Action of ionophore A23187 at the cellular level. Separation of effects at the plasma and mitochondrial membranes. Bovine epididymal spermatozoa incubated aerobically in vitro in the presence of 0.1 to 0.2 mM CaCl2 accumulate 25 to 50 nmol of calcium/10(8) cells. The addition of low concentrations of the ionophore A23187 (0.01 to 0.5 nmol/mg of sperm protein) induces efflux of this accumulated calcium. At high ionophore concentrations (0.5 to 5.0 nmol/mg of sperm protein), calcium release is followed by an influx of up to 25 nmol of calcium/10(8) cells that is not dependent on mitochondrial energization. A selective increase in the permeability of the sperm plasma membrane produced by treatment with the polyene antibiotic, filipin, results in the release of that calcium which is accumulated in the presence of high concentrations of A23187. Sperm first treated with filipin possess the ability to accumulate and retain calcium (in the presence of an oxidizable substrate) but release Ca2+ without subsequent reaccumulation after the addition of 3 nmol of A23187/mg of protein. These observations are explained by the existence of competing calcium pumps operating within the mitochondrial and plasma membranes of the spermatozoan. Treatment with high concentrations of A23187 allows calcium influx into a non-mitochondrial compartment of the sperm cell as a consequence of the equilibration of this cation across both mitochondrial and plasma membranes. The amount of calcium uptake and its sensitivity to filipin indicate that calcium binding to soluble, intracellular components is also involved. The ability of low concentrations of A23187 to induce calcium efflux is explained as a result of the continued operation of the plasma membrane pump coincident with ionophore-induced decay of the concentration gradient across the mitochondrial membrane. This hypothetical action of low levels of the ionophore on the mitochondria is supported by the observation of net movements of calcium with filipin-treated cells and the respiratory responses and movements of phosphate and membrane-associated calcium with intact sperm. It is suggested that the basis of this apparent selectivity of ionophore action lies in the relative activities and kinetic properties of the competing calcium pumps in the plasma and mitochondrial membranes of these cells. Ionophore-induced influx of calcium into the extramitochondrial space results in a stimulation of respiration and kinetic activity of the sperm. This activation of motility is observed also with cells made entirely dependent upon glycolysis (by treatment with respiratory inhibitors) and suggests a direct involvement of calcium in the regulation of flagellar function.


Biochemical characterization of mutant forms of DNA polymerase I from Escherichia coli. II. The polAex1 mutation. DNA polymerase I has been purified to homogeneity from an Escherichia coli K12 strain bearing the temperature-sensitive conditionally lethal mutation, polAex1. The purified enzyme shows no defect in its polymerase or 3' leads to 5'-exonuclease activities; however, its 5' leads to 3'-exonuclease activity is abnormally low at both 30 degrees and 43 degrees. Although the mutant enzyme is able to catalyze the coordinated 5' leads to 3' polymerization and 5' leads to 3' exonucleolytic hydrolysis of nucleotides at a nick in duplex DNA ("nick translation") at a measurable rate at 30 degrees, this reaction is undetectable at 43 degrees. This defect is very likely responsible for the retarded joining of nascent DNA fragments and the consequent loss of viability that occur in the mutant at this temperature.


On the processive mechanism of Escherichia coli DNA polymerase I. Delayed initiation of polymerization. Escherichia coli DNA polymerase I shows a delay in the initiation of polymerization after binding to the cohesive ends of lambda DNA. This delay is significantly longer than the time required for the synthesis of an octanucleotide sequence on the right-hand cohesive end of the lambda DNA. When the extent of polymerization is limited by omission of one or more of the deoxynucleoside triphosphates, and polymerization started again by their addition, the delay still occurs. A plausible explanation for this phenomenon is that two forms of the enzyme or enzyme - DNA complex exist, only one of which is active. The delay, therefore, represents the time necessary to convert the inactive to the active form of the enzyme or enzyme complex. One consequence of the defect in DNA polymerase I, due to the polA12 mutation, is apparently to alter the equilibrium between the two forms. However, the rate of polymerization and the rate of conversion of inactive to active enzyme or enzyme complex are not changed significantly.


Identification of sn-glycero-1-phosphate and phosphoethanolamine residues linked to the membrane-derived Oligosaccharides of Escherichia coli. A previous report from this laboratory (van Golde, L.M.G., Schulman, H., and Kennedy, E.P. (1973) Proc. Natl. Acad. Sci. U.S.A. 70, 1368-1372) described the discovery in Escherichia coli of a novel class of oligosaccharides, containing glucose as the sole sugar, substituted with glycerophosphate units derived from membrane phospholipids, and with succinic acid in O-ester linkage. These membrane-derived oligosaccharides, comprising about 0.5 to 1.0% of the dry weight of E. coli, represent a family of closely related oligosaccharides that may be subfractionated on anion exchange resins. The present paper describes studies of the oligosaccharide A-1 described by van Golde et al. in the previous report. The glycerophosphate linked to the oligosaccharide in phosphodiester bond is the sn-glycero-1-P enantiomer. This finding strongly supports the previous conclusion that the oligosaccharides are the acceptors of the polar headgroups of membrane phospholipids, since the unesterified glycerophosphate of phosphatidyl glycerol is an sn-glycero-1-P residue, otherwise rare in nature. The glycerophosphate residues in the membrane-derived oligosaccharide are not substituted in the sn-2 or sn-3 positions, since they are readily oxidized by periodate under mild conditions. Alkaline hydrolysis liberates glycerophosphate, and only negligible amounts of free glycerol, consistent with the view that the glycerophosphate residues are linked to glucose units through position 6, unfavorable for the formation of glucose cyclic phosphate intermediates that would eliminate free glycerol. Oligosaccharide A-1 (but not Fraction A-2) contains phosphoethanolamine residues equivalent to 30 to 40% of the total phosphorus. The phosphoethanolamine residues are linked to position 6 of glucose units, as proved by the isolation of glucose 6-phosphate as a product of partial acid hydrolysis.


Oxidation of the carbanion intermediate of transaldolase by hexacyanoferrate (III). The transaldolase-dihydroxyacetone carbanion intermediate formed in the reaction of transaldolase with its donor substrates fructose-6-P or sedoheptulose-7-P is susceptible to oxidation by hexacyanoferrate(III). The dihydroxyacetone moiety is oxidized to the corresponding 2-ketoaldehyde, i.e. hydroxy-pyruvaldehyde (CH2OH-CO-CHO). This oxidation product is, in contrast to dihydroxyacetone, readily released from the enzyme. In the presence of hexacyanoferrate(III) transaldolase thus functions as an efficient catalyst of the oxidative cleavage of its donor substrates fructose-6-P or sedoheptulose-7-P into hydroxypyruvaldehyde and glyceraldehyde-3-P or erythrose-4-P, respectively. Two moles of hexacyanoferrate(III) are reduced per mole of oxidatively cleaved donor substrate. The molecular activity for oxidative cleavage of fructose-6-P at a hexacyanoferrate(III) concentration of 0.5 mM is 0.65% of that for the normal transfer reaction with erythrose-4-P as the acceptor substrate. The present data emphasize the applicability of certain oxidants as trapping agents for enzymatic carbanion intermediates as proposed previously (Healy, M.J.,, and Christen, P. (1973) Biochemistry 12, 35-41).


Solitary unicameral bone cyst: treatment with freeze-dried crushed cortical-bone allograft. A review of one hundred and forty-four cases. One hundred and forty-four solitary bone cysts were treated by curettage and packing with freeze-dried crushed cortical-bone allograft. One hundred and eight healed primarily. There was a higher rate of recurrence in young patients (less than ten years old), in active cysts, in females, and in incompletely packed cysts. However, of the cysts that were completely packed, 88 per cent healed. These data show that freeze-dried allogeneic crushed cortical bone is superior to similiarly processed cancellous bone and gives results similar to those achieved with fresh autogenous cancellous bone. Orthopaedic surgeons should therefore consider the use of freeze-dried allogeneic crushed cortical bone instead of autogenous grafts to avoid the morbidity and increased risk of complications associated with the procedure to obtain the autogenous bone.


Posterior bone-grafting for infected ununited fracture of the tibia. Twenty-three patients with severe open tibial fractures complicated by established deep infection and delay or failure of union were treated with posterior tibial bone-grafting. This was followed by immediate ambulation in a plaster cast. Tibial union was achieved in all twenty-three patients. Drainage ceased prior to or simultaneously with union in twenty-one patients, including twelve of fourteen patients who had indolent drainage from the anterior compartment of the leg at the time of posterior tibial bone-grafting.


Restoration of muscle balance of the foot by transfer of the tibialis posterior. A new technique of tibialis posterior transfer is described which has been used in a wide variety of conditions producing muscular imbalance in the foot. The results in eighty-five feet are reviewed in terms of range of motion, power and voluntary control of the transfer. The effect of the transfer on shoe wear, on the necessity for bracing and on the child's or his parents' assessment of the results are used to allocate an overall evaluation of the operation. Recommendations on the indications for the operation are given.


Factors in the adherence of flexor tendon after repair: an experimental study in the rabbit. The effects of splintage, suture and excision of the tendon sheath on the healing of incompletely transected flexor tendons in the rabbit have been evaluated separately and in various combinations. When all procedures were done together, repair was accompanied by dense adhesion formation with little evidence of any healing activity by the tendon cells. The experiments indicated that the adhesions were the result not of any one single factor studied but of all three contributing in varying degrees. Suturing produced the most adhesions but synovial sheath excision and immobilisation also contributed. It is suggested that these factors are also responsible for the adhesions which occur after flexor tendon repair in clinical practice.


Subcellular localization of diffusible ions in the yeast Saccharomyces cerevisiae: quantitative microprobe analysis of thin freeze-dried sections. In yeast cells, of which the intracellular potassium had been partly replaced by rubidium or caesium, the intracellular ion distribution was studied by means of energy-dispersive X-ray microanalysis. The cells were rapidly frozen and thin sections were cut at low temperature on a cryo-ultramicrotome without the use of a trough liquid. By this dry cryosectioning procedure, complete retention of the diffusbile ions in the cells was obtained. Unless the sections had been exposed to moisture, no signs of redistribution were apparent. For quantitative determinations a gelatin standard, containing known amounts of the elements of interest, was prepared in the same way as the cells. The concentrations of potassium, rubidium, caesium and chloride in the nucleus, the cytoplasm and the vacuole could be measured. The intracellular distributions of potassium, rubidium and caesium were very similar. The concentrations of these ions in the cytoplasm were about equal to those in the nucleus and twice those in the vacuole. The total concentration in the cytoplasm was 180-190 mmol/kg fresh weight, in the nucleus 190-200 mmol/kg fresh weight and in the vacuole 75-90 mmol/kg fresh weight. The permeability of the yeast cell for chloride is markedly lower than for the cations.


Cell organelles at uncoated cryofractured surfaces as viewed with the scanning electron microscope. A method of direct visualization of cell organelles by scanning electron microscopy (SEM) is described. Plant and animal tissues fixed in glutaraldehyde and osmium tetroxide are treated with the ligand thiocarbohydrazide and a second osmium tetroxide solution, to increase their osmium content. Tissues are then dehydrated, infiltrated with an epoxy monomer, and together solidified with dry ice and fractured. The pieces are transferred to pure acetone, critical-point dried, attached to stubs with silver paint and viewed by SEM. The ligating procedure increases the osmium concentration at its original bonding site sufficiently to render the tissue electrically conductive, thus obviating the need for metallic coating. he organelles at the fractured surface are revaled in relation to their osmium incorporation rather than by surface irregularities as with coating methods. The image derived from the uncoated surface approaches in resolution that of transmission electron micrographs of thin sections. A protion of the image arising from a small distance below the surface, while at progressively lower resolution, provides some 3-dimensional information about cell fine structure.


[Intraepiploic pancreatic autograft in dogs. Blood sugar changes as compared with a group of pancreatectomized dogs]. The authors used the omentum to envelop thin pancreatic fragments (10 to 18 g. i.e. 1/3 to 1/2 of the canine pancreas). After control of the blood sugar, a total corporo-caudal pancreatectomy was carried out in 21 dogs within 1 to 5 months. Nine survivors from one week to 3 months, permitted them to control the course of the blood sugar with biopsies of the graft and the absence of residual pancreas in the duodeno-hepatic region at autopsy. Compared with a comparative series of 11 dogs which survived total pancreatectomy and required 8 to 22 units of long-acting insulin, the series with autotransplants within the omentum, showed a moderate increase in blood sugar (1,1 to 1,1 g/l). However, progressive or acute cachexia with digestive disorders, frequently fistula of the sub-cutaneous graft were observed. In spite of the progressive and attenuated character of the diabetes, the preservation of secreting endocrine pancreatic islets is only temporary as shown by the histological controls.


Applications of high-performance liquid chromatography in the pharmaceutical industry. A survey of the literature has been carried out, covering pharmaceutical applications of high-performance liquid chromatography (HPLC) from 1972-1975. The abstracts are divided into seven groups, viz. alkaloids, antibiotics, nitrogen-containing compounds, steroids, sulphur-containing compounds, formulations, and general analytical techniques. A general resume of the development of HPLC in our own laboratory from 1969 onwards is given together with fourteen chromatograms covering the spectrum of our more novel activities in pharmaceutical analysis.


Effect of melatonin on the basal and stimulated gonadotropin levels in normal men and postmenopausal women. The effect of melatonin on LH and FSH secretion in basal conditions and after stimulation with synthetic LHRH was studied in 3 volunteer normal men and 3 women two to five years after menopause. During the first three days of study, blood samples were obtained at 8 AM. On the second day, an iv injection of 50 mug LHRH was performed; on the third day, 2 ml of 0.9% saline were injected. In both cases, blood samples were obtained 30 and 60 minutes after the injection. On the fourth day, the subjects began melatonin (10 mg daily im for 13 days). Blood samples at 8 AM were obtained after 5, 11, and 13 days of administration of the drug. On days 5 and 11 of the treatment, iv injections of 50 mug LHRH were performed and on day 13 an iv injection of saline 0.9% was given. Blood samples were obtained 30 and 60 minutes after each injection. In each sample LH and FSH levels were determined by radioimmunoassay. Melatonin treatment did not cause any significant change in basal or post-stimulation LH and FSH levels either in men or in post-menopausal women. These results do not support previous findings of an insults do not support previous findings of an inhibitory effect of melatonin on gonadotropin secretion, even with the same dose as the one used in this study. Further studies with higher doses of melatonin are needed to clarify the action of this drug on gonadotropin secretion.


LH and FSH responsiveness to intravenous gonadotropin-releasing hormone (GnRH) in children with hypothalamic or pituitary disorders: lack of effect of replacement therapy with human growth hormone. To evaluate the diagnostic and prognostic usefulness of the GnRH test, gonadotropin responses to iv GnRH (Parke-Davis) were determined in 82 young patients (2,5 mo.-21 yr.) and 6 normal men. After extensive evaluation, 40 patients (31 boys and 9 girls) were considered "endocrinologically normal." Repeat tests were performed in 17 patients at 6-12 mo. intervals. Nine patients with presumed isolated hGH deficiency and 3 patients with multiple pituitary deficiencies were studied before and at the end of 12 mo. of hGH therapy. Serial blood samples were obtained before and after an iv bolus injection of GnRH (2.5 mug/kg, 74 tests, or 10 mug/m2, 36 tests). LH and FSH were determined by radioimmunoassay. Maximum concentration, maximum increment (deltamax), and response area were compared with degree of skeletal maturation to evaluate responses. Clinically, the most useful determination was the deltamax LH. All "normal" children with bone ages greater than 12 yr had LH responses in or slightly above the range of the values of the 6 normal men: deltamax LH, 39 +/- 8 (SE) mIU/ml; range 13-57. Severely blunted or absent responses were seen in 14/15 patients (bone ages 3 mo.-14 yr.) with multiple pituitary deficiencies. Boys with isolated hGH deficiency and bone ages of less than 10 yr had significantly lower responses than "short normal" boys with similar skeletal maturation: deltamax LH, 4.8 +/- 0.9 vs 8 +/- 1.3 mIU/ml, P is less than .05. Although the mean growth velocity of hGH-treated children increased from 3.2 to 8.9 cm/yr, LH and FSH responses were unchanged. These studies indicate that 1) children with idiopathic hypopituitarism (including those with presumed isolated hGH deficiency) have significantly decreased responsiveness to GnRH which does not respond to 6 to 12 months of hGH treatment; and 2) decreased responsiveness to GnRH in patients with bone ages of greater than 12 yr is presumptive evidence of gonadotropin deficiency.


Role of the hypothalamic-pituitary-ovarian axis in puerperal infertility. To gain further insight into the mechanisms of postpartum infertility, plasma gonadotropins, prolactin, estradiol, and progesterone were measured following either iv LHRH or im menopausal gonadotropins (Pergonal) to postpartum women. Plasma estradiol and progesterone rose significantly between 24-72 hours following hMG injection. The response was similar in both nursing and non-nursing women throughout the intermediate and late postpartum study interval. No gonadotropin response was observed following LHRH injection in the early puerperium. Beyond five postpartum weeks, a significantly greater response following LHRH was observed in nursing compared with non-nursing women. A subsequent and significant rise in estradiol correlated with the magnitude of the LHRH-induced increment. Basal prolactin levels in nursing women were significantly greater than in non-nursing women but were unchanged after either hMG or LHRH. We conclude that the postpartum ovary is not refractory to either direct exogenous gonadotropin stimulation or to endogenously released gonadotropins, at least as far as estrogen production is concerned. Furthermore, the difference in the pituitary response to LHRH in nursing and non-nursing women may reflect a difference in the hypothalamic-pituitary axis. Elevated prolactin secretion in response to nursing may be involved in postpartum infertility, but the specific role remaine unclear.


Changing patterns of FSH and LH response to gonadotropin-releasing hormone in the puerperium. Gonadotropin responses to synthetic gonadotropin-releasing hormone (GnRH or LRF), as well as basal concentrations of gonadogropins, estradiol, and prolactin, were determined at weekly intervals during the first eight weeks post partum in seven non-nursing, non-steroid-treated women. The results were compared with those of a group of eleven women studied on the second day of the menstrual cycle. In the postpartum group, baseline concentrations of LH and FSH similar to those seen in the early follicular phase of the menstrual cycle occurred by the third week post partum. In response to an iv bolus of 100 mug GnRH, elevations of serum LH: 1) occurred as early as the second postpartum week; 2) were less than those of women during the early follicular phase until the fourth postpartum week; and 3) were exaggerated (when compared with those ob subjects in the early follicular phase) during the fifth through the eighth week post partum. Similarly, the FSH responses to GnRH in the puerperium were similar to those of the subjects during the early follicular phase by the third week post partum, and were exaggerated during the second month post partum. These studies demonstrate that human pregnancy is followed by a period of relative pituitary refractoriness followed by one of increased responsiveness to GnRH.


Metoclopramide stimulates prolactin secretion in man. Serum prolactin concentrations increased approximately 6-fold after oral or intravenous administration of 10 mg of metoclopramide to adult men. Prolactin remained significantly elevated up to 9 h after oral metoclopramide and for at least 2 h after iv metoclopramide. Bethanechol did not increase serum prolactin, and atropine did not inhibit metoclopramide-induced prolactin elevation, suggesting that the cholinergic-like properties of metoclopramide were not responsible for the observed prolactin responses. Pre-treatment with 500 mg L-dopa inhibited the early metoclopramide-induced prolactin increase, which is consistent with the possibility that metoclopramide acts by inhibiting dopamine-mediated hypothalamic secretion of prolactin inhibitory factor. As a potent stimulus of prolactin release, metoclopramide may be useful in clinical investigation of hypothalamic-pituitary function.


Enzyme-linked immunosorbent assay for streptococcal M protein antibodies. The enzyme-linked immunosorbent assay (ELISA) described by Engvall and Perlmann, which uses antigen-coated tubes and enzyme-labeled anti-immunoglobulin, has been used for the detection of antibodies against streptococcal M protein. The antigen used in the assay was obtained by guanidine extraction of type M-12 streptococcal cell walls followed by hydroxyapatite chromatography. This antigen has the capacity to elicit bactericidal antibodies in rabbits. The results show that the ELISA is specific and highly sensitive for the detection of antibodies in rabbit and human antisera. Preliminary results suggest that, when M-12 antigen is used, the antibodies detected by ELISA are the same antibodies detected in the bactericidal test. The assay has been performed with human and rabbit sera. There was a 96% agreement between bactericidal and ELISA results with rabbit sera and 97.5% agreement with human sera. All bactericidal antibody-positive sera tested thus far yielded positive ELISA results.


Direct measurement of microplates and its application to enzyme-linked immunosorbent assay. A commercially available system for enzyme analysis was adapted for high-speed reading of microplates in which the enzyme-linked immunosorbent assay was performed.


Collagen polymorphism in idiopathic chronic pulmonary fibrosis. Collagens in normal human lung and in idiopathic chronic fibrosis were investigated in terms of their covalent structure and compared for possible alterations in the diseased state. Collagens were solubilized by limited digestion with pepsin under nondenaturing conditions, and after purification they, were fractionated into types I and III. Carboxymethylcellulose and agarose chromatography of both types I and III collagens, and amino acid and carbohydrate analyses of the resulting alpha-chains indicated that the alpha 1 (I), alpha 2, and alpha 1 (III) chains of normal human lung were identical with the human skin alpha-chains in all respects examined except that the normal lung chains contained higher levels of hydroxylysine. Examination of collagens obtained from the diseased lung revealed that the content of hydroxylysine of the alpha 1 (I) and the alpha 1 (III) chains appeared to be diminished as compared to the normal lung chains. The values, expressed as residues per 1,000 residues, are 7.1 and 8.3 for the alpha 1 (I) and the alpha 1 (III) chains, respectively, as compared to 10.0 and 11.1 for the alpha-chains from the normal tissue. The chromatographic properties and amino acid and carbohydrate composition of the alpha-chains from the diseased tissue were otherwise indistinguishable from those of normal lung. In addition, isolation and characterization of the CNBr peptides of alpha 1 (I), alpha 2 and alpha 1 (III) from the diseased lung revealed no significant differences from the CNBr peptides from other human tissues reported previously. Normal and diseased lungs were also digested with CNBr, and the resultant alpha 1 (I) and alpha 1 (III) peptides were separated chromatographically. The relative quantities of these peptides indicate that type III collagen constitutes 33% of the total collagen in normal human lung, with the remainder being type I, whereas in idiopathic chronic pulmonary fibrosis, the relative content of type III collagen is markedly diminished, ranging from 12 to 24% in different patients. These results indicate that an alteration in tissue collagen polymorphism as well as subtle variations in the collagen structure accompany the fibrotic process in the diseased state, and suggest that these alterations may have possible pathogenetic implications.


Hereditary deficiency of the fifth component of complement in man. II. Biological properties of C5-deficient human serum. The first known human kindred with hereditary deficiency of the fifth component of complement (C5) was documented in the accompanying report. This study examines several biological properties of C5-deficient (C5D) human serum, particularly sera obtained from two C5D homozygotes. The proband, who has inactive systemic lupus erythematosus is completely lacking C5, while her healthy half-sister has 1-2% of normal levels. Both sera were severely impaired in their ability to generate chemotactic activity for normal human neutrophils upon incubation with aggregated human gamma-globulin or Escherichia coli endotoxin. This function was fully restored in the sibling's serum, and substantially improved in the proband's serum, by addition of highly purified human C5 to normal serum concentrations. Sera from eight family members who were apparently heterozygous for C5 deficiency gave normal chemotactic scores. The ability of C5D serum to opsonize Saccharomyces cerevisiae (baker's yeast) or Candida albicans for ingestion by normal neutrophils was completely normal. In addition, C5D serum was capable of promoting normal phagocytosis and intracellular killing of Staphylococcus aureus. The proband's serum was incapable of mediating lysis of erythrocytes from a patient with paroxysmal nocturnal hemoglobinuria in both the sucrose hemolysia and acid hemolysis tests, and also lacked bactericidal activity against sensitized or unsensitized Salmonella typhi. The sibling's serum, containing only 1-2% of normal C5, effectively lysed S. typhi, but only at eightfold lower serum dilutions as compared to normals. These findings underscore the critical role of C5 in the generation of chemotactic activity and in cytolytic reactions, as opposed to a nonobligatory or minimal role in opsonization, at least for the organisms under study.


Alterations in T and B lymphocytes in heart transplant patients early and late postoperatively. Alterations in the percent and absolute number of thymus-derived (T) and bursa-equivalent (B) lymphocytes in peripheral blood were followed in 10 patients treated with antithymocyte globulin, prednisone, and azathioprine after cardiac transplantation. During the 1st postoperative wk the percent of T cells dropped below 10% in almost all cases (normal range, 65-91%) with a concomitant rise in the percent of B cells. However, the absolute T- and B-cell counts were both markedly depressed (less than 200 cells/mm3). During the 7-wk postoperative period the percent of T cells rose to 45-60% and the absolute count rose from 100 to 350 cells/mm3 (normal range, 1,092-2,400 cells/mm3). Although the percent of B cells was elevated (35-50%) during this period, the absolute B-cell count remained below the range of normals (268-640 cells/mm3). Follow-up of long-term survivors (3-60 mo postoperative) showed a continued marked T (467 cells/mm3) and B (95 cells/mm3) lymphocytopenia. Chronological relationships between the percent and absolute T-cell count and episodes of graft rejection in individual patients are discussed as possible adjuncts in the prediction of rejection crises.


Defective neutrophil chemotaxis in bone marrow transplant patients. Infection is a frequent cause of death in patients receiving bone marrow transplants. Although lymphocyte dysfunction has been observed in a few such patients, no systematic study of neutrophil function has yet been reported. Neutrophil chemotaxis was evaluated by a 51Cr-radioassay after bone marrow transplantation in 34 patients with acute leukemia or aplastic anemia. The response to a chemotactic stimulus (C5a) was severely depressed (less than 35% of normal) in 18 patients, moderately depressed (35-65% of normal) in an additional 6, and normal in 10 subjects. The mean response in the absence of graft vs. host disease and antithymocyte globulin administration was 73.3+/-9.2% (SE) in contrast to 29.7+/-9.6% (P is less than 0.01) in patients with graft vs. host disease treated with antithymocyte golbulin. Both graft vs. host disease and antithymocyte globulin were implicated since the presence of either factor alone was associated with depressed chemotaxis (31.1+/-4.9% for graft vs. host disease, P is less than 0.01; 17.0+/-7.8% for antithymocyte globulin, P is less than 0.02). When normal neutrophils were incubated with antithymocyte globulin in vitro, their chemotactic response was markedly suppressed in the absence of a cytotoxic effect. Transplant patients with defective chemotaxis experienced significantly more infections than those with normal chemotaxis, and analysis of specific etiologic agents showed that this was predominantly related to bacterial pathogens. Chemotactic inhibitors were detected in the sera of seven patients and elevated IgE levels were found in nine subjects, eight of whom had graft vs. host disease. Generation of chemotactic activity by endotoxin activation of serum was reduced in five patients. The results demonstrate a severe defect in neutrophil chemotaxis in some bone marrow transplant patients and suggest that neutrophil dysfunction may predispose to infection in such patients.


Disturbed islet-cell function related to endogenous gastrin release. Studies on insulin secretion and glucose tolerance in pernicious anemia. The insulin and gastrin response to oral glucose, intravenous glucose, or a protein-rich meal were measured in 44 nondiabetic patients with pernicious anemia (PA) and in 44 control subjects. 36 of the PA-patients had hypergastrinemia, while serum gastrin concentrations in the remaining eight patients were below normal. Three hypergastrinemic PA-patients were in addition studied during an oral glucose loading with synchronous intravenous infusion of gastrin-17. During both oral and intravenous glucose tests blood glucose concentrations were similar in patients and in controls. After ingestion of protein blood glucose concentrations in PA-patients with hypergastrinemia were above those of the controls (P less than 0.05). Parenteral infusion of gastrin-17 during oral glucose loading also increased blood glucose concentrations above the levels observed after glucose alone. In PA-patients with hypergastrinemia the insulin response was augmented in all tests. In patients with hypogastrinemia serum insulin concentrations were lower than normal in the fasting state and during stimulation with glucose intravenously (P less than 0.01). In hypergastrinemic patients serum gastrin concentrations decreased after oral as well as intravenous glucose administration. The decrease was larger during the oral test. In hypogastrinemia oral glucose induced, as in controls, a small initial rise followed by a slow fall in serum gastrin concentrations. No variations were seen in these patients during the intravenous glucose infusion. Gel filtration of serum from hypergastrinemic patients disclosed a decrease in the concentrations of all four main components of gastrin during the glucose loadings. Taken together with earlier studies on the effect of exogenous gastrin the results suggest that endogenous hypergastrinemia induces hyperglycemia and potentiates insulin secretion. In contrast hypogastrinemia is associated with hypoinsulinism.


Clinicopathological spectrum of late postpartum renal failure; two contrasting cases. The clinical and renal biopsy findings from two patients in whom renal functional abnormalities developed in the late postpartum period are described. Both biopsies showed fibrin deposition in the renal vasculature, in one case marked and in the other mild. The patient with the more severely damaged kidney subsequently died, and the other is alive but with evidence of slowly progressing renal damage. The clinicopathological spectrum and pathogenesis of late postpartum renal failure are discussed.


Re-evaluation of heat precipitation method for plasma fibrinogen estimation: effect of abnormal proteins and plasma viscosity. The heat precipitation method (Aberdeen method) was compared with the Ratnoff and Menzie method of fibrinogen assay in 320 donors, including normals and patients suffering from malignant melanoma, renal failure, hypertension, multiple myeloma, etc. Excellent correlation (r=0-8287, p less than 0-000 000 1) was found between these two methods. However, on some occasions individual low results were obtained by the Aberdeen method in the presence of cryoglobulins or excessively high plasma viscosity. The latter effect was tested also by additions of albumin, glucose, and dextrans.


Significance of subnormal red-cell folate in thalassaemia. Subnormal red-cell folate values were encountered in 20 of 63 thalassaemic subjects in a population with a low incidence of megaloblastosis. The abnormality was not associated with haematological changes of megaloblastosis and could not be explained on the basis of incomplete liberation of intra-erythrocytic folates, serum conjugase deficiency of vitamin B12 deficiency. Evidence is presented to indicate that it is unlikely to represent a state of subclinical folate deficiency. The exact cause of the abnormality is unknown.


Weber-Christian panniculitis and auto-immune disease: a case report. A case is described of Weber-Christian panniculitis accompanied by a gammaglobulin disturbance which preceded by five years the diagnosis of an autoimmune hepatitis and pancytopenia. Also associated was the onset of diabetes mellitus, found at necropsy to be related to pancreatic islet amyloid deposition. This case reinforces the view that Weber-Christian panniculitis may be an adipose response to a variety of immunological stimuli.


A comparison of two commerical methods for the identification of the Enterobacteriaceae--API 20E and the Enterotube--with conventional methods. The API 20E, the Enterotube, and routine methods of Cowan and Steel were used in parallel to identify 245 members of the Enterobacteriaceae. The API 20E and conventional methods gave the same identification in all but 2 (0-8%) of the 245 organisms tested. The Enterotube correctly identified 85% of these organisms on the first testing. On re-testing those organisms incorrectly identified the Enterotube results agreed with the conventional ones in a further 20 (8%). There was no change in the identification obtained by the API or conventional methods. Further conventional sugar tests were necessary before final identification was available by the API system in 7 (3%) against 106 (47%) of the 226 organisms correctly identified by the Enterotube. The Enterotube relied on serological testing alone to distinguish between alternatives in 17 (7%) isolates. Other advantages and disadvantages of these systems are discussed.


Activity and interaction of trimethoprim and sulphamethoxazole against Escherichia coli. The activity of trimethoprim (TMP) and sulphamethoxazole (SMX), alone and in combination, against a sensitive strain of Escherichia coli was investigated in turbidimetric systems. In a static system in which the conditions of exposure of bacteria to drug resembled those of conventional minimum inhibitory concentration (MIC) titrations, both TMP and SMX exhibited antibacterial activity at concentrations well below the conventionally determined MIC, but regrowth occured at these concentrations during the overnight incubation period due to the emergence of adaptively resistant bacteria. Tests of combined drug action in the static turbidimetric system revealed even more synergic interaction than was apparent in conventional MIC tests. It is suggested that an important component of overall synergic interaction is the mutual suppression of adaptive "resistance" to the other agent. Studies in an in vitro model which simulates the hydrokinetic features of the urinary bladder showed that concentrations of TMP and SMX below the conventionally determined MIC inhibited the growth even of extremely dense bacterial populations so long as the concentration of drug was maintained. The response of cultures exposed to combinations of TMP and SMX in this system was so dominated by the effect of TMP that no synergic interaction with SMX was noted at concentrations of the drugs which are achievable in urine.


Hepatitis B surface antigen (HBsAg) in the liver of patients with hepatitis; a comparison with serological detection. Chronic hepatitis was diagnosed on liver biopsy of 76 patients; 52 (68%)had HBsAg. Of the 52 patients with HBsAg, 23% had HBsAg shown by immunofluorescence on the liver, while it could not be detected with radioimmunoassay on the serum; 77% had HBsAg detectable in liver and in serum, and none had HBsAg in serum only. HBsAg was detected more frequently in chronic aggressive hepatitis and active cirrhosis than in chronic persistent hepatitis and cirrhosis with little activity. No correlation was found in the different forms of chronic hepatitis between the HBsAg status on the one hand, and levels of transaminases, gammaglobulins, and auto-antibodies on the other. Acute hepatitis was diagnosed on liver biopsy of 24 patients; 50% had HBsAg. Liver tissue positivity was very low in the fully developed stage compared to serum positivity. In 146 patients with other liver ailments, both liver and serum were negative for HBsAg.


Distribution patterns of hepatitis B surface antigen (HBsAg) in the liver of hepatitis patients. One hundred liver biopsies from 100 hepatitis patients were examined by the indirect immunofluorescent technique for the detection of HBsAg. Of the 60 positive specimens 52 were diagnosed as various types of chronic hepatitis and 8 were acute hepatitis. Four main distribution patterns of HBsAg were obtained: full cytoplasmic fluorescence with diffuse lobular distribution; cytoplasmic fluorescence with spotty distribution; peripheral fluorescence in the cell membrane and/or cell peripheries; and focal cytoplasmic positivity. There was an inverse relationship between the number of positive hepatocytes and the extent of liver cell necrosis. The distribution patterns of HBsAg were distinctive in each type of chronic hepatitis and in acute hepatitis. Homogeneous full cytoplasmic fluorescence, distributed diffusely in the whole liver lobule, was observed in chronic persistent hepatitis and in cirrhosis with little activity whereas peripheral liver cell membrane and/or peripheral cytoplasmic fluorescence associated with cytoplasmic positivity in a smaller number of hepatocytes was a characteristic finding in chronic aggressive hepatitis, active cirrhosis, and acute hepatitis with possible transition to chronicity. Focal cytoplasmic fluorescence was observed in acute hepatitis and a group of biopsies in chronic hepatitis in which HBsAg was detected in the liver but no antigen was detectable in the serum. The results show that the different patterns of distribution of HBsAg in the liver biopsy are helpful for the histological diagnosis of different types of HBAg positive viral hepatitis and are consistent with the hypothesis of the role of specific immune response in the pathogenesis of type B viral hepatitis.


Comparison of laboratory methods in the diagnosis of urinary tract infection. Three methods of semiquantitative culture and two techniques of microscopy were compared with a surface viable count for the detection of significant bacteriuria in one thousand midstream specimens of urine. The results obtained with the blotting-paper-strip method on MacConkey agar and with Uricult dip-slides correlated well with the results of the surface viable count, and both methods were suitable for routine use. The blotting-paper-strip method was preferred for laboratory use because of expense but dip-slides are useful for general practice and outpatient clinics. Semi-quantitative culture by Microstix dip-strips gave less accurate results, and the nitrite test area detected only a small proportion of infected specimens. The microscopic examination of a Gram-stained film of the centrifuged deposit of urine specimens yielded more useful information and was more reproducible than examination of a wet film of the untreated urine.


A new serotyping method for Klebsiella species: development of the technique. A new serotyping method for Klebsiella species using indirect immunofluorescence is described. Nonspecific fluorescence has been minimized by carrying out the capsular antigen-antibody reaction at pH 9.0. Commercial antisera have been tested with the 72 antigenic types of Klebsiella, and appropriate dilutions of each pool and specific antisera have been proposed for use in routine typing. Dilutions were chosen to allow strong fluorescence with each type and its specific antiserum and minimal fluorescence with cross reacting antisera. Where the pool antisera gave a weak reaction for one or more of the component types, it is recommended that the specific antisera for these types be added to the pool dilution. The few remaining cross reactions, with the pool and specific antisera in test dilution, are listed in a table. The unique cross reacting patterns of particular types have been found to be useful in identification. Typing Klebsiella by the fluorescent antibody technique is easy to perform and interpret; the results are reproducible, and it is less expensive than the existing capsular swelling method as it is more sensitive and requires less concentrated antisera. This new method of typing should facilitate detailed epidemiological studies of the mode of transmission of Klebsiella species in hospitals and thus allow more effective infection control measures to be instituted.


A new serotyping method for Klebsiella species: evaluation of the technique. A new indirect fluorescent typing method for Klebsiella species is compared with an established method, capsular swelling. The fluorescent antibody (FA) technique was tested with standards and unknowns, and the results were checked by capsular swelling. Several unknowns were sent away for confirmation of typing, by capsular swelling. The FA method was also tried by a technician in the routine department for blind identification of standards. Fluorescence typing gives close correlation with the established capsular swelling technique but has greater sensitivity; allows more econimical use of expensive antisera; possesses greater objectivity as it requires less operator skill in the reading of results; resolves most of the cross reactions observed with capsular swelling; and has a higher per cent success rate in identification.


Infection with Streptococcus agalactiae in a London hospital. Streptococcus agalactiae is presently the commonest beta-haemolytic streptococcus isolated from clinical material in this hospital. Between October 1974 and March 1975, 81 patients with such infections were seen. Seventeen had urinary tract infections, six had septicaemia, and one neonate had meningitis. Three of those with septicaemia were neonates and two died. The organism was also found to be a cause of pyogenic skin conditions in five patients. Isolates from throat swabs in 12 patients, from sputum in four, and from the female genital tract in 18 were considered part of the normal flora. Human strains of Str. agalactiae were found to be biochemically different from animal strains.


Evaluation of automated large-scale screening tests for syphilis. Two methods of performing serological screening tests for syphilis are compared. One consisted of the Venereal Diseases Reference Laboratory (VDRL) slide test, the cardiolipin Wassermann reaction (CWR), and the Reiter protein complement fixation test (RPCFT) performed manually; the other was a fully automated system using two Technicon AutoAnalyzers (AAII), one for the automated reagin test (ART) and the other for automated complement fixation tests. The absorbed fluorescent treponemal antibody test (FTA-ABS) was used as a final arbiter in all cases found to be seropositive by either method. A pooled antigen consisting of a mixture of cardiolipin and Reiter protein was used for the automated complement fixation test, thus increasing the scope and capacity of the system. The AutoAnalyzer was shown to be capable of performing 400 cardiolipin and Reiter complement fixation tests and 700 automated reagin tests in an 8-hour day. Modification of the complement fixation test method to take advantage of the highly sensitive colorimeter resulted in a significant increase in sensitivity and a corresponding saving in reagents. Of the 7843 sera tested, 258 gave a positive result in one or more of the screening tests. The automated test detected many more Reiter positive sera (127) than the manual test (83). Conversely, fewer CWR positive sera were detected by the automated test (60) than by the manual test (82). There was little difference between the number of positive sera detected by the ART (73) and the VDRL slide test (71). In 19 instances the automated tests detected positive sera which registered as completely negative in the manual tests, and four seropositive cases which the automated tests had failed to detect were detected by the manual tests, and four seropositive cases which the automated tests had failed to detect were detected by the manual tests. It was concluded that a combination of the ART and automated Reiter protein complement fixation test (ARPCFT) would be ideal for use in a large-scale screening programme for the detection of syphilis.


Classification of smooth muscle autoantibodies detected by immunofluorescence. Three hundred and twelve sera containing antibodies to smooth muscle (SMA) wer analysed for the immunofluorescence patterns they produced in various tissues. A classification is described based on the three main appearances in rat kidney. Some sera, mainly of low titre, reacted only with vessel walls (SMA-V), some stained vessels and renal glomeruli (SMA-G) and high titre sera, mainly from patients with chronic active hepatitis stained vessels, glomeruli, and intracellular fibrils in renal tubules (SMA-T). Peripheral staining in hepatocytes or thyroid cells was not a regular feature. 41/43 polyclonal SMA-T and -G sera were absorbed out completely by actin, and this also removed the pericullular staining in liver and thyroid when present. High titre SMA-V antibodies could not be absorbed by actin, and the antigen remains to be identified.


Rapid diagnosis of anaerobic infections by gas-liquid chromatography of clinical material. Gas-liquid chromatographic analysis of samples of pus provides a rapid and reliable means for the presumptive differentiation of anaerobic from aerobic infections.


Instability and linkage of silver resistance, lactose fermentation, and colony structure in Enterobacter cloacae from burn wounds. A strain of Enterobacter cloacae from a burn wound has displayed unstable resistance to silver nitrate and a linkage of this characteristic to that for rapid lactose fermentation and mucoidcolony structure. Attempts to demonstrate transferability of these unstable determinants gave negative results.


Intraoral cancellous bone autografts in the treatment of infrabony pockets. One hundred infrabony pockets with one- and two-wall bony defects were treated for reattachment. Mucoperiosteal flaps were raised, the tooth surfaces were scaled and planed, and the defects curetted. Flaps were sutured tightly and the sites protected with a surgical dressing. Antibiotic coverage was used in each case. Pre- and postoperative measurements were taken by the same clinician from the cemento-enamel junction to the base of the pocket. In 56 defects cancellous bone from the same patient was placed in the defect, and in 44 defects the treatment consisted only of open curettage without bone grafts. The results showed a trend towards more favourable clinical results using bone grafts; especially in the two-wall bony defects.


Ultrasonic scaling of maxillary teeth causing tinnitus and temporary hearing shifts. The possible effects of ultrasonic scaling of all maxillary teeth for a total of 5 min on the inner ears of 20 healthy young adults, 22-36 years of age, were investigated by means of pure tone audiometry. Audiograms were obtained just before and immediatley after scaling. Temporary shifts in hearing threshold (TTS) of 10-20 dB, mostly at 7--8 kHz, ersisting for up to 30 min, were recorded in eight of the individuals. Three patients reported high-pitched tinnitus lasting for 20-30 min after ultrasonic scaling. When audiograms were obtained 3--5 weeks later in the same 20 individuals before and immediatley after they had kept their mouths open for 5 min, but with no scaling performed, TTS was recorded in three of the individuals. TTS after opening only was less pronounced than TTS after ultrasonic scaling, both with regard to severity and duration.


Health maintenance organizations: are they for the inner cities? Health Maintenance Organizations present some major limitations as a means to address the health care needs in our nation's inner cities. The HMO as it has been affected by HMO legislation is discussed and an identification made of those areas that may adversely affect the delivery of health services to inner-city residents where costs may be greatest for those who can least afford it.


Quality of health care for the disadvantaged. Literature review points out that: (a) differentials in health status between the disadvantaged and the nondisadvantaged persist, often to a large degree; (b) differentials in the overall amount of care received are less striking now than heretofore, but standardization by level of need demonstrates measurable discrepancies in health services provided to the disadvantaged compared with the nondisadvantaged; (c) the quality of health care for the disadvantaged is not strikingly poorer than care for the nondisadvantaged, but, in view of demonstrable shortcomings in the quality of health care in general, this is not viewed as a positive statement;  and (d) attempts to improve quality of care for the disadvantaged have not had the hoped-for impact. Four new avenues are suggested for possible further research; increased patient responsibility, increased consumer knowledge, financial accountability, and quality assurance activities. Because of the likelihood of only marginal changes in health status, rigorous evaluation of any experimental program is emphasized. During the last decade, many attempts have been made by private and governmental bodies to improve the health of the American people. In general, these efforts have focused on improving the health of members of disadvantaged groups and have included such diverse activities as building OEO health centers, developing maternal and infant care programs, and financing care for the elderly. During the last few years, a different movement, concerned with assuring high quality care for all people, has produced efforts such as quality assurance activities in health maintenance organizations, the Professional Standards Review Organization program, and the medical care evaluation program of the Joint Commission on the Accreditation of Hospitals. Consideration of these two issues, i.e., improving the health of disadvantaged groups and improving the quality of care for all people, has led to two policy-relevant questions: "Can the health of disadvantaged groups be substantially improved by assuring that a high level quality of care is delivered to them?" and "Can the quality of care delivered to disadvantaged groups be improved?" The purpose of this paper is to review some available data pertinent to both these issues and to suggest some ideas for future research.


Ultrastructural findings in bullous pemphigoid. In four bullous pemphigoid patients electron microscopic and direct immunofluorescence examinations were carried out on sections of the border of a blister, apparently normal skin surrounding the blister, apparently normal skin distant from the blister, and apparently normal skin subjected to the stimulus of a xenon lamp. The bullous pemphigoid blister was formed by cleavage of the basal cells away from the basal lamina which remained intact. The intercellular spaces were widened and the half-desmosomes reduced in number in the skin surrounding the blister. Xenon lamp irradiation produced a pattern similar to that of the skin surrounding the blister, but without immunoglobulins and complement fixed in vivo.


Male pattern alopecia a histopathologic and histochemical study. Three hundred and forty-seven tissue specimens were studied from 23 patients with male pattern alopecia. Characteristic features of pattern alopecia included: the presence of miniature or vellus follicles; a marked enlargement of the sebaceous glands and arrectores pilorum muscles; the presence of connective tissue streamers beneath the vellus follicles; and the thinning of the dermis. A mild perivascular infiltrate of mononuclear cells and mild capillary dilatation was sometimes seen. An increased number of mast cells was often a prominent feature. Histochemical procedures were performed for glycogen, acid mucosaccharides, inorganic substances, and enzymes including alkaline phosphatase, acid phosphatase, beta glucuronidase, cholinesterase, aminopeptidase, oxidases and dehydrogenases. Histochemical studies did not reveal any significantly abnormal enzyme changes other than the altered vascular and nerve supply to the the miniature follicles.


Effect of coliform challenge at milking time on new udder infections. This project was designed to study rates of infection in udders of cows exposed to an Escherichia coli broth culture at milking time. Forty Holstein cows of varied stages of lactation were divided randomly into three treatment and one control group of ten cows each. The treatment groups were exposed for 3 wk to an Escherichia coli broth of 10(9) colony forming units per ml at milking time by either 1) dipping teat ends in broth before milking, 2) spraying the udder and leaving it dripping wet during milking, or 3) dipping teat ends in broth after milking. Eleven of 30 treated cows became infected in one or more quarters; all control cows remained uninfected. The infection rate of the three Escherichia coli-treated groups was higher than the controls. However, there was no difference among treated groups. Exposure to the broth culture of Escherichia coli increased the infection rate, but the time at which the udder was exposed to the organisms was unimportant. All infections were of the same type with the same O and H group antigens as the Escherichia coli broth.


Wedge resection of the lower lip.

Wedge resection of the lower lip. Wedge resection of tumors on the lower lip can be done as an office procedure. The operative technique with special emphasis on realigning the vermilion is described.


Proper biopsy technique for immunofluorescence tests on skin. Successful use of direct immunofluorescence tests on skin depends on selection of the appropriate biopsy site, careful preparation of the specimen, and prompt transportation to a reliable laboratory. Recommendations for selection of biopsy sites in various dermatoses and methods available for handling specimens are discussed.


Magnification--an interesting optical property of dentin. Disks of dentin taken from extracted human teeth demonstrated the optical property of the magnification/reduction effect. When images were transmitted from a pulpal-to-occlusal direction, the images were magnified. When the disks were inverted, the images were reduced in size. The images were slightly distorted but clear. When these disks were decalcified or dehydrated, they still possessed the same magnification/reduction effect property. The clear image transmission is attributed to the passage of light along the dentinal tubules; the magnification is the result of their divergence from the pulpal to the occlusal direction. The degree of tubule divergence was determined to correspond closely with the proportion of magnification and of reduction. In calcified sections, an individual tubule would act as a light-pipe, conducting light as it reflects internally off the dentin walls. Decalcified disks may demonstrate a fiberoptic effect. The magnification of an image is the result of the divergence of hundreds of thousands of minute light conductors.


An improved mattress suture.

An improved mattress suture. An improved mattress suture that is completely buried and leaves no noticeable cross marks on either side of the incision is described.


Xylitol-induced increase of lactoperoxidase activity. Whole saliva samples of volunteers who ate a strict diet for two years with regard to the type of sweeteners used (sucrose, fructose, and xylitol) showed considerable differences in the lactoperoxidase activity. The consumption of a xylitol diet increased the activity of this enzyme fourfold to tenfold when compared to the other two test groups. Lactoperoxidase belongs to the natural defense mechanisms of the oral cavity. However, the consumption of a xylitol diet also leads to a strong reduction in the incidence of dental caries. It is suggested that the xylitol-induced elevation of the salivary lactoperoxidase activity and the cariostatic properties of xylitol are partly interrelated phenomena.


Endodontics 1776-1976: a bicentennial history against the background of general dentistry. During 200 years of progress in endodontics, various sciences have contributed to our understanding of the physiology and pathology of the dental pulp. Early treatment included cauterization of the pulp, the use of poultices or leeches, and tooth transplantation or replantation. Various methods of pulp devitalization are reviewed. Near the turn of the century, the discovery of X rays made diagnosis more accurate and the discovery of local anesthetics eliminated pain during endodontic treatment. Although the focal infection theory slowed the acceptance of endodontic treatment in this century, the biomechanical concept of treatment and research have recently opened new avenues for treatment and have initiated improvements in medicaments and filling materials.


Significance of tartrazine sensitivity in chronic urticaria of unknown etiology. Of 38 patients with chronic urticaria of unknown etiology who were evaluated for food and drug additive sensitivity, 53% (20/38) had urticaria for 1 yr or more. Total eosinophil counts were not elevated in most patients, and the frequency of atopy was found to be similar to that in a general population. Of these 38 patients, 10 (26%) had a personal history of aspirin intolerance, but elimination of aspirin did not relieve the urticaria. In a double-blind crossover challenge with 0.22 mg of tartrazine and a control, tartrazine sensitivity was found in 8% (3/38) of patients with chronic urticaria and 20% (2/10) of patients with aspirin intolerance.


Food preparation in colonial America. A Bicentennial study. Both regional and national influences have pervaded America's culinary arts from colonial times until the present. In the South, for instance, indigenous foods, such as sweet potatoes--as well as an abundance of fruits and fowl--were commonly served. In the North, maple sirup was a New England product, as was codfish. Throughout the colonies, corn was easily grown and became a staple. Immigrants from the Old World brought their recipes to  meld or adapt to conditions they met here. Recounted also is the unfolding of an American cuisine, especially in the southern colonies as it evolved from European food preparation practices. Cooking was done in great fireplaces, with equipment designed to fit. Meat was generally boiled or stewed in pots hung in the fireplace, although it might be slow-roasted on a hand-turned spit. Hot breads, the hallmark of southern cooking, date from colonial days. In the Noth, the Dutch farmer's wife developed real skill in using flour from home-grown wheat and rye, creating pancakes, waffles, doughnuts, crullers, and so on. After the first hard winter, food in New England became more plentiful. Boston brown bread was made from corn, wheat, or rye and probably sweetened with maple sirup. Imports of coffee, tea, and spices from the Orient and fruit from the tropics were later added to the cuisine. Colonial Americans understood well the art of food preparation and appreciated the taste of well prepared, well seasoned dishes.


Isolation of salmonellas and Shigella sonnei from a laboratory bench. An area of the laboratory bench on which slide agglutinations were performed in the diagnosis of salmonella and shigella infection was examined for these organisms. Impression plates and broth-moistened swabs were used for sampling. Both techniques gave satisfactory results, but the contact plates provided positive results a day earlier than the swabs. Suitable precautions to minimize contamination of the bench surface are discussed.


The laboratory evaluation of iodophor disinfectants with yeast suspensions. The report investigates the variation in results noticed when testing iodophor disinfectants in the presence of a 5% (w/v) yeast suspension. It was found that these variations were not related to individual iodophor formulations but bore a direct relationship with the storage time of the prepared yeast suspension.


Bacterial dispersion in relation to operating room clothing. The effect of operating clothing on the dispersal of bacterial particles from the wearers was studied in a dispersal chamber. A comparison was made of six gowns as well as four types of trousers. The gowns were of three basic types, namely a conventional cotton type, disposable types made of non-woven fabric and those of the total-body exhaust system (Charnley type). The dispersal chamber could simulate conditions as expected both in down-flow unidirectional ultra-clean systems and in a conventional turbulent plenum-ventilated system. It was found that the disposable gowns would reduce the dispersal rate by about 30% in the simulated conventionally ventilated system and about 65% in the laminar flow system. The total-body exhaust system (Charnley) would reduce the count by 10-fold in the conventional ventilated system and by 66-fold in the laminar-flow system. The poor performance of the gowns in conventionally ventilated systems was caused by the dispersal of bacterial particles from underneath the gown (about 80%). This was not reduced by the disposable gown and only partially by the Charnley type. This small drop would be further decreased in a conventionally ventilated operating-room as only scrubbed staff would wear the gown. In order to overcome this poor performance in conventionally ventilated operating-rooms impervious trousers would be required. Four types were studied and it was demonstrated that those made either from Ventile or non-woven fabric would reduce the bacterial dispersion fourfold. As these tests had been carried out in an artificial environment checks were carried out in the unidirectional-flow operating-room during total-hip arthroplasty. This was done by comparing conventional cotton gowns with non-woven gowns and total-body exhaust gowns. The results showed good correlation between the operating room and the chamber with the non-woven fabric gown but the total-body exhaust system did not perform as well in the operating room (12-fold compared to 66-fold) the difference being possibly due to the contribution from the patient. However, as this comparison was that which would be most open to influence from other variables confidence could be placed on the chamber test results. Values were also obtained for the total number of bacterial particles dispersed by persons during a standard exercise wearing different clothing. This count was dependent on the clothing worn but a median count of between 1000 and 1500 bacterial particles/min. would be expected when conventional clothing was worn, with a range of between 300 and 19,000. This count could be reduced to about 100/min. if a total-body exhaust suit was worn (range 30-400).


The distribution of serotypes of Escherichia coli in cow-pats and other animal material compared with serotypes of E. coli isolated from human sources. The serotypes of 13,139 strains of Escherichia coli isolated from humans were compared with the serotypes of 1076 strains isolated from animals. 689 of these strains were isolated from fresh cow-pats on 22 sites in England and Wales. 708 different O/H combinations were found. Of these, 520 were found in human strains only, 130 from animal strains only and 58 O/H serotypes from humans and animals. Approximately half of the animal strains could not be typed with the full set of sera used.


Inhibition of antigen-induced B lymphocyte activation in vitro by cytochalasin B. Cytochalasin B (CB), a fungal metabolite which disrupts microfilaments. and will inhibit capping and other events requiring membrane movement, suppressed the production of antibody-forming cells (AFC) in both mouse whole spleen cultures immunized with sheep erythrocytes (SRBC) and in mouse B lymphocyte cultures immunized with the thymic independent antigen DNP-Ficoll (DF). CB at a concentration of 1 mug/ml inhibited the AFC response by more than 90% in spleen cell cultures immunized with SRBC. This inhibition was completely reversible by removal of CB up to 24 hr after the start of culture. Spleen cells cultured in the presence of CB for the first 48 to 72 hr had a decreased AFC response similar to that of cultures in which SRBC had been withheld for thocyte the same period of time. Incubating whole spleen cell or B lymphocyte cultures immunized with DF for as short as 6 hr decreased the AFC response more than 60%. Antibody secretion, cell viability, and antigenicity of the SRBC and DF were not affected by CB. The results of these experiments favor the concept that movement of surface receptors is necessary in activating lymphocytes to differentiate into AFC. The differential response to CB observed in SRBC and DF stimulated cultures makes the technique employed a useful tool to study membrane events occuring between antigen interaction with surface receptor and the initiation of differentiative events.


Humoral immunity in experimental syphilis. I. The demonstration of resistance conferred by passive immunization. Evidence of a role for human factor(s) in immunity to experimental syphilis has been provided by the demonstration that passive immunization of rabbits by daily i.v. injections of immune serum significantly delays the appearance and markedly diminishes the severity and duration of lesions which develop after challenge with Treponema pallidum. Five rabbits were injected daily over 37 days with 3 ml/kg body weight of pooled immune rabbit serum injection with 1.1 X 10(3) T. pallidum, Nichols strain, at each of four sites. The animals developed atypical lesions of short duration after an average delay in onset of 28 days short duration after an average delay in onset of 28 days beyond the development of typical lesions in control animals similarly injected with nonimmune serum or saline. The failure of passive immunization to provide complete protection was evident not only in the development of the atypical lesions, but also in the demonstration of disseminated infection in the tissues of three of the four surviving animals 7 months after challenge. The possibility that incomplete protection may have been due to 1) insufficient immune serum levels, 2) intracellular location of T. pallidum, and/or 3) cell-mediated mechanisms is discussed.


Humoral immunity in experimental syphilis. II. The relationship of neutralizing factors in immune serum to acquired resistance. Evidence for a humoral mechanism in immunity to experimental syphilis was provided by the demonstration of immune rabbit serum factor(s) capable of inactivating virulent Treponema pallidum, Nichols strain, in an in vitro-in vivo neutralization test. After intratesticular infection, rabbits were bled periodically and their resistance to reinfection was determined by challenge with T. pallidum. The results of challenge showed that resistance to reinfection begins to develop by 11 days after infection, becomes complete by 3 months, and persists for at least 2 years. In the neutralization test, a mixture of treponemal suspension and serum from the infected animals was incubated anaerobically at 34 degrees C and the virulence of the treponemes was determined by intradermal inoculation into normal rabbits. Complete inactivation of treponemes by immune serum required heat-stable and heat-labile (56 degrees C, 30 min) serum components and 16 hr of incubation, and was accelerated by pre-incubation of the treponemes for 4 hr with nonimmune serum but not by 100 mug/ml of added lysozyme. Serum-neutralizing activity, first demonstrable 1 month postinfection, was quantitated by a neutralizing endpoint (NEP). A relatively close quantitative correlation was shown between the development of resistance to symptomatic reinfection and the appearance and persistence of both TPI antibody and neutralizing serum factor(s). The nature of the serum factor(s), the mechanism of treponemal inactivation, and the application of the test in assessing the immune status are discussed.


Eosinophil chemotactic factor (ECF). I. Release from polymorphonuclear leukocytes by the calcium ionophore A23187. Eosinophil chemotactic factor (ECF), previously thought to be primarily associated with human basophils and mast cells, could be released from human neutrophils (PMN) and eosinophils but not lymphocytes by the calcium ionophore A23187. Release of ECF from PMN was time and dose-dependent. Like antigen-induced, basophil-derived ECF, PMN-derived ECF had a high selectivity for eosinophils as determined by differential counts of migrating cells and by deactivation studies. Chromatographic analysis of PMN-derived ECF on Sephadex G-25 showed an elution pattern very similar to that of basophil-derived ECF. With rat peritoneal cells, it was possible to show that mast cells as well as mast cell-depleted cell preparations could be induced to release an ECF that appears to be similar or identical to human ECF. These findings suggest that ECF may play a role in inflammatory processes involving cells other than basophils and mast cells.


Detection of antibodies to bacterial cell wall peptidoglycan in human sera. A radioimmunoassay has been developed for the measurement of antibodies to peptidoglycan in human sera including patients with rheumatic fever and juvenile rheumatoid arthritis. The assay is based on the percentage of binding of the hapten 125I-L-Ala-gamma-d-Glu-L-Lys-D-Ala-D-Ala, the major peptide determinant of peptidoglycan. Because of differences in the avidity of the antibodies in different sera, the amount of antibody was expressed as pentapeptide hapten-binding capacity (pentapeptide-HBC in ng/ml of serum). Fourteen out of 105 normal blood donors had a pentapeptide-HBC value greater than or equal to 75 ng/ml serum. Values in healthy children 5 to 18 years of age were less than or equal to 50 ng/ml. Sixty-eight percent of the individuals with rheumatic fever had values greater than or equal to 75 ng/ml, an indication that streptococcal infections can stimulate an immune response to peptidoglycan. Thirty-five percent of the patients with juvenile rheumatoid arthritis had values greater than or equal to 75 ng/ml. Such a finding points to a possible association between bacterial infections and juvenile rheumatoid arthritis.


The xenogeneic effect. II. Requirement for unactivated murine T cells during restoration of immune responsiveness with xenogeneic reconstitution factor. Studies were conducted on the mechanism of action of a soluble mediator which was generated in human mixed lymphocyte cultures and assayed for helper activity in T cell-deficient murine spleen cell cultures. The mediator, termed xenogeneic reconstitution factor or XFR, restored the anti-sheep erythrocyte plaque-forming cell response of spleen cells from thymectomized, lethally irradiated, and syngeneic bone marrow transplated (TxB) mice. It was found that in order to obtain a maximal antibody response, the XRF had to be present sometime during the first 24 to 40 hr of the induction process. XRF was also required during the last 24 hr of the incubation period. A striking synergistic effect was obtained by combined exposure of the T cell-deficient spleen cell cultures to XRF during the first and last days of culture. These data suggested a bi-modal mechanism of action of XRF, and raised the possibility that in addition to providing a signal to the B cells, XRF-mediated activation of the residual TxB splenic T cells was crucial to the successful restoration of the antibody response. Treatment of the nonadherent splenocytes from the TxB mice with anti-T cell serum and guinea pig complement completely abrogated the antibody response of these cells in the presence of adherent spleen cells, sheep erythrocytes, and the helper factor XRF. The antibody response of this combination of cells, antigen, and XRF was reconstituted by a population of unimmunized murine T cells which, in the absence of XRF, were totally unable to restore responsiveness. These results suggest that B cell activation to the formation of antibody involves the mandatory co-participation of two functionally distinct helper activities.


Visualization of type I and II collagens in tissue sections by immunohistologic techniques. Rabbit and rat antibodies were prepared against Type I and II collagens derived from bovine skin and articular cartilage, respectively. As judged by passive hemagglutination and radioimmune assays, these antibodies could be rendered generally specific for the type of collagen used for immunization by immunoadsorption. Thus, antibodies to Type II collagen did not cross-react with Type I and III collagens from skin. However, antibodies to Type I collagen still showed some cross-reaction with Type III collagen. Antibodies to Type I procollagen showed a negligible degree of cross-reaction with Type III procollagen. These purified antibodies reacted strongly with bovine and human tissue collagen as demonstrated by indirect immunoflourescence. Antibodies to Type I collagen stained dermal tissue, perichondral tissue, kidney stroma, aortic tissue, and annulus fibrosus. Antibodies to Type II collagen stained mainly the hyaline matrix of rib cartilage and nucleus pulposus. The staining patterns with anti-Type I procollagen were similar but not identical to that found with antibodies to Type I collagen. Neither of these antibodies reacted with kidney glomerular basement membrane. These antibody reagents are recommended as a sensitive and rapid screening tool for studying tissue distribution of collagen under normal and pathologic conditions.


The course of Schistosoma mansoni infection in thymectomized rats. Inbred rats were thymectomized, irradiated, and reconstituted with T cell-free bone marrow cells. Thymectomized-reconstituted (B rats) and control rats were infected with Schistosoma mansoni cercariae and the number of worms recovered was determined at various times after infection. The extent of immunosuppression was assessed by two criteria: 1) response to an injection of sheep erythrocytes (plaque assay, hemagglutination, hemolysis); 2) response to schistosome antigens (passive hemagglutination). Humoral responses to worm antigens were completely suppressed in almost all instances and anti-sheep erythrocyte responses showed a more variable but always very definite depression in B rats. The number of worms in B rats was about 4 times higher than in control animals at 5 weeks and about 3 times higher at 6 weeks. In a different experiment, rats were perfused at 4, 6, and 9 weeks after infection and the number of worms was found to be consistently higher in B rats, by a factor of about 2 at 4 weeks to a factor of about 4 or 6 at subsequent times. Although B rats had more worms than controls even at 9 weeks, a slow drop in their worm burden was noticeable with time in both experiments. Moreover, the size of worms in B rats was smaller than in controls and even 9-week-old worms failed to develop to normal size and appearance and could not be shown to produce fertile eggs. These experiments show a definite involvement of the immune system in the "self-cure" phenomenon, but may at the same time suggest that other non-immune mechanisms are involved in determining the pattern of S. mansoni infection in the rat.


Detection of anti-TNP antibody-forming cells (AFC) with TNP-enzyme and TNP-Fab anti-enzyme conjugates. Peroxidase (PO), alkaline phosphatase, and glucose oxidase, as well as Fab anti-PO, were coupled with varying molar ratios of trinitrophenyl (TNP) hapten. These reagents were evaluated for their ability to detect anti-TNP antibodies in the lymph node cells of Balb/c mice immunized with heavily-substituted TNP45 alkaline phosphate, which gave rise only to anti-hapten antibody-forming cells (AFC). The best results were obtained with lightly-substituted TNP-Fab anti-PO plus PO, TNP-alkaline phosphatase, and TNP-glucose oxidase. These reagents gave strong, specific staining of AFC, and negative background staining. Anti-hapten and anti-carrier AFC could be stained in contrasting colors on the same slide, when immunization was performed with lightly-substituted TNP5.9 alkaline phosphatase. Anti-hapten AFC were detected with TNP-Fab anti-PO or TNP-glucose oxidase, and unsubstituted alkaline phosphatase was used to reveal anti-carrier AFC. The number of AFC detected with these reagents was compared with the number of direct and indirect anti-TNP plaque-forming cells (PFC). At three and five weeks after primary immunization, 40 and 70% more AFC than PFC were detected. These methods can be employed alone, to enumerate anti-TNP AFC and, if desired, anti-carrier AFC; they can also be used in parallel with anti-TNP PFC assays, to determine the fractions of AFC that are not actively involved in antibody secretion.


The micro-membrane-fluorescence test: a new semiautomated technique based on the microtiter system. A semiautomated modification of the membrane immunofluorescence (MF) technique was developed employing units of the Microtiter system. This new rapid technique, which allows the performance of several hundred samples within a few hours is exemplified on a MF test proving the specific reaction to anti-chicken bursa cell sera (ABS) and antichicken thymus cell sera (ATS) on bursa and thymus cells respectively. The possible value of the new method for the further standardization of immunofluorescence is also emphasized.


An immuno-enzymatic assay of cortisol using E. coli beta-galactosidase as label. An enzyme-assay of cortisol was established by using E. coli beta-galactosidase as a marker. The number of cortisol residues per molecule of enzyme and the method of separation of free from antibody-bound enzyme proved to be critical factors for the assessment of the assay. A sensitivity ranging from 100 to 150 pg of cortisol per tube has been achieved. A comparsion with other methods, i.e. competitive protein binding assay, radioimmunoassay and fluorimetry, is reported.


Isolation of lymphocyte subpopulations from rabbit peripheral blood. Comparison of methods based on recovery of rosette-forming cells and on recovery of Ig-bearing cells from a digestible immunoadsorbent. Two methods to obtain lymphocyte subpopulations, defined by specific surface receptors, from rabbit peripheral blood cells were compared as to cell recovery, yield and purity of the obtained fractions: a) the formation of rosettes between lymphocytes and SRBC or SRBC coated with an antigen--antibody--complement complex (D-SRBC), followed by isolation of the rosettes and recovery of the RFC, b) retention of surface-Ig bearing cells on an immunoadsorbent to which antibody to rabbit Ig was covalently attached via a digestible gelatin bridge, with subsequent recovery of the retained cells by the enzymatic digestion of the bridge. Purity of the isolated cell fractions was assessed in all cases by the percentage of cells staining with FITC-labeled goat anti-rabbit Ig. Using the rosette method, all of the RFC could be recovered from rosettes and a very pure, surface-Ig negative, sub-population of cells was obtained: however, the overall number of rosettes formed (SRBC and D-SRBC) was low (8-9% of the nucleated PBC). Very good recoveries and highly enriched cell populations were obtained with the digestible immunoadsorbent, provided certain precautions to minimize cell losses were taken. Thus, 47% of the input cells, representing 90% of the lymphocytes could be recovered; separated cell populations were 96% Ig-positive or, in another experiment, 96% Ig-negative.


Identification of basophilic granulocytes in human leukocyte suspensions subjected to immunofluorescence procedures with anti-IgE. A method is described by which human leukocyte preparations obtained by Ficoll-Isopaque centrifugation are stained and fixed in such a way that the basophilic granulocytes in these preparations can be identified when the latter are subjected to immunofluorescence procedures with anti-IgE. Capping, blocking and absorption experiments confirmed the specificity of the test.


Preparation of conjugated erythrocytes for long term use in hemolytic plaque assays, complement fixation studies, or passive hemagglutinations: a comparative study of several methods. A variety of conjugation procedures, with and without glutaraldehyde stabilized erythrocytes, was employed to couple human IgG to sheep erythrocytes. The efficiency of conjugation, the recovery of cells, and the extent of labeling are noted. These conjugated erythrocytes were then tested for suitability in the hemolytic plaque assay, in a passive hemagglutination test, in a complement assay, and in the detection of rheumatoid factor. Although several methods were useful in a variety of situations, it was found that conjugation using a water soluble carbodiimide (1-ethyl-3-(3-dimethylaminopropyl) carbodiimide) at low temperature (4 degrees C) produced a uniformly acceptable product that was stable and reproducible over a seven week period. The use of this method for a diversity of antigens or haptens is discussed.


Affinity chromatography. A new technique for partial purification of human leucocyte migration inhibitory factor. Human leucocyte migration inhibitory factor (LIF) was subjected to affinity column chromatography on Sepharose-bound alpha-L-fucose. Following the addition of alpha-L-fucose to the running buffer adsorbed lymphokine was eluted almost quantitatively. A minimum of 70-fold purification of LIF was achieved.


A method to study in vitro proliferation of lymphocytes from individual mice on repeated occasions. A micro-method was developed that allowed the proliferation of blood lymphocytes from individual mice incubated with phytohemagglutinin to be measured on repeated occasions. Lymphocytes were isolated from 0.4 ml of heparinized blood by centrifugation through a Ficoll-Hypaque solution. This amount of blood could be taken repeatedly from individual mice and yielded 4-8 X 10(5) lymphocytes. Fifty thousand lymphocytes were incubated with 0.2 ml of medium RPMI 1640 containing 5% human serum in round bottom micro-titer wells. Using this technique lymphocytes from individual mice of the same strain were shown to differ consistently in their responses to PHA.


Antibodies to calf serum as a cause of unwanted reaction in immunofluorescence tests. One potentially important cause of disturbing staining of cell cultures used for immunofluorescence (IF) tests was found to be a specific reaction between calf serum and corresponding antibodies. A simple method utilizing polymerized calf serum as an immunosorbent was used for  the removal of the antibodies causing the undesired reactions. One to three cycles of absorption with the calf serum immunosorbent were highly efficient in removing antibodies to calf serum, whereas the titer of a heterologous antiserum (anti-egg albumin) was only slightly affected by the same procedure. An enzyme linked immunosorbent assay (ELISA) was used for determination of the efficacy of the absorption method.


Detection of human contact sensitivity to dinitrochlorobenzene by the migration inhibiton test. The migration inhibitory factor (MIF) production of peripheral lymphocytes following exposure to dinitrophenylated microsomes derived from both human (DNP-hy-Mic) and guinea pig (DNP-gp-Mic) epidermis was quantitated to detect human contact sensitivity to dinitrochlorobenzene (DNCB). The lymphocytes from nonsensitized subjects did not generate MIF following exposure to either antigen. With DNP-gp-Mic as the antigen, MIF production was noted in only 1 out of 6 DNCB-sensitized subjects and was not significant statistically. With DNP-hu-Mic as the antigen, highly significant MIF production was observed in all 12 sensitized subjects ( p less than 0.0005). In order to confirm MIf production by sensitized lymphocytes following stimulation by DNP-hu-Mic, the subjects were actively sensitized with DNCB and MIF production was assessed before and after sensitization. Remarkable MIf production was noted in the lymphocyte cultures after sensitization, although no significant MIF production was observed before sensitization. MIF production of the sensitized lymphocytes cultured in the presense of DNP-hu-Mic generally correlated well with the results of patch testing, but not with the intensity of the skin test.


Herpes gestationis: fine structural pattern of immunoglobulin deposits in the skin in vivo. IgG and C3 deposits were observed in the basement membrane zone in a case with herpes gestationis. In addition, circulating IgG anti-basement membrane antibodies were found. Electron microscopic immunocytochemical investigations, performed with a peroxidase-antiperoxidase sandwich technique, revealed the in vivo-bound IgG to be localized at the epidermal basal lamina and to exhibit a distribution pattern which is identical to that reported for bullous pemphigoid.


The blood vessels of the skin.

The blood vessels of the skin. During the last 25 years, cutaneous biologists have been particularly interested in abnormal cutaneous vascular patterns, the profusion of capillary anastomoses, the leakiness of venules, clotting, fibrinolysis, and blood viscosity. As a result, the effects of hypoxia and the factors that encourage new vessel proliferation are better understood than before. Only when the biologic behavior of the two extremes of growth from hypoplasia to hyperplasia is studied and compared can the blood supply of a tissue be understood. Hyperplastic tissues are seen in wounds, psoriasis, cancer, and in selected sites of chronic stasis and hypoxia where the vessels are extremely permeable, where blood cells easily escape, and where lymphatics dilate and proliferate. The proliferation of other tissues, such as endothelium, epithelium, mast cells, and probably of locally infective organisms, is also encouraged in hyperplasia. Moreover, fibrinolysis does not occur and fibrin is deposited, the electrostatic charge on the internal vascular surface becomes more positive, and the organ is more vulnerable to subsequent injury. Atrophic or hypoplastic tissues have a reduced cellular turnover and are less hypoxic. The vessels are less permeable, blood cells do not escape, there is only a slight tendency to clot, and fibrinolysis is often increased. Lymphatics are sparse and infection is not a feature. The electrostatic charge on the internal surface of the vessel is negative.


Excelsior: a retrospective view of collagen. The current state of knowledge about the biosynthesis of the collagen molecule and the stabilization of fibers is reviewed. There have been a number of major breakthroughs during the past 25 years which will be described.


Inflammation: a quarter century of progress. This report summarizes the progress of studies on inflammation during the past quarter century; it is a personal overview of the most important and salient accomplishments in this field. Except for  a few references to review articles and textbooks in the field, much of the literature surveyed is not cited.


Developments in delayed-type hypersensitivities: 1950-1975. Significant developments during the last 25 years are discussed and interpreted. The following areas of delayed hypersensitivity are included: the mode of active sensitization to simple allergenic chemicals; evidence for anamnestic responses; cell types and cell-cell interactions via lymphokines; function of skin and lymphatics, and the role of the carrier in initial sensitization to allergenic chemicals; acquired tolerance; transfer factor. Some prognostications for the future are attempted.


Superficial mycoses.

Superficial mycoses. Twenty-five years ago many of the topical remedies for superficial mycoses were irritating, toxic, or allergenic. Total x-ray depilation of the scalp was the accepted mode of therapy for tinea capitis. The introduction of topical nystatin for candidiasis and tolnaftate for dermatophytosis were major advances, but tinea capitis, onychomycosis, and chronic tinea pedis still presented problems. Soon after its introduction in 1958, griseofulvin became the definitive form of therapy for all types of dermatophytosis and played a major role in abolishing large-scale epidemics of tinea capitis in some countries. Recently, haloprogin and the imidazole derivatives, miconazole and clotrimazole, which are topically active against dermatophytes and Candida albicans, have become available. Selective indicator media for isolating dermatophytes are useful diagnostic tools, but quicker methods of diagnosis which require little interpretation are still lacking. Epidemiologic studies in Vietnam again revealed the effects of climate and occlusion on the prevalence, incidence, and severity of superficial mycoses and led to renewed interest in host susceptibility, environment, and prevention of infections.


Genetics and dermatology: a review.

Genetics and dermatology: a review. The development of genetic studies over the last hundred years, in particular as they have affected dermatology, has been reviewed. The clinical application of knowledge derived from other fields has been emphasized, areas of future research have been indicated, and suggestions for cooperation with other disciplines have been offered.


Cutaneous oncology 1950-1975.

Cutaneous oncology 1950-1975. Sociopolitical and economic climates between 1950 and 1975 favored the funding of cancer research, attracted imaginative policy makers, encouraged investigators, and generated new knowledge for clinical application. Important advances during this period include the discovery that DNA alteration is requisite for carcinogenesis and that DNA repair processes are important deterrents in the development of cancer. The better understanding of the histogenesis of epidermal cancer and melanoma has potential clinical value. More easily grasped is the progress made in the therapeutic control of mycosis fungoides and epidermal cancer. Complete control of epidermal cancer is now a foreseeable reality.


Keratinization.

Keratinization. Early studies have already shown that the tonofibrils of malpighian cells consist of a --SH containing fibrous alpha-protein. It was assumed that the highly resistant protective substance, keratin, was formed by the conversion of --SH groups into --S--S--bonds in this protein. This chemical reaction was regarded as the most significant event of the keratinization process. Recent studies show that keratinization proceeds by a synthetic and a degradative stage and that ultimately a complex protective substance is formed. Horny cells become filled with --SH-containing filaments embedded in a --S--S---rich amorphous matrix. This complex is encased by a thickened membrane rendered insoluble by --S--S bonds and an unknown, highly resistant bond. In the stratum corneum, the intercellular space is occupied by bipolar lipids originating from the discharged lamellae of membrane-coating granules.


Cutaneous photobiology: past, present and future. The history and origin of the science of photobiology are reviewed. Interest in the biologic effects of light gradually increased, beginning with the discovery of ultraviolet and infrared radiation early in the 19th century. The basis of experimental photobiology was laid by the studies of Raab and Tappeiner on photodynamic action and the early uses of phototherapy by Finsen and Dorno. The discovery of the association of porphyrins with some light-related skin diseases and of the capability of chemical agents such as coal tar and bergamot to induce phototoxic contact dermatitis resulted in a flurry of clinical investigations leading to better understanding of the processes of phototoxicity and photoallergy. The early epidemiologic studies of Unna and Dubreuilh relating solar radiation exposure to the formation of actinic keratoses and non-melanoma skin cancer were experimentally confirmed in animals by Findlay, Roffo, and Blum. In the most recent quarter century (1950-1975), cellular and molecular photobiology has been refined. The studies on photochemistry of nucleic acid and of damage and repair mechanisms in DNA have set the stage for understanding the basic processes of biologic effects of light and promise the development of useful applications of specifically directed phototherapy and prevention of such light-induced diseases as skin cancer.


Cytokinetics and chemotherapy of psoriasis. The successful treatment of psoriasis with folic acid antagonists during the past 25 years has led to extensive research in the areas of cytokinetics and chemotherapy. In this paper we shall review selected aspects of these topics relevant to the treatment of psoriasis. The effectiveness of methotrexate treatment of psoriasis can be related to both the hyperproliferative cytokinetics of psoriasis and an increased biochemical sensitivity of psoriatic epidermal cells to this drug. Future research goals in chemotherapy of psoriasis include (a) optimizing drug schedules for available drugs; (b) identifying other susceptible biochemical points of selective drug attack; (c) identifying secondary advantages in order to facilitate selective drug action in psoriasis, such as ultraviolet light therapy in combination with a systemic drug; and (d) developing topically effective chemotherapeutic agents. Approaches to research on topical therapy are reviewed with specific reference to animal testing models for psoriasis and percutaneous penetration of topically applied agents.


Trends in electron microscopy of skin.

Trends in electron microscopy of skin. Some trends in electron microscopy of skin have emerged and should be pursued in the future. The fine structure and some basic cellular reaction patterns of epidermal cells are discussed to illustrate the interplay of morphologic, cytochemical, and tracer studies. Intracytoplasmic membranes and secretory granules, lysosomes and endocytic mechanisms, cytomembranes and cell surface specialization are discussed to show how these can be used to arrive at a more meaningful interpretation of structure. Despite all advances, however, a great deal more needs to be done before the details of skin structure are completely elucidated.