Novel D-modified heterocyclic androstane derivatives as potential anticancer agents: Synthesis, characterization, in vitro and in silico studies.

Cancer remains a major health concern worldwide. The most frequently diagnosed types of cancer are caused by abnormal production or action of steroid hormones. In the present study, the synthesis and structural characterization of new heterocyclic androstane derivatives with D-homo lactone, 17α-(pyridine-2''-ylmethyl) or 17(E)-(pyridine-2''-ylmethylidene) moiety are presented. All compounds were evaluated for their anti-proliferative activity against HeLa cervical cancer cell line and non-cancerous kidney MDCK cells, where A-homo lactam compound 9A showed the greatest selectivity. Based on in vitro binding assays, N-formyl lactam compound 18 appeared to be the strong and isoform-selective ligand for ERα, while compound 9A displayed binding affinity for the GR-LBD, but also inhibited aldo-keto reductase 1C4 enzyme. Out of four selected compounds, methylpyrazolo derivative 13 showed potential for aromatase binding, while in silico studies provided insight into experimentally confirmed protein-ligand interactions.

Lyophilized Human Bone Allograft as an Antibiotic Carrier: An In Vitro and In Vivo Study.

Background: Antibiotics delivered from implanted bone substitute materials (BSM) can potentially be used to prevent acute infections and biofilm formation, providing high concentrations of antibiotics at the surgical site without systemic toxicity. In addition, BSM should allow osteoconductivity supporting bone healing without further surgery. Promising results have been achieved using lyophilized bone allografts mixed with antibiotics. Methods: In this study specially prepared human bone allografts were evaluated as an antibiotic carrier in vitro and in vivo. The efficacy of different antibiotic-impregnated bone allografts was measured by drug release tests in vitro and in vivo and bacterial susceptibility tests using four bacterial species usually responsible for implant-associated infections. Results: The loading procedures of allograft bone substitutes with antibiotics were successful. Some of the antibiotic concentrations exceeded the MIC90 for up to 7 days in vitro and for up to 72 h in vivo. The susceptibility tests showed that S. epidermidis ATCC 12228 was the most susceptible bacterial species in comparison to the other strains tested for all antibiotic substances. Vancomycin and rifampicin showed the best results against standard and patient-isolated strains in vitro. In vivo, new bone formation was comparable in all study groups including the control group without antibiotic loading. Conclusions: Human bone allografts showed the capacity to act as customized loaded antibiotic carriers to prevent acute infections and should be considered in the management of bone infections in combination with systemic antimicrobial therapy.

Mucinous cystadenoma of the liver with pathological-radiological correlation.

The paper presents a pathological-radiological correlation of the manifestation of mucosal cystadenoma with ovarian stroma of the liver with examination and correlation with the new stroma nomenclature and differential diagnostic dilemmas of radiologists and pathologists.

How to improve O-RADS MRI score for rating adnexal masses with cystic component?

OBJECTIVES: To test the performance of the Ovarian-Adnexal Reporting Data System (O-RADS) MRI in characterizing adnexal masses with cystic components and to test new specific MRI features related to cystic components to improve the ability of the O-RADS MRI score to stratify lesions according to their risk of malignancy. METHODS: The EURopean ADnexal study (EURAD) database was retrospectively queried to identify adnexal masses with a cystic component. One junior and 13 radiologists independently reviewed cases blinded to the pathological diagnosis. For each lesion, the size of the whole lesion, morphological appearance, number of loculi, presence of a thickened wall, thickened septae, signal intensity of the cystic components on T1-weighted/T2-weighted/diffusion weighted, mean value of the apparent diffusion coefficient, and O-RADS MRI score were reported. Univariate and multivariate logistic regression analysis was performed to determine significant features to predict malignancy. RESULTS: The final cohort consisted of 585 patients with 779 pelvic masses who underwent pelvic MRI to characterize an adnexal mass(es). Histology served as the standard of reference. The diagnostic performance of the O-RADS MRI score was 0.944, 95%CI [0.922-0.961]. Significant criteria associated with malignancy included an O-RADS MRI score ≥ 4, ADCmean of cystic component > 1.69, number of loculi > 3, lesion size > 75 mm, the presence of a thick wall, and a low T1-weighted, a high T2-weighted, and a low diffusion-weighted signal intensity of the cystic component. Multivariate analysis demonstrated that an O-RADS MRI score ≥ combined with an ADC mean of the cystic component > 1.69, size > 75 mm, and low diffusion-weighted signal of the cystic component significantly improved the diagnostic performance up to 0.958, 95%CI [0.938-0.973]. CONCLUSION: Cystic component analysis may improve the diagnosis performance of the O-RADS MRI score in adnexal cystic masses. KEY POINTS: • O-RADS MRI score combined with specific cystic features (area under the receiving operating curve, AUROC = 0.958) improves the diagnostic performance of the O-RADS MRI score (AUROC = 0.944) for predicting malignancy in this cohort. • Cystic features that improve the prediction of malignancy are ADC mean > 1.69 (OR = 7); number of loculi ≥ 3 (OR = 5.16); lesion size > 75 mm (OR = 4.40); the presence of a thick wall (OR = 3.59); a high T2-weighted signal intensity score 4 or 5 (OR = 3.30); a low T1-weighted signal intensity score 1, 2, or 3 (OR = 3.45); and a low diffusion-weighted signal intensity (OR = 2.12). • An adnexal lesion with a cystic component rated O-RADS MRI score 4 and an ADC value of the cystic component < 1.69 associated with a low diffusion-weighted signal, has virtually a 0% risk of malignancy.

In Vivo Biocompatibility Investigation of an Injectable Calcium Carbonate (Vaterite) as a Bone Substitute including Compositional Analysis via SEM-EDX Technology.

Injectable bone substitutes (IBS) are increasingly being used in the fields of orthopedics and maxillofacial/oral surgery. The rheological properties of IBS allow for proper and less invasive filling of bony defects. Vaterite is the most unstable crystalline polymorph of calcium carbonate and is known to be able to transform into hydroxyapatite upon contact with an organic fluid (e.g., interstitial body fluid). Two different concentrations of hydrogels based on poly(ethylene glycol)-acetal-dimethacrylat (PEG-a-DMA), i.e., 8% (w/v) (VH-A) or 10% (w/v) (VH-B), were combined with vaterite nanoparticles and implanted in subcutaneous pockets of BALB/c mice for 15 and 30 days. Explants were prepared for histochemical staining and immunohistochemical detection methods to determine macrophage polarization, and energy-dispersive X-ray analysis (EDX) to analyze elemental composition was used for the analysis. The histopathological analysis revealed a comparable moderate tissue reaction to the hydrogels mainly involving macrophages. Moreover, the hydrogels underwent a slow cellular infiltration, revealing a different degradation behavior compared to other IBS. The immunohistochemical detection showed that M1 macrophages were mainly found at the material surfaces being involved in the cell-mediated degradation and tissue integration, while M2 macrophages were predominantly found within the reactive connective tissue. Furthermore, the histomorphometrical analysis revealed balanced numbers of pro- and anti-inflammatory macrophages, demonstrating that both hydrogels are favorable materials for bone tissue regeneration. Finally, the EDX analysis showed a stepwise transformation of the vaterite particle into hydroxyapatite. Overall, the results of the present study demonstrate that hydrogels including nano-vaterite particles are biocompatible and suitable for bone tissue regeneration applications.

Standardized wild apple fruit extract as a bioactive agent in dermocosmetic products for efficacy skin hydration-In vitro and in vivo evaluation.

BACKGROUND: Wild apple fruit is rich source of bioactive substances, which can be potentially used for a production of dermocosmetic products. AIM: The aim of study was in vitro and in vivo investigation of standardized wild apple fruit extracts (obtained by purified water and maceration-EM, percolation-EP, Soxhlet extraction-ES and ultrasonic extraction-EU) and dermocosmetic creams of o/w type containing 6%, 12%, and 15% of extract EU, as extract with the best content of polyphenolic compounds (CE6, CE12, and CE15, respectively), in order to determine the optimal concentration of the extract, which manifests the best effect on the biophysical characteristics of the skin after application. METHODS: Content of polyphenolic substances-PP was investigated using HPLC analysis, while in vitro biological activity was examined on L929 fibroblasts using MTT viability test. In vivo efficacy included screening and comparison of skin hydration potential, transepidermal water loss-TEWL, and skin pH after 28 days of creams application on healthy volunteers' skin. RESULTS: Content of identified PP was the highest in the extract EU (2001.57 ± 0.87 mg PP/100 g EU) and satisfactory in cream CE6 (53.83 ± 0.23 mg PP/100 g CE6). Procyanidin B2 was the most common PP. Biological activity analysis showed that extract EU could be considered as non-cytotoxic for fibroblasts in examined concentrations. In vivo investigation revealed an increase of skin hydration (after 28 days ΔEC was 21.19 ± 7.59, 26.71 ± 7.94, and 29.60 ± 10.95 for CE6, CE12, and CE15, respectively), with unchanged TEWL and skin pH values. CONCLUSION: Obtained results indicate that wild apple fruit extract represents a valuable source of bioactive substances, with good hydration effects of creams on human skin (effect of creams increased proportionally to the increase of concentration of incorporated extract), so it might be considered as a great potential for application in dermocosmetic industry.

Biocompatibility Analyses of HF-Passivated Magnesium Screws for Guided Bone Regeneration (GBR).

Background: Magnesium (Mg) is one of the most promising materials for human use in surgery due to material characteristics such as its elastic modulus as well as its resorbable and regenerative properties. In this study, HF-coated and uncoated novel bioresorbable magnesium fixation screws for maxillofacial and dental surgical applications were investigated in vitro and in vivo to evaluate the biocompatibility of the HF coating. Methods: Mg alloy screws that had either undergone a surface treatment with hydrofluoric-acid (HF) or left untreated were investigated. In vitro investigation included XTT, BrdU and LDH in accordance with the DIN ISO 10993-5/-12. In vivo, the screws were implanted into the tibia of rabbits. After 3 and 6 weeks, degradation, local tissue reactions and bony integration were analyzed histopathologically and histomorphometrically. Additionally, SEM/EDX analysis and synchrotron phase-contrast microtomography (µCT) measurements were conducted. The in vitro analyses revealed that the Mg screws are cytocompatible, with improved results when the surface had been passivated with HF. In vivo, the HF-treated Mg screws implanted showed a reduction in gas formation, slower biodegradation and a better bony integration in comparison to the untreated Mg screws. Histopathologically, the HF-passivated screws induced a layer of macrophages as part of its biodegradation process, whereas the untreated screws caused a slight fibrous tissue reaction. SEM/EDX analysis showed that both screws formed a similar layer of calcium phosphates on their surfaces and were surrounded by bone. Furthermore, the µCT revealed the presence of a metallic core of the screws, a faster absorbing corrosion front and a slow absorbing region of corroded magnesium. Conclusions: Overall, the HF-passivated Mg fixation screws showed significantly better biocompatibility in vitro and in vivo compared to the untreated screws.

Macrophages' contribution to ectopic osteogenesis in combination with blood clot and bone substitute: possibility for application in bone regeneration strategies.

PURPOSE: Given the great potential of macrophages in the processes of tissue repair and regeneration, the aim of our study was to examine the contribution that macrophages will have in osteogenic process when combined and implanted with blood clot (BC) and mineral bone substitute (MBS) in mice subcutaneous implantation model. METHODS: Three types of implants were constructed and implanted subcutaneously into BALB/c mice: (1) RMBM implants (made of resident tissue macrophages, BC and MBS), (2) BM implants (made of BC and MBS), and (3) M implants (made of MBS only) where the last two served as control implants. One, two, four and eight weeks after implantation implants were explanted, and histochemical, immunohistochemical, and histomorphometric analyses were performed. RESULTS: Increased vascularization, particularly pronounced two and four weeks after implantation and pronounced tissue infiltration in eight week term in RMBM implants compared with both other types, likewise the presence of osteoblast-like cells, osteoid-like structures, and more prominent osteopontin and osteocalcin immunoexpression in RMBM implants indicated more pronounced osteogenic process within them. CONCLUSION: Our results suggest that macrophages deserve to be considered as a cell component when constructing implants in bone regenerative medicine strategies to improve bone fracture healing process.

In Vitro and In Vivo Biocompatibility Analysis of a New Transparent Collagen-based Wound Membrane for Tissue Regeneration in Different Clinical Indications.

BACKGROUND/AIM: For the treatment of different tissue defects such as jawbone defects, open wound defect, chronic ulcers, dura mater defects and corneal defects, different biomaterials are available. The use of collagen-based materials for these applications has been significantly increased over the past decades due to its excellent biocompatibility and degradability. However, no transparent collagen-based biomaterial is available until now. Thus, a newly developed transparent collagen membrane (TCM) based on natural derived porcine pericardium, which offers numerous application possibilities, was developed. The present study aimed to analyze the in vitro and in vivo biocompatibility using established methods. MATERIALS AND METHODS: The new TCM membrane and a commercially available collagen membrane (CM, Jason membrane, botiss biomaterials GmbH, Zossen, Germany) were tested for its in vitro cytocompatibility. Furthermore, the in vivo biocompatibility was analyzed using sham operations as control group. In vitro, cytocompatibility was tested in accordance with EN ISO 10993-5/-12 regulations and Live-Dead-stainings. In vivo, a subcutaneous implantation model in BALB/c mice was used and explants were prepared for analyses by established histological, immunohistochemical and histomorphometrical methods. RESULTS: In vitro, both membranes showed promising cytocompatibility with a slightly better direct cell response in the Live-Dead staining assay for the TCM. In vivo, TCM induced a comparable inflammatory immune response after 10 and 30 days with comparable numbers of M1- and M2-macrophages as also found in the control group without biomaterial insertion. CONCLUSION: The newly transparent collagen membrane is fully biocompatible and is supporting safe clinical application in tissue repair and surgery.

The Condensation of Collagen Leads to an Extended Standing Time and a Decreased Pro-inflammatory Tissue Response to a Newly Developed Pericardium-based Barrier Membrane for Guided Bone Regeneration.

BACKGROUND/AIM: A new manufacturing process has been established for the condensation of collagen derived from porcine pericardium to develop a new dental barrier membrane (CPM) that can provide a long barrier functionality. A native collagen membrane (PM) was used as control. MATERIALS AND METHODS: Established in vitro procedures using L929 and MC3T3 cells were used for cytocompatibility analyses. For the in vivo study, subcutaneous implantation of both membrane types in 40 BALB/c mice and established histological, immuno histochemical and histomorphometrical methods were conducted. RESULTS: Both the in vitro and in vivo results revealed that the CPM has a biocompatibility profile comparable to that of the control membrane. The new CPM induced a tissue reaction including more M2-macrophages. CONCLUSION: The CPM is fully biocompatible and seems to support the early healing process. Moreover, the new biomaterial seems to prevent cell ingrowth for a longer period of time, making it ideally suited for GBR procedures.

Studies on the Mechanisms of Anti-Inflammatory Activity of Heparin- and Hyaluronan-Containing Multilayer Coatings-Targeting NF-κB Signalling Pathway.

The use of implants can be hampered by chronic inflammatory reactions, which may result in failure of the implanted device. To prevent such an outcome, the present study examines the anti-inflammatory properties of surface coatings made of either hyaluronic acid (HA) or heparin (Hep) in combination with chitosan (Chi) prepared as multilayers through the layer-by-layer (LbL) technique. The properties of glycosaminoglycan (GAG)-modified surfaces were characterized in terms of surface topography, thickness and wettability. Results showed a higher thickness and hydrophilicity after multilayer formation compared to poly (ethylene imine) control samples. Moreover, multilayers containing either HA or Hep dampened the inflammatory response visible by reduced adhesion, formation of multinucleated giant cells (MNGCs) and IL-1ß release, which was studied using THP-1 derived macrophages. Furthermore, investigations regarding the mechanism of anti-inflammatory activity of GAG were focused on nuclear transcription factor-кB (NF-κB)-related signal transduction. Immunofluorescence staining of the p65 subunit of NF-κB and immunoblotting were performed that showed a significant decrease in NF-κB level in macrophages on GAG-based multilayers. Additionally, the association of FITC-labelled GAG was evaluated by confocal laser scanning microscopy and flow cytometry showing that macrophages were able to associate with and take up HA and Hep. Overall, the Hep-based multilayers demonstrated the most suppressive effect making this system most promising to control macrophage activation after implantation of medical devices. The results provide an insight on the anti-inflammatory effects of GAG not only based on their physicochemical properties, but also related to their mechanism of action toward NF-κB signal transduction.

Ovarian-Adnexal Reporting Data System Magnetic Resonance Imaging (O-RADS MRI) Score for Risk Stratification of Sonographically Indeterminate Adnexal Masses.

Importance: Approximately one-quarter of adnexal masses detected at ultrasonography are indeterminate for benignity or malignancy, posing a substantial clinical dilemma. Objective: To validate the accuracy of a 5-point Ovarian-Adnexal Reporting Data System Magnetic Resonance Imaging (O-RADS MRI) score for risk stratification of adnexal masses. Design, Setting, and Participants: This multicenter cohort study was conducted between March 1, 2013, and March 31, 2016. Among patients undergoing expectant management, 2-year follow-up data were completed by March 31, 2018. A routine pelvic MRI was performed among consecutive patients referred to characterize a sonographically indeterminate adnexal mass according to routine diagnostic practice at 15 referral centers. The MRI score was prospectively applied by 2 onsite readers and by 1 reader masked to clinical and ultrasonographic data. Data analysis was conducted between April and November 2018. Main Outcomes and Measures: The primary end point was the joint analysis of true-negative and false-negative rates according to the MRI score compared with the reference standard (ie, histology or 2-year follow-up). Results: A total of 1340 women (mean [range] age, 49 [18-96] years) were enrolled. Of 1194 evaluable women, 1130 (94.6%) had a pelvic mass on MRI with a reference standard (surgery, 768 [67.9%]; 2-year follow-up, 362 [32.1%]). A total of 203 patients (18.0%) had at least 1 malignant adnexal or nonadnexal pelvic mass. No invasive cancer was assigned a score of 2. Positive likelihood ratios were 0.01 for score 2, 0.27 for score 3, 4.42 for score 4, and 38.81 for score 5. Area under the receiver operating characteristic curve was 0.961 (95% CI, 0.948-0.971) among experienced readers, with a sensitivity of 0.93 (95% CI, 0.89-0.96; 189 of 203 patients) and a specificity of 0.91 (95% CI, 0.89-0.93; 848 of 927 patients). There was good interrater agreement among both experienced and junior readers (κ = 0.784; 95% CI, 0.743-0824). Of 580 of 1130 women (51.3%) with a mass on MRI and no specific gynecological symptoms, 362 (62.4%) underwent surgery. Of them, 244 (67.4%) had benign lesions and a score of 3 or less. The MRI score correctly reclassified the mass origin as nonadnexal with a sensitivity of 0.99 (95% CI, 0.98-0.99; 1360 of 1372 patients) and a specificity of 0.78 (95% CI, 0.71-0.85; 102 of 130 patients). Conclusions and Relevance: In this study, the O-RADS MRI score was accurate when stratifying the risk of malignancy in adnexal masses.

Lycopene improves methotrexate-induced functional alterations of the Madin-Darby kidney cells in a concentration-dependent manner.

Lycopene is one of the most potent antioxidants among carotenoids due to its ability to quench singlet oxygen and react with free radicals to reduce DNA damage. Methotrexate is widely used in the treatment of several types of cancers and autoimmune diseases. One of the most common side effects of a high-dose of methotrexate is kidney injury. In this study, we evaluated effects of lycopene on the Madin-Darby canine kidney cells (MDCK) treated with methotrexate through the estimation of their mitochondrial and lysosomal functions ((4,5-dimethylthiazol-2-yl)-2,5-diphenyl-2H-tetrazolium bromide reduction assay and neutral red uptake assay) and changes in cell oxidative status (determination of advanced oxidized proteins concentrations and reduced glutathione levels) and lysosomal enzymes activity (ß-N-acetyl glucosaminidase activity). Results of our study showed that lycopene applied in high concentration caused significant impairment of the MDCK function leading to cell death. Contrarily, in relatively low concentrations lycopene moderately ameliorated methotrexate-induced MDCK cell death estimated by both biochemical and microscopic analyses. It also prevented a significant decline in the MDCK cell lysosomal function estimated by neutral red accumulation ability and activity of the lysosomal enzyme ß-N-acetyl glucosaminidase.

Systematic radiological approach to utero-ovarian pathologies.

Ultrasound is the first-line imaging modality for the evaluation of suspected adnexal masses, endometriosis and uterine tumors, whereas MRI is used as a secondary diagnostic tool to better characterize these lesions. The aim of this review is to summarize the latest advances in the imaging of these utero-ovarian pathologies.

The Effect of Conditioned Media of Stem Cells Derived from Lipoma and Adipose Tissue on Macrophages' Response and Wound Healing in Indirect Co-culture System In Vitro.

Immunomodulatory and wound healing activities of adipose-derived stem cells (ADSCs) have been reported in various in vitro and in vivo experimental models suggesting their beneficial role in regenerative medicine and treatments of inflammatory-related disorders. Lipoma-derived stem cells (LDSCs) were reported as a potential tool in regenerative medicine due to the similarity with ADSCs but we have previously shown that LDSCs have different differentiation capacity than ADSCs despite a similar mesenchymal phenotype. To further analyze the potential differences and/or similarities between those two stem cell types, in the present study we examined the macrophages (MΦs)' response, immunomodulatory and wound healing effect of conditioned media (CM) of LDSCs and ADSCs in indirect co-culture system in vitro. We confirmed similar mesenchymal phenotype and stemness state of LDSCs and ADSCs but indicated differences in expression of some inflammatory-related genes. Anti-inflammatory potential of CM of LDSCs and ADSCs, with pronounced effect of LDSCs, in unstimulated RAW 264.7 MΦs was evaluated by decrease in Tnf and increase in Il10 gene expression, which was confirmed by corresponding cytokines' secretion analysis. Conditioned media of both LDSCs and ADSCs led to the functional activation of MΦs, with slightly more pronounced effect of CM of LDSCs, while both stimulated wound healing in vitro in a similar manner. Results of this study suggest that LDSCs secrete soluble factors like ADSCs and therefore may have a potential for application in regenerative medicine, due to immunomodulatory and wound healing activity, and indicate that LDSCs through secretome may interact with other cells in lipoma tissue.

The Addition of High Doses of Hyaluronic Acid to a Biphasic Bone Substitute Decreases the Proinflammatory Tissue Response.

Biphasic bone substitutes (BBS) are currently well-established biomaterials. Through their constant development, even natural components like hyaluronic acid (HY) have been added to improve both their handling and also their regenerative properties. However, little knowledge exists regarding the consequences of the addition of HY to their biocompatibility and the inflammatory tissue reactions. Thus, the present study was conducted, aiming to analyze the influence of two different amounts of high molecular weight HY (HMWHY), combined with a BBS, on in vitro biocompatibility and in vivo tissue reaction. Established in vitro procedures, using L929 cells, were used for cytocompatibility analyses under the test conditions of DIN EN:ISO 10993-5. For the in vivo part of the study, calvarial defects were created in 20 Wistar rats and subsequently filled with BBS, and BBS combined with two different HMWHY amounts, i.e., BBS + HY(L) and BBS + HY(H). As controls, empty defects were used. Established histological, immunohistochemical, and histomorphometrical methods were applied to analyze the tissue reactions to the three different materials, including the induction of pro- and anti-inflammatory macrophages and multinucleated giant cells (BMGCs). The in vitro results showed that none of the materials or compositions caused biological damage to the L929 cells and can be considered to be non-toxic. The in vivo results showed that only the addition of high doses of HY to a biphasic bone substitute significantly decreases the occurrence of pro-inflammatory macrophages (* p < 0.05), comparable to the numbers found in the control group, while no significant differences within the three study groups for M2-macrophages nor BMGCs were detected. In conclusion, the addition of different amounts of HMWHY does not seem to affect the inflammation response to BBS, while improving the material handling properties.

Stem Cells Derived from Lipoma and Adipose Tissue-Similar Mesenchymal Phenotype but Different Differentiation Capacity Governed by Distinct Molecular Signature.

Lipomas are benign adipose tissue tumors of unknown etiology, which can vary in size, number, body localization and cell populations within the tissue. Lipoma-derived stem cells (LDSCs) are proposed as a potential tool in regenerative medicine and tissue engineering due to their similar characteristics with adipose-derived stem cells (ADSCs) reported so far. Our study is among the first giving detailed insights into the molecular signature and differences in the differentiation capacity of LDSCs in vitro compared to ADSCs. Mesenchymal stem cell phenotype was analyzed by gene expression and flow cytometric analysis of stem cell markers. Adipogenesis and osteogenesis were analyzed by microscopic analysis, cytochemical and immunocytochemical staining, gene and protein expression analyses. We showed that both LDSCs and ADSCs were mesenchymal stem cells with similar phenotype and stemness state but different molecular basis for potential differentiation. Adipogenesis-related genes expression pattern and presence of more mature adipocytes in ADSCs than in LDSCs after 21 days of adipogenic differentiation, indicated that differentiation capacity of LDSCs was significantly lower compared to ADSCs. Analysis of osteogenesis-related markers after 16 days of osteogenic differentiation revealed that both types of cells had characteristic osteoblast-like phenotype, but were at different stages of osteogenesis. Differences observed between LDSCs and ADSCs are probably due to the distinct molecular signature and their commitment in the tissue that governs their different capacity and fate during adipogenic and osteogenic induction in vitro despite their similar mesenchymal phenotype.

In Vivo Analysis of the Biocompatibility and Macrophage Response of a Non-Resorbable PTFE Membrane for Guided Bone Regeneration.

The use of non-resorbable polytetrafluoroethylene (PTFE) membranes is indicated for the treatment of large, non-self-containing bone defects, or multi-walled defects in the case of vertical augmentations. However, less is known about the molecular basis of the foreign body response to PTFE membranes. In the present study, the inflammatory tissue responses to a novel high-density PTFE (dPTFE) barrier membrane have preclinically been evaluated using the subcutaneous implantation model in BALB/c mice by means of histopathological and histomorphometrical analysis methods and immunohistochemical detection of M1- and M2-macrophages. A collagen membrane was used as the control material. The results of the present study demonstrate that the tissue response to the dPTFE membrane involves inflammatory macrophages, but comparable cell numbers were also detected in the implant beds of the control collagen membrane, which is known to be biocompatible. Although these data indicate that the analyzed dPTFE membrane is not fully bioinert, but its biocompatibility is comparable to collagen-based membranes. Based on its optimal biocompatibility, the novel dPTFE barrier membrane may optimally support bone healing within the context of guided bone regeneration (GBR).

Survey highlights the need for specific interventions to reduce frequent conflicts between healthcare professionals providing paediatric end-of-life care.

AIMS: This study explored how paediatric healthcare professionals experienced and coped with end-of-life conflicts and identified how to improve coping strategies. METHODS: A questionnaire was distributed to all 2300 professionals at a paediatric university hospital, covering the frequency of end-of-life conflicts, participants, contributing factors, resolution strategies, outcomes and the usefulness of specific institutional coping strategies. RESULTS: Of the 946 professionals (41%) who responded, 466 had witnessed or participated in paediatric end-of-life discussions: 73% said these had led to conflict, more frequently between professionals (58%) than between professionals and parents (33%). Frequent factors included professionals' rotations, unprepared parents, emotional load, unrealistic parental expectations, differences in values and beliefs, parents' fear of hastening death, precipitated situations and uncertain prognosis. Discussions with patients and parents and between professionals were the most frequently used coping strategies. Conflicts were frequently resolved by the time of death. Professionals mainly supported designating one principal physician and nurse for each patient, two-step interdisciplinary meetings - between professionals then with parents - postdeath ethics meetings, bereavement follow-up protocols and early consultations with paediatric palliative care and clinical ethics services. CONCLUSION: End-of-life conflicts were frequent and predominantly occurred between healthcare professionals. Specific interventions could target most of the contributing factors.

Spasmolytic, antimicrobial and cytotoxic activities of 5-phenylpentyl isothiocyanate, a new glucosinolate autolysis product from horseradish (Armoracia rusticana P. Gaertn., B. Mey. & Scherb., Brassicaceae).

Detailed analyses of horseradish autolysates led to the identification of a new natural product, 5-phenylpentyl isothiocyanate (PhPeITC). The structural assignment was corroborated by synthesis, and the identity unequivocally established by spectral means. The occurrence of PhPeITC is the first direct proof of the existence of a 5-phenylpentyl glucosinolate in the aerial parts of this species as one of the possible "mustard oil" precursors. To verify its possible contribution to the horseradish functional food status, horseradish above- and underground autolysates, together with five ω-phenylalkyl isothiocyanates were tested for their spasmolytic, cytotoxic and antimicrobial activities. Specifically, the cytotoxic effect on Caco-2, HeLa (cancer) and MDCK (non-cancer) cell lines was established. Additionally, the five tested ITCs exerted significant spasmolytic activity (on rat distal colon), with PhPeITC being almost 100 times more potent than papaverine. A non-selective antimicrobial activity of all ITCs was revealed in the case of 6 bacterial and 2 fungal strains.