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.

The influence of selected microRNAs on the expression profile of genes and proteins related to the tumor necrosis factor-alpha signaling pathways in endometrioid endometrial cancer.

PURPOSE: Tumor necrosis factor exerts many adverse biological effects, from cell proliferation to cell death. Accurate diagnosis and treatment are therefore difficult due to many factors influencing tumor necrosis factor-alpha (TNF-α) signaling, including microRNAs (miRNAs), especially in tumors. The aim of the study was to determine the influence of miRNAs on the expression profile of genes and proteins related to TNF-α signaling in endometrial cancer. METHODS: The material consisted of 45 endometrioid endometrial cancer and 45 normal endometrium tissue samples. Gene expression was determined with microarrays and then validated for TNF-α, tumor necrosis factor receptor 1 (TNFR1) and 2 (TNFR2), caveolin 1 (CAV1), nuclear factor kappa B subunit 1 (NFKB1), and TGF-beta activated kinase 1 (MAP3K7)-binding protein 2 (TAB2) using real-time quantitative reverse transcription reaction (RT-qPCR). The protein concentration was assessed by enzyme-linked immunosorbent assay (ELISA). In addition, differentiating miRNAs were identified using miRNA microarrays and their relationships with TNF-α signaling genes were evaluated using the mirDIP tool. RESULTS: TNF-α, TNFR1, TNFR2, CAV1, NFKB1, and TAB2 were upregulated both on the mRNA and protein levels. The decrease in the activity of miR-1207-5p, miR-1910-3p, and miR-940 may be related to CAV1 overexpression. Similarly for miR-572 and NFKB1 as well as miR-939-5p and TNF-α. In turn, miR-3178 may partially inhibit TNFR1 activity up to grade 2 cancer. CONCLUSION: TNF-α signaling, especially the TNF-α/NF-κB axis, is disrupted in endometrial cancer and worsens with disease progression. The observed changes may be the result of miRNAs' activity in the initial stage of endometrial cancer and its gradual loss in later grades.

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.

Daily levels of sex hormones in 15 subfertile women formulate a menstrual cycle profile predominant with progesterone secretion.

INTRODUCTION: Changes in sex hormone secretions during the menstrual cycle may affect fertility. It has been shown that a prematurely raised progesterone (P4) level after therapeutic injection of human chorionic gonadotropin caused changes in endometrial gene expression and lowered the pregnancy rate. The aim of the present study was to investigate the complete menstrual patterns of P4 together with its derivatives testosterone (T) and oestradiol (E2) in subfertile women during their natural cycles. MATERIAL AND METHODS: Daily serum levels of P4 (ng/mL), T (ng/mL), E2 (pg/mL), and sex hormone binding protein (SHBG, nmol/L) were measured throughout a single 23-28-day menstrual cycle in 15 subfertile women aged 28-40 years with patent oviducts and normospermic partners. Knowing SHBG levels, the free androgen (FAI) and free oestrogen (FEI) indexes were calculated for each cycle day in each patient. RESULTS: Baseline (cycle day one) levels of luteinising hormone (LH), thyroid stimulating hormone (TSH), P4, and T were comparable with reference intervals for a normal cycle, whereas follicle stimulating hormone (FSH), E2, and SHBG exceeded those. During cycles, the levels of P4 correlated positively with E2 levels (r = 0.38, p < 0.05, n = 392) an negatively with T (r = -0.13, p < 0.05, n = 391). T correlated negatively with E2 (r = -0.19, p < 0.05, n = 391). Menstrual cycle phases were hidden. The curve of the mean/median daily levels of P4 rose prematurely, was parallel with the E2 rise, and culminated closely, but with more than 4 times greater amplitude of P4 (2571% of baseline levels in day 16) than of E2 (580% in day 14). In turn, the curve of T declined in a U-shaped manner with a nadir (-27%) on day 16. Averaged daily levels of FEI, but not FAI, varied significantly between 23 and 26 days long and the 27-28-day cycles. CONCLUSIONS: 1. Throughout the entire menstrual cycle length in subfertile women, P4 secretion predominates quantitatively over secretions of the remaining sex hormones when menstrual cycle phases are hidden. 2. The rise of E2 secretion is in parallel with the P4 rise, but with 4 times lower amplitude of E2. 3. T secretion declines and is inversely related to both P4 and E2 secretions. 4. Changes in E2 bioavailability are related to menstrual cycle length.

A Retrospective Population Study of 385 191 Positive Real-Time Reverse Transcription-Polymerase Chain Reaction Tests For SARS-CoV-2 from a Single Laboratory in Katowice, Poland from April 2020 to July 2022.

BACKGROUND This retrospective population study identified 385 191 positive real-time reverse transcription-polymerase chain reaction (RT-PCR) tests for the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) from a single laboratory in Katowice, Poland, from April 2020 to July 2022. MATERIAL AND METHODS The material was nasopharyngeal, nasopharyngeal swab or bronchial lavage, and bronchoalveolar lavage (BAL) to confirm or exclude SARS-CoV-2 infection with the RT-PCR technique. Personal data are use according to the Provisions on the Protection of Personal Data by the Gyn-Centrum laboratory. RESULTS In 9 months of 2020, the number of SARS-CoV-2 results was 88 986; in 2021, it was 168 439, and in the first 7 months of 2022, it was 12 786. In 2020, the highest number of positive results was recorded in the third quarter (83 094 cases); 2021, in the 1st, 2nd, and 4th quarters (58 712; 37 720; and 71 753 cases, respectively), and in 2022, in the 1st quarter (127 613 cases) of the year. A positive result was observed more often in women and people aged 30-39, followed by those 40-49 years. Patients aged 10-19 years comprised the smallest population of SARS-CoV-2-positive cases. CONCLUSIONS In the Polish population studied, from April 2020 to July 2022, the detection rates of SARS-CoV-2 positivity were significantly higher for women than for men and in the 30-49 age group for both sexes. Also, the infection detection rate of 385 191 out of 1 332 659 patient samples, or 28.9%, supports that the Polish society adhered to public health recommendations for infection control during the COVID-19 pandemic.

Clustered PHD domains in KMT2/MLL proteins are attracted by H3K4me3 and H3 acetylation-rich active promoters and enhancers.

Histone lysine-specific methyltransferase 2 (KMT2A-D) proteins, alternatively called mixed lineage leukemia (MLL1-4) proteins, mediate positive transcriptional memory. Acting as the catalytic subunits of human COMPASS-like complexes, KMT2A-D methylate H3K4 at promoters and enhancers. KMT2A-D contain understudied highly conserved triplets and a quartet of plant homeodomains (PHDs). Here, we show that all clustered (multiple) PHDs localize to the well-defined loci of H3K4me3 and H3 acetylation-rich active promoters and enhancers. Surprisingly, we observe little difference in binding pattern between PHDs from promoter-specific KMT2A-B and enhancer-specific KMT2C-D. Fusion of the KMT2A CXXC domain to the PHDs drastically enhances their preference for promoters over enhancers. Hence, the presence of CXXC domains in KMT2A-B, but not KMT2C-D, may explain the promoter/enhancer preferences of the full-length proteins. Importantly, targets of PHDs overlap with KMT2A targets and are enriched in genes involved in the cancer pathways. We also observe that PHDs of KMT2A-D are mutated in cancer, especially within conserved folding motifs (Cys4HisCys2Cys/His). The mutations cause a domain loss-of-function. Taken together, our data suggest that PHDs of KMT2A-D guide the full-length proteins to active promoters and enhancers, and thus play a role in positive transcriptional memory.

Lung Cancer Risk in Men and Compliance with the 2018 WCRF/AICR Cancer Prevention Recommendations.

Lung cancer is the most common and deadly form of cancer worldwide, especially in men. The 2018 World Cancer Research Fund/American Institute for Cancer Research (WCRF/AICR) updated cancer prevention recommendations, and a standard scoring system (2018 WCRF/AICR Score) was published. The purpose of this study was to develop the adapted version of the 2018 WCRF/AICR Score with respect to lung cancer prevention recommendation (Ad-LC WCRF/AICR Score) and to examine the association between lung cancer risk in men and the Ad-LC WCRF/AICR Score as well as its single components. A case-control study was conducted among 439 men aged 45-80 years (187 controls, 252 primary lung cancer cases). Lifestyle and dietary data were collected with a questionnaire including the 62-item food frequency questionnaire (FFQ-6®). The Ad-LC WCRF/AICR Score was used as a categorized and continuous variable. Odds ratios (ORs) and 95% confidence intervals (95% CIs) for lung cancer risk were calculated with the partly and fully adjusted model. One component of the score was independently associated with a lower risk of lung cancer in men, regardless of the set of confounders used. In the fully adjusted model following the recommendation "Limit smoking" was associated with a lower risk of lung cancer-in the never smokers by 87% (OR: 0.13; 95% CI: 0.04-0.37; p = 0.0002) and in the moderate smokers by 45% (OR: 0.55; 95% CI: 0.33-0.91; p = 0.0189) compared with the heavy smokers as a reference. By adding the single components making up the Ad-LC WCRF/AICR Score, the combination of three components or more, reducing the risk of lung cancer compared to lower compliance as a reference by 45% to 78% and by 39% to 66% for intermediate compliance (except two models out of seven) and higher compliance, respectively. In the fully adjusted model, the risk of lung cancer for the total Ad-LC WCRF/AICR Score was lower by 47% (OR: 0.53; 95% CI: 0.32-0.88; p = 0.0129) in higher compliance with the score compared to those with the lower compliance. Each one-point increase in the Ad-LC WCRF/AICR Score reduced lung cancer risk by 34% (OR: 0.66; 95% CI: 0.45-0.95; p = 0.0267). The results support previous evidence that limiting smoking reduces the risk of lung cancer in men. It also provides an insight into cancer research by showing that following the combined 2018 cancer prevention recommendations related to diet, lifestyle and body fatness was associated with a lower risk of lung cancer in men.

Semen dysbiosis-just a male problem?

Seminal microflora is crucial to male fertility. Dysbiosis-disturbance of quantitative ratios of individual bacteria or appearance of pathogenic species-rarely results in symptomatic disease. Inflammation results in decreased sperm production, lower motility, or morphological changes and, in the long term, can cause ejaculatory duct obstruction, leading to infertility. Moreover, it may cause infection of the partner's female genital tract. Dysbiosis in both partners results in fertility problems, disorders in embryo implantation, or miscarriages. In addition, chronic inflammation of the male genitourinary system may accelerate the appearance of antisperm antibodies. A comprehensive examination of seminal microflora can clarify the causes of infertility or prevent pathological conditions that affect seminal parameters. Seminal microflora as a direct impact on fertility problems as well as a decrease in the effectiveness of assisted reproduction methods, insemination, or in vitro procedures.

From Alpha to Delta-Genetic Epidemiology of SARS-CoV-2 (hCoV-19) in Southern Poland.

In Poland, the first case of SARS-CoV-2 infection was confirmed in March 2020. Since then, many circulating virus lineages fueled rapid pandemic waves which inflicted a severe burden on the Polish healthcare system. Some of these lineages were associated with increased transmissibility and immune escape. Mutations in the viral spike protein, which is responsible for host cell recognition and serves as the primary target for neutralizing antibodies, are of particular importance. We investigated the molecular epidemiology of the SARS-CoV-2 clades circulating in Southern Poland from February 2021 to August 2021. The 921 whole-genome sequences were used for variant identification, spike mutation, and phylogenetic analyses. The Pango B.1.1.7 was the dominant variant (n = 730, 89.68%) from March 2021 to July 2021. In July 2021, the B.1.1.7 was displaced by the B.1.617.2 lineage with 66.66% in July 2021 and 92.3% in August 2021 frequencies, respectively. Moreover, our results were compared with the sequencing available on the GISAID platform for other regions of Poland, the Czech Republic, and Slovakia. The analysis showed that the dominant variant in the analyzed period was B.1.1.7 in all countries and Southern Poland (Silesia). Interestingly, B.1.1.7 was replaced by B.1.617.2 earlier in Southern Poland than in the rest of the country. Moreover, in the Czech Republic and Slovakia, AY lineages were predominant at that time, contrary to the Silesia region.

Marking the Profile of the Microflora of the Endometrium and Uterine Cervix in Women as a Potential Factor Determining the Effectiveness of In Vitro Fertilization.

One promising research trend involves evaluating the influence of microbiota in the reproductive system of women on becoming pregnant and maintaining pregnancy. The goal of this study was to define the microflora profile of the endometrium and uterine cervix in women qualified for an in vitro fertilization (IVF) procedure, which is expected to contribute to increasing the percentage of successful IVF implantations. Based on the conducted molecular analysis in the collected swabs, 22 bacterial strains were identified. Eleven strains (57%) that were isolated belong to the physiological microflora, the most common strain of which was Lactobacillus. Eight of the isolated strains (33%) were pathological microflora, among which the most common bacteria were from the Enterobacteriaceae family (which includes E. coli, Shigella, and Salmonella). Finally, three of the bacterial strains (10%) may be a component of both physiological or pathological microflora of the vagina: Bifidobacterium breve, Bifidobacterium longum group, and Alloscardovia omnicolens. The presence of Escherichia coli was detected in six women, Staphylococcus aureus also in six patients, Atopobium parvulum in three, Streptococcus salivarius group in three, Enterococcus faecalis in four, and Aerococcus christensenii in two patients. We found statistically significant relationships (p < 0.05) between Lactobacillus fermentum and Enterococcus faecalis, Lactobacillus delbrueckii and Escherichia coli groups, Lactobacillus FN667084_s and Staphylococcus aureus groups, as well as Lactobacillus fermentum and Streptococcus agalactiae. Based on the conducted study, it may be confirmed that the endometrium is, to a large extent, colonized by lactic acid bacilli. Apart from that, endometrial dysbiosis was not noted in patients qualified for the IVF procedure.

Signaling mechanisms and their regulation during in vivo or in vitro maturation of mammalian oocytes.

In vitro fertilization (IVF) is currently one of the most effective methods of infertility treatment. An alternative to commonly used ovarian hyperstimulation can become extracorporeal maturation of oocytes (in vitro maturation; IVM). Fertilization and normal development of the embryo depends on the cytoplasmic, nuclear and genomic maturity of the oocyte. The microenvironment of the ovarian follicle and maternal signals, which mediate bidirectional communication between granulosa, cumulus and oocyte cells, influence the growth, maturation and acquisition of oocyte development capability. During oogenesis in mammals, the meiosis is inhibited in the oocyte at the prophase I of the meiotic division due to the high cAMP level. This level is maintained by the activity of C-type natriuretic peptide (CNP, NPPC) produced by granulosa cells. The CNP binds to the NPR2 receptor in cumulus cells and is responsible for the production of cyclic guanosine monophosphate (cGMP). The cGMP penetrating into the oocyte through gap junctions inhibits phosphodiesterase 3A (PDE3A), preventing cAMP hydrolysis responsible for low MPF activity. The LH surge during the reproductive cycle reduces the activity of the CNP/NPR2 complex, which results in a decrease in cGMP levels in cumulus cells and consequently in the oocyte. Reduced cGMP concentration unblocks the hydrolytic activity of PDE3A, which decreases cAMP level inside the oocyte. This leads to the activation of MPF and resumption of meiosis. The latest IVM methods called SPOM, NFSOM or CAPA IVM consist of two steps: prematuration and maturation itself. Taking into account the role of cAMP in inhibiting and then unblocking the maturation of oocytes, they have led to a significant progress in terms of the percentage of mature oocytes in vitro and the proportion of properly developed embryos in both animals and humans.

Endocrine disorders and fertility and pregnancy: An update.

It is estimated that more and more couples suffer from fertility and pregnancy maintenance disorders. It is associated with impaired androgen secretion, which is influenced by many factors, ranging from genetic to environmental. It is also important to remember that fertility disorders can also result from abnormal anatomy of the reproductive male and female organ (congenital uterine anomalies - septate, unicornuate, bicornuate uterus; acquired defects of the uterus structure - fibroids, polyps, hypertrophy), disturbed hormonal cycle and obstruction of the fallopian tubes resulting from the presence of adhesions due to inflammation, endometriosis, and surgery, abnormal rhythm of menstrual bleeding, the abnormal concentration of hormones. There are many relationships between the endocrine organs, leading to a chain reaction when one of them fails to function properly. Conditions in which the immune system is involved, including infections and autoimmune diseases, also affect fertility. The form of treatment depends on infertility duration and the patient's age. It includes ovulation stimulation with clomiphene citrate or gonadotropins, metformin use, and weight loss interventions. Since so many different factors affect fertility, it is important to correctly diagnose what is causing the problem and to modify the treatment regimen if necessary. This review describes disturbances in the hormone secretion of individual endocrine organs in the context of fertility and the maintenance of pregnancy.

Evaluation of the Differences in the Expression of Biogenic Amine-Related mRNAs and Proteins in Endometrioid Endometrial Cancer.

Biogenic amines, such as adrenaline, noradrenaline, histamine, dopamine, and serotonin are important neurotransmitters that also regulate cell viability. Their detection and analysis are helpful in the diagnosis of many diseases, including cancer. The aim of this study was to determine the expression profile of the biogenic amine-related genes and proteins in endometrioid endometrial cancer compared to the control group. The material consisted of endometrial tissue samples and whole blood collected from 30 endometrioid endometrial cancer patients and 30 cancer-free patients. The gene expression was determined by the mRNA microarrays and validated by qRT-PCR. Protein levels were determined in the serum by the enzyme-linked immunosorbent assay (ELISA). Overexpression of histamine H1-H3 receptors and early growth response 1 and silencing of calmodulin, the histamine H4 receptor, and the dopamine D5 receptor have been reported in endometrioid endometrial cancer. The obtained results indicate disturbances in the signaling activated by histamine and dopamine receptors, which could potentially contribute to the progression of endometrioid endometrial cancer.

miRNAs in the Expression Regulation of Dopamine-Related Genes and Proteins in Endometrial Cancer.

Disruption of the dopaminergic system leads to many diseases, including cancer. Dopamine and its receptors are involved in the regulation of proliferation, cell death, invasion, and migration. Better understanding of the mechanisms involved in these processes could reveal new molecular markers and therapeutic targets. The aim of this study was to determine the expression profile of dopamine-related genes and proteins in endometrial cancer and to assess whether miRNAs are involved in its regulation. Sixty women were recruited for the study: 30 with endometrial cancer and 30 without cancer. The expression profiles of dopamine-related genes were determined in endometrial tissue samples using microarrays and qRT-PCR. Then, protein concentration was determined with the ELISA test. In the last step, miRNA detection was performed using microarrays. The matching of miRNAs to the studied genes was carried out using the TargetScan tool. The analysis showed DRD2 and DRD3 overexpression, with a reduction in DRD5 expression, which could be due to miR-15a-5p, miR-141-3p, miR-4640-5p, and miR-221-5p activity. High levels of OPRK1 and CXCL12, related to the activity of miR-124-3p.1 and miR-135b-5p, have also been reported. Low COMT expression was probably not associated with miRNA regulation in endometrial cancer.

Variances in the Expression of mRNAs and miRNAs Related to the Histaminergic System in Endometrioid Endometrial Cancer.

Research has indicated higher concentrations of histamine and polyamine in endometrioid tissue in comparison with healthy tissue. The aim of this study was to evaluate changes in the expression patterns of messenger RNA (mRNAs) and microRNA (miRNAs) related to the histaminergic system in endometrial samples and whole blood in women with endometrioid endometrial cancer. The study group consisted of 30 women with endometrioid endometrial cancer qualified for hysterectomy (G1 well-differentiated, 15 cases; G2 moderately differentiated, 8 cases; and G3 poorly differentiated, 7 cases). The control group included 30 women with no neoplastic changes during routine gynecological examinations. The molecular analysis consisted of the microarray analysis of mRNAs and miRNAs related to the histaminergic system, reverse-transcription quantitative polymerase chain reaction (RTqPCR), and enzyme-linked immunosorbent assay (ELISA). Out of 65 mRNAs connected with the histaminergic system, 10 differentiate the samples of tissue and blood obtained from patients with endometrioid endometrial cancer in comparison with the control group (p < 0.05). mRNA histamine receptor 1,3 (HRH1, HRH3), and solute carrier family 22 member 3 (SLC23A2) differentiating samples of endometrioid endometrial cancer independent of either G or control. The highest probability of interaction, based on the target score miRDB, between the selected miRNAs and mRNAs was found for the hybrids hsa-miR-1-3p and endothelin 1 (END1), hsa-miR-27a-5ß and SLC23A2. The selected mRNA and miRNA transcripts seem to be promising for molecularly targeted therapies in the context of endometrioid endometrial cancer.

Plasma gonadotropin levels in metformin-treated men with prediabetes: a non-randomized, uncontrolled pilot study.

Metformin was found to reduce elevated, but not normal, thyrotropin and prolactin levels. This non-randomized, uncontrolled pilot study investigated hypothalamic-pituitary-testicular axis activity in men with primary hypogonadism receiving metformin. The study population included 29 men with prediabetes, 10 of whom had been diagnosed with primary hypogonadism. Throughout the study, the participants were treated with metformin (2.55-3 g daily). Glucose homeostasis markers (fasting glucose, glycated hemoglobin, and HOMA1-IR), as well as circulating levels of follicle-stimulating hormone (FSH), luteinizing hormone (LH), testosterone, thyrotropin, prolactin, estradiol, and creatinine, were assessed at the beginning of the study and 16 weeks later. Both groups differed in baseline gonadotropin and testosterone levels. Fasting glucose, glycated hemoglobin, and HOMA1-IR were lower after than before metformin treatment. The changes in fasting glucose and HOMA1-IR were more pronounced in hypogonadal men than in subjects with testosterone levels within the reference range. Only in hypogonadal men, plasma concentrations of FSH and LH were lower at the end than at the beginning of the study. Levels of the remaining hormones remained unchanged throughout the study period. The reduction in FSH and LH levels correlated with their baseline levels and with the changes in HOMA1-IR. The results of our study suggest that metformin may decrease FSH and LH levels in men with hypergonadotropic hypogonadism.

Artificial miRNAs targeting CAG repeat expansion in ORFs cause rapid deadenylation and translation inhibition of mutant transcripts.

Polyglutamine (polyQ) diseases are incurable neurological disorders caused by CAG repeat expansion in the open reading frames (ORFs) of specific genes. This type of mutation in the HTT gene is responsible for Huntington's disease (HD). CAG repeat-targeting artificial miRNAs (art-miRNAs) were shown as attractive therapeutic approach for polyQ disorders as they caused allele-selective decrease in the level of mutant proteins. Here, using polyQ disease models, we aimed to demonstrate how miRNA-based gene expression regulation is dependent on target sequence features. We show that the silencing efficiency and selectivity of art-miRNAs is influenced by the localization of the CAG repeat tract within transcript and the specific sequence context. Furthermore, we aimed to reveal the events leading to downregulation of mutant polyQ proteins and found very rapid activation of translational repression and HTT transcript deadenylation. Slicer-activity of AGO2 was dispensable in this process, as determined in AGO2 knockout cells generated with CRISPR-Cas9 technology. We also showed highly allele-selective downregulation of huntingtin in human HD neural progenitors (NPs). Taken together, art-miRNA activity may serve as a model of the cooperative activity and targeting of ORF regions by endogenous miRNAs.

Adherence to Prudent and Mediterranean Dietary Patterns Is Inversely Associated with Lung Cancer in Moderate But Not Heavy Male Polish Smokers: A Case-Control Study.

Lung cancer is the most commonly diagnosed cancer in men worldwide. Studies regarding dietary patterns (DPs) and lung cancer are limited, with results remaining inconclusive, and the association of DPs with lung cancer in smokers is unclear. This study analyzed the associations between DPs, including the Polish-adapted Mediterranean diet (Polish-aMED) score, and lung cancer risk in Polish adult male smokers. This case-control study involved 439 men aged 45-80 years from northeastern Poland, including 187 newly diagnosed lung cancer cases. Dietary data was collected with a 62-item food frequency questionnaire (FFQ-6). Two approaches were applied to identify dietary patterns. The Polish-aMED score was calculated (hypothesis-driven approach) and a principal component analysis (PCA) was used to identify PCA-driven DPs (data-driven approach). A logistic regression analysis was performed to estimate the odds ratio (OR) and 95% confidence interval (95% CI) of the lung cancer risk associated with the adherence to DPs overall as well as for moderate (2.5-11 pack-years) and heavy (>11 pack-years) smokers. Among moderate smokers, the risk of lung cancer was lower by 41% (OR: 0.59; 95% CI: 0.39-0.90; p < 0.05; adjusted model) in the higher adherence to the prudent DP when compared to the lower adherence, and by 66% (OR: 0.34; 95% CI: 0.15-0.76; p < 0.05; adjusted model) in the high adherence (7-9 points) to the Polish-aMED score when compared to the low adherence (0-3 points). No significant association between the westernized traditional DP or the sweet dairy DP and lung cancer was revealed. In conclusion, the current study suggests that pro-healthy dietary patterns, including the Mediterranean pattern, may favour lower risk of lung cancer in moderate smokers, although it was not confirmed in heavy smokers.

From "Every Day" Hormonal to Oxidative Stress Biomarkers in Blood and Follicular Fluid, to Embryo Quality and Pregnancy Success?

BACKGROUND: Oxidative stress appears to be involved in oocyte growth and maturation that when impaired results in poor embryo quality and lower potential to implant. The biochemical microenvironment of the oocyte (follicular fluid (FF)) consists of hormones and other various substances regulating the balance between oxidants and antioxidants. AIM: The aim of this study was to examine the possible impact of selected biomarkers ("every day," hormonal biomarkers, enzymatic and nonenzymatic antioxidants, and also oxidative stress markers) in serum and FF, on embryo quality and pregnancy success in infertile women undergoing infertility treatment. METHODS: All 53 patients, mean age 34.7 ± 4.1 years, with serum AMH level ≥ 0.7 ng/mL, were diagnosed with idiopathic infertility. They were stimulated in short antagonist protocol, followed by in vitro fertilization (IVF-ICSI intracytoplasmatic sperm injection) and a single embryo transfer. Follicular fluid was aspirated from the first mature follicle. In statistical analyses the R software was used, then all data was assessed with the Shapiro-Wilk test, logistic regression, and later the receiver operating characteristic (ROC) curve was applied using "pROC" R package. RESULTS: We did not observe any correlation between AMH and embryo quality and pregnancy rate. Statistically significant results were only found for biomarkers examined in follicular fluid. Greater levels of GPX in FF were associated with the increased chance of producing a high quality embryo (the optimal cut-off concentration was established at over 450 lU/L.) Regarding pregnancy success, increasing levels of GR (cut-off at 21 IU/L), CuZnSOD (cut-off at 9NU/mL), and GST (cut-off at 2.5 IU/L) resulted in lower chances of a successful pregnancy. CONCLUSION: Our results indicate that FF markers may have some advantages in predicting embryo quality and pregnancy over AMH. The GPX system seems to be mostly related to embryo quality and pregnancy.

Vitamin D and Anti-Müllerian Hormone Levels in Infertility Treatment: The Change-Point Problem.

BACKGROUND: Anti-Müllerian hormone (AMH) is considered to be one of the most significant indicators of women's fertility. Many studies have shown that vitamin D may modify human reproductive functions; however, the results are conflicting. The composition of follicular fluid (FF) creates the biochemical environment of the oocyte and affects its quality, which later determines the embryo quality. In this study, we aimed to revise with advanced statistical techniques the relationship between AMH and vitamin D in FF. METHODS: The study was designed as a prospective single-center study in infertile patients with AMH ≥ 0.7 ng/mL who underwent controlled ovarian hyperstimulation for in vitro fertilization. AMH and vitamin D levels in FF were measured. Next, the standard and advanced statistical (including segmented regression) techniques were applied. RESULTS: We observed a negative linear correlation between levels of AMH in serum and FF and total vitamin D concentrations up to approximately 30 ng/ml; with a statistically significant relationship in FF. Beyond that concentration, the trend was positive but statistically insignificant. CONCLUSIONS: As an existing "change-point problem" was noticed, we suggest segmentation in the relationship between vitamin D and AMH during infertility treatment.