Expression Profiles of Dopamine-Related Genes and miRNAs Regulating Their Expression in Breast Cancer.

This study aimed to assess the expression profile of messenger RNA (mRNA) and microRNA (miRNA) related to the dopaminergic system in five types of breast cancer in Polish women. Patients with five breast cancer subtypes were included in the study: luminal A (n = 130), luminal B (n = 196, including HER2-, n = 100; HER2+, n = 96), HER2+ (n = 36), and TNBC (n = 43); they underwent surgery, during which tumor tissue was removed along with a margin of healthy tissue (control material). The molecular analysis included a microarray profile of mRNAs and miRNAs associated with the dopaminergic system, a real-time polymerase chain reaction preceded by reverse transcription for selected genes, and determinations of their concentration using enzyme-linked immunosorbent assay (ELISA). The conducted statistical analysis showed that five mRNAs statistically significantly differentiated breast cancer sections regardless of subtype compared to control samples; these were dopamine receptor 2 (DRD2), dopamine receptor 3 (DRD3), dopamine receptor 25 (DRD5), transforming growth factor beta 2 (TGF-ß-2), and caveolin 2 (CAV2). The predicted analysis showed that hsa-miR-141-3p can regulate the expression of DRD2 and TGF-ß-2, whereas hsa-miR-4441 is potentially engaged in the expression regulation of DRD3 and DRD5. In addition, the expression pattern of DRD5 mRNA can also be regulated by has-miR-16-5p. The overexpression of DRD2 and DRD3, with concomitant silencing of DRD5 expression, confirms the presence of dopaminergic abnormalities in breast cancer patients. Moreover, these abnormalities may be the result of miR-141-3P, miR-16-5p, and miR-4441 activity, regulating proliferation or metastasis.

Impact of Nutritional Status of Patients with Head and Neck Squamous Cell Carcinoma on the Expression Profile of Ghrelin, Irisin, and Titin.

The goal of this paper was the evaluation of the changes in the expression profile of irisin, ghrelin, and titin in the carcinoma tissue and in the blood of patients with head and neck squamous cell carcinoma (HNSCC), including determining the profile of their expression in relation to patient nutrition. The study included 56 patients with diagnosed squamous cell carcinoma of HNSCC in the T3 and T4 stages of the disease. Healthy control tissue specimens were collected from an area 10 mm outside the histologically negative margin. In turn, the blood and serum from the control group came from healthy volunteers treated for non-oncologic reasons (n = 70). The molecular analysis allowed us to determine the profile of irisin, ghrelin, and titin methylation, evaluate their expression on the level of mRNA (quantitative Reverse Transcription Polymerase Chain Reaction; qRT-PCR) and protein (Enzyme-Linked Immunosorbent Assay Reaction; ELISA) in the carcinoma tissue and the margin of healthy tissue, as well as in serum of patients in the study and control groups. At the start of our observations, a Body Mass Index (BMI) < 18.5 was noted in 42 of the patients, while six months after the treatment a BMI < 18.5 was noted in 29 patients. We also noted a decrease in the expression of irisin, ghrelin, and titin both on the level of mRNA and protein, as well as a potential regulation of their expression via DNA methylation. There is no convincing evidence that the proteins assayed in the present work are specific with regard to HNSSC.

Influence of adalimumab on interleukin 12/23 signalling pathways in human keratinocytes treated with lipopolysaccharide A.

Introduction: The interleukin-12/23 (IL-12/23) signalling pathway plays an important role in the pathogenesis of psoriasis. In addition, even molecularly targeted therapy has been reported to lose adequate response to treatment. Aim: To determine the expression patterns of mRNAs and miRNAs related to IL-12/23 signalling pathways in the human keratinocyte culture exposed to liposaccharide A (LPS) and then adalimumab in comparison with untreated cells. Material and methods: Human, adult, low-Calcium, high-Temperature keratinocyte (HaCaT) cultures were exposed to 1 µg/ml LPS for 8 h, and then adalimumab was added to the cultures at a concentration of 8 µg/ml and incubated for 2, 8, and 24 h. We used mRNA and miRNA microarray, quantitative reverse transcription polymerase chain reaction, and enzyme-linked immunosorbent assay techniques. Results: STAT1, STAT3, STAT5, IL-6, IL-6R, SOCS3, and JAK3 genes differentiated HaCaT cultures with the drug from controls regardless of the time the cells were exposed to the drug. The addition of adalimumab to a culture previously exposed to LPS resulted in silencing of SOCS3 and IL-6 expression compared to the control, while for the other transcripts they were found to be overexpressed compared to the control culture. The assessment indicated the strongest connections between JAK3 and hsa-miR-373-5p (target score 96); SOCS3, STAT5, and hsa-miR-1827 (target score 96). Conclusions: Our study indicates that adalimumab has the strongest modulating effect on mRNA and miRNA expression of JAK/STAT and IL-6-dependent IL-12/23 pathways.

Dynamics of Microbiome Changes in the Endometrium and Uterine Cervix during Embryo Implantation: A Comparative Analysis.

BACKGROUND The microbiome is the collection of all micro-organisms and their genes, which naturally live in and on the body. The cervical and endometrial bacterial microbiome has previously been reported to affect fertility and influence the outcomes of assisted reproductive therapy (ART), including embryo transfer. This study aimed to evaluate the cervical and endometrial bacterial microbiome in 177 women treated for infertility before, during, and after embryo implantation, and the outcomes. MATERIAL AND METHODS Cervical and endometrial swabs were collected from 177 women diagnosed with infertility at 3 time points: (1) during the initial examination, (2) during implantation, (3) 10-14 days after implantation. Next-generation sequencing (NGS) was used to analyze the bacterial microbiome. Taxonomic identification was performed with the Usearch algorithm. RESULTS There was a significant change in the number of patients with Escherichia coli depending on the collection time. For the first swab collection, there were significant negative relationships between the percentage of Gardnerella vaginalis and Lactobacillus spp. For the second collection, there was a negative relationship between Lactobacillus helveticus and Lactobacillus jensenii. For the third collection, negative relationships were found between Escherichia coli and Lactobacillus spp. A similar distribution of the bacterial microbiome was observed in all 3 swab collections. CONCLUSIONS Lactobacillus spp. were the main bacteria identified in the cervix and endometrium, present before, during, and after successful embryo transfer. E. coli and G. vaginalis reduced the protective effect of Lactobacilli before, during, and after embryo transfer.

Detection and Genotyping of Human Papillomavirus (HPV16/18), Epstein-Barr Virus (EBV), and Human Cytomegalovirus (HCMV) in Endometrial Endometroid and Ovarian Cancers.

The purpose of this study was to evaluate the relationship between human papillomavirus (HPV16/18), Epstein-Barr virus (EBV), and human cytomegalovirus (HCMV) infections and the occurrence of ovarian cancer in 48 women, of whom 36 underwent surgery and chemotherapy (group A), 12 in whom surgery was sufficient (group B), and 60 with endometroid endometrial cancer stage G1-G3 (group C), compared to patients in whom the uterus and its appendages were removed for nononcological reasons (control group). The detection of HPV, EBV, and HCMV in tumor tissue and normal tissue was performed using the real-time polymerase chain reaction (RT-PCR) technique. A statistically significantly higher risk of endometrial cancer was noted in patients infected only with HCMV (OR > 1; p < 0.05). In contrast, a significantly higher risk of ovarian cancer in group A was associated with HPV16, HPV18, and EBV (OR > 1; p < 0.05); a significantly higher risk of ovarian cancer in group B was associated with HPV18 and HMCV (OR > 1; p < 0.05). The obtained results suggest that HCMV infection is associated with the development of a stage of ovarian cancer when treatment can be completed with surgery alone. Meanwhile, EBV appears to be responsible for the development of ovarian cancer in more advanced stages.

Reinfections from SARS-CoV-2: A Retrospective Study from the Gyncentrum Genetic Laboratory in Sosnowiec, Poland, April 2020 to July 2022.

BACKGROUND The increasing number of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) reinfections has opened a new research direction related to analyzing long-term immune response and accurately characterizing individual cases of reinfection to understand its mechanism and estimate the risk of widespread reinfection both locally and globally. This retrospective study from the Gyncentrum Genetic Laboratory in Sosnowiec, Poland aimed to evaluate reinfections from SARS-CoV-2 between April 2020 and July 2022. MATERIAL AND METHODS The study extended the previously published report on SARS-CoV-2 infection cases in Poland by analyzing 8041 reinfections diagnosed with real-time reverse transcription-polymerase chain reaction (RT-PCR) assay. Data were collected on the amount of time elapsed from the first infection to the next and, based on these data, all results were divided into several groups for statistical analysis: 0-44, 45-90, 91-200, 201-310, 311-420, and >420 (days). RESULTS The study showed that of the 8041 patients who experienced reinfection, the vast majority (5505) became reinfected more than 310 days after the original infection, even though the average time between infections was 354.3 days. Statistical analysis revealed that the risk of SARS-CoV-2 reinfection increases with time and that this relationship becomes statistically significant after the 200th day following the initial infection (p<0.01). CONCLUSIONS We have shown that acquired immunity to SARS-CoV-2 infection is relatively short-lived - it starts diminishing about 6 months after the initial positive test. Moreover, the risk of reinfection is very high more than 1 year after the initial infection.

miRNAs Participate in the Regulation of Oxidative Stress-Related Gene Expression in Endometrioid Endometrial Cancer.

Reactive oxygen species are formed as by-products of normal cell metabolism. They are needed to maintain cell homeostasis and signaling, which is possible due to defense systems. Disruption of this balance leads to oxidative stress that can induce cancer. Redox regulation by miRNAs may be a potential therapeutic target. The aim of the study was to assess the activity of genes associated with oxidative stress in endometrial cancer and to determine their relationship with miRNAs. The study included 45 patients with endometrioid endometrial cancer and 45 without neoplastic changes. The expression profile of genes associated with oxidative stress was determined with mRNA microarrays, RT-qPCR and ELISA. The miRNA prediction was performed based on the miRNA microarray experiment and the mirDB tool. PRDX2 and AQP1 showed overexpression that was probably not related to miRNA activity. A high level of PKD2 may be the result of a decrease in the activity of miR-195-3p, miR-20a, miR-134. A SOD3 level reduction can be caused by miR-328, miR-363. In addition, miR-363 can also regulate KLF2 expression. In the course of endometrial cancer, the phenomenon of oxidative stress is observed, the regulation of which may be influenced by miRNAs.

Nutritional Strategies for the Individualized Treatment of Non-Alcoholic Fatty Liver Disease (NAFLD) Based on the Nutrient-Induced Insulin Output Ratio (NIOR).

Nutrients play a fundamental role as regulators of the activity of enzymes involved in liver metabolism. In the general population, the action of nutrients may be affected by gene polymorphisms. Therefore, individualization of a diet for individuals with fatty liver seems to be a fundamental step in nutritional strategies. In this study, we tested the nutrient-induced insulin output ratio (NIOR), which is used to identify the correlation between the variants of genes and insulin resistance. We enrolled 171 patients, Caucasian men (n = 104) and women (n = 67), diagnosed with non-alcoholic fatty liver disease (NAFLD). From the pool of genes sensitive to nutrient content, we selected genes characterized by a strong response to the NIOR. The polymorphisms included Adrenergic receptor (b3AR), Tumor necrosis factor (TNFα), Apolipoprotein C (Apo C III). Uncoupling Protein type I (UCP-1), Peroxisome proliferator activated receptor γ2 (PPAR-2) and Apolipoprotein E (APOEs). We performed three dietary interventions: a diet consistent with the results of genotyping (NIOR (+)); typical dietary recommendations for NAFLD (Cust (+)), and a diet opposite to the genotyping results (NIOR (-) and Cust (-)). We administered the diet for six months. The most beneficial changes were observed among fat-sensitive patients who were treated with the NIOR (+) diet. These changes included improvements in body mass and insulin sensitivity and normalization of blood lipids. In people sensitive to fat, the NIOR seems to be a useful tool for determining specific strategies for the treatment of NAFLD.

Metabolic risk factors of coronary heart disease in relation to anthropometric measures in nonalcoholic fatty liver disease patients following dietary intervention.

Introduction: Nonalcoholic fatty liver disease (NAFLD) is connected with high risk of metabolic disorders, such as: metabolic syndrome, type 2 diabetes, dyslipidemia and independently, coronary heart disease (CHD). Materials and methods: A group of 108 Caucasian individuals (39 women and 69 men) aged 20­77 years with NAFLD were prospectively enrolled in the study. The diagnosis of NAFLD was con- ô€irmed during the ô€irst appointment using ultrasonography. The liver serum tests such as alanine aminotransferase and aspartate aminotransferase levels were taken into account, as well. All of the tests and measurements were taken at the begining and after 6 month of dieting. Anthropometric tools (body mass index ­ BMI, waist circumference ­ WC, waist to hip ratio ­ WHR, waist to height ratio ­ WHtR, coinicity index ­ C -Index) were examined in relation to metabolic risk factors of CHD (increased: triacylglycerols, low density lipoprotein, total cholesterol, glycemia and low high density lipoprotein). Results: Statistical signiô€icant reduction of body weight, BMI, WC, WHR, WHtR and C -Index (p < 0.05) contributed to the improvement of serum triacylglycerols in both sexes (p < 0.05). Indicators of abdominal obesity (WC, WHR, WHtR, C -Index) correlated signiô€icantly with serum glucose (p < 0.05) both before and after the diet and with serum lipids, either before or after the diet (p < 0.05), in both sexes. Conclusions: It seems beneô€icial to spread the use of anthropometric tools, especially C -Index and WHtR, in primary health care practice for either early prevention or prediction of metabolic CHD risk occurrence, and its potential fatal consequences among patients with NAFLD or with other metabolic disorders. Keywords: fatty liver disease; cardiovascular disease; risk factors; anthropometry; nutrition; metabolic syndrome.

Metabolites of arachidonic acid and linoleic acid in early stages of non-alcoholic fatty liver disease--A pilot study.

BACKGROUND: Nonalcoholic fatty liver disease (NAFLD) is a spectrum of liver conditions related to fat infiltration. The role of liver triacylglycerol accumulation in NAFLD is not fully understood. METHODS: Twenty-four patients, 12 in the first and 12 in the second stage of NAFLD, were prospectively enrolled in this study. Biochemical parameters and eicosanoids (HETE and HODE) were compared between the first and the second stage of hepatic steatosis and the effect of a 6-month dietary intervention on these parameters was evaluated. Eicosanoid profiles were extracted from 0.5 ml of plasma using solid-phase extraction RP-18 SPE columns. The HPLC separations were performed on a 1260 liquid chromatograph. RESULTS: Patients with stage I NAFLD had a significantly higher level of HDL cholesterol and a lower level of 5-HETE. Patients with grade II steatosis had higher concentrations of 9-HODE. Following the six-month dietary intervention, hepatic steatosis resolved completely in all patients. This resulted in a significant decrease in the concentrations of all eicosanoids (LX4, 16-HETE, 13-HODE, 9-HODE, 15-HETE, 12-HETE, 5-oxoETE, 5-HETE) and key biochemical parameters (BMI, insulin, HOMA-IR, liver enzymes). CONCLUSION: A significant reduction in the analyzed eicosanoids and a parallel reduction in fatty liver confirmed the usefulness of HETE and HODE in the assessment of NAFLD.

Fatty acid changes help to better understand regression of nonalcoholic fatty liver disease.

AIM: To investigate whether liver steatosis reduction due to a six-month dietary intervention results in significant changes in the concentrations of fatty acids. METHODS: A group of 35 Caucasian individuals diagnosed with different levels of steatosis were prospectively enrolled in the present study. Analysis of the fatty acid profiles was performed according to changes in liver steatosis (liver steatosis reduction by one or two degrees) after a six-month dietary intervention. The diet helped reduce body mass in obese and overweight patients, and stabilize both glycemia and dyslipidemia. Fatty acids were extracted according to the Folch method and analyzed by gas chromatography. RESULTS: This study showed significant changes in fatty acid profiles in patients who had reduced liver steatosis by one as well as two degrees. A reduction in liver steatosis by one degree caused a significant increase in the level of the n-3 family: eicosapentaenoic acid (P < 0.055), docosapentaenoic acid-C 22:5 (P < 0.05) and docosahexaenoic acid (P < 0.05). A reduction in liver steatosis by two degrees caused a significant decrease in serum palmitoleic acid-C 16:1 (P < 0.05). CONCLUSION: Liver steatosis reduction is associated with changes in fatty acid profiles, and these changes may reflect an alteration in fatty acid synthesis and metabolism. These findings may help better understand regression of nonalcoholic fatty liver disease.

[Carbon monoxide poisoning in pregnant woman]. / Zatrucie tlenkiem wegla u kobiety ciezarnej.

Carbon monoxide poisoning is one of the most frequent types of poisoning caused by gases. Exposure of a pregnant woman to carbon monoxide is connected with transmission of this gas to her fetus what may cause oxygen deficiency, and further, the damage to its organs. The article describes a case of carbon monoxide poisoning of a 28-weeks pregnant woman who was treated in a hyperbaric chamber. Therapy was successful and the woman gave birth to a healthy, full term infant. In case of poisoning to a pregnant woman the above seems to be the most advantageous solution.

Usefulness of marking alkaline phosphatase and C-reactive protein in monitoring the risk of preterm delivery.

BACKGROUND: The purpose of this paper is to compare the effectiveness of use of alkaline phosphatase (ALP) and C-reactive protein (CRP) levels in marking and monitoring the risk of preterm delivery due to infection. MATERIAL/METHODS: The study involved 83 patients assigned to groups: Group I (n=43) consisted of patients hospitalized for symptoms of preterm delivery, and Group II (control group n=40) consisted of patients controlled or hospitalized delivering on time without complications, whose pregnancy had a physiological course. All patients had a single marking of ALP and CRP levels in serum performed. RESULTS: CRP levels were within the range 7 mg/l to 94 mg/l in the study group, and 4.83 mg/l to 90 mg/l in the control group. The level of ALP in the study group ranged from 139 u/l to 368 u/l and from 218 u/l to 321 u/l in the control group. In more than half of women (72.1%) from study group, CRP level exceeded 7 mg/l; in the control group, the CRP level exceeded 7 mg/l in 35% of cases. Significantly higher levels of CRP (above 20 mg/l) and ALP (above 300 u/l) were found in the 18 patients from the study group compared to the control group. CONCLUSIONS: Although an increase in the level of ALP in serum cannot be an absolute and sole marker of the risk of preterm delivery, it can be used in conjunction with a significantly elevated CRP level.

Doppler velocimetry of the materno-fetal circulation in preterm delivered pregnancies complicated with hypertension.

BACKGROUND: Hypertension is one of the most frequent complications of pregnancy. Due to high risk of morbidity and mortality in both mothers and children, it is necessary to continuously monitor the pregnancy, principally with biophysical methods. Particularly, doppler velocimetry of the materno-fetal circulation proves useful. THE AIM of the study was to assess the usefulness of doppler test in monitoring the condition of the foetus in preterm delivered pregnancy complicated with hypertension. MATERIAL AND METHODS: The retrospective analysis comprised the data of 116 women who delivered prematurely at the Clinics of the Department of Gynaecology and Obstetrics at the Collegium Medicum of the Jagiellonian University in the years 2006-2007, resulting in creation of Group I involving 38 pregnant women with preeclampsia, and Group II of 36 women whose pregnancy was complicated with gestational hypertension. Control group was formed of 42 women with correct arterial blood pressure. When describing the groups, the differences in the birth weight and Apgar score were indicated. RESULTS: A significant statistical difference was found in the area of pulsation rate in the umbilical artery and cerebro-placental ratio (CPR). In the case of preterm delivery complicated with arterial pressure disorders, the foetus is characterised with worse organic perfusion and slower somatic growth than if no concomitant hypertension is present. Hypertension forms an additional risk factor in the course of preterm delivery, and doppler velocimetry is a good method for monitoring the condition of the foetus, as it allows for detection of irregularities and for implementation of relevant treatment to improve the newborn's condition at birth.

[Surgical strategy and outcome in multiple cerebral aneurysms]. / Taktyka postepowania chirurgicznego i wyniki leczenia tetniaków mnogich mózgu.

INTRODUCTION: Multiple aneurysms of the brain are a more dangerous disorder than single aneurysms. Most of patients have evidence of aneurysmal rupture, but rebleeding remains an additional danger because of co-existing aneurysms. These "silent" aneurysms also have to be operated--the risk of following subarachnoid haemorrhage (SAH) is about 2% annually. AIM: The purpose of this study was to assess of the surgical outcome in multiple aneurysmal SAH in regard to chosen risk factors. MATERIAL AND METHOD: Retrospective analysis of 608 patients treated within three years (2001-2003) for cerebral aneurysms in the Department of Neurosurgery, Jagiellonian University in Kraków, was carried out. 14% patients had multiple aneurysms: 72--two aneurysms, 10 patients three aneurysms, and three patients four and more. The mean age was 50, with slight predominance of women (58%). Most patients were operated on via pterional craniotomy, and posterior circulation aneurysms were clipped by means of suboccipital approach. "Silent" aneurysms being clipped in the first stage, if were available via the same craniotomy, another--in the next stage. We analyzed short-term outcome of multiple aneurysms in comparison to the single (literature data). We presented the outcome (according to the Glasgow Outcome Scale) with regard to preoperative condition, age, location of the most frequent "bleeding" aneurysms (middle cerebral artery, anterior communicating artery), and in patients operated on in the 1st and 2nd stage. Accuracy of the preoperative detection of "bleeding" aneurysm was checked based on CT, MRI, angio-CT, angio-MRI and DSA. RESULTS AND CONCLUSIONS: Out of all 85 treated patients, 64 (75.3%) presented very good and good condition on discharge. Five (5.9%) were severe disabled, and 16 (18.8%) died. Good preoperative condition, age less than 55, and "bleeding" aneurysms of the middle cerebral artery (compared with anterior communicating artery) are good prognostic factors in surgically treated multiple aneurysms. Outcome in single and multiple aneurysms is similar, excluding perioperative losses, which are 5% higher in the multiple aneurysms group. In our series in each case of "bleeding" aneurysm was correctly detected preoperatively.