CD36 polymorphisms and the age of disease onset in patients with pathogenic variants within the mutation cluster region of APC.

BACKGROUND: Familial adenomatous polyposis (FAP) is an autosomal dominant condition that predisposes patients to colorectal cancer. FAP is the result of a loss of APC function due to germline pathogenic variants disrupting gene expression. Genotype-phenotype correlations are described for FAP. For example attenuated forms of the disease are associated with pathogenic variants at the 5' and 3' ends of APC whilst severe forms of the disease appear to be linked to variants occurring in the mutation cluster region (MCR) of the gene. Variants occurring in the MCR are phenotypically associated with hundreds to thousands of adenomas carpeting the colon and rectum and patients harbouring changes in this region have a high propensity to develop colorectal cancer. Not all patients who carry pathogenic variants in this region have severe disease which may be a result of environmental factors. Alternatively, phenotypic variation observed in these patients could be due to modifier genes that either promote or inhibit disease expression. Mouse models of FAP have provided several plausible candidate modifier genes, but very few of these have survived scrutiny. One such genetic modifier that appears to be associated with disease expression is CD36. We previously reported a weak association between a polymorphism in CD36 and a later age of disease onset on a relatively small FAP patient cohort. METHODS: In the current study, we enlarged the FAP cohort. 395 patients all carrying pathogenic variants in APC were tested against three CD36 Single Nucleotide Polymorphisms (SNP)s (rs1049673, rs1761667 rs1984112), to determine if any of them were associated with differences in the age of disease expression. RESULTS: Overall, there appeared to be a statistically significant difference in the age of disease onset between carriers of the variant rs1984112 and wildtype. Furthermore, test equality of survivor functions for each SNP and mutation group suggested an interaction in the Log Rank, Wilcoxon, and Tarone-Ware methods for rs1049673, rs1761667, and rs1984112, thereby supporting the notion that CD36 modifies disease expression. CONCLUSIONS: This study supports and strengthens our previous findings concerning CD36 and an association with disease onset in FAP, AFAP and FAP-MCR affected individuals. Knowledge about the role CD36 in adenoma development may provide greater insight into the development of colorectal cancer.

Education of Sport University students regarding first-aid procedures after dental trauma.

AIM: The aim of this survey was to establish the current state of knowledge with regard to first-aid procedures and to compare the effectiveness of an educational lecture and a subsequent educational session. METHODS: A questionnaire to assess the attitudes and anticipated behaviours of Sport University students related to first-aid procedures following dental injury was administered to the students 3 times (after 3 and 12 months). A lecture on the subject of dental trauma was given just after the first questionnaire survey. A randomly selected students group received an extra educational task. RESULTS: The present study revealed a low level of initial knowledge of physical education students concerning first-aid measures in the case of dental trauma. A 30 minute lecture and an extra educational task significantly improved the knowledge evel. Even after one year the knowledge level was still high and sufficient to properly react when faced with dental trauma. CONCLUSION: Our research proves that the inclusion of dental trauma as a topic in the Sport University students' curricular training and paedagogical education should be introduced in the form of a clear and concise lecture.

Unexpected new roles for heparanase in Type 1 diabetes and immune gene regulation.

Heparanase (Hpse) is an endo-ß-d-glucuronidase that degrades the glycosaminoglycan heparan sulfate (HS) in basement membranes (BMs) to facilitate leukocyte migration into tissues. Heparanase activity also releases HS-bound growth factors from the extracellular matrix (ECM), a function that aids wound healing and angiogenesis. In disease states, the degradation of HS in BMs by heparanase is well recognized as an invasive property of metastatic cancer cells. Recent studies by our group, however, have identified unexpected new roles for heparanase and HS. First, we discovered that in Type 1 diabetes (T1D) (i) HS in the pancreatic islet BM acts as a barrier to invading cells and (ii) high levels of HS within the insulin-producing islet beta cells themselves are critical for beta cell survival, protecting the cells from free radical-mediated damage. Furthermore, catalytically active heparanase produced by autoreactive T cells and other insulitis mononuclear cells was shown to degrade intra-islet HS, increasing the susceptibility of islet beta cells to free radical damage and death. This totally novel molecular explanation for the onset of T1D diabetes opens up new therapeutic approaches for preventing disease progression. Indeed, administration of the heparanase inhibitor, PI-88, dramatically reduced T1D incidence in diabetes-prone NOD mice, preserved islet beta cell HS and reduced islet inflammation. Second, in parallel studies it has been shown that heparanase and HS can be transported to the nucleus of cells where they impact directly or indirectly on gene transcription. Based on ChIP-on-chip studies heparanase was found to interact with the promoters and transcribed regions of several hundred genes and micro-RNAs in activated Jurkat T cells and up-regulate transcription, with many of the target genes/micro-RNAs being involved in T cell differentiation. At the molecular level, nuclear heparanase appears to regulate histone 3 lysine 4 (H3K4) methylation by influencing the recruitment of demethylases to transcriptionally active genes. These studies have unveiled new functions for heparanase produced by T lymphocytes, with the enzyme mediating unexpected intracellular effects on T cell differentiation and insulin-producing beta cell survival in T cell-dependent autoimmune T1D.

Chemical and structural analysis of enhanced biochars: thermally treated mixtures of biochar, chicken litter, clay and minerals.

In this study biochar mixtures comprising a Jarrah-based biochar, chicken litter (CL), clay and other minerals were thermally treated, via torrefaction, at moderate temperatures (180 and 220 °C). The objectives of this treatment were to reduce N losses from CL during processing and to determine the effect of both the type of added clay and the torrefaction temperature on the structural and chemical properties of the final product, termed as an enhanced biochar (EB). Detailed characterisation indicated that the EBs contained high concentrations of plant available nutrients. Both the nutrient content and plant availability were affected by torrefaction temperature. The higher temperature (220 °C) promoted the greater decomposition of organic matter in the CL and dissociated labile carbon from the Jarrah-based biochar, which produced a higher concentration of dissolved organic carbon (DOC). This DOC may assist to solubilise mineral P, and may also react with both clay and minerals to block active sites for P adsorption. This subsequently resulted in higher concentrations of plant available P. Nitrogen loss was minimised, with up to 73% of the initial total N contained in the feedstock remaining in the final EB. However, N availability was affected by both torrefaction temperature and the nature of the clay minerals added.

Nonrandom DNA copy number changes related to lymph node metastases in squamous cell carcinoma of the lung.

Lung cancer is one of the most common malignancies and cancer-related death worldwide. Lymph node metastasis is the main cause of treatment failure. Although many studies were performed to evaluate genetic events associated with development and progression of lung cancer, molecular mechanism still remains poorly defined. In the present study, using comparative genomic hybridization (CGH) technique, we described the pattern of DNA copy number changes in a cohort of 42 primary squamous cell carcinomas (SCC) of the lung. A direct comparison of nonmetastatic (TxN0M0) and metastatic (TxN1-2M0) tumors was performed to define chromosomal imbalances related to lymph node metastases. Some genetic alterations were observed more frequently in metastatic than in non-metastatic tumors, including losses at 11q, 16p, 16q, 19p and gains at 4q, 7q, 12p, 13q, 18p. The gain at 7q with the smallest common altered region 7q31.2-q32, was found to be directly associated with lymph node involvement (p=0.0407). We suggest that the established chromosomal region harbors two putative tumor suppressor genes WNT2 and c-Met. An overexpresion of these genes seems to be involved in inducing the invasive growth and metastatic potential of SCC of the lung.

Molecular composition of the peri-islet basement membrane in NOD mice: a barrier against destructive insulitis.

AIMS/HYPOTHESIS: This study examined whether the capsule which encases islets of Langerhans in the NOD mouse pancreas represents a specialised extracellular matrix (ECM) or basement membrane that protects islets from autoimmune attack. METHODS: Immunofluorescence microscopy using a panel of antibodies to collagens type IV, laminins, nidogens and perlecan was performed to localise matrix components in NOD mouse pancreas before diabetes onset, at onset of diabetes and after clinical diabetes was established (2-8.5 weeks post-onset). RESULTS: Perlecan, a heparan sulphate proteoglycan that is characteristic of basement membranes and has not previously been investigated in islets, was localised in the peri-islet capsule and surrounding intra-islet capillaries. Other components present in the peri-islet capsule included laminin chains alpha2, beta1 and gamma1, collagen type IV alpha1 and alpha2, and nidogen 1 and 2. Collagen type IV alpha3-alpha6 were not detected. These findings confirm that the peri-islet capsule represents a specialised ECM or conventional basement membrane. The islet basement membrane was destroyed in islets where intra-islet infiltration of leucocytes marked the progression from non-destructive to destructive insulitis. No changes in basement membrane composition were observed before leucocyte infiltration. CONCLUSIONS/INTERPRETATION: These findings suggest that the islet basement membrane functions as a physical barrier to leucocyte migration into islets and that degradation of the islet basement membrane marks the onset of destructive autoimmune insulitis and diabetes development in NOD mice. The components of the islet basement membrane that we identified predict that specialised degradative enzymes are likely to function in autoimmune islet damage.

Structure and serological characterization of an Nepsilon-[(R)-1-carboxyethyl]-L-lysine-containing O-chain of the lipopolysaccharide of Proteus mirabilis O13.

In this paper we present the structure and describe serological properties of the O-specific polysaccharide of Proteus mirabilis O13 lipopolysaccharide, which contains a unique component: an amide of D-galacturonic acid (D-GalA) with an unusual amino acid, Nepsilon-[(R)-1-carboxyethyl]-L-lysine (alaninolysine, AlaLys). Selective chemical degradations of either GalA or AlaLys resulted in the loss of the serological reactivity of the polysaccharide with anti-O serum against P. mirabilis O13. Neither synthetic stereoisomers of AlaLys nor the isolated amide of GalA with AlaLys inhibited the reaction of the O-antiserum with the homologous lipopolysaccharide. The O-antiserum did not cross-react with the lipopolysaccharide of Providencia alcalifaciens O23 containing an amide of D-glucuronic acid with AlaLys. These data showed that both uronic acid and amino acid components of the amide play an important role in manifesting the P. mirabilis O13-specificity, but the full specific epitope also includes another O-specific polysaccharide component(s). A cross-reactivity of anti-O13 serum with some other P. mirabilis strains was observed and attributed to a common heat-stable antigen(s) different from the lipopolysaccharide.

The structure of the core region of the lipopolysaccharide from Klebsiella pneumoniae O3. 3-deoxy-alpha-D-manno-octulosonic acid (alpha-Kdo) residue in the outer part of the core, a common structural element of Klebsiella pneumoniae O1, O2, O3, O4, O5, O8, and O12 lipopolysaccharides.

The structure of lipid A-core region of the lipopolysaccharide (LPS) from Klebsiella pneumoniae serotype O3 was determined using NMR, MS and chemical analysis of the oligosaccharides, obtained by mild acid hydrolysis, alkaline deacylation, and deamination of the LPS: [carbohydrate structure see text] where P is H or alpha-Hep; J is H or beta-GalA; R is H or P (in the deacylated oligosaccharides). Screening of the LPS from K. pneumoniae O1, O2, O4, O5, O8, and O12 using deamination showed that they also contain alpha-Hep-(1-->4)-alpha-Kdo-(2-->6)-GlcN and alpha-Kdo-(2-->6)-GlcN fragments.

[Rare case of multiple primary malignant neoplasm of the larynx: squamous carcinoma and fibrosarcoma]. / Rzadki przypadek pierwotnego mnogiego nowotworu zlosliwego krtani: raka plaskonablonkowego i wlókniako-miesaka.

The problem of primary mixed neoplasms has been recently a subject of numerous research works trying to find out their pathogenetic mechanisms, early detection, clinical course and treatment opportunities. The authors present a rare case (fibrosarcoma and squamous carcinoma) in a 73-year-old man. The patient had a surgery (total laryngectomy) and complementary irradiation at the Institute of Oncology.

Bcl-2 antigen expression in patients with chronic hepatitis C and advanced fibrosis.

Bcl-2 antigen is a proto-oncogene responsible for the inhibition of the death of cells in the mechanism of apoptosis. The expression of bcl-2 antigen was analysed in liver bioptates obtained from patients with chronic hepatitis C. Thirty-five patients with advanced fibrosis (bridging fibrosis or cirrhosis) and various inflammatory activity were investigated. Control group was formed of the subjects with minimum or non-existent fibrosis. Histological and immunohistochemical assessments were performed with the use of widely-acceptable methods. The expression of bcl-2 antigen within portal spaces and lobules was evaluated. The values of lobular index declined as the fibrosis increased. As far as portal space index is concerned, no significant changes in correlation with fibrosis advancement were noticed. Additionally, the expression of bcl-2 antigen was recorded within lymphocyte infiltrations in lobules and portal spaces, in the epithelium of bile ducts in portal spaces and proliferating bile ductules in lobules as well as in perisinusoid cells. In hepatocytes, bcl-2 expression was detected more often among controls than in the analysed patients. The expression of bcl-2 antigen is directly proportional to the degree of inflammatory activity and it is inversely proportional to the degree of fibrosis. It seems that positive colour reaction with bcl-2 antigen in lobules may be of prognostic value for the prediction of collagen fibroplasia development. Bcl-2 expression in portal spaces has a limited diagnostic value for the prediction of the scope of fibrosis in patients with chronic hepatitis C.

Isolation using triflic acid solvolysis and identification of N(epsilon)-[(R)-1-carboxyethyl]-N(alpha)-(D-galacturonoyl)-L-lysine as a component of the O-specific polysaccharide of Proteus mirabilis O13.

An amino acid was released from the O-specific polysaccharide of Proteus mirabilis O13 by acid hydrolysis and identified as N(epsilon)-[(R)-1-carboxyethyl]-L-lysine by comparison with the authentic sample. An amide of this amino acid with D-galacturonic acid was isolated from the polysaccharide by solvolysis with anhydrous trifluoromethanesulfonic (triflic) acid and characterised by 1H and 13C NMR spectroscopy. These and published data enabled determination of the full structure of the repeating unit of the polysaccharide.

[Assessment of usefulness of PCNA and oncoprotein p53 staining in prediction of the recurrences in subjects operated on for laryngeal carcinoma]. / Ocena przydatnosci oznaczania atygenu proliferacyjnego PCNA i bialka p53 w przewidywaniu wznów u chorych operowanych z powodu raka krtani.

The authors estimated PCNA and P53 in subjects with laryngeal cancer with local or nodal recurrences. The study concerned 54 patients from Upper Silesia aged 37-79 (mean 57 +/- 8.8). The mean value of the PCNA index in subjects with local recurrence (LR) was 24.2% +/- 12.1 while in subjects without LR 22.1% +/- 9.4 (p > 0.05). Additionally, 21 subjects in whom no lymph node metastases were found during laryngectomy were separated from the investigated group. In 16 of them local recurrences were observed and the mean value of PCNA index was 30.9% +/- 12.5. In remaining 5 subjects in whom local recurrences were not developed the mean value of PCNA index was 21.7% +/- 11.2. The analysis of the P53 index in subjects with LR revealed significantly higher values (19.2% +/- 9.1) in comparison with cases without LR (13.2% +/- 6.3). The assessment of the mean values of PCNA and P53 index depending on T, N or stage as well as nodal recurrence did not reveal any statistical significance. Our study revealed usefulness of the P53 and PCNA as markers which could support the histological diagnostic process describing biology of the cancer cells. The demonstrated increase of PCNA and P53 index in patients with LR might be useful in prediction of LR.

The structure of the carbohydrate backbone of the core-lipid A region of the lipopolysaccharide from Proteus vulgaris serotype O25.

The following structure of the lipid A-core region of the lipopolysaccharide (LPS) from Proteus vulgaris serotype O25 was determined by using NMR and chemical analysis of the core oligosaccharide, obtained by mild acid hydrolysis of LPS, of the products of alkaline deacylation of the LPS, and of the products of LPS deamination: [structure: see text] Terminal residues of beta-GlcNAc and beta-Kdo (indicated by bold italics) are present alternatively in approximately 3:2 amount, leaving no unsubstituted beta-Gal. All sugars are in the pyranose form, alpha-Hep is the residue of L-glycero-alpha-D-manno-Hep, alpha-DDHep is the residue of D-glycero-alpha-D-manno-Hep.

Structural and serological studies of the related O-specific polysaccharides of Proteus vulgaris O21 and Proteus mirabilis O48 having oligosaccharide-phosphate repeating units.

The O-specific polysaccharide chains (O-antigens) of the lipopolysaccharides (LPSs) of Proteus mirabilis O48 and Proteus vulgaris O21 were found to have tetrasaccharide and pentasaccharide repeating units, respectively, interlinked by a glycosidic phosphate. Polysaccharides and an oligosaccharide were derived from the LPSs by various degradation procedures and studied by 1H and 13C NMR spectroscopy, including 2D COSY, TOCSY, NOESY, H-detected 1H,13C and 1H,31P HMQC experiments. The following related structures of the repeating units of the O-antigens were established (top: Proteus mirabilis O48; bottom: Proteus vulgaris O21) The O-specific polysaccharide of P. vulgaris O21 has the same structure as that of Hafnia allvei 744 and PCM 1194 [Petersson C., Jachymek, W., Klonowska, A., Lugowski, C., Niedziela, T. & Kenne, L. (1997) Eur. J. Biochem., 245, 668-675], except that the GlcN residue carries the N-acetyl rather than the N-[(R)-3-hydroxybutyryl] group. Serological investigations confirmed the close relatedness of the Proteus and Hafnia O-antigens studied.

[A case of laryngomucocele coexisted with pleomorphic adenoma of the larynx]. / Rzadki przypadek laryngomucocele z jednoczesnym wystapieniem gruczolaka wielopostaciowego krtani.

Benign pleomorphic adenomas of the larynx are very rare. Review of the literature showed about 20 cases of this tumours. We present a case of benign mixed tumor located in the false vocal cord and coexisted with laryngomucocele. The tumor was removed by endoscopic laser resection. To our knowledge it is the second published in literature case of benign plemorphic adenoma of the larynx in which treatment was performed by using CO2 laser surgery.

[Assessment of usefulness of PCNA and protein p53 in subjects with laryngeal cancer with local recurrence]. / Ocena przydatnosci oznaczania markera PCNA i bialka P53 u chorych na raka krtani ze wznowa miejscowa.

The authors estimated PCNA and P53 in subjects with laryngeal cancer in whom local or nodal recurrences were observed. The study included 54 patients from Upper Silesia in age 37-79 years (mean 57 +/- 8.8). The mean value of the PCNA index in subjects with local recurrence (LR) was 24.2% +/- 12.1 while in subjects without LR 22.1% +/- 9.4 (p > 0.05). Additionally, 21 subjects were separated from the investigated group in whom no lymph node metastases were found during laryngectomy. Among these subjects in 16 LR was observed (PCNA index was 30.9% +/- 12.5) while in remaining 5 subjects, in whom LR did not develop, PCNA index was 21.7% +/- 11.2. Analysis of the P53 index in subjects with LR revealed significantly higher values (19.2% +/- 9.1) in comparison to cases without LR (13.2% +/- 6.3). Our study revealed usefulness of the P53 and PCNA as markers which could support the histological diagnostic process describing biology of the cancer cells. The demonstrated increase of PCNA and P53 index in patients with LR might be useful in prediction of LR.

[The prospective analysis of laryngeal cancer growth dynamics in compliance with evaluation of DNA content and ploidy in cytologic material]. / Ocena prospektywna dynamiki rozrostu raka krtani z uwzglednieniem badania ploidii i zawartosci DNA w materiale cytologicznym.

The aim of this study was evaluation of DNA ploidy in the cytological samples from patients with laryngeal cancer and comparison a type of DNA ploidy with histological grading (G) and staging of neoplasmatic process. The examinations were performed on 47 patients operated due to laryngeal cancer (3 women, 44 men, mean age 58). Cytological material was collected from squamous carcinoma tissue by imprinting method. Slides were staining by Feulgen method in order to quantitative analysis of DNA by static cytophotometry. In studied material, aneuploid, polyploid and hypoploid tumors were found in 22 patients and diploid tumors were found in 25 cases. Analysis of DNA ploidy type distribution in correlation to G feature showed:--diploid type were dominated in G1 and G2 tumors,--aneuploid and polyploid type were dominated in G3 tumors. Hypoploid and polyploid tumors were appeared in patients with metastases to cervical lymph nodes more frequently than in patients without metastases. Analysis of DNA ploidy complete traditional histopathological diagnosis of laryngeal cancer and may be useful in predicting metastases to cervical lymph nodes.

[Assessment of utility of nm 23 antigen as a metastatic potential marker in laryngeal cancers: preliminary study]. / Ocena przydatnosci oznaczania antygenu nm 23 jako markera potencjalu przerzutowego raków krtani--doniesienie wstepne.

The aim of the study was the analysis of the utility of nm 23 protein in prediction of cervical lymph node metastases in patients with laryngeal cancer. A preliminary study was performed in 35 patients with laryngeal cancer with cervical lymph node metastases, which were confirmed by histopathologist. The control group consisted of 30 patients with laryngeal cancer without cervical lymph node metastases. In statistical analysis T and N were taken into account. In the investigated group with metastases the presence of positive immunostaining was found in 11% of cases while in the control group in 20%. The analysis of the presence of nm 23 protein revealed a weak usefulness of this marker as a factor which predicts the presence of the cervical metastases.

The effect of cell wall components on glycine-enhanced sterol side chain degradation to androstene derivatives by mycobacteria.

beta-Sitosterol side chain degradation by Mycobacterium sp. NRRL MB 3683 results in the formation of androstene derivatives and is increased in the presence of glycine. As the sterol transformation is carried out inside the cell, higher product accumulation could indicate faster diffusion of highly hydrophobic substrate through the cell wall permeability barrier. Cell wall preparations were obtained to analyse the effect of glycine on peptidoglycan components. Peptidoglycan is known to be the target for glycine action. In glycine-treated preparations, the molar ratio of diaminopimelic acid:muramic acid, the marker compounds of tetrapeptides and glycan strands respectively, was about 60% lower than in the control. This indicates a possible reduction in cross-linking between peptide units and the destruction of peptidoglycan. Unexpectedly, glycine also caused changes in the relative proportion of mycolic acids to other lipids occurring in the strain used for this study. The enhancement of beta-sitosterol side chain degradation is likely to result from disturbing the integrity of the cell wall components responsible for the permeability barrier in mycobacteria.

Clinical application of proliferating cell nuclear antigen, oncoprotein p53 and tumor front grading analysis in patients operated on for laryngeal cancer.

The authors assessed proliferating cell nuclear antigen (PCNA), p-53 oncoprotein and morphologic tumor front grading (TFG) in patients with advanced squamous cell carcinoma (SCC), of the larynx and a poor prognosis and tried to find a correlation with tumor stage, the Broders grading system, local and neck lymph node metastases, as well as nodal and local recurrences. In addition, utility of the parameters investigated was evaluated in developing a prognostic factor model, using uni- and multivariate Cox regression analysis. Included in this study were 54 patients (mean age 57 years +/- 8.6). PCNA-positive staining was found in all but one patient with advanced disease, while p-53 stained positively in only 24 subjects (44.4%). The PCNA index ranged from 4.6 to 59.0% (mean, 23.4 +/- 11. 0) and the p-53 index varied from 4.0 to 42.0% (mean, 17.2 +/- 8.6). The TFG score ranged from 9 to 23 points (mean, 15.1 +/- 3.2). PCNA, p-53 and TFG were found to be the markers that provided significant additional information about the biological behavior of tumor cells. The high variability of the results (PCNA, p-53) and high percentage of negatively stained cells (p-53) reduced their application in clinical use. PCNA correlated with tumor grade, G (r = 0.38; P < 0. 01), but negatively with nodal (N) disease(r = -0.37; P < 0.01). The mean values of PCNA and p-53 index were higher in the subgroup with local recurrences. Our present attempt to develop a useful prognostic factor model failed.