Estrogen α and ß Receptor Expression in the Various Regions of Resected Glioblastoma Multiforme Tumors and in an In Vitro Model.

Glioblastoma multiforme (GBM) is a malignant tumor with a higher prevalence in men and a higher survival rate in transmenopausal women. It exhibits distinct areas influenced by changing environmental conditions. This study examines how these areas differ in the levels of estrogen receptors (ERs) which play an important role in the development and progression of many cancers, and whose expression levels are often correlated with patient survival. This study utilized two research models: an in vitro model employing the U87 cell line and a second model involving tumors resected from patients (including tumor core, enhancing tumor region, and peritumoral area). ER expression was assessed at both gene and protein levels, with the results validated using confocal microscopy and immunohistochemistry. Under hypoxic conditions, the U87 line displayed a decrease in ERß mRNA expression and an increase in ERα mRNA expression. In patient samples, ERß mRNA expression was lower in the tumor core compared to the enhancing tumor region (only in males when the study group was divided by sex). In addition, ERß protein expression was lower in the tumor core than in the peritumoral area (only in women when the study group was divided by sex). Immunohistochemical analysis indicated the highest ERß protein expression in the enhancing tumor area, followed by the peritumoral area, and the lowest in the tumor core. The findings suggest that ER expression may significantly influence the development of GBM, exhibiting variability under the influence of conditions present in different tumor areas.

The Severity of Diabetic Retinopathy Corresponds with Corneal Nerve Alterations and Ocular Discomfort of the Patient.

Diabetic retinopathy (DR) remains the leading cause of blindness in the working-age population. Its progression causes gradual damage to corneal nerves, resulting in decreased corneal sensitivity (CS) and disruption of anterior-eye-surface homeostasis, which is clinically manifested by increased ocular discomfort and dry eye disease (DED). This study included 52 DR patients and 52 sex- and age-matched controls. Ocular Surface Disease Index (OSDI) survey, tear film-related parameters, CS, and in vivo corneal confocal microscopy (IVCM) of the subbasal plexus were performed. Furthermore, all patients underwent tear sampling for neurotrophin and cytokine analysis. OSDI scores were greater in DR patients than in controls (p = 0.00020). No differences in the Schirmer test score, noninvasive tear film-break-up time (NIBUT), tear meniscus or interferometry values, bulbar redness, severity of blepharitis or meibomian gland loss were found. In the DR group, both the CS (p < 0.001), and the scotopic pupil diameter (p = 0.00008) decreased. IVCM revealed reduced corneal nerve parameters in DR patients. The stage of DR was positively correlated with the OSDI (Rs = +0.51, 95% CI: + 0.35-+0.64, p < 0.001) and negatively correlated with IVCM corneal nerve parameters and scotopic pupillometry (Rs = -0.26, 95% CI: -0.44--0.06, p = 0.0097). We found negative correlations between the OSDI and IVCM corneal innervation parameters. The DR group showed lower tear film-brain-derived neurotrophic factor (BDNF) levels (p = 0.0001) and no differences in nerve growth factor (NGF)-ß, neurotrophin (NT)-4, vascular endothelial growth factor (VEGF), interleukin (IL)-1ß, IL-4, IL-5, IL-6, or IL-12 concentrations. Tumor necrosis factor (TNF)-α, IL-2, IL-8, IL-10, granulocyte macrophage colony-stimulating factor (GM-CSF), and interferon (IFN)-γ levels were decreased among patients with DR. Corneal innervation defects have a direct impact on patients' subjective feelings. The evolution of DR appears to be associated with corneal nerve alterations, emphasizing the importance of IVCM.

DHEA and Its Metabolites Reduce the Cytokines Involved in the Inflammatory Response and Fibrosis in Primary Biliary Cholangitis.

Dehydroepiandrosterone (DHEA) is an abundant steroid and precursor of sex hormones. During aging, the reduction in DHEA synthesis causes a significant depletion of estrogens and androgens in different organs, such as the ovaries, brain, and liver. Primary Biliary Cholangitis (PBC) is a cholestatic liver disease that begins with immune-mediated bile duct damage, and is followed by liver fibrosis, and finally, cirrhosis. PBC primarily affects postmenopausal women, with an average age of diagnosis of 65 years, but younger women are also affected. Here, we analyzed the levels of DHEA, estradiol (E2), and estriol (E3) in the PBC sera of females at an age of diagnosis under 40 (n = 37) and above 65 (n = 29). Our results indicate that in PBC patients at an age of diagnosis under 40, E2 was significantly lower compared to that in healthy women. In contrast, the levels of DHEA and E3 were in a normal range. Furthermore, ELISA assays revealed that in PBC patients at an age of diagnosis above 65, the levels of DHEA, E2, and E3 significantly declined in comparison to those in younger patients. In addition, flow cytometry analysis showed that the level of IL-8 significantly decreased while the level of TNF-α increased in older PBC patients compared to younger ones. Moreover, we showed for the first time that the sulfonated form of DHEA, DHEA-S, reduces the levels of both pro-inflammatory interleukins, IL-8 and TNF-α, in PBC-like cholangiocytes (H69-miR506), while it diminishes the level of the pro-fibrotic interleukin, IL-13, in hepatocytes (Hep-G2). Finally, we demonstrated that the expression of the pro-fibrotic agent TGF-ß significantly increased in both the early (F0-F3) and cirrhotic (F4) stages of PBC, and this elevation was accompanied by higher α-SMA expression.

Androgen Receptor Expression in the Various Regions of Resected Glioblastoma Multiforme Tumors and in an In Vitro Model.

Glioblastoma multiforme (GBM) is a malignant glioma, difficult to detect and with the lowest survival rates among gliomas. Its greater incidence among men and its higher survival rate among premenopausal women suggest that it may be associated with the levels of androgens. As androgens stimulate the androgen receptor (AR), which acts as a transcription factor, the aim of this study was the investigate the role of AR in the progression of GBM. The study was conducted on tissues collected from three regions of GBM tumors (tumor core, enhancing tumor region, and peritumoral area). In addition, an in vitro experiment was conducted on U-87 cells under various culture conditions (necrotic, hypoxic, and nutrient-deficient), mimicking the conditions in a tumor. In both of the models, androgen receptor expression was determined at the gene and protein levels, and the results were confirmed by confocal microscopy and immunohistochemistry. AR mRNA expression was higher under nutrient-deficient conditions and lower under hypoxic conditions in vitro. However, there were no differences in AR protein expression. No differences in AR mRNA expression were observed between the tested tumor structures taken from patients. No differences in AR mRNA expression were observed between the men and women. However, AR protein expression in tumors resected from patients was higher in the enhancing tumor region and in the peritumoral area than in the tumor core. In women, higher AR expression was observed in the peritumoral area than in the tumor core. AR expression in GBM tumors did not differ significantly between men and women, which suggests that the higher incidence of GBM in men is not associated with AR expression. In the group consisting of men and women, AR expression varied between the regions of the tumor: AR expression was higher in the enhancing tumor region and in the peritumoral area than in the tumor core, showing a dependence on tumor conditions (hypoxia and insufficient nutrient supply).

Electric Cell-Substrate Impedance Sensing (ECIS) as a Convenient Tool to Assess the Potential of Low Molecular Fraction Derived from Medicinal Fungus Cerrena unicolor in Action on L929 and CT-26 Cell Lines.

The increase in the incidence of cancer has contributed to the search for new therapeutic methods. In recent years, the use of preparations of natural origin from medical fungi has increased. One such active substance is the extracellular, low molecular active fraction obtained from the medicinal fungus Cerrena unicolor. This study aimed to monitor the pharmacokinetics of different concentrations of substances isolated from the medicinal fungus Cerrena unicolor (ex-LMS) using the ECIS technique. In the study, mouse L929 fibroblasts and colon cancer CT26 cell lines were treated with different concentrations of the active fractions obtained from Cerrena unicolor: C1 = 2.285 (µg/mL); C2 = 22.85 (µg/mL); and C3 = 228.5 (µg/mL). This study demonstrated that the tested preparation from Cerrena unicolor had no considerable effect on the resistance, capacitance, and impedance of L929 fibroblast cells, which was an indicator of no significant effect on its physiological processes. At the same time, those parameters exhibited a decrease in colon cancer cell viability. Following our previous and current studies on Cerrena unicolor, ex-LMS extracts can be safely used in anticancer therapy or chemoprevention with no significant harmful effects on normal cells.

Garcinol-A Natural Histone Acetyltransferase Inhibitor and New Anti-Cancer Epigenetic Drug.

Garcinol extracted from Garcinia indica fruit peel and leaves is a polyisoprenylated benzophenone. In traditional medicine it was used for its antioxidant and anti-inflammatory properties. Several studies have shown anti-cancer properties of garcinol in cancer cell lines and experimental animal models. Garcinol action in cancer cells is based on its antioxidant and anti-inflammatory properties, but also on its potency to inhibit histone acetyltransferases (HATs). Recent studies indicate that garcinol may also deregulate expression of miRNAs involved in tumour development and progression. This paper focuses on the latest research concerning garcinol as a HAT inhibitor and miRNA deregulator in the development and progression of various cancers. Garcinol may be considered as a candidate for next generation epigenetic drugs, but further studies are needed to establish the precise toxicity, dosages, routes of administration, and safety for patients.

Synthesis and Anticancer Activity of Mitotic-Specific 3,4-Dihydropyridine-2(1H)-thiones.

Most anticancer drugs target mitosis as the most crucial and fragile period of rapidly dividing cancer cells. However the limitations of classical chemotherapeutics drive the search for new more effective and selective compounds. For this purpose structural modifications of the previously characterized pyridine aalog (S1) were incorporated aiming to obtain an antimitotic inhibitor of satisfactory and specific anticancer activity. Structure-activity relationship analysis of the compounds against a panel of cancer cell lines allowed to select a compound with a thiophene ring at C5 of a 3,4-dihydropyridine-2(1H)-thione (S22) with promising antiproliferative activity (IC50 equal 1.71 ± 0.58 µM) and selectivity (SI = 21.09) against melanoma A375 cells. Moreover, all three of the most active compounds from the antiproliferative study, namely S1, S19 and S22 showed better selectivity against A375 cells than reference drug, suggesting their possible lower toxicity and wider therapeutic index. As further study revealed, selected compounds inhibited tubulin polymerization via colchicine binding site in dose dependent manner, leading to aberrant mitotic spindle formation, cell cycle arrest and apoptosis. Summarizing, the current study showed that among obtained mitotic-specific inhibitors analogue with thiophene ring showed the highest antiproliferative activity and selectivity against cancer cells.

Structure-function analysis of ß-arrestin Kurtz reveals a critical role of receptor interactions in downregulation of GPCR signaling in vivo.

Arrestins control signaling via the G protein coupled receptors (GPCRs), serving as both signal terminators and transducers. Previous studies identified several structural elements in arrestins that contribute to their functions as GPCR regulators. However, the importance of these elements in vivo is unclear, and the developmental roles of arrestins are not well understood. We carried out an in vivo structure-function analysis of Kurtz (Krz), the single ortholog of mammalian ß-arrestins in the Drosophila genome. A combination of Krz mutations affecting the GPCR-phosphosensing and receptor core-binding ("finger loop") functions (Krz-KKVL/A) resulted in a complete loss of Krz activity during development. Endosome recruitment and bioluminescence resonance energy transfer (BRET) assays revealed that the KKVL/A mutations abolished the GPCR-binding ability of Krz. We found that the isolated "finger loop" mutation (Krz-VL/A), while having a negligible effect on GPCR internalization, severely affected Krz function, suggesting that tight receptor interactions are necessary for proper termination of signaling in vivo. Genetic analysis as well as live imaging demonstrated that mutations in Krz led to hyperactivity of the GPCR Mist (also known as Mthl1), which is activated by its ligand Folded gastrulation (Fog) and is responsible for cellular contractility and epithelial morphogenesis. Krz mutations affected two developmental events that are under the control of Fog-Mist signaling: gastrulation and morphogenesis of the wing. Overall, our data reveal the functional importance in vivo of direct ß-arrestin/GPCR binding, which is mediated by the recognition of the phosphorylated receptor tail and receptor core interaction. These Krz-GPCR interactions are critical for setting the correct level of Fog-Mist signaling during epithelial morphogenesis.

Synthesis and biological evaluation of 3-functionalized 2-phenyl- and 2-alkylbenzo[b]furans as antiproliferative agents against human melanoma cell line.

The key function of microtubules and mitotic spindle in cell division make them attractive targets in anticancer therapy. In the present study, functionalized in 3 position 2-phenyl- and 2-alkylbenzo[b]furans were synthesized and evaluated as antitumor agents. Among the synthesized derivatives 13a, 13b and 14 exhibited the most potent antiproliferative activity against human melanoma A375 cell line with IC50 values of 2.85 µM, 0.86 µM, 0.09 µM, respectively. The most promising compound defined was 14 with three methoxy groups in the 3-aroyl substituent and 7-methoxy group in 2-phenylbenzo[b]furan skeleton. Tubulin polymerization assay, confocal microscopy imaging and flow cytometry analysis revealed that 2-phenyl-3-aroylbenzo[b]furans (13a, 13b and 14) inhibited tubulin polymerization leading to disruption of mitotic spindle formation, cell cycle arrest in G2/M phase and apoptosis.

Membrane Transporters in Human Parotid Gland-Targeted Proteomics Approach.

Salivary glands provide secretory functions, including secretion of xenobiotics and among them drugs. However, there is no published information about protein abundance of drug transporters measured using reliable protein quantification methods. Therefore, mRNA expression and absolute protein content of clinically relevant ABC (n = 6) and SLC (n = 15) family member transporters in the human parotid gland, using the qRT-PCR and liquid chromatography‒tandem mass spectrometry (LC-MS/MS) method, were studied. The abundance of nearly all measured proteins ranged between 0.04 and 0.45 pmol/mg (OCT3 > MRP1 > PEPT2 > MRP4 > MATE1 > BCRP). mRNAs of ABCB1, ABCC2, ABCC3, SLC10A1, SLC10A2, SLC22A1, SLC22A5, SLC22A6, SLC22A7, SLC22A8, SLCO1A2, SLCO1B1, SLCO1B3 and SLCO2B1 were not detected. The present study provides, for the first time, information about the protein abundance of membrane transporters in the human parotid gland, which could further be used to define salivary bidirectional transport (absorption and secretion) mechanisms of endogenous compounds and xenobiotics.

Associations of microRNAs, Angiogenesis-Regulating Factors and CFH Y402H Polymorphism-An Attempt to Search for Systemic Biomarkers in Age-Related Macular Degeneration.

Age-related macular degeneration (AMD) remains the leading cause of blindness in elderly people, but the pathophysiology of this disease is still largely unknown. We investigated the systemic expression of angiogenesis-regulating growth factors and selected miRNAs known to regulate angiogenesis in AMD patients. We also focused on possible correlations of their expression with the presence of CFH Y402H or ARMS A69S risk variants. A total of 354 AMD patients and 121 controls were enrolled in this study. The levels of angiogenesis-regulating factors were analyzed in plasma samples using Luminex technology. The expression of selected miRNAs was analyzed in peripheral blood plasma using real-time qPCR. The genetic analysis was performed with an Illumina NextSeq500 system. AMD was an independent factor associated with lower levels of angiogenin (ß = -0.29, p < 0.001), endostatin (ß = -0.18, p < 0.001), FGF-basic (ß = -0.18, p < 0.001), PlGF (ß = -0.24, p < 0.001), miRNA-21-3p (ß = -0.13, p = 0.01) and miRNA-155-5p (ß = -0.16, p = 0.002); and with higher levels of FGF-acidic (ß = 0.11, p = 0.03), miRNA-23a-3p (ß = 0.17, p < 0.001), miRNA-126-5p (ß = 0.13, p = 0.009), miRNA-16-5p (ß = 0.40, p < 0.001), miRNA-17-3p (ß = 0.13, p = 0.01), miRNA-17-5p (ß = 0.17, p < 0.001), miRNA-223-3p (ß = 0.15, p = 0.004), and miRNA-93 (ß = 0.11, p = 0.04). The expression of analyzed miRNA molecules significantly correlated with the levels of tested angiogenesis-regulating factors and clinical parameters in AMD patients, whereas such correlations were not observed in controls. We also found an association between the CFH Y402H polymorphism and miRNA profiles, whereby TT homozygotes showed evidently higher expression of miRNA-16-5p than CC homozygotes or TC heterozygotes (p = 0.0007). Our results suggest that the balance between systemic pro- and anti-angiogenic factors and miRNAs is vital in multifactorial AMD pathogenesis.

Mutations in the profilin 1 gene cause familial amyotrophic lateral sclerosis.

Amyotrophic lateral sclerosis (ALS) is a late-onset neurodegenerative disorder resulting from motor neuron death. Approximately 10% of cases are familial (FALS), typically with a dominant inheritance mode. Despite numerous advances in recent years, nearly 50% of FALS cases have unknown genetic aetiology. Here we show that mutations within the profilin 1 (PFN1) gene can cause FALS. PFN1 is crucial for the conversion of monomeric (G)-actin to filamentous (F)-actin. Exome sequencing of two large ALS families showed different mutations within the PFN1 gene. Further sequence analysis identified 4 mutations in 7 out of 274 FALS cases. Cells expressing PFN1 mutants contain ubiquitinated, insoluble aggregates that in many cases contain the ALS-associated protein TDP-43. PFN1 mutants also display decreased bound actin levels and can inhibit axon outgrowth. Furthermore, primary motor neurons expressing mutant PFN1 display smaller growth cones with a reduced F/G-actin ratio. These observations further document that cytoskeletal pathway alterations contribute to ALS pathogenesis.

The macular function and structure in patients with diabetic macular edema before and after ranibizumab treatment.

PURPOSE: To evaluate macular function and structure in patients with diabetic macular edema prior to, as well as 3 and 6 months after intravitreal ranibizumab treatment. PATIENTS AND METHODS: Seventeen eyes of 17 patients with type 2 diabetes mellitus and diabetic macular edema (DME) were treated with intravitreal injections of 0.5 mg ranibizumab. Prior to the first injection, as well as after 3 and 6 months, the following examinations were performed: assessment of distance best-corrected visual acuity (log MAR), perception of metamorphopsia (M-Chart), slit lamp examination of the anterior and posterior segment of the eye (Volk 90D lens), evaluation of the retinal and choroidal circulation (fluorescein angiography), assessment of the structure and thickness of the macula (OCT), as well as evaluation of the macular function (PERG and mfERG). RESULTS: We observed that ranibizumab significantly improved visual acuity after 3 and 6 months from the beginning of the treatment, which was a consequence of reduced macular edema and vascular leakage. There was a statistically significant decrease in metamorphopsia frequency at month 3; however, at month 6 it was a statistically insignificant when compared to the baseline. The results of electrophysiological examinations revealed no improvement in ranibizumab-treated patients. CONCLUSION: Improvement of visual acuity and reduction in macular thickness were maintained up to the 6-month follow-up. The results of electrophysiological examinations revealed that ranibizumab injections tend to stabilize bioelectrical macular function of the outer, middle and inner retinal layers, which was impossible to recognize on the basis of visual acuity and OCT. Therefore, the electrophysiological examinations should be used as an additional objective tool for the evaluation of the anti-VEGF treatment effectiveness in DME.

Aqueous levels of VEGF correlate with retinal non-perfusion areas in patients with diabetic macular edema and macular edema secondary to central retinal vein occlusion.

AIM: To evaluate the association between the level of vascular endothelial growth factor in the aqueous humor and the size of capillary non-perfusion areas in patients with macular edema secondary to retinal vein occlusion and diabetic retinopathy. MATERIAL AND METHODS: The study group consisted of 24 patients (24 eyes) at the age of 55-78 years, with diffuse macular edema secondary to retinal vein occlusion and diabetic retinopathy. The control group consisted of 26 subjects aged 55-87 years who were admitted for scheduled cataract surgery. The VEGF aqueous humor levels, retinal thickness using optical coherence tomography, as well as the size of non-perfusion areas measured on fluorescein angiography images were evaluated in each enrolled subject. RESULTS: The vascular endothelial growth factor aqueous humor levels were found to be significantly higher in patients with macular edema as compared to controls (p = 0.0002). In the diabetic macular edema and retinal vein occlusion group, the con- centration of vascular endothelial growth factor in aqueous humor positively correlated with the extent of non-perfusion areas measured on fluorescein angiograms (Rs = + 0.45, p = 0.02;). Multivariate analysis of patients and controls performed using the general linear model, adjusted for age, sex, intraocular pressure and the presence of diabetes, revealed that macular edema was an independent factor associated with higher aqueous VEGF concentrations (ß = +0.74, p = 0.0012). CONCLUSIONS: Macular edema secondary to either retinal vein occlusion or diabetic retinopathy is associated with the increased levels of vascular endothelial growth factor in the aqueous humor. Therefore, the management of patients with macular edema secondary to retinal vein occlusion or diabetic retinopathy should aim at reducing the ocular vascular endothelial growth factor concentrations, especially in the presence of capillary non-perfusion areas.

The Effects of Cadmium at Low Environmental Concentrations on THP-1 Macrophage Apoptosis.

Cadmium at environmental concentrations is a risk factor for many diseases, including cardiovascular and neurodegenerative diseases, in which macrophages play an important role. The aim of this study was to evaluate the effects of cadmium at low environmental (nanomolar) concentrations on apoptotic processes in THP-1(acute monocytic leukemia cells line)-derived macrophages, with special focus on mitochondrial events involved. Macrophages were incubated with various cadmium chloride (CdCl2) solutions for 48 h at final concentrations of 5 nM, 20 nM, 200 nM and 2 µM CdCl2. Cell viability was measured using flow cytometry. Flow cytometric measurement (annexin V/FITC (annexin V/fluorescein isothiocyanate) and PI (propidium iodide) double staining) was used to quantify the extent of apoptosis. Fluorescence and confocal microscopy were used for imaging of apoptosis process. Changes in mitochondrial membrane potential were monitored using cytofluorimetry after cell staining with JC-1(5,5',6,6'-tetrachloro-1,1',3,3'-tetraethylbenzimidazol-carbocyane iodide) probe. Mitochondrial ROS (reactive oxygen species) levels were measured cytofluorimetrically after incubation of cells with mitochondrial superoxide indicator (MitoSOX) red fluorescent marker. The mRNA expression of Bcl-2 and Bax was analysed with qRT-PCR. Our study demonstrates that cadmium, even at low environmental concentrations, exerts mitochondrial toxicity in THP-1 macrophages. Forty-eight-hour exposure to very low concentrations reduces cell viability and results in cell death by apoptosis and necrosis. The decrease in mitochondrial membrane potential, increased ROS production, increased Bax and decreased Bcl-2 mRNA expression are mitochondrial events involved in cadmium-induced apoptosis.

Evidence that the population of quiescent bone marrow-residing very small embryonic/epiblast-like stem cells (VSELs) expands in response to neurotoxic treatment.

The concept that bone marrow (BM)-derived cells may participate in neural regeneration remains controversial, and the identity of the specific cell type(s) involved remains unknown. We recently reported that the adult murine BM contains a highly mobile population of Sca-1(+) Lin(-) CD45(-) cells known as very small embryonic/epiblast-like stem cells (VSELs) that express several markers of pluripotency such as Oct-4. In the BM microenvironment, these cells are kept quiescent because of epigenetic modification of certain paternally imprinted genes. However, as reported, these cells can be mobilized in mice in an experimental model of stroke and express several genes involved in neurogenesis while circulating in peripheral blood (PB). In the current work, we employed a model of toxic brain damage, which is induced by administration of kainic acid, to see not only whether VSELs can be mobilized into PB in response to this neurotoxin, but, more importantly, whether they proliferate and expand in BM tissue. We report here for the first time that brain damage leads to activation and expansion of the BM pool of quiescent VSELs, which precedes their subsequent egress into PB. Harnessing these cells in neural tissue regeneration is currently one of the challenges in regenerative medicine.

Perinatal mortality in urban and rural areas in Poland in 2002-2012.

AIM: The aim of this study was to analyze the level and trends of perinatal mortality by mother's place of residence (urban vs rural area) in Poland in 2002-2012. MATERIAL AND METHOD: This study was based on the data of the Central Statistical Office on the number of live births, infant deaths (0-6 days) and stillbirths by mother's place of residence (urban vs rural area), reported in 2002-2012 in 16 provinces and Poland in general. Joinpoint model was used to analyze perinatal mortality rate trends over time and average annual percent change (APC). Urban/rural ratio was employed to demonstrate the differences in perinatal mortality between urban and rural areas. RESULTS: In the period analyzed, perinatal mortality in Poland decreased by 3.4% (p<0.05) and 2.7% (p<0.05) per year in urban and rural areas, respectively. Having considered urban areas, perinatal mortality rate was decreasing at the fastest pace in the following provinces: Pomorskie (APC) = -6.6%, p<0.05), Warminsko-Mazurskie (APC) = -5.4%, p<0.05), Lubuskie i Swietokrzyskie (APC = -4.5%, p<0.05) while for rural areas - Dolnoslaskie (APC = -4.3%, p<0.05), Wielkopolskie, Zachodniopomorskie (APC = -3.7%, p<0.05) and Slaskie (APC = -3.2%, p<0.05). CONCLUSIONS: In the study period, a decrease in perinatal mortality was reported in Poland, both in urban and rural areas. The level of perinatal mortality rate as well as the pace of these changes differed between provinces.

[Increased expression of endothelin-1-- a novel diagnostic marker for early AMD detection?]. / Wzrost ekspresji endoteliny-1--nowy marker wczesnej diagnostyki AMD?

AIM: The relationship between ischemic vascular disease and age-related macular degeneration may indicate the role of vascular injury as the primary insult causing functional deficits in age-related macular degeneration. The vasoactive factors produced by endothelial cells include endothelin-1 (ET-1), which is one of the most potent vasoconstricting peptides. In this study we sought to explore the potential role of endothelial dysfunction in the pathogenesis of age-related macular degeneration by measuring the concentration of ET-1 in peripheral blood of individuals diagnosed with age-related macular degeneration and evaluating its intracellular expression in peripheral blood cells, on mRNA level. MATERIAL AND METHODS: Peripheral blood samples from 31 patients with diagnosed dry age-related macular degeneration and 46 patients with neovascular age-related macular degeneration were collected. Forty six age- and sex-matched volunteers without age-related macular degeneration were enrolled as a control group. ET-1 plasma levels were analyzed by ELISA and intracellular expression of ET-1 in peripheral blood cells was studied by using qRT-PCR. RESULTS: The expression of intracellular ET-1 was significantly elevated in peripheral blood cells of both dry and wet age-related macular degeneration patients compared with the control subjects. Immunofluorescence staining revealed that ET-1 was specifically expressed in the circulating endothelial cells. CONCLUSIONS: We assume that damaged endothelial cells may release a variety of vasoconstricting molecules, including ET-1, leading to derangement between the endothelium-derived relaxing and contracting factors. Local retinal ischemia consequently develops which may promote the development of retinal degeneration in patients with age-related macular degeneration,

[Idiopathic choroidal neovascularization--case report]. / Idiopatyczna neowaskularyzacja podsiatkówkowa--opis przypadku.

PURPOSE: To report a patient with peripapillary idiopathic choroidal neovascularization treated with an anti-VEGF (anti- vascular endothelial growth factor) agent (bevacizumab) observed during for 9 months. PATIENT AND METHODS: Twenty nine years old man was referred to the Department for diagnosis and treatment because of unilateral visual acuity decrease in the right eye (VA RE--0.1) and metamorphopsias. The routine ophthalmic examination revealed macular edema and peripapillaryedema with epiretinal and intraretinal hemorrhages. The optical coherence tomography, fluorescein angiography, as well as laboratory tests were performed in order to exclude uveitis. Due to the difficulties in the diagnosis indocyanine green angiography was also performed. Based on clinical symptoms and the findings of the additional diagnostic procedures, the patient was diagnosed with idiopathic choroidal neovascularization. The patient was qualified for anti-VEGF therapy and received three intravitreal injections of bevacizumab at the dose of 1.25 mg, at monthly intervals. RESULTS: Significant improvement of visual acuity (VA RE - 1.0) and regression of the peripapillary edema with hemorrhages were achieved after the third injection of 1.25 mg bevacizumab. At 6 months, peripapillary scarring was observed in the area involved by the primary lesions. CONCLUSIONS: Anti-VEGF therapy is the effective treatment of idiopathic choroidal neovascularization in the described case. The visual acuity improvement and rapid regression of posterior segment lesions after bevacizumab administration were observed.

Effect of R&D activity on level of innovation of EU countries in 2014 and 2020.

Research and development carried out by companies are pivotal for innovative economies of countries, especially in the unpredictable and changing social, economic and political environment. In this context, it is very important to answer, which factors identify the effectiveness of measures in relation to R&D activity and innovativeness in EU countries and how should the degree of development of a country be assessed in terms of R&D activity? The purpose of this article is to verify level of innovativeness and degree of research and development (R&D) activity in EU countries in the years 2014 and 2020 using Hellwig's measure of development. To achieve this, qualitative and quantitative analysis, synthesis, deduction and induction, comparative analysis, and reasoning by analogy of phenomena were employed. The research was conducted on the basis of the expertly selected variables for their relation to R&D activity from a number of sources, such as Eurostat, World Bank Data, etc.. The indicated variables were analysed using statistical methods and then subjected to a linear ordering procedure based on the Hellwig development pattern method. Thanks to the research results, it is possible to indicate areas in which the initiation of activities would have the greatest degree of influence on development of R&D activity, thus influencing the increase in the level of innovativeness of a country. Indicators relating to R&D activity were selected and then used as variables to study the effect of the degree of R&D activity in EU countries in the years 2014 and 2020 on the level of innovativeness of these countries. The conducted research coincides with the results presented in the European Innovation Scoreboard. There is a significant correlation between the development of R&D activities and innovation performance.