Morphological evaluation of the feline placenta correlates with gene expression of vascular growth factors and receptors†.

Placental angiogenesis is critical for normal development. Angiogenic factors and their receptors are key regulators of this process. Dysregulated placental vascular development is associated with pregnancy complications. Despite their importance, vascular growth factor expression has not been thoroughly correlated with placental morphologic development across gestation in cats. We postulate that changes in placental vessel morphology can be appreciated as consequences of dynamic expression of angiogenic signaling agents. Here, we characterized changes in placental morphology alongside expression analysis of angiogenic factor splice variants and receptors throughout pregnancy in domestic shorthair cats. We observed increased vascular and lamellar density in the lamellar zone during mid-pregnancy. Immunohistochemical analysis localized the vascular endothelial growth factor A (VEGF-A) receptor KDR to endothelial cells of the maternal and fetal microvasculatures. PlGF and its principal receptor Flt-1 were localized to the trophoblasts and fetal vasculature. VEGF-A was found in trophoblast cells and associated with endothelial cells. We detected expression of two Plgf splice variants and four Vegf-a variants. Quantitative real-time polymerase chain reaction analysis showed upregulation of mRNAs encoding pan Vegf-a and all Vegf-a splice forms at gestational days 30-35. Vegf-A showed a marked relative increase in expression during mid-pregnancy, consistent with the pro-angiogenic changes seen in the lamellar zone at days 30-35. Flt-1 was upregulated during late pregnancy. Plgf variants showed stable expression during the first two-thirds of pregnancy, followed by a marked increase toward term. These findings revealed specific spatiotemporal expression patterns of VEGF-A family members consistent with pivotal roles during normal placental development.

Expression of Markers Ki-67, Nestin, VEGF, CD34 and Apoptosis in Relatively Healthy Lung Tissue with Non-Changed and Metaplastic Bronchial Epithelium.

BACKGROUND: Knowledge about the occurrence of processes such as proliferation, apoptosis and angiogenesis in healthy lung tissues with different bronchial epitheliums is limited, and further exploration can contribute to a better understanding of the physiological renewal of lung tissues. The processes mentioned above occur with the help of important tissue factors; therefore, the aim of the study was to determine the expression of markers Ki-67, nestin, CD34 and vascular endothelial growth factor (VEFG) and detect apoptotic cells in relatively healthy lung tissue. METHODS: Samples of relatively healthy lung tissue were obtained from 19 patients and divided into groups of patients with non-changed and patients with metaplastic bronchial epithelium. Tissue samples were examined by hematoxylin and eosin staining. Ki-67, nestin, VEGF and CD34-positive cells were detected by the immunohistochemistry method. Terminal deoxynucleotidyl transferase (TdT) dUTP nick-end labeling (TUNEL) assay was carried out to detect apoptotic cells. The number of positive structures was counted semi-quantitatively by microscopy. RESULTS: Ki-67-positive cells were detected in only one case. An occasional to moderate number of nestin-positive structures was found in various tissues of relatively healthy lungs with different bronchial epitheliums. No apoptotic cells were seen in non-changed bronchial epithelium, compared with few apoptotic cells in metaplastic bronchial epithelium. Metaplastic bronchial epithelium contained more VEGF-positive cells than non-changed bronchial epithelium. Samples with non-changed, and metaplastic bronchial epithelium both contained a similar number of CD34-positive structures. CONCLUSIONS: Proliferative activity and programmed cell death are not prominent events in normal lung tissue. A moderate number of nestin-positive cells in the alveolar epithelium and cartilage of bronchi with pseudostratified ciliated epithelium suggests a significant role of neuronal origin cells in these structures, to be intensified in metaplastic bronchial epithelium. A practically non-changed number of CD34-positive cells excludes any difference in stimulation of endothelial origin cells between lungs with different types of epithelium, while an increase in VEGF in structures with metaplastic epithelium suggests the presence/influence of tissue ischemia impact on possible development/maintenance of metaplasia.

sRAGE downregulates the VEGF expression in OHSS ovarian granulosa cells.

OBJECTIVE: Ovarian hyperstimulation syndrome (OHSS) is mainly caused by human chorionic gonadotropin (hCG) through vasoactive mediators such as vascular endothelial growth factor (VEGF) and various inflammatory factors. Our previous study showed that soluble receptor for advanced glycation end products (sRAGE) played a protective role in PCOS by inhibiting VEGF, so wanted to explore the role of sRAGE in OHSS. METHODS: Two sets of experiments were performed in this study. In part one, sRAGE protein levels in follicular fluid (FF) samples from 60 patients with OHSS and 60 non-OHSS patients were measured by ELISA. In part two, ovarian granulosa cells were isolated from an additional 25 patients with OHSS and cultured. Then, ovarian granulosa cells were treated with different concentrations of sRAGE. Granulosa cells cultured without sRAGE stimulation were used as the control group. The levels of VEGF, amphiregulin (AREG), betacellulin (BTC), and epiregulin (EREG) mRNA were examined by quantitative RT-PCR. The protein levels of VEGF, AREG, BTC, and EREG were measured by ELISA. RESULTS: Compared with non-OHSS patients, patients with OHSS exhibited lower sRAGE levels in both serum and FF (p < .05). Treatment with sRAGE decreased the production of VEGF, and the effects were dependent on the concentration of sRAGE (p < .05). Simultaneously, the expression of the EGF-like growth factors AREG, BTC and EREG was decreased, and their expression was dependent on the concentration of sRAGE (p < .05). CONCLUSIONS: sRAGE downregulate VEGF expression in OHSS ovarian granulosa cells, in which EGF-like growth factor pathway may be involved, and sRAGE may play a potential protective role in OHSS.

Clearance of inflammatory cytokines in patients with septic acute kidney injury during renal replacement therapy using the EMiC2 filter (Clic-AKI study).

BACKGROUND: The EMiC2 membrane is a medium cut-off haemofilter (45 kiloDalton). Little is known regarding its efficacy in eliminating medium-sized cytokines in sepsis. This study aimed to explore the effects of continuous veno-venous haemodialysis (CVVHD) using the EMiC2 filter on cytokine clearance. METHODS: This was a prospective observational study conducted in critically ill patients with sepsis and acute kidney injury requiring kidney replacement therapy. We measured concentrations of 12 cytokines [Interleukin (IL) IL-1ß, IL-1α, IL-2, IL-4, IL-6, IL-8, IL-10, interferon (IFN)-γ, tumour necrosis factor (TNF)-α, vascular endothelial growth factor, monocyte chemoattractant protein (MCP)-1, epidermal growth factor (EGF)] in plasma at baseline (T0) and pre- and post-dialyzer at 1, 6, 24, and 48 h after CVVHD initiation and in the effluent fluid at corresponding time points. Outcomes were the effluent and adsorptive clearance rates, mass balances, and changes in serial serum concentrations. RESULTS: Twelve patients were included in the final analysis. All cytokines except EGF concentrations declined over 48 h (p < 0.001). The effluent clearance rates were variable and ranged from negligible values for IL-2, IFN-γ, IL-1α, IL-1ß, and EGF, to 19.0 ml/min for TNF-α. Negative or minimal adsorption was observed. The effluent and adsorptive clearance rates remained steady over time. The percentage of cytokine removal was low for most cytokines throughout the 48-h period. CONCLUSION: EMiC2-CVVHD achieved modest removal of most cytokines and demonstrated small to no adsorptive capacity despite a decline in plasma cytokine concentrations. This suggests that changes in plasma cytokine concentrations may not be solely influenced by extracorporeal removal. TRIAL REGISTRATION: NCT03231748, registered on 27th July 2017.

Significance of vascular endothelium growth factor testing in exhaled breath condensate of patients with acute respiratory distress syndrome.

OBJECTIVE: We aimed to observe and investigate the clinical significance of vascular endothelium growth factor (VEGF) levels in exhaled breath condensate (EBC) from patients with acute respiratory distress syndrome (ARDS). METHODS: An improved EcoScreen condenser was used to collect EBC from 31 ARDS patients on mechanical ventilation and from 22 healthy subjects. Serum and EBC VEGF levels were analyzed with ELISA. VEGF levels in the EBC of patients with different grades of lung injuries were analyzed. The correlation between VEGF levels and clinical indicators was analyzed. RESULTS: Serum and EBC VEGF levels were linearly and positively correlated with a correlation coefficient of 0.694 (P< 0.01). The VEGF level in the EBC of ARDS patients was significantly lower than that in the control group (P< 0.01). The VEGF level in the EBC of the mild ARDS group was higher than that in the moderate-severe ARDS group (P< 0.01). The VEGF level in the EBC of the survival group was higher than that in the mortality group. The VEGF level in the EBC of ARDS patients was positively correlated with PaO2/FiO2 and PaO2 and was negatively correlated with lung injury score (LIS) and A-aDO2/PaO2. CONCLUSION: The changes in VEGF levels in the EBC of ARDS patients can Respiratory Medicine, reflect the severity of lung injury. Therefore, VEGF level in EBC can be used as an auxiliary index for judging the severity and prognosis of ARDS patients.

A modular 3D printed lab-on-a-chip for early cancer detection.

A functional polymeric 3D device is produced in a single step printing process using a stereolithography based 3D printer. The photocurable formulation is designed for introducing a controlled amount of carboxyl groups (-COOH), in order to perform a covalent immobilization of bioreceptors on the device. The effectiveness of the application is demonstrated by performing an immunoassay for the detection of protein biomarkers involved in angiogenesis, whose role is crucial in the onset of cancer and in the progressive metastatic behavior of tumors. The detection of angiogenesis biomarkers is necessary for an early diagnosis of the pathology, allowing the employment of a less invasive therapy for the patient. In particular, vascular endothelial growth factor and angiopoietin-2 biomarkers are detected with a limit of detection of 11 ng mL-1 and 0.8 ng mL-1, respectively. This study shows how 3D microfabrication techniques, material characterization, and device development could be combined to obtain an engineered polymeric chip with intrinsic tuned functionalities.

Utilidad de los marcadores bioquímicos de preeclampsia / Usefulness of the biochemical markers of pre-eclampsia

La preeclampsia (PE) constituye una de las principales causas de mortalidad materna y perinatal en el mundo. En los países desarrollados, los estudios apuntan a un importante aumento de la incidencia de PE en la última década, en parte, por el aumento de la prevalencia, en la población general, de enfermedades que afectan a la función vascular, como la diabetes, la hipertensión crónica o la enfermedad renal. En el presente documento se lleva cabo una revisión actualizada de la PE. Se describen los criterios diagnósticos y la fisiopatología de la enfermedad. El objetivo principal del documento es revisar los nuevos marcadores bioquímicos que pueden ser de utilidad en la práctica clínica para la predicción y el diagnóstico de la PE, así como los distintos métodos mediante los cuales se puede llevar a cabo su determinación

Pre-eclampsia (PE) is one of the leading causes of maternal and perinatal mortality in the world. In developed countries, studies point to a significant increase in the incidence of PE in the last decade, partly due to the increase in the prevalence in the general population of diseases that affect vascular function, such as diabetes. chronic hypertension, or kidney disease. An updated review of PE is presented in this article. The diagnostic criteria and the pathophysiology of the disease are described. The main objective of the document is to review the new biochemical markers that may be useful in clinical practice for the prediction and diagnosis of PE, as well as the different methods by which yey can be determined

High-sensitive immunosensing of protein biomarker based on interfacial recognition-induced homogeneous exponential transcription.

A novel and versatile immunosensing strategy was developed for ultrasensitive and specific detection of proteins by organically integrating interfacial specific target recognition and homogeneous transcription amplification. In principle, classic antigen-antibody sandwich structure on the microplate could realize the specific identification of target protein. Biotinylated DNA probe was subsequently introduced by streptavidin-biotin system as a bridge linking interfacial and homogeneous reaction. The biotinylated DNA initiated exponential transcription amplification in the solution, which converted per target recognition event on the interface to numerous single-stranded RNA products in solution for highly sensitive fluorescence immunosensing. The proposed immunoassay based on interfacial recognition-induced homogeneous exponential transcription (IR-HET) for vascular endothelial growth factor (VEGF) detection showed a good linear range from 0.01 to 1000 pg/mL and the limit of detection as low as 1 fg/mL, which was 3 orders lower than traditional ELISA method. The established strategy was also successfully applied to directly detect VEGF from culture supernatants of tumor cells and clinical body fluid samples, proving very high sensitivity, selectivity and low matrix effect. Therefore, IR-HET-based immunosensing strategy might become a potential powerful tool be applied in ultrasensitive detection of low abundance protein biomarker for clinical early diagnosis, treatment and prognosis.

Diagnóstico de síndrome de POEMS tras neuropatía de larga evolución / Diagnosis of POEMS syndrome in a patient with long-standing neuropathy

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In Vivo Choroidal Neovascularization and Macrophage Studies Provide Further Evidence for a Broad Role of Prostacyclin in Angiogenesis.

PURPOSE: The purpose of these studies was (1) to investigate the ability of human M1 phenotype macrophages to secrete vascular endothelial growth factor (VEGF) and the influence of prostacyclin receptor (IP) stimulation (2) to evaluate the contribution of the proangiogenic prostanoid prostacyclin to experimental choroidal neovascularization Methods: Human macrophages derived from primary blood mononuclear cells were functionally biased toward the M1 phenotype by using tumor necrosis factor α (TNFα). Experimental choroidal neovascularization was produced by laser photocoagulation. Antagonist drugs RO-3244794 (IP antagonist) and GW 627368 (EP4 antagonist) were administered according to an optimal dosing regimen that was predetermined by bioavailability studies. RESULTS: IP receptor stimulation had diametrically opposed effects on VEGF release compared with reported data on cytokine/chemokine secretion from human macrophages. For example, the IP agonist cicaprost stimulated VEGF secretion although it inhibits monocyte chemoattractant protein-1 (MCP-1) secretion: both would favor a proangiogenic effect. The IP receptor antagonist RO-3244794 produced an ∼20% statistically significant reduction in the neovascularized lesion area in the choroidal neovascularization model, which was a similar level to that produced by the EP4 antagonist GW 627368. Combining the 2 drugs produced a statistically significant reduction in neovascularization but only of slightly greater magnitude than that obtained with each antagonist administered alone. CONCLUSIONS: IP receptor stimulation potently and highly efficaciously promoted VEGF release from human M1 macrophages, indicating a possible contribution of the M1 macrophage subtype to VEGF-induced choroidal neovascularization. Studies in living animals suggest that prostacyclin and its target IP receptor contribute to choroidal neovascularization, although to a more modest extent than might have been expected.

Oral implant osseointegration model in C57Bl/6 mice: microtomographic, histological, histomorphometric and molecular characterization.

INTRODUCTION: Despite the successful clinical application of titanium (Ti) as a biomaterial, the exact cellular and molecular mechanisms responsible for Ti osseointegration remains unclear, especially because of the limited methodological tools available in this field. OBJECTIVE: In this study, we present a microscopic and molecular characterization of an oral implant osseointegration model using C57Bl/6 mice. MATERIAL AND METHODS: Forty-eight male wild-type mice received a Ti implant on the edentulous alveolar crest and the peri-implant sites were evaluated through microscopic (µCT, histological and birefringence) and molecular (RealTimePCRarray) analysis in different points in time after surgery (3, 7, 14 and 21 days). RESULTS: The early stages of osseointegration were marked by an increased expression of growth factors and MSC markers. Subsequently, a provisional granulation tissue was formed, with high expression of VEGFb and earlier osteogenic markers (BMPs, ALP and Runx2). The immune/inflammatory phase was evidenced by an increased density of inflammatory cells, and high expression of cytokines (TNF, IL6, IL1) chemokines (CXCL3, CCL2, CCL5 and CXC3CL1) and chemokine receptors (CCR2 and CCR5). Also, iNOS expression remained low, while ARG1 was upregulated, indicating predominance of a M2-type response. At later points in time, the bone matrix density and volume were increased, in agreement with a high expression of Col1a1 and Col21a2. The remodelling process was marked by peaks of MMPs, RANKL and OPG expression at 14 days, and an increased density of osteoclasts. At 21 days, intimate Ti/bone contact was observed, with expression of final osteoblast differentiation markers (PHEX, SOST), as well as red spectrum collagen fibers. CONCLUSIONS: This study demonstrated a unique molecular view of oral osseointegration kinetics in C57Bl/6 mice, evidencing potential elements responsible for orchestrating cell migration, proliferation, ECM deposition and maturation, angiogenesis, bone formation and remodeling at the bone-implant interface in parallel with a novel microscopic analysis.

Molecularly Imprinted Nanocavities Capable of Ligand-Binding Domain and Size/Shape Recognition for Selective Discrimination of Vascular Endothelial Growth Factor Isoforms.

Vascular endothelial growth factor 165 (VEGF165) is known to be predominantly expressed in the first stage of vascularization; therefore, the detection of VEGF165 is important in the stage diagnosis of cancers. Molecularly imprinted nanocavities, capable of the selective discrimination of VEGF165 from other VEGF isoforms, were prepared by surface-initiated atom transfer radical polymerization. VEGF165 was immobilized on a gold-coated glass substrate by anchored heparin moieties, where the immobilized heparin was able to capture VEGF165 by binding with the heparin-binding domain (HBD) on VEGF165. Molecular imprinting was conducted on the immobilized VEGF165 by using methacrylic acid (MAA) as a functional monomer to interact with basic amino acids outside of the HBD of VEGF165 by electrostatic interaction. After the removal of VEGF165 from the obtained polymer thin layer (ca. 7 nm), VEGF165-imprinted nanocavities remained, in which the heparin moiety and MAA residues were located in suitable positions for VEGF165 recognition. The molecularly imprinted polymer (MIP) thin layer showed a binding affinity for VEGF165 (dissociation constant: 3.4 nM) that was ten times higher than that of the substrate before polymerization (heparin-immobilized substrate). A much lower binding affinity for VEGF121, which contains no heparin-binding domain, was observed. Moreover, the MIP thin layer distinguished VEGF165 from VEGF189, which possesses a larger molecular size than VEGF165, an amino acid sequence homology of 87%, and contains HBDs, whereas the heparin-immobilized substrate showed almost no selectivity. These results suggested that the heparin moiety within the nanocavity provided HBD selectivity and the polymer matrix composed of the molecularly imprinted nanocavity provided size/shape selectivity, which resulted in the highly selective discrimination of VEGF isoforms.

Colorimetric immunoassays for the screening and specificity evaluation of molecules disturbing VEGFs/VEGFRs interactions.

Angiogenesis and its involved proteins, particularly Vascular Endothelial Growth Factor family (VEGFs) and VEGF receptors (VEGFRs), have been considered as a target of therapeutic interest for numerous inflammatory and vascular diseases. Acting on this biological process through interaction with VEGFs or VEGFRs has received considerable attention. Indeed, VEGFs and VEGFRs are currently targeted by drugs such as monoclonal antibodies. The feasibility of a therapeutic strategy based on blocking the VEGF/VEGFR interaction by using ligands "other-than-biologics" is also explored. To help to the discovery of new molecules, screening assays have been developed, particularly to evaluate the VEGFA/VEGFR1 interaction. Despite the therapeutic importance of VEGFB and PlGF (Placental Growth Factor), no assays have been developed to evaluate molecules against their interactions with VEGFR1. Here, we present new versatile colorimetric immunoassays to screen and evaluate the specific interaction of discovered molecules with different growth factors (VEGFA, VEGFB, PlGF) and receptors (VEGFR1, VEGFR2). These tests, based on competitive immunoassay format, will provide essential information on specificity and selectivity of molecules for their targets and will help to work on the pharmaco-modulation of molecules for targeting one specific interaction.

Oral implant osseointegration model in C57Bl/6 mice: microtomographic, histological, histomorphometric and molecular characterization

Abstract Despite the successful clinical application of titanium (Ti) as a biomaterial, the exact cellular and molecular mechanisms responsible for Ti osseointegration remains unclear, especially because of the limited methodological tools available in this field. Objective: In this study, we present a microscopic and molecular characterization of an oral implant osseointegration model using C57Bl/6 mice. Material and Methods: Forty-eight male wild-type mice received a Ti implant on the edentulous alveolar crest and the peri-implant sites were evaluated through microscopic (μCT, histological and birefringence) and molecular (RealTimePCRarray) analysis in different points in time after surgery (3, 7, 14 and 21 days). Results: The early stages of osseointegration were marked by an increased expression of growth factors and MSC markers. Subsequently, a provisional granulation tissue was formed, with high expression of VEGFb and earlier osteogenic markers (BMPs, ALP and Runx2). The immune/inflammatory phase was evidenced by an increased density of inflammatory cells, and high expression of cytokines (TNF, IL6, IL1) chemokines (CXCL3, CCL2, CCL5 and CXC3CL1) and chemokine receptors (CCR2 and CCR5). Also, iNOS expression remained low, while ARG1 was upregulated, indicating predominance of a M2-type response. At later points in time, the bone matrix density and volume were increased, in agreement with a high expression of Col1a1 and Col21a2. The remodelling process was marked by peaks of MMPs, RANKL and OPG expression at 14 days, and an increased density of osteoclasts. At 21 days, intimate Ti/bone contact was observed, with expression of final osteoblast differentiation markers (PHEX, SOST), as well as red spectrum collagen fibers. Conclusions: This study demonstrated a unique molecular view of oral osseointegration kinetics in C57Bl/6 mice, evidencing potential elements responsible for orchestrating cell migration, proliferation, ECM deposition and maturation, angiogenesis, bone formation and remodeling at the bone-implant interface in parallel with a novel microscopic analysis.

Efectos divergentes del factor de crecimiento endotelial vascular, VEGF y el fragmento N-terminal de la proteína relacionada con la parathormona, PTHrP en células madre mesenquimales derivadas de tejido adiposo humano / No disponible

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El factor de crecimiento endotelial vascular (VEGF) y el fragmento N-terminal de la proteína relacionada con la parathormona (PTHrP) regulan la proliferación de células madre mesenquimales humanas / Vascular endothelial growth factor (VEGF) and the N-terminal portion of parathyroid hormone-related protein (PTHrP) regulate the proliferation of human mesenchymal stem cells

El tejido adiposo contiene un gran número de células madre mesenquimales (Adipose Stem Cells, ASCs) que residen en su estroma vascular. Aunque existe controversia acerca de la capacidad de generar tejido óseo de estas células in vivo, in vitro constituyen un buen modelo de diferenciación osteogénica debido a su semejanza fenotípica con las células estromales de la médula ósea (Bone Marrow Stromal Cells, BMSCs) en cultivo. La diferenciación de las poblaciones osteoprogenitoras de la médula ósea está intensamente regulada por factores locales, como el factor de crecimiento endotelial vascular (VEGF) y la proteína relacionada con la parathormona (PTHrP), que modulan la proliferación de estas poblaciones en distintos estadios de diferenciación. Tanto el VEGF como el fragmento N-terminal de la PTHrP ejercen efectos osteogénicos. En este estudio hipotetizamos que sus efectos sobre la proliferación celular de los osteoprogenitores son dependientes del estadio de diferenciación osteoblástica. Tras confirmar su capacidad de diferenciación in vitro por expresión génica de Runx2 y acumulación de calcio, se analizó la respuesta proliferativa a estímulos con VEGF o PTHrP(1-36) de ASCs sometidas o no a inducción osteogénica. VEGF pero no PTHrP(1-36) estimuló la capacidad proliferativa de las ASCs no inducidas mientras que PTHrP(1-36), pero no VEGF, estimuló la proliferación de las ASCs inducidas, corroborando el papel diferencial de estos factores de crecimiento en distintos estadios de diferenciación (AU)

Adipose tissue contains a large number of mesenchymal stem cells (ASCs) residing in their vascular stroma. Although there is controversy regarding the ability to generate bone tissue from these cells in vivo, the in vitro cells offer a good model of osteogenic differentiation due to its phenotypic similarity with the bone marrow stromal cells (BMSCs) in culture. The differentiation of osteo-progenitor populations of bone marrow is intensely regulated by local factors, such as vascular endothelial growth factor (VEGF) and parathyroid hormone-related protein (PTHrP), which modulate these populations' proliferation in different stages of differentiation. Both the VEGF and the N-terminal fragment of the PTHrP exert osteogenic effects. In this study, we posited that its effects on proliferation of osteo-progenitors are stage dependent of osteoblastic differentiation. After confirming its capacity to in vitro differentiation by Runx2 gene expression and accumulation of calcium, the proliferative response to stimuli was analyzed with VEGF or PTHrP (1-36) of ASCs submitted or not to osteogenic induction. VEGF, but not PTHrP (1- 36), stimulated the proliferative capacity of uninduced ASCs, whereas BMSCs, but not VEGF, stimulated the proliferation of induced ASCs, corroborating the differential role of this growth in different stages of differentiation (AU)

[Role of immunological factors in the development of myopic choroidal neovascularization]. / Vliyanie immunologicheskikh faktorov na mekhanizmy formirovaniya miopicheskoi khorioidal'noi neovaskulyarizatsii.

AIM: To study the concentration of cytokines in the aqueous humor of the anterior chamber in patients with myopic choroidal neovascularization (mCNV) and to compare the results to their ophthalmic status. MATERIAL AND METHODS: A total of 19 patients (19 eyes) with mCNV treated with intravitreal ranibizumab were included in the study. The control group consisted of 15 patients (15 eyes) with myopia who had cataract surgery. Age, sex, and refractive error distribution were similar to that in the study group. All patients underwent a detailed ophthalmic examination as well as immunological study of the aqueous humor for cytokines concentrations using flow fluorometry (Bio-Plex Pro Human Cytokine Panel, 27-Plex, Bio-Rad Laboratories, USA). RESULTS: Significant differences in concentrations of 10 cytokines were found between the mCNV and study groups. Vascular endothelial growth factor (VEGF) level was twice as low in patients with mCNV as that in the controls (191.15±142.3 pg/ml and 320.06±170.05 pg/ml, respectively) (p<0.05). The other 9 cytokines were higher in mCNV, namely, platelet-derived growth factor (PDGF-BB), inflammatory and anti-inflammatory cytokines (IL-2, IL-15, IL-17А and IL-5, IL-13, respectively), tumor necrosis factor (TNFα), and chemokines (IL-8, RANTES). The degree of myopia as well as morphological and functional changes in the macular zone were shown to be in close correlation with cytokines involved in inflammation and VEGF. VEGF level appeared to be negatively related to axial eye length, refractive error, and three cytokines: IL-13, INF-γ, and RANTES. At the same time, numerous (6, 8 and more) close correlations were established between inflammatory and anti-inflammatory cytokines, chemokines, and growth factors. CONCLUSION: Patients with mCNV have been found to have higher than usual levels of inflammatory and anti-inflammatory cytokines, chemokines, and growth factors as well as a significantly decreased VEGF concentration. Immunological status of these patients differs from that in other ocular neovascular diseases suggesting possible involvement of alternative pathogenetic mechanisms.

Detection of angiogenic factors in midtrimester amniotic fluid and the prediction of preterm birth.

OBJECTIVE: We investigated whether the level of vascular endothelial growth factor (VEGF), placental growth factor (PlGF), and soluble VEGF receptor-1 (sFlt-1) in midtrimester amniotic fluid of preterm birth have different values compared with term delivery. MATERIALS AND METHODS: Our participants were 86 pregnant women who had undergone amniocentesis from 16 to 19 weeks of gestation. Forty-three cases were women with preterm delivery, and the other 43 cases were matched women with full-term delivery. Stored amniotic fluid was investigated after the delivery. The levels of VEGF, PlGF, and sFlt-1 were measured by enzyme-linked immunosorbent assay and Western blot. RESULTS: The levels of VEGF and PlGF in the preterm group were significantly higher than in the control group (30.48 ± 8.57 pg/mL vs. 26.06 ± 8.24 pg/mL and 28.83 ± 7.83 pg/mL vs. 25.35 ± 8.26 pg/mL, respectively) (p = 0.017 and 0.048, respectively). In terms of sFlt-1, the levels were decreased in the preterm group (10,478.51 ± 4012.56 pg/mL vs. 12,544.05 ± 4140.96 pg/mL) (p = 0.021). CONCLUSION: This study explains that elevated levels of VEGF and PlGF, suggestive of angiogenesis and tendency of inflammation at midtrimester, are predictive of preterm delivery, and their availability is maximized by downregulation of sFlt-1.

Mean platelet volume and platelet distribution as a diagnostic marker for penile vascular disease in patients with erectile dysfunction / Volumen medio de plaquetas y distribución de plaquetas como marcador diagnóstico para la enfermedad vascular peneana en pacientes con disfunción eréctil

Objective. To investigate the relationship between platelet activation parameters that are involved in the vascular response, the atherothrombotic process, and erectile function, in which epithelial dysfunction plays a significant role. Materials and methods. A study was performed on patients who had a color Doppler ultrasound (CDUS) of the penis due to erectile dysfunction. The patients were divided into two groups: those with normal CDUS findings, and those with vascular dysfunction on CDUS. Patients were also divided into two groups according to their scores using the International Index of Erectile Function (IIEF). The relationships between platelet activation factors, vascular parameters, and severity of the disorder were analyzed. Results. A total of 91 patients who fulfilled the inclusion criteria were included in the study. CDUS showed vascular dysfunction in 55 patients (Group I), while the findings were normal in 36 patients (Group II). Age, cholesterol level, disease duration, mean platelet volume (MPV), and platelet distribution width (PDW) were compared between Groups I and II, with Group I showing significantly higher values. The parameters that could affect Doppler ultrasound results were analyzed using multivariate regression analysis. This showed that PDW and disease duration were independent prognostic factors (p = .021 and p = .005, respectively). When the patients were divided into two groups according to their IIEF scores, in those with mild (Group A) and severe disease (Group B), it was found that there were significant differences between the groups with age, disease duration, and PDW, while two groups were found similar in terms of MPV, cholesterol levels, and hormone parameters. Conclusion. It is supposed that increased platelet activation parameters, and PDW in particular, give important information for disease progression and follow-up of vascular dysfunction in erectile dysfunction (AU)

Objetivo. Investigar la relación entre los parámetros de activación de plaquetas involucrados en la respuesta vascular, el proceso aterotrombótico y la función eréctil, en la que la disfunción endotelial desempeña un papel importante. Material y métodos. Se analizaron los pacientes en los que se realizó una ecografía Doppler color (EDC) del pene por disfunción eréctil. Los pacientes fueron divididos en 2 grupos: con resultados normales en la EDC y con disfunción vascular en la EDC. Los pacientes fueron asimismo divididos en 2 grupos en función de las puntuaciones del Índice Internacional de Función Eréctil (IIFE). Se analizaron las relaciones entre los factores de activación plaquetaria, los parámetros vasculares y la gravedad del trastorno. Resultados. Un total de 91 pacientes que cumplieron los criterios de inclusión fueron incluidos en el estudio. La EDC mostró disfunción vascular en 55 pacientes (grupo i), mientras que los resultados fueron normales en 36 pacientes (grupo ii). La edad, el nivel de colesterol, la duración de la enfermedad, el volumen medio de plaquetas (VMP) y la anchura de distribución de las plaquetas (ADP) se compararon entre los grupos i y ii; además, los valores fueron expresivamente más altos en el grupo i. Los parámetros que podrían afectar los resultados de la EDC se valoraron con el análisis de regresión multivariante, lo que demostró que la ADP y la duración de la enfermedad fueron factores pronósticos independientes (p = 0,021 y p = 0,005, respectivamente). Cuando los pacientes fueron divididos en 2 grupos en función de las puntuaciones IIFE, en aquellos con enfermedad leve (grupo A) y enfermedad grave (grupo B) se encontraron diferencias significativas entre los grupos con la edad, la duración de la enfermedad y la ADP, mientras que ambos grupos fueron similares en términos de VMP, de niveles de colesterol y de parámetros hormonales. Conclusión. Al parecer, el aumento de los parámetros de activación plaquetar, y especialmente la ADP, proporcionan una importante información para la progresión de la enfermedad y el seguimiento de la disfunción vascular en la disfunción eréctil (AU)