Cervical lymphadenitis due to Mycobacterium malmoense in a child: case report and differential diagnosis of cervical lymphadenitis and lymphadenopathies

Cervical lymphadenitis and lymphadenopathy are common in children and may be caused by local or systemic infection. There are a large number of possible etiologies. We report the case of a five-year-old boy with unilateral cervical lymphadenitis who was in good health and showed neither clinical nor laboratory signs of systemic infection. Despite repetitive empirical antimicrobial treatment, the lymph nodes increased in size. After surgical excision Mycobacterium malmoense grew from the lymph node culture. Based on a literature search (Embase and Medline 1980-1992), the most frequent infectious etiologies of pediatric cervical lymphadenitis and lymphadenopathies are summarized. Nontuberculous mycobacterial lymphadenitis is most frequent in children between one and five years of age, and is more common in girls than boys. The treatment of choice is surgical excision. In immunocompetent children treatment with antimycobacterial drugs is not needed even if relapse occurs.     

Isolation and characterization of a novel actin filament-binding protein from Saccharomyces cerevisiae

Cytochrome oxidase (COX) is considered to integrate in a single enzyme two consecutive mechanistically different redox activities--oxidase and peroxidase--that can be catalyzed elsewhere by separate hemoproteins. From the viewpoint of energy transduction, the enzyme is essentially a proton pumping peroxidase with a built-in auxiliary eu-oxidase module that activates oxygen and prepares in situ H2O2, a thermodynamically efficient but potentially hazardous electron acceptor for the proton pumping peroxidase. The eu-oxidase and peroxidase phases of the catalytic cycle may be performed by different structural states of COX. Resolution of the proton pumping peroxidase activity of COX and identification of individual charge translocation steps inherent in this reaction are discussed, as well as the specific role of the two input proton channels in proton translocation.     

Isolation of a novel cytokine from human fibroblasts that specifically inhibits osteoclastogenesis

beta-Carbolines are endogenous inhibitors of monoamine oxidase (MAO). The interaction of nine beta-carboline derivatives and four 3,4-dihydro forms with purified MAO A was investigated. All the compounds tested were reversible competitive inhibitors selective for MAO A, in agreement with previous studies on membrane preparations. The oxidation of kynuramine by MAO A in the presence of the more effective inhibitors showed a lag period before reaching the steady state. In general, the 1-methyl and 7-methoxy substituents increased the potency. Harmine, 2-methylharminium, 2,9-dimethylharminium, and harmaline were the most effective inhibitors of the purified MAO A, with low Ki values of 5, 69, 15, and 48 nM, respectively. The inhibitors interacted with the covalently bound flavin to induce distinct spectral changes, the magnitude of which correlated with the efficacy of the inhibition. The more effective inhibitors could be in situ inhibitors of MAO A.     

Isolation and molecular cloning of a novel type 2 ribosome-inactivating protein with an inactive B chain from elderberry (Sambucus nigra) bark

One of the predominant proteins in the bark of elderberry (Sambucus nigra) has been identified as a novel type 2 ribosome-inactivating protein that exhibits a normal RNA N-glycosidase activity, but is devoid of carbohydrate binding activity. Sequence analysis of the corresponding cDNA clones revealed a striking homology to the previously cloned bark lectins from elderberry, suggesting that the new protein is a lectin-related protein. Molecular modeling of the protein confirmed that its A chain is fully active, whereas its B chain contains two functionally inactive carbohydrate-binding sites. These findings not only demonstrate for the first time the occurrence of a type 2 ribosome-inactivating protein with an inactive B chain, but also offer interesting perspectives for the synthesis of immunotoxins with an improved selectivity.     

A novel presentation of Addison disease: hypoglycemia unawareness in an adolescent with insulin-dependent diabetes mellitus

A 16-year-old boy with insulin-dependent diabetes mellitus (IDDM) and a history of marginal glycemic control had severe hypoglycemia unawareness and a marked decrease in insulin requirement. His counterregulatory hormone response at the time of hypoglycemia suggested adrenocortical and adrenomedullary dysfunction. Further testing confirmed Addison disease. The patient's hypoglycemia unawareness was reversed by glucocorticoid replacement, although the plasma epinephrine response to hypoglycemia remained undetectable.     

Isolation and characterization of a novel gene from human glioblastoma multiforme tumor tissue

The therapeutic application of high-dose interleukin (IL) 2 in human malignancy is limited by severe multiorgan toxicities that are mediated, in part, by tumor necrosis factor (TNF) and IL-1. CT1501R (lisofylline; LSF) is one of several methyl xanthine congeners that inhibit the effects of TNF by the interruption of specific signal transduction pathways. This randomized, placebo-controlled trial was designed to assess the activity of LSF in reducing the toxicities of high-dose IL-2 therapy. Fifty-three patients with metastatic renal cancer or malignant melanoma were treated with i.v. bolus IL-2, 600, 000 IU/kg every 8 h for 5 days (14 doses), followed by 9 days of rest and another 5-day course of IL-2. Patients were randomly assigned to LSF, 1.5 mg/kg i.v. bolus, or placebo every 6 h during IL-2 therapy. All patients were to be treated to individual maximum tolerance of IL-2 at the intensive care unit level of support. The end points for statistical analysis were the number of IL-2 doses administered during the first cycle of treatment (maximum, 28) and the toxicities experienced by each group after the first 8 planned IL-2 doses. There was no difference between the LSF and placebo groups in the mean number of IL-2 doses tolerated in the entire first cycle of therapy (19.6 +/- 5.4 versus 19.5 +/- 5.8, P = 0.86) or in the first or second 5-day course of IL-2. The only significant difference in toxicities occurring through the eighth dose of IL-2 was in the maximum elevation of serum creatinine (mean, 1.7 +/- 0.8 for placebo versus 1.5 +/- 0.6 mg/dl for LSF, P = 0.013). A Monte Carlo analysis of major toxicities over the first 14-dose course of therapy showed a statistically significant difference favoring the LSF-treated group (P = 0.025). LSF was well tolerated, associated only with mildly increased nausea (P = 0.006 after eight IL-2 doses, but not significant for the entire first cycle). The antitumor activity was comparable in both groups (objective responses, 2 of 28 with LSF versus 4 of 24 with placebo). The mean peak plasma concentrations of LSF on days 1, 5, and 19 were 6.24, 3.83, and 5.04 micromol/liter, respectively. In conclusion, with this dose and schedule, LSF did not alter the toxicities of high-dose i.v. IL-2 sufficiently to impact the overall dose intensity of IL-2. Successful IL-2 toxicity modulation may require the use of higher doses of LSF, the development of agents with more potent anti-TNF activity, and/or combined modulating agents that function via distinct mechanisms to interrupt cytokine-mediated signaling.     

Isolation and characterization of a novel trisialoganglioside, GT1a, from human brain

A novel trisialoganglioside has been isolated from normal adult human brain in a yield of 0.6% of the total gangkioside. By graded neuraminidase treatment, mild acid hydrolysis and periodate oxidation analysis, the ganglioside was identified as GT1a having the following structure: NeuAc(alpha, 2-8)NeuAc(alpha, 2-3)Gal(beta, 1-3)GalNAc(beta, 1-4) [NeuAc(alpha, 2-3)]Gal(beta, 1-4)Glc(1-1)ceramide.     

Isolation of a novel gene down-regulated by v-src

We have re-examined the reaction of fast oxidised cytochrome bo with H202 in a stopped-flow spectrophotometer. Monitoring the reaction at 582 nm allows us to observe the formation and decay of a spectroscopically distinct intermediate which accumulates transiently prior to the formation of an oxyferryl species previously characterised in this laboratory (Watmough, N.J., Cheesman, M.R., Greenwood, C. and Thomson, A.J. (1994) Biochem. J. 300, 469-475 [1]). The reaction shows three distinct phases of which the fast and intermediate phases are bimolecular and show a marked pH dependence. Initially these results appeared incompatible with the report that only one equivalent of H202 is required to generate the oxyferryl species (Moody, A.J. and Rich, P.R. (1994) Eur. J. Biochem. 226, 731-737 [21]. However, these data can be reconciled by a branched reaction mechanism whose contributions differ according to the peroxide concentration used.     

Isolation of a novel Kunitz family protease inhibitor in association with Tethya hemolysin from the sponge Tethya ingalli

Aqueous extracts from the New Zealand sponge Tethya ingalli (Hadromerida) displayed potent cytotoxicity in the NCI's 60-cell-line human tumor panel. Fractionation of the extract by ammonium sulfate precipitation, gel filtration, ultrafiltration, and both hydrophobic interaction and reversed-phase chromatography resulted in the isolation of two biologically active proteins. The first protein, Tethya protease inhibitor (TPI), which was purified to homogeneity, inhibited trypsin with an EC50 of 65 nM. TPI had a molecular mass of 11,431 Da, and an isoelectric point of 8.2. A partial N-terminal amino acid sequence determined for TPI showed significant homology with protease inhibitors of the Kunitz family. The second isolated protein displayed potent cytotoxicity, with pronounced selectivity for certain tumor cell lines (e.g., ovarian, renal, CNS, and breast). The latter protein, which had an apparent molecular weight of 21 kDa (SDS-PAGE), also lysed human red blood cells (EC50 of 39 nM) and was similar to a hemolysin previously isolated from the sponge Tethya lycinurium.     

Isolation and characterization of a novel inositol hexakisphosphate binding protein from mammalian cell cytosol

Inositol hexakisphosphate (IP6) is present in most mammalian cells, although its intracellular function is as yet undefined. We find that the total protein fraction from bovine brain cytosol contains a significant level of specific binding for IP6 precipitable with 40% saturated ammonium sulfate. A protein complex has been isolated from this fraction that specifically binds IP6 and is purified about 500-fold over the cytosol. The IP6 binding protein (IP6BP) chromatographs as a single peak of binding activity on a gel exclusion column, with a Stokes radius equivalent to 266 +/- 14 kDa. The IP6BP is a heterooligomeric complex composed of a number of subunits with molecular weights varying from 23,000 to 60,000, as determined by polyacrylamide gel electrophoresis in the presence of sodium dodecyl sulfate (SDS-PAGE). Scatchard analyses of IP6 binding of both the crude ammonium sulfate fraction and the purified complex show the presence of a similar high-affinity binding site (Kd approximately 6.0 nM). Bmax for the purified fraction is 1.8 nmol of IP6/mg of protein or 0.48 mol of IP6 bound/mol of complex. Other inositol polyphosphates, such as inositol 1,3,4,5,6-pentakisphosphate, inositol 1,3,4,5-tetrakisphosphate, and inositol 1,4,5-trisphosphate, are poor competitors for IP6 binding to the purified complex. The purification scheme, when applied to a rat liver cytosol fraction, yields a similar IP6BP. This complex has an apparent size of 512,000 using gel exclusion chromatography and contains an additional protein band with M(r) = 97,000 by SDS-PAGE.(ABSTRACT TRUNCATED AT 250 WORDS)     

Isolation of Corynebacterium pseudotuberculosis from the cervical canal of clinically normal sows

Two isolates from the cervical canal of clinically normal sows were indistinguishable from Corynebacterium pseudotuberculosis in their biochemical properties and genetic homology by DNA-DNA hybridisation. Furthermore, three sheep inoculated with one of the isolates developed typical lesions of caseous lymphadenitis and gave antibody responses specifically to C pseudotuberculosis.     

Isolation of a novel peptide with a unique binding profile from human brain cortex: Tyr-K-MIF-1 (Tyr-Pro-Lys-Gly-NH2)

Tyr-MIF-1 (Tyr-Pro-Leu-Gly-NH2), Tyr-W-MIF-1 (Tyr-Pro-Trp-Gly-NH2), and MIF-1 (Pro-Leu-Gly-NH2) are biologically active peptides previously isolated from brain tissue. We now have used size exclusion chromatography and several consecutive rp-HPLC steps monitored by RIA to isolate a structurally related peptide from human brain cortex with the sequence Tyr-Pro-Lys-Gly-NH2 (Tyr-K-MIF-1). Determination of the sequence, electrospray mass spectrometry, and comparison of its chromatographic behavior with synthetic Tyr-K-MIF-1 confirmed the structure. Unlike Tyr-MIF-1 and Tyr-W-MIF-1, Tyr-K-MIF-1 does not bind to the mu opiate site; unlike MIF-1, Tyr-K-MIF-1 can bind to the Tyr-MIF-1 site. Of these peptides, only Tyr-K-MIF-1 binds to its own site in brain tissue prepared in Tris buffer. Thus, a new member of the Tyr-MIF-1 family of peptides, with a unique profile of binding, has been isolated from human brain cortex.     

The discovery of a novel R-phycoerythrin from an antarctic red alga

A novel biliprotein, named R-phycoerythrin IV, has been discovered. It absorbs blue light better than any other known red algal biliprotein. The protein was found in Phyllophora antarctica, a benthic macroalga, which grows beneath the coastal waters of McMurdo Sound, Antarctica. Fluorescence emission and fluorescence excitation polarization spectroscopy demonstrated that R-phycoerythrin IV behaved as a typical R-phycoerythrin in the functioning of energy migration and has an emission maximum at 577 nm. The circular dichroism (CD) spectrum of the chromophores was compared with visible absorption spectrum, and both were deconvoluted. This process showed the energy states of various individual chromophores. The molecular weight of the protein suggested a alpha6beta6gamma polypeptide structure, and far UV CD studies revealed polypeptides with highly alpha-helical secondary structures. Dynamic light scattering indicated that the protein had a 5.54 nm radius, and its shape was nonspherical. R-phycoerythrin was also purified from a second benthic Antarctic red alga, Iridaea cordata. Its spectroscopic properties were similar to those of some R-phycoerythrins from nonpolar regions. The unique spectroscopic properties of R-phycocerythrin IV may help enable the alga to occupy its niche deeper in the water column than the red alga that has the typical R-phycoerythrin.     

Isolation of a novel RFamide peptide from the midgut of the American cockroach, Periplaneta americana

In insects (FM)RFamide-immunoreactive endocrine cells are ubiquitously present in the midgut, but the identity of the peptide(s) produced by these cells is unknown. The major RFamide-immunoreactive peptide from the midgut of the American cockroach, Periplaneta americana, was isolated and identified as Ala-Asn-Arg-Ser-Leu-Arg-Leu-Arg-Pheamide. This is a novel member of an arthropod peptide family, previously known only from mosquitoes and horseshoe crabs. Its abundance in the midgut suggests that it plays an important function in digestion.     

Renal adenocarcinoma with lung metastasis in an adolescent

OBJECTIVE: The finding of a renal adenocarcinoma in a young patient is a rare occurrence. We report on a case of renal adenocarcinoma with lung metastases arising in an 18-year-old girl with non-specific symptoms and normal blood chemistry and urinary analysis. RESULTS AND CONCLUSION: We underscore the aggressive nature of the tumour which grew asymptomatically, giving rise to lymph node and pulmonary metastases.     

Gastric carcinoma with ovarian metastases in an adolescent

Gastric carcinoma represents only 0.05% of malignant paediatric gastrointestinal tumours. This condition is associated with a poor prognosis because its rarity and non-specificity of the symptoms usually delay its diagnosis. The authors present a 16-year-old girl with a poorly differentiated gastric carcinoma (signet-ring-cell type) with bilateral ovarian metastases who died of peritoneal carcinomatosis despite extensive surgery and chemotherapy. The epidemiological, clinical, and pathological features of gastric carcinoma in childhood are discussed.     

Isolation, characterization and synthesis of a novel paradaxin isoform

Recently our laboratory found that tolerance to morphine-induced antinociception could be completely reversed with intracerebroventricular (i.c.v.) administration of a protein kinase A inhibitor, whereas intrathecal (i.t.) administration of the inhibitor produced hyperalgesia in morphine-tolerant mice. In the experiments described here, we sought to characterize further the role of phosphorylation events in supraspinal versus spinal opioid-mediated pain pathways and how such events might be involved in the development of antinociceptive tolerance. Two phosphatase inhibitors were administered centrally to determine whether they affected morphine-induced antinociception in naive or chronically morphine-treated mice. By the i.c.v. route, okadaic acid enhanced morphine-induced antinociception in tolerant mice and produced toxicity by the i.t. route. The calcineurin inhibitor ascomycin had no effect on antinociception following acute or chronic morphine treatment. These results suggest that increased activity of protein phosphatase types 1 and/or 2A in the brain may contribute to the development of morphine tolerance.