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Cell migration is a fundamental and complex phenomenon that occurs in normal physiology and in diseases like cancer. Hence, understanding cell migration is very important in the fields of developmental biology and biomedical sciences. Cell migration occurs in 3 dimensions (3D) and involves an interplay of migrating cell(s), neighboring cells, extracellular matrix, and signaling molecules. To understand this phenomenon, most of the currently available techniques still rely on 2-dimensional (2D) cell migration assay, also known as the scratch assay or the wound healing assay. These methods suffer from limited reproducibility in creating a cell-free region (a scratch or a wound). Mechanical/heat related stress to cells is another issue which hampers the applicability of these methods. To tackle these problems, we developed an alternative method based on 3D printed biocompatible cell inserts, for quantifying cell migration in 24-well plates. The inserts were successfully validated via a high throughput assay for following migration of lung cancer cell line (A549 cell line) in the presence of standard cell migration promoters and inhibitors. We also developed an accompanying image analysis pipeline which demonstrated that our method outperforms the state-of-the-art methodologies for assessing the cell migration in terms of reproducibility and simplicity.

Tématem této bakalářské práce jsou buněčné nosiče. V úvodu jsou definovány pojmy, jako regenerativní medicína, tkáňové inženýrství a její hlavní směry a cíle. Hlavní část práce je věnována klasifikaci buněčných nosičů, možnostem jejich využití, popisu jejich vlastností, charakterizaci materiálů využívaných k jejich výrobě a dále také popisu konkrétních způsobů výroby některých z nich. Pozornost je také věnována procesu decelularizace se zmíněnými možnostmi použití v rámci tkáňového inženýrství. V závěrečné části práce jsou shrnuty konkrétní příklady nosičů, které jsou nejčastěji využívány v klinické a experimentální medicíně a jsou uvedeny konkrétní příklady klinických studií., The topic of this bachelor thesis is cell carriers. In the introductory part, terms such as regenerative medicine, tissue engineering and its main directions and goals are defined. The main part of the thesis is devoted to the classification of cell carriers, their possible applications, description of their properties, characterization of the materials used for their production and also description of specific methods of production of some of them. Particular attention is paid to the process of decellularization with the mentioned possibilities of application in tissue engineering. In the final part of the thesis, specific examples of carriers that are most commonly used in clinical and experimental medicine are summarized and specific examples of clinical studies are given., Fakulta chemicko-technologická, 1. Prezentace výsledků bakalářské práce. 2. Diskuze k posudku vedoucího bakalářské práce. 3. Student/ka zodpověděl/a všechny dotazy a připomínky k bakalářské práci., Dokončená práce s úspěšnou obhajobou

Cerebral complications in preeclampsia contribute substantially to maternal mortality and morbidity. There is a lack of reliable and accessible predictors for preeclampsia-related cerebral complications. In this study, plasma from women with preeclampsia (n = 28), women with normal pregnancies (n = 28) and non-pregnant women (n = 16) was analyzed for concentrations of the cerebral biomarkers neurofilament light (NfL), tau, neuron-specific enolase (NSE) and S100B. Then, an in vitro blood-brain barrier (BBB) model, based on the human cerebral microvascular endothelial cell line (hCMEC/D3), was employed to assess the effect of plasma from the three study groups. Transendothelial electrical resistance (TEER) was used as an estimation of BBB integrity. NfL and tau are proteins expressed in axons, NSE in neurons and S100B in glial cells and are used as biomarkers for neurological injury in other diseases such as dementia, traumatic brain injury and hypoxic brain injury. Plasma concentrations of NfL, tau, NSE and S100B were all higher in women with preeclampsia compared with women with normal pregnancies (8.85 vs. 5.25 ng/L, p < 0.001; 2.90 vs. 2.40 ng/L, p < 0.05; 3.50 vs. 2.37 mu g/L, p < 0.001 and 0.08 vs. 0.05 mu g/L, p < 0.01, respectively). Plasma concentrations of NfL were also higher in women with preeclampsia compared with non-pregnant women (p < 0.001). Higher plasma concentrations of the cerebral biomarker NfL were associated with decreased TEER (p = 0.002) in an in vitro model of the BBB, a finding which indicates that NfL could be a promising biomarker for BBB alterations in preeclampsia., De två sista författarna delar sistaförfattarskapet.

Cerebral complications in preeclampsia contribute substantially to maternal mortality and morbidity. There is a lack of reliable and accessible predictors for preeclampsia-related cerebral complications. In this study, plasma from women with preeclampsia (n = 28), women with normal pregnancies (n = 28) and non-pregnant women (n = 16) was analyzed for concentrations of the cerebral biomarkers neurofilament light (NfL), tau, neuron-specific enolase (NSE) and S100B. Then, an in vitro blood-brain barrier (BBB) model, based on the human cerebral microvascular endothelial cell line (hCMEC/D3), was employed to assess the effect of plasma from the three study groups. Transendothelial electrical resistance (TEER) was used as an estimation of BBB integrity. NfL and tau are proteins expressed in axons, NSE in neurons and S100B in glial cells and are used as biomarkers for neurological injury in other diseases such as dementia, traumatic brain injury and hypoxic brain injury. Plasma concentrations of NfL, tau, NSE and S100B were all higher in women with preeclampsia compared with women with normal pregnancies (8.85 vs. 5.25 ng/L, p < 0.001; 2.90 vs. 2.40 ng/L, p < 0.05; 3.50 vs. 2.37 mu g/L, p < 0.001 and 0.08 vs. 0.05 mu g/L, p < 0.01, respectively). Plasma concentrations of NfL were also higher in women with preeclampsia compared with non-pregnant women (p < 0.001). Higher plasma concentrations of the cerebral biomarker NfL were associated with decreased TEER (p = 0.002) in an in vitro model of the BBB, a finding which indicates that NfL could be a promising biomarker for BBB alterations in preeclampsia., De två sista författarna delar sistaförfattarskapet.

Cerebral complications in preeclampsia contribute substantially to maternal mortality and morbidity. There is a lack of reliable and accessible predictors for preeclampsia-related cerebral complications. In this study, plasma from women with preeclampsia (n = 28), women with normal pregnancies (n = 28) and non-pregnant women (n = 16) was analyzed for concentrations of the cerebral biomarkers neurofilament light (NfL), tau, neuron-specific enolase (NSE) and S100B. Then, an in vitro blood-brain barrier (BBB) model, based on the human cerebral microvascular endothelial cell line (hCMEC/D3), was employed to assess the effect of plasma from the three study groups. Transendothelial electrical resistance (TEER) was used as an estimation of BBB integrity. NfL and tau are proteins expressed in axons, NSE in neurons and S100B in glial cells and are used as biomarkers for neurological injury in other diseases such as dementia, traumatic brain injury and hypoxic brain injury. Plasma concentrations of NfL, tau, NSE and S100B were all higher in women with preeclampsia compared with women with normal pregnancies (8.85 vs. 5.25 ng/L, p < 0.001; 2.90 vs. 2.40 ng/L, p < 0.05; 3.50 vs. 2.37 mu g/L, p < 0.001 and 0.08 vs. 0.05 mu g/L, p < 0.01, respectively). Plasma concentrations of NfL were also higher in women with preeclampsia compared with non-pregnant women (p < 0.001). Higher plasma concentrations of the cerebral biomarker NfL were associated with decreased TEER (p = 0.002) in an in vitro model of the BBB, a finding which indicates that NfL could be a promising biomarker for BBB alterations in preeclampsia., De två sista författarna delar sistaförfattarskapet.

Cerebral complications in preeclampsia contribute substantially to maternal mortality and morbidity. There is a lack of reliable and accessible predictors for preeclampsia-related cerebral complications. In this study, plasma from women with preeclampsia (n = 28), women with normal pregnancies (n = 28) and non-pregnant women (n = 16) was analyzed for concentrations of the cerebral biomarkers neurofilament light (NfL), tau, neuron-specific enolase (NSE) and S100B. Then, an in vitro blood-brain barrier (BBB) model, based on the human cerebral microvascular endothelial cell line (hCMEC/D3), was employed to assess the effect of plasma from the three study groups. Transendothelial electrical resistance (TEER) was used as an estimation of BBB integrity. NfL and tau are proteins expressed in axons, NSE in neurons and S100B in glial cells and are used as biomarkers for neurological injury in other diseases such as dementia, traumatic brain injury and hypoxic brain injury. Plasma concentrations of NfL, tau, NSE and S100B were all higher in women with preeclampsia compared with women with normal pregnancies (8.85 vs. 5.25 ng/L, p < 0.001; 2.90 vs. 2.40 ng/L, p < 0.05; 3.50 vs. 2.37 mu g/L, p < 0.001 and 0.08 vs. 0.05 mu g/L, p < 0.01, respectively). Plasma concentrations of NfL were also higher in women with preeclampsia compared with non-pregnant women (p < 0.001). Higher plasma concentrations of the cerebral biomarker NfL were associated with decreased TEER (p = 0.002) in an in vitro model of the BBB, a finding which indicates that NfL could be a promising biomarker for BBB alterations in preeclampsia., De två sista författarna delar sistaförfattarskapet.

Different strategies have been investigated for a more satisfactory treatment of advanced breast cancer, including the adjuvant use of omega-3 polyunsaturated fatty acids (PUFAs). These nutritional compounds have been shown to possess potent anti-inflammatory and antiangiogenic activities, the capacity to affect transduction pathways/receptors involved in cell growth and to reprogram tumor microenvironment. Omega-3 PUFA-containing nanoformulations designed for drug delivery in breast cancer were shown to potentiate the effects of enclosed drugs, enhance drug delivery to target sites, and minimize drug-induced side effects. We have critically analyzed here the results of the most recent studies investigating the effects of omega-3 PUFA-containing nanoformulations in breast cancer. The anti-neoplastic efficacy of omega-3 PUFAs has also been convincingly demonstrated by using preclinical in vivo models of ovarian cancer. The results obtained are critically analyzed here and seem to provide a sufficient rationale to move to still lacking interventional clinical trials, as well as to evaluate possible advantages of enclosing omega-3 PUFAs to drug-delivery nanosystems for ovarian cancer. Future perspectives in this area are also provided.

Cerebral complications in preeclampsia contribute substantially to maternal mortality and morbidity. There is a lack of reliable and accessible predictors for preeclampsia-related cerebral complications. In this study, plasma from women with preeclampsia (n = 28), women with normal pregnancies (n = 28) and non-pregnant women (n = 16) was analyzed for concentrations of the cerebral biomarkers neurofilament light (NfL), tau, neuron-specific enolase (NSE) and S100B. Then, an in vitro blood-brain barrier (BBB) model, based on the human cerebral microvascular endothelial cell line (hCMEC/D3), was employed to assess the effect of plasma from the three study groups. Transendothelial electrical resistance (TEER) was used as an estimation of BBB integrity. NfL and tau are proteins expressed in axons, NSE in neurons and S100B in glial cells and are used as biomarkers for neurological injury in other diseases such as dementia, traumatic brain injury and hypoxic brain injury. Plasma concentrations of NfL, tau, NSE and S100B were all higher in women with preeclampsia compared with women with normal pregnancies (8.85 vs. 5.25 ng/L, p < 0.001; 2.90 vs. 2.40 ng/L, p < 0.05; 3.50 vs. 2.37 mu g/L, p < 0.001 and 0.08 vs. 0.05 mu g/L, p < 0.01, respectively). Plasma concentrations of NfL were also higher in women with preeclampsia compared with non-pregnant women (p < 0.001). Higher plasma concentrations of the cerebral biomarker NfL were associated with decreased TEER (p = 0.002) in an in vitro model of the BBB, a finding which indicates that NfL could be a promising biomarker for BBB alterations in preeclampsia., De två sista författarna delar sistaförfattarskapet.

BACKGROUND: Cerebral complications in preeclampsia are leading causes of maternal mortality worldwide but the underlying pathophysiology is largely unknown and a challenge to study. Using an in vitro model of the human blood brain barrier (BBB), we explored the role of vascular endothelial growth factor receptor 2 (VEGFR2) in preeclampsia. METHODS: The human brain endothelial cell line (hCMEC/D3) cultured on Tranwells insert were exposed (12 h) to plasma from women with preeclampsia (n=28), normal pregnancy (n=28) and non-pregnant (n=16) controls. Transendothelial electrical resistance (TEER) and permeability to 70 kDa FITC-dextran were measured for assessment of BBB integrity. We explored possible underlying mechanisms, with focus on expression of tight junction proteins and phosphorylation of two tyrosine residues of VEGFR2, associated with vascular permeability and migration (pY951) and cell proliferation (pY1175). Plasma concentrations of soluble FMS like tyrosine kinase-1 (sFlt-1) and placental growth factor (PlGF) were measured in order to establish correlations with in vitro results. RESULTS: hCMEC/D3 exposed to plasma from women with preeclampsia exhibited reduced TEER and increased permeability to 70 kDa FITC-dextran. Further, these cells up-regulated the mRNA levels of VEGFR2, as well as pY951-VEGFR2; but reduced pY1175-VEGFR2 (p&0.05 in all cases). No difference in mRNA expression of tight junction protein was observed between gruops. There was no correlation between angiogenic biomarkers and BBB permeability. CONCLUSION: We present a promising in vitro model of the BBB in preeclampsia. Selective tyrosine phosphorylation of VEGFR2 may participate in the increased BBB permeability in preeclampsia irrespective of plasma concentrations of angiogenic biomarkers.

The β−-particle-emitting erbium-169 is a potential radionuclide toward therapy of metastasized cancer diseases. It can be produced in nuclear research reactors, irradiating isotopically-enriched 168Er2O3. This path, however, is not suitable for receptor-targeted radionuclide therapy, where high specific molar activities are required. In this study, an electromagnetic isotope separation technique was applied after neutron irradiation to boost the specific activity by separating 169Er from 168Er targets. The separation efficiency increased up to 0.5% using resonant laser ionization. A subsequent chemical purification process was developed as well as activity standardization of the radionuclidically pure 169Er. The quality of the 169Er product permitted radiolabeling and pre-clinical studies. A preliminary in vitro experiment was accomplished, using a 169Er-PSMA-617, to show the potential of 169Er to reduce tumor cell viability. © Copyright © 2021 Talip, Borgna, Müller, Ulrich, Duchemin, Ramos, Stora, Köster, Nedjadi, Gadelshin, Fedosseev, Juget, Bailat, Fankhauser, Wilkins, Lambert, Marsh, Fedorov, Chevallay, Fernier, Schibli and van der Meulen.

Aim. To evaluate in vitro the antibacterial effect of a paper made of silver phosphate cellulose fibers (SPCF) on Staphylococcus aureus, the most common diabetic foot ulceration (DFU) pathogen when compared with other common commercial products. Methods. The antibacterial activity of SPCF samples was evaluated through time with cell counting on agar plates. SPCF samples were then compared with commercial wound care products currently in use in DFU treatments (Silvercel™, Acticoat 7, and Aquacel Ag ExtraTM) through time on agar plates (growth inhibition zones). Results. After 6 hours, there was no viable bacterial cell detected on either plate (p < 0.05). There was a net growth inhibition zone for SPCF samples but no significant difference between the two silver concentrations. Compared with common commercial products, SPCF paper provides results equal to Acticoat 7 (p < 0.05) and superior to Aquacel AG ExtraTM and Silvercel™ at lower silver concentrations (p < 0.001). Conclusions. These results have shown the efficiency of SPCF paper to eliminate Staphylococcus aureus in these conditions. SPCF papers are effective when compared with other common commercial products and could have an industrial potential in wound care. Infected DFU could benefit from the antibacterial effectiveness of SPCF, but more relevant experimentations related to foot ulcers are needed.

Clinical guidance is often sought when prescribing drugs for patients with primary mitochondrial disease. Theoretical considerations concerning drug safety in patients with mitochondrial disease may lead to unnecessary withholding of a drug in a situation of clinical need. The aim of this study was to develop consensus on safe medication use in patients with a primary mitochondrial disease. A panel of 16 experts in mitochondrial medicine, pharmacology, and basic science from six different countries was established. A modified Delphi technique was used to allow the panellists to consider draft recommendations anonymously in two Delphi rounds with predetermined levels of agreement. This process was supported by a review of the available literature and a consensus conference that included the panellists and representatives of patient advocacy groups. A high level of consensus was reached regarding the safety of all 46 reviewed drugs, with the knowledge that the risk of adverse events is influenced both by individual patient risk factors and choice of drug or drug class. This paper details the consensus guidelines of an expert panel and provides an important update of previously established guidelines in safe medication use in patients with primary mitochondrial disease. Specific drugs, drug groups, and clinical or genetic conditions are described separately as they require special attention. It is important to emphasise that consensus-based information is useful to provide guidance, but that decisions related to drug prescribing should always be tailored to the specific needs and risks of each individual patient. We aim to present what is current knowledge and plan to update this regularly both to include new drugs and to review those currently included.

Clinical guidance is often sought when prescribing drugs for patients with primary mitochondrial disease. Theoretical considerations concerning drug safety in patients with mitochondrial disease may lead to unnecessary withholding of a drug in a situation of clinical need. The aim of this study was to develop consensus on safe medication use in patients with a primary mitochondrial disease. A panel of 16 experts in mitochondrial medicine, pharmacology, and basic science from six different countries was established. A modified Delphi technique was used to allow the panellists to consider draft recommendations anonymously in two Delphi rounds with predetermined levels of agreement. This process was supported by a review of the available literature and a consensus conference that included the panellists and representatives of patient advocacy groups. A high level of consensus was reached regarding the safety of all 46 reviewed drugs, with the knowledge that the risk of adverse events is influenced both by individual patient risk factors and choice of drug or drug class. This paper details the consensus guidelines of an expert panel and provides an important update of previously established guidelines in safe medication use in patients with primary mitochondrial disease. Specific drugs, drug groups, and clinical or genetic conditions are described separately as they require special attention. It is important to emphasise that consensus-based information is useful to provide guidance, but that decisions related to drug prescribing should always be tailored to the specific needs and risks of each individual patient. We aim to present what is current knowledge and plan to update this regularly both to include new drugs and to review those currently included.

Clinical guidance is often sought when prescribing drugs for patients with primary mitochondrial disease. Theoretical considerations concerning drug safety in patients with mitochondrial disease may lead to unnecessary withholding of a drug in a situation of clinical need. The aim of this study was to develop consensus on safe medication use in patients with a primary mitochondrial disease. A panel of 16 experts in mitochondrial medicine, pharmacology, and basic science from six different countries was established. A modified Delphi technique was used to allow the panellists to consider draft recommendations anonymously in two Delphi rounds with predetermined levels of agreement. This process was supported by a review of the available literature and a consensus conference that included the panellists and representatives of patient advocacy groups. A high level of consensus was reached regarding the safety of all 46 reviewed drugs, with the knowledge that the risk of adverse events is influenced both by individual patient risk factors and choice of drug or drug class. This paper details the consensus guidelines of an expert panel and provides an important update of previously established guidelines in safe medication use in patients with primary mitochondrial disease. Specific drugs, drug groups, and clinical or genetic conditions are described separately as they require special attention. It is important to emphasise that consensus-based information is useful to provide guidance, but that decisions related to drug prescribing should always be tailored to the specific needs and risks of each individual patient. We aim to present what is current knowledge and plan to update this regularly both to include new drugs and to review those currently included.

Aim. To evaluate in vitro the antibacterial effect of a paper made of silver phosphate cellulose fibers (SPCF) on Staphylococcus aureus, the most common diabetic foot ulceration (DFU) pathogen when compared with other common commercial products. Methods. The antibacterial activity of SPCF samples was evaluated through time with cell counting on agar plates. SPCF samples were then compared with commercial wound care products currently in use in DFU treatments (Silvercel™, Acticoat 7, and Aquacel Ag ExtraTM) through time on agar plates (growth inhibition zones). Results. After 6 hours, there was no viable bacterial cell detected on either plate (p < 0.05). There was a net growth inhibition zone for SPCF samples but no significant difference between the two silver concentrations. Compared with common commercial products, SPCF paper provides results equal to Acticoat 7 (p < 0.05) and superior to Aquacel AG ExtraTM and Silvercel™ at lower silver concentrations (p < 0.001). Conclusions. These results have shown the efficiency of SPCF paper to eliminate Staphylococcus aureus in these conditions. SPCF papers are effective when compared with other common commercial products and could have an industrial potential in wound care. Infected DFU could benefit from the antibacterial effectiveness of SPCF, but more relevant experimentations related to foot ulcers are needed.

N-(2-hydroxyphenyl)-1-[3-(2-oxo-2,3-dihydro-1H-benzimidazol -1-yl)propyl]piperidine-4-carboxamide (D2AAK4) is a multitarget ligand of aminergic G protein-coupled receptors (GPCRs) identified in structure-based virtual screening. Here we present detailed in vitro, in silico and in vivo investigations of this virtual hit. D2AAK4 has an atypical antipsychotic profile and low affinity to off-targets. It interacts with aminergic GPCRs, forming an electrostatic interaction between its protonatable nitrogen atom and the conserved Asp 3.32 of the receptors. At the dose of 100 mg/kg D2AAK4 decreases amphetamine-induced hyperactivity predictive of antipsychotic activity, improves memory consolidation in passive avoidance test and has anxiogenic properties in elevated plus maze test (EPM). Further optimization of the virtual hit D2AAK4 will be aimed to balance its multitarget profile and to obtain analogs with anxiolytic activity.

Clinical guidance is often sought when prescribing drugs for patients with primary mitochondrial disease. Theoretical considerations concerning drug safety in patients with mitochondrial disease may lead to unnecessary withholding of a drug in a situation of clinical need. The aim of this study was to develop consensus on safe medication use in patients with a primary mitochondrial disease. A panel of 16 experts in mitochondrial medicine, pharmacology, and basic science from six different countries was established. A modified Delphi technique was used to allow the panellists to consider draft recommendations anonymously in two Delphi rounds with predetermined levels of agreement. This process was supported by a review of the available literature and a consensus conference that included the panellists and representatives of patient advocacy groups. A high level of consensus was reached regarding the safety of all 46 reviewed drugs, with the knowledge that the risk of adverse events is influenced both by individual patient risk factors and choice of drug or drug class. This paper details the consensus guidelines of an expert panel and provides an important update of previously established guidelines in safe medication use in patients with primary mitochondrial disease. Specific drugs, drug groups, and clinical or genetic conditions are described separately as they require special attention. It is important to emphasise that consensus-based information is useful to provide guidance, but that decisions related to drug prescribing should always be tailored to the specific needs and risks of each individual patient. We aim to present what is current knowledge and plan to update this regularly both to include new drugs and to review those currently included.

Clinical guidance is often sought when prescribing drugs for patients with primary mitochondrial disease. Theoretical considerations concerning drug safety in patients with mitochondrial disease may lead to unnecessary withholding of a drug in a situation of clinical need. The aim of this study was to develop consensus on safe medication use in patients with a primary mitochondrial disease. A panel of 16 experts in mitochondrial medicine, pharmacology, and basic science from six different countries was established. A modified Delphi technique was used to allow the panellists to consider draft recommendations anonymously in two Delphi rounds with predetermined levels of agreement. This process was supported by a review of the available literature and a consensus conference that included the panellists and representatives of patient advocacy groups. A high level of consensus was reached regarding the safety of all 46 reviewed drugs, with the knowledge that the risk of adverse events is influenced both by individual patient risk factors and choice of drug or drug class. This paper details the consensus guidelines of an expert panel and provides an important update of previously established guidelines in safe medication use in patients with primary mitochondrial disease. Specific drugs, drug groups, and clinical or genetic conditions are described separately as they require special attention. It is important to emphasise that consensus-based information is useful to provide guidance, but that decisions related to drug prescribing should always be tailored to the specific needs and risks of each individual patient. We aim to present what is current knowledge and plan to update this regularly both to include new drugs and to review those currently included.

Aim. To evaluate in vitro the antibacterial effect of a paper made of silver phosphate cellulose fibers (SPCF) on Staphylococcus aureus, the most common diabetic foot ulceration (DFU) pathogen when compared with other common commercial products. Methods. The antibacterial activity of SPCF samples was evaluated through time with cell counting on agar plates. SPCF samples were then compared with commercial wound care products currently in use in DFU treatments (Silvercel™, Acticoat 7, and Aquacel Ag ExtraTM) through time on agar plates (growth inhibition zones). Results. After 6 hours, there was no viable bacterial cell detected on either plate (p < 0.05). There was a net growth inhibition zone for SPCF samples but no significant difference between the two silver concentrations. Compared with common commercial products, SPCF paper provides results equal to Acticoat 7 (p < 0.05) and superior to Aquacel AG ExtraTM and Silvercel™ at lower silver concentrations (p < 0.001). Conclusions. These results have shown the efficiency of SPCF paper to eliminate Staphylococcus aureus in these conditions. SPCF papers are effective when compared with other common commercial products and could have an industrial potential in wound care. Infected DFU could benefit from the antibacterial effectiveness of SPCF, but more relevant experimentations related to foot ulcers are needed.

Carbon monoxide has been demonstrated to exhibit several beneficial effects on biological targets (anti-inflammatory, anti-proliferative, anti-apoptotic effects, causes vasodilation, etc.).1 Consequently, the development of CO releasing molecules (CORMs), that allows for controlled release of CO under physiological conditions, has therefore become a major field of scientific and medical interest.2,3 Considerable research interest has been drawn on light-activated CORMs (photoCORMs) which only release CO upon radiation with a certain energy. However, despite a large number of photoCORMs reported, relatively little information is available on the precise mechanism of CO release from most photoCORMs and even less compounds have been tested as anti-cancer agents in cells so far. Herein, we report about the synthesis of ruthenium(II) dicarbonyl complexes functionalized with bidentate or tridentate (pyridyl, phenanthroline or diquinolyl) ligands and appending dyes for tuning the release properties as well as tracking the compound in cells. The mechanism of CO release in aqueous media (before and after light-activation) has been investigated. The photo-induced CO release kinetics of the Ru(II) photoCORMs, as well as in vitro studies in cancerous and healthy cell lines will be presented.4,5 References [1] R. Motterlini, L. E. Otterbein, Nat. Rev. Drug Discov. 9 (2010) 728-743. [2] U. Schatzschneider, Br. J. Pharmacol. 172 (2015) 1638-1650. [3] F. Zobi, Future Med. Chem. (2013) 5, 175-188. [4] M. Kubeil, R. R. Vernooij, C. Kubeil, B. R. Wood, B. Graham, H. Stephan, L. Spiccia, Inorg. Chem. 56 (2017) 5941-5952. [5] M. Kubeil, T. Joshi, B. R. Wood, H. Stephan, ChemistryOpen (2019) 8, 637-642. Acknowledgements MK was supported by a Marie Curie International Outgoing Fellowship from the European Union’s Seventh Framework Programme for research, technological development and demonstration under grant agreement no. 627113.

Carbon monoxide has been demonstrated to exhibit several beneficial effects on biological targets (anti-inflammatory, anti-proliferative, anti-apoptotic effects, causes vasodilation, etc.).1 Consequently, the development of CO releasing molecules (CORMs), that allows for controlled release of CO under physiological conditions, has therefore become a major field of scientific and medical interest.2,3 Considerable research interest has been drawn on light-activated CORMs (photoCORMs) which only release CO upon radiation with a certain energy. However, despite a large number of photoCORMs reported, relatively little information is available on the precise mechanism of CO release from most photoCORMs and even less compounds have been tested as anti-cancer agents in cells so far. Herein, we report about the synthesis of ruthenium(II) dicarbonyl complexes functionalized with bidentate or tridentate (pyridyl, phenanthroline or diquinolyl) ligands and appending dyes for tuning the release properties as well as tracking the compound in cells. The mechanism of CO release in aqueous media (before and after light-activation) has been investigated. The photo-induced CO release kinetics of the Ru(II) photoCORMs, as well as in vitro studies in cancerous and healthy cell lines will be presented.4,5 References [1] R. Motterlini, L. E. Otterbein, Nat. Rev. Drug Discov. 9 (2010) 728-743. [2] U. Schatzschneider, Br. J. Pharmacol. 172 (2015) 1638-1650. [3] F. Zobi, Future Med. Chem. (2013) 5, 175-188. [4] M. Kubeil, R. R. Vernooij, C. Kubeil, B. R. Wood, B. Graham, H. Stephan, L. Spiccia, Inorg. Chem. 56 (2017) 5941-5952. [5] M. Kubeil, T. Joshi, B. R. Wood, H. Stephan, ChemistryOpen (2019) 8, 637-642. Acknowledgements MK was supported by a Marie Curie International Outgoing Fellowship from the European Union’s Seventh Framework Programme for research, technological development and demonstration under grant agreement no. 627113.

The development of photo-activated carbon monoxide releasing molecules (photoCORMs) has received considerable attention as a new prodrug approach, since the CO is released only upon exposure to electromagnetic radiation [1]. Carbon monoxide itself has been demonstrated to exhibit several beneficial effects on biological targets such as anti-inflammation, anti-proliferation, anti-apoptosis, anti-oxidation and vasodilatory effects [2]. However, despite a large number of photoCORMs reported, relatively little information is available on the precise mechanism of CO release from most photoCORMs and even less compounds have been tested as anti-cancer agents in cells so far. Overall, a fundamental understanding of the mechanism of CO release from photoCORMs is essential for their exploitation as therapeutics. Herein, we report the synthesis of ruthenium(II) carbonyl complexes functionalized with bidentate pyridyl (1) and tridentate diquinolyl ligands (2) and investigate the mechanism of CO release in aqueous media (before and after light-activation). The photo-induced CO release kinetics of the Ru(II) photoCORMs, as well as the identity of the intermediates and photo-activated products, will be presented [3]. Moreover, the complexes have been tested in cancer cell lines showing a reduced viability after CO release. References [1] U. Schatzschneider, Br. J. Pharmacol. 2015, 172, 1638. [2] R. Motterlini, L. E. Otterbein, Nat. Rev. Drug Discov. 2010. 9, 728-743. [3] M. Kubeil, R. R. Vernooij, C. Kubeil, B. R. Wood, B. Graham, H. Stephan, L. Spiccia, Inorg. Chem. 2017, 56, 5941−5952.

The development of photo-activated carbon monoxide releasing molecules (photoCORMs) has received considerable attention as a new prodrug approach, since the CO is released only upon exposure to electromagnetic radiation [1]. Carbon monoxide itself has been demonstrated to exhibit several beneficial effects on biological targets such as anti-inflammation, anti-proliferation, anti-apoptosis, anti-oxidation and vasodilatory effects [2]. However, despite a large number of photoCORMs reported, relatively little information is available on the precise mechanism of CO release from most photoCORMs and even less compounds have been tested as anti-cancer agents in cells so far. Overall, a fundamental understanding of the mechanism of CO release from photoCORMs is essential for their exploitation as therapeutics. Herein, we report the synthesis of ruthenium(II) carbonyl complexes functionalized with bidentate pyridyl (1) and tridentate diquinolyl ligands (2) and investigate the mechanism of CO release in aqueous media (before and after light-activation). The photo-induced CO release kinetics of the Ru(II) photoCORMs, as well as the identity of the intermediates and photo-activated products, will be presented [3]. Moreover, the complexes have been tested in cancer cell lines showing a reduced viability after CO release. References [1] U. Schatzschneider, Br. J. Pharmacol. 2015, 172, 1638. [2] R. Motterlini, L. E. Otterbein, Nat. Rev. Drug Discov. 2010. 9, 728-743. [3] M. Kubeil, R. R. Vernooij, C. Kubeil, B. R. Wood, B. Graham, H. Stephan, L. Spiccia, Inorg. Chem. 2017, 56, 5941−5952.

Chitosan nanoparticles (CSNPs) ionically crosslinked with tripolyphosphate salts (TPP) were employedas nanocarriers in combined drug delivery and magnetic hyperthermia (MH) therapy. To that aim, threedifferent ferrofluid concentrations and a constant 5-fluorouracil (5-FU) concentration were efficientlyencapsulated to yield magnetic CSNPs with core-shell morphology. In vitro experiments using normalcells, fibroblasts (FHB) and cancer cells, human glioblastoma A-172, showed that CSNPs presented a dose-dependent cytotoxicity and that they were successfully uptaken into both cell lines. The application ofa MH treatment in A-172 cells resulted in a cell viability of 67–75% whereas no significant reduction ofcell viability was observed for FHB. However, the A-172 cells showed re-growth populations 4 h after theapplication of the MH treatment when CSNPs were loaded only with ferrofluid. Finally, a combined effectof MH and 5-FU release was observed with the application of a second MH treatment for CSNPs exhibitinga lower amount of released 5-FU. This result demonstrates the potential of CSNPs for the improvementof MH therapies

A potential complementary role of the dietary long-chain n-3 polyunsaturated fatty acids (LCn-3 PUFA) in combination with innovative mono-targeted therapies has recently been proposed. These compounds are thought to act pleiotropically to prevent the development and progression of a variety of cancers, including breast cancer. We hereinafter critically analyze the reports investigating the ability of LCn-3 PUFA to modulate the Ras/ERK and the phosphoinositide survival signaling pathways often aberrantly activated in breast cancer and representing the main targets of innovative therapies. The in vitro or in vivo animal and human interventional studies published up to January 2017 investigating the effects of LCn-3 PUFA on these pathways in normal and cancerous breast cells or tissues were identified through a systematic search of literature in the PubMed database. We found that, in most cases, both the in vitro and in vivo studies demonstrated the ability of LCn-3 PUFA to inhibit the activation of these pro-survival pathways. Altogether, the analyzed results strongly suggest a potential role of LCn-3 PUFA as complementary agents in combination with mono-targeted therapies. Moreover, the results indicate the need for further in vitro and human interventional studies designed to unequivocally prove the potential adjuvant role of these fatty acids.

During acute subarachnoid hemorrhage, blood is hemolyzed, which is followed by a significant cerebrovascular spasm resulting in a serious clinical condition. Interestingly, however, the direct vasomotor effect of perivascular hemolyzed blood (HB) has not yet been characterized, preventing the assessment of contribution of vasoconstrictor mechanisms deriving from brain tissue and/or blood and development of possible treatments. We hypothesized that perivascular HB reduces the diameter of the cerebral arteries (i.e., basilar artery [BA]; middle cerebral artery [MCA]) by elevating vascular tissue [Ca2+](i) level. Vasomotor responses were measured by videomicroscopy and intracellular Ca2+ by the Fura2-AM ratiometric method. Adding HB to the vessel chamber reduced the diameter significantly (BA: from 264 +/- 7 to 164 +/- 11 mu m; MCA: from 185 +/- 15 to 155 +/- 14 mu m), which was reversed to control level by wash-out of HB. Potassium chloride (KCl), HB, serum, hemolyzed red blood cell (RBC), plasma, and platelet suspension (PLTs) elicited significant constrictions of isolated basilar arteries. There was a significant increase in K+ concentration in hemolyzed HB (7.02 +/- 0.22 mmol/L) compared to Krebs' solution (6.20 +/- 0.01 mmol/L). Before HB, acetylcholine (ACh), sodium-nitroprussid (SNP), nifedipin, and CO2 elicited substantial dilations in cerebral arteries. In contrast, in the presence of HB dilations to ACh, SNP decreased, but not to nifedipine and CO2. After washout of HB, nitric oxide-mediated dilations remained significantly reduced compared to control. HB significantly increased the ratiometric Ca signal, which returned to control level after washout. In conclusion, perivascular hemolyzed blood elicits significant-nifedipine and high CO2 reversible-constrictions of isolated BAs and MCAs, primarily by increasing intracellular Ca2+, findings that can contribute to the refinement of local treatment of subarachnoid hemorrhage., Funding Agencies:American Heart AssociationOrszagos Tudomanyos Kutatasi AlapprogramokDeveloping Competitiveness of Universities in the South Transdanubian RegionHungarian Hypertension Society (MHT)

Through a process of translocation across biological barriers, nanoparticles can reach and deposit in secondary target organs where they may induce adverse biological reactions. Therefore, a correct assessment of nanoparticle-induced adverse effects should take into account the different aspects of toxicokinetics and tissues that may be targeted by nanoparticles. For this reason, a comprehensive evaluation of renal nanotoxicity is urgently needed as kidneys are particularly susceptible to xenobiotics and renal excretion is an expected and possible elimination route of nanoparticles in living organisms. On one hand, summarizing the findings of in vitro and in vivo studies that have investigated the adverse effects of nanoparticles on the kidney, this review intends to provide a thorough insight into the nephrotoxicity of these substances. The evaluation of the in vitro studies revealed that different types of nanoparticles (carbon, metal and/or silica nanoparticles) are able to exert significant cytotoxic effects (i.e., decreased cell viability, induction of oxidative stress, mitochondrial or cytoskeleton dysfunction and cell membrane and DNA damage). On the other hand, in vivo studies demonstrated that nanoparticles exhibited an important nephrotoxic potential both at tubular (i.e., degeneration of tubular epithelial cell, cellular fragments and proteinaceous liquid in tubule lumen, renal interstitial fibrosis) and glomerular level (i.e., swollen glomeruli, changes in Bowman's space and proliferation of mesangial cells). Although the data currently available indicate that nanoparticles may adversely impact the renal system, further studies are needed in order to clarify all the potential molecular mechanisms of nephrotoxicity induced by these xenobiotics, in particular at glomerular level.

The correct identification of immune-modulatory activity of polysaccharides is often hampered by immune-stimulatory contaminants, with pyrogens such as lipopolysaccharide (LPS) as a very potent example. In order to avoid false positive immuno-stimulatory properties to be attributed to polysaccharides, accurate quantification and inactivation of LPS in test samples is crucial. To quantify LPS in polysaccharide preparations of different origin and structure we used two different limulus amoebocyte lysate test kits in two different laboratories. We observed larger variation in detection of LPS contamination between kits than between labs. LPS quantification proved unreliable for some polysaccharide preparations as spike controls resulted in spike recoveries outside the acceptable range. We designed a cellular in vitro assay as alternative method to detect the presence of functional LPS. This HEK-Blue hTLR4 cell culture provides a reliable assay, when combined with a cell viability test, for determining functional LPS in polysaccharide preparations. Finally, to inactivate LPS in polysaccharide preparations, we setup an alkaline-ethanol-based treatment. With this assay we observed that our treatment (5 h incubation in 0.1 M NaOH) at 56 °C efficiently inactivated LPS in all polysaccharide preparations below immune-stimulatory levels. At this elevated temperature, however, we also observed minimal to severe degradation of polysaccharide preparations as determined with SEC-RI. Taken together, we describe methods and precautions to reliably detect and inactivate LPS in polysaccharide preparations to allow reliable in vitro investigations towards immune-modulatory potential of polysaccharide preparations.

The aqueous and hydro alcoholic extracts of Artemisia annua plants acclimatized and set in culture in Benin, were investigated for their in vitro and in vivo antimalarial value. These extracts were prepared by decoction and maceration using water and ethanol, and were tested for in vitro activity against the strains of Plasmodium falciparum. Those extracts were also investigated in vivo against Plasmodium berghei NK 173 infected mice by the 4-day suppressive test. The action of these extracts was similar in vitro to the action of pure artemisinin at the same dosage. But, the in vivo studies on mice show that we have the same efficacy with aqueous extract of A. annua (artemisinin content of 20 mg/kg) than with pure artemisinin at a dosage of 140 mg/kg. The hydro alcoholic extract of A. annua (artemisinin content of 20 mg/kg) was better than the both other conditions. The others components of the plant Artemisia annua potentiate artemisinin activity on Plasmodium. The aqueous and hydro alcoholic extracts of Artemisia annua L. are very effective on malaria and they can be used by populations to cure this disease, especially in endemic areas where modern medicines are expensive.

The study in this project was undertaken to document the effect of 24 hours exposure of cerium oxide (CeO2), cobalt oxide (Co3O4) and tungsten oxide (WO3) nanoparticles (NPs) on different cell lines. The study consisted of physicochemical characterization, cytotoxic, genotoxic and oxidative stress responses due to in vitro exposure to different doses of NPs and their microparticles (MPs). The physicochemical properties like size, size distribution, state of dispersion and zeta potential of these NPs are the important factors to study their toxicity. To investigate the potential role of oxidative stress as a mechanism of toxicity, reactive oxygen species (ROS), nitric oxide (NO), lactate dehydrogenase (LDH) level and reduction in the total glutathione (GSH) were estimated. The results showed that, there was significant induction of ROS, NO, LDH and GSH level observed in IMR32 cells treated with CeO2 NPs from 100 g/ml onwards, whereas in their bulk no significant changes were observed. Similarly, HEK293 cells treated with Co3O4 NPs showed significant increase in ROS, membrane leakage of LDH and NO and reduction in the total GSH at the dose level of 50 g/ml onwards however, bulk were not found to induce any significant changes. Comet assay of the NMs and bulk treated cells showed a concentration dependent increase in the percentage of tail DNA, however only the NMs showed a significant increase.

A study on the physicochemical properties and MTT cytotoxicity assay of cerium oxide (CeO2), cobalt oxide (Co3O4) and tungsten oxide (WO3) nanomaterials (NMs) on different cell lines was carried out during the last year. The physicochemical properties like size, size distribution, state of dispersion and zeta potential of these NMs are the important factors to study their toxicity.

The derivatives of cinnamic acid and N-arylpiperazine show antibacterial activity. In this work the potential synergistic effect of cinnamyl derivatives of arylpiperazine in selected bacteria was investigated. The antibacterial activities of the derivatives were evaluated against Gram-positive bacteria: Staphylococcus aureus, Streptosporangium longisporum, Sarcina lutea, Micrococcus flavus, Clostridium sporogenes and Bacillus subtilis and Gram-negative bacteria: Escherichia coli, Pseudomonas aeruginosa, Salmonella enteritidis and Proteus vulgaris by the disc diffusion method. The minimum inhibitory concentration (MIC) against the selected bacteria was determined for all compounds that showed activity in the disc diffusion method. The majority of the investigated compounds displayed in vitro antibacterial activity. The effect of the type and structure of the substituent on the aromatic ring on the antibacterial activity is discussed. It was found that two derivatives expressed activity toward S. longisporum and P. aeruginosa that was almost as strong as that of amikacin.

The derivatives of cinnamic acid and N-arylpiperazine show antibacterial activity. In this work the potential synergistic effect of cinnamyl derivatives of arylpiperazine in selected bacteria was investigated. The antibacterial activities of the derivatives were evaluated against Gram-positive bacteria: Staphylococcus aureus, Streptosporangium longisporum, Sarcina lutea, Micrococcus flavus, Clostridium sporogenes and Bacillus subtilis and Gram-negative bacteria: Escherichia coli, Pseudomonas aeruginosa, Salmonella enteritidis and Proteus vulgaris by the disc diffusion method. The minimum inhibitory concentration (MIC) against the selected bacteria was determined for all compounds that showed activity in the disc diffusion method. The majority of the investigated compounds displayed in vitro antibacterial activity. The effect of the type and structure of the substituent on the aromatic ring on the antibacterial activity is discussed. It was found that two derivatives expressed activity toward S. longisporum and P. aeruginosa that was almost as strong as that of amikacin.

The early identification of the insurgence of a malignant cancer and the selective targeting of the tumor with specific drugs are still an open frontier for cancer diagnosis and treatment. The ultimate goal is to improve the therapy efficiency and to reduce the side effects usually encountered with conventional chemotherapy. Worldwide, mammary carcinoma represents the second most recurrent type of malignant tumor in adult women and the fifth cause of death among cancer types. In the context of this thesis, I have designed and developed multifunctional hybrid nanoparticles consisting of an inorganic iron oxide core, useful as source of signal for magnetic resonance imaging (MRI), and an organic shell, including bioactive ligands for the pharmacological effect combined with specific cell targeting, and a molecular dye as fluorescence signal emitter. The nanoparticle characteristics were optimized in terms of size, morphology, surface charge, stability, fluorescence emission and capability to enhance the MRI contrast. In addition, specific biomolecular ligands based on anti-HER-2 monoclonal antibody have been developed and novel strategies for their conjugation to nanoparticles were explored. The resulting hybrid nanocomplexes were tested both in vitro and in vivo to evaluate their toxicity, endocytosis, degradation pathways, and the efficient recognition of cell-surface biomarkers. Next, these nanoparticles proved to be highly effective in selectively targeting breast cancer cells in transplanted mice bearing HER-2-positive tumors. A multifaceted bioanalytical approach, combining fluorescence, magnetic relaxivity, transmission electron microscopy, and histological experiments in vivo and ex vivo, has demonstrated that these nanoprobes prevalently accumulated at the tumor by an active targeting route. The nanoparticles were endocytosed by the tumor cells following a lysosomal pathway of degradation, while did not result in permanent damage of healthy tissues. The princ, CORSI, FABIO, 1692, open, open, Colombo, Colombo, M

The derivatives of cinnamic acid and N-arylpiperazine show antibacterial activity. In this work the potential synergistic effect of cinnamyl derivatives of arylpiperazine in selected bacteria was investigated. The antibacterial activities of the derivatives were evaluated against Gram-positive bacteria: Staphylococcus aureus, Streptosporangium longisporum, Sarcina lutea, Micrococcus flavus, Clostridium sporogenes and Bacillus subtilis and Gram-negative bacteria: Escherichia coli, Pseudomonas aeruginosa, Salmonella enteritidis and Proteus vulgaris by the disc diffusion method. The minimum inhibitory concentration (MIC) against the selected bacteria was determined for all compounds that showed activity in the disc diffusion method. The majority of the investigated compounds displayed in vitro antibacterial activity. The effect of the type and structure of the substituent on the aromatic ring on the antibacterial activity is discussed. It was found that two derivatives expressed activity toward S. longisporum and P. aeruginosa that was almost as strong as that of amikacin.

The derivatives of cinnamic acid and N-arylpiperazine show antibacterial activity. In this work the potential synergistic effect of cinnamyl derivatives of arylpiperazine in selected bacteria was investigated. The antibacterial activities of the derivatives were evaluated against Gram-positive bacteria: Staphylococcus aureus, Streptosporangium longisporum, Sarcina lutea, Micrococcus flavus, Clostridium sporogenes and Bacillus subtilis and Gram-negative bacteria: Escherichia coli, Pseudomonas aeruginosa, Salmonella enteritidis and Proteus vulgaris by the disc diffusion method. The minimum inhibitory concentration (MIC) against the selected bacteria was determined for all compounds that showed activity in the disc diffusion method. The majority of the investigated compounds displayed in vitro antibacterial activity. The effect of the type and structure of the substituent on the aromatic ring on the antibacterial activity is discussed. It was found that two derivatives expressed activity toward S. longisporum and P. aeruginosa that was almost as strong as that of amikacin.

The derivatives of cinnamic acid and N-arylpiperazine show antibacterial activity. In this work the potential synergistic effect of cinnamyl derivatives of arylpiperazine in selected bacteria was investigated. The antibacterial activities of the derivatives were evaluated against Gram-positive bacteria: Staphylococcus aureus, Streptosporangium longisporum, Sarcina lutea, Micrococcus flavus, Clostridium sporogenes and Bacillus subtilis and Gram-negative bacteria: Escherichia coli, Pseudomonas aeruginosa, Salmonella enteritidis and Proteus vulgaris by the disc diffusion method. The minimum inhibitory concentration (MIC) against the selected bacteria was determined for all compounds that showed activity in the disc diffusion method. The majority of the investigated compounds displayed in vitro antibacterial activity. The effect of the type and structure of the substituent on the aromatic ring on the antibacterial activity is discussed. It was found that two derivatives expressed activity toward S. longisporum and P. aeruginosa that was almost as strong as that of amikacin.

Increasing evidence indicates that tomato lycopene may be an ideal candidate in protecting from cancer risk related to smoking exposure. The carotenoid shows potent redox-properties by which it decreases the reactive radical species (ROS) generated by smoke and modulates redox-sensitive cell targets, including protein tyrosine phosphatases, protein kinases, MAPKs and transcription factors. Moreover, it counteracts the effects of smoke on carcinogen-bioactivating enzymes and on molecular pathways involved in cell proliferation, apoptosis and inflammation. Lycopene also inhibits smoke-stimulated IGF-signalling and smoke-induced DNA adducts. Some of these actions may be mediated by its oxidative metabolites and may be synergistically enhanced by the presence of other antioxidant nutrients. This review summarizes the background information about the interactions of lycopene with smoke in experimental models and presents the most current knowledge with respect to lycopene role in smoke-related diseases.

A huge amount of different nanomaterials is nowadays on the market used for various specific applications. Some nanomaterials such as TiO2, ZnO as well as several other semiconductors exhibit photocatalytic activity. Hence these materials are used for many applications, e.g., for self-cleaning and antibacterial coatings on different surfaces and for the purification of wastewater where the cleaning can be induced by simple exposure to sunlight. Because of the frequent use of these nanoparticles it is important to investigate the life cycles of these nanostructured materials as well as their environmental impact and their toxicity to animals and humans. This review first gives a short overview about nanotechnology and nanotechnological products as well as about photocatalysis and semiconductors used in this field. We then discuss the need for a new technology named nanotoxicology and the problems occurring when investigating the toxic potential of nanomaterials as well as the life cycle of nanomaterials. Furthermore, we focus on the environmental impact of TiO2 and ZnO nanoparticles including toxic effects to bacteria, water organisms and plants as well as their toxic effects to humans including in vitro and in vivo studies.

Lycopene, a natural carotenoid found in tomato, has been reported to possess various health benefits, such as cardiovascular and cancer preventive properties. However, the experimental basis for such health benefits is not fully understood. One of the possible mechanisms for its protective activities is by down-regulation of the inflammatory response. That includes the inhibition of pivotal pro-inflammatory mediators, such as the reduction of reactive oxygen species, the inhibition of synthesis and release of pro-inflammatory cytokines, changes in the expression of cyclooxygenase and lipoxygenase, modifications of eicosanoid synthesis, and modulation of signal transduction pathways, including that of the inducible nitric oxide synthase via its inhibitory effects on Nuclear Factor-kB (NF-kB), Activated protein-1 (AP-1) and mitogen-activated protein kinase (MAPK) signaling. Recent data suggest that lycopene also exhibits anti-inflammatory activity through induction of programmed cell death in activated immune cells. This review will discuss recent data on the control of inflammatory signaling exerted by tomato lycopene in isolated cells, in animal models and in clinical trials, focusing on the dose of the carotenoid and the biological environment in which it acts. A clear understanding of the molecular mechanisms of action of lycopene is crucial in the valuation of this molecule as a potential preventive and therapeutic agent.

In order to minimize the risk of contamination associated with tissue transplantation, tissue banks commonly chemically treat the tissues whenever possible. As viral inactivation uses agents lethal to microorganisms, it is imperative to assure that chemically inactivated tissue remains biocompatible. In vitro assays can be an effective means to assess the acute cytotoxicity of chemically treated human allografts. We have used different types of cells cultured in the presence of treated tissue extract. A standard cell line, a human fibroblast (WI38), which was the same for all the samples, was chosen. In addition, as the banked tissues (bone and fascia lata) were prepared to be used in bone or as a dura mater substitute, two other cell types were also used: an osteoblastic cell line (SaOS-2) and a neuronal cell line (Neuro 2A). Cytotoxic assessment was performed by qualitative evaluation of cell morphology based on confluence, granulation, vacuolization and swelling analysis. In addition, quantitative methods based on the use of neutral red (NR) and 3- (4,5-dimethylthiazol-2-yl)-2,5 diphenyl tetrazolium bromide (MTT) were assayed. Qualitative and quantitative evaluation of fascia lata and bone extracts did not show deleterious effects on cell cultures. These results show that in vitro methods can be appropriate to select a non-toxic procedure before it is used in the human body and that several strong chemical treatments can result in a tissue suitable for human.

Imatinib mesylate has antitumor activity in vitro and in vivo against neuroblastoma cell lines and xenografts characterized by a different expression of receptor tyrosine kinases. In this article, we report that imatinib tumor concentration can be independent of the administered dose and does not correlate with the antitumor effect. In xenografts, high-dose administration does not improve imatinib efficacy. In conclusion, there is no clear-cut correlation between the levels of expression for imatinib-responsive targets and the in vitro and in vivo sensitivity. This further suggests that in neuroblastoma the antitumor activity of imatinib may involve the inhibition of other tyrosine kinases and/or pathways.

We studied the androgen receptor gene in a large kindred with complete androgen insensitivity syndrome and negative receptor-binding activity, single-strand conformation polymorphism (SSCP) analysis and sequencing identified a 13 base pair deletion within exon 4. This was responsible for a predictive frameshift in the open reading frame and introduction of a premature stop codon at position 783 instead of 919. The deletion was reproduced in androgen receptor wildtype cDNA and transfected into mammalian cells. Western blot showed a smaller androgen receptor of 94 kDa for the transfected mutated cDNA instead of 110 kDa. Androgen-binding assay of the mutated transfected cells assessed the lack of androgen-binding. Gel retardation assay demonstrated the ability of the mutant to bind target DNA; however, the mutant was unable to transactivate a reporter gene. Although the role of the partial deletion in the lack of androgen action was expected, in vitro analyses highlight the role of the abnormal C-terminal portion in the inhibition of the receptor transregulatory activity of the protein causing androgen resistance in this family.