Comparison of cream of rice agar and horse serum for differentiating germ tubes of Candida albicans from filaments of Candida tropicalis. Simple cream of rice agar was superior to horse serum for the demonstration of germ tubes by Candida albicans and in the differentiation of pseudohyphae of Candida tropicalis from germ tubes at 37 degrees C. Mycelium and chlamydospores were also produced on this medium.


Influence of a deficiency of the second component of complement on the bactericidal activity of neutrophils in vitro. Serum from three patients with a complete, selective deficiency of the second component of complement (C2) did not promote optimal killing of Staphylococcus aureus, 502A by neutrophilic polymorphonuclear leukocytes (PMN) in vitro. The addition of C2 reagent or the presence of heat-stable opsonin in the C2-deficient serum corrected the defective killing of S. aureus that was observed with patient or control PMN. PMN from the patients or control subjects killed bacteria with equal efficiency under conditions of optimal opsonization (normal pooled serum). However, twice-washed control PMN were better than patient PMN in killing S. aureus under circumstances of suboptimal opsonization (C2-deficient serum, heated C2-deficient serum, heated normal pooled serum, or no replacement of serum). The latter finding was due to residual C2 on the surface of twice-washed control cells. As repeated washing of control PMN progressively removed cell-associated C2, the staphylocidal effectiveness of the control RMN decreased to the level of patient PMN. In contrast to the findings with S. aureus, triply-washed PMN from patients or controls killed normal numbers of Escherichia coli, ON2, in C2-deficient serum.


Isolation and functional characterization of human intestinal mucosal lymphoid cells. Viable suspensions of human colonic mucosal lymphoid cells have been prepared by sequential treatment of tissue with dithiothreitol, EDTA in calcium- and magnesium-free salt solutions, and purified collagenase. The intestinal lymphocyte population, in comparison with that of peripheral blood, had greater numbers of bone marrow-derived cells, particularly cells bearing membrane IgA; showed spontaneous association with macrophages; underwent rapid rosette formation with sheep erythrocytes; and demonstrated increased in vitro synthesis of immunoglobulin. Total thymus-derived cells were equal in the two populations. Decreases were found in "null" cell numbers, in cells bearing membrane IgD and IgM, and in responsiveness to phytohemagglutinin. Macrophage/monocytes in the intestinal population were increased in size, granularity, motility, sustained glass adherence, and phagocytic activity. Human intestinal lymphoid cells appear to constitute a cell population that is more "mature" and/or "activated", in comparison with the lymphoid cells of peripheral blood. The method of preparation should lend itself to the study of inflammatory bowel disease, gastrointestinal cancer, and the intestinal secretory immune system.


The body image of pregnant women as reflected in their human figure drawings. Drawings of female figures made by women in their first (N = 54), second (N= 51), and third (N = 56) trimesters of pregnancy and post-partum (N = 55) women were compared with each other and with a control (N = 76) group of gynecological patients. There were no major differences in the drawings of women during the three major stages of pregnancy or between the pregnant women and those who had delivered recently. However, the pregnant women differed significantly from the gynecological controls in that they made more nude drawings, emphasized the genitals, distorted the drawn figures, and made the drawings smaller in size. It was concluded that somatic and psychological changes associated with pregnancy are reflected in the human figure drawings.


Detection of immune complexes. The use of radioimmunoassays with Clq and monoclonal rheumatoid factor. This study describes two sensitive, rapid, relatively simple, competitive inhibition radioimmunoassays for detecting immune complex. The tests are based on the inhibition of I125-Clq or I125-monoclonal rheumatoid factor (mRF) binding to an insoluble substrate, IgG-Sepharose. The assays can be performed in 5 h utilizing 10 micronl of serum. Heating of serum is not required and polyclonal rheumatoid factors do not interefere. With the two assays, a wide range of complexes of various size and complement fixing activity can be detected. The Clq test can detect complement fixing Ig complexes larger than 19S, while the mRF tests detect complexes of IgG as small as 8S irrespective of their complement fixing activity. Mouse, rabbit, and human aggregated IgG (agg IgG) can be detected in the Clq test, and human and rabbit agg IgG in the mRF test. As low as 4 microng/ml of isolated human agg IgG can be detected in the Clq test and 0.5 microng/ml in the rheumatoid factor test. Sensitivity is greater for mouse agg IgG. For pathologic sera which must be diluted to eliminate interfering factors, the sensitivity of the assay is approximately 10 times less. The Clq test showed marked inhibition by systemic lupus erythematosus sera with close correlation with CH50 levels and disease activity. The mRF test showed better correlation with rheumatoid arthritis sera. In addition, anionic macromolecules known to react with Clq and other Clq reactants that occur in pathologic sera such as the "low molecular weight" substances in systemic lupus erythematosus are also detected. These reactants are not detectable in the mRF test and can be eliminated in the Clq test by performing the test at higher ionic strength. The tests can be applied to the study of a variety of pathologic states where immune complexes appear to play a role.


Characterisation of immunofluorescent heterophile antibodies which may be confused with autoantibodies. Approximately 6% of sera submitted for routine immunofluorescent autoantibody screeening produced characteristic reaction patterns against a variety of animal and human tissues. It is suggested that these non-tissue-specific patterns represent a complex family of heterophile antibodies which could be confused with certain autoantibodies. It is further suggested that these heterophile antibodies bear a relationship to IgG isohaemagglutinins.


Pathology of 'non-healing (midline) granuloma'. In a histological study of biopsy and postmortem material from 30 cases of nasal disease in which a clinical diagnosis of 'midline granuloma' or Wegener's granuloma had been given, we selected 10 cases on the basis of the presence of widespread coagulative necrosis and atypical cells. Evidence is provided that such changes represent a malignant neoplasm of histiocytic lymphoma type. Local invasion and spread to cervical and more distant lymph nodes, spleen, liver, and kidney were seen in some of the cases. Erythrophagocytic activity was marked in the spleen in three cases and histiocytic infiltration of the bone marrow in two cases, indicating a more generalised activity of histiocytic cells. Terms such as 'malignant granuloma' should be abandoned. In obstructive and ulcerating conditions of the nose efforts should be concentrated on making an accurate histological diagnosis.


Test reproducibility of the API (20E), Enterotube, and Pathotec systems. Thirty-three strains of bacteria (30 Enterobacteriaceae and one strain each of Aeromonas formicans, A. hydrophila, and Plesiomonas shigelloides) were tested three times in each of 27 conventional tests and in the API, Enterotube, and Pathotec systems. The results obtained were analysed for test reproducibility within each kit, correlation of the kit tests with the equivalent conventional media, and the identification of the strains by the kits. Difficulties in evaluation and comparison of identifications are discussed. A practical evaluation of the kits was also made.


The clinical pharmacology of methylprednisolone sodium phosphate. I. Intramuscular route of administration. Intramuscularly administered methylprednisolone sodium phosphate (Medrol Stabisol) in single doses of 40, 80, or 160 mg (methylprednisolone equivalents) had a similar effect as the same doses of methylprednisolone sodium succinate (Solu-Medrol) with regard to eosinophil suppression, elevation of glucose, white blood count differential shifts (lympholytic effect), urinary excretion of sodium and potassium, and localized (pain) and systemic side effects. The average plasma methylprednisolone concentration was approximately 20% higher after the intramuscular administration of methylprednisolone sodium phosphate than after methylprednisolone sodium succinate. The differences in plasma methylprednisolone levels produced by the two esters suggest that either hydrolysis of the succinate ester occurs more slowly or the succinate ester distributes more extensively. This difference in plasma level, however, is not reflected in any other pharmacologic evaluation of the two esters, e.g., both eosinophil depression and hyperglycemic response were identical. No clinically significant changes in the vital signs, standard hematology, and clinical chemistry parameters evaluated were noted after 21 successive doses (q.i.d. for five days with one dose in the morning of day 6) of 80 mg methylprednisolone sodium phosphate. An increase was noted in the systolic blood pressure from a pretreatment mean of 113 mm Hg to a posttreatment mean of 123 mm Hg and an increase in the body weight from a pretreatment mean of 177 pounds to a posttreatment mean of 183 pounds. No signs of adrenal suppression were found as judged by plasma cortisol and ACTH levels. Six (6/12) subjects of the methylprednisolone sodium phosphate group, one (1/12) subject of the vehicle group, and one (1/12) subject of the placebo (sterile saline) group reported the following systemic side effects: gas in stomach, headaches, anorectal itching, and dryness of itching of the skin. No trend was observed for any side effect reported. In these double-blind, randomized studies, single (40, 80, and 160 mg) and multiple (80 mg) intramuscular doses of methylprednisolone sodium phosphate were tolerated in healthy volunteers as well as the same doses of methylprednisolone sodium succinate and similar volumes of vehicle or placebo.


Double-blind comparison of triamterene plus hydrochlorothiazide and spironolactone plus hydrochlorothiazide in the treatment of hypertension. The results from this double-blind, multi-investigator study showed that a combination of 50 mg triamterene plus 25 mg hydrochlorothiazide and a combination of 25 mg spironolactone plus 25 mg hydrochlorothiazide were equally efficacious in lowering blood pressure in hypertensive outpatients, and that they produced the same type and incidence of adverse effects. Likewise, the two drug combinations produced similar effects on blood chemistry and hematology. There were no significant differences between the two combination drugs in efficacy laboratory studies, or adverse effects.


Influence of age, dietary protein and weaning on calf abomasal enzymic secretion. Using fistulated calves, the influence of the age, type of dietary protein and weaning on the secretion of chymosin and pepsin by the abomasum were studied. The abomasum secreted both chymosin and pepsin when the animals were fed milk. Chymosin secretion appeared to be independent of the age of the animals whereas a slow increase in pepsin secretion was observed as the calves aged. Several preruminant animals were fed either a skim-milk diet or a milk substitute in which proteins were provided by fish, soya or whey concentrates. Each of these 3 milk substitutes led to a decrease in chymosin secretion without modification of pepsin secretion. Chymosin secretion was partly restored when the claves were again given a skim-milk diet. At weaning, chymosin secretion dropped abruptly, but the pepsin level was not affected. These results indicate that milk (most probably its casein fraction) is responsible for the activation of chymosin secretion.


Antidiarrheal effects of dihydroergotamine. Dihydroergotamine (DHE), an alpha-adrenergic blocking agent, rapidly improved 121 out of 123 diarrheal patients. A hypotonic sigmoid and a hyperreactive rectum were found in these patients. Manometric studies of the distal colon showed that DHE counteracts the rectal hyperactivity and increases sigmoidal tone. On the other hand, anticholinergic drugs and/or emotional stimuli accentuate the rectal hyperactivity of diarrheal patients. Both features could be due to an unbalanced neurologic control of the gastrointestinal tract with dominance of the alpha-adrenergic over the cholinergic activity. Diphenoxylate (DPO) suppressed the diarrhea in two patients not improved by DHE. Furthermore, DPO reinforced the therapeutic success of DHE in 11 lactose intolerance diarrheal patients, suggesting that the two drugs exert their effects by means of different mechanisms.


Effect of a new beta-adrenergic blocker, l-bunolol, on blood pressure and on the renin-aldosterone system. The antihypertensive effect of a new beta-adrenergic receptor blocker, l-bunolol, was evaluated in 11 hospitalized hypertensive patients of whom four belonged to the high-renin, five to the normal-renin, and two to the low-renin subgroup. There was a significant decrease in blood pressure in most patients, often to normal. Moreover, plasma renin levels were nearly always markedly suppressed, and this suppression preceded the fall in blood pressure. While most high and normal renin patients responded, the numbers of patients within each subgroup were too small to correlate the blood pressure responses with changes in renin levels. There was a significant induced decrease in aldosterone excretion, which usually paralleled the renin suppression. Pulse rate was also consistently reduced during treatment. No weight gain was observed, except in one patient who developed overt congestive cardiac failure. No other side effects were recorded. Ii is concluded that l-bunolol is another beta-blocking drug with significant antihypertensive action. It was found to be effective in cases that were resistant to propranolo. It is well tolerated and safe to use, provided that early evidence of fluid retention is sought for and treated immediately.


An improved technique for the retention of polycarbonate crowns. The following conclusions may be drawn from this study: Composite resins do not bond to polycarbonate crowns. A technique for priming the internal surfaces of polycarbonate crowns is indicated, in order to create such a bond. Priming the polycarbonate crown with methyl methacrylate monomer alone produces a bond with composite resins. Priming the polycarbonate crown with a syrup of methyl methacrylate monomer and (poly) methyl methacrylate powder appears to produce a stronger bond with composite resins than the monomer alone.


Prosthetic management of palatopharyngeal incompetency for the pediatric patient. The diagnosis and treatment planning for palatopharyngeal incompetency is such that decisions can best be arrived at following a qualified interdisciplinary team assessment. The pedodontist, as an integral member of a rehabilitation team, should be knowledgeable in the area of speech prostheses for the child patient. It is only through an increased knowledge and understanding of current surgical and prosthetic techniques that patients with palatopharyngeal incompetency can receive better treatment.


The effect of fluoride impregnated dental floss on enamel fluoride uptake in vitro and streptococcus mutans colonization in vivo. The conclusions reached from this investigation can be summarized as follows: Fluoride can be incorporated into unwaxed dental floss. Placement of the fluoride impregnated dental floss in acid-buffer solution results in the release of most of the fluoride in the floss. Interproximal surfaces of teeth treated in vitro with fluoride impregnated dental floss acquired significantly (approximately three fold) more enamel fluoride than those treated with plain dental floss. The number of in vivo interproximal areas harboring Streptococcus mutans was reduced significantly after treatment with fluoride impregnated dental floss. Further studies should be done to establish the biological, physiochemical, manufacturing, and practical aspects of fluoride impregnated dental floss.


Reduction of dental decay in rampant caries individuals following short-term kanamycin treatment. A week of kanamycin gel treatment before and after the placement of dental restorations, compared to a placebo gel treatment, significantly reduced the levels of cultivable bacteria, S mutans and S sanguis, in the plaque samples collected immediately after the completion of the gel treatments, and was associated with a 46% reduction in new carious surfaces in the 14- to 37-month period following the gel treatment.


Detection of functional complement components in gingival crevicular fluid from humans with periodontal diseases. Crevicular fluid was collected from patients with periodontitis by a capillary tube procedure. Complement component activities were determined by functional assay systems, with human complement, and partially purified human first complement component (C1) as  controls. The complement-fixing properties of the dental plaque of each patient were also examined C1 activity in the crevicular fluid of all patients was approximately 1/8 of whole serum C1 and diminished rapidly with time after collection. There was no significant relationship between C1 concentration and crevicular fluid flow rate. Hemolytic activity of whole complement was also invariable detected when sufficient amounts (8 micronl) of crevicular fluid could be obtained. Dental plaque was found to fix C1. A role for crevicular complement in inflammatory periodontal disease is suggested.


Investigation of adhesive properties of dental composite materials using an improved tensile test procedure and scanning electron microscopy. A standardized tension test was used to evaluate the adhesive properties of several composite materials when used on both dentin and enamel specimens. The nature of the test surfaces was examined by roughness tests using a Talysurf machine and also in more detail by means of a scanning electron microscope. Poor results were obtained for the adhesion of composite materials to dentin whereas good retention to enamel was obtained.


Otologic complications following the use of a high-speed air-turbine handpiece. The use of high-speed instruments may result in subcutaneous air in the face and neck. Hearing loss and otalgia have been added to the previously reported sequelae. Careful use of air-driven high-speed handpieces around areas of weakened and disrupted oral tissues may help avoid subcutaneous emphysema. Awareness of this entity plays a significant role in appropriate diagnosis and treatment.


Use of teeth with a poor prognosis in cleft palate prosthodontics. Patients who have unoperated or incomplete closure of their clefts and who have only a few teeth of poor prognosis remaining are a challenge to the prosthodontist. Two cases have been presented to illustrate treatment techniques that can be used for such patients.


Effect of bronkosol and its components on cardiopulmonary parameters in asthmatic patients. Bronkosol and its components, isoetharine hydrochloride and phenylephrine hydrochloride, were compared in a double-blind randomized fashion for their effect on pulmonary and cardiovascular parameters in patients with reversible bronchospasm. Bronkosol and isoetharine produced significant bronchodilatation as measured by forced expiratory volume in one second (FEV), forced vital capacity (FVC), and mean forced expiratory flow during the middle half of the FVC (isovolume FEF 25% to 75%), and phenylephrine did not. There was no difference between Bronkosol and isoetharine in the degree or duration of bronchodilatation. Significantly more patients had to "discontinue" after 15 min on the day of testing with phenylephrine than with Bronkosol or isoetharine. Heart rate was not increased after Bronkosol or isoetharine, confirming its selective beta-2 action. The addition of phenylephrine to isoetharine had no beneficial effect on oxygen saturation. There was no significant difference between these drugs in systolic and diastolic blood pressure, pulse, or respiration. The results of this study cast doubt on the useful contribution of phenylephrine in Bronkosol.


Science--neglected ingredient of nutrition policy. 10th Martha F. Trulson Memorial Lecture. Programs to alleviate malnutrition in children in developing countries need revision. Intervention field trials in Thailand, Tunisia, and Guatemala, based on amino acid fortification and supplementary vitamins and minerals, have had little effect on children. In fact, it is often a misconception that frank deficiencies are common characteristics of malnutrition in developing countries.Rather, stunted growth--caused by caloric deficiency often in the presence of adequate food supplies--may be the most prevalent form of malnutrition. The situation occurs when the customary staple food--for instance, rice in Thailand--has such a high caloric density that children cannot eat enough food to meet their needs. Knowledge is not vet available on ways to solve this dilemma. Nevertheless nutritionists must come forward and be willing to contribute their knowledge and expertise in the shaping of national and international nutrition policies to improve the healthand well-being of populations.


Surgical treatment of congenital zygodactylism and brachymetapody: a case history. This paper has presented a case history of congenital zygodactylous feet with unilateral brachymetapody. A detailed description was presented of the surgical attempt to treat these anomalies and the postoperative complications that were encountered. Follow-up on this patient after 1 1/2 years have shown an excellent result.


Colloidal gold, a useful marker for transmission and scanning electron microscopy. Electron dense markers of a size suitable for transmission electron microscopy and scanning electron microscopy have been prepared with gold granules labeled with a monolayer of specific macromolecules. The optimum conditions for preparing the markers have been ascertained. The method is simple, rapid and seems to be general since gold granules have been labeled with polysaccharides and proteins. As homogeneous populations of gold granules having different sizes can be prepared, the method is also suitable for double marking experiments. The gold technique is illustrated by the localization of polysaccharides and glycoproteins on yeast cell walls and erythrocyte membranes by transmission electron microscopy and on yeast cells and intact erythrocytes by scanning electron microscopy. Good spatial resolution of the marker was achieved in all cases. The method is also suitable for marking thin sections. Spectrophotometric measurements were used to determine the number of gold granules adsorbed per cell.


Functional heterogeneity in macrophages activated by Corynebacterium parvum: characterization of subpopulations with different activities in promoting immune responses and suppressing tumor cell growth. Peritoneal cells (PEC) from mice injected i.p. with heat-killed Corynebacterium parvum (CP) showed enhanced immunostimulatory (accessor or A cell) activity as measured by their ability to restore the immune responsiveness of nonadherent spleen cells to sheep erythrocytes (SRBC) and polymeric flagellin (POL) of Salmonella adelaide in vitro. This was true whether the PEC and nonadherent spleen cells were in direct contact or separated by a cell-impermeable membrane which allowed the free passage of soluble mediators. CP-activated PEC also exhibited greatly increased cytostatic activity against the growth of syngeneic tumor cells in vitro. After fractionation of the PEC according to cell size by velocity sedimentation, a separation of A cell activity from anti-tumor activity was observed. Although both these functions were associated with phagocytic cells of the monocyte-macrophage series, the highest A cell activity was found in fractions containing small and medium-sized macrophages, whereas the anti-tumor activity increased with cell size to a maximum with the largest macrophages. Thus, there is a relative increase of suppressive activity over stimulatory activity with an increase in cell size. Cytochemical and morphologic evidence suggests that the A cell-rich fractions contained small and medium-sized macrophages which were derived from newly arrived monocytes, whereas the large tumor-suppressive macrophages were relatively more differentiated.


Characterization of the nonphagocytic adherent cell from the peritoneal cavity of normal and BCG-treated mice. A distinctive subpopulation of nonphagocytic, tightly adherent cells (NPAC) comprised approximately 6% of the adherent peritoneal cells from untreated mice, and about 18% of those from mice previously given BCG i.p. A separation procedure based on adherence and lack of phagocytosis was devised. Isolated NPAC were morphologically intermediate between small lymphocytes and macrophages. They were positive for nonspecific esterase, negative for peroxidase, positive for surface IgM, and negative for surface IgG1, IgG2 and IgA. When capped, their surface IgM regenerated in vitro. NPAC had demonstrable Fc receptors but not EAC receptors. They resisted killing by an anti-macrophage serum, were negative by immunofluorescence with an anti-T cell reagent, and incorporated increased amounts of thymidine in response to LPS but not to PHA. They were more readily killed with anti-Ia serum and complement than macrophages, but less readily than splenic B cells. NPAC appeared to represent a subpopulation of B lymphocytes which contaminates some preparations previously regarded as "macrophages" and which may be ressponsible for some of the activities previously ascribed to "macrophages".


Antigen modulation of antibody forming cells: the relationship between direct plaque size, antibody secretion rate and antibody affinity. Using the mathematical theory of direct plaque growth, we have analyzed the expected variation of plaque size with IgM affinity and secretion rate. We use the theory to comment on recent effector cell blockage experiments and show how the theory can be used to determine the change in the secretion rate of a single antibody-forming cell subjected to blockage by a multivalent antigen. We also argue, using the mathematical theory, that under the usual experimental conditions employed in the plaque assay, cells that produce low affinity IgM antibodies will give rise to smaller plaques than cells that produce high affinity IgM antibodies.


In vitro augmentation of lymphocyte sheep cell rosette formation by leukocyte dialysates. We have studied in vitro the effects of leukocyte dialysates containing transfer factor on the formation of sheep cell rosettes by human lymphocytes. Dialysate had no effect on the toral rosettes, but increased the number of rapidly forming ("active") rosettes. This was due to an increased affinity of the lymphocytes for the sheep red cell. Trypsin-treated lymphocytes regained the ability to form rosettes more rapidly when cultured with leukocyte dialysates than with control media. These experiments suggest that leukocyte dialysate acts to increase the number or arrangement of sheep cell receptors on the lymphocyte surface.


Expression of alloantigens LY-5 and LY-6 on cytotoxic effector cells. With anti-Ly antisera and complement it has been possible to demonstrate that cytotoxic effector cells generated in vitro against allogeneic cells carry the alloantigen Ly-5 and Ly-6. The studies show that antisera directed against the Ly-6 antigens, together with complement, eliminate essentially all of the killer cells, of the appropriate strain, while killing only 50 to 60% of Thy-1+ cells. Anti-Ly-5 antsera and complement lysed 55 to 65% of Thy-1+ cells and killed a significant portion, but not all, of the cytotoxic cells. The later finding was investigated in more detail since it suggested a degree of heterogeneity within the killer cell subpopulation. However, the data did not support that conclusion, at least for allogeneic killer cells.


Complement-dependent and -independent pathways of T cell-B cell cooperation. BDF1 mice treated with CoV had markedly reduced levels (less than 20%) of native serum C3 32 hr later, whereas the frequency of splenic CR+ cells was normal. CoV treatment before immunization reduced the IgM PFC response to a T-dependent antigen (TNP-SRBC) by more than 60%. Inclusion of highly specific anti-C3 antibody had no effect on the T-dependent IgM response of CR- B cells. The residual PFC responses in cultures of unfractionated spleen cells treated with anti-C3 could be largely or completely accounted for by CR- B cells in the cultures. The effect of anti-C3 antibody was not due to cytotoxicity. These data collectively indicate that the effect of CoV on T-dependent antibody responses is due to decreased C3 in serum rather than to interaction of C receptors directly with CoV or with C3 cleavage products. They suggest the existence of at least two distinct pathways of T-B cooperation, one in which C3 is an obligatory participant and another in which it may be uninvolved.


Evidence for a B lymphocyte defect underlying the anti-X anti-erythrocyte autoantibody response of NZB mice. The autoimmune hemolytic anemia of NZB mice is pathogenetically mediated by a genetically prescribed anti-erythrocyte autoantibody response directed to the X erythrocyte autoantigen. The cellular locus of the immunoregulatory defect underlying the anti-X response was explored by adoptively transferring bone marrow cells (BMC) from NZB mice to lethally irradiated histocompatible recipients. Before adoptive transfer, BMC from donor mice were assayed for antigen-binding lymphocytes with receptors for the X autoantigen (X-ABL) by immunocytoadherence assays and for anti-X autoantibody-secreting cells (X-PFC) by plaque-forming cell assays. Twelve weeks after adoptive transfer, splenic lymphocytes from recipient mice were assayed for X-PFC and humoral anti-X autoantibody by Coombs' tests. Transfer of 15 to 30 x 10(6) BMC containing 6 to 12 x 10(3) X-ABL but no X-PFC from 6- to 8-week-old NZB mice to lethally irradiated BALB/c, B10.D2, C57BL/Ks, and DBA/2 mice produced X-PFC in 70% of the recipients. Development of X-PFC was not simply dependent upon available X-ABL since transfer of 15-30 x 10(6) BMC, containing comparable numbers of X-ABL, from BALB/c, B10.D2, C57BL/Ks, or DBA/2 mice to NZB or syngeneic recipients did not produce X-PFC. Transfer of BMC from NZB mice to BALB/c, B10.D2, and DBA/2 mice with weekly administrations of AKR anti-theta antiserum had no effect on the development of X-PFC; Tlymphocyte ablation was evidenced by the absence of theta+ spleen cells. These results suggest that the pathogenetic anti-X response is not genetically prescribed at the level of macrophages, humoral factors, or T cells, but rather appears to be a phenotypic expression of a primary B lymphocyte defect permitting or promoting differentiation of NZB X-ABL.


Preferential assignment of allotype a1 globulin for the production of early IgM anti-para-azobenzenearsonate antibodies in heterozygous a1a3 rabbits. Rabbits of allotype a1a3 were injected on days 0, 2, and 4 with mixtures containing equal amounts of pigeon erythrocytes (Prbc) coupled to para-azobenzenearsonate (AA) and to para-azobenzene-N-trimethylammonium (TMA). On day 6, the allotypes of antibody from plaque-forming cells (PFC) of the blood were determined by observing the inhibition of plaque formation by anti-allotype sera. Anti-AA PFC appeared to consist for the most part of cells making antibody of allotype a1 since 65% of them were inhibited by anti-a1 serum and only 8% by anti-a3. Anti-TMA PFC, on the other hand, appeared to consist mostly of cells making antibody of allotype a3, since less than 1% of them were inhibited by anti-a1 but 47% by anti-a3. Antibody allotype for spleen PFC was also determined on day 6 and was similar to that found for blood PFC. Anti-AA PFC were inhibited 74% by anti-a1 serum and 15% by anti-a3 whereas anti-TMA PFC were inhibited 19% by anti-a1 and 43% by anti-a3. Serum hemolysin specific for AA hapten from a1a3 animals was also strongly inhibited by anti-a1 serum but not by anti-a3 whereas the converse was true for hemolysin against TMA hapten. The a1a3 rabbits, in whcih the anti-AA was restricted to allotype a1, were mated to produced homozygous a3a3 animals. When the PFC and serum antibodies of these a3a3 offspring were examined by specific inhibition, the anti-AA activity was found to be of allotype a3 rather than being a-negative. The number of anti-AA PFC in the blood of a3a3 rabbits was lower than that in blood of a1a3 or a1a1 animals. In addition, the TMA hapten appeared to inhibit the response to the AA hapten. Thus a1a3 rabbits immunized with AA-Prbc alone had 14-fold more anti-AA PFC or 18-fold higher anti-AA hemolysin titer than a3a3 animals immunized with both AA-Prbc and TMA-Prbc. Our results are discussed in relation to various explanations which have been offered for an imbalance of allotypes in a given antibody.


A simple fluorescent method to determine complement-mediated liposome immune lysis. A simple inexpective method is described to study the kinetics of complement-mediated immune lysis of liposomes containing sheep red blood cell lipid antigens. It is based on the fact that trapping the fluorescent molecule 1-aminonaphthalene-3,6,8-trisulfonate and the dynamic quencher, alpha, alpha'-dipyridinium p-xylene dibromide within the liposome inner volume results in an extinguished fluorescence signal. On addition of helmolysin plus active complement, liposome lysis occurs. The exit of the fluorophore and quencher and their subsequent dilution in the external volume abolishes the quenching, resulting in a high fluorescence signal. The details of the method are described as well as the initial kinetic results.


Labelling of lymphocytes with various radiosotopes for in vivo tracer studies; a review. Several methods are available for labelling lymphoid cells with radioactive markers e.g. 3H, 14C, 125I, 131I, 35S, 75Se, based on general metabolic processes, involving DNA, RNA and protein synthesis in these cells. Together with methods for labelling lymphoid cells with 51Cr and 99mTc, radioisotopes which are captured in cells but not incorporated in its metabolites, these will be reviewed in the present paper. Attention will be given to possible applications of the use of radiosotope labels.


Use of antibody-coated red cells for the sensitive detection of antigen and in rosette tests for cells bearing surface immunoglobulins. Conditions for an improved chromic chloride method for the attachment of antibody to red cells are described. The method, which is applicable to whole Ig fractions as well as affinity-purified antibodies, is reproducible and has a sensitivity for detection of antigen in the nanogram range. The coated cells may be used in a rosette assay for the detection of cell surface-bound antigens.


Absorption analysis of H-2D and K antigens on spermatozoa. The presence of H-2D and K antigens on mouse spermatozoa has been investigated by absorption followed by testing on the proper target lymphocytes. It is concluded that, in addition to Ia antigens, H-2D and K antigens are indeed expressed on mouse sperm cells.


The involvement of a suppressor mechanism in neonatally induced allograft tolerance in mice. The possible involvement of a suppressor cell mechanism in neonatally induced allograft tolerance was tested in two different mouse-strain combinations. In an H-2 incompatible donor-recipient combination presented by the congeneic strains B10.D2 and B10.D2(M504), adoptive transfer of tolerance with 5 X 10(7) spleen cells to syngeneic sublethally (400 R) irradiated recipients was successful resulting in a markedly prolonged test graft survival or even take. The result of tests for negative tolerance (which should be suceptible to abolition by means of cells from normally reactive donors) did not indicate that neonatally induced tolerance across the given barrier rested on this principle. The suppressor mechanism seemed to be involved in neonatally induced allograft tolerance also in the other strain combination (B10-40NX).


Unexpectedly high frequency of antibody to Mycoplasma pneumoniae in human sera as measured by sensitive techniques. Mycoplasmacidal and radioimmunoprecipitating antibodies to Mycoplasma pneumoniae were commonly detected in healthy children younger than five years of age, in whom natural disease due to M. pneumoniae is rare. Antibody in serum could also be demonstrated in Marine recruits several weeks or months before development of pneumonia due to M. pneumoniae. The latter observation suggests that serum antibody does not confer protection against disease caused by M. pneumoniae.


Colonization of porcine small intestine by Escherichia coli: colonization and adhesion factors of pig enteropathogens that lack K88. The colonizing and adhesive attributes of enterotoxigenic acapsular and/or nonpiliated mutants from K88-negative enteropathogenic Escherichia coli strains were compared with their capsulated and piliated parents (parents were piliated when grown in vitro and in vivo). Acapsular, nonpiliated mutants from three different colonizing strains of enteropathogenic E. coli lost their ability to colonize the ileum of newborn pigs. Acapsular, piliated and capsular, nonpiliated mutants were derived from one of the parental strains (987), and both mutants lacked the ability to colonize the ileum of pigs. The only mutants available from a fourth strain (431) were acapsular and piliated, and they colonized as well as their parents. These data indicate that both capsule and pili are involved in colonization by strain 987. In contrast, capsule is not required for colonization by strain 431, but pili may be.


The pathogenesis of Shigella diarrhea. VI. Toxin and antitoxin in Shigella flexneri and Shigella sonnei infections in humans. Two strains of Shigella flexneri and one of Shigella sonnei were studied for toxin production in vitro. All of the three strains produced a cell-free cytotoxin that showed marked similarity to that produced by Shigella dysenteriae 1. Each toxin eluted in two distinct peaks on chromatography with Sephadex G-150, was destroyed by heating at 90 C for 30 min, and was neutralized by S. dysenteriae 1 antitoxin. Patients with infections due to S. flexneri and S. sonnei developed antibody that neutralized S. dysenteriae 1 toxin in vitro. In three of seven positive sera studied by sucrose density-gradient ultracentrifugation, antibody activity was associated only with the IgM fraction. The time course of the antibody response resembled that found in infections due to S. dysenteriae 1, in which an IgM antitoxin antibody has also been described. Since three species of Shigella have now been shown to be toxigenic, it is possible that bacterial toxin may play a role, along with bacterial invasion, in the pathogenesis of infections due to S. flexneri and S. sonnei, as well as those due to S. dysenteriae 1.


Significance of antibody-coated bacteria in urinary sediment in experimental pyelonephritis. The sequential appearance of antibody-coated bacteria in urinary sediment was followed in experimental pyelonephritis produced with Escherichia coli O6:K13:H1. It was possible to determine against which antigen of the organism this antibody was directed. Antibody-coated bacteria appeared by day 11 of infection, a time which coincided with the synthesis of local (intrarenal) antibody. Bacteria coated with antibody more than three days after serum antibody but four days before urinary antibody appeared. Antibody eluted from coated bacteria was directed against the O-antigen of the infecting organism but not the K-antigen. Newly synthesized intrarenal antibody and urinary antibody were directed only against O-antigen, but not against K-antigen; this finding would explain why the antibody that coated the bacteria was antibody to lipopolysaccharide. Thus, a positive test for antibody-coated bacteria indicates that a local immune response to O-antigen has occurred. It is postulated that this immune response could relate to the clinical symptomatology of acute symptomatic pyelonephritis since most patients presenting with acute pyelonephritis have bacteria coated with antibody.


Characterization of the Chinese hamster ovary cell assay for the enterotoxins of Vibrio cholerae and Escherichia coli and for specific antisera, and toxoid. The morphologic change in Chinese hamster ovary (CHO) cells that is associated with increased intracellular concentrations of adenosine 3':5'-cyclic phosphate (cyclic AMP) provided a highly sensitive assay for the heat-liable enterotoxins of Vibrio cholerae and Escherichia coli. The similarity between the responses of CHO cells and those of intestinal mucosa to enterotoxins was demonstrated by the inhibition of these effects by the ganglioside galactosyl-N-acetylgalactosaminyl [sialosyl] lactosyl ceramide (GM1), as well as by specific antisera. The antisera against either enterotoxin were more effective in inhibiting the effects of homologous toxin than those of heterologous toxin, but cross-neutralization also occurred in both cases. When choleragenoid, the natural toxoid of cholera toxin, was incubated with the CHO cells, the subsequent effects of the toxin on the morphology of the cells were prevented as were the effects of the toxin on intestinal secretion. However, choleragenoid did not prevent the effects of E. coli toxin. The similarity between the action of enterotoxins on CHO cell morphology and on secretion by the small intestine suggests that there are similar binding sites in these tissues. The potential usefulness of the CHO cell assay in quantitative measurement of antitoxic immunity is thus demonstrated.


Clinical and laboratory assessment of the pathogenicity of serotyped enteropathogenic Escherichia coli. Only one of 167 separate isolates of enteropathogenic Escherichia coli (EEC) was shown to produce enterotoxin, and none of the 167 isolates were invasive. Clinical features of 123 hospitalized children with EEC were compared with those of 917 infants with nonbacterial gastroenteritis and 145 infants infected with Shigella. The average duration of diarrhea (five or more stools per day) in hospitalized children with EEC, nonbacterial gastroenteritis, Shigella flexneri, and Shigella sonnei was 4.6, 2.4, 5.1, and 2.5 days, respectively. The average duration of fever in these four groups was 1.4, 1.2, 2.1, and 1.2 days, respectively. The difference in duration of diarrhea between children with EEC and those with nonbacterial gastroenteritis was significant (P less than 0.001), even when age and rural/urban origin were controlled by analysis of variance. Nevertheless, the EEC group tended to be younger and to have a higher proportion of infants of rural origin. Although it appears that EEC serotypes rarely identify invasive or enterotoxin-producing organisms, clinical features of infants with EEC-associated gastroenteritis suggest that these infants may represent a distinctive and clinically important group with gastroenteritis of greater severity than nonbacterial gastroenteritis.


Emergence of resistance to chloramphenicol in salmonella. Surveillance of antibiotic resistance of salmonella isolates from humans in the north-eastern United States has detected resistance to chloramphenicol on eight occasions, three of which were hospital outbreaks. Resistance was developed during treatment in five instances, and in another resistance this situation was suspected. On two other occasions chloramphenicol-resistant Salmonella appear to have been imported from outside the country. Common features of the cases were multiple antibiotic resistance, transfer of R-factor, simultaneous presence of different patterns of resistance, similar patterns of antibiotic resistance in other intestinal bacteria, and apparent in vivo and in vitro loss of resistance to chloramphenicol with time.


Protective effects of passively transferred immune T- or B-lymphocytes in mice infected with Salmonella typhimurium. Purified populations of bone marrow-derived (B-) lymphocytes and thymus-derived (T-) lymphocytes were obtained from C3D2F1 hybrid mice shown to be immune to Salmonella typhimurium. These subpopulations of lymphocytes were injected into normal mice; four days later the animals were challenged with 50 50% lethal doses of S. typhimurium, and viable bacteria in livers, spleens, and blood were counted at various intervals after challenge. On day 8 after challenge, the mice supplemented with B-lymphocytes showed a significant decrease in the number of organisms recovered from all three sites, compared with that seen in recipients of T-lymphocytes and in controls. The mice given B-lymphocytes showed a better rate of survival (65%) than mice that received only T-lymphocytes (21%) or T-lymphocyte fractions contaminated 10%-30% with B-lymphocytes (49%). These data indicate that, although the humoral response is not totally protective, it does play an important role in the suppression of the infection during its early stages.


Quantitative venographic assessment of deep vein thrombosis in the evaluation of streptokinase and heparin therapy. A technique of quantitative venography has been developed in which values are assigned to the deep veins of the calf, knee, thigh, and pelvis, based upon the calculated volume and degree of occlusion of these venous segments. A maximum score of 40 units reflects complete thrombosis of all segments. This technique has been applied to a randomized, single-blind study of streptokinase versus heparin treatment. Each group of 12 patients had similar mean inital venographic scores; follow-up venograms were performed 5 days after the start of therapy. Streptokinase patients with high initial scores (larger than 20) showed a mean improvement of 12.1 units, while those with low initial scores(less than 20) were essentially unchanged. Heparin patients with high scores had a minimal mean improvement of 1.1 units, but those with low scores had a significant mean extension of thrombosis of 8.6 units. Patients with symptoms for less than 7 days showed greated mean improvement (12.7 units) with streptokinase that those with a longer duration of symptoms (2.0 units); heparin patients in these subgroups showed a mean worsening of 7.5 units and no change, respectively. Extrinsic venous obstruction by tumor did not prevent an excellent response to streptokinase. No single test of coagulation of fibriolysis was a reliable indicator of the degree of venographic response to lytic therapy. Pyrexia and hemorrhagic complications occurred in over one-half of the streptokinase patients; one had an anaphylactic reaction, and one died of intracerebral hemorrhage during therapy. The data suggest that lytic therapy is best restricted to the patient with acute extensive thrombosis. Also, continuous infusions of heparin according to current guidelines may be inadequate to prevent thrombus growth in some patients.


Effects of anterior hypothalamic implantation of gonadal steroids on serum and pituitary gonadotrophins (follicle-stimulating hormone and luteinizing hormone) in the castrated male rat. Castrated rats were stereotaxically implanted with 1 microng oestradiol benzoate, 5 microng testosterone isobutyrate or, as a control, 10 microng cholesterol, in the hypothalamus. The effects of the steroids on plasma and pituitary gonadotrophins (FSH and LH) were assessed by radioimmunoassay. Our results indicate that, in the male rat, in addition to the arcuate nucleus-median eminence complex, the preoptic suprachiasmatic area is able to control synthesis and secretion of both gonadotrophins, and that it is sensitive to oestradiol and testosterone.


Steroids and pituitary responsiveness in female, androgenized female and male rats. Spontaneous gonadotrophin release and the gonadotrophin response to LH releasing factor (RF) were studied in pro-oestrus, androgenized female and male rats. The animals were either intact or gonadectomized (about 32 h previously) and treated with various steroids. The gonadotrophin response (especially LH) was much lower in intact males and androgenized females than in pro-oestrous females. Oestrogen plus progesterone increased plasma gonadotrophin concentrations and responses in ovariectomized rats, but inhibited the increase in the plasma gonadotrophin concentration and the LH response which followed castration in males. As in the normal female, ovariectomy decreased the LH response but increased the plasma FSH concentration and response in the androgenized female; oestrogen and progesterone had relatively little effect. Apart from reducing the postcastration rise in plasma FSH, testosterone had no significant effect in gonadectomized male or female animals. These results show that the effect of steroids on the gonadotrophin response to LH-RF as well as the spontaneous secretion of gonadotrophin depends upon sexual differentiation of the hypothalamo-hypophysial system. Studies with various metabolites of progesterone indicated that the facilitatory action of this steroid could be due, in part, to a 5alpha-reduced derivative.


Site of modulatory action of oestrogen and progesterone on gonadotrophin response to luteinizing hormone releasing factor. The effect of oestrogen and progesterone on the spontaneous release of gonadotrophins and on the gonadotrophin response to luteinizing hormone releasing factor has been examined in female rats subjected to hypothalamic deafferentation or pituitary stalk section. The results indicate that both steroids act at the level of the medial basal hypothalamus as well as the anterior pituitary gland; progesterone, for its full effect, also requires the integrity of connexions of the hypothalamus with other areas of the brain.


Hormonal effects of tamoxifen in oligospermic men. The hormonal effects of tamoxifen (10 mg daily for 6 months) have been studied in nine men with oligospermia. Basal concentrations of serum LH (1-7 +/- 0-1 (S.E.M.) i.u./l) increased to a maximum of 4-1 +/- 1-3 i.u./l (P less than 0-001) after 6 months, and FSH rose from 4-9 +/- 1-0 to a maximum of 7-7 +/- 1-3 i.u./l after 4 months of treatment (P less than 0-01). The response to luteinizing hormone releasing hormone (LH-RH) was studied at monthly intervals. Sums of increments of serum LH increased from 35 +/- 4 to 92 +/- 17 i.u./l at 4 months (P less than 0-001) and of FSH from 14 +/- 3-4 to 23 +/- 3-5 i.u./l at 4 months (P less than 0-01). Basal serum androgens rose from 25 +/- 2-7 to 38 +/- 2-4 nmol/l after 4 months of treatment (P less than 0-05), and serum oestradiol-17beta increased from 185 +/- 25 to 631 +/- 90 pmol/l by 6 months (P less than 0-001). No significant changes occurred in sperm counts. Five normal men acted as controls: they were given tamoxifen for 1 week. No significant changes were observed in serum LH, FSH or release of these hormones following administration of LH-RH. Serum androgens and oestrogens however, increased significantly by day 4 of treatment (P less than 0-05).


The hypothalamic centres involved in the control of production and release of prolactin in sheep. Lesions made in the anterior medial basal hypothalamus (MBH) in pregnant or lactating ewes caused lack of development of the mammary gland and depressed milk yield, a decrease in plasma prolactin concentration and structural changes in the prolactin cells as manifested by a lack of the expected degranulation (lack of the hormone release) or by atrophic changes and diminished cell granulation (or granule synthesis). Lesions made in the caudal MBH during pregnancy advanced development of the mammary gland, induced a rapid increase in the plasma prolactin concentration and caused extensive degranulation of prolactin cells. These results indicate that at least two functional systems controlling the secretion and production of prolactin exist in the hypothalamus of sheep: an anterior system which is stimulatory and a caudal which is inhibitory.


Effects of luteinizing hormone releasing hormone on plasma follicle-stimulating hormone and luteinizing hormone levels in the ferret. Heterologous radioimmunoassays for FSH and LH were employed to examine the effect of synthetic LH-RH upon gonadotrophin secretion in the ferret. Intravenous injection of 4 microng LH-RH induced a surge of FSH and of LH secretion in male and in female animals. In intact and in castrated males, the rise of LH was much more marked than that of FSH. The gonadotrophin response to LH-RH was greater in anoestrous than in oestrous females; FSH secretion was not enhanced during oestrus. Ovariectomized females behaved as anoestrous females with respect to LH secretion, while FSH secretion remained unchanged. Treatment of ovariectomized females with progesterone did not alter the pattern of response to LH-RH, but oestradiol treatment depressed the reaction to match that seen in oestrous females. Repetitive injections of LH-RH induced repetitive surges of FSH and LH in anoestrous females, but only of LH during oestrus: slow i.v. infusion of LH-RH induced a sustained elevation of plasma LH levels both in oestrous and in anoestrous females; again FSH levels rose only in anoestrous females. Injection of synthetic TRH did not alter gonadotrophin secretion in corresponding groups of male or female ferrets.


Evaluation of the technique of immunosurgery for the isolation of inner cell masses from mouse blastocysts. Inner cell masses (ICMs) immunosurgically-isolated from 31/2-day mouse blastocysts were examined for trophoblast cell contamination and developmental capacity. Blastocysts were preincubated in rabbit anti-mouse antiserum, washed thoroughly and then incubated in complement. The ICMs were then easily dissected by drawing through a fine pipette. Various experiments confirmed that the trophectoderm had been completely removed by this treatment. Firstly, the ICMs did not bind a fluorescein-conjugated antibody directed against rabbit IgG, indicating the absence of cells exposed to the rabbit antiserum during the immunosurgical procedure. Secondly, ICMs dissected from blastocysts preincubated in a suspension of melanin granules did not include any of the trophoblast cells that had phagocytosed the granules. And, thirdly, the protein synthetic profile of these ICMs was similar to microsurgically dissected ICMs, and in particular, trophoblast specific spots were absent. The developmental capacity of immunosurgically-isolated ICMs was tested by injecting them into blastocysts and transferring to the uterus of 2 1/2-day pseudopregnant recipients. Extensive chimaerism was detected in the majority of implants, 5-6 days after transfer, but only in ICM-derived tissues. This demonstrates both the lack of trophoblast cell contamination and functional viability of these ICMs.


The presence of two Fc receptors on mouse macrophages: evidence from a variant cell line and differential trypsin sensitivity. A stable variant of a clone of the P388D1 macrophage line was isolated using four cycles of treatment with mouse IgG2a-rabbit anti-kappa complexes and rabbit complement. The variant had the same Ka and about the same number of sites per cell for IgG2a as the parent line. However, the variant had 10% as many binding sites for rabbit IgG in soluble antigen-antibody complexes, and the affinity of binding was threefold higher. This change in binding of complexes to cells of a cloned line without alternation of IgG2a binding provides evidence for the presence of two distinct Fc receptors. The two receptors could also be distiguished on the P388D1 line and on thioglycollate-induced mouse peritoneal macrophages by differential sensitivity to trypsinization. The receptors that bind monomeric IgG2a, sheep erythrocytes (SRBC) covalently bound with IgG2a or rabbit IgG using glutaraldehyde, and Sephadex beads coupled with IgG2a or rabbit IgG using cyanogen bromide activitation, is sensitive to trypsinization. The receptor that binds soluble rabbit antibody-antigen complexes, trinitrophenyl-SRBC and dinitrophenyl(DNP)-bovine serum albumin Sephadex beads coated with rabbit anti-DNP IgG is trypsin resitant, the observation that uncomplexed rabbit IgG oes not bind to the trypsin-resistant receptor, whereas the same IgG bound to its antigen does, suggests that conformational changes induced by the binding of ligand may be of consequence in macrophage function.


Features of systemic lupus erythematosus in mice injected with bacterial lipopolysaccharides: identificantion of circulating DNA and renal localization of DNA-anti-DNA complexes. After injection of lipopolysaccharides (LPS) in mice, there is first a release of DNA into plasma and secondly an induction of anti-DNA antibodies. The circulating DNA was purified from plasma and physico-immunochemically characterized. This DNA has a similar density to mammalian cellular DNA,is 4--6S insize, and probably represents a mixture of single-stranded DNA (SSDNA) and double-stranded DNA (DSDNA) or DSDNA with some single-stranded regions. This purified DNA was shown to react with anti-DNA antibodies which appeared as early as 3 days after a single injection of LPS in mice. In serum, DNA-anti-DNA complexes were not detected, although unidentified circulating immune complex-like material was demonstrated 5-8 days after the injection of LPS. In tissues, particularly in renal glomeruli, fine granular immune complex-type immunoglobulin deposits appeared along the glomerular capillary walls and in the mesangium 3 days after the injection of LPS. There is a direct correlation between the level of anti-DNA antibodies and the intensity of glomerular deposits and about 40% of immunoglobulins eluted from kidneys are anti-DNA antibodies, indicating that some of the immune complexes localized in kidneys are DNA-anti-DNA complexes. Based on these observations, the following hypothetical mechanism for the glomerular localization of DNA-anti-DNA complexes after the injection of LPS in mice is proposed. First, DNA, which has been released in circulating blood after injection of LPS, might bind to renal glomeruli, probably on glomerular basement membranes (GBM) through a high affinity of GBM for DNA; secondly, circulating anti-DNA antibodies, which appear later, might react with the glomerular-bound DNA and form immune complexes independently of circulating immune complexes. However, the possibility of direct deposition of immune complexes is not ruled out.


Recirculating, suppressor T cells in transplantation tolerance. An adoptive transfer system was used to examine the capacity of cellular inocula from rats fully tolerant of Ag-B antigens to transfer tolerance to irradiated recipients. Permanent tolerance in these irradiated recipients involved specific suppression of the regenerating immune response. Cells obtained from tissues rich in recirculating lymphocytes were the most effective suppressors. Highly purified inocula of T cells from tolerant donors were potent suppressors in irradiated hosts, but were not capable of direct suppression of peripheral antigen-sensitive T cells.. The role of the thymus in maintaining the complement of recirculating suppressor T cells in tolerant animals was examined after adult thymectomy. Thymectomized tolerant rats did not reject their tolerated grafts, and the longevity of the suppression in tolerant rats was confirmed by showing that adoptive transfer of cells from thymectomized tolerant donors was effective in suppressing irradiated recipients up to 180 days after thymectomy. Cellular inocula from these donors appeared to lose their suppressor function marginally faster than they lost effector function (as measured by their capacity to mediate rejection of third party control grafts). Thymectomy made tolerant rats more vulnerable to the termination of tolerance by challenge with normal cells. Transplantation tolerance is maintained in adult rats by long-lived rapidly recirculating suppressor T cells. The target for the suppressor action of these cells is probably the precursor of alloantigen-sensitive lymphocytes, and the effect of suppression may be deletion or inactivation of the relevant clone of these cells.


The relationship between surface immunoglobulin isotype and immune function of murine B lymphocytes. I. Surface immunoglobulin isotypes on primed B cells in the spleen. We investigated the ability of IgM-, IgD-, and IgG-bearing cells from the spleens of (BALB/c x C57BL/Ka)F1 mice primed to dinitrophenyl-bovine serum albumin (DNP-BSA) to restore the adoptive secondary anti-BSA and anti-DNP antibody responses. A rabbit anti-mouse IgD antiserum was prepared and the specificity documented by radioimmunoprecipitation, and cell surface staining. Purified populations of IgM-, IgD-, and IgG-bearing cells were prepared by immunofluorescent staining with isotype-specific reagents, and sorting on the fluorescence activated cell sorter. Bright or dull cells were transferred to irradiated syngeneic recipients which were challenged with DNP-BSA in saline. Unfractionated spleen cells restored an adoptive secondary serum antibody response which was all IgG (2-mercaptoethanol resistant). Purified IgM- or IgD-bearing cells restored both the secondary IgM and IgG antibody response. IgG-bearing cells restored only the IgG response. In addition, the IgG-bearing cells appear to suppress the adoptive secondary IgM response, since depletion of IgG-bearing cells from transferred spleen cells results in a marked increase in the adoptive IgM response.


The relationship between surface immunoglobulin isotype and immune function of murine B lymphocytes II. Surface immunoglobulin isotopes on unprimed B cells in the spleen. We investigated the ability of IgM-, IgD-, and IgG-bearing cells from the spleens of unprimed (BALB/c x C57BL/Ka)F1 mice to restore the adoptive primary anti-BSA and anti-DNP antibody responses. Purified populations of isotype-specific cells were prepared by immunofluorescent staining and sorting on the fluorescence activated cell sorter. Bright or dull cells were transferred to irradiated syngeneic recipients which were challenged with DNP-BSA in complete Freund's adjuvant. Unfractionated spleen cells as well as IgM- and IgD-bearing cells restored the adoptive primary IgM and IgG antibody response. IgG-bearing cells restored a vigorous adoptive response which was all IgG (2-mercaptoethanol resistant). Depletion of IgG-bearing cells markedly increased the adoptive IgM response, and depletion of IgM-bearing cells markedly increased the IgG response. However, depletion of IgD-bearing cells resulted in a considerable reduction in the IgG response. The latter finding indicates that there is a subpopulation of IgD-bearing cells which express little or no surface IgM and which make a considerable contribution to the adoptive primary IgG response.


Mechanism of T-cell help in the immune response to soluble protein antigens. I. Evidence for in situ generation and action of T-cell-replacing factor during the anamnestic response to dinitrophenyl keyhole limpet hemocyanin in vitro. The involvement of a nonantigen-specific T-helper factor in the anamnestic immune response to dinitrophenyl keyhole limpet hemocyanin is demonstrated employing cultures of unseparated spleen cell populations. In such cultures of primed and boosted spleen cells, a good IgG anti-DNP response could be obtained if the hapten was presented on a heterologous carrier, provided that the homologous carrier was added simultaneously. T-cell depletion and reconstitution experiments show that such a factor, presumably identical with T-cell-replacing factor is produced by primed helper cells upon rechallenge and helps primed B cells, stimulated by soluble heterologous carrier hapten conjugates, to become IgG-secreting cells.


Mechanism of T-cell help in the immune response to soluble protein antigens II. Reconstitution of primary and secondary in vitro immune responses to dinitrophenyl-carrier conjugates by T-cell-replacing factor. Spleen cells of dinitrophenyl keyhole limpet hemocyanin (DNPKLH) primed and boosted mice produced a nonantigen-specific helper factor upon in vitro challenge with DNPKLH. This helper factor displays all of the biological characteristics so far described for TRF produced by allogeneic or Concanavalin A stimulation of mouse spleen cells. It restores the primary anti-SRBC response in nude spleen cultures following the same kinetics of action as T-cell-replacing factor (TRF). Conversely, TRF restores the primary in vitro immune response of nude spleen cultures to DNPKLH. TRF also restores the secondary anti-hapten IgG response of T-cell-deprived spleen cell cultures derived from DNPKLH primed and boosted mice. Here the need for carrier specificity is fully overcome. The data therefore suggest that TRF, as a nonantigen-specific maturation signal, is involved in the primary and secondary immune responses to both particulate and soluble antigens.


Induction of autoimmunity in good and poor responder mice with mouse thyroglobulin and lipopolysaccharide. The administration of soluble mouse thyroglobulin (MTg) in conjunction with bacterial lipopolysaccharide (LPS) led to the termination of natural tolerance to MTg in mice. The extent of autoimmunity correlated with responsiveness to MTg, previously shown by the injection of MTg in complete Freund's adjuvant (CFA) to be dependent upon the H-2 haplotype. In good responder B10.BR (H-2k) mice given MTg either with LPS or in CFA, high antibody levels to MTg and extensive mononuclear cell infiltration in the thyroid were observed. In contrast, congenic poor responder B10.D2 (H-2d) mice given MTg plus LPS showed low levels of antibody to MTg, compared to those receiving MTg in CFA, and insignificant cellular infiltration of the thyroid. In no instance did autoimmunity develop in either good or poor responder strain given MTg, LPS, or CFA along although LPS was antigenic in both of these congenic strains. Since the genetic difference in responsiveness to MTg is known to be T-cell based, the involvement of T cells in LPS-treated mice was suspected. This was further ascertained by the use of athymic poor responder (BALB/c) mice and thymectomized, irradiated, and bone marrow-reconstituted B10.BR mice. Antibodies to MTg were detected only in heterozygous (nu/+) mice and good responder mice reconstituted with both thymus and bone marrow cells. In addition, significant cellular infiltration in the thyroid occurred only in fully reconstituted good responder mice. Thus, the adjuvant effect of LPS on responsiveness to MTg required T cells. Since unmodified MTg and LPS abrogated selftolerance to MTg, the need for cross-reactive T cells could be excluded. These observations suggest the presence of self-reactive T cells.


Restriction of primary responses to the IgG class and dependency of IgM responses on secondary immunization for the copolymers of L-glutamic acid, L-tyrosine, and L-alanine. Primary responses to the linear polymers of L-glutamic acid, L-tyrosine, and L-alanine are restricted to the IgG class of antibodies. The appearance of specific IgM antibodies against these antigens is dependent upon secondary immunization, in contrast to many classical antigenic systems. The presence of an IgM response was verified by a direct plaque-forming cell assay, the inhibition of direct plaques by an antiserum specific for mouse micron-chain, and the physical separation of IgM and IgG GAT-specific antibodies by gel filtration. Preimmunization of the appropriate nonresponder strain with GAT or GT inhibits both the secondary IgM and IgG responses to GAT-MBSA and GT-MBSA, respectively. The tolerance observed is due to the induction of suppressor cells as demonstrated by cell transfer experiments.


[The mechanization of the enzyme immunological determination of phenytoine with the Centrifichem (author's transl)]. The separation of free from bound antigen is not required in homogeneous enzyme immunoassays (EMIT). Therefore this technique can easily be mechanized. For the mechanization of the enzyme immunological determination of serum levels of Phenytoine the fast analyzer (CENTRIFICHEM) was found most satisfactory in our laboratory. The marked advantages over manual methods are: 1. The amount of sample and reagents needed can be reduced by half. 2. The intra- and interassay variance are significantly better compared to values found with manual techniques. 3. 60 samples can be determined in duplicate in under two hours.


Redistribution of myosin accompanying capping of surface Ig. The capping of surface Ig on B cells occurs with a striking redistribution of cytoplasmic myosin. Our results suggest a close association between surface Ig and myosin which could be the basis for Ig redistribution and stimulated motility.


Epidermal-dermal tissue interactions between mutant foot skin and normal back skin: a comparison of the inductive capacities of scaleless low line and normal anterior foot dermis. The inductive capacities of 9- to 16-day anterior foot dermis of scaleless low line and normal embryos were compared by recombining them with a common source of epidermis, i.e., 7-day normal back epidermis. Tissue recombinants were cultured as grafts to the chorioallantoic membrane (CAM). Both normal and scaleless low line dermis of 12 to 13 days of incubation began to lose their ability to elicit feather production in 7-day normal back epidermis. Normal foot dermis began to elicit scale production at 12 to 13 days, whereas scaleless low line anterior foot dermis maintained feather production at a low level. It is inferred that without being associated with scale placode formation, scaleless low line anterior foot dermis does not acquire specific inductive capacities related to the production of an outer scale surface in the overlying epidermis. Feather placodes do not function as surrogates of scale placodes. The difference between normal and scaleless low line anterior foot dermis in terms of specific inductive capacities related to scale production is interpreted as a secondary effect of the action of the scaleless allele in interfering with scale placode formation in the scaleless low line anterior foot epidermis.


Immunofluorescent localization of acrosin in mammalian spermatozoa (1). Acrosin was detected by immunofluorescence in the spermatozoan acrosomes of artiodactyla (bull, ram and boar), perissodactyla (horse), carnivora (dog and cat), lagomorpha (rabbit) and primates (human) using anti-bovine acrosin immunoglobulins. The results indicate that the acrosin molecules of several mammalian species possess antigenic similarities.


The enzymic degradation of an alkali-soluble glucan from the cell walls of Saccharomyces cerevisiae. An alkali-soluble glucan from the cell walls of Saccharomyces cerevisiae NCYC1109 has been hydrolysed with a purified endo-(1 leads to 3)-beta-D-glucanase and an endo-(1 leads to 6)-beta-D-glucanase from Bacillus circulans WL-12. The products of enzyme action include various oligosaccharide and polysaccharide fractions which have been separated by gel filtration and characterized, giving new information on the fine structure of the glucan. The isolated cell walls have also been subjected to enzymic hydrolysis. The results suggest that part of the cell-wall mannan is held in place by a glucan component.


Monitoring enzyme synthesis as a means of studying peptide transport and utilization in Escherichia coli. A new method has been developed for measuring peptide transport in aminoacid auxotrophs of Escherichia coli by following induction of beta-galactosidase. Appearance of the enzyme was determined after addition of inducer and peptides to amino-acid starved bacteria. For a given number of lysine equivalents, the rate and the extent of enzyme synthesis were the same for lysine and lysyl peptides; similar results were found for glycine and glycl peptides. Saturation constants for peptide transport were determined from the exogenous peptide concentration that gave half maximal rates of enzyme synthesis. The saturation constants, studies with mutants defective in peptide transport, and detection of competition between peptides for uptake, all endorsed earlier conclusions from growth tests about the structural specificities for peptide transport. The new method is quicker, more sensitive and more informative than growth tests.


Expression of Klebsiella pneumoniae nitrogen fixation genes in nitrate reductase mutants of Escherichia coli. Nitrate reductase (nar) A, B and E mutants of Escherichia coli with plasmids carrying Klebsiella pneumoniae nitrogen fixation (nif) genes reduced acetylene independently of added molybdate, but nar D mutants showed pleiotropic dependence on the concentration of added molybdate for expression of both nar and nif. No complementation of nar mutations by nif occurred; nitrite but not nitrate repressed nif in nar hosts. Derepression of nif occurred in molybdenum-deficient nar D (nif) strains since nitrogenase peptides were present. nifB mutants, thought to have a lesion in the pathway of molybdenum to nitrogenase, as well as nif deletion mutants, had normal nitrate reductase activity.


A novel pleiotropic mutation in Escherichia coli K12 which affects transduction, transformation and rates of mutation. A mutant strain of Escherichia coli K12, R2721, has been shown to differ from its parent strain, S491, in four associated phenotypic characters as a result of a single mutation. This strain did not give recombinants with DNA transduced by bacteriophage PI or bacteriophage Mu, nor transformats after exposure to R factor DNA: lysates of bacteriophage PI grown on this strain did not appear to contain any transducing particles when tested on normal recipients. Moreover, the reversion rates, both spontaneous and ultraviolet-induced, for two auxotrophic markers were reduced. The frequency of revertants was at least two orders of magnitude lower in cultures of R2721 than in cultures of S491I. Many of the rare revertants for one or other of the auxotrophic markers were found to have regained normal reversion frequencies for the other marker and for the capacity to be transduced. In all other respects, recombination in R2721 appeared normal, the frequency of chromosomal mobilization by and F' factor was unaffected and normal yields of recombinants were obtained from matings with Hfr strains. The only circumstance in which transduction of R2721 was observed was when the capacity to ferment galactose was selected and PI had been grown on a strain carrying lambdadgal when, presumably, integration was effected by the phage-coded gene products. The mutation has been located on the E. coli chromosone map between tonA and pro and has been given the symbol tdi (transduction inhibition). Double mutants, (tdi recA) and (tdi recB), have been isolated and show no unexpected properties.


Characteristics of the microplate method of enzyme-linked immunosorbent assay for the detection of plant viruses. Some characteristics of a microplate method for the detection and assay of plant viruses using enzyme-labelled antibodies are described. The method enabled the highly sensitive detection of a number of morphologically different viruses in purified preparations and in unclarified extracts of herbaceous hosts and of infected crop plants. Virus concentrations were estimated by photometric measurement of the colour intensity of the hydrolysed substrate. The suitability of the tehcnique for various field and research applications is considered.


Demonstration of nuclear immunofluorescence in Rift Valley fever infected cells. Eosinophilic intranuclear filaments described previously in Rift Valley Fever infected cells, were shown to fluoresce specifically in an indirect technique with antiserum to the virus. Actinomycin D failed to suppress development of the filaments or replication of the virus.


Jewish culture and psychological differentiation--partial replication. A replication study was made to test the following hypotheses: (a) Traditional-Jewish Ss, in comparison with Anglo-Saxon Protestant Ss, matched for age and verbal ability, will be more global on criterion measures of psychological differentiation. (b) Within group intercorrelations of these measures will all be in the positive direction. Ten pairs of boys (ages 9-11) were compared on the Embedded Figures, Human Figure Drawing, and Verbal Disembedding tests. The first hypothesis was confirmed with regard to Embedded Figures (p less than .05), but not with regard to the other two criterion measures. The within group intercorrelations were r = .25 to .48 for Embedded Figures versus the other two measures. The correlations between Verbal Disembedding and Human Figure Drawing were small and negative (all correlations were nonsignificant).


Alterations in forebrain catecholamine metabolism produced by cerebellar lesions in the rat. Projections from the midline cerebellar nuclei to norepinephrine (NE) and dopamine (DA) cell groups in the brain stem have been demonstrated histologically. To determine if these connections are significant biochemically, unilateral electrolytic lesions were placed in either vermis or paravermis and levels of DA, NE and gamma-aminobutyric acid (GABA) were measured in each half of the forebrain at 1 1/2, 3 or 6 weeks. In the cerebral hemisphere ipsilateral to a vermis lesion, there was a decrease in NE levels at 3 and 6 weeks. Relative to the opposite side DA was also reduced at 3 and 6 weeks. Paravermal lesions caused a contralateral reduction in DA at 3 weeks but no change in NE. GABA was only slightly altered. These results suggest that the cerebellum can modify levels and turnover of catecholamines in the brain, possibly via direct anatomic connections as well as by functional interaction with catecholaminergic pathways.


Interaction of haloperidol and gamma-butyrolactone with (+)-amphetamine-induced changes in monoamine synthesis and metabolism in rat brain. The interaction of (+)-amphetamine with haloperidol and gamma-butyrolactone on synthesis of monoamines in rat brain regions was investigated using an in vivo method, in which the accumulation of dopa and 5-hydroxytryptophan (5-HTP) was measured after inhibition of the aromatic amino acid decarboxylase by means of 3-hydroxybenzylhydrazine. The accumulation of 3-methoxytyramine (3-MT) after inhibition of the monoamine oxidase with pargyline was taken as an indicator of the in vivo release of dopamine (DA) into the extraneuronal space. (+)-Amphetamine at doses of 1--3 mg/kg caused an increase of dopa formation in the DA-rich areas c. striatum and mesolimbic forebrain but had no effect on dopa formation in neocortex and 5-HTP formation in all brain regions investigated. At a dose of 10 mg/kg (+)-amphetamine decreased dopa as well as 5-HTP formation in all brain regions studied. 3-MT accumulation in whole rat brain was stimulated by (+)-amphetamine as well as haloperidol and inhibited by gamma-butyrolactone. Combined treatment with haloperidol and (+)-amphetamine not only potentiated the stimulation of dopa formation but also the increase of 3-MT accumulation. Pretreatment with gamma-butyrolactone antagonized the stimulation of 3-MT formation induced by (+)-amphetamine at a dose of 10 mg/kg. This dose of (+)-amphetamine markedly counteracted the gamma-butyrolactone-induced increase in dopa formation especially in the DA-rich areas. The data support the view that impulse flow in DA neurons facilitates the effect of (+)-amphetamine on DA release and DA synthesis. Inhibition of catecholamine synthesis after high doses of (+)-amphetamine may be due to an increase in cytoplasmatic DA concentration causing end-product inhibition of tyrosine hydroxylase.


6-hydroxydopamine destruction of central adrenergic neurones prevents or reverses developing DOCA-salt hypertension in rats. The role of brain catecholaminergic neurones in the pathogenesis of DOCA-salt hypertension in the rat was investigated by selective depletion of central catecholamines using intraventricular or intracisternal administration of 6-hydroxydopamine (6-OHDA). Only the intraventricular injections prevented the development of hypertension. In addition, intraventricular 6-OHDA reversed the hypertension produced by two weeks but not six weeks of DOCA-salt treatment. The ability of intraventricular injections of 6-OHDA to prevent or reverse DOCA-salt hypertension while intracisternal injections do not, appears to be related to the greater depletion of brain catecholamines produced by the intraventricular injections. Only in the spinal cord and in the locus coeruleus were the norepinephrine contents depleted equally by either injection route. These findings suggest that central catecholaminergic neurones other than those originating in the locus coeruleus or descending in the spinal cord are important in the initiation, but not in the long term maintenance, of DOCA-salt hypertension. The influence of the central catecholamine neurons involved in the development of DOCA-salt hypertension might be mediated neurally via nonadrenergic pathways or hormonally via the brain-pituitary-endocrine system.


Effect of apomorphine on schizophrenic symptoms. The effects of apomorphine on psychotic symptoms were evaluated in chronic schizophrenic patients using double-blind placebo controlled procedures. Although on the basis of dopamine theory of schizophrenia, apomorphine was expected to increase schizophrenic symptoms, in this study apomorphine substantially reduced psychotic symptoms in some chronic schizophrenic patients. No patient showed the substantial increase in psychotic symptoms previously demonstrated after the administration of IV methylphenidate. These clinical effects of apomorphine in schizophrenia may be relevant to recent pharmacological research which has indicated that apomorphine also has potent effect on presynaptic dopamine neurons, in addition to its previously described postsynaptic receptor stimulation.


Agonist--antagonist interaction on dopamine receptors in brain, as reflected in the rates of tyrosine and tryptophan hydroxylation. The effect of haloperidol and apomorphine, and both drugs in combination, on the first steps in the synthesis of catecholamines and 5-hydroxytryptamine (5-HT) has been studied in three rat brain regions. The rate of formation of dopa and 5-hydroxytryptophan (5-HTP) was studied by measuring the accumulation of these amino acids during 30 min after administration of the inhibitor of the aromatic L-amino acid decarboxylase, NSD 1015 (3-hydroxybenzylhydrazine HCl). Haloperidol caused an increase in dopa and no change in 5-HTP formation. The threshold dose was severalfold higher in the noradrenaline-predominated hemisphere portion than in the dopamine-rich striatal and limbic regions, suggesting a higher affinity of haloperidol for dopamine than for noradrenaline receptors. Apomorphine caused a decrease in dopa formation in all three brain regions studied, although the effect was much more pronounced in the regions dominated by dopamine. The threshold dose was about 30 microng/kg, i.e. an order of magnitude lower than the threshold dose for apparent postsynaptic dopaminergic receptor activation. This discrepancy is suggested to be due to preferential activation of inhibitory dopaminergic autoreceptors by low apomorphine doses. This phenomenon may also contribute to explain the complex dose-response curves of apomorphine. Low doses of apomorphine caused a decrease and high doses an increase in 5-HTP formation. These effects, like those on noradrenaline synthesis, are suggested to be secondary to activation of dopaminergic pre- and post-synaptic receptors. The interaction between apomorphine and haloperidol with respect to dopa formation appears to be largely explicable on the assumption of a competition between an agonist and an antagonist for dopaminergic receptors. However, very large doses of apomorphine cause a haloperidol-resistant inhibition of tyrosine, and probably also tryptophan, hydroxylation, which may be due to a direct inhibition of the aromatic amino acid hydroxylase involved.


A program to compute the sizes and numbers of spherical bodies from observations made on tissue sections. A computer program, written in standard Fortran, is given to facilitate the computation of the sizes and correct distribution of spheres from the number and size of the circular profiles observed in tissue sections. The method is particularly valuable in cases where it is necessary to determine absolute numbers of cells or subcellular fragments where the objects of interest are not of uniform size. The method gives true cell numbers and the distribution of the spherical diameters.


Interstitial gamma irradiation by 198Au of the pituitary in diabetic retinopathy. Selective growth hormone blockade and ocular results. Pituitary interstitial irradiation by stereotaxic implantation of a gamma pointemitter was performed in 110 patients with progressing diabetic retinopathy. Reduction of growth hormone secretion peaks was delayed, but was consistent and selective in all cases controlled by somatotropin stimulation test, while other pituitary secretions remianed unchanged in all but five cases. In 31 patients who had no associated photocoagulation, long-term postoperative assessment showed significant improvement of hemorrhages and intrarentinal new vessels with preservation of visual acuity. Preretinal vascular proliferation (except when treatable by photocoagulation), extensive retinitis proliferans, and macular deterioration are contraindications. This technique represents a new approach in the functional pituitary neurosurgical treatment of diabetic retinopathy and differs from all other procedures that result in a more or less complete hypophysectomy.


Effect of fat intake on cholesterol turnover and bile acid formation. Three groups of male rats were maintained on diets containing different amounts of fat. After one week on such regimens, they were injected intraperitoneally with cholesterol-3H. During the following 28 days, the radioactivity and quantity of fecal cholesterol and its metabolities were determined. The coprostanol excretion was the about same in all groups and the bile acids excretion increased with increasing fat intake; however, compared to the fat-free group, the excretion of the injected cholesterol-3H was greater in the 3% fat group and less in the 10% fat group. Consequently, the specific radioactivity of bile acids was lower in the 10% group than in the others. The half-life of labelled cholesterol was 12.6, 16.0 and 22.2 days for the 3%, 10% fat and fat-free groups, respectively. Rather than a fat-free diet, a low-fat diet of 3% or so, would be of more advantage in eliminating cholesterol by increasing the formation of bile acids to emulsify the fat. In the 10% fat group, however, the enlarged pool size of bile acid probably delayed cholesterol metabolism to bile acids.


Uptake and utilization of vitamin B6 and its phosphate esters by Escherichia coli. Escherichia coli KG980, a vitamin B6 auxotroph derived from E. coli K12, utilized the three unphosphorylated forms of vitamin B6, more or less effectively, for growth, but did not utilize the three phosphate forms at concentrations ranging from 10(-7) to 10(-5) M in the minimum medium of Davis and Mingioli. The bacterium, however, utilized the phosphate forms, although less effectively than the unphosphorylated forms, in the phosphate starving medium of Garen and Levinthal which is known to derepress alkaline phosphatase. Correspondingly, the phosphate forms of 3H-labeled vitamin B6 in the minimum medium were not taken up by the bacterial cells grown in the same medium, but were taken up when the phosphate starving medium was used for growth and uptake experiments; the unphosphorylated forms were taken up with either of the media used. After 30-min incubation of the cells grown in the phosphate starving medium with 3H-pyridoxine phosphate added to the same medium, the main extracellular 3H-vitamin B6 was found to be pyridoxine, evidently indicating an involvement of alkaline phosphatase action. It is concluded from these results that the phosphate forms of vitamin B6 can be taken up and utilized only after dephosphorylation but not taken up in their intact form. The initial rate of 3H-pyridoxal and 3H-pyridoxamine uptake in the minimum medium showed saturation kinetics. The Km and Vmax values were 1.2 X 10(-6) M and 62 pmoles/min/mg (dry cell weight) for pyridoxal; 11 X 10(-6) M and 65 pmoles/min/mg for pyridoxamine. 3H-Pyridoxine uptake apparently consisted of a high-affinity saturable component, whose Km value was tentatively estimated to be 2.2 X 10(-6) M, and an unsaturable component. The uptake rates of these three unphosphorylated vitamin B6 compounds compared at limiting concentrations for the growth of E. coli KG980 appear to be essentially in good agreement with the response pattern of this bacterium to the three compounds.


Absence of health hazards associated with RDX manufacture and use. A cross-sectional spidemiologic study was conducted to investigate a cluster of three cases of systemic lupus erythematosus at one munitions plant. The study demonstrated no excess of autoimmune disease and also failed to identify any abnormalities of the hematologic, hepatic, or renal systems in employees with 8 hour time-weighted exposures to RDX of up to 1.57 mg/m3 (average exposure was 0.28 mg/m3).


The periodontium and restorative procedures. A critical review. The literature associated with the effect on the periodontal tissues of restorative procedures has been reviewed. Opinions have changed from the assumption that only poor restorative dental treatment led to periodontal disease, to the present concept, that even clinically sound restorations, if positioned subgingivally, may be important aetiological factors in the initiation of periodontal disease.


The influence of the time of etching and washing on the bond strength of fissure sealants applied to enamel. Variation of the etching and washing times of enamel surfaces prior to the application of fissure sealants affect the bond strength in differing ways. Depending on the length of the etch time, the effect of washing may increase or decrease the bond strength of the sealant. Clinically, the most satisfactory combination of etch and wash time was found to be different to that suggested by the manufacturers.


Polymer adhesion to enamel. The influence of viscosity and penetration. For a four-component acrylate adhesive system the influence of viscosity and penetration on adhesion to enamel was investigated. While low viscous monomer adhesives yield both inter- and intraprismatic penetration in enamel after polymerization, highly viscous adhesives result in the former type only. This difference in penetration of the adhesive in enamel is reflected quantitatively in the tensile strength values. Scanning electron microscopic measurements on the tag formation under various circumstances have been carried out. The SEM experiments show, e.g. that at polymerization times less than 1-5-2 min (a clinical necessity) polymer-tags were usually fractured from the bulk material due to stresses generated.


Treatment of the oral complications of leukemia. Complicating infectious processes and manifestations of immunosuppression were either cured or symptoms were decreased with aggressive therapy in 24 patients with acute leukemia and five with chronic leukemia. Oral surgical intervention combined with histocompatability-matched transfusion therapy can safely eliminate sources of sepsis and alleviate pain in patients who have undergone immunosuppressive and myelosuppressive treatment.


Life-threatening swelling after mandibular vestibuloplasty. This case report illustrates the potential for severe hemorrhage which authors have alluded to in articles on the vestibuloplasty procedure. The surgeon must be prepared to deal not only with the hemorrhage, but also with the sudden and possible life-threatening swelling that may accompany the bleeding.


Pancreatic alpha and beta cell functions in cystic fibrosis. Insulin and glucagon secretions were studied during oral glucose tolerance testing and arginine infusion in 13 patients with cystic fibrosis. Two groups of patients were identified; Group I (N=6) whose OGTT was entirely normal and Group II (N=7) who had some abnormality in glucose during OGTT. In each group basal glucagon concentrations were normal and supressed appropriately (p less than 0.05) after glucose; insulin responses were attenuated and the peak responses delayed. During arginine stimulation, insulin secretion was impaired in each group. However, glucagon secretion was diminished only in Group II. Thus, insulinopenia was found in both groups and hyperglucagonemia was not found as a contributory factor to the hyperglycemia in Group II.


The effects of single infusion of hypertonic sodium bicarbonate on body composition in neonates with acidosis. Acid-base equilibrium and plasma and red blood cell water and solute were evaluated in a group of asphyxiated, acidotic neonates prior to and following infusion of hypertonic NaHCO3. The dose was calculated to correct the deficit of base in a bicarbonate space of 400 ml/kg and was given at a rate of 0.3 mM NaHCO3/kg/minute. All of the infants with RDS and two of the five with other forms of asphyxia received ventilatory assistance during the infusion. The quantity of base infused was sufficient to alter acid-base balance and shift whole blood and red blood cell pH values toward normal. The changes in body composition 3 minutes following the infusion indicate that the osmotic load imposed by the hypertonic NaHCO3 caused a shift of solute-free water into the interstitial and intravascular fluids. During the period from 3 to 30 minutes following the infusion there was redistribution of extracellular water and solute so that plasma volume and [Na]PL decreased. Since there was no evidence of an intracellular shift of solute, we hypothesize that the changes in body composition between 3 and 30 minutes postinfusion were in part the consequence of gradual penetration of transcellular fluids by Na. Osmotic inactivation of ECF Na by sequestration with connective tissue polyelectrolytes may also play a role. These studies' do not provide an answer to the clinical problem of whether the beneficial effects of prompt correction metabolic acidosis outweigh the potenially harmful effect of the osmotic alterations that accompany rapid infusion of hypertonic NaHCO3.


Hypotension and bradycardia associated with airblock in the neonate. Preliminary report. The hospital courses of 19 infants were analyzed retrospectively to determine if there was a relationship between hypotension and extra-alveolar air: interstitial emphysema, pneumomediastinum, and penumothorax. Of the 16 infants who subsequently developed PT, 12 had hypotension with IE/PM from one to 86 hours prior to the onset of PT (median 6.5 hours). When compared to blood pressure values obtained before the development of IE/PM, the decrease was statistically significant (p less than 0.001). Four of the six infants of greater than or equal to 38 weeks' gestation had bradycardia throughout the duration of EAA. Hypotension and bradycardia appear to be associated with airblock and thus may alert the physician to those infants who are at greatest risk for developing PT.


Minoxidil therapy in children with severe hypertension. Six children, from 1.3 to 18 years of age, with severe hypertension associated with the hemolytic uremic syndrome, periarteritis, and renal transplant rejection received minoxidil, an antihypertensive agent, for three to 36 weeks. All had severe hypertension resistant to oral antihypertensive medications; five required frequent intravenous diazoxide therapy prior to minoxidil therapy. The mean pretreatment systolic and diastolic blood pressures were 176 and 117 mm Hg, respectively. Following treatment, the mean systolic and diastolic blood pressures were 133 and 82 mm Hg, respectively. Concomitant antihypertensive medications were decreased in all six patients once optimal blood pressure control was obtained. The initial dosage of minoxidil was 0.1 to 0.2 mg/kg/day; maximal dosage for blood pressure was 0.3 to 1.4 mg/kh/day. Major complications of therapy were fluid retention and hirsutism. Transient asymptomatic pericardial effusions occurred in two patients. Three patients on prolonged minoxidil therapy had persistent increases in right ventricular end diastolic diameters. Minoxidil is an effective oral antihypertensive agent for treatment of severe hypertension in pediatric patients. Avoidance of fluid retention is mandatory to prevent congestive heart failure.


Use of nalotone to to reverse narcotic respiratory depression in the newborn infant. Twenty neonates whose mothers had received meperidine (1.0 to 1.5 mg/kg) intravenously within three hours of delivery were studied to determine the effectiveness of naloxone in reversing neonatal respiratory depression. The following measurements were carried out within 20 to 30 minutes after delivery: minute ventilation, end tidal CO2, and ventilatory response to CO2. These determinations were repeated after administration of either placebo or naloxone, 0.01 mg/kg intramuscularly. Minute ventilation and PAco were within a normal range before medication in both groups, but the slope of the CO2 response curve was decreased, indicating mild-to-moderate respiratory depression. After administration of placebo the test results did not change significantly. After administration of naloxone, VE increased significantly (P less than 0.05) and the slope of the CO2 response curve doubled (P less than  0.001). Naloxone effectively reverses narcotic depression of the respiratory center in the newborn infant.