonules, capsular surface fibrils, and a nonoriented fibrillar layer over the zonules. All appear to be zonular in type. The meridional zonules span the equator and are a form of insertion shared by the three levels of major zonules, probably helping to distribute their lateral pull. Surface rippling over the zonular insertion appears to result from greater contraction of the oriented fibers than of the nonoriented fibrils and superficial capsule. Surface meshwork fibrils attach to the capsule by becoming embedded in a granular capsular matrix 0.5 to 1 mu thick.


Intracellular crystalline deposits by bacteria grown in urine from a stone former. Several species of bacteria were found to form an intracellular crystalline material when grown in urine obtained from a subject with a history of infrequent renal calculi formation. The following species: Proteus mirabilis, Proteus rettgeri, Providencia stuartii, Enterobacter aerogenes, Enterobacter cloacae, Escherichia coli, and Candida albicans formed crystals of hydroxyapatite. Klebsiella pneumoniae, Pseudomonas aeruginosa, and Proteus vulgaris produced crystals of calcite -II. Several of these bacteria have been isolated from the kidneys of patients with renal caculi indicationg that microorganisms may be involved in the nucleation process during calculogenesis.


The role of trace metals in calcium urolithiasis. Ten urinary stones composed of calcium oxalate or a mixture of calcium oxalate and calcium phosphate were analyzed for trace metal content by emission spectroscopy. Trace metals found in amounts of 0.001 per cent or more were iron, copper, zinic, tin, lead, and aluminum. The inhibitory effect of each of these trace metals on the crystal growth of calcium oxalate and calcium phosphate was tested. Results indicated that none of the metal affect the crystal growth of calcium oxalate at concentrations approximating those found in normal urine. The metal ions copper (II), zinc (II), tin (II), and aluminum (III) did affect the crystal growth of calcium phosphate when present at physiologic concentrations; however, their contribution to the total calcium phosphate inhibitor activity in urine was estimated to be insufficient to have a regulatory role in urinary stone growth.


Effects of androgen and estradiol administration on the weight of the ventral prostate, seminal vesicles, and testes of immature rats. This study was designed to investigate the effects of pharmacologic doses of estradiol on th biologic actions of testosterone and 5alpha-dihydrotestosterone when administered to immature (21 to 28 of age) male rats. The weights of the prostate, seminal vesicles, and testes were used as the indicators of androgen effects. Estradiol had no effects on the testosterone- or 5alpha-dihydrotestosterone-induced increases in prostate weight over the 12 days after the androgen injection. Estradiol acted synergistically with testosterone but not 5alpha-dihydrotestosterone on the increases in seminal vesicle weight. Estradiol had no effect, by itself, on the rate of testes growth. Both androgens resulted in a decreased rate of testicular growth and this was not infuenced by estradiol.


Gross and radiologic anatomy of the canine kidney. Review of medical and veterinary literature reveals no complete gross or radiologic description of the canine kidney. Yet, this organ is often the recipient of urologic investigation. Therefore, a complete gross study of this organ was carried out and the results are presented.


Effects of crush and devascularizing injuries to the proximal ureter. An experimental study. Crush and devascularizing injuries were performed on proximal canine ureters. The pathogenesis of the reaction to these injuries was evaluated by using microbiologic, physiologic, radiologic, biophysical, histologic, and anatomic techniques. The results indicated that within 12 weeks after crush injuries lasting up to 60 min, renal function fully returned and radiologic demonstration of the crush area resolved, even through other radiologic and pathologic changes persisted in hte crushed ureters. The pathophsiologic results of devascularizing injuries that did not resolve spontaneously tended to progress more rapidly and case more serious renal impairment than any of the crush injuries. Devascularizing injuries that resolved tended to be benign in their effect.


Bacillus Calmette-Guérin and dinitrochlorobenzene immunotherapy of chemically induced bladder tumors. Immunologic incompetence is correlated with increased susceptibility to malignacy. Immunostimulation with such agents as Bacillus Calmette-Guérin (BCG) and dinitrochlorobenzene (DNCB) have resulted in regression of cutaneous maligancies. An evaluation of the effect of BCG and dinitrochlorobenzene sensitization and bladder irrigation in Fischer rats revealed no decrease in the incidence of bladder tumors induced by 2-formylamino-4-(5-nitro-2-furyl) thiazole (FANFT). However, direct intralesional injection of BCG into early bladder tumors was effective in preventing the progression of such tumors in seven of ten BCG-sensitized rats (P less than 0.005). No effect was seen when BCG was injected intralesionally in unsensitized rats or when control solutions were injected.


Cryoimmunologic investigations of the rabbit kidney. The unilateral kidneys of rabbits were frozen in situ for 3 min at --160 C with liquid nitrogen. After freezing, proteinuria was observed in a few rabbits. Passive hemagglutination and Ouchterlony double diffucion tests confirmed the presence of circulation antibodies against renal extracts in the serum. Immunofluorescent studies showed deposits of IgG along the renal glomerulus and in the mesangium. However, light microscopy results showed no change in renal glomerulus and electron microscopy results showed no depositions in the glomerular basement membrane. During 5 months after freezing of the kidney, glomerulonephritis has not occurred clinically and pathologically.


Measurement of plasma contrast levels during excretory urography. Ultraviolet spectrophotometry permits easy measurement of plasma levels of intravenously injected contrast media. The initial results are described from the determination of plasma methylglucamine iothalamate (Conray 60) levels during routine excretory urography in 10 normal patients. This simple procedure has many potential applications in the investigation of contrast media physiology.


Experimental basis for intraoperative calculus localization by ultrasound. Experiments were conducted in order to prove the usefulness of ultrasound, B-Scan and A-Mode, and A-Mode alone in the localization of small (3 to 10 mm) fragments of foreign materials in renal substance. Clinical experience indicates that stone fragments as small as 3 mm can be localized intraoperatively using A-mode alone, sterilized with gas for use in a surgical field.


The structure of the rabbit urinary bladder after experimental distension. The effect of prolonged bladder distension produced by elevating intravesial pressures under controlled conditions has been examined in rabits. Bladders were distended in vivo for 3 hr and studied at selected intervals during the postoperative period using light and electron microscopic techniques. In the acute phase, the continuity of the epithelium was partially destroyed and submucosal hemorrhages and edema were evident. However, these changes rapidly resolved and speciments obtained 7 days after distension were similar to controls. Using electron microscopy, smooth muscle cells were structrually unaffected by the experimental procedure and "junction" regions between adjacent cells were of normal appearance throughout the postoperative period. The arrangement and fine structure of autonomic nerves amongst detrusor smooth muscle were similar to those of control samples. These findings are discussed with reference to the findings of other workers and in relation to the use of bladder distension in the treatment of human detrusor instability.


The role of intrarenal angiotensin II in the hemodynamic response to unilateral obstructive uropathy. Intrarenal infusion of Sar1-Ala8 angiotensin II(P113), a competitive antagonist of angiotensin II, failed to reverse or prevent the decreased ipsilateral renal blood flow and increased renal resistance characteristic of chronic unilateral ureteral occlusion and did not significantly alter plasma renin activity in the chronic state. Thus, angiotensin II does not appear to play a significant role in the hemodynamic response to unilateral ureteral occlusion.


Effects of prolactin on the dorsolateral lobe of the rat prostate gland. Intact mature young rats (8 to 10 weeks old) or older rats (16 to 18 weeks old) were used to study the effects of various doses of prolactin on the dorsolateral lobes of the prostate. Prolactin (50 or 150 IU per kg) administered subcutaneously for 5 days in intact mature young rats caused increases in wet weight, protein levels, and nucleic acid levels in the dorsolateral lobes of the prostate. Similar results were obtained when rats were given injections of prolactin for a period of 10 days. When injecting only a 5 IU per kg dose of prolactin, no significant changes were evident in the tissue weight, protein levels, or nucleic acid levels of the prostate gland; however, this same dose produced increases in these biochemical parameters in the dorsolateral lobe of the prostate of older rats.