International guidelines for the diagnosis and management of hereditary haemorrhagic telangiectasia.

BACKGROUND: HHT is an autosomal dominant disease with an estimated prevalence of at least 1/5000 which can frequently be complicated by the presence of clinically significant arteriovenous malformations in the brain, lung, gastrointestinal tract and liver. HHT is under-diagnosed and families may be unaware of the available screening and treatment, leading to unnecessary stroke and life-threatening hemorrhage in children and adults. OBJECTIVE: The goal of this international HHT guidelines process was to develop evidence-informed consensus guidelines regarding the diagnosis of HHT and the prevention of HHT-related complications and treatment of symptomatic disease. METHODS: The overall guidelines process was developed using the AGREE framework, using a systematic search strategy and literature retrieval with incorporation of expert evidence in a structured consensus process where published literature was lacking. The Guidelines Working Group included experts (clinical and genetic) from eleven countries, in all aspects of HHT, guidelines methodologists, health care workers, health care administrators, HHT clinic staff, medical trainees, patient advocacy representatives and patients with HHT. The Working Group determined clinically relevant questions during the pre-conference process. The literature search was conducted using the OVID MEDLINE database, from 1966 to October 2006. The Working Group subsequently convened at the Guidelines Conference to partake in a structured consensus process using the evidence tables generated from the systematic searches. RESULTS: The outcome of the conference was the generation of 33 recommendations for the diagnosis and management of HHT, with at least 80% agreement amongst the expert panel for 30 of the 33 recommendations.

Functional organelles in prokaryotes: polyhedral inclusions (carboxysomes) of Thiobacillus neapolitanus.

The polyhedral inclusions of Thiobacillus neapolitanus have been isolated; they contain ribulose diphosphate carboxylase.

Rapid conversion to high xanthine oxidase activity in viable Kupffer cells during hypoxia.

It has been widely postulated that the central mechanism of hepatic reperfusion injury involves the conversion, during ischemia, of the enzyme xanthine dehydrogenase (XDH) to its free radical-producing form, xanthine oxidase (XOD). However, this theory has been questioned because (a) XDH to XOD conversion in whole liver occurs very slowly; (b) the cellular distribution of XDH/XOD is unclear; and (c) the direct demonstration of XDH to XOD conversion in viable cells is lacking. In this paper, we address all three issues by measuring XDH to XOD conversion and cell viability in purified populations of hepatic endothelial cells (EC), Kupffer cells (KC), and hepatocytes (HEP). Although XDH/XOD activity on a cellular basis was greater in hepatocytes (0.92 +/- 0.12 mU/10(6) cells) than ECs (0.03 +/- 0.01) or KCs (0.12 +/- 0.04), XDH + XOD specific activity was similar in all three cell types (HEP 1.85 +/- 0.10 U/g protein; EC 1.69 +/- 0.54; KC 2.30 +/- 0.22). Over 150 min of warm (37 degrees C) or 24 h of cold (4 degrees C) hypoxia, percent XOD activity increased slowly in ECs, from 21 +/- 2% (basal) to 39 +/- 3% (warm) and 49 +/- 5% (cold) and in HEPs (29 +/- 2% to 38 +/- 3% and 49 +/- 2%), but converted significantly faster in KCs (28 +/- 3% to 91 +/- 7% and 94 +/- 4%). The dramatic changes in Kupffer cell XOD during cold hypoxia occurred despite only minor changes in cell viability. When hypoxic KCs were reoxygenated after 16 h of cold hypoxia, there was a marked increase in cell death that was significantly blocked by allopurinol. These data suggest that significant conversion to the free radical-producing state occurs within viable KCs, and that Kupffer cell XOD may play an important role in mediating reperfusion injury in the liver.

Isolation and partial characterization of a 67Ga-binding glycoprotein from Morris 5123C rat hepatoma.

A Glycoprotein, particularly high in tumors, has been extracted from Morris 5123C rat hepatomas and purified. The compound constitutes a major binding component for 67Ga in this hepatoma. It has a molecular weight of approximately 45,000. Its molecular weight was determined by sodium dodecyl sulfate:polyacrylamide gel electrophoresis and by Sephadex G-200 superfine gel filtration. The steps involved in its extraction and purification include ultrafiltration, gel filtration through Sephadex G-200 superfine, ion-exchange chromatography on diethylaminoethyl Sephadex A-50, and hydroxylapatite chromatography. The homogeneity of the compound was established by gel electrophoresis. The NH2-terminal residue, the percentage of nitrogen, the nonamino carbohydrate content, and the amino acid composition are reported.

A quantitative study of the subcellular localization of 67Ga.

This paper describes a quantitative procedure for the isolation of 67 Ga-binding granules (GBG) from normal rat liver and Morris 5123C hepatoma homogenates by a combination of rate and isopyknic density gradient zonal centrifugation. Another class of GBG has been found that is much smaller than the lysosomal GBG we have previously described. These smaller particles, or microvesicles, bind the largest portion of the 67Ga found in the hepatoma whereas, in the liver, the GBG lysosomes are the major binding component. Previously, we had shown that considerably more 67Ga is taken up in hepatoma than in liver (as percentage of administered dose per g of tissue). The preferential association of 67Ga with these microvesicles in the 5123C hepatoma may be indicative of a basic difference between normal and malignant tissue.

Correlation between clinical and MRI assessment of depth of invasion in oral tongue squamous cell carcinoma.

BACKGROUND: Neck metastasis is the most important prognostic factor in oral cavity squamous cell carcinomas (SCC). Apart from the T- stage, depth of invasion has been used as a highly predictable factor for microscopic neck metastasis, despite the controversy on the exact depth cut off point. Depth of invasion can be determined clinically and radio logically. However, there is no standard tool to determine depth of invasion preoperatively. Although MRI is used widely to stage the head and neck disease, its utility in depth evaluation has not formally been assessed. OBJECTIVE: To compare preoperative clinical and radiological depth evaluation in oral tongue SCC using the standard pathological depth. To compare clinical and radiological accuracy between superficial (<5 mm) vs. deep invaded tumor (≥5 mm) METHODS: This prospective study used consecutive biopsy-proven oral tongue invasive SCC that presented to the University health network (UHN), Toronto. Clinical examination, radiological scan and appropriate staging were determined preoperatively. Standard pathology reports postoperatively were reviewed to determine the depth of invasion from the tumor specimen. RESULTS: 72 tumour samples were available for analysis and 53 patients were included. For all tumors, both clinical depth (r = 0.779; p < 0.001) and radiographic depth (r =0.907; p <0.001) correlated well with pathological depth, with radiographic depth correlating slightly better. Clinical depth also correlated well with radiographic depth (r = 0.731; p < 0.001). By contrast, for superficial tumors (less than 5 mm on pathological measurement) neither clinical (r = 0.333, p = 0.34) nor radiographic examination (r = - 0.211; p = 0.56) correlated with pathological depth of invasion. CONCLUSION: This is the first study evaluating the clinical assessment of tumor thickness in comparison to radiographic interpretation in oral cavity cancer. There are strong correlations between pathological, radiological, and clinical measurements in deep tumors (≥5 mm). In superficial tumors (<5 mm), clinical and radiological examination had low correlation with pathological thickness.

Changes in glutathione in intact erythrocytes during incubation with penicillamine as detected by 1H spin-echo NMR spectroscopy.

1H spin-echo NMR spectroscopy was used to study changes in glutathione status in intact erythrocytes. The concentration of glutathione in suspensions of erythrocytes in 2H2O saline is significantly different in cells from patients with rheumatoid arthritis than in controls from normal healthy volunteers. It is observed that the methods of separation and suspension affect lactate metabolism in red cells. Incubation of erythrocytes with solutions of the therapeutic agent D-penicillamine in 2H2O saline produced a change in glutathione resonances which is indicative of an increase in diglutathione concentration. Signals from the methyl groups of penicillamine decreased at a commensurate rate. Incubation of normal cells with plasma from patients with rheumatoid arthritis who had the same blood group as the normal volunteer indicated a much larger fall in glutathione signal with plasma from a patient treated with penicillamine than from a patient on non-steroidal anti-inflammatories.

Ultrastructure and physical characteristics of proplastids and chloroplasts isolated by zonal centrifugation from Euglena gracilis var. bacillaris.

Proplastids at various stages of development and mature chloroplasts were isolated by zonal centrifugation in a double-sigmoid, sucrose-sorbitol gradient from Euglena gracilis var. bacillaris grown in darkness and subjected to various periods of illumination with white light. Computer programs were used in characterizing physical properties of the isolated organelles and in predicting the location of plastids of different developmental stages in the gradient. The technique permits computation of these parameters in any gradient material. The convoluted nature of the early developing plastids, revealed by electron microscopy, suggests that the frictional coefficients should be taken into account in describing the physical constants of the organelles, since their sedimentation coefficients are lower than would be expected for spherical particles of the same size and density.

Antiinflammatory effects of some copper complexes.

Copper complexes of a range of ligands have been prepared and evaluated for antiinflammatory activity and irritancy after oral, subcutaneous, and local administration in rats and guinea pigs. The antiinflammatory activities were found to depend on the species used and the route of administration. When nonantiinflammatory ligands were used, the response was generally dose dependent. With D-penicillamine and when the ligands were themselves antiinflammatory in animal models of inflammation--as was the case with flufenamic acid, levamisole, aspirin, L-histidine, and 2-amino-2-thiazoline--differences in antiinflammatory activity were observed between the copper complexes and the free ligands. In some cases, the copper complexes were the more effective. There was a weak correlation between local (subplantar) irritation and the dose of copper but, for four compounds studied in more detail, the response in the local subplantar test and degree of antiinflammatory activity were not related, suggesting that the action of copper is not solely by a counterirritant mechanism. No obvious differences between the activities of copper(I) and copper(II) compounds were observed, suggesting that a common metabolite may be involved in the antiinflammatory action of copper.

Activation of the hypothalamic-pituitary-adrenal axis differentially affects the anti-mycobacterial activity of macrophages from BCG-resistant and susceptible mice.

The effect of hypothalamic-pituitary-adrenal (HPA) axis activation and exogenous glucocorticoids on the ability of splenic macrophages to control the growth of Mycobacterium avium was evaluated. We found that activation of the HPA axis by restraint stress or the addition of corticosterone increased the susceptibility of macrophages from mice that are innately susceptible to the in vivo growth of M. avium. In contrast, the ability of macrophages from innately resistant, congenic mice to control the growth of M. avium was not affected by HPA activation or the addition of corticosterone. The effect of restraint and of corticosterone on macrophage function was abrogated by either treating mice with the glucocorticoid receptor antagonist RU486 or the addition of the drug to cultures of macrophages. Activation of the HPA axis as well as the addition of corticosterone to cultures of macrophages resulted in a suppression of the production of tumor necrosis factor (TNF)-alpha and of reactive nitrogen intermediates by macrophages from both strains of mice. The lack of effect of HPA activation and of corticosterone on the mycobacterial resistance of macrophages from BCG-resistant mice, while at the same time suppressing the production of reactive nitrogen intermediates, appears to rule out a role for this antimicrobial pathway in innate resistance to mycobacterial growth.

FACTORS AFFECTING THE KINETICS OF INTRA-AND EXTRACELLULAR CADMIUM UPTAKE BY THE MOSS RHYTIDIADELPHUS SQUARROSUS.

The responses of field-grown and cultured shoots of a moss, Rhytidiadelphus squarrosus (Hedw.) Warnst, to Cd were investigated. Respiration was not affected, but photosynthesis was progressively reduced with increasing Cd uptake. Field-grown shoots of two morphologically distinct populations differed in their photosynthetic responses to Cd. This could be related to different rates of Cd uptake by the two populations. During storage of Cd-treated shoots, movement of extracellular Cd to an intracellular location occurred, and this was reflected in altered photosynthetic rates. The maximum extracellular uptake capacity and affinity for Cd were identical for the two populations of field-grown shoots. Field-grown material from the two populations differed significantly in their intracellular Cd uptake characteristics, cation contents, gas exchange rates and dry weights per unit length. These differences were eliminated by laboratory culture. Cultured shoots had increased affinity for both intra- and extracellular cadmium uptake compared to field-grown shoots. The results indicate that the activity of Cd and other divalent cations at the plasmalemma may control the affinity for and accumulation of Cd by this moss. The results are discussed in terms of the design of experiments in studies of ion uptake and tolerance in bryophytes.

Ionic control of intracellular and extracellular Cd uptake by the moss Rhytidiadelphus squarrosus (Hedw.) Warnst.

The effects of K, Ca, Mg and H ions on the kinetics of Cd uptake by the moss Rhytidiadelphus squarrosus (Hedw.) Warnst were investigated. The affinity of extracellular exchange sites for these ions decreased in the order Cd, H > Ca > Mg ≫ K and there were competitive interactions between ions. The affinity of the intracellular Cd transport site for these ions decreased in the order Ca > Cd > Mg ≫ K. Calcium was shown to be competitive and Mg a non-competitive inhibitor of intracellular Cd uptake. Intracellular Cd uptake, optimal at about pH 5.6, was highly sensitive to pH, which primarily affected the Vmax . Potassium, supplied at less than 1 mM, had no significant effect on Cd uptake. However, at concentrations greater than 1 mM, KNO3 caused both a stimulation in transport site activity and reduced affinity for Cd. Pretreatment with KNO3 removed potentially competing ions from the cell wall, altering the subsequent chemical equilibria established when moss shoots were incubated in Cd solutions. Reduced competition from ions removed by KNO3 pretreatment resulted in a higher affinity of both transport sites and extracellular exchange sites for Cd, Ca and Mg. The results demonstrate quantitatively the regulatory effects on intracellular metal uptake of the extracellular ionic environment provided by the cell wall.

Ring chromosome X in a child with manifestations of Kabuki syndrome.

A female patient with the karyotype 45,X/46, X, r(X)(p11.2 q13) and severe developmental delay, prominent fingertip pads, long palpebral fissures, short stature, and history of hypotonia had a phenotype reminiscent of Kabuki syndrome. We hypothesized that overexpression of X chromosome-derived sequences might be associated with the Kabuki-like phenotype observed. The nature and parental origin of this small-ring X were ascertained using a combination of genotyping with microsatellite markers and quantitative Southern blotting. PCR-based genotyping demonstrated heterozygosity at X-linked loci SBMA (Xq11-q12) and DXS227 (Xq13.1). Hemizygosity was observed at several loci: DMD STR-49 (Xp21.2), DXS1003 (Xp11.23), DXS988 (Xp11.21), DXS101 (Xq21.3), FMR-1 (Xq27.3), and DXYS64 (Xq28). This ring X chromosome is paternally derived since only maternal alleles are inherited at three informative microsatellite loci. Results of FISH and RT-PCR experiments indicate that the XIST locus is missing in the ring X chromosome and not expressed. These data indicated a large deletion of the X chromosome consistent with a small-ring X chromosome with approximate breakpoints near p11.2 and q13. These results are comparable to the observation of others where an atypically severe phenotype has been associated with the presence of an r(X), or small mar(X).

Community-acquired notifiable infectious diseases in a pediatric hospital: effect on infection control work load.

In a study of the impact of notifiable communicable disease admissions on infection control work load, we found notifiable disease cases to be responsible for less than 10% of admissions but 27% of infection control ward visits and to require extensive community liaison. Measures of work load based on nosocomial infections alone will underestimate infection control activity considerably.

Host resistance to mycobacteria is compromised by activation of the hypothalamic-pituitary-adrenal axis.

Host resistance to the growth of Mycobacterium avium and Mycobacterium tuberculosis is controlled by a gene, termed Nramp1, that maps to chromosome 1 in mice. Activation of the HPA axis or treatment of macrophages from susceptible mice with corticosterone suppresses the expression of Nramp1 mRNA and results in an increased susceptibility to mycobacterial growth. In contrast, neither activation of the HPA axis nor treatment of macrophages from resistant mice with corticosterone results in an alteration in their resistance or suppression of Nramp1 expression. Investigation into the mechanism of the differential response of the macrophages to corticosterone indicated that differences were associated with the stability of the mRNA in macrophages from BCG-resistant mice. Thus, corticosterone induced the accelerated degradation of Nramp1 mRNA as well as mRNA of several other macrophage activation genes in macrophages from BCG-susceptible mice. Treatment of macrophages with corticosterone before the induction of Nramp1 resulted in the accelerated degradation of mRNA in macrophages from both resistant and susceptible mice. The Nramp1 gene product appears to protect the mRNA of macrophage activation genes from degradation induced by corticosterone by an iron-dependent mechanism.

Interferons and bladder cancer.

With the introduction of BCG, intravesical instillation of immunotherapeutic agents has become a mainstay of therapy in the treatment of superficial bladder cancer. Interferon is capable of inducing a non-specific cellular and humoral immune response towards tumor cells. It has shown promise in reducing the recurrence and progression rates of superficial bladder cancer. In contrast to BCG, intravesical interferon is associated with minimal side effects and a very low dropout rate. Current research has focused on the use of interferon in combination with immunotherapeutic and cytotoxic drugs.

An evaluation of the use of electrophoresis and carbon furnace atomic absorption spectrometry to determine the copper level in separated serum protein fractions.

The results of an evaluation of the use of atomic absorption spectrometry with carbon furnace atomisation to analyse the copper content of individual protein fractions separated by electrophoresis is reported. Albumin copper levels in patients with rheumatoid arthritis were found to be about 1 to 2% of the total serum copper. Both albumin and alpha 2 levels are affected by contamination from residual copper present on the strip. An investigation of this contamination indicated very low blank copper levels in the areas of the strip containing alpha 1 and beta proteins. These results, and the results of analysis of copper in the ultrafiltrate from the same serum samples, are combined to define the copper distribution in a group of patients with rheumatoid arthritis. The determinations that can most readily be carried out by this technique are discussed.

Activity of the folate analog 10-ethyl, 10-deaza-aminopterin (10-EdAM) against human head and neck cancer xenografts.

In a previous study we established a markedly increased antitumour activity of 10-ethyl, 10-deaza-aminopterin (10-EdAM) as compared to methotrexate (MTX) when tested in vitro in squamous cell carcinoma cell lines from the head and neck (HNSCC). In this paper we describe the antitumour activity of these drugs in vivo in athymic nude mice bearing HNSCC xenografts. Using a schedule of 125 mg/kg i.p. for both drugs, injected on day 0 and 7, 10-EdAM caused a significant response in 2 out of 5 tumour lines, whereas MTX was completely inactive. These two lines moderately sensitive to 10-EdAM were not affected when the drug was given daily times 5 at an equitoxic dose of 0.75 mg/kg, indicating that the effect of the drug may be schedule dependent.

Resonance Raman spectroscopy of hemoglobin in intact cells: a probe of oxygen uptake by erythrocytes in rheumatoid arthritis.

Resonance Raman spectra from intact viable erythrocytes can be used to study oxygen uptake in solution. In addition to changes in the oxidation state marker (nu 4), other bands due to the porphyrin ring (nu 3) and vinyl modes indicate subtle changes at oxygen pressures close to where the T/R change occurs. A comparison of whole cell and lysate spectra indicates a partial denaturation of hemoglobin on lysis. A simple smear technique is used to measure spectra from rheumatoid and normal blood. Results indicate a faster but less complete uptake of oxygen in cells from patients with rheumatoid arthritis than is the case in normal cell populations.

The effect of seawater on the metabolism of some seashore and inland mosses.

Photosynthesis and protein synthesis were determined, by measuring uptake and incorporation of radioactive bicarbonate and L-valine, in some seashore and inland mosses after treatment with artificial seawater. In the inland species there was a progressive decline in both processes with increasing seawater concentration. Photosynthesis in the seashore mosses Grimmia maritima and Tortella flavovirens was unaffected by increasing seawater concentration, and supralittoral Ulota phyllantha was less seriously affected than epiphytic material from a subcoastal location. There was a marked decline in chlorophyll content and cessation of photosynthesis in the inland moss G. pulvinata after 3 days' seawater treatment whereas material treated with deionised water showed increased photosynthesis and chlorophyll levels over the same period. In G. maritima both chlorophyll levels and photosynthesis remained relatively unaffected by treatment with seawater for 4 days. Treatment of G. pulvinata with seawater and NaCl led to increased leakage of photosynthetic products to the external solution but this was insufficient to explain the reduction in carbon retained by the plant. The seawater-induced disruption of metabolism in inland bryophytes is believed to be primarily due to the uncontrolled entry of toxic ions into the moss cells whereas seawater tolerance in the seashore species probably reflects the possession of an efficient intracellular cation control mechanism.