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1.
Biochem Cell Biol ; 102(1): 47-59, 2024 Feb 01.
Article in English | MEDLINE | ID: mdl-37459649

ABSTRACT

Hypoxia, low, non-physiological oxygen tension is a key regulator of tumor microenvironment, determining the pathological tumor vascularization. Alleviation of hypoxia through vessel normalization may be a promising therapeutic approach. We aimed to assess the role of low oxygen tension in PTEN-related pathways and proangiogenic response, in vitro, in two different tumor cell lines, focusing on potential therapeutic targets for tumor vessel normalization. Downregulation of PTEN in hypoxia mediates the activation of distinct mechanisms: cytoplasmic pAKT activation in melanoma and pMDM2 modulation in kidney cancer. We show that hypoxia-induced proangiogenic potential was stronger in Renca cells than B16 F10-confirmed by a distinct secretory potential and different ability to affect endothelial cells functions. Therefore, the impact of hypoxia on PTEN-mediated regulation may determine the therapeutic targets and effectiveness of vessel normalization and intrinsic characteristics of cancer cell have to be taken into account when designing treatment.


Subject(s)
Endothelial Cells , Melanoma , Humans , Endothelial Cells/metabolism , Hypoxia , Cell Line, Tumor , Neovascularization, Pathologic/pathology , Oxygen , Tumor Microenvironment , PTEN Phosphohydrolase/metabolism
2.
Int J Mol Sci ; 24(6)2023 Mar 16.
Article in English | MEDLINE | ID: mdl-36982788

ABSTRACT

Natural compounds, such as resveratrol (Res), are currently used as adjuvants for anticancer therapies. To evaluate the effectiveness of Res for the treatment of ovarian cancer (OC), we screened the response of various OC cell lines to the combined treatment with cisplatin (CisPt) and Res. We identified A2780 cells as the most synergistically responding, thus optimal for further analysis. Because hypoxia is the hallmark of the solid tumor microenvironment, we compared the effects of Res alone and in combination with CisPt in hypoxia (pO2 = 1%) vs. normoxia (pO2 = 19%). Hypoxia caused an increase (43.2 vs. 5.0%) in apoptosis and necrosis (14.2 vs. 2.5%), reactive oxygen species production, pro-angiogenic HIF-1α (hypoxia-inducible factor-1α) and VEGF (vascular endothelial growth factor), cell migration, and downregulated the expression of ZO1 (zonula occludens-1) protein in comparison to normoxia. Res was not cytotoxic under hypoxia in contrast to normoxia. In normoxia, Res alone or CisPt+Res caused apoptosis via caspase-3 cleavage and BAX, while in hypoxia, it reduced the accumulation of A2780 cells in the G2/M phase. CisPt+Res increased levels of vimentin under normoxia and upregulated SNAI1 expression under hypoxia. Thus, various effects of Res or CisPt+Res on A2780 cells observed in normoxia are eliminated or diminished in hypoxia. These findings indicate the limitations in using Res as an adjuvant with CisPt therapy in OC.


Subject(s)
Cisplatin , Ovarian Neoplasms , Humans , Female , Cisplatin/pharmacology , Cisplatin/therapeutic use , Ovarian Neoplasms/metabolism , Resveratrol/pharmacology , Vascular Endothelial Growth Factor A/metabolism , Cell Line, Tumor , Hypoxia , Vascular Endothelial Growth Factors/metabolism , Cell Hypoxia , Hypoxia-Inducible Factor 1, alpha Subunit/metabolism , Tumor Microenvironment
3.
J Cell Mol Med ; 25(7): 3284-3299, 2021 04.
Article in English | MEDLINE | ID: mdl-33624446

ABSTRACT

Pathologic angiogenesis directly responds to tumour hypoxia and controls the molecular/cellular composition of the tumour microenvironment, increasing both immune tolerance and stromal cooperation with tumour growth. Myo-inositol-trispyrophosphate (ITPP) provides a means to achieve stable normalization of angiogenesis. ITPP increases intratumour oxygen tension (pO2 ) and stabilizes vessel normalization through activation of endothelial Phosphatase-and-Tensin-homologue (PTEN). Here, we show that the tumour reduction due to the ITPP-induced modification of the tumour microenvironment by elevating pO2 affects the phenotype and properties of the immune infiltrate. Our main observations are as follows: a relative change in the M1 and M2 macrophage-type proportions, increased proportions of NK and CD8+ T cells, and a reduction in Tregs and Th2 cells. We also found, in vivo and in vitro, that the impaired access of PD1+ NK cells to tumour cells is due to their adhesion to PD-L1+ /PD-L2+ endothelial cells in hypoxia. ITPP treatment strongly reduced PD-L1/PD-L2 expression on CD45+/CD31+ cells, and PD1+ cells were more numerous in the tumour mass. CTLA-4+ cell numbers were stable, but level of expression decreased. Similarly, CD47+ cells and expression were reduced. Consequently, angiogenesis normalization induced by ITPP is the mean to revert immunosuppression into an antitumor immune response. This brings a key adjuvant effect to improve the efficacy of chemo/radio/immunotherapeutic strategies for cancer treatment.


Subject(s)
Antineoplastic Agents/pharmacology , Cell Hypoxia , Inositol Phosphates/pharmacology , Neovascularization, Pathologic/drug therapy , Tumor Microenvironment , Animals , Antineoplastic Agents/therapeutic use , B7-H1 Antigen/metabolism , Cell Line, Tumor , Inositol Phosphates/therapeutic use , Lymphocytes/drug effects , Lymphocytes/immunology , Macrophages/drug effects , Macrophages/immunology , Mice , Mice, Inbred BALB C , Mice, Inbred C57BL , Mice, Nude , Neovascularization, Pathologic/immunology , PTEN Phosphohydrolase/metabolism , Programmed Cell Death 1 Ligand 2 Protein/metabolism , Tumor Cells, Cultured
4.
Int J Mol Sci ; 22(2)2021 Jan 07.
Article in English | MEDLINE | ID: mdl-33430201

ABSTRACT

Endothelial cells (ECs) lining the blood vessels are important players in many biological phenomena but are crucial in hypoxia-dependent diseases where their deregulation contributes to pathology. On the other hand, processes mediated by ECs, such as angiogenesis, vessel permeability, interactions with cells and factors circulating in the blood, maintain homeostasis of the organism. Understanding the diversity and heterogeneity of ECs in different tissues and during various biological processes is crucial in biomedical research to properly develop our knowledge on many diseases, including cancer. Here, we review the most important aspects related to ECs' heterogeneity and list the available in vitro tools to study different angiogenesis-related pathologies. We focus on the relationship between functions of ECs and their organo-specificity but also point to how the microenvironment, mainly hypoxia, shapes their activity. We believe that taking into account the specific features of ECs that are relevant to the object of the study (organ or disease state), especially in a simplified in vitro setting, is important to truly depict the biology of endothelium and its consequences. This is possible in many instances with the use of proper in vitro tools as alternative methods to animal testing.


Subject(s)
Cell Hypoxia/genetics , Cellular Microenvironment/genetics , Endothelial Cells/cytology , Neovascularization, Physiologic/genetics , Blood Vessels/growth & development , Blood Vessels/metabolism , Human Umbilical Vein Endothelial Cells , Humans , Induced Pluripotent Stem Cells/cytology
5.
Pol J Radiol ; 86: e136-e142, 2021.
Article in English | MEDLINE | ID: mdl-33758639

ABSTRACT

PURPOSE: Transarterial chemoembolization (TACE) is currently recommended for unresectable intrahepatic tumours with no vascular invasion or metastasis to other organs. It is based on drug-eluting microspheres pre-loaded with chemotherapeutics, which are injected selectively into vessels supplying the tumour, to embolize them inducing ischaemia, and elute the drug, to induce tumour response. We present our initial experience with novel irinotecan- loaded Embocure Plus microspheres in patients with metastatic colorectal cancer tumours in the liver, and their effect on HCT-116 cell cultures in vitro. MATERIAL AND METHODS: Three consecutive male patients (median age 62 [50-76] years) with liver metastatic colorectal cancer tumours were selected. All patients had a pre-procedure contrast-enhanced computed tomography, confirming multiple metastatic liver tumours (mean tumour diameter = 42 mm; range: 14-77 mm) and periprocedural dyna-CT scans for rapid treatment results assessment. In vitro: Human colon HCT116 cancer cell line was cultured, irinotecan loaded Embocure Plus microspheres were added. Cultures were assessed after 24 hours and 72 hours of incubation in normoxia or hypoxia. RESULTS: All embolizations were technically successful, and no complications were observed. Stabilization of the targeted metastatic liver tumours in all patients was noted. In vitro: Significant decrease of the growth of HTC 116 cell lines were observed in controls compared to cells treated with Embocure Plus loaded with irinotecan in normoxia and hypoxia after 48 and 72 hours. We observed a tendency for less inhibited cell proliferation in low-oxygen conditions. CONCLUSIONS: TACE therapy of liver metastatic tumours shows satisfactory results and a low complication rate. Embocure Plus microspheres are safe and technically feasible for superselective chemoembolization of metastatic colorectal cancer liver tumour. Dyna-CT can be used for assessment of treatment results during repeated TACE procedures.

6.
J Sex Med ; 14(1): 125-133, 2017 01.
Article in English | MEDLINE | ID: mdl-27989490

ABSTRACT

INTRODUCTION: Rape and pedophilic child molestation are the most commonly convicted sexual offenses in Poland. Recent studies have suggested a possible genetic contribution toward pathologic sexual interests and behaviors. AIM: To analyze and compare functional polymorphisms of genes associated with the activity of the serotonin and dopamine systems in a group of paraphilic sexual offenders and control subjects. METHODS: The study sample (n = 97) consisted of two groups: paraphilic sexual offenders (65 pedophilic child molesters and 32 rapists) and controls (n = 76). Genetic polymorphisms previously associated with behavioral control, addictive behaviors, and sexual functions were chosen for analyses. Specifically, functional polymorphisms in dopamine receptors genes (DRD1, DRD2, DRD4), catechol-O-methyltransferase gene (COMT), dopamine transporter gene (DAT), serotonin transporter gene (SLC6A4), serotonin type 2A receptor gene (5HTR2A), tryptophan hydroxylase 2 gene (TPH2), monoamine oxidase A gene (MAOA), and brain-derived neurotrophic factor gene (BDNF) were analyzed. MAIN OUTCOME MEASURES: An association between a history of sexual offense and the distribution of genotypes and alleles in the analyzed polymorphisms. RESULTS: Our results found no association between a history of sexual offense and the distribution of genotypes or alleles in the analyzed polymorphisms. CONCLUSION: Although these results are limited by the small sample and are exploratory, they highlight a novel approach to sample selection in a population that is difficult to access and study. Future research should include larger samples and other relevant polymorphisms to advance this field of study.


Subject(s)
Criminals , Dopamine/metabolism , Pedophilia/genetics , Serotonin/metabolism , Adult , Alleles , Case-Control Studies , Catechol O-Methyltransferase/genetics , Child , Genotype , Humans , Male , Middle Aged , Poland , Polymorphism, Genetic , Sex Offenses
7.
Pharmacol Rep ; 75(5): 1187-1199, 2023 Oct.
Article in English | MEDLINE | ID: mdl-37673853

ABSTRACT

BACKGROUND: PTEN is a tumor suppressor that is often mutated and nonfunctional in many types of cancer. The high heterogeneity of PTEN function between tumor types makes new Pten knockout models necessary to assess its impact on cancer progression and/or treatment outcomes. METHODS: We aimed to show the effect of CRISPR/Cas9-mediated Pten knockout on murine melanoma (B16 F10) and kidney cancer (Renca) cells. We evaluated the effect of PTEN deregulation on tumor progression in vivo and in vitro, as well as on the effectiveness of drug treatment in vitro. In addition, we studied the molecular changes induced by Pten knockout. RESULTS: In both models, Pten mutation did not cause significant changes in cell proliferation in vitro or in vivo. Cells with Pten knockout differed in sensitivity to cisplatin treatment: in B16 F10 cells, the lack of PTEN induced sensitivity and, in Renca cells, resistance to drug treatment. Accumulation of pAKT was observed in both cell lines, but only Renca cells showed upregulation of the p53 level after Pten knockout. PTEN deregulation also varied in the way that it altered PAI-1 secretion in the tested models, showing a decrease in PAI-1 in B16 F10 Pten/KO and an increase in Renca Pten/KO cells. In kidney cancer cells, Pten knockout caused changes in epithelial to mesenchymal transition marker expression, with downregulation of E-cadherin and upregulation of Snail, Mmp9, and Acta2 (α-SMA). CONCLUSIONS: The results confirmed heterogenous cell responses to PTEN loss, which may lead to a better understanding of the role of PTEN in particular types of tumors and points to PTEN as a therapeutic target for personalized medicine.


Subject(s)
Carcinoma, Renal Cell , Kidney Neoplasms , Melanoma , PTEN Phosphohydrolase , Animals , Mice , Cell Line, Tumor , Drug Resistance , Epithelial-Mesenchymal Transition , Kidney Neoplasms/drug therapy , Kidney Neoplasms/genetics , Melanoma/drug therapy , Melanoma/genetics , Plasminogen Activator Inhibitor 1 , PTEN Phosphohydrolase/genetics
8.
Front Cell Dev Biol ; 11: 1125077, 2023.
Article in English | MEDLINE | ID: mdl-37261072

ABSTRACT

Introduction: Hypoxia shapes the tumor microenvironment, modulates distinct cell population activities, and activates pathological angiogenesis in cancer, where endothelial cells (ECs) are the most important players. This study aimed to evidence the influences of the tumor microenvironment on the global gene expression pattern characteristic for ECs and the distinct responses displayed by tumor-derived ECs in comparison to the healthy endothelium during endothelial to mesenchymal transition (EndMT) and its regulation by miR-200-b-3p. Methodology: Immortalized lines of ECs from the same patient with breast cancer, healthy breast tissue (HBH.MEC), and primary tumor (HBCa.MEC) were used. The experiments were performed in normoxia and hypoxia for 48 h. By using the wound healing test, we investigated the migration abilities of ECs. Global gene expression analysis with NGS was carried out to detect new pathways altered in pathological ECs and find the most changed miRNAs. The validation of NGS data from RNA and miRNA was estimated by qPCRs. Mimic miR-200b-3p was used in HBH.MEC, and the targets VEGF, Bcl2, ROCK2, and SP1 were checked. Results: Hypoxia influences EC migration properties in wound healing assays. In hypoxia, healthy ECs migrate slower than they do in normoxia, as opposed to HBCa.MEC, where no decreased migration ability is induced by hypoxia due to EndMT features. NGS data identified this process to be altered in cancer ECs through extracellular matrix (ECM) organization. The deregulated genes, validated by qPCR, included SPP1, ITGB6, COL4A4, ADAMST2, LAMA1, GAS6, PECAM1, ELN, FBLN2, COL6A3, and COL9A3. NGS also identified collagens, laminins, fibronectins, and integrins, as being deregulated in tumor-derived ECs. Moreover, the analysis of the 10 most intensively modified miRNAs, when breast tumor-derived ECs were compared to healthy ECs, shed light on miR-200b-3p, which is strongly upregulated in HBCa.MECs when compared to HBH.MECs. Discussion and conclusion: The pathological ECs differed significantly, both phenotypically and functionally, from the normal corresponding tissue, thus influencing their microenvironment cross-talk. The gene expression profile confirms the EndMT phenotype of tumor-derived ECs and migratory properties acquisition. Moreover, it indicates the role of miR-200b-3p, that is, regulating EndMT in pathological ECs and silencing several angiogenic growth factors and their receptors by directly targeting their mRNA transcripts.

9.
Bioelectrochemistry ; 150: 108363, 2023 Apr.
Article in English | MEDLINE | ID: mdl-36608369

ABSTRACT

The abnormal concentration of microRNAs (miRNAs) can be associated with occurrence of various diseases including cancer, cardiovascular and neurodegenerative, hence they can be considered as potential biomarkers. An attractive approach could be the application of electrochemical methods, particularly where hybridization event between single-stranded deoxyribonucleic acid (ssDNA) or peptide-nucleic acid (PNA) with miRNA strand happens. Recently, the use of various nanomaterials such as gold nanoparticles, graphene oxide, quantum dots as well as catalyzed hairpin assembly or hybridization chain reaction were proposed to further enhance the performance of elaborated sensors. Herein, we present the studies on selection of receptor layer composition for detection of miRNA 141. The possibility of formation of receptor layer and further duplex monolayer between ssDNA or PNA with miRNA was analyzed by atomic force microscopy (AFM) technique. The interaction of ssDNA and PNA probes with miRNA was further verified using surface plasmon resonance (SPR) and quartz - crystal microbalance (QCM) techniques. On the basis of impedance spectroscopy it was shown that the use of unlabelled ssDNA as receptor layer provided 0.1 pM detection limit. This shows that proposed biosensor that is simple in preparation and use is an attractive alternative to other recently presented approaches.


Subject(s)
Biosensing Techniques , Metal Nanoparticles , MicroRNAs , Peptide Nucleic Acids , Peptide Nucleic Acids/chemistry , DNA, Single-Stranded , Gold/chemistry , Biosensing Techniques/methods , Nucleic Acid Hybridization , Limit of Detection , DNA Probes
10.
In Vitro Cell Dev Biol Anim ; 58(5): 349-364, 2022 May.
Article in English | MEDLINE | ID: mdl-35536385

ABSTRACT

Tumor development studies should adapt to cancer cells' specific mechanisms in connection with their microenvironment. Standard two-dimensional cultures and gas composition are not relevant to the real cancer environment. Existing three-dimensional models are often requiring sophisticated conditions. Here, we propose and characterize, in two cancer models, melanoma (B16F10) and kidney cancer (RenCa), a three-dimensional culture method, reporting the presence of hypoxia-related genes/proteins and aggressiveness mechanisms (epithelial mesenchymal transition and cancer stem cells). We validate the designed three-dimensional method by comparing it with in vivo growing tumors. The developed method brings simplicity and data reproducibility. Melanoma spheroid-growing cells reached a cell cycle arrest at the G0/G1 phase and showed induction of hypoxia. Spheroid-recovered RenCa cells were enriched in proliferating cells and displayed delayed hypoxia. Moreover, the responses to hypoxia observed in spheroids were validated by in vivo tumor studies for both lines. Three-dimensional shapes induced cancer stem cells in renal cancer, whereas epithelial to mesenchymal transition occurred in the melanoma model. Such distinction in the use of different aggressiveness-leading pathways was observed in in vivo melanoma vs kidney tumors. Thus, this 3D culture model approach is adequate to uncover crucial molecular pathways using distinct mechanisms to reach aggressiveness; i.e., B16F10 cells perform epithelial to mesenchymal transition while RenCa cells dedifferentiate into cancer stem cells. Such three-dimensional models help mimic the in vivo tumor features, i.e., hypoxia and aggressiveness mechanisms as validated here by next-generation sequencing analysis, and are proposed for further alternative methods to in vivo studies.


Subject(s)
Kidney Neoplasms , Melanoma , Animals , Cell Line, Tumor , Epithelial-Mesenchymal Transition/genetics , Hypoxia , Kidney Neoplasms/genetics , Kidney Neoplasms/pathology , Melanoma/genetics , Melanoma/pathology , Reproducibility of Results , Spheroids, Cellular/pathology , Tumor Microenvironment
11.
Biomolecules ; 12(5)2022 05 11.
Article in English | MEDLINE | ID: mdl-35625614

ABSTRACT

MicroRNAs are post-transcriptional regulators of gene expression, and disturbances of their expression are the basis of many pathological states, including cancers. The miRNA pattern in the context of tumor microenvironment explains mechanisms related to cancer progression and provides a potential target of modern therapies. Here we show the miRNA pattern in renal cancer focusing on hypoxia as a characteristic feature of the tumor microenvironment and dysregulation of PTEN, being a major tumor suppressor. Methods comprised the CRSPR/Cas9 mediated PTEN knockout in the Renca kidney cancer cell line and global miRNA expression analysis in both in vivo and in vitro (in normoxic and hypoxic conditions). The results were validated on human cancer models with distinct PTEN status. The increase in miR-210-3p in hypoxia was universal; however, the hypoxia-induced decrease in PTEN was associated with an increase in miR-221-3p, the loss of PTEN affected the response to hypoxia differently by decreasing miR-10b-5p and increasing miR-206-3p. In turn, the complete loss of PTEN induces miR-155-5p, miR-100-5p. Upregulation of miR-342-3p in knockout PTEN occurred in the context of the whole tumor microenvironment. Thus, effective identification of miRNA patterns in cancers must consider the specificity of the tumor microenvironment together with the mutations of key suppressors.


Subject(s)
Kidney Neoplasms , MicroRNAs , Humans , Hypoxia/metabolism , Kidney Neoplasms/genetics , MicroRNAs/genetics , MicroRNAs/metabolism , PTEN Phosphohydrolase/genetics , PTEN Phosphohydrolase/metabolism , Transcriptional Activation , Tumor Microenvironment/genetics , Up-Regulation
12.
J Clin Med ; 12(1)2022 Dec 20.
Article in English | MEDLINE | ID: mdl-36614808

ABSTRACT

Mortality in COVID-19 is mainly associated with respiratory failure, cytokine storm, and macrophage activation. Oxygenation and anti-inflammatory effects of Hyperbaric Oxygen Therapy (HBOT) suggest that it is a promising adjunct treatment for COVID-19. Repeated sessions of HBO with standard COVID-19 therapy were used to reduce the inflammation and increase oxygenation. We evaluated the safety and efficacy of HBOT in avoiding the replacement ventilation and/or ECMO and its effect on the inflammatory process. Twenty-eight moderate-to-severe COVID-19 patients were randomized into control or HBOT group. HBOT patients participated in 5 hyperbaric sessions (60 min). Before and after each session blood gas levels and vital parameters were monitored. Blood samples were collected for extended biochemical tests, blood morphology and immunological assays. There were 3 deaths in the control, no deaths in the HBOT group. No adverse events leading to discontinuation of HBOT were observed and patients receiving HBOT required lower oxygen delivery. We observed decrease in CRP, ferritin and LDH and increase in CD3 in HBOT group compared to control. This study confirmed the feasibility and safety of HBOT in patients with COVID-19 and indicated HBOT can lead to alleviation of inflammation and partial restoration of T cell responses.

13.
J Clin Med ; 11(19)2022 Sep 23.
Article in English | MEDLINE | ID: mdl-36233473

ABSTRACT

During late pregnancy, sleep deterioration is regularly observed. In concert with these observations, in previous studies by other researchers, a slight objective cognitive decline in pregnant women has been found. Sleep is essential for memory consolidation. The hypothesis of the study was that cognitive impairment could be related to sleep deterioration during pregnancy. The study included 19 pregnant women in their third trimester of pregnancy (28−40 weeks, median 33 weeks (IQR 32−37)) recruited at the Department of Gynecology and Obstetrics, Medical University of Warsaw, and 20 non-pregnant women as controls. The assessment was performed using the vocabulary subtest from the Wechsler Adult Intelligence Scale (WAIS), D2 Test of Attention, OSPAN task (Operational Span Task) to assess cognitive performance, actigraphy to examine sleep parameters, and a set of self-report instruments: Athens Insomnia Scale (AIS), Beck Depression Inventory (BDI), Ford Insomnia Response to Stress (FIRST), Regenstein Hyperarousal Scale (HS), and Epworth Sleepiness Scale (ESS). Although there were no differences between the groups in WAIS (p = 0.18), pregnant women had worse scores in working memory capacity (overall number of remembered letters: p = 0.012, WM span index: p = 0.004) and a significantly lower score in attention (p = 0.03). Pregnant women also had lower sleep efficiency (p = 0.001), more awakenings from sleep (p = 0.001), longer average awakenings (p < 0.0001), longer wake after sleep onset (WASO, p < 0.0001), and longer total time in bed (p < 0.0001). In psychological assessment, pregnant women had only a higher FIRST score (p = 0.02). Using mediation analysis, we found that frequent awakening might be the major factor contributing to deterioration in working memory performance, explaining almost 40% of the total effect. In conclusion, sleep fragmentation in the third trimester of pregnancy may impair working memory consolidation. Pregnant women often complain about poor daily performance as well as non-restorative sleep. In this study, we showed that there is a relationship between lower sleep quality in pregnancy and worse cognitive functioning. We can expect a cognitive decline in women with sleep disturbances in pregnancy. Therefore, we should pay more attention to the treatment of sleep disorders in pregnancy.

14.
Pharmaceutics ; 14(7)2022 Jul 06.
Article in English | MEDLINE | ID: mdl-35890313

ABSTRACT

The failures of anti-ß-amyloid immunotherapies suggested that the very low fraction of injected antibodies reaching the brain parenchyma due to the filtering effect of the BBB may be a reason for the lack of therapeutic effect. However, there is no treatment, as yet, for the amyotrophic lateral sclerosis (ALS) despite substantial evidence existing of the involvement of TDP-43 protein in the evolution of ALS. To circumvent this filtering effect, we have developed a novel approach to facilitate the penetration of antibody fragments (Fabs) into the brain parenchyma. Leveraging the homing properties of endothelial progenitor cells (EPCs), we transfected, ex vivo, such cells with vectors encoding anti-ß-amyloid and anti-TDP43 Fabs turning them into an "antibody fragment factory". When injected these cells integrate into the BBB, where they secrete anti-TDP43 Fabs. The results showed the formation of tight junctions between the injected engineered EPCs and the unlabeled resident endothelial cells. When the EPCs were further modified to express the anti-TDP43 Fab, we could observe integration of these cells into the vasculature and the secretion of Fabs. Results confirm that production and secretion of Fabs at the BBB level leads to their migration to the brain parenchyma where they might exert a therapeutic effect.

15.
Sci Rep ; 11(1): 4472, 2021 02 24.
Article in English | MEDLINE | ID: mdl-33627727

ABSTRACT

We propose a new in vitro model to assess the impact of 90Y-microspheres derived low-dose beta radiation on colorectal cancer cell line under various oxygenation conditions that mimic the tumor environment. Cancer cells (HCT116) proliferation was assessed using Alamar Blue (AB) assay after 48, 72, and 96 h. FLUKA code assessed changes in cancer cell populations relative to the absorbed dose. In normoxia, mitochondrial activity measured by Alamar Blue after 48-72 h was significantly correlated with the number of microspheres (48 h: r = 0.87 and 72 h: r = 0.89, p < 0.05) and absorbed dose (48 h: r = 0.87 and 72 h: r = 0.7, p < 0.05). In hypoxia, the coefficients were r = 0.43 for both the number of spheres and absorbed dose and r = 0.45, r = 0.47, respectively. Impediment of cancer cell proliferation depended on the absorbed dose. Doses below 70 Gy could reduce colorectal cancer cell proliferation in vitro. Hypoxia induced a higher resistance to radiation than that observed under normoxic conditions. Hypoxia and radiation induced senescence in cultured cells. The new in vitro model is useful for the assessment of 90Y radioembolization effects at the micro-scale.


Subject(s)
Beta Particles/therapeutic use , Colorectal Neoplasms/radiotherapy , Yttrium Radioisotopes/administration & dosage , Cell Proliferation/radiation effects , HCT116 Cells , Humans , Hypoxia/radiotherapy , Microspheres , Mitochondria/radiation effects , Radiometry/methods
16.
Addict Behav ; 79: 8-13, 2018 04.
Article in English | MEDLINE | ID: mdl-29227790

ABSTRACT

INTRODUCTION: Prior work largely confirms the presence of various emotional processing deficits among individuals with an alcohol use disorder (AUD); however, their specificity and relevance still warrant investigation. The aim of the current study was to compare selected aspects of emotional processing (i.e., mental state recognition, alexithymia, and emotional intelligence) between individuals treated for an AUD and healthy individuals. METHODS: The AUD sample consisted of 92 abstinent men with AUD who were participating in an 8-week inpatient abstinence-based treatment program in Warsaw, Poland. The healthy control (HC) group consisted of 86 men recruited from the Medical University of Warsaw and the Nowowiejski Hospital administrative staff. Baseline information about demographics, psychopathological symptoms, and severity of alcohol problems was obtained. Mental states recognition was assessed using the Reading the Mind in the Eyes Test (RMET). Alexithymia was measured with the Toronto Alexithymia Scale (TAS-20). The Schutte Self-Report Emotional Intelligence Test (SSEIT) was used to measure emotional intelligence (EI). RESULTS AND CONCLUSIONS: After accounting for potentially confounding variables (demographics, severity of depression, anxiety symptoms) in MANCOVA models, patients with AUD presented deficits in identification and description of their own emotional states, as well as lower emotion regulation skills when compared to HCs. No between-group differences were observed in self-reported recognition of other people's emotions, social skills, and a behavioral measure of mental states recognition. Specific rather than general emotion-processing deficits in participants with AUD were identified, suggesting problems with processing of intrapersonal emotional signals.


Subject(s)
Alcoholism/psychology , Emotions , Interpersonal Relations , Theory of Mind , Adult , Affective Symptoms/psychology , Anxiety/psychology , Case-Control Studies , Depression/psychology , Emotional Intelligence , Humans , Male , Middle Aged , Young Adult
17.
Ann Acad Med Stetin ; 53(3): 148-56, 2007.
Article in Polish | MEDLINE | ID: mdl-18595498

ABSTRACT

INTRODUCTION: Cancer prophylaxis seems nowadays to be the more and more powerful tool in fight with these serious diseases. The aim of this work is to find out opinions of students of nursing extension studies on contemporary cancer prophylaxis. MATERIAL AND METHODS: The question about possibilities of practical efforts for prophylaxis and early detection of cancer was directed to 160 students of four consecutive years (2002-2006), at the end of the fourth year of lasting five and a half years extension nursing studies, during ending exam on subject: oncological nursing. There were 154 women and 6 men, predominantly at their third decade of life, with nursing experience approximately more than 5 years. RESULTS: Out of 160 asked students, 131 of them firstly indicated necessity of breast cancer prophylaxis, 117 mentioned lung cancer, 113 cervix cancer, 95 colorectal cancer, 33 prostate cancer. In families with cancer problems, more frequent control investigations (23 answers), and genetic tests (16) were called for. Patients should be qualified to appropriate risk groups (13) and controlled more frequently there (24). Apart from necessary wide education in media (126) personal contact with patient to discuss his or her personal problems relating to cancer is needed (91). If atypical symptoms are self-detected by patients it should alert them to not neglect and contact family physician (33). Healthy diet (62) containing fresh vegetables and fruits (73), high fibre diet (42) with less animal fat (38) and less red meat (30), containing no preservative agents (45) is recommended. Increased physical activity (84) to cease or reduce smoking (102), and alcohol intake (55), limited exposition to ultraviolet rays (49), and systematic controls of breast (105), uterus cervix (88), lungs (77), colon (55) and prostate (28) are proposed. The pollution of environment by combustion gases and smokes (34) not excluding risk factors of medical workplace (29) are mentioned as cancerogenic factors. CONCLUSIONS: In the time of increasing incidence of cancer in Poland it is necessary to elaborate a complex model of primary cancer prophylaxis and early detection of it and to implement it in practice. For better effectiveness of such systemic solution wide involvement of nursing staff is mandatory. Nurses should be, very well and consciously prepared to play their role through adequate educational efforts undertaken during nursing studies.


Subject(s)
Health Behavior , Health Knowledge, Attitudes, Practice , Life Style , Neoplasms/epidemiology , Neoplasms/prevention & control , Students, Nursing/statistics & numerical data , Adult , Alcohol Drinking/epidemiology , Alcohol Drinking/prevention & control , Breast Neoplasms/epidemiology , Breast Neoplasms/prevention & control , Colorectal Neoplasms/epidemiology , Colorectal Neoplasms/prevention & control , Comorbidity , Environmental Pollution/prevention & control , Exercise , Female , Humans , Incidence , Lung Neoplasms/epidemiology , Lung Neoplasms/prevention & control , Male , Poland/epidemiology , Population Surveillance , Prostatic Neoplasms/epidemiology , Prostatic Neoplasms/prevention & control , Smoking/epidemiology , Smoking Prevention , Uterine Cervical Neoplasms/epidemiology , Uterine Cervical Neoplasms/prevention & control
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