The Mediterranean diet (MD) is considered one of the healthiest dietary patterns. The aim of this study was to assess MD adherence in children and youth living in the Mediterranean region in Croatia and evaluate the differences in adherence to the MD among different educational stages. In total, 2722 individuals aged 2 to 24 years were enrolled in this study. Subjects were divided into different groups according to the Croatian educational system. Mediterranean Diet Quality Index (KIDMED) was used to assess adherence to the MD. In the total sample, the adherence to the MD was poor in 19.2%, average in 60.8%, and good in 20.1% of the study participants. The prevalence rate of poor adherence to the MD increased with higher educational stage, i.e., the highest prevalence rate of poor MD adherence was observed for college students (39.3%). Children having a higher number of snacks on days-off, those with lower physical activity, and not having breakfast together with a family are more likely to have poor MD adherence, while children having a higher number of snacks on working days are less likely to have a poor MD. The results of this study showed low adherence to the principles of the MD, confirming the need for improvement of adherence to the MD pattern in the studied population.

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Objectives: To assess genotoxic and cytotoxic effect of commercially available toothpastes with the different whitening ingredients. Materials and Methods: In vivo assessment of cytotoxicity and genotoxicity of whitening toothpastes with different ingredients using a buccal micronucleus cytome assay (BMCyt assay) comprised 199 participants randomly divided into ten groups based on used whitening or control/conventional toothpaste. The exfoliated buccal mucosal cells were collected, stained and microscopically evaluated at baseline, 30 days and 60 days after the beginning of treatment, and 30 days after completing treatment: follow-up. Results: The genotoxic parameters showed no biologically significant changes in any of the observed period for the tested toothpastes, while cytotoxic parameters (number of cells with karyorrhexis and condensed chromatin) showed statistically significant difference (p>0.05) amongst evaluation periods for the three peroxide containing toothpaste. Conclusions: Peroxide containing whitening toothpastes present significant increase of cytotoxicity (numbers of karyorrhexis and condensed chromatin) during the application period only. Clinical Significance: Whitening toothpaste show no genotoxic effect, while peroxide containing whitening toothpastes may present significant increase of cytotoxicity (numbers of karyorrhexis and condensed chromatin) during application period. In clinical conditions, the obtained changes can’t be considered as significant. Trial registration: ClinicalTrials.gov: NCT04460755

We describe two patients with diagnosis of chronic lymphocytic leukemia (CLL) in whom interphase fluo­rescence in situ hybridization (FISH) analysis revealed trisomy 12 and del(13)(q14.3) occurring in the same clone. These abnormalities are rarely seen together and the prognostic relevance of their coexistence is still unclear. According to some data, a probable adverse prognosis for this group of patients is suggested. Our patients have been in a stable phase of the disease for more than one year since the given abnormalities were documented in their karyotypes. Further study is necessary to determine the prognostic significance of coexistence of these abnormalities in CLL patients

CD44 expressed in monocytes and lymphocytes seems to play a crucial role in gastrointestinal inflammation, such as the one occurring in the context of inflammatory bowel diseases. Differentially methylated genes are distinctly expressed across monocyte subpopulations related to the state of Crohn’s disease. Hence, the aim of this study was to detect CD44 expression in leukocyte subpopulations in relation to the type of IBD, therapy, and disease duration. Monocyte subpopulations CD14++CD16−, CD14++CD16++, and CD14+CD16+ as well as other leukocytes were analyzed for their CD44 expression using flow cytometry in 46 patients with IBD and 48 healthy controls. Patients with Crohn’s disease treated with non-biological therapy (NBT) exhibited a lower percentage of anti-inflammatory CD14+CD16++ monocytes, whereas NBT-treated patients with ulcerative colitis had lower expression of CD44 on CD14+CD44+ lymphocytes in comparison to controls, respectively. Conversely, patients with Crohn’s disease treated with biological therapy had a higher percentage of CD44+ granulocytes but lower expression of CD44 on anti-inflammatory monocytes compared to controls. Median fluorescence intensity (MFI) of CD44 on CD44+CD14+ lymphocytes was higher in ulcerative colitis patients treated with biological therapy compared to NBT. The percentage of classical CD14++CD16− monocytes was lower in the <9 years of IBD duration subgroup compared with the longer disease duration subgroup. The present study addresses the putative role of differentiation and regulation of leukocytes in tailoring IBD therapeutic regimes.

The Mediterranean diet (MD) is known to be one of the healthiest dietary patterns. Despite the significance of a healthful diet during the early stage of life, data for young individuals indicate that nutrition problems are common. This cross-sectional study aimed to determine parental factors associated with MD adherence in children and adolescents living in the Mediterranean region in Croatia. In total, 2623 children aged 2 to 18 years and their parents participated in this study. Data were collected during the period from September 2021 to February 2022 by using an anonymous questionnaire. We used KIDMED and MEDAS questionnaires for assessing MD adherence in young individuals and their parents, respectively. To assess the association of children’s MD adherence categories with the parental predictors, we performed multivariate multinomial logistic regression. Results showed that the children of parents with a low MD adherence are much more likely to have poor MD adherence than good (OR = 47.54 (95% C.I 18.24, 123.87), p < 0.001) or average (OR = 5.64 (95% C.I 3.70, 8.6), p < 0.001) MD adherence. Further, children of fathers with higher BMI (OR = 1.035 (95% C.I 1.0, 1.071)) and those who do not live with both parents (OR = 1.703 (95% C.I 0.994, 2.916), p = 0.053) are more likely to have poor MD adherence than good MD adherence. These results indicate that interventions focusing on enhancing the quality of both parents’ diets could effectively improve their children’s eating habits.

CD44 expressed in monocytes and lymphocytes seems to play a crucial role in gastrointestinal inflammation, such as the one occurring in the context of inflammatory bowel diseases (IBD). Differentially methylated genes are distinctly expressed across monocyte subpopulations related to the state of Crohn's disease. Hence, the aim of this study was to detect CD44 expression at monocyte subpopulations, lymphocytes and granulocytes in relation to the type of IBD, therapy and disease duration. Monocyte subpopulations CD14++CD16−, CD14+CD16++ and CD14+CD16+, as well as other leukocytes, were analyzed for their CD44 expression using flow cytometry in 46 patients with IBD and 48 healthy controls. Patients with Crohn's disease treated with non-biological therapy (NBT) exhibited lower percentage of anti-inflammatory CD14+CD16++ monocytes, whereas NBT-treated patients with ulcerative colitis had lower expression of CD44 on CD14+CD44+ lymphocytes, in comparison to controls, respectively. Conversely, patients with Crohn's disease treated with biological therapy had higher percentage of CD44+ granulocytes, but lower expression of CD44 on anti-inflammatory monocytes compared to controls. Percentage of classical CD14++CD16- monocytes was lower in the <9 years of IBD duration subgroup, compared with the longer disease duration subgroup. The present study addresses the putative role of differentiation and regulation of monocyte and lymphocyte cells in tailoring IBD therapeutic regimes.

Continuous human malignant hematopoietic cell lines are invaluable tool for hematological research. Here we report on a new erythroleukemic cell line termed VES that was established from the bone marrow mononuclear cells (BMNC) of a 22-year-old woman with Ph+ CML during her second post-transplant period. Actually, after unsuccessful allo-transplantation, the patient received auto-transplant that also failed to engraft. At the time of initiation of BMNC into culture for the analysis of stromal cell capacity, her bone marrow was tested Ph-negative by both FISH and PCR. Instead of establishment of an adherent stromal layer in vitro, blast-like cells appeared in the supernatant on day 27. These blast cells expressed highly amplified bcr/abl locus as shown by FISH, whereas the patient’s bone marrow was still Ph-negative at the time. Following a very short period, the cultured cells started to express stable features of erythroleukemia cell line. Optimal growth was obtained by suspending the cells at concentration of 0.3x106 cells/ml in IMDM containing 10% FBS, 1% penicillin-streptomycin and 1% L-glutamine solution. After 3 days of culture, cell concentration varied between 1.3 and 1.8x106 cells/ml. VES cells were tested negative for the presence of EBV virus. For analysis of clonogenicity, VES cells at concentration of 103/ml were grown in a serum-free methylcellulose medium without cytokines (H4236; StemCell Tech., Canada). Following 7-day culture, 103 VES cells produced 251 ±42 and after 14 days 431 ±30 GF-independent colonies. Furthermore, on day 14 of the cell culture it was possible to observe reddish color of the colonies indicating the presence of hemoglobin. Cytological examination of cytospin preparations indicated homogenous population of leukemia blasts (>80%) the majority of them (>90%) being glycophorinA positive and MPO negative. Imunophenotyping and multiparameter flow cytometry revealed proerythroblastic lineage-associated profile: GlyA/CD235a+, CD11b+, CD15+, CD29+, CD33+, and CD117+. Conventional cytogenetics revealed complex karyotype with multiple numerical/structural abnormalities (MAKA), while metaphase FISH revealed the following aberrations: t(9;22), 5q31, 7q31, +8, +6. Since FISH analysis detected highly amplified bcr/abl locus, we tested VES cells’ sensitivity to imatinib (Gleevec). Treatment with imatinib for 3 days (MTS assay) inhibited proliferation of VES cells with IC50 value of 0.2mM. Following 14 days of culture in methylcellulose medium with addition of 0.2mM imatinib, the clonogenic potential of VES cells was reduced by 55% in relation to untreated control. We demonstrated that imatinib induced apoptosis in VES cells in time- and dose-dependant manner, assessed with AnnexinV and PI staining, while there were no significant changes observed in the cell cycle except for a mild increase in cells in G0/G1 phase. Although further analyses are required, we believe that VES cells represent a new Ph+ erythroleukemia cell line and a suitable model for studying Ph+ malignant hematopoiesis in vitro.