Identification of deleterious germline CHEK2 mutations and their association with breast and ovarian cancer

Germline mutations in checkpoint kinase 2 (CHEK2), a multiple cancer-predisposing gene, increase breast cancer (BC) risk; however, risk estimates differ substantially in published studies. We analyzed germline CHEK2 variants in 1,928 high-risk Czech breast/ovarian cancer (BC/OC) patients and 3,360 population-matched controls (PMCs). For a functional classification of VUS, we developed a complementation assay in human nontransformed RPE1-CHEK2-knockout cells quantifying CHK2-specific phosphorylation of endogenous protein KAP1. We identified 10 truncations in 46 (2.39%) patients and in 11 (0.33%) PMC (p = 1.1 × 10⁻¹⁴). Two types of large intragenic rearrangements (LGR) were found in 20/46 mutation carriers. Truncations significantly increased unilateral BC risk (OR = 7.94; 95%CI 3.90–17.47; p = 1.1 × 10⁻¹⁴) and were more frequent in patients with bilateral BC (4/149; 2.68%; p = 0.003), double primary BC/OC (3/79; 3.80%; p = 0.004), male BC (3/48; 6.25%; p = 8.6 × 10⁻⁴), but not with OC (3/354; 0.85%; p = 0.14). Additionally, we found 26 missense VUS in 88 (4.56%) patients and 131 (3.90%) PMC (p = 0.22). Using our functional assay, 11 variants identified in 15 (0.78%) patients and 6 (0.18%) PMC were scored deleterious (p = 0.002). Frequencies of functionally intermediate and neutral variants did not differ between patients and PMC. Functionally deleterious CHEK2 missense variants significantly increased BC risk (OR = 3.90; 95%CI 1.24–13.35; p = 0.009) and marginally OC risk (OR = 4.77; 95%CI 0.77–22.47; p = 0.047); however, carriers low frequency will require evaluation in larger studies. Our study highlights importance of LGR detection for CHEK2 analysis, careful consideration of ethnicity in both cases and controls for risk estimates, and demonstrates promising potential of newly developed human nontransformed cell line assay for functional CHEK2 VUS classification.     

Influence of Al/Ti Ratio and Ta Concentration on the As-Cast Microstructure,Phase Composition,and Phase Transformation Temperatures of Lost-Wax Ni-Based Superalloy Castings

The as-cast microstructure, alloying element segregation, solidification behavior, and thermal stability of model superalloys based on Inconel 740 with various Al/Ti ratios (0.7, 1.5, 3.4) and Ta (2.0, 3.0, 4.0 wt%) concentrations were investigated via ThermoCalc simulations, scanning and transmission electron microscopy, energy-dispersive X-ray spectroscopy, dilatometry, and differential scanning calorimetry. The solidification of the superalloys began with the formation of primary γ dendrites, followed by MC carbides. The type of subsequently formed phases depended on the superalloys’ initial Al/Ti ratio and Ta concentration. The results obtained from solidification simulations were compared to the obtained microstructures. For all castings, the dendritic regions consisted of fine γ′ precipitates, with their size mainly depending on the initial Al/Ti ratio, whereas in the interdendritic spaces, (Nb, Ta, Ti)C carbides and Nb-rich Laves phase precipitates were present. In high Al/Ti ratio superalloys, β-NiAl precipitates, strengthened by η and α-Cr phases, were observed. Based on dilatometric results, the dissolution of γ′ precipitates was accompanied by a substantial increase in the coefficient of thermal expansion. The end of the dilatation effect took place around the γ′ solvus temperature, as determined via calorimetry. Moreover, the bulk solidus temperature was preceded by the dissolution of the Laves phase, which may be accompanied by local melting.     

LncRNA PVT1 is increased in renal cell carcinoma and affects viability and migration in vitro

BackgroundRenal cell carcinoma is difficult to diagnose and unpredictable in disease course and severity. There are no specific biomarkers for diagnosis and prognosis estimation feasible in clinical practice. Long non‐coding RNAs (lncRNAs) have emerged as potent regulators of gene expression in recent years. Aside from their cellular role, their expression patterns could be used as a biomarker of ongoing pathology.MethodsIn this work, we used next‐generation sequencing for global lncRNA expression profiling in tumor and non‐tumor tissue of RCC patients. The four candidate lncRNAs have been further validated on an independent cohort. PVT1, as the most promising lncRNA, has also been studied using functional in vitro tests.ResultsNext‐generation sequencing showed significant dysregulation of 1163 lncRNAs; among them top 20 dysregulated lncRNAs were AC061975.7, {"type":"entrez-nucleotide","attrs":{"text":"AC124017.1","term_id":"21327652","term_text":"AC124017.1"}}AC124017.1, {"type":"entrez-nucleotide","attrs":{"text":"AP000696.1","term_id":"6705905","term_text":"AP000696.1"}}AP000696.1, AC148477.4, LINC02437, GATA3‐AS, LINC01762, LINC01230, LINC01271, LINC01187, LINC00472, {"type":"entrez-nucleotide","attrs":{"text":"AC007849.1","term_id":"5091570","term_text":"AC007849.1"}}AC007849.1, LINC00982, LINC01543, {"type":"entrez-nucleotide","attrs":{"text":"AL031710.1","term_id":"3821014","term_text":"AL031710.1"}}AL031710.1, and {"type":"entrez-nucleotide","attrs":{"text":"AC019197.1","term_id":"6648325","term_text":"AC019197.1"}}AC019197.1 as down‐regulated lncRNAs; and SLC16A1‐AS1, PVT1, LINC0887, and LUCAT1 as up‐regulated lncRNAs. We observed statistically significant dysregulation of PVT1, LUCAT1, and LINC00982. Moreover, we studied the effect of artificial PVT1 decrease in renal cell line 786–0 and observed an effect on cell viability and migration.ConclusionOur results show not only the diagnostic but also the therapeutic potential of PVT1 in renal cell carcinoma.     

Psychodynamic day treatment program for borderline personality disorder: factors that predict outcome and dropout: An observational study

The objective of this study was to ascertain changes in symptoms of patients with borderline personality disorder undergoing psychodynamic day treatment with a duration of 9 months and the factors that predict clinical outcome or dropouts from the program.In an observational study, demographic characteristics (age, number of psychiatric hospitalizations, number of suicide attempts, current involvement in work or study activities), day doses of antipsychotic and antidepressant medication, psychiatric symptoms, and social functioning (Health of the Nation Outcome Scales), and symptoms of dissociation (Dissociative Experiences Scale) were assessed in patients at the beginning of treatment (N = 105). Further, psychiatric symptoms and social functioning were assessed at 3 stages: beginning of the program, end of the program, and 1-year follow-up. To study the differences between baseline values and values at the end of the treatment and follow-up values, the Wilcoxon signed-rank test was used. To discover baseline factors related to the effect of the treatment, Spearman correlation coefficients were calculated. To evaluate the differences between patients who completed the program (N = 67) and patients who dropped out (N = 38), differences in baseline factors between both groups were compared, using the Mann–Whitney test for independent samples.Improvement in symptoms (Health of the Nation Outcome Scales – version for external evaluators) at the end of the therapy (N = 67, P < .001) and at the 1-year follow-up (N = 46, P < .001) was found. Experience of an intimate relationship was positively related to clinical improvement at follow-up examinations (P < .001). Predictors of dropout included a higher number of psychiatric hospitalizations (P = .004), suicide attempts (P = .004), more severe pretreatment symptoms (P = .002), and symptoms of dissociation (P = .046).The results indicate that a psychodynamic day treatment is feasible for the treatment of less clinically disturbed patients with a history of intimate relationships. Patients with a higher number of previous psychiatric hospitalizations, more suicide attempts in the past, more severe pretreatment symptoms, and symptoms of dissociation are more likely not to complete the program.     

Morphological aspects of small-duct cholangiopathies: A minireview

The biliary system consists of intrahepatic and extrahepatic bile ducts lined by biliary epithelial cells (cholangiocytes). Bile ducts and cholangiocytes are affected by a variety of disorders called cholangiopathies, which differ in aetiology, pathogenesis, and morphology. Classification of cholangiopathies is complex and reflects pathogenic mechanisms (immune-mediated, genetic, drug- and toxin-induced, ischaemic, infectious, neoplastic), predominant morphological patterns of biliary injury (suppurative and non-suppurative cholangitis, cholangiopathy), and specific segments of the biliary tree affected by the disease process. While the involvement of large extrahepatic and intrahepatic bile ducts is typically visualised using radiology imaging, histopathological examination of liver tissue obtained by percutaneous liver biopsy still plays an important role in the diagnosis of cholangiopathies affecting the small intrahepatic bile ducts. To increase the diagnostic yield of a liver biopsy and determine the optimal therapeutic approach, the referring clinician is tasked with interpreting the results of histopathological examination. This requires knowledge and understanding of basic morphological patterns of hepatobiliary injury and an ability to correlate microscopic findings with results obtained by imaging and laboratory methods. This minireview describes the morphological aspects of small-duct cholangiopathies pertaining to the diagnostic process.     

Safety and efficacy of one and two booster doses of SARS-CoV-2 mRNA vaccines in kidney transplant recipients: A randomized clinical trial

Background

Kidney transplant recipients are at risk for a severe course of COVID-19 with a high mortality rate. A considerable number of patients remains without a satisfactory serological response after the baseline and adjuvant SARS-CoV-2 vaccination schedule.

Methods

In this prospective, randomized study, we evaluated the efficacy and safety of one and two booster doses of mRNA vaccines (either mRNA-1273 or BNT162b2) in 125 COVID-19 naive, adult kidney transplant recipients who showed an insufficient humoral response (SARS-CoV-2 IgG <10 AU/ml) to the previous 2-dose vaccination schedule. The primary outcome was the difference in the rate of a positive antibody response (SARS-CoV-2 IgG ≥10 AU/ml) between one and two booster doses at 1 month after the final booster dose.

Results

A positive humoral response was observed in 36 (62%) patients receiving two booster doses and in 28 (44%) patients receiving one booster dose (odds ratio [OR], 2.10, 95% confidence interval [CI], 1.02–4.34, p = .043). Moreover, median SARS-CoV-2 IgG levels were higher with two booster doses (p = .009). The number of patients with positive virus neutralizing antibody (VNA) levels was numerically higher with two booster doses compared to one booster dose, but without statistical significance (66% vs. 50%, p = .084). There was no significant difference in positive seroconversions rate and antibody levels between mRNA-1273 and BNT162b2.

Conclusion

A higher number of kidney transplant recipients achieved a positive antibody response after two booster doses compared to one booster dose.

     

Elevated erythroferrone distinguishes erythrocytosis with inherited defects in oxygen-sensing pathway from primary familial and congenital polycythaemia

Congenital erythrocytoses represent a heterogenous group of rare defects of erythropoiesis characterized by elevated erythrocyte mass. We performed molecular-genetic analysis of 21 Czech patients with congenital erythrocytosis and assessed the mutual link between chronic erythrocyte overproduction and iron homoeostasis. Causative mutations in erythropoietin receptor (EPOR), hypoxia-inducible factor 2 alpha (HIF2A) or Von Hippel–Lindau (VHL) genes were detected in nine patients, including a novel p.A421Cfs*4 EPOR and a homozygous intronic c.340+770T>C VHL mutation. The association and possible cooperation of five identified missense germline EPOR or Janus kinase 2 (JAK2) variants with other genetic/non-genetic factors in erythrocytosis manifestation may involve variants of Piezo-type mechanosensitive ion channel component 1 (PIEZO1) or Ten-eleven translocation 2 (TET2), but this requires further research. In two families, hepcidin levels appeared to prevent or promote phenotypic expression of the disease. No major contribution of heterozygous haemochromatosis gene (HFE) mutations to the erythrocytic phenotype or hepcidin levels was observed in our cohort. VHL- and HIF2A-mutant erythrocytosis showed increased erythroferrone and suppressed hepcidin, whereas no overproduction of erythroferrone was detected in other patients regardless of molecular defect, age or therapy. Understanding the interplay between iron metabolism and erythropoiesis in different subgroups of congenital erythrocytosis may improve current treatment options.     

Recent advances in dopamine D2 receptor ligands in the treatment of neuropsychiatric disorders

Dopamine is a biologically active amine synthesized in the central and peripheral nervous system. This biogenic monoamine acts by activating five types of dopamine receptors (D_1–5Rs), which belong to the G protein-coupled receptor family. Antagonists and partial agonists of D₂Rs are used to treat schizophrenia, Parkinson's disease, depression, and anxiety. The typical pharmacophore with high D₂R affinity comprises four main areas, namely aromatic moiety, cyclic amine, central linker and aromatic/heteroaromatic lipophilic fragment. From the literature reviewed herein, we can conclude that 4-(2,3-dichlorophenyl), 4-(2-methoxyphenyl)-, 4-(benzo[b]thiophen-4-yl)-1-substituted piperazine, and 4-(6-fluorobenzo[d]isoxazol-3-yl)piperidine moieties are critical for high D₂R affinity. Four to six atoms chains are optimal for D₂R affinity with 4-butoxyl as the most pronounced one. The bicyclic aromatic/heteroaromatic systems are most frequently occurring as lipophilic appendages to retain high D₂R affinity. In this review, we provide a thorough overview of the therapeutic potential of D₂R modulators in the treatment of the aforementioned disorders. In addition, this review summarizes current knowledge about these diseases, with a focus on the dopaminergic pathway underlying these pathologies. Major attention is paid to the structure, function, and pharmacology of novel D₂R ligands, which have been developed in the last decade (2010–2021), and belong to the 1,4-disubstituted aromatic cyclic amine group. Due to the abundance of data, allosteric D₂R ligands and D₂R modulators from patents are not discussed in this review.