The correlation of mutations and expressions of genes within the PI3K/Akt/mTOR pathway in breast cancer : a preliminary study

There is an urgent need to seek new molecular biomarkers helpful in diagnosing and treating breast cancer. In this elaboration, we performed a molecular analysis of mutations and expression of genes within the PI3K/Akt/mTOR pathway in patients with ductal breast cancer of various malignancy levels. We recognized significant correlations between the expression levels of the studied genes. We also performed a bioinformatics analysis of the data available on the international database TCGA and compared them with our own research. Studies on mutations and expression of genes were conducted using High-Resolution Melt PCR (HRM-PCR), Allele-Specific-quantitative PCR (ASP-qPCR), Real-Time PCR molecular methods in a group of women with ductal breast cancer. Bioinformatics analysis was carried out using web source Ualcan and bc-GenExMiner. In the studied group of women, it was observed that the prevalence of mutations in the studied PIK3CA and AKT1 genes was 29.63%. It was stated that the average expression level of the PIK3CA, PIK3R1, PTEN genes in the group of breast cancer patients is lower in comparison to the control group, while the average expression level of the AKT1 and mTOR genes in the studied group was higher in comparison to the control group. It was also indicated that in the group of patients with mutations in the area of the PIK3CA and AKT1 genes, the PIK3CA gene expression level is statistically significantly lower than in the group without mutations. According to our knowledge, we demonstrate, for the first time, that there is a very strong positive correlation between the levels of AKT1 and mTOR gene expression in the case of patients with mutations and without mutations

Ultrastructural visualization of 3D chromatin folding using volume electron microscopy and DNA in situ hybridization.

The human genome is extensively folded into 3-dimensional organization. However, the detailed 3D chromatin folding structures have not been fully visualized due to the lack of robust and ultra-resolution imaging capability. Here, we report the development of an electron microscopy method that combines serial block-face scanning electron microscopy with in situ hybridization (3D-EMISH) to visualize 3D chromatin folding at targeted genomic regions with ultra-resolution (5 × 5 × 30 nm in xyz dimensions) that is superior to the current super-resolution by fluorescence light microscopy. We apply 3D-EMISH to human lymphoblastoid cells at a 1.7 Mb segment of the genome and visualize a large number of distinctive 3D chromatin folding structures in ultra-resolution. We further quantitatively characterize the reconstituted chromatin folding structures by identifying sub-domains, and uncover a high level heterogeneity of chromatin folding ultrastructures in individual nuclei, suggestive of extensive dynamic fluidity in 3D chromatin states

Validation of selected medical centers involved in molecular diagnostics of cancer in the field of EGFR1 mutations determination

Rak płuca to jeden najczęściej występujących nowotworów złośliwych w Polsce i na świecie. Według danych statystycznych rocznie jest przyczyną zgonów 1,3 miliona osób na całym świecie. W niedrobnokomórkowym raku płuc (NDRP) u niektórych chorych (10–15% Ameryka Północna i Europa Zachodnia; 30–35% Japonia i Wschodnia część Azji) występują mutacje somatyczne w genie receptora nabłonkowego czynnika wzrostu (EGFR) powodujące stałą aktywność tego receptora. Występowanie takich mutacji ściśle wiąże się ze skutecznością działania niektórych inhibitorów kinazy tyrozynowej (TKI). Określenie statusu genu EGFR jest kluczowe w doborze najbardziej odpowiedniego schematu leczenia chorych z NDRP. Celem tej pracy jest wykazanie, czy ośrodki zaangażowane w diagnostykę molekularną chorób nowotworowych posiadają odpowiedni potencjał, aby skutecznie przeprowadzić analizę statusu genu EGFR1 w zakresie zmian w eksonach 19. i 21. Dodatkowym celem jest wypracowanie odpowiednich standardów postępowania. Wynikiem przeprowadzonego procesu walidacji są następujące zalecenia dla laboratoriów diagnostycznych: 1. Materiał do izolacji DNA powinien zawierać nie mniej niż 50% utkania nowotworowego; 2. Ujednolicenie procedury izolacji DNA ze skrawków parafinowych wymaga stosowania gotowego zestawu do izolacji DNA; 3. W przypadku braku jednoznacznego wyniku, powinno się wykorzystać dwie metody oznaczania mutacji, zaleca się, aby jednego z wykonywanych oznaczeń dokonać przy wykorzystaniu metody sekwencjonowania bezpośredniego; 4. Zaleca się rozszerzenie panelu analizowanych eksonów do 18., 19., 20. i 21.; 5. Od momentu wypisania skierowania na badanie diagnostyczne do momentu przekazania wyniku badania nie powinno upłynąć więcej niż 10 dni roboczych. Onkol. Prak. Klin. 2011; 7, 3: 138–145Lung cancer is one of the most common cancers in Poland and abroad. According to statistics, it causes the death of 1.3 million people per year worldwide. In a nonsmall cell lung cancer (NSCLC), some patients have somatic mutations in the gene for epidermal growth factor receptor (EGFR), resulting in a constant activity of this receptor (10–15% patients of North American and Western European origin, and 30–35% of patients from Japan and Eastern Asia). The occurrence of such mutations is closely associated with efficacy of tyrosine kinase inhibitors (TKI). Thus, determination of EGFR status is crucial in selecting the most appropriate treatment of patients with NSCLC. The aim of this paper is to show whether the laboratories involved in molecular diagnostics for cancer have the potential to effectively determinate the mutation status (in exons 19 and 21) of the EGFR1 gene. An additional objective is to develop appropriate standards for mutation testing in non-small cell lung cancer. As the result of valiadtion process conducted in the study, the following recommendations for diagnostic laboratories were approved: at least 50% of cancer cells should be present in a tissue for DNA isolation; 2. The method of DNA isolation should be standardized, the most appropriate is usage of DNA isolation kits; 3. In case of equivocal results two independent molecular methods should be employed, one of them should be direct sequencing; 4. It is recommended to extend the panel of analyzed exons to 18, 19, 20 and 21; 5 The turnaround time (TAT) should not take more than 10 working days Onkol. Prak. Klin. 2011; 7, 3: 138–14

Expert opinion of the Heart Failure Association of the Polish Society of Cardiology, the College of Family Physicians in Poland, and the Polish Society of Family Medicine on the peri discharge management of patients with heart failure

Despite advances in the treatment of heart failure (HF), the rate of hospitalisation for exacerbations of the disease remains high. One of the underlying reasons is that recommended guidelines for the management of HF are still too rarely followed in daily practice. Disease exacerbation requiring inpatient treatment is always a factor that worsens the prognosis, and thus signals disease progression. This is also a key moment when therapy should be modified for HF exacerbation, or initiated in the case of newly diagnosed disease. Inpatient treatment and the peri‑discharge period is the time when the aetiology and mechanism of HF decompensation should be established. Therapy should be individualised based on aetiology, HF phenotype, and comorbidities; it should take into account the possibilities of modern treatment. According to the recommendations of the European Society of Cardiology (ESC), patients with HF should receive multidisciplinary management. Cooperation between the various members of the multidisciplinary team taking care of patients with HF improves the efficiency and quality of treatment. This document expands and details the information on the peri‑discharge management of HF contained in the 2021 ESC guidelines and the 2022 American Heart Association (AHA)/American College of Cardiology (ACC)/Heart Failure Society of America (HFSA) guidelines

Testing CPT symmetry in ortho-positronium decays with positronium annihilation tomography

Charged lepton system symmetry under combined charge, parity, and time-reversal transformation (CPT) remains scarcely tested. Despite stringent quantum-electrodynamic limits, discrepancies in predictions for the electron–positron bound state (positronium atom) motivate further investigation, including fundamental symmetry tests. While CPT noninvariance effects could be manifested in non-vanishing angular correlations between final-state photons and spin of annihilating positronium, measurements were previously limited by knowledge of the latter. Here, we demonstrate tomographic reconstruction techniques applied to three-photon annihilations of ortho-positronium atoms to estimate their spin polarisation without magnetic field or polarised positronium source. We use a plastic-scintillator-based positron-emission-tomography scanner to record ortho-positronium (o-Ps) annihilations with single-event estimation of o-Ps spin and determine the complete spectrum of an angular correlation operator sensitive to CPT-violating effects. We find no violation at the precision level of 10−4, with an over threefold improvement on the previous measurement

From tests of discrete symmetries to medical imaging with J-PET detector

We present results on CPT symmetry tests in decays of positronium performed with the precision at the level of 10$^{-4}$, and positronium images determined with the prototype of the J-PET tomograph. The first full-scale prototype apparatus consists of 192 plastic scintillator strips readout from both ends with vacuum tube photomultipliers. Signals produced by photomultipliers are probed in the amplitude domain and are digitized by FPGA-based readout boards in triggerless mode. In this contribution we report on the first two- and three-photon positronium images and tests of CPT symmetry in positronium decays