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D-Glucosamine is toxic to several malignant cell lines and in vivo tumors at concentrations that have little effect upon normal host tissues. Evidence is presented to support the hypothesis that cellular membranes may be the primary targets of glucosamine's tumoricidal activity. Treatment of rat C6 glioma cells with a cytotoxic concentration of glucosamine (20 mM) caused fragmentation of rough endoplasmic reticulum, proliferation of Golgi complexes, evagination of outer nuclear and mitochondrial membranes, and the accumulation of membranous vacuoles and lipid droplets in the cytoplasm. These changes were detected within the first 3 hr after treatment of cultures with glucosamine and became increasingly severe until cell lysis occurred between 24 and 48 hr of treatment. The cytotoxicity of glucosamine was potentiated by the local anesthetic lidocaine, and by other membrane-active drugs, at concentrations that were growth inhibitory but nonlytic. Most of these drugs possessed local anesthetic activity and inhibited glioma sterol synthesis. Within the same period of time required for ultrastructural changes in cellular membranes, glucosamine inhibited the incorporation of [2-(14)C]acetate into sterols and into an unidentified 400-dalton lipid that migrated close to sterols on thin-layer chromatograms. This inhibition was potentiated by lidocaine and increased over the same range of D-glucosamine concentrations that led to increased cell toxicity after a 48-hr treatment. These findings suggest that the effects of glucosamine upon cellular membranes may be central to its tumoricidal activity and that glucosamine, in combination with membrane-active drugs, may be useful in the treatment of certain types of tumors, particularly those of the central nervous system.

We report on the cases of two patients in whom linear perilesional hypopigmentation and atrophy developed after intralesional injection of corticosteroids for treatment of keloids. Evaluation of our patients and the previously reported cases showed that perilesional linear atrophy or hypopigmentation (or both) is a distinct complication after intralesional or intraarticular administration of corticosteroids and is probably due to lymphogenous spread of the steroid suspension.

Shortly after reaching confluency, canine MDCK cells enter a prolonged state of basal growth with doubling times of 200-300 hours. These values are similar to those commonly exhibited by in vivo solid tumors at clinically relevant sizes. By comparison with rapidly growing sparse density cultures, the postconfluent monolayers displayed a pronounced resistance to deazauridine, deoxyspergualin, and 5-fluorouridine. Drug concentrations required for unit levels of effect increased from several fold to several orders of magnitude as cells entered high density basal growth. This high density chemoresistance was observed for both growth inhibition and cytotoxicity, but was much more pronounced with the former. Dose-response curves were biphasic, suggesting that growth inhibition and cytotoxicity may be mediated by different mechanisms of drug action. The pronounced chemoresistance of postconfluent MDCK monolayers is similar to that encountered with many clinical solid neoplasms. It suggests that postconfluency monolayers, like multicellular spheroids and cellular multilayers, may provide better in vitro models of solid tumor chemosensitivity than subconfluent monolayer and suspension cultures.

A 56-year-old woman developed Favre-Racouchot syndrome involving her face and scalp primarily at the sites of x-ray irradiation for therapy of an astrocytoma. The patient had not had comedones prior to radiotherapy and did not have a history of excessive sun exposure. The patient showed an excellent response to topical retinoic acid gel. To the best of our knowledge, this is the first case of Favre-Racouchot syndrome developing after radiation therapy to be reported in the literature; its pathogenesis is discussed in this paper.

Although leg ulcers have many causes, the most common etiology is venous or arterial insufficiency. The prime objective in the evaluation of leg ulcers is to uncover the precipitating pathologic process, and the first goal of therapy is to correct or control this underlying condition. Simple laboratory tests and office procedures are adjuncts in diagnosis. Topical care of leg ulcers includes gentle debridement, use of wet dressings and, in selected cases, skin grafting.

The authors report a study of catecholamine levels in the spinal cords of monkeys following a 300 gm-cm blow to the midthoracic spinal cord. There was a progressive decrease in norepinephrine (NE) activity as a first-order process with a half-life of 6.4 hours. The NE activity in injured tissue never exceeded control levels, which remained unchanged both above and below the injured segment. Dopamine activity remained unchanged in the injured tissue as well as in control segments above and below the area of trauma for the first hour.

The effect of preferential delivery of estradiol (E2) into the brain on both the hypothalamic-pituitary-ovarian axis and weight gain was studied in female rats. When E2 was coupled to a lipoidal dihydropyridine-pyridinium carrier, the resulting carrier E2 complex (CE), upon a single intravenous administration to cycling female rats, caused a dose-dependent inhibition of ovulation which lasted 3 times longer than with uncoupled E2. The dose of CE that delayed ovulation for 4 days was one twentieth the amount of E2 needed to produce the same effect. Studies in ovariectomized (OVEX) rats indicated that the prolonged ovulation-blocking action of CE appeared to be related to a sustained storage and release of E2 in the brain, which in turn suppressed the release of hypothalamic luteinizing hormone-releasing hormone (LHRH) and pituitary luteinizing hormone (LH). Upon single intravenous administration in pubertal female rats, CE caused a dose-dependent reduction of body weight gain for a minimum period of 28 days. The inhibitory action of CE on body weight gain was more potent and longer lasting than that of E2 in pubertal rats. When administered in OVEX rats, CE produced a loss of body weight that lasted significantly longer than that produced by uncoupled E2 in these rats. These results suggest that the biological action of E2 can be potentiated by this novel chemical delivery system.

Generation time analysis by time lapse cinematography is an important method for investigating cellular senescence in culture, but its interpretation is complicated by several types of bias and artifact, including small sample size, cut-off bias, changes in global growth rate, and phase of the population growth cycle. When these factors are considered, interpretation of the data base used by previous investigators changes considerably, and does not reveal any differences in growth behavior between middle and late passage WI-38 cells. Nor does it support the transition probability theory either of cell cycle transit or of culture senescence.

Previous publications from this laboratory have demonstrated that agents such as methotrexate (MTX), 5-fluorodeoxyuridine (FUdR), trimethoprim, and D-glucosamine (D-GlcN), which are known to inhibit thymidylate synthesis, can augment human NK activity in vitro. Furthermore, this augmentation was inhibited by exogenous thymidine (TdR) at concentrations of 10(-6) to 10(-7) M. In this report, underlying mechanisms of action of FUdR, D-GlcN, and IFN are compared. Each of these agents increased the lytic activity of effector cells bound to targets but did not increase the percentage of conjugates formed. The augmentation could be induced in a population highly enriched for NK cells (Leu-1 lb positive in phenotype). FUdR and D-GlcN could not induce any augmentation in a Leu-1 lb-negative subpopulation whereas IFN could induce significant lytic activity. alpha-Amanitin, an inhibitor of RNA polymerase II, blocked the activation of NK activity by all three reagents; hence gene expression was required. Comparison of [35S]methionine-labeled proteins by two-dimensional gel electrophoresis revealed that six new proteins were induced in IFN-treated cells. Three of these were similar in pI and molecular weight to the newly synthesized proteins in the D-GlcN-treated cells. One protein was synthesized in increased amounts in the FuDR-treated cells and it was not common to either of the other treatments. Evidence to date is consistent with the hypothesis that separate mechanisms underlie the activation of NK cells by IFN and thymidylate synthesis inhibitors, although the existence of a final common pathway for all NK response modulators cannot be excluded at the present time.

We have developed an immunoradiometric assay for IgE antibodies to Staphylococcus aureus (Staph IgE-Ab) which uses purified cell walls (PCW) from the Wood 46 strain of S. aureus as an immunosorbent. We compared Wood 46 PCW and whole organisms (WO) as immunosorbents for Staph IgE-Ab by performing tests on sera from patients with atopic dermatitis (AD) or the hyperimmunoglobulin E syndrome (hyper IgE syndrome). Sera with Staph IgE-Ab demonstrated dose-dependent binding to PCW and WO, but the ratio of specific to non-specific binding was much greater with PCW. Mean non-specific binding to WO was greater than to PCW, 5% versus 2%; and non-specific binding to WO varied directly with the serum concentration of IgE. Results of tests on patients' sera indicated that PCW are required in screening assays for Staph IgE-Ab to avoid false positive results caused by high levels of non-specific binding to WO.

Two families with anetoderma are described. Unlike previous reports of familial anetoderma, the disease process seemed to be limited to the skin, and there were no associated ocular, gastrointestinal, or orthopedic anomalies in the affected patients or in any other family members. Although infrequently reported, anetoderma may occur in families, and patients must be examined for associated systemic abnormalities for a thorough assessment of their skin disorder.

Thymidine kinase activity in rat C6 glioma cells is inhibited by 50 to 70% after 4 hr incubation with 20 mM D-glucosamine. The inhibition is uncompetitive with respect to thymidine, reducing both the apparent Km and Vmax of the enzyme. The inhibition does not appear to be caused by the reversible combination of the enzyme with a cytoplasmic inhibitor, including D-glucosamine and its metabolites. The addition of D-glucosamine or its metabolites to cell-free thymidine kinase produced an inhibition which differed quantitatively and qualitatively from that which resulted from treatment of intact cells with D-glucosamine. The presence of a reversible cytoplasmic inhibitor of the enzyme was also excluded by mixing experiments. D-Glucosamine inhibited the incorporation of labeled uridine and amino acids into acid-precipitable material. The magnitude of inhibition of thymidine kinase activity and amino acid incorporation by D-glucosamine was comparable to that produced by cycloheximide, suggesting that the inhibition might arise from interference with enzyme synthesis. However, whereas the kinetics of recovery of amino acid incorporation from inhibition was rapid, thymidine kinase activity was depressed for at least 6 hr after drug washout. The results presented are best explained by assuming either that two forms of thymidine kinase are present in rat C6 cells and are differently affected by D-glucosamine or that D-glucosamine acts by two separate mechanisms to inhibit a single form of the enzyme.

We determined the prevalence of IgE antibodies to Staphylococcus aureus by optimized immunoradiometric assay methods in serum specimens from 69 patients with atopic dermatitis. All patients had positive aerobic cultures for S aureus from skin. Significant binding attributable to IgE antibodies was noted in three of 25 patients with atopic dermatitis and superimposed impetiginization or pustules, but antibodies were not detected in the remaining 44 patients whose lesions were colonized with S aureus. By comparison, IgE antibodies to S aureus were uniformly present in high titer in serum samples from patients with the hyperimmunoglobulin E syndrome. We conclude that most patients with atopic dermatitis do not have detectable levels of IgE antibodies to S aureus.

D-Glucosamine has been shown in animal studies to have selective tumoricidal activity. In human cancer patients, preliminary data indicate that natural killer (NK) activity is increased secondary to D-glucosamine infusion. The present study examined the effect of D-glucosamine on several in vitro indices of human immune responsiveness including NK cell activity and T and B cell mitogenesis. NK activity in normal human peripheral blood mononuclear cells was significantly elevated in the presence of 10(-4) and 5 X 10(-4) M D-glucosamine. The increment in NK activity was mediated by nonadherent effector cells and was not due to an increased susceptibility of target cells. Analysis by single-cell assay indicated that the number of effector/target conjugates was not increased but that there was increased lytic activity of the NK cells. Previous studies from our laboratory indicated that several drugs which perturb thymidine (TdR) metabolism were effective in enhancing the in vitro NK activity. As D-glucosamine has been observed to alter cellular pyrimidine nucleotide pools and TdR metabolism in tumor cells, the effect of exogenous TdR was assessed on D-glucosamine-stimulated NK activity. Exogenous TdR inhibited D-glucosamine-induced augmentation of NK activity in a dose-dependent manner and completely abrogated the drug response at 10(-7) M. Similar experiments indicated that TdR did not affect the interferon-induced augmentation of NK activity in concentrations up to 10(-4) M. Although human NK activity was enhanced by D-glucosamine in vitro, there was no change induced in T or B lymphocyte mitogenesis.(ABSTRACT TRUNCATED AT 250 WORDS)

A 3-year-old boy with nonbullous congenital ichthyosiform erythroderma with a four-month history of scabies was treated with a single dose of lindane cream. In a 48-hour period, he developed nausea and vomiting and also suffered from an epileptiform convulsion and muscular spasms. Seventy-two hours after application of the cream, his blood lindane level was 54 ng/mL. Caution should be exercised when using lindane in patients with compromised epidermal barrier function.

A case is reported of a forty-nine-year-old black man in whom numerous skin-colored papules and verrucoid plaques had developed on his penis over the course of fifteen years. He did not seek medical attention, and some of the lesions had become quite large. The initial clinical impression was condyloma acuminatum, and prior to therapeutic intervention histologic evaluation revealed findings diagnostic of seborrheic keratosis. Seborrheic keratoses should be considered in the differential diagnosis of penile lesions especially because of clinical similarities to condylomata acuminata.

Sarcoma 180 monolayers spontaneously shed single cells and small multicellular aggregates into the surrounding medium to produce a dual population of floating and substratum-attached cells. Shedding was a motility-associated event that occurred when cells attempted to migrate over one another. It resulted from a combination of cell shape change and active motility, which increased sensitivity to fluid shear dislodgement by reducing a cell's surface area of adhesive contact and increasing strain tension at its adhesive contact points. Shedding occurred at all phases of the cell cycle. Extracellular matrix but not conditioned medium enhanced the floating subpopulation by slowing the kinetics of reattachment to plastic and cellular substrate. Although sarcoma 180 cells are anchorage independent in the sense that they grow readily in single cell suspension, they nevertheless exhibited anchorage modulation of their cell cycle. Short periods in suspension produced a mild G1 accumulation, whereas longer periods of anchorage deprivation led to a mild G2 accumulation which appeared to result from an interference with cytokinesis.

The amino sugar D-glucosamine possesses antitumor activity which is thought to depend in part upon its ability to impair cholesterol biosynthesis and damage cellular membranes. The present study examined the effect of glucosamine on acetate utilization for lipid and sterol synthesis in rat C6 glial tumor cells. At cytotoxic concentrations, the amino sugar inhibited [14C]acetate incorporation into nonesterified sterols and lipids but increased the flow of label into cholesteryl esters. A comparison of the rates of acetate utilization for glucosamine metabolism (N-acetylation) and sterol and lipid synthesis suggested that glucosamine might act by competing for a common cytosolic pool of acetyl CoA. The inhibition of lipid and sterol synthesis, however, remained constant over a wide range of extracellular acetate concentrations. These results suggest that, if glucosamine acts by restricting the supply of acetate for these biosynthetic processes, it probably inhibits a step prior to the formation of acetyl CoA. Alternative mechanisms are discussed.

Two separate control processes govern the cell cycle of rat C6 glioma cells. In subconfluent cultures growth inhibition is caused by cell contact interactions and the cell cycle is regulated primarily by changes in the duration of S phase. During advanced multilayering, medium depletion becomes the primary mechanism of growth inhibition and causes a pronounced G1 accumulation. Contact modulation acts by altering the velocity with which cells progress through the cell cycle, while depletion causes cycle arrest.

Fourteen dental amalgam alloys were used in this study. After 1 year, amalgam restorations were evaluated for fracture at the margins. The clinical results indicated that Dispersalloy, Indiloy, a high-copper blend by Syntex, Cluster, and Unison had the least marginal failure. This was followed by Premalloy, Cupralloy, Tytin, Cupralloy ESP, and Contour. Velvalloy and Sybraloy and Orosphere II were the third major grouping with Summalloy having the most fracture at the margins. Different batches of the same alloy performed similarly. The correlation between creep and ridit means (fracture at the margins) was not found to be statistically significant.

Quadrat analysis was used to investigate the spatial distribution of seven mammalian cell lines in culture. The number of cells in replicate unit areas of the culture was determined, and the variance to mean ratio used as an index of random and nonrandom spatial distribution. Only mouse SV3T3 cells distributed themselves randomly throughout the entire culture growth cycle. The remaining six lines all assumed a nonrandom distribution at some point in their growth cycles. Mouse L929 cells displayed avoidance behavior, and spaced themselves at regular intervals in a uniform spatial distribution. The five remaining lines (mouse S180, rat C6, hamster CHO, canine MDCK, and human BeWo) formed multicellular clusters, and were distributed aggregatively rather than randomly. Random walk migration can account for the random distribution of SV3T3 cells. Random walk combined with contact inhibition of movement provides a satisfactory explanation for the uniform distribution of L929 cells. Random walk and contact inhibition are incompatible with cell clustering, however. Thus other mechanisms of motility or adhesiveness must contribute to cell clustering. It is possible that random walk and contact inhibition may be less common components of cell movement than generally assumed.

A 16-year old boy with a 10-year history of circulating eosinophilia was diagnosed having the hypereosinophilic syndrome (HES) based upon the exclusion of other disorders. Eight years after the onset of his condition, he had a subcutaneous staphylococcal abscess followed by lymphangitis, rare clinical features of HES. As measured by radioallergosorbent techniques, there were significantly high serum levels of IgE antibodies to Staphylococcus aureus. The clinical significance of these antibodies is unknown, but their production may be due to persistent antigenic stimulation.

The amino sugar, D-glucosamine, inhibits the preformed route of thymidine metabolism in rat C6 glioma cells. This inhibition results from a concatenation of several distinct effects, including the inhibition of thymidine uptake, the reduction of thymidine phosphorylation, and an increased leakage of thymidine to the extracellular space. Each of these effects, while ostensibly small in magnitude, is significant and contributes to the observed inhibition of acid-precipitable thymidine incorporation by D-glucosamine. These effects of D-glucosamine, together with its known ability to reduce uridine nucleotide pools, may contribute to its toxicity toward certain experimental animal tumors.

Kawasaki syndrome (KS) is an idiopathic, acute, febrile, exanthemous illness that primarily affects infants and children. We describe a 20-year-old black woman who fulfilled the clinical criteria for the diagnosis of KS and excluded other possible causes. In addition, we reviewed data on 21 patients with adult KS reported in the English literature and accepted ten cases as representing this syndrome. The epidemiologic, clinical, laboratory, and pathologic features of the 11 cases representing adult KS are discussed. Although the initial reports of adult KS in the United States may have actually represented toxic shock syndrome, the occurrence of KS in adults should be acknowledged.

Two unrelated patients had numerous palmoplantar "music box spine" keratotic plugs and pits of 11 and 13 years' duration. Histologic examination revealed a compact column of parakeratosis resembling that of a cornoid lamella of porokeratotic conditions. Ultrastructurally in clinically affected skin, the stratum corneum contained numerous variable-sized pyknotic nuclei, and cells in the stratum granulosum contained fewer keratohyalin granules. The ultrastructural findings differed from those of porokeratosis of Mibelli and disseminated superficial actinic porokeratosis. The proper nosologic designation should be punctuate porokeratotic keratoderma.

A 20-year old man with steatocystoma multiplex and pseudofolliculitis barbae was treated unsuccessfully with oral isotretinoin. Consistent with findings from previous reports, treatment with isotretinoin should be reserved for patients with steatocystoma multiplex suppurativum.

The typical infection with Sporothrix schenckii is characterized by superficial cutaneous nodules occurring along the lines of lymphatic drainage of the limbs. Review of the medical records at the Mayo Clinic from 1937 to the present disclosed 58 patients with sporotrichosis. Eleven patients had evidence of extracutaneous infection. Eight of the patients were men whose ages ranged from 25 to 71 years; the ages of the three women were 54, 64, and 67. Seven patients had predominantly joint involvement, with the knee and wrist joints being most often infected. Other sites of infection included the mandible and ethmoid sinuses. Three patients had disseminated systemic infection, and one patient died within a year of the initial diagnosis. Nine patients had been in good health before the infection, although five patients were taking systemic corticosteroids before their infection was diagnosed. Therapies included supersaturated potassium iodide, amphotericin B, and 2-hydroxystilbamidine isethionate. Although most commonly seen as a cutaneous disease, sporotrichosis is a potentially disseminated infection with life-threatening consequences.