Tomasz Olszowski

Co-Complexes of MASP-1 and MASP-2 Associated with the Soluble Pattern-Recognition Molecules Drive Lectin Pathway Activation in a Manner Inhibitable by MAp44

The lectin pathway of complement is an integral component of innate immunity. It is activated upon binding of mannan-binding lectin (MBL) or ficolins (H-, L-, and M-ficolin) to suitable ligand patterns on microorganisms. MBL and ficolins are polydisperse homo-oligomeric molecules, found in complexes with MBL-associated serine proteases (MASP-1, -2, and -3) and MBL-associated proteins (MAp19 and MAp44). This scenario is far more complex than the well-defined activation complex of the classical pathway, C1qC1r2C1s2, and the composition of the activating complexes of the lectin pathway is ill defined. We and other investigators recently demonstrated that both MASP-1 and MASP-2 are crucial to lectin pathway activation. MASP-1 transactivates MASP-2 and, although MASP-1 also cleaves C2, MASP-2 cleaves both C4 and C2, allowing formation of the C3 convertase, C4bC2a. Juxtaposition of MASP-1 and MASP-2 during activation must be required for transactivation. We previously presented a possible scenario, which parallels that of the classical pathway, in which MASP-1 and MASP-2 are found together in the same MBL or ficolin complex. In this study, we demonstrate that, although MASPs do not directly form heterodimers, the addition of MBL or ficolins allows the formation of MASP-1–MASP-2 co-complexes. We find that such co-complexes have a functional role in activating complement and are present in serum at varying levels, impacting on the degree of complement activation. This raises the novel possibility that MAp44 may inhibit complement, not simply by brute force displacement of MASP-2 from MBL or ficolins, but by disruption of co-complexes, hence impairing transactivation. We present support for this contention.

The Effects of Cadmium at Low Environmental Concentrations on THP-1 Macrophage Apoptosis.

Cadmium at environmental concentrations is a risk factor for many diseases, including cardiovascular and neurodegenerative diseases, in which macrophages play an important role. The aim of this study was to evaluate the effects of cadmium at low environmental (nanomolar) concentrations on apoptotic processes in THP-1(acute monocytic leukemia cells line)-derived macrophages, with special focus on mitochondrial events involved. Macrophages were incubated with various cadmium chloride (CdCl2) solutions for 48 h at final concentrations of 5 nM, 20 nM, 200 nM and 2 µM CdCl2. Cell viability was measured using flow cytometry. Flow cytometric measurement (annexin V/FITC (annexin V/fluorescein isothiocyanate) and PI (propidium iodide) double staining) was used to quantify the extent of apoptosis. Fluorescence and confocal microscopy were used for imaging of apoptosis process. Changes in mitochondrial membrane potential were monitored using cytofluorimetry after cell staining with JC-1(5,5′,6,6′-tetrachloro-1,1′,3,3′-tetraethylbenzimidazol-carbocyane iodide) probe. Mitochondrial ROS (reactive oxygen species) levels were measured cytofluorimetrically after incubation of cells with mitochondrial superoxide indicator (MitoSOX) red fluorescent marker. The mRNA expression of Bcl-2 and Bax was analysed with qRT-PCR. Our study demonstrates that cadmium, even at low environmental concentrations, exerts mitochondrial toxicity in THP-1 macrophages. Forty-eight-hour exposure to very low concentrations reduces cell viability and results in cell death by apoptosis and necrosis. The decrease in mitochondrial membrane potential, increased ROS production, increased Bax and decreased Bcl-2 mRNA expression are mitochondrial events involved in cadmium-induced apoptosis.

The use of the transparotid approach for surgical treatment of condylar fractures – Own experience

AIM To assess the results of surgical treatment of condylar fractures using the transparotid approach. MATERIAL AND METHODS The transparotid approach was used in 38 patients with unilateral condylar fracture. In four cases a single 2.0 plate was used, in the latter - 3D plates were used. All the patients were subjected to control clinical examination including: occlusion, facial nerve function, mandibular movements, pain presence, subjective assessment of the scar aesthetics and the presence of salivary fistula or salivary cyst on the first day following surgery and after 1, 3 and 6 months. Control radiography of the mandible in at least two projections was made on the first day after surgery and after 3 months. RESULTS In 3 patients a partial paresis of the facial nerve was noticed followed by a spontaneous recovery 3 months postoperatively. In 2 patients acoustic effects, without pain in the temporomandibular joint of the fractured side were still present 6 months postoperatively. Plate fractures were found in two out of four patients operated on with single-plate technique. Loosening and displacement of a fixation screw occurred in 4 patients; in 3 cases it referred to a single 2.0 plate and in one, a Delta plate. Post-operation scar was accepted by all the patients. CONCLUSION The transparotid approach allows for direct visualisation of the fracture providing proper reduction and osteosynthesis, with a low risk of facial nerve paresis. Precise wound closure in layers, especially of the parotid capsule allows avoiding a salivary fistula.

Lectin pathway of complement activation in a Polish woman with MASP-2 deficiency

Functional MASP-2 deficiency, considered a primary immunoeficiency (PID), is caused in Caucasians by homozygosity for the ASP-2 D120G mutation. It was estimated that homozygosity of his mutation occurs in 6 out of 104 individuals (García-Laorden t al., 2006, 2008; Stengaard-Pedersen et al., 2003). Functional ASP-2 deficiency in Caucasians is only found in such individals where the MASP-2 (generally at low levels) is incapable of ssociating with the pattern recognition molecules, MBL and ficolns (Stengaard-Pedersen et al., 2003). To our knowledge, only nine ases of this disorder of complement system have been reported o far (Cedzynski et al., 2004; García-Laorden et al., 2006, 2008; lesen et al., 2006; Sørensen et al., 2005; Stengaard-Pedersen et al., 003; Stover et al., 2005), of them the case described by Stengardedersen et al. being the best characterized (Stengaard-Pedersen t al., 2003). Most of these cases (5 cases) did not present with any ymptoms, suggesting incomplete clinical penetrance of MASP-2 eficiency (García-Laorden et al., 2006, 2008; Stover et al., 2005). he impact of MASP-2 deficiency on disease susceptibility remains nresolved. We present a 19-year-old Polish Caucasian woman, homozyous for MASP2 D120G mutation, in whom the mutation was found y chance while conducting a case–control study on possible assoiation of MBL2 and MASP2 gene polymorphisms with dental caries Olszowski et al., 2012). The protocol of the new case study was submitted to and pproved by the Bioethical Committee of the Pomeranian Medical niversity (decision reference number KB-0012/130/12). Written ormal consent for blood sample collection, genetic and biochemcal analyses of components of lectin pathway was obtained from he patient. The physical examination and consultation with the patient’s amily physician revealed that the case patient is essentially ealthy. Apart from the dry skin and features of allergic eczema o cobalt (low-grade) no deviations were found in the patient. ll vaccinations so far well tolerated, with no wrong reacions. A retrospective review of the patient’s chart revealed, at he age of 13 years, a few episodes (5–6) of syncope without bvious cause that disappeared after 2 years. In the patient’s istory no recurrent infections or autoimmune disorders were eported.

The Effect of Cadmium on COX-1 and COX-2 Gene, Protein Expression, and Enzymatic Activity in THP-1 Macrophages

The aim of this study was to examine the effects of cadmium in concentrations relevant to those detected in human serum on cyclooxygenase-1 (COX-1) and cyclooxygenase-2 (COX-2) expression at mRNA, protein, and enzyme activity levels in THP-1 macrophages. Macrophages were incubated with various cadmium chloride (CdCl2) solutions for 48 h at final concentrations of 5 nM, 20 nM, 200 nM, and 2 μM CdCl2. The mRNA expression and protein levels of COXs were analyzed with RT-PCR and Western blotting, respectively. Prostaglandin E2 (PGE2) and stable metabolite of thromboxane B2 (TXB2) concentrations in culture media were determined using ELISA method. Our study demonstrates that cadmium at the highest tested concentrations modulates COX-1 and COX-2 at mRNA level in THP-1 macrophages; however, the lower tested cadmium concentrations appear to inhibit COX-1 protein expression. PGE2 and TXB2 production is not altered by all tested Cd concentrations; however, the significant stimulation of PGE2 and TXB2 production is observed when macrophages are exposed to both cadmium and COX-2 selective inhibitor, NS-398. The stimulatory effect of cadmium on COXs at mRNA level is not reflected at protein and enzymatic activity levels, suggesting the existence of some posttranscriptional, translational, and posttranslational events that result in silencing of those genes’ expression.

DD Genotype of ACE I/D Polymorphism Might Confer Protection against Dental Caries in Polish Children

The aim of the study was to examine the frequencies of the genotypes and alleles of ACE insertion/deletion (I/D) polymorphism and their association with dental caries in a sample of Polish children. The study subjects were 120 children with dental caries experience (cases) and 41 caries-free individuals (controls). The genotyping was performed using polymerase chain reaction. The genotype distributions of ACE I/D polymorphism were not statistically different between carious and control children. However, we found a borderline overrepresentation of the II + ID genotypes versus the DD genotype in the carious compared to the control group (69.2% and 51.2%, respectively, p = 0.057). Logistic regression analysis adjusted for age and sex revealed that I allele carriage was a significant predictor of dental caries susceptibility (OR = 2.14, 95% CI = 1.02-4.49, p = 0.041). In conclusion, the DD genotype of ACE I/D polymorphism might be protective against dental caries in Polish children.

The Lack of Association between FCN2 Gene Promoter Region Polymorphisms and Dental Caries in Polish Children

The aim of this study was to examine the association of single-nucleotide polymorphisms (SNPs) in the gene encoding ficolin-2 protein (FCN2 gene) at positions -986 (rs17514136), -602 (rs3124953), and -4 (rs3124952) with dental caries in Polish children. Two hundred and sixty Polish Caucasian children aged 15 years were enrolled in this study: 82 with “higher” caries experience (DMFT >5) and 178 with “lower” caries experience (DMFT ≤5). In addition, subjects with caries experience (DMFT ≥1) and caries-free subjects (DMFT = 0) were compared. FCN2 SNPs were genotyped with PCR-RFLP methods. There were no significant differences in the genotype, allele, or haplotype distributions in 3 analyzed SNPs of the FCN2 gene between children with “higher” and those with “lower” caries experience as well as between children with caries experience and caries-free children. In conclusion, we did not find any association of FCN2 promoter polymorphisms at positions -986, -602, and -4 with dental caries in Polish children.

The influence of the place of residence, smoking and alcohol consumption on bone mineral content in the facial skeleton

BACKGROUND Environmental factors exert their influence on the living organism throughout ontogeny. More and more often, researchers find correlations between specific environmental factors and the so-called diseases of affluence. Deficits and excess of essential elements also leave their mark on the skeleton. AIM To investigate the influence of alcohol consumption, tobacco smoking and place of residence, according to sex and calendar age, on the concentrations of micro-, macro- and toxic elements in human facial bones. MATERIAL & METHODS Patients undergoing surgical treatment were examined for the mineral content in the collected bone material. The bone contents of the following elements were determined: Ca, K, Mg, Na, P, Fe, Zn, Mo, Ba, Mn, Li, Be, Co, B, Sr, Cr, Pb, Cd, Ni, and Al, depending on the type of facial bone, sex, calendar age, alcohol consumption, tobacco smoking and place of residence. RESULTS Sex and alcohol consumption showed the highest degree of correlation with the content of the minerals included in the study. Alcohol drinking was found to exert the strongest influence on womens bodies, the highest number of statistically significant correlations was demonstrated between the content of minerals in the examined bones and alcohol drinking in women. Other factors included in the analysis had a different impact on men and women, the concentrations of elements included in the study differed depending on age, tobacco smoking and place of residence. CONCLUSIONS The observed differences in the element mineral composition of the human facial skeleton may be explained by developmental specifics and functional adaptation. However, general biological characteristics (sex, age), environmental factors (place of residence), as well as smoking and alcohol use may exert significant influence on the concentrations of micro-, macro- and toxic elements in particular regions of the human skeleton. The impact of environmental factors is a very complex phenomenon, which may be stronger or more subtle, leaving its mark on the bone structure. The environmental factors included in the analysis had a different influence on men than women.

The Yngve Ericsson Prize in Preventive Odontology

Patent Revenue Fund and ORCA herby solicit nominations for the 2016 Yngve Ericsson Prize. The award will be SEK 300,000 (approx. EUR 32,000) for one person and SEK 200,000 per person if shared by two recipients. The Prize winner(s) will be selected by a Prize Committee of distinguished scientists, three members of which are appointed by the Patent Revenue Fund for Preventive Odontology and two members appointed by ORCA. The Prize is awarded to persons who have performed outstanding laboratory or clinical research that has contributed specifically to the prevention of dental and oral disease. Candidates are judged on the originality, quality and range of their scientific contributions as well as the range and clinical importance of the results. Individuals who are still active in research are preferred candidates. No preference will be given to candidates from any country. A nomination should contain the name of the candidate and must be accompanied by a statement giving reasons why the candidate would be a worthy awardee, including a list of the candidate’s relevant scientific publications and, if possible, a short CV. Nominations of candidates must be received by the Secretary of the Patent Revenue Fund, Prof. Peter Lingström (Institute of Odontology, Box 450, SE–405 30 Göteborg, Sweden; E-Mail peter.lingström @ odontologi. gu.se), no later than February 15, 2016. Call for Nominations

Nieswoiste czynniki odpornościowe w przebiegu choroby próchnicowej

Streszczenie Prochnica zebow nalezy do najbardziej rozpowszechnionych chorob na świecie. Dzialanie nieswoistych czynnikow odpornościowych zawartych w ślinie jest ukierunkowane na zapobieganie nadmiernemu namnazaniu sie drobnoustrojow, a nie na eliminacje określonych ich gatunkow. Nieswoiste czynniki odpornościowe zawarte w ślinie, jak mucyny, bialka bogate w proline, immunoglobuliny, aglutynina, laktoferryna, cistatyny i lizozym, a takze defensyny, katelicydyna i histatyny stanowią pierwszą linie ukladu odpornościowego jamy ustnej. Ich wzajemne interakcje i wspoldzialanie z czynnikami swoistymi nie zostaly dokladnie poznane, z calą pewnością jednak nie udalo sie stwierdzic korelacji pomiedzy stezeniem ktoregokolwiek pojedynczego niespecyficznego czynnika obronnego w ślinie a podatnością na prochnice. Nalezy jednak pamietac, ze wartośc wskaźnika PUW stanowi wynik kumulacji skutkow choroby prochnicowej przez cale dotychczasowe zycie i niekoniecznie odzwierciedla jej aktywnośc w danym momencie.