Atypical Hemolytic Uremic Syndrome (aHUS) and Adenosine Deaminase (ADA)-Deficient Severe Combined Immunodeficiency (SCID)-Two Diseases That Exacerbate Each Other: Case Report.

Hemolytic uremic syndrome (HUS) is defined by the triad of microangiopathic hemolytic anemia, thrombocytopenia, and acute kidney injury (AKI). Atypical HUS (aHUS), distinguished by its etiology, is caused by uncontrolled overactivation of the alternative complement pathway. The correct diagnosis of aHUS is complex and involves various gene mutations. Severe combined immunodeficiency (SCID), characterized by severe T-cell lymphocytopenia and a lack of antigen-specific T-cell and B-cell immune responses, is of seldom occurrence. In 10-15% of pediatric patients, SCID is caused by adenosine deaminase (ADA) deficiency. The authors describe the case of a boy who suffered from both aHUS and ADA-deficient SCID. At the age of 9 months, the patient presented acute kidney injury with anuria and coagulopathy. The diagnosis of aHUS was established on the basis of alternative complement pathway deregulation and disease-associated gene mutations. Further examination revealed immune system failure and, at the age of 13 months, the ADA deficiency was confirmed by genetic tests and the boy was diagnosed with ADA-SCID. ADA SCID has recently been described as a possible triggering factor of aHUS development and progression. However, more research is required in this field. Nevertheless, it is crucial in clinical practice to be aware of these two co-existing life-threatening diseases.

Biological Activity of Naturally Derived Naphthyridines.

Marine and terrestrial environments are rich sources of various bioactive substances, which have been used by humans since prehistoric times. Nowadays, due to advances in chemical sciences, new substances are still discovered, and their chemical structures and biological properties are constantly explored. Drugs obtained from natural sources are used commonly in medicine, particularly in cancer and infectious diseases treatment. Naphthyridines, isolated mainly from marine organisms and terrestrial plants, represent prominent examples of naturally derived agents. They are a class of heterocyclic compounds containing a fused system of two pyridine rings, possessing six isomers depending on the nitrogen atom's location. In this review, biological activity of naphthyridines obtained from various natural sources was summarized. According to previous studies, the naphthyridine alkaloids displayed multiple activities, i.a., antiinfectious, anticancer, neurological, psychotropic, affecting cardiovascular system, and immune response. Their wide range of activity makes them a fascinating object of research with prospects for use in therapeutic purposes.

Genetic background of atypical hemolytic-uremic syndrome and its influence on the course of disease and therapeutic effects

Atypical hemolytic-uremic syndrome (aHUS) results from excessive, uncontrolled activation of the alternative pathway of the complement system. It is important to distinguish aHUS from other thrombotic microangiopathies. The aim of this paper is to discuss the complexity and relevance of the genetic background of aHUS patients. The review discusses the genetic variants that are important for diagnosis, treatment and prognosis of patients, which is inevitably important for the qualification of patients for treatment with eculizumab. These variants are not only found in the genes involved in the control of complement system but are also related to the coagulation system. The paper emphasizes the diagnostic difficulties resulting from the extremely diverse genetic background of the patients. It is important to conduct further genetic studies of aHUS patients, also paying attention to genes unrelated to the complement system. The paper contains information on the role of genetic predisposition in tailoring the risk for aHUS and determining its clinical outcome, including qualification for eculizumab therapy.

The rs2910164 Genetic Variant of miR-146a-3p Is Associated with Increased Overall Mortality in Patients with Follicular Variant Papillary Thyroid Carcinoma.

Aberrant expression of the sodium-iodide symporter (NIS) and the resistance to post-operative radioactive iodide treatment is a crucial cause of higher mortality of some thyroid cancer patients. In this study, we analyzed the impact of miR-146a on the expression and function of NIS and on the overall survival of thyroid cancer patients. The study included 2441 patients (2163 women; 278 men); including 359 cases with follicular variant of papillary thyroid carcinoma (fvPTC). miR:NIS interactions were analyzed in cell lines using in vivo binding and inhibition assays and radioactive iodine uptake assays. Tumor/blood DNA was used for rs2910164 genotyping. Overall survival was assessed retrospectively. In the results, we showed that miR-146a-3p directly binds to and inhibits NIS. Inhibition of miR-146a-3p restores the expression and function of NIS, increasing radioactive iodine uptake. Rs2910164 functional variant within miR-146a-3p is associated with increased overall mortality among fvPTC female patients. The deaths per 1000 person-years were 29.7 in CC carriers vs. 5.08 in GG/GC-carriers (HR = 6.21, p = 0.006). Higher mortality of CC vs. GG/GC carriers was also observed in patients with lower clinical stage (HR = 22.72, p < 0.001), smaller tumor size (pT1/pT2) (HR = 25.05, p < 0.001), lack of extrathyroidal invasion (HR = 9.03, p = 0.02), lack of nodular invasion (HR = 7.84, p = 0.002), lack of metastases (HR = 6.5, p = 0.005) and older (age at diagnosis >50 years) (HR = 7.8, p = 0.002). MiR-146a-3p underwent somatic mutations in 16.1% of analyzed specimens, mainly towards the deleterious C allele. In this report we propose a novel molecular marker of the clinical outcome of fvPTC patients. Rs2910164 increases the overall mortality with inhibition of NIS and disruption of radioiodine uptake as a possible mechanism.

SYNTHESIS AND BIOLOGICAL ACTIVITY OF NOVEL 6-PHENYL-1H-PYRROLO[3,4-c]PYRIDINE-1,3-DIONE DERIVATIVES.

The new pyrrolo[3,4-c]pyridines have been synthesized. 4-Methyl-6-phenyl-1H-pyrrolo[3,4-cpyridine-1,3-dione (1) was the key intermediate for the synthesis of the novel derivatives of various chemical structures. In the first step, the pyrrolo[3,4-c]pyridine-1,3-dione 1 was alkylated to the corresponding N-alkyl-4- methyl-6-phenyl-IH-pyrrolo[3,4-c]pyridine-1,3-dione derivatives 2a-f. The Mannich bases 3a-j were synthesized by treating pyrrolo[3,4-c]pyridine-1,3-dione 1 with appropriate amines and formaldehyde. Hydrolysis of ester 2a gave the appropriate acid 5. Next, amides 4a-e have been obtained. The structures of the new compounds were confirmed by elemental analysis, IR and NMR spectra. The antitumor and antimicrobial activities in vitro of the obtained derivatives were examined. Mannich bases 3c and 3g showed activity against C. albicans and S. aureus.

The diagnosis of nonclassic congenital adrenal hyperplasia due to 21-hydroxylase deficiency, based on serum basal or post-ACTH stimulation 17-hydroxyprogesterone, can lead to false-positive diagnosis.

OBJECTIVE: As nonclassic congenital adrenal hyperplasia (NCCAH) needs to be taken into account in women with hyperandrogenism, we aimed to assess whether the recommended level of poststimulated 17OHP ≥30 nmol/l confirms NCCAH. PATIENTS AND METHODS: Forty, consecutive women with biochemical and/or clinical hyperandrogenism (aged 25·4, 18-38) suspected of having NCCAH were recruited to the study. In patients with 17OHP level between 5·1 and 29·9 nmol/l an ACTH stimulation test was performed. In patients with basal or poststimulated 17OHP ≥30 nmol/l, twenty-four-hour urinary steroid profile (USP) analysis was performed and CYP21A2 mutation was assessed. In selected patients with poststimulated 17OHP <30 nmol/l USP was also performed. RESULTS: The group was divided into two subgroups with basal or poststimulated 17OHP ≥30 nmol/l (group A) and with poststimulated 17OHP <30 nmol/l (group B). Among 40 patients, basal or poststimulated 17OHP ≥30 nmol/l was found in 21, but NCCAH was confirmed by USP followed by genetic testing only in 5 (24%). Four patients were diagnosed as heterozygotes, and in twelve, no CYP21A2 mutation was detected. CONCLUSION: The diagnosis of NCCAH based only on serum 17OHP measurements (basal or poststimulated) may lead to false-positive diagnosis when performed by immunoassay with a cut-off value of ≥30 nmol/l. The definitive diagnosis can be established based on USP and/or genetic testing.

SYNTHESIS, ANTI-HEPATITIS B AND C VIRUS ACTIVITY AND ANTITUMOR SCREENING OF NOVEL THIAZOLO[4,5-D]-PYRIMIDINE DERIVATIVES.

The paper describes the synthesis, antivirus and antitumor evaluation of novel thiazolo[4,5-d]pyrim- idine derivatives. The target compounds 3a-h were synthesized by cyclocondensation of 4-amino-N'-(phenyl- methylidene)-3-phenyl-2-thioxo-2,3-dihydrothiazole-5-carbohydrazides 2a-d with aromatic aldehydes. The structures of new compounds were determined by IR, ¹H-NMR and elemental analysis. Thiazolopyrimidines 3a and 3d-h were screened by the National Institute of Allergy and Infectious Diseases against various viruses. Four compounds 3e-h showed in vitro anti-HCV activity. One (3e) demonstrated significant activity against HBV and was submitted to an anti-HBV in vivo assay but had a low bioavailability. As a result of antitumor study, compound 3h was found to be most potent against leukemia SR.

Variants in the ATM-CHEK2-BRCA1 axis determine genetic predisposition and clinical presentation of papillary thyroid carcinoma.

The risk of developing papillary thyroid carcinoma (PTC), the most frequent form of thyroid malignancy, is elevated up to 8.6-fold in first-degree relatives of PTC patients. The familial risk could be explained by high-penetrance mutations in yet unidentified genes, or polygenic action of low-penetrance alleles. Since the DNA-damaging exposure to ionizing radiation is a known risk factor for thyroid cancer, polymorphisms in DNA repair genes are likely to affect this risk. In a search for low-penetrance susceptibility alleles we employed Sequenom technology to genotype deleterious polymorphisms in ATM, CHEK2, and BRCA1 in 1,781 PTC patients and 2,081 healthy controls. As a result of the study, we identified CHEK2 rs17879961 (OR = 2.2, P = 2.37e-10) and BRCA1 rs16941 (odds ratio [OR] = 1.16, P = 0.005) as risk alleles for PTC. The ATM rs1801516 variant modifies the risk associated with the BRCA1 variant by 0.78 (P = 0.02). Both the ATM and BRCA1 variants modify the impact of male gender on clinical variables: T status (P = 0.007), N status (P = 0.05), and stage (P = 0.035). Our findings implicate an important role of variants in the ATM- CHEK2- BRCA1 axis in modification of the genetic predisposition to PTC and its clinical manifestations.

SYNTHESIS AND IN VITRO ANTIPROLIFERATIVE SCREENING OF NEW 2,7-NAPHTHYRIDINE-3-CARBOXYLIC ACID HYDRAZIDE DERIVATIVES.

The new pyrrolo[3,4-c]pyridines and 2,7-naphthyridine derivatives have been synthesized. 4-Hydroxy-8-methyl-l-oxo-6-phenyl-1,2-dihydro-2,7-naphthyridine-3-carboxylic acid hydrazide (4) was the key intermediate for the synthesis of the novel derivatives of various chemical structures: Schiff bases, 1,3,4-oxadiazoles, pyrazoles, carbohydrazides, semi- and thiosemicarbazides. The structures of these new compounds were confirmed by elemental analysis and IR, NMR and MS spectra. The antitumor activities of the obtained derivatives were examined. Eight of the twenty one newly synthesized compounds were qualified by the NCI (Bethesda, MD, USA) for in vito screening against 60 different human tumor cell lines. The most active proved to be the Schiff bases.

Mechanisms of action of thyroid hormones in the skeleton.

BACKGROUND: Thyroid hormones regulate skeletal development, acquisition of peak bone mass and adult bone maintenance. Abnormal thyroid status during childhood disrupts bone maturation and linear growth, while in adulthood it results in altered bone remodeling and an increased risk of fracture SCOPE OF REVIEW: This review considers the cellular effects and molecular mechanisms of thyroid hormone action in the skeleton. Human clinical and population data are discussed in relation to the skeletal phenotypes of a series of genetically modified mouse models of disrupted thyroid hormone signaling. MAJOR CONCLUSIONS: Euthyroid status is essential for normal bone development and maintenance. Major thyroid hormone actions in skeletal cells are mediated by thyroid hormone receptor α (TRα) and result in anabolic responses during growth and development but catabolic effects in adulthood. These homeostatic responses to thyroid hormone are locally regulated in individual skeletal cell types by the relative activities of the type 2 and 3 iodothyronine deiodinases, which control the supply of the active thyroid hormone 3,5,3'-L-triiodothyronine (T3) to its receptor. GENERAL SIGNIFICANCE: Population studies indicate that both thyroid hormone deficiency and excess are associated with an increased risk of fracture. Understanding the cellular and molecular basis of T3 action in skeletal cells will lead to the identification of new targets to regulate bone turnover and mineralization in the prevention and treatment of osteoporosis. This article is part of a Special Issue entitled Thyroid hormone signaling.

Alternative splicing of iodothyronine deiodinases in pituitary adenomas. Regulation by oncoprotein SF2/ASF.

Pituitary tumors belong to the group of most common neoplasms of the sellar region. Iodothyronine deiodinase types 1 (DIO1) and 2 (DIO2) are enzymes contributing to the levels of locally synthesized T3, a hormone regulating key physiological processes in the pituitary, including its development, cellular proliferation, and hormone secretion. Previous studies revealed that the expression of deiodinases in pituitary tumors is variable and, moreover, there is no correlation between mRNA and protein products of the particular gene, suggesting the potential role of posttranscriptional regulatory mechanisms. In this work we hypothesized that one of such mechanisms could be the alternative splicing. Therefore, we analyzed expression and sequences of DIO1 and DIO2 splicing variants in 30 pituitary adenomas and 9 non-tumorous pituitary samples. DIO2 mRNA was expressed as only two mRNA isoforms. In contrast, nine splice variants of DIO1 were identified. Among them, five were devoid of exon 3. In silico sequence analysis of DIO1 revealed multiple putative binding sites for splicing factor SF2/ASF, of which the top-ranked sites were located in exon 3. Silencing of SF2/ASF in pituitary tumor GH3 cells resulted in change of ratio between DIO1 isoforms with or without exon 3, favoring the expression of variants without exon 3. The expression of SF2/ASF mRNA in pituitary tumors was increased when compared with non-neoplastic control samples. In conclusion, we provide a new mechanism of posttranscriptional regulation of DIO1 and show deregulation of DIO1 expression in pituitary adenoma, possibly resulting from disturbed expression of SF2/ASF.

The assessment of sIgA, histatin-5, and lactoperoxidase levels in saliva of adolescents with dental caries.

BACKGROUND: Saliva contains a number of protective factors such as mucins, immunoglobulins (e.g., IgA, IgG, and IgM), and enzymes (e.g., lysozyme and lactoperoxidases) that play an important role in the maintenance of oral health. The aim of this study was to compare levels of sIgA, histatin-5, and lactoperoxidase in saliva of adolescents with dental caries. MATERIAL AND METHODS: Thirty-five adolescents (age 18 years) from high school were examined. Eight subjects with DMF=3 (Group I) and 27 adolescents with DMF>11 (Group II) were enrolled for this study. Clinical evaluation procedures comprised oral examination (including tooth, periodontal, and oral mucosal status) and collection of saliva samples. Saliva was collected for enzyme-linked immunosorbent assay (ELISA) and was used for determination of sIgA, histatin-5, and lactoperoxidase levels. RESULTS: Our results showed that adolescents with very high intensity of dental caries (DMF>11) had increased levels of sIgA, histatin-5, and lactoperoxidase compared to adolescents with lower intensity of caries. The increase was statistically significant (p<0.05). CONCLUSIONS: We suggest that high intensity of caries is associated with increased levels of some salivary components - sIgA, histatin-5 and lactoperoxidase - that possess strong bactericidal or bacteriostatic effects, resulting in aggregation of oral bacteria and their clearance from the oral cavity.

Mucin levels in saliva of adolescents with dental caries.

BACKGROUND: Human saliva, a complex secretion that contains a mixture of inorganic and organic molecules, plays an essential role in the maintenance of oral health. Mucins are the major macromolecular component of the secretion and are considered the first line of defense for epithelial tissues. The aim of this study was to compare levels of mucins (MUC5B, MUC7, and MUC1) in saliva of young subjects with dental caries. MATERIAL AND METHODS: All patients had DMF (decay/missing/filled) higher than value 0. Eight subjects with DMF=3 (control group) and 27 adolescents with DMF >11 (research group) were recruited for this study. Clinical evaluation procedures were oral examination, including tooth, periodontal, oral mucosal status, and collection of saliva samples. Saliva was collected for mucin assay. Enzyme-linked immunosorbent assay was used to quantitate MUC5B, MUC7, and MUC1. RESULTS: Our results indicate that adolescents with very high intensity of dental caries disease had increased levels of MUC1 and MUC5B. The membrane mucin MUC1 protein levels in the group with DMF>11 (research group) were higher compared to the group with DMF=3 (control group), and the increase was statistically significant (p=0.011). Similarly, secreted mucin MUC5B protein levels were higher (p=0.06) in the group with DMF>11 (research group). Although MUC7 protein levels were slightly reduced in symptomatic subjects, the decrease was statistically insignificant (p=0.918). CONCLUSIONS: Our data suggest links between the production of mucins, especially MUC1 and MUC5B in saliva, and dental caries disease.

Persistent hypercalcaemia associated with two pathogenic variants in the CYP24A1 gene and a parathyroid adenoma-a case report and review.

Introduction: 24-Hydroxylase, encoded by the CYP24A1 gene, is a crucial enzyme involved in the catabolism of vitamin D. Loss-of-function mutations in CYP24A1 result in PTH-independent hypercalcaemia with high levels of 1,25(OH)2D3. The variety of clinical manifestations depends on age, and underlying genetic predisposition mutations can lead to fatal infantile hypercalcaemia among neonates, whereas adult symptoms are usually mild. Aim of the study: We report a rare case of an adult with primary hyperparathyroidism and loss-of-function mutations in the CYP24A1 gene and a review of similar cases. Case presentation: We report the case of a 58-year-old woman diagnosed initially with primary hyperparathyroidism. Preoperatively, the suspected mass adjoining the upper pole of the left lobe of the thyroid gland was found via ultrasonography and confirmed by 99mTc scintigraphy and biopsy as the parathyroid gland. The patient underwent parathyroidectomy (a histopathology report revealed parathyroid adenoma), which led to normocalcaemia. After 10 months, vitamin D supplementation was introduced due to deficiency, and the calcium level remained within the reference range. Two years later, biochemical tests showed recurrence of hypercalcaemia with suppressed parathyroid hormone levels and elevated 1,25(OH)2D3 concentrations. Further investigation excluded the most common causes of PTH-independent hypercalcaemia, such as granulomatous disease, malignancy, and vitamin D intoxication. Subsequently, vitamin D metabolites were measured using LC-MS/MS, which revealed high levels of 25(OH)D3, low levels of 24,25(OH)2D3 and elevated 25(OH)2D3/24,25(OH)2D3 ratios, suggesting a defect in vitamin D catabolism. Molecular analysis of the CYP24A1 gene using the NGS technique revealed two pathogenic variants: p.(Arg396Trp) and p.(Glu143del) (rs114368325 and rs777676129, respectively). Conclusions: The diagnostic process for hypercalcaemia becomes complicated when multiple causes of hypercalcaemia coexist. The measurement of vitamin D metabolites using LC-MS/MS may help to identify carriers of CYP24A1 mutations. Subsequent molecular testing may contribute to establishing the exact frequency of pathogenic variants of the CYP24A1 gene and introducing personalized treatment.

Free-Circulating Nucleic Acids as Biomarkers in Patients After Solid Organ Transplantation.

A number types of extracellular DNA (eg, cell-free, cfDNA) circulate in human blood, including mitochondrial, transcriptome, and regulatory DNA, usually at low concentrations. Larger amounts of cfDNA appear in any inflammatory condition, including organ damage due to a variety of reasons. The role of cfDNA in solid organ transplantation is discussed in this review as a valuable additional tool in the standard of care of transplant patients. Post-transplant monitoring requires the use of high-quality biomarkers for early detection of graft damage or rejection to be able to apply early therapeutic intervention. CfDNA complements the traditional monitoring strategies, being a risk stratification tool and an important prognostic marker. However, improving the sensitivity and specificity of cfDNA detection is necessary to facilitate personalized patient management, warranting further research in terms of measurement, test standardization, and storage, processing, and shipping. A diagnostic test (Allosure, CareDx, Inc., Brisbane, CA) for kidney, heart and lung transplant patients is now commercially available, and validation for other organs (eg, liver) is pending. To date, donor-derived cfDNA in combination with other biomarkers appears to be a promising tool in graft rejection as it is minimally invasive, time-sensitive, and cost-effective. However, improvement of sensitivity and specificity is required to facilitate personalized patient management. Whether it could be an alternate to graft biopsy remains unclear.

Circular RNAs Could Encode Unique Proteins and Affect Cancer Pathways.

circRNAs constitute a novel class of RNA, generally considered as non-coding RNAs; nonetheless, their coding potential has been under scrutiny. In this work, we systematically explored the predicted proteins of more than 160,000 circRNAs detected by exome capture RNA-sequencing and collected in the MiOncoCirc pan-cancer compendium, including normal and cancer samples from different types of tissues. For the functional evaluation, we compared their primary structure and domain composition with those derived from the same linear mRNAs. Among the 4362 circRNAs potentially encoding proteins with a unique primary structure and 1179 encoding proteins with a novel domain composition, 183 were differentially expressed in cancer. In particular, eight were associated with prognosis in acute myeloid leukemia. The functional classification of the dysregulated circRNA-encoded polypeptides showed an enrichment in the heme and cancer signaling, DNA-binding, and phosphorylation processes, and disclosed the roles of some circRNA-based effectors in cancer.

[Dental caries of the developmental age as a civilization disease]. / Próchnica wieku rozwojowego choroba cywlizacyjna.

According to the definition of the World Health Organization (WHO), dental caries is a local pathological process of the extrasomatic background, leading to enamel decalcification, decomposition of dental hard tissue, and in consequence to formation of a dental cavity. Morbidity of dental caries increases with age, reaching 100% of children, aged from 6 to 7. Poland is one of few European countries where the incidence of dental caries in children did not decrease, despite recommendations of WHO for 2000 year, aimed at the decrease in the incidence of dental caries among 6-year-old children to the level of 50%. The recommendation of WHO for 2015 year is to reduce the incidence of dental caries to 30% among 6-year-olds, i.e., 70% of 6 year-old children should be free of dental caries. Apart from genetic conditioning, inappropriate health behaviors, nutritional habits and gastroesophageal reflux disease influence the development of dental caries. Consumption of 'fast food' and drinking sweetened beverages of low pH contribute markedly to the development of dental caries, decreasing simultaneously consumption of pro healthy foods, including milk and cereals. Taking into consideration perspective clinical examinations of children and adolescents, evaluating the relationship between dental caries and nutritional habits as well as environmental conditioning, the study shows current data about factors, contributing to the incidence of dental caries in children, collected from the literature. The attention was paid to the relationship between dental caries and gastroesophageal reflux disease and the necessity of its early diagnostics and proper treatment.

[Selected eating habits and caries occurrence in adolescents]. / Wybrane zachowania zywieniowe a wystepowanie próchnicy U Mlodziezy.

INTRODUCTION: Normal nutrition is an important element of caries prophylaxis. The aim of the study was to evaluate the influence of chosen bad eating habits on caries occurrence in adolescents. MATERIAL AND METHODS: The examination was carried out in the group of 367 (68.5% girls and 31.5% boys) pupils of the 3rd year of high schools in Bialystok in 2011. Own questionnaire was used for the evaluation of eating habits. Caries intensity (PUW) was assessed by dental examination. To verify statistic hypotheses, the level of significance was estimated at p < 0.05. All calculation was performed with the use of the Statistic programme Statistica 7.0 (StatSoft). RESULTS: It was stated that 5 meals a day were consumed by 29.4% of examined pupils, 4 meals were eaten by 33.2%, 3 meals--by 31.3%, 2 meals--by 5.4%, and one meal was consumed by 0.5% of pupils. There were 55.3% of pupils who had irregular meals while 24.3% did not eat breakfast and 62.9%--did not have elevenses. Eating between meals was stated in 96.7% of pupils. Sweets consumption was revealed as follows: 16.8% several times a day, 19%--once a day, 40.5%--several times a week, 18.5%--several times a month, 5.2%--rarely or never. "Fast food" was not consumed by 34% of examined pupils while 54.6% consumed such meals several times a month, 10.9%--several times a week, and 0.5%--once a day. Higher level of caries intensity was observed in girls (mean coefficient PUW = 11.3) than in boys (mean coefficient PUW = 10.8). Mean coefficient PUW = 11.5 concerned pupils with consumption of fast food several times a month while PUW = 11.0 in those who consumed it several times a week, and PUW = 10.7 in pupils who eat rarely or never such food. As far as eating sweets, PUW was higher than mean (12.4) in pupils who eat sweets several times a day. Persons who eat between meals were characterized by higher coefficient (PUW = 13.0) as compared to those who eat between meals rarely (PUW = 11.3) and those who do not eat between meals (PUW = 8.9). CONCLUSIONS: (1) Bad eating habits (irregular meals, skipping breakfast, eating between meals as well as overeating sweets) in the developmental age can be a significant caries coefficient in adolescents. 2. High mean coefficient PUW in the examined group of 18-year-old pupils, in relation to abnormal eating habits in the significant percentage of those pupils, indicates the necessity of health education as far as caries prophylaxis is concerned.

Structure and cytotoxic properties of 1-hydroxy-5-methyl-7-phenylpyrido[3,4-d]pyridazin-4(3H)-one and its mono- and disubstituted ethyl acetates.

Derivatives of pyrido[3,4-d]pyridazine, namely, 1-hydroxy-5-methyl-7-phenylpyrido[3,4-d]pyridazin-4(3H)-one dimethylformamide monosolvate, C14H11N3O2·C3H7NO (2), ethyl [1-(2-ethoxy-2-oxoethoxy)-5-methyl-4-oxo-7-phenyl-3,4-dihydropyrido[3,4-d]pyridazin-3-yl]acetate, C18H17N3O4 (3), and ethyl [(5-methyl-4-oxo-7-phenyl-3,4-dihydropyrido[3,4-d]pyridazin-1-yl)oxy]acetate, C22H23N3O6 (4), were synthesized with the aim of discovering new potential biologically active agents. The properties of all three derivatives were characterized by 1H NMR, 13C NMR and FT-IR spectroscopic analysis. All the crystals were obtained by a solvent diffusion method from dimethylformamide (DMF) or dimethyl sulfoxide (DMSO) and characterized by single-crystal X-ray diffraction. The collected X-ray data revealed that the crystals of 2 and 4 belong to the triclinic space group P-1, whereas the crystal of 3 belongs to the monoclinic space group P21/c. The presented derivatives crystallized with one molecule in the asymmetric unit, but only compound 2 crystallized as a solvate with DMF. Structure analysis showed that the molecule of 2 exists as its amide-imidic acid tautomer and that O-alkylation occurred before N-alkylation during the synthesis of the mono- and disubstituted derivatives, i.e. 3 and 4, respectively. The molecular geometries of the 5-methyl-7-phenylpyrido[3,4-d]pyridazine core within the studied derivatives differ in the mutual orientation of the rings. The interplanar angles between the heterocyclic ring and the bound aromatic ring are 1.71 (7), 18.16 (3) and 3.1 (1)° for 2, 3 and 4, respectively. The potential cytotoxicity of these compounds was evaluated against one normal (HaCat) and four human cancer cell lines (A549, DU145, MDA-MB-231 and SKOV-3).

The Object Segmentation from the Microstructure of a FSW Dissimilar Weld.

Friction stir welding (FSW) is an environmentally friendly, solid-state welding technique. In this research work, we analyze the microstructure of a new type of FSW weld applying a two- stage framework based on image processing algorithms containing a segmentation step and microstructure analysis of objects occurring in different layers. A dual-speed tool as used to prepare the tested weld. In this paper, we present the segmentation method for recognizing areas containing particles forming bands in the microstructure of a dissimilar weld of aluminum alloys made by FSW technology. A digital analysis was performed on the images obtained using an Olympus GX51 light microscope. The image analysis process consisted of basic segmentation methods in conjunction with domain knowledge and object detection located in different layers of a weld using morphological operations and point transformations. These methods proved to be effective in the analysis of the microstructure images corrupted by noise. The segmentation parts as well as single objects were separated enough to analyze the distribution on different layers of the specimen and the variability of shape and size of the underlying microstructures, which was not possible without computer vision support.