Intrahepatic expression of genes related to metabotropic receptors in chronic hepatitis

AIM: To screen for genes related to metabotropic receptors that might be involved in the development of chronic hepatitis. METHODS: Assessment of 20 genes associated with metabotropic receptors was performed in liver specimens obtained by punch biopsy from 12 patients with autoimmune and chronic hepatitis type B and C. For this purpose, a microarray with low integrity grade and with oligonucleotide DNA probes complementary to target transcripts was used. Evaluation of gene expression was performed in relation to transcript level, correlation between samples and grouping of clinical parameters used in chronic hepatitis assessment. Clinical markers of chronic hepatitis included alanine and aspartate aminotransferase, γ-glutamyltranspeptidase, alkaline phosphatase and cholinesterase activity, levels of iron ions, total cholesterol, triglycerides, albumin, glucose, hemoglobin, platelets, histological analysis of inflammatory and necrotic status, fibrosis according to METAVIR score, steatosis, as well as anthropometric body mass index, waist/hip index, percentage of adipose tissue and liver size in ultrasound examination. Gender, age, concomitant diseases and drugs were also taken into account. Validation of oligonucleotide microarray gene expression results was done with the use of quantitative real-time polymerase chain reaction (qRT-PCR). RESULTS: The highest (0.002 < P < 0.046) expression among genes encoding main components of metabotropic receptor pathways, such as the α subunit of G-coupled protein, phosphoinositol-dependent protein kinase or arrestin was comparable to that of angiotensinogen synthesized in the liver. Carcinogenesis suppressor genes, such as chemokine ligand 4, transcription factor early growth response protein 1 and lysophosphatidic acid receptor, were characterized by the lowest expression (0.002 < P < 0.046), while the factor potentially triggering hepatic cancer, transcription factor JUN-B, had a 20-fold higher expression. The correlation between expression of genes of protein kinases PDPK1, phosphoinositide 3-kinase and protein kinase A (Spearman’s coefficient range: 0.762-0.769) confirmed a functional link between these enzymes. Gender (P = 0.0046) and inflammation severity, measured by alanine aminotransferase activity (P = 0.035), were characterized by diverse metabotropic receptor gene expression patterns. The Pearson’s coefficient ranging from -0.35 to 0.99 from the results of qRT-PCR and microarray indicated that qRT-PCR had certain limitations as a validation tool for oligonucleotide microarray studies. CONCLUSION: A microarray-based analysis of hepatocyte metabotropic G-protein-related gene expression can reveal the molecular basis of chronic hepatitis

Long-term space missions’ effects on the human organism: what we do know and what requires further research

Space has always fascinated people. Many years have passed since the first spaceflight, and in addition to the enormous technological progress, the level of understanding of human physiology in space is also increasing. The presented paper aims to summarize the recent research findings on the influence of the space environment (microgravity, pressure differences, cosmic radiation, etc.) on the human body systems during short-term and long-term space missions. The review also presents the biggest challenges and problems that must be solved in order to extend safely the time of human stay in space. In the era of increasing engineering capabilities, plans to colonize other planets, and the growing interest in commercial space flights, the most topical issues of modern medicine seems to be understanding the effects of long-term stay in space, and finding solutions to minimize the harmful effects of the space environment on the human body

Cobalt protoporphyrin IX increases endogenous G-CSF and mobilizes HSC and granulocytes to the blood

Granulocyte colony-stimulating factor (G-CSF) is used in clinical practice to mobilize cells from the bone marrow to the blood; however, it is not always effective. We show that cobalt protoporphyrin IX (CoPP) increases plasma concentrations of G-CSF, IL-6, and MCP-1 in mice, triggering the mobilization of granulocytes and hematopoietic stem and progenitor cells (HSPC). Compared with recombinant G-CSF, CoPP mobilizes higher number of HSPC and mature granulocytes. In contrast to G-CSF, CoPP does not increase the number of circulating T cells. Transplantation of CoPP-mobilized peripheral blood mononuclear cells (PBMC) results in higher chimerism and faster hematopoietic reconstitution than transplantation of PBMC mobilized by G-CSF. Although CoPP is used to activate Nrf2/HO-1 axis, the observed effects are Nrf2/HO- 1 independent. Concluding, CoPP increases expression of mobilization- related cytokines and has superior mobilizing efficiency compared with recombinant G-CSF. This observation could lead to the development of new strategies for the treatment of neutropenia and HSPC transplantation

The I/D ACE gene polymorphism and antihypertensive efficacy of losartan in patients with primary hypertension

Wstęp Polimorfizm insercyjno/delecyjny (I/D) genu ACE kodującego konwertazę angiotensyny I jest najchętniej analizowanym i najlepiej przebadanym wariantem sekwencji w genomie. Jego istotne znaczenie czynnościowe sprawia, że jest częstym obiektem badań typu analizy związku w odniesieniu do praktycznie wszystkich chorób układu sercowo-naczyniowego i ich powikłań. Trwają badania dotyczące związku polimorfizmu I/D genu ACE z pierwotnym nadciśnieniem tętniczym oraz farmakogenetyką leków hipotensyjnych, w tym antagonistów receptora angiotensyny II (ARB). Celem niniejszej pracy była ocena związku pomiędzy polimorfizmem I/D genu ACE a działaniem hipotensyjnym losartanu (podawanego przez 8 tygodni) u chorych na pierwotne nadciśnienie tętnicze. Materiał i metody Do badania włączono 50 osób (40 mężczyzn i 10 kobiet) w wieku średnio 47 &plusmn; 8 lat, z nadciśnieniem tętniczym (stopień I-II wg ESH/ /ESC, 2007). Wszyscy chorzy przez 8 tygodni byli poddawani leczeniu antagonistą receptora angiotensyny II - losartanem w dawce 50 mg raz na dobę. W przypadku braku normalizacji ciśnienia (< 140/90 mm Hg) po 4 tygodniach zwiększano dawkę leku do 100 mg raz na dobę. U wszystkich osób włączonych do badania wykonywano pomiary ciśnienia tętniczego metodą tradycyjną (przed oraz po 4 i 8 tygodniach leczenia), całodobową rejestrację ciśnienia tętniczego (ABPM) i podstawowe badania biochemiczne przed i po 8 tygodniach leczenia; oznaczano też stężenia angiotensyny II w osoczu i aktywność reninową osocza (ARO) w warunkach podstawowych oraz po 8 tygodniach terapii. Genotypy I/D ACE określano metodą łańcuchowej reakcji polimerazy (PCR). Wyniki W badanej grupie chorych stwierdzono: 10 homozygot II (20%), 25 heterozygot ID (50%) oraz 15 homozygot DD (30%). Częstość występowania allelu I wynosiła 45%, a allelu D - 55%. Pomiędzy pacjentami o różnych genotypach nie stwierdzono istotnych różnic w zakresie wieku, płci, wskaźnika masy ciała, parametrów przemiany lipidowej oraz wyjściowych wartości skurczowego (SBP) i rozkurczowego (DBP) ciśnienia tętniczego. Po 8 tygodniach leczenia u wszystkich pacjentów stwierdzono istotne obniżenie zarówno SBP, jak i DBP w pomiarach tradycyjnych oraz ABPM. Dawkę losartanu zwiększono do 100 mg u 23 osób, w tym u 7 (70%) z genotypem II, 12 z genotypem ID (48%) i 4 z genotypem DD (27%). Po 8 tygodniach leczenia wartość DBP poniżej 90 mm Hg osiągnięto u wszystkich chorych z genotypem DD. Zarówno wartości SBP,jak i DBP po 8 tygodniach leczenia były istotnie niższe u osób z genotypem DD niż u pacjentów z allelem I (ID lub II). Wnioski Wyniki badania wskazują na związek pomiędzy polimorfizmem I/D genu ACE a działaniem hipotensyjnym wywieranym przez antagonistę receptora angiotensyny II - losartan. Nadciśnienie Tętnicze 2007, tom 11, nr 6, strony 498-504.Background The insertion/deletion (I/D) ACE gene polymorphism seems the most widely studied sequence variant of human genome. Although evidence implicates the linkage of ACE locus and the risk for arterial hypertension, there are confounding data regarding association of ACE gene polymorphism with blood pressure (BP) response to antihypertensive therapy in patients with essential hypertension. Therefore the aim of our study was to evaluate the association between I/D ACE polymorphism and antihypertensive efficacy of an angiotensin II receptor blocker - losartan. Material and methods Fifty patients (mean age 47 &plusmn; 8 years) with essential hypertension (stage I-II , ESH/ESC, 2007) were included into the study. The patients were treated with losartan starting from dose of 50 mg once daily. The dose could be increased up to 100 mg once daily if there was no normalization of blood pressure after 4 weeks of treatment. The ambulatory BP measurements (ABPM), clinic BP measurements as well as evaluation of plasma renin activity and serum angiotensin II concentration measurements were performed before and after 8 weeks of treatment. The ACE genotypes were identified by PCR method. Results The frequency distribution of ACE genotypes were as follows: 20% II homozygotes, 50% ID heterozygotes and 30% DD homozygotes. There were no significant differences in clinic characteristics and baseline BP levels among II, ID or DD patients. After 8 weeks of treatment significant decrease of BP levels both in clinic measurements and ABPM was noted regardless of ACE genotype. Losartan dose was titrated in 7 patients with II genotype (70%), 12 patients with ID genotype (48%) and in 4 patients with DD genotype (27%). Patients with DD genotype were characterized by lower systolic and diastolic BP levels after 8 weeks of treatment as compared with carriers of the I allele (ID or II). Conclusions Our results suggest the association between I/D ACE gene polymorphism and the hypotensive effect of losartan.Arterial Hypertension 2007, vol. 11, no 6, pages 498-504

The role of school functioning, physical activity, BMI, sex and age in building resilience among Ukrainian refugee children in Poland

Abstract The study aims to examine the relationship between school functioning, physical activity (PA), sex, Body Mass Index (BMI), age, and resilience in Ukrainian children who migrated to Poland due to the war. A cross-sectional study was conducted in 2022, focusing on 248 children aged 10–15 years. The findings suggest that school environment, including enjoyment of school and strong support from teachers, plays a significant role in building resilience in children. PA enhanced the resilience of girls, whereas a higher BMI negatively impacted it. A child-friendly school environment that encourages PA and provides social support could be a promising approach for the mental health of Ukrainian refugee children

Qualitative description of detachment forces for macromolecules

Dynamic force spectroscopy provides insight into the structure and dynamics of interacting molecules or surfaces at a molecular level by applying mechanical forces in a controllable way. The use of atomic force microscopy (AFM) in single-molecule pulling experiments allows studying forced escape, detachment, dissociation events associated with an increasing force field, and determining bonds’ strength. Here, we consider the rupture force spectra in a poly(ethyleneimine) (PEI) model system deposited on a silica surface displaying multiple dissociation events in contact with the AFM probe and analyze statistical properties of rupture forces derived from the multiple repeated experiments. We find that the obtained histograms are overdispersed. The most probable rupture force decreases with the increase of pulling velocity being of poor agreement with models based on Kramers theory describing the crossing event through a single energy barrier. The experimental data of multi-breakdown events are shown to be well fitted with the Weibull distribution, which stems from generally nonexponential molecular relaxation under the action of applied stress. We also provide a numerical study of a simple microscopic polymer chain model with multiple bonds with the surface, as well as a stochastic PEI model. The obtained results support our experimental and theoretical findings qualitatively

Arabidopsis ABA-activated kinase MAPKKK18 is regulated by protein phosphatase 2C ABI1 and the ubiquitin proteasome pathway

Phosphorylation and dephosphorylation events play an important role in the transmission of the abscisic acid signal. Although SnRK2 protein kinases and group A PP2C-type phosphatases constitute a core ABA pathway, little is known about the interplay between MAP kinases and protein phosphatases 2C in the regulation of ABA pathways. In this study, an effort was made to elucidate the role of MKKK18 in relation to ABA signaling and response. The MKKK18 knockout lines showed more vigorous root growth, decreased abaxial stomatal index, and increased stomatal aperture under normal growth conditions, compared to the control WT Col-0 line. In addition to transcriptional regulation of the MKKK18 promoter by ABA, we demonstrated by using in vitro and in vivo kinase assays that the kinase activity of MKKK18 was regulated by ABA and in response to H2O2 treatment. Analysis of the cellular localization of MKKK18 showed that the active kinase was targeted specifically to the nucleus. Notably, we identified ABI1 PP2C as a MKKK18-interacting protein, and demonstrated that ABI1 inhibited its activity. Using a cell-free degradation assay, we also established that MKKK18 was unstable and was degraded by the proteasome pathway. The rate of MKKK18 degradation was delayed in the ABI1 knockout line. Overall, we provide evidence that ABI1 regulates activity and promotes proteasomal degradation of MKKK18