Genomewide search and association studies in a Finnish celiac disease population: Identification of a novel locus and replication of the HLA and CTLA4 loci

It has been reported that celiac disease (CD) is strongly associated with the HLA-DQ2 alleles DQA1*0501 and DQB1*0201. However, this association only accounts for a portion of the genetic component of CD. Several non-HLA loci and candidate genes that potentially contribute to CD susceptibility have been reported, but have not been confirmed. The aim of this study was to identify loci that contribute to disease susceptibility in a CD population from Finland. We performed a genomewide linkage scan and identified two regions of significant linkage to CD (6p and 2q23-32) and one region of suggestive linkage (10p). We also performed targeted typing and analyses that replicated the associations of the HLA and CTLA4 loci.     

Use of synthetic antigens to determine the epitope specificities of monoclonal antibodies against the 3-deoxy-D-manno-octulosonate region of bacterial lipopolysaccharide.

Mouse monoclonal antibodies were raised against heat-killed bacteria of the Re mutant R595 of Salmonella minnesota and characterized by the passive hemolysis and passive hemolysis inhibition tests and by double immunodiffusion experiments using lipopolysaccharide (LPS) from different rough mutants of S. minnesota and synthetic antigens. The latter were copolymerization products of acrylamide with the alpha- and beta-allylglycosides of 3-deoxy-D-manno-octulosonic acid (KDO) and the alpha-2,4-linked KDO disaccharide [poly-alpha-KDO, poly-beta-KDO, and poly-(alpha-KDO)2, respectively], and sodium (3-deoxy-D-manno-octulopyranosyl)onate-(2----6)-(2-deoxy-2-[ (R)-3- hydroxytetradecanoylamino]- beta-D-glucopyranosyl)-(1----6)-(2-deoxy-2-[(R)-3-hydroxytetradecanoy lam ino]-D-glucose) [alpha-KDO-(GlcNhm)2], representing a part structure of Re LPS. One antibody (clone 20, immunoglobulin M) was found to recognize a terminal alpha-linked KDO residue, since it reacted in the passive hemolysis assay with alpha-KDO-(GlcNhm)2 and all LPS tested, it was inhibited by all synthetic antigens containing alpha-linked KDO residues, and it gave a reaction of identity with poly-alpha-KDO and poly-(alpha-KDO)2 in double immunodiffusion experiments. A second antibody (clone 25, immunoglobulin G3) was identified as specific for an alpha-2,4-linked KDO disaccharide, since it reacted in immunodiffusion exclusively with synthetic poly-(alpha-KDO)2 and not with the monosaccharide derivatives in either anomeric configuration, and it was inhibited only with poly-(alpha-KDO)2 and with LPS from S. minnesota R595 (Re) and R345 (Rb2). The reaction of this antibody with R345 LPS is attributed to the quantitative substitution with KDO disaccharide present as a side chain, which is not present in stoichiometric amounts in the other LPS.     

Non-suppressed thyrotropin and elevated thyroglobulin are independent predictors of recurrence in differentiated thyroid carcinoma

OBJECTIVE: Although in most cases differentiated thyroid carcinoma (DTC) responds to surgery and radioiodine (RaI) therapy, some patients will have recurrence and eventually cancer-related death. However, although various prognostic factors of DTC have been identified (e.g. staging, suppressed thyrotropin), none of the previous studies have assessed simultaneously their role in multivariate analysis. DESIGN AND METHODS: In this retrospective population-based study, we reviewed the clinicopathological data of 254 DTC patients treated in eastern Finland during the years 1976-1995, for clinical characteristics, primary treatment, follow-up and cancer recurrence. Tumor stage was based on pathological tumor-node-metastasis (pTNM) classification, and histopathological specimens were re-evaluated. RESULTS: DTC recurrence occurred in 33 patients (13%). In univariate analyses, the predictors of recurrence were older age (>60 years, P<0.05), follicular tumor type (P<0.01), pTNM classification system (P<0.05) and post-ablative radioiodine uptake outside the neck (P<0.05). Non-suppressed serum thyrotropin (TSH) and elevated serum thyroglobulin (>3 microg/l) measured one year after operation were both related to tumor recurrence (P<0.05 and P<0.001 respectively). In multivariate analysis the independent predictors for recurrence were both elevated thyroglobulin (P<0.001) and non-suppressed TSH (P<0.05) independent of histology, pTNM stage and RaI uptake. Adjusted risk ratio for recurrence of DTC for unsuppressed thyrotropin was 2.3, for elevated thyroglobulin 14.0 and, if both conditions were present, the risk ratio increased to 45.1. CONCLUSION: Our results suggest that both non-suppressed serum TSH and elevated serum thyroglobulin are related to an increased risk of DTC recurrence independent of tumor type and pTNM stage.     

Production of extremely low volatile organic compounds from biogenic emissions: Measured yields and atmospheric implications

Oxidation products of monoterpenes and isoprene have a major influence on the global secondary organic aerosol (SOA) burden and the production of atmospheric nanoparticles and cloud condensation nuclei (CCN). Here, we investigate the formation of extremely low volatility organic compounds (ELVOC) from O₃ and OH radical oxidation of several monoterpenes and isoprene in a series of laboratory experiments. We show that ELVOC from all precursors are formed within the first minute after the initial attack of an oxidant. We demonstrate that under atmospherically relevant concentrations, species with an endocyclic double bond efficiently produce ELVOC from ozonolysis, whereas the yields from OH radical-initiated reactions are smaller. If the double bond is exocyclic or the compound itself is acyclic, ozonolysis produces less ELVOC and the role of the OH radical-initiated ELVOC formation is increased. Isoprene oxidation produces marginal quantities of ELVOC regardless of the oxidant. Implementing our laboratory findings into a global modeling framework shows that biogenic SOA formation in general, and ELVOC in particular, play crucial roles in atmospheric CCN production. Monoterpene oxidation products enhance atmospheric new particle formation and growth in most continental regions, thereby increasing CCN concentrations, especially at high values of cloud supersaturation. Isoprene-derived SOA tends to suppress atmospheric new particle formation, yet it assists the growth of sub-CCN-size primary particles to CCN. Taking into account compound specific monoterpene emissions has a moderate effect on the modeled global CCN budget.Formation and subsequent growth of new aerosol particles is a major source of cloud condensation nuclei (CCN) in the global troposphere (1, 2), and a big contributor to the large reported uncertainty in the radiative forcing by aerosol−cloud interactions (3–7). Multiple field studies have shown that CCN production is tightly connected with the oxidation of biogenic volatile organic compounds (BVOC) emitted by terrestrial ecosystems (8–11). To explain these observations, large-scale model simulations demonstrate a need for a BVOC oxidation mechanism in the atmosphere that produces very low volatility organic vapors with molar formation yields of at least a few percent per reacted precursor compound (12–14).The existence and formation mechanisms of essentially nonvolatile organic vapors in the atmosphere have puzzled scientists for some time (14–16). Such extremely low volatile organic compounds (ELVOC) (17) were recently detected, both in laboratory studies and in the ambient atmosphere (18), yet typical atmospheric oxidation chemistry schemes do not explain ELVOC produced on a time scale of minutes or hours. Furthermore, current state-of-the-art models using available chemistry schemes have systematically failed to reproduce the observed concentrations and volatility of organic aerosol components (17, 19). A plausible explanation for the fast ELVOC production was recently given by Ehn et al. (20), who proposed that highly oxidized (O:C ≈ 1) ELVOC are formed as first-generation oxidation products of α-pinene, a monoterpene emitted in vast quantities by different ecosystems. The authors proposed that α-pinene oxidation by ozone (O₃) forms peroxy radicals (RO₂), which undergo successive intramolecular hydrogen shifts followed by a rapid reaction with O₂, resulting in prompt production of high levels of oxygenation. This formation pathway, leading to highly oxygenated RO₂ radical and ELVOC formation, was more recently confirmed by Jokinen et al. (21) and Rissanen et al. (22). They showed that autoxidation, a mechanism known to be important in the liquid phase and which has been hypothesized to take place also in the gas phase (23), could mechanistically explain the gas-phase ELVOC formation. Their results also unambiguously demonstrated that highly oxidized RO₂ radicals and closed-shell ELVOC monomers (with a C₁₀ carbon skeleton) and dimers (C₂₀ carbon skeleton) are formed on time scales of seconds, thereby indicating immediate production of condensable vapors close to emission sources.Surprisingly, according to Ehn et al. (20), α-pinene appears to produce ELVOC with a much higher molar yield from ozonolysis (∼7%) than from the OH radical reaction (<1%). Jokinen et al. (21) investigated the relative importance of O₃ and OH radicals in the formation of highly oxygenated RO₂ from limonene and α-pinene, both having a reactive endocyclic double bond. They found that the amount of highly oxidized RO₂ radicals formed from the OH radical oxidation was only a small fraction of that from the pure ozonolysis reaction. The observed ELVOC yield from β-pinene ozonolysis in Ehn et al. (20) was much lower than that from α-pinene ozonolysis, indicating that the results obtained for α-pinene and limonene cannot be generalized to all BVOC, or even to all monoterpenes, present in the atmosphere. The relative yields of ELVOC and more volatile oxidized organics are key parameters for both regional and global CCN budgets, as their influence on atmospheric new particle formation as well as growth of both newly formed particles and sub-CCN-size primary particles can be markedly different (e.g., refs. 3 and 12).The mixture of BVOC emitted to the atmosphere contains a large number of compounds with different chemical structures (24). The magnitude and variability of these emissions are, however, not well understood. Even in the boreal forest, composed of only a few dominant tree species, BVOC emission patterns are complex, with prominent seasonal cycles (25) and large differences even between individual trees of the same species (26). In the tropics, with more diverse vegetation properties, relatively few BVOC emission assessments have been published so far (e.g., refs. 27 and 28). Future BVOC emission characteristics are expected to change considerably in many regions as a result of increasing temperatures and CO₂ concentrations, as well as changes in extreme weather conditions and vegetation cover (25, 29–31).In this work, we investigate ELVOC formation and associated CCN production from BVOC species having different chemical structures. We determine ELVOC yields from both ozonolysis and OH radical-initiated oxidation of five major BVOC species through comprehensive laboratory experiments. These species include monoterpenes with endocyclic and exocyclic double bonds and acyclic compounds with reactive double bonds (hereafter referred as endocyclic, exocyclic, and acyclic terpenes) as well as isoprene. Based on the observed ELVOC yields, we then use global model simulations to investigate the implications of our findings on atmospheric CCN production.     

Nitric oxide and proinflammatory cytokines in nasal lavage fluid associated with symptoms and exposure to moldy building microbes

Epidemiological data indicate that living or working in a moldy building is associated with increased risk of respiratory symptoms and disease related to inflammatory reactions, but biochemical evidence linking cause and effect is still scarce. The staff working in a mold-contaminated school, and a reference group without such exposure, were studied. Nasal lavage was performed and health data were collected with a questionnaire at the end of the spring term, after a 2.5-mo summer vacation, and at the end of the fall term. Here we show that concentrations of tumor necrosis factor alpha (TNF-alpha), interleukin-6 (IL-6), and nitric oxide (NO) in nasal lavage fluid were significantly higher in the exposed than in the control subjects at the end of the first exposure period. These inflammatory mediators decreased to reference group concentrations during the period when there was no exposure and the production of NO and IL-6 increased again during the reexposure in the fall term. Reports of cough, phlegm, rhinitis, eye irritation, and fatigue paralleled the changes in the measured inflammatory markers. These results point to an association between inflammatory markers in the nasal lavage fluid, the high prevalence of respiratory symptoms among the occupants, and chronic exposure to molds in the indoor environment.     

Hypofractionated radiotherapy with concurrent 5-fluorouracil radiosensitisation for recurrent or locally advanced colorectal cancer A phase II study

In a pilot study we treated 19 patients suffering from recurrent or locally advanced inoperable colorectal cancer, with concurrent hypofractionated radiotherapy (4 – 5 Gy/fraction, 2 fractions per week) and 5-Fluorouracil bolus, 1 hour before RT at doses of 300 mg/m². For 16 patients treated with radical intent the Normalised Total Dose for α/β = 10 Gy ranged between 56 – 74 Gy (median 62 Gy). The schedule used was very well tolerated. Moderate grade II haematological toxicity was observed in 11% of cases and diarrhoea grade II/III resulting in 2 – 4 weeks treatment delay was observed in 26% of cases. One case with bowel perforation and one with painful subcutaneous fibrosis was observed during 12 – 27 months of follow-up. Out of 16 patients treated with radical intent 4 (25%) showed complete response and the overall response rate was 56% (9/16). The one year symptom free survival was obtained in 11/16 (69%) radically treated patients. It is concluded that hypofractionated radiochemotherapy with 5-Fluorouracil for recurrent or locally advanced colorectal cancer is an effective regimen and has acceptable acute and late toxicity. Further investigation is required.

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Résumé. Dans un projet pilote, nous avons traité 19 patients souffrant d'un cancer colo-rectal récidivant ou localement avancé inopérable par l'administration simultanée d'une radiothérapie hypofractionnée (4 – 5 Gy/fraction à raison de 2 fractions par semaine) et bolus de 5-Fluorouracil à la dose de 300 mg/m² une heure avant la radiothérapie. Chez 16 patients traités dans un but radical, la dose totale normalisée a été de 56 – 74 Gy (moyenne 62 Gy). Le schéma utilisé a été particulièrement bien toléré. Une toxicité hématologique modérée de grade II a été observée chez 11% des patients et une diarrhée de grade II/III entra?nant un délai dans le traitement de 2 à 4 semaines a été observée chez 26% des patients. Une perforation intestinale et une fibrose sous-cutanée douloureuse ont été observées au cours de la période postopératoire de 12 à 27 mois. Quatre des 16 patients traités à des fins curatives (25%) ont montré une résponse complète et une réponse globale a été observée chez 56% des patients (9/16). Une survie sans sympt?mes à un an a été obtenue chez 11 des 16 patients (69%). On conclut que l'assocation de 5-Fluorouracil et d'une radiothérapie hypofractionnée constitue un schéma thérapeutique effectif en cas de cancer colorectal récidivant ou localement avancé avec une toxicité aigu? et tardive acceptable. Des investigations complémentaires sont nécessaires.

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Accepted: 20 June 1996     

Regional lymph node reactions related to hormone receptor status in female breast carcinoma

A series of 211 female breast carcinomas was studied with special reference to the relationships between estrogen (ER) and progesterone (PR) receptor status and the reactions of the axillary lymph nodes as manifestations of tumor-host reactivity. Carcinomas were classified according to their nuclear grade (NG), and the following nodal reactions were recorded: extent of cancer metastasis, paracortical activity (PCA), follicular hyperplasia (LFH), sinus histiocytosis (SH), and degenerative sinus histiocytosis (DSH). NG did not directly correlate with ER- or PR-positivity, albeit the highest frequency of both was found in well differentiated carcinomas. ER- and less significantly PR-values were higher in post- than in premenopausal females. ER- and PR-positivity as well as the absolute receptor values were lowest in the cases with extensive (Grade 4) nodal metastases, but did not show a direct relationship to the other grades (0 to 3). PCA did not show any relation to the receptor status, whereas LFH seemed to bear an inverse relationship with ER- (but not PR-) positivity. SH seemed to be associated with higher frequency of ER-positivity than did DSH-nodes, the differences being slight, however. It is concluded that hormone receptor assays connected with the morphological assessment of tumor-host reactivity provide some benefit in grouping the breast cancer patients according to the different expectancy of the outcome of their disease.     

Clinical relevance of p53 index and expression of proliferating cell nuclear antigen and Ki-67 in gastric cancer

The prognostic value of the immunohistochemical expression of p53 protein, proliferating-cell nuclear antigen (PCNA) and Ki-67 antigen was evaluated in a series of 116 stage I–II gastric cancer patients. The staining for p53 protein (staining frequency and intensity) in malignant cells was expressed as a p53 index. Similarly, the staining frequency and intensity for PCNA and Ki-67 were evaluated. The p53 index was independent of the stage and differentiation grade, but significantly related to DNA ploidy, S-phase fraction and mitotic activity. A high p53 index was a sign of inferior survival, compared to a low or intermediate index. p53-negative tumours were also associated with poor survival. In a multivariate analysis, only the depth of tumour infiltration and the presence of nodal metastases were independent prognostic factors in stage I–II gastric cancer. PCNA expression and Ki-67 antigen expression were not related to the stage, ploidy, proliferative activity or p53 expression, and they had no impact on survival. The results indicate that p53 protein expression may be of prognostic significance in gastric cancer, while PCNA and Ki-67 antigen expression have no predictive value. Received: 2 February 1998 / Accepted: 16 June 1998     

Synthesis and immunochemical characterization of neoglycoproteins containing epitopes of the inner core region of Pseudomonas aeruginosa RNA group I lipopolysaccharide

The disaccharide allyl 2-acetamido-2-deoxy-alpha-D-galactopyranosyl-(1-->3)-7-O-carbamoyl-L-glycero-alpha-D-manno-heptopyranoside 5 (GalNAc-cmHep-allyl) was synthesized starting from 1 and 2. Compound 5, cmHep-allyl and the disaccharide cmHep-(1-->3)-Hep-allyl were converted into cysteamine-spacered derivatives and conjugated to bovine serum albumin (BSA) to yield the neoglycoconjugates 7--9, respectively. These conjugates were used to immunize mice and to prepare monoclonal antibodies (mAbs) which were characterized in comparison to mAbs obtained after immunization with heat-killed Pseudomonas aeruginosa strain H4. Two antibodies obtained after immunization with the neoglycoconjugates bound strongly to cmHep-BSA and with lower affinity to cmHep-Hep-BSA but did not bind to GalNAc-cmHep-BSA or to H4 LPS. Another antibody obtained after immunization with heat-killed bacteria bound to LPS and GalNAc-cmHep-BSA but not to cmHep-Hep-BSA or cmHep-BSA     

Matrix metalloproteinase-19 expression in dermal wounds and by fibroblasts in culture

Here, we have examined the expression of matrix metalloproteinase-19 (MMP-19) in human cutaneous wounds and by human skin fibroblasts in culture. Expression of MMP-19 was detected by immunohistochemistry in fibroblasts, capillary endothelial cells, and macrophages in the dermal layer of large granulating wounds, as well as in chronic venous and decubitus ulcers. MMP-19 mRNA expression and pro-MMP-19 production by dermal fibroblasts in culture was potently enhanced by tumor necrosis factor-alpha (TNF-alpha). Induction of MMP-19 expression by TNF-alpha was prevented partially by blocking the activation of extracellular signal-regulated kinase (ERK)-1/2 by PD98059 and p38 activity by SB203580. Activation of ERK1/2 by adenovirus-mediated delivery of constitutively active MAPK/ERK kinase 1 resulted in the induction of MMP-19 expression. Activation of p38 alone by adenovirally delivered constitutively active MAPK kinase 3b (MKK3b) and MKK6b also enhanced MMP-19 production, and the most potent induction of MMP-19 expression was noted when ERK1/2 was activated in combination with p38. Activation of c-Jun N-terminal kinase (NK). Abundant pro-MMP-19 production by fibroblasts was associated with proteolytic processing of secreted pro-MMP-19. These results suggest a role of MMP-19 in cutaneous wound repair and identify three distinct signaling pathways, which coordinately mediate induction of MMP-19 expression in fibroblasts: mitogen-activated ERK1/2 pathway and stress-activated JNK and p38 pathways, of which control proteolytic activity of dermal fibroblasts.