Summary Mammalian tissues calcify with age and injury. Analogous to bone formation, osteogenic cells are thought to be recruited to the affected tissue and induce mineralization. In the heart, calcification of cardiac muscle leads to conduction system disturbances and is one of the most common pathologies underlying heart blocks. However the cell identity and mechanisms contributing to pathological heart muscle calcification remain unknown. Using lineage tracing, murine models of heart calcification and in vivo transplantation assays, we show that cardiac fibroblasts (CFs) adopt an osteoblast celllike fate and contribute directly to heart muscle calcification. Small molecule inhibition of ENPP1, an enzyme that is induced upon injury and regulates bone mineralization, significantly attenuated cardiac calcification. Inhibitors of bone mineralization completely prevented ectopic cardiac calcification and improved post injury heart function. Taken together, these findings highlight the plasticity of fibroblasts in contributing to ectopic calcification and identify pharmacological targets for therapeutic development.

We analyzed proliferative index of myeloma plasmocytes (PC-PI) in a cohort of 217 patients with multiple myeloma (MM) treated with conventional chemotherapy and biological agents, thalidomide and bortezomib. In the whole group was a difference between overall survival (OS) favoring patients with PC-PI < 2.8% (median overall survival 30 vs 12 months) with a borderline significance (p = 0.06). However, after approximately 40 months from diagnosis the curves merged, suggesting the influence of novel drugs. In patients treated with conventional chemotherapy only, the difference maintained significant even after 40 months (median overall survival 25 vs 10months, p = 0.015), whereas in the group treated with thalidomide and bortezomib was no difference, with medians over 39 months. Even patients with low PC-PI profited from the treatment with novel drugs. Presented results suggest that the treatment of MM with novel agents overcomes the prognostic significance of PC-PI and should be used in all MM patients.

In a group of 117 patients with multiple myeloma (MM) examined at the time of diagnosis, i.e. excluding previous chemotherapy, we analysed the levels of propidium-iodide (proliferative) -PC-PI/CD 138 and annexin-V (apoptotic) -PC-AI/CD 138 indices of myeloma plasmocytes using the method of flow-cytometry to determine their relationship to prognosis. It was revealed that patients with high values of PC-PI/CD 138 had substantially worse overall survival than those with low values. Patients with a level of propidium-iodide index ≥ 2,8 % exprimed a median survival of 13 months only in comparison with 42 months in patients with levels < 2,8 % (p = 0,0005). In the PC-AI/CD 138 index a reverse trend was registered. Patients with PC-AI/CD 138 ≥ 4,0 % had long overall survival (median was not assessable at the time of evaluation), whereas patients with low apoptosis values < 4,0 % had median overall survival 16 months only (p = 0,01). Based on the sequentional graphic analysis of the curves of overall survival was found that the optimal discrimination level sequestering patients with good and poor prognosis was, in the case of PC-PI/CD 138 value 2,8 %, whereas in the case of PC-AI/CD 138 value 4,0 %. Among patients with good prognosis, there were no statistically significant differences in overall survival according to different levels of proliferative and apoptotic index. We conclude that evaluation of the propidium-iodide and annexin-V index using flow-cytometry is a quick, useful, and easily accessible method for the evaluation of plasma cell kinetics and thus prognosis of the disease, multiple myeloma.

Aim Extremely low birthweight (ELBW) neonates require a high protein intake, but this can be challenging in the very rare cases when they also have phenylketonuria (PKU). This is due to a lack of suitable parenteral nutrition or enteral formula. Our aim was to analyse tolerance to phenylalanine in these infants. Material There are approximately 110 000 children born in the Czech Republic each year. A neonatal screening programme from 2005 to 2020 found that 320 neonates had PKU, including 30 premature neonates with a birth weight of less than 2500 g. Results This study focused on three neonates who were born with ELBWs of 720, 740 and 950 g, respectively. Phenylalanine levels normalised in ELBW neonates with PKU within 1 week of the introduction of low‐phenylalanine parenteral or enteral nutrition. The tolerance to phenylalanine was very high (70–110 mg/kg) in the first months of life, due to a rapid weight gain, but significantly decreased during infancy. Conclusion Extremely low birthweight neonates with PKU need special dietary management. Regular assessments of phenylalanine are necessary during the first weeks of life to allow prompt dietary adjustments that reflect rapid weight gain and transitory high tolerance to phenylalanine.

Background: Autologous stem cell transplantation (ASCT) plays an important role in the treatment of multiple myeloma (MM) patients (pts). In this report, we describe the longterm outcome of cohort of 185 pts with newly diagnosed symptomatic MM treated with ASCT. Median follow-up from the start of therapy was 102.8 months (range 67.2–150.4). We have specifically analyzed benefit from the newer drugs used in the relapsed setting. Methods: A total of 185 MM pts underwent ASCT in the clinical trial 4W of Czech Myeloma Group in 18 centres in Czech Republic and Slovak Republic between 1996 and 2001. The conditioning regimen was high dose melphalan (200mg/m2) in all pts. At diagnosis of MM 72.4% (134/185) of pts had stage III according to Durie- Salmon, 18.4% (34/185) stage II and 9.2% (17/185) stage I. Clinical stages according to ISS were the following: stage 1 in 42.5% (74/174) of pts, stage 2 in 36.8% (64/174) and stage 3 in 20.7% (36/174) pts. Types of monoclonal immunoglobulin were as following: 67.6% (125/185) IgG, 21.1% (39/185) IgA, 9.7% (18/185) light chain, 1.6% (3/185) IgD. Renal insufficiency was presented in 5.9% (11/185) of pts. Median age at transplantation was 54.8 years (range: 28.3–69.2). When the symptomatic relapse of MM after ASCT was occurred, 34.6% (45/130) of pts were treated by conventional chemotherapy (CC) alone, 22.3% (29/130) by thalidomide based regimen, 10.7% (14/130) by bortezomib based regimen, 22.3% (29/130) pts underwent re-transplantation and 10% (13/130) of pts received combination of newer drugs and re-transplantation. Results: Following ASCT, overall response rate (ORR) was 93.5% (173/185), 29.2% (54/185) of pts were in CR, 38.4% (71/185) of pts were in VGPR, 25.9% (48/185) of pts in PR. Median of overall survival (OS) from start of treatment was 77.9 months (9.6 – 147.3), median of TTP was 39.8 months (7.0–146.9). Total of 23.2% (43/185) of pts are alive and disease free, 20.5% (38/185) of pts are alive with relapse and 56.3% (104/185) of pts died with median follow-up 8.3 years from the start of therapy. Significant prognostic parameters for better OS after ASCT were: ISS stage < III (p<0.001), achievement of CR after ASCT (p<0.001) and the use of the newer drugs in the relapsed setting. Patients treated with thalidomide and/or bortezomib comparing to pts treated with CC only had significantly longer survival (p<0.001). Patients treated with re-transplantation only had not better OS (46.1 vs 37.8 months; p=0.224) comparing to pts treated with CC. In opposite, pts treated with the newer drugs and re-transplantation had better prognosis (OS 85.7 vs 37.8 months; p<0.001) than pts treated by CC. In multivariate analysis thalidomide and/or bortezomib treatment in the relapse was the strongest factor for long-term survival (beta 1.988; beta (exp) 7.303; CI: 2.860–18.645; p<0.001) Conclusion: ASCT is effective procedure in MM pts. The achievements CR after transplantation, no presence of stage 3 according to ISS were significant parameters for long-term postransplant survival. Accessibility of the thalidomide and bortezomib was independent of other prognostic factors. The use of the newer drugs in the relapsed setting significantly improved prognosis of patients.