Influence of maternal cholesterol-enriched diet on chemical composition of teeth enamel in offspring of mice.

OBJECTIVE: Aim: To determine the chemical composition of the tooth enamel of two-day-old mice from hypercholesterolemic mothers by energy dispersive X-ray spectroscopy. PATIENTS AND METHODS: Materials and Methods: Forty mature female mice were randomly assigned (n = 20/group) to either a standard chow vivarium diet (control group) or a cholesterol-enriched chow diet (experimental group). After fertilization, pregnancy and birth, on postnatal day 2, the incisor segments of 6 pups form each group were used for energy dispersive X-ray spectroscopy. RESULTS: Results: Influence of maternal hypercholesterolemic diet on tooth development and mineralization was examined, which revealed changes in enamel chemical composition. First, the results indicate the presence of seven elements (Na, Cl, Ca, P, Mg, S, Fe) in the enamel of both the hypercholesterolemic and normal offspring, but the content of element Ca2+ decreased, the content of elements P5+, Na+, Cl- tended to increase in pups from hypercholesterolemic mice. Second, the initial level of mineralization according to the atomic (%) Ca / P in hypercholesterolemic pups ratio was 1.26, comparing with normal pups where level of mineralization was 1.34. Taking into account that irreversible changes in the structure of the enamel were observed when the Ca / P ratio was below 1.33, we can suggest that the eruption of teeth with an imperfect structure could be because of maternal hypercholesterolemic diet. CONCLUSION: Conclusions: Results of this study suggest that hypercholesterolemic diet during gestation and lactation leads to altered enamel mineralization in mice because of changes in chemical composition and may link to the early childhood caries.

Upregulation of ATP-Sensitive Potassium Channels as the Potential Mechanism of Cardioprotection and Vasorelaxation Under the Action of Pyridoxal-5-Phosphate in Old Rats.

Background: The aging process is accompanied by the weakening of the protective systems of the organism, in particular by the decrease in the expression of ATP-sensitive potassium (KATP) channels and in the synthesis of H2S. The aim of our work was to investigate the role of KATP channels in the cardioprotection induced by pyridoxal-5-phosphate (PLP) in aging. Methods: Experiments were performed on adult and old (aged 24 months) male Wistar rats, which were divided into 3 groups: adults, old, and old PLP-treated rats. PLP was administered orally once a day for 14 days at a dose of 0.7 mg/kg. The levels of mRNA expression of subunits KATP channels were determined by reverse transcription and real-time polymerase chain reaction analysis. Protein expression levels were determined by the Western blot. Cardiac tissue morphology was determined using transverse 6 µm deparaffinized sections stained with picrosirius red staining. Vasorelaxation responses of isolated aortic rings and the function of Langendorff-perfused isolated hearts during ischemia-reperfusion, H2S levels, and markers of oxidative stress were also studied. Results: Administration of PLP to old rats reduces cardiac fibrosis and improves cardiac function during ischemia-reperfusion and vasorelaxation responses to KATP channels opening. At the same time, there was a significant increase in mRNA and protein expression of SUR2 and Kir6.1 subunits of KATP channels, H2S production, and reduced markers of oxidative stress. The specific KATP channel inhibitor-glibenclamide prevented the enhancement of vasodilator responses and anti-ischemic protection in PLP-treated animals. Conclusions: We suggest that this potential therapeutic effect of PLP in old animals may be a result of increased expression of KATP channels and H2S production.

INFLUENCE OF CHOLESTEROL ENRICHED DIET ON GENES EXPRESSION ENCODING BONE MORPHOGENETIC PROTEIN-2 AND OSTEOCALCIN IN MOUSE MANDIBLE.

OBJECTIVE: The aim: To evaluate the mRNA expression of the key regulators of osteogenesis - osteocalcin and BMP-2 in the mouse embryos mandible (17th day of pregnancy) which were borne by females on high-cholesterol diet for 30 days before fertilization and throughout pregnancy. PATIENTS AND METHODS: Materials and methods: Experimental hypercholesterolemia (2%) was simulated by adding Cholesterol to the diet for 60 days. In experiment were used 40 mature female white mice that were randomly divided to control and experimental groups. The control group were fed with standard chow diet, the experimental group with diet with cholesterol enriched diet (with addition of 2 grams of Cholesterol per 100 grams of standard chow). The mandibles of mouse embryos (E-17) were examined by using molecular genetic methods. RESULTS: Results: In control group the relative level of BMP-2 mRNA / actin mRNA was 27.0«2.82, the relative level of and osteocalcin mRNA / actin mRNA was 30.5«6,28. In the jaws of animals in the experimental group with cholesterol enriched diet, the expression relative level of BMP-2 was 30.9«5.81 that is by 14,4% higher than in control group. Therefore, the expression level of оsteocalcin, on the contrary, decreased by 22.3% and was 23.7+5.31. CONCLUSION: Conclusions: Our study report influence of the cholesterol enriched diet (2%) on mRNA expression of BMP-2 and osteocalcin encoding genes. The embryos from mouse on cholesterol enriched diet (2%) had increased level of BMP-2 gene expression, however significantly decreased level of osteocalcin gene expression.

The performance and limitations of PCA3, TMPRSS2:ERG, HOXC6 and DLX1 urinary markers combined in the improvement of prostate cancer diagnostics.

BACKGROUND: Prostate cancer (PCa) is the second most commonly diagnosed cancer in men. To date, the role of the combined application of long non-coding RNAs (PCA3, DLX1, HOXC6, TMPRSS2:ERG) for obtaining the most accurate method of detection of PCa has not yet been comprehensively investigated. METHODS: In total 240 persons were included in the retrospective study. Among them were 150 patients with confirmed PCa, 30 patients with benign prostatic hyperplasia, 30 patients with active chronic prostatitis and 30 healthy volunteers. In all patients, the urine samples were collected prior to biopsy or treatment. Polymerase chain reaction with reverse transcription was performed to detect the expression level of PCA3, HOXC6, DLX1 and the presence of the TMPRSS2:ERG transcript. RESULTS: PCA3 was detected in urine samples in all cases. Using a PCA3 score of 56 allowed the differentiation between PCa and all other cases with a sensitivity of 61% and specificity of 96% (p < 0.001) while a PCA3 score threshold value of 50 resulted in a differentiation between clinically significant PCa (ISUP grades 2-5) and all other cases with a sensitivity of 93% and specificity of 93% (p < 0.001). The TMPRSS2:ERG expression in urine was detected exclusively in the group of patients with PCa and only in 16% of all cases. CONCLUSIONS: PCA3 score detected in urine demonstrated moderate sensitivity and good specificity in differentiation between PCa and non-PCa and high sensitivity and specificity in differentiation between clinically significant PCa and non-PCa.

Pharmacophore modeling, docking and molecular dynamics simulation for identification of novel human protein kinase C beta (PKCß) inhibitors.

Protein kinase Cß (PKCß) is considered as an attractive molecular target for the treatment of COVID-19-related acute respiratory distress syndrome (ARDS). Several classes of inhibitors have been already identified. In this article, we developed and validated ligand-based PKCß pharmacophore models based on the chemical structures of the known inhibitors. The most accurate pharmacophore model, which correctly predicted more than 70% active compounds of test set, included three aromatic pharmacophore features without vectors, one hydrogen bond acceptor pharmacophore feature, one hydrophobic pharmacophore feature and 158 excluded volumes. This pharmacophore model was used for virtual screening of compound collection in order to identify novel potent PKCß inhibitors. Also, molecular docking of compound collection was performed and 28 compounds which were selected simultaneously by two approaches as top-scored were proposed for further biological research. Supplementary Information: The online version contains supplementary material available at 10.1007/s11224-022-02075-y.

SINGLE NUCLEOTIDE POLYMORPHISM RS4696480 OF TLR2 GENE ASSOCIATES WITH SEVERITY OF ATOPIC DERMATITIS IN CHILDREN, BUT NOT WITH IGE SENSITIZATION TO MALASSEZIA.

OBJECTIVE: The aim: Malassezia has been linked to atopic dermatitis, and TLRs are suggested to mediate influence of Malassezia spp on human cells. The aim of the study was to examine if TLR2 rs4696480 polymorphism predisposes to atopic dermatitis, IgE sensitization to Malassezia or to severe phenotype of atopic dermatitis. PATIENTS AND METHODS: Materials and methods: The study included 103 patients with eczema and 84 healthy children. Specific IgE against Malassezia mix (m227) was analyzed in 47 patients using immunochemiluminescent method on the ImmunoCAP 100 (Thermo Fisher Scientific Inc., Phadia, Sweden). Genotyping for TLR2 rs4696480 was performed by using Real-time PCR. RESULTS: Results: Increased IgE to Malassezia spp. was observed in 34,3 % of children with eczema. Higher Malassezia spp.-specific IgE titre positively correlated with duration of atopic dermatitis and a higher total IgE. There were no difference in allele distribution among patients and control group (OR=1.096 (0.549- 2.191) for AT, OR=0.946 (0.430- 2.078) for TT, р > 0,05). TLR2 polymorphism rs4696480 was not associated with Malassezia spp.-sIgE. AA-genotype was significantly more frequent among patients with severe and moderate-to-severe AD (OR=6.395 (1.240-32.991). CONCLUSION: Conclusions: AA variant of TLR2 rs4696480 polymorphism predisposes to severe phenotype of AD.

Response of Circulating Inflammatory Markers to Intermittent Hypoxia-Hyperoxia Training in Healthy Elderly People and Patients with Mild Cognitive Impairment.

Intermittent hypoxia-hyperoxia training (IHHT) is a non-pharmacological therapeutic modality for management of some chronic- and age-related pathologies, such as Alzheimer's disease (AD). Our previous studies demonstrated significant improvement of cognitive function after IHHT in the patients with mild cognitive impairment (MCI). The present study further investigated the effects of IHHT on pro-inflammatory factors in healthy elderly individuals and patients with early signs of AD. Twenty-nine subjects (13 healthy subjects without signs of cognitive impairment syndrome and 16 patients diagnosed with MCI; age 52 to 76 years) were divided into four groups: Healthy+Sham (n = 7), Healthy+IHHT (n = 6), MCI+Sham (n = 6), and MCI+IHHT (n = 10). IHHT was carried out 5 days per week for 3 weeks (total 15 sessions), and each daily session included 4 cycles of 5-min hypoxia (12% FIO2) and 3-min hyperoxia (33% FIO2). Decline in cognitive function indices was observed initially in both MCI+Sham and MCI+IHHT groups. The sham training did not alter any of the parameters, whereas IHHT resulted in improvement in latency of cognitive evoked potentials, along with elevation in APP110, GDF15 expression, and MMP9 activity in both healthy subjects and those with MCI. Increased MMP2 activity, HMGB1, and P-selectin expression and decreased NETs formation and Aß expression were also observed in the MCI+IHHT group. There was a negative correlation between MoCA score and the plasma GDF15 expression (R = −0.5799, p < 0.05) before the initiation of IHHT. The enhanced expression of GDF15 was also associated with longer latency of the event-related potentials P330 and N200 (R = 0.6263, p < 0.05 and R = 0.5715, p < 0.05, respectively). In conclusion, IHHT upregulated circulating levels of some inflammatory markers, which may represent potential triggers for cellular adaptive reprogramming, leading to therapeutic effects against cognitive dysfunction and neuropathological changes during progression of AD. Further investigation is needed to clarify if there is a causative relationship between the improved cognitive function and the elevated inflammatory markers following IHHT.

THE EFFECT OF DIET ENRICHED WITH PYROPHOSPHATE (E450) ON MORPHOLOGICAL CHANGES OF TOOTH GERMS OF MOUSE EMBRYOS.

OBJECTIVE: The aim: To reveal the effect of pyrophosphates on the tooth germ structure in the mandible of embryos (17th day of pregnancy) gestated by females, kept on a pyrophosphate-rich diet since 30 days before fertilization to gestation. PATIENTS AND METHODS: Materials and methods: The effect of food supplements was studied in «Overload phosphates model¼. Experiments were carried out on white nonlinear outbred mice with mass 25-28g (n= 40). The females from the control group were fed with standard rodent food, whereas the experimental females were fed with pyrophosphate-enriched food. The material for the morphological study were the mandible of 17-day-old mouse embryos (E-17), which were examined under a microscope with subsequent photofixation. RESULTS: Results: The examination of the mandible of 17-day-old mouse embryos, gestated by females on a pyrophosphate-rich diet, showed morphological changes in tooth germs at the dental follicle development stage. CONCLUSION: Conclusions: The experimentation revealed that the pyrophosphate excessive intake during dental follicle development leads to early dentinogenesis and oppression of ectodermal structures of tooth germs.

Cardiac dysfunction in spontaneously hypertensive old rats is associated with a significant decrease of SUR2 expression.

ATP-sensitive potassium (KATP) channels are participants of mechanisms of pathological myocardial remodeling containment. The aim of our work was to find the association of changes in the expression of Kir6.1, Kir6.2, SUR1, and SUR2 subunits of KATP channels with changes in heart function and structure during aging under conditions of the constant increase of vascular pressure. The experiments were carried out on young and old spontaneously hypertensive rats (SHR) and Wistar rats. The expression levels of KATP channels subunits were determined using reverse transcription and quantitative PCR. It is shown that the mRNA expression level of Kir6.1 in young SHR rats is significantly lower (6.3-fold, p = 0.035) than that of young Wistar rats that may be one of the causes of arterial hypertension in SHR. At the same time, mRNA expression of both Kir6.1 and Kir6.2 in old SHR rats was significantly higher (6.8-fold, p = 0.003, and 5.9-fold, p = 0.006, respectively) than in young hypertensive animals. In both groups of old animals, SUR2 expression was significantly reduced compared to young animals, in Wistar rats at 3.87-fold (p = 0.028) and in SHR rats at 48.2-fold (p = 0.033). Changes in SUR1 expression were not significant. Thus, significant changes in the cardiovascular system, including impaired function and structure of the heart in old SHR rats, were associated with a significant decrease in SUR2 expression that may be one of the mechanisms of heart failure decompensation. Therefore, it can be assumed that increased expression of SUR2 may be one of the protective mechanisms against pathological myocardial remodeling.

Genotype Associations with the Different Phenotypes of Atopic Dermatitis in Children.

This study deals with detecting the associations of atopic dermatitis' (AD) phenotypes in children: alone or combined with seasonal allergic rhino-conjunctivitis (SARC) and/or perennial allergic rhinitis (PAR), and/or with bronchial asthma (BA) with single nucleotide polymorphisms (SNP) of filaggrin (FLG), thymic stromal lymphopoietin (TSLP) and orsomucoid-like-1 protein 3 (ORMDL3) genes. Male and female pediatric patients aged from 3 to 18 years old were recruited into the main (AD in different combinations with SARC, PAR, BA) and control groups (disorders of digestives system, neither clinical nor laboratory signs of atopy). Patients were genotyped for SNP of rs_7927894 FLG, rs_11466749 TSLP, rs_7216389 ORMDL3 variants. Statistically significant associations of the increased risk were detected of AD combined with SARC and/or PAR and AD combined with BA (possibly, SARC and/or PAR) with C/T rs_7927894 FLG and T/T rs_7216389 ORMDL3 genotypes. Genotype C/C rs_7927894 FLG significantly decreases the risk of AD combined with SARC and/or PAR by 2.56 fold. Several genotypes' associations had a trend to significance: C/C rs_7216389 ORMDL3 decreases and C/T rs_7216389 ORMDL3 increases the risk for developing AD alone phenotype; A/G rs_11466749 TSLP decreases the risk of AD combined with BA (possibly, SARC and/or PAR) phenotype development.

THE EFFECT OF DIET ENRICHED WITH PYROPHOSPHATE (E450) ON EXPRESSION OF GENES ENCODING BONE MORPHOGENETIC PROTEIN AND OSTEOCALCIN IN MOUSE EMBRYONIC MANDIBLE TISSUES.

OBJECTIVE: The aim: Of our study was to measure the mRNA expression of the investigated odontogenesis factors in mandible tissue of mouse embryos (17th day of pregnancy) gestated by females, kept on a E450 rich diet since 30 days before fertilization to gestation. PATIENTS AND METHODS: Materials and methods: The effect of food supplements was studied in «Overload phosphates model¼. Experiments were carried out on white nonlinear outbred mice with mass 25-28g (n=40). The females from the control group were fed with standard rodent food, whereas the experimental females were fed with pyrophosphate-enriched food. The materials, used for the molecular genetic study, were the lower jaws of 17-days old mouse embryos (E-17). RESULTS: Results: The investigated BMP2 and osteocalcin genes are expressed at approximately the same level. Pyrophosphate-rich diet does not alter BMP2 gene expression, but it significantly increases the expression of osteocalcin. CONCLUSION: Conclusions: The present study is the first one to describe the impact of the pyrophosphate-rich diet on mRNA expression of key osteogenesis regulators - osteocalcin and BMP2.

Relationship between mir-126 expression in children with psoriasis, disease progression and therapeutic response.

This study aimed to investigate the expression level of miR-126 in children with psoriasis in the epidermis affected by psoriasis and intact buccal epithelium, establish the impact on the characteristics of the course of psoriasis and the results of therapy in children with psoriasis of initial expression levels of miR-126. miR-126 expression levels in psoriatic keratinocytes and buccal epithelium were determined in 54 children with psoriasis on the severity of psoriasis, treatment efficacy. miR-126 levels in the buccal epithelium in children with psoriasis were reduced compared to healthy children (AUC=0.776±0.048, p<0.001). There were no discrepancies between miR-126 expression levels in psoriatic keratinocytes and buccal epithelium (p=0.097). There are statistically significant discrepancies between miR-126 expression levels in the psoriatic epidermis depending on the clinical form of psoriasis (AUC=0.637±0.056; p=0.014) and severity according to BSA (AUC=0.634±0.063; p=0.034). Depending on the miR-126 level in the buccal epithelium, the response to treatment (PASI<75) in children with high miR-126 is worse than in children with expected miR-126 levels (OR 2.79; 95%; CI: 1.19 - 6.51). Treatment failures were observed in children with high levels of miR-126 in the buccal epithelium compared to miR-126 in the psoriatic epidermis: children aged 12/13 to 17 years (OR 2.44; 95% CI: 1.02 - 5.85), children with PGA=4 (OR 3.16; 95% CI: 1.34 - 7.43). The location and level of miR-126 expression affects the course of psoriasis and the outcome of treatment. High levels of miR-126 in psoriatic keratinocytes lead to manifestations of plaque psoriasis with a course of moderate to severe forms. Initial miR-126 levels in the buccal epithelium in children with psoriasis are a prognostic criterion for response to therapy and can be used as a marker for prescribing systemic treatment.

PPARGC1A gene polymorphism is associated with exercise-induced fat loss.

Obesity is a widespread problem within modern society, serving to increase the risk of cardiovascular, metabolic, and neurodegenerative disorders. Peroxisome proliferator-activated receptor gamma (PPARγ) and PPARγ coactivator 1 α (PGC1α) play a key role in the regulation of cellular energy metabolism and is implicated in the pathology of these diseases. This study examined the association between polymorphisms of the PPARG and PPARGC1A genes and individual variability in weight loss in response to physical activity intervention. 39 obese Ukrainian women (44.4 ± 7.5 years, BMI > 30.0 kg/m2) undertook a 3-month fitness program whilst following a hypocaloric diet (~ 1500 cal). Anthropometric and biochemical measurements took place before and after the program. Single nucleotide polymorphisms within or near PPARG (n = 94) and PPARGC1A (n = 138) were identified and expression of PPARG mRNA was measured via reverse transcription and amplification. The association between DNA polymorphisms and exercise-induced weight loss, initial body mass, biochemistry and PPARG expression was determined using one-way analysis of variance (ANOVA). The present intervention induced significant fat loss in all participants (total fat: 40.3 ± 5.3 vs 36.4 ± 5.7%; P < 0.00001). Only one polymorphism (rs17650401 C/T) within the PPARGC1A gene was found to be associated with fat loss efficiency after correction for multiple testing, with T allele carriers showing the greatest reduction in body fat percentage (2.5-fold; P = 0.00013) compared to non-carriers. PPARGC1A (rs17650401) is associated with fat loss efficiency of the fitness program in obese women. Further studies are warranted to test whether this variation is associated with fat oxidation.

Regulation of microrna expression level by choleretic therapy in functional disorders of the gallbladder and оddi's sphincter in children.

OBJECTIVE: The alm: To study the effect of choleretic therapy on the level of microRNA expression in functional disorders of the gallbladder and Oddi's sphincter in children. PATIENTS AND METHODS: Materials and methods: Fifty patients with functional disorders of the gallbladder and Oddi's sphincter who received standard therapy in combination with ursodeoxycholic acid, 20 patients - standard therapy without ursodeoxycholic acid, and 20 healthy children were examined. The level of expression of microRNA-378f, microRNA-4311, microRNA-4714- 3p in the blood serum by the method of real-time polymerase chain reaction with reverse transcription according to the protocol TaqMan Gene Expression Assays was performed. RESULTS: Results: It was demonstrated that the activity profile of microRNA-4714-3p was significantly lower in those examined with functional disorders of the gallbladder and Oddi's sphincter than in practically healthy children (p<0.05). After standard therapy combined with ursodeoxycholic acid in children with functional disorders of the gallbladder and Oddi's sphincter, the level of expression of microRNA-378f is significantly higher than before therapy (5.23±0.70 SU and 2.02±0.57 SU respectively) (p<0.05). Against the background of standard therapy with the addition of ursodeoxycholic acid or without it, the expression profile of microRNA-4714-3p in the blood serum in children with functional disorders of the gallbladder and Oddi's sphincter significantly decreased (1.93±0.58 SU and 1,14±0,53 SU respectively) (p<0.05). CONCLUSION: Conclusions: Ursodeoxycholic acid in children with functional disorders of the gallbladder and Oddi's sphincter affects the activity of generation of gene regulators of the cellular mechanisms of microRNA-378f and microRNA-4714-3p.

Bioethical problems arising in the study of single-nucleotide gene polymorphisms of occupational diseases.

In Ukraine, about 3 million people work in hazardous and dangerous conditions. The study of hereditary specificity in development of occupational diseases is being actively conducted through molecular genetic analysis of single-nucleotide gene polymorphisms. While studying single-nucleotide gene polymorphisms of occupational diseases, many complicated bioethical questions arise regarding the confidentiality of personal data, the choice between the profession chosen and the risk to one's own health. Complicated bioethical issues that arise when studying single-nucleotide gene polymorphisms of occupational diseases need to be actively discussed, not only by physicians, occupational pathologists, employers, scientists, but also by politicians and lawyers, taking into account ethical and social norms and implications.

Expressions of microRNA-29a and microRNA-34a in pleomorphic adenomas of salivary glands.

BACKGROUND: The most common type of salivary gland tumor is pleomorphic adenoma. The genetic area of focus in the diagnosis of salivary gland tumors is a study of the role of miRNA. METHODS: Twenty-two patients with pleomorphic adenomas of the salivary glands were used for the examinations. The histological typing of the salivary gland tumors was performed when using routine staining with hematoxylin and eosin, as well as with immunohistochemistry. The expressions of miR-34a and miR-29a were evaluated by using reverse transcription and the quantitative polymerase chain reaction in a real-time setting. In addition, the study also calculated the levels of expression of miR-29a and miR-34a in the venous blood. RESULTS: The majority of patients-15 (68.18%) and 22 (100.00%) had a positive response to human papillomavirus (HPV) and pleomorphic adenoma gene 1 (PLAG1), respectively. The conducted analyses of the expressions of miR-34a and miR-29a showed that the highest expression was observed in the salivary gland tissue adjacent to the tumor (1,052.02±367.20 and 111.93±56.97, versus 47.72±28.93 and 8.12±4.40 in the intact salivary gland tissue, respectively). CONCLUSIONS: There was a sufficiently high level of miR-34a and miR-29a expressions in the tissues of the tumor of pleomorphic adenomas of the salivary glands when compared with the intact salivary gland tissue.

Intermittent Hypoxia-Hyperoxia Training Improves Cognitive Function and Decreases Circulating Biomarkers of Alzheimer's Disease in Patients with Mild Cognitive Impairment: A Pilot Study.

Alzheimer's disease (AD) affects not only the central nervous system, but also peripheral blood cells including neutrophils and platelets, which actively participate in pathogenesis of AD through a vicious cycle between platelets aggregation and production of excessive amyloid beta (Aß). Platelets adhesion on amyloid plaques also increases the risk of cerebral microcirculation disorders. Moreover, activated platelets release soluble adhesion molecules that cause migration, adhesion/activation of neutrophils and formation of neutrophil extracellular traps (NETs), which may damage blood brain barrier and destroy brain parenchyma. The present study examined the effects of intermittent hypoxic-hyperoxic training (IHHT) on elderly patients with mild cognitive impairment (MCI), a precursor of AD. Twenty-one participants (age 51-74 years) were divided into three groups: Healthy Control (n = 7), MCI+Sham (n = 6), and MCI+IHHT (n = 8). IHHT was carried out five times per week for three weeks (total 15 sessions). Each IHHT session consisted of four cycles of 5-min hypoxia (12% FIO2) and 3-min hyperoxia (33% FIO2). Cognitive parameters, Aß and amyloid precursor protein (APP) expression, microRNA 29, and long non-coding RNA in isolated platelets as well as NETs in peripheral blood were investigated. We found an initial decline in cognitive function indices in both MCI+Sham and MCI+IHHT groups and significant correlations between cognitive test scores and the levels of circulating biomarkers of AD. Whereas sham training led to no change in these parameters, IHHT resulted in the improvement in cognitive test scores, along with significant increase in APP ratio and decrease in Aß expression and NETs formation one day after the end of three-week IHHT. Such effects on Aß expression and NETs formation remained more pronounced one month after IHHT. In conclusion, our results from this pilot study suggested a potential utility of IHHT as a new non-pharmacological therapy to improve cognitive function in pre-AD patients and slow down the development of AD.

Comorbidity of asthma and hypertension may be mediated by shared genetic dysregulation and drug side effects.

Asthma and hypertension are complex diseases coinciding more frequently than expected by chance. Unraveling the mechanisms of comorbidity of asthma and hypertension is necessary for choosing the most appropriate treatment plan for patients with this comorbidity. Since both diseases have a strong genetic component in this article we aimed to find and study genes simultaneously associated with asthma and hypertension. We identified 330 shared genes and found that they form six modules on the interaction network. A strong overlap between genes associated with asthma and hypertension was found on the level of eQTL regulated genes and between targets of drugs relevant for asthma and hypertension. This suggests that the phenomenon of comorbidity of asthma and hypertension may be explained by altered genetic regulation or result from drug side effects. In this work we also demonstrate that not only drug indications but also contraindications provide an important source of molecular evidence helpful to uncover disease mechanisms. These findings give a clue to the possible mechanisms of comorbidity and highlight the direction for future research.

MicroRNA-15a tissue expression is a prognostic marker for survival in patients with clear cell renal cell carcinoma.

None of the currently investigated molecular markers demonstrated sufficient accuracy in prognostication of the renal cell carcinoma (RCC) oncologic outcomes; thus, none of them has been recommended for the application in the routine clinical practice. The role of miR-15a as a potential prognostic marker for RCC is still not unveiled. The aim of our study was to assess the expression of miR-15a in tumor tissues of the patients with RCC and to evaluate the possibility of its usage as a prognostic molecular biomarker of this disease. The retrospective included 64 adult patients with clear cell RCC (ccRCC) in whom radical or partial nephrectomy was conducted. After deparaffinization of formalin-fixed paraffin-embedded (FFPE) ccRCC specimens, the tissue expression of miR-15a was measured using the reverse transcription and quantitative polymerase chain reaction in the real time. For the reference, the expression of miR-15a was estimated in 15 FFPE tissue specimens of the normal renal parenchyma. Survival analysis involved all cases of non-metastatic RCCs (n = 57). Five-year cancer-specific survival (CSS) was estimated by means of the Kaplan-Meier method and was calculated from the date of surgery to the date of death. Patients with the RCC were characterized by significantly upregulated tumor tissue mean levels of miR-15a compared to the healthy controls: 0.10 ± 2.62 relative units (RU) versus 4.84E - 03 ± 3.11E - 03 RU (p < 0.001). Overexpression of miR-15a was strongly associated with poor histologic prognostic features of ccRCC. Poorly differentiated tumors tend to have more pronounced upregulation of miR-15a compared to highly differentiated lesions: Mean expression values were 4.57 ± 3.19 RU for Fuhrman grade 4 versus 0.02 ± 0.01 RU for Fuhrman grade 1 (p < 0.001). The metastatic involvement of the regional lymphatic nodules (N +) was associated with significantly upregulated miRNA-15a in comparison with N - cases: Mean expression values were 4.92 ± 2.80 RU versus 1.10 ± 2.29 RU, respectively (p < 0.001). In patients with miR-15a expression in RCC tissues ≤ 0.10 RU, mean 5-year CSS was significantly longer compared to patients with expression levels above this threshold: 92.31% (mean duration of survival-59.88 ± 0.12 months) versus 54.8% (mean duration of survival-49.74 ± 2.16 months), respectively (p < 0.001). The tissue expression of miR-15a could be used as a potential prognostic molecular biomarker for conventional RCC.

Cholesterol enriched diet suppresses ATF6 and PERK and upregulates the IRE1 pathways of the unfolded protein response in spontaneously hypertensive rats: Relevance to pathophysiology of atherosclerosis in the setting of hypertension.

Many studies have been dedicated to hypertension and hypercholesterolemia, as they are the primary conditions that influence the unfolded protein response (UPR). However, the concurrent effects of these two factors are unknown. Our research used spontaneously hypertensive rats (SHR) fed a cholesterol enriched diet (CED) as model of atherosclerosis formation to discover what effect the simultaneous actions of hypertension and hypercholesterolemia have on the UPR. The combination of hypertension and consumption of a CED (not the CED alone) caused the formation of early atherosclerotic features. Both increased expression of the CCAAT-enhancer-binding protein (CHOP) and the insulin induced gene 1 (INSIG1), which is the target gene of the sterol regulatory element-binding protein 1-c (SREBP1-c), and decreased expression of the spliced x-box binding protein1 (sXBP1) mRNA were observed in the SHR fed a CED. Cholesterol overload strongly suppressed glucose regulated protein 78 (GRP78), glucose regulated protein 94 (GRP 94), and the expression of CHOP and INSIG1 mRNA in both normotensive and hypertensive rats. Unlike other UPR factors, the sXBP1 mRNA expression was strongly downregulated in SHR fed a normal diet but upregulated in those fed a CED. The changes to UPR in the SHR fed a CED were associated with improvement of the initially impaired heart function of the rats.