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Developing plant derived chemopreventive agents that can divert the carcinogenic process and inhibit tumour progression may greatly reduce serious health consequences of cancer. This study investigated the antiproliferative and apoptosis inducing ability of Rubus fairholmianus root acetone (RFRA) on MCF-7 cells. RFRA treatments showed an increase in the in vivo antioxidant levels such as superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase (GPx) and glutathione (GSH) in mice. The extract showed significant antitumour activity with 76.57% increase in life span of ascites tumour bearing mice and 1.94 cm³ reductions in solid tumour volume at 250 mg/kg. The cytotoxicity spectrum analysis indicated that RFRA is a potent cytocidal agent with varying levels of toxicity (IC₅₀: 57.14-29.36 μg/ml). A significant dose dependent decrease in viability, proliferation and increase in cytotoxicity, caspase 3/7 activity were observed in MCF-7 cells. RFRA (20 μg/ml) induced apoptosis in MCF-7 cells, which was observed by morphology; DNA ladder formation and increased apoptotic population in flow cytometry analysis. These findings strongly suggest the use of R. fairholmianus as natural antioxidant with profound antitumour activities. It can act as a potent cytotoxic agent, which is able to induce apoptosis in MCF-7 cells via caspase 3/7 upregulation.

Detonation nanodiamonds (DNDs) are usually small particles of 4-5 nm, but in aqueous suspension, DNDs form agglomerates in sizes larger than 1 μm. We propose the use of Bead Assisted Sonic Disintegration and a carboxylation procedure, to reduce DNDs aggregates sizes to approximately 100 nm. High cost zirconium beads have been substituted by silica beads synthetized in our laboratory and less-time consuming conditions were standardized. Techniques as Dynamic Light Scattering (DLS), Fourier Transform InfraRed Spectroscopy (FTIR), Transmission Electron Microscopy (TEM) and X-ray Photoelectron Spectroscopy (XPS), have been used to characterize the resulting diamond nanoparticles. While the incubation of Red Blood Cells with partially disaggregated DNDs was used to study whether these nanodiamonds impact in a living system. Our results show the absence of a negative effect in cell viability as well as no differences between Raman spectra of hemoglobin (Hb), from control and cell + DNDs conditions.

Amyloidosis is a group of diseases caused by the accumulation of insoluble protein aggregates in different parts of the body. Repeated subcutaneous injection of insulin hormones in diabetic patients leads to localized amyloidosis that is found to be cytotoxic. Thus, agents that can dissociate these aggregates are critically needed. In the present study, insulin amyloid dissociation was demonstrated by the treatment of an enzyme lumbrokinase (LK) isolated from earthworm. Thioflavin T (ThT) fluorescence, solution turbidity, particle size analysis, FTIR, CD, atomic force microscopy and cell viability assay were employed to support the dissociation of insulin amyloid in vitro. The small animal optical imaging was used to explore the dissociation of amyloid fibrils in vivo using zebrafish model. The activity of LK towards amyloid dissociation was compared with the standard amyloid fibril degrading agent nattokinase (NK). Our results indicated that LK can be a probable fibril degrading agent for the dissociation of amyloids.

In the present investigation Hibiscus rosa-sinensis (petals), Acorus calamus (rhizome), Moringa oleifera (leaves) and Cucurbita maxima (petals) were screened for their efficacy against osteosarcoma with elucidating a mechanism of their anticancer potentiality. The methanolic plant extracts revealed the presence of all major phytochemicals with quantitative analysis of flavonoids (98.15 ± 2.02 to 12.34 ± 0.57 mg of RUE/g) and total phenolics (26.40 ± 0.11 to 8.54 ± 0.10 mg of GAE/g). The antioxidant activity was assessed by standard DPPH, H₂O₂ scavenging, NO scavenging assays. The hemolysis, hemagglutination, and erythrocyte aggregation assays unveiled their compatibility with blood components. As most of the opportunistic microbes infect subsequently immunocompromised patients, the antimicrobial activity of the plant extracts showed a zone of inhibition (in mm) against nosocomial strains of Bacillus subtilis, Escherichia coli, Pseudomonas aeruginosa and Vibrio cholerae having MIC between 12.5-50 μg/ml. Through MTT assay the IC₅₀ value was calculated against MG 63 osteosarcoma with detailed studies on DNA fragmentation and chromatin condensation, revealing apoptosis being their primary mode of anticancer effect. Further the migration and colony forming assays supported the anticancer potentials of the methanolic plant extracts. The cell cycle analysis revealed that A. calamus and M. oleifera extracts were capable of arresting the growth of MG 63 osteosarcoma cells.

This study investigated the FSH influence on maturation rates of oocytes in vitro maturation (IVM), and expression levels of the follicle-stimulating hormone receptor (FSHR), luteinizing hormone receptor (LHR) and gonadotropin releasing hormone receptor (GnRHR) of cumulus-oocyte complexes (COCs) response to FSH treatment. 1686 COCs were harvested from 1063 ovaries of sheep. COCs were cultured 26 h at 38.5 °C and 5.0% CO₂ in IVM media supplemented with 0, 5, 10, 20 and 30 IU/ml FSH. They were allocated in to FSH-1 (basal line), FSH-2, FSH-3, FSH-4 and FSH-5 groups. The apoptosis of COCs was assessed by Tunel assay. Expression levels of mRNA and protein for FSHR, LHR and GnRHR in sheep COCs were detected using real time RT-PCR and Western blotting respectively. The results showed that the maturation rates of oocytes were improved gradually when FSH supplement increased from 0 to 10 μg/ml. FSH-3 group showed the highest maturation rate. Apoptosis rates of FSH-treated groups were less than that of FSH-1 group with a minimum of FSH-3 group. Expression levels of FSHR and LHR mRNAs in FSH-3 and FSH-4 were significantly higher than in FSH-1. Expression level of GnRHR mRNA in FSH-3 was higher than in FSH-1 (P < 0.05). Expression levels of FSHR proteins in FSH-3 and FSH-4 groups were higher than that of FSH-1 group. Expression levels of GnRHR proteins increased gradually with a maximal increment of FSH-5. Maturation rates of COCs had significant positive correlations with mRNA and protein levels of FSHR, LHR and GnRHR. In conclusion, FSH could accelerate the maturation rate of sheep oocytes and reduce their apoptosis rate, also increase the expression levels of FSHR, LHR and GnRHR mRNAs, and strengthen expressions of FSHR and GnRHR proteins. 10 IU/ml FSH additions were the optimal dose for IVM of sheep oocytes.

Novel benzoylhydrazones were designed and synthesized by condensation of 5-methoxysalicylaldehyde and benzhydrazides with different substituents at 4th position. The structures of the new derivatives were confirmed by elemental and thermal analysis, mass, IR, ¹H NMR and ¹³C NMR spectroscopy. The molecular properties of the compounds, important for drug pharmacokinetics and biodisposition in the human body, were assessed by the Lipinski's rule of five. In silico evaluation of the LogP value and the remaining parameters of drug similarity, as well as the topological polar surface area and absorption percentage, were used only as a first step in the study. The investigated 5-methoxyderivative hydrazones were further tested for in vitro cytotoxicity on three leukemic, two breast cancer and one non-tumor human cell lines using the MTT-dye reduction assay. The bioassay demonstrated that the compounds exhibited concentration-dependent antiproliferative activity at low micromolar concentrations against the used human cell lines. The solid tumor-derived breast cancer cell lines were generally more sensitive to the effects of the hydrazones with IC₅₀ values ranging 0.91 μM-12.07 μM. The results confirm that all compounds are more potent than the standard drug Melphalan and have appropriate properties as potential anti-breast cancer drug candidates.

Candida, a commensal and opportunistic fungal pathogen has been typically known for its biofilm forming ability and device-associated hospital acquired infections in human. The study aimed at exploring the in vitro anti-biofilm and anti-metabolic activity of AgNPs against C. albicans (n = 2),C. tropicalis (n = 2) and C. parapsilosis (n = 2) isolated from urine samples. Broth dilution method revealed greater than 50% inhibition at 100 ppm against Ag NPs in 24 h. An overall reduction of 55-86% in biomass (crystal violet staining assay) and 20-73% in metabolic activity (XTT assay) was observed in 24 h old biofilms. However, C. albicans proved to be more susceptible to AgNPs compared to C. tropicalis and C. parapsilosis. Scanning Electron Microscopy revealed patchy growth and deterrence in biofilm biomass when Ag NPs were coated on urinary catheter. Furthermore, viable cell counts of Candida were significantly reduced on AgNPs coated catheter compared to control.

The migration potency of differentiated (H-4-II-E, H-4-II-E-C3) and dedifferentiated (H-5) cell lines originated from Reuber hepatoma H-35, and their connection to the morphology and expression of gangliosides were examined. The migration capacity tested by a Transwell assay was fifteen times higher in H-5 cells compared to H-4-II-E cells. The ganglioside pattern was assessed by thin-layer chromatography (TLC). H-4-II-E and H-4-II-E-C3 cell lines expressed Fuc-GM1, which was not found in H-5 cells, whereas H-5 expressed GM3, which was absent in differentiated cell lines. GM3 ganglioside is thought to be one of the key molecules involved in signal transduction of mammalian cells. We conclude that changes in the migration capacity of various hepatoma cell lines might relate to their ganglioside spectra.

Ionising radiation (IR) is one of the main treatment modalities in oncology. However, we still search for substances which can radio-sensitize tumour cells. In this study we used caffeine, a non-specific ataxia-telangiectasia mutated kinase (ATM) inhibitor, and studied its effect on the activation of the proteins involved in cell cycle control and the induction of apoptosis in human T-lymphocyte leukaemic MOLT-4 cells (p53 wt). We evaluated the expression of the tumour-suppressor p53 (itself and phosphorylated on Ser¹⁵ and Ser³⁹²), the cell cycle regulator p21, and the anti-apoptotic protein myeloid cell leukemia 1 (Mcl-1). After treatment with 2 mM caffeine, the cells were irradiated by 1 or 3 Gy, lysed and the proteins detected by Western-blotting. Apoptosis was determined by flow-cytometric annexin V/propidium iodine detection. Irradiation by 1 or 3 Gy induced p53 phosphorylation at Ser¹⁵ and Ser³⁹² after 2 h with maximum after 4 h. Adding caffeine significantly inhibited Ser¹⁵ phosphorylation, which is ATM-dependent but surprisingly also Ser³⁹² phosphorylation, which is ATM-independent, suggesting that caffeine might have another cellular target (protein kinase). Similarly, caffeine caused a substantial decrease in p21 in combination with both doses of IR and also Mcl-1 was down-regulated. Three days after irradiation, caffeine significantly increased induction of apoptosis. The ATM/p53 pathway was suppressed by caffeine, which led to increased apoptosis accompanied by a p53-independent decrease in Mcl-1. It also caused down-regulation of p21, which possibly contributed to the shortened cell cycle arrest necessary for effective DNA repair and thus impeded radio-resistance. Caffeine promotes the cytotoxic effect of ionising radiation and provides a possible platform for the development of new anti-cancer therapeutics known as radio-sensitizers.

Recent years have seen mounting evidence for the role of melatonin in mediating programmed cell death, with a protective, anti-apoptotic effect in healthy cells, but an anti-tumoural, pro-apoptotic action in many tumour cells. In this study, we evaluated the effect of melatonin on the programmed cell death induced by thapsigargin (TG), on lymphocytes and neutrophils, and on various tissues obtained from rats injected with human promyelocytic leukæmia cells (HL-60), treated with melatonin in their drinking water (20 μm), and fed ad libitum. Melatonin treatment significantly reduced caspase-3 and -9 activity, and caused the proportions of lymphocytes, neutrophils, and eosinophils to revert to their basal values. No histological differences were observed. In conclusion, melatonin has anti-apoptotic effects on lymphocytes and neutrophils obtained from rats injected with HL-60 leukæmia cells.

The present study aimed to investigate the roles of fibrin deposition and protease activated receptor-1 (PAR-1) in renal cytokine/chemokine production and inflammatory cell infiltration in rats of different ages. Acute inflammation was induced by lipopolysaccharide (LPS) in rats which were then treated with tranexamic acid (TA), TA+urokinase (UK) or TA+low-molecular-weight heparin (HP). Fibrin deposition, inflammatory cells and expressions of PAR-1, monocyte chemoattractant protein-1 (MCP-1) and intercellular adhesion molecule 1 (ICAM-1) were detected. A reduction in fibrin deposition and PAR-1 expression in the LPS+TA+HP group was associated with decreased infiltration of inflammatory cells and down-regulated expressions of MCP-1 and ICAM-1. In the LPS+TA+UK group, the fibrin deposition, but not the PAR-1 expression, was reduced, However, the infiltration of inflammatory cells decreased and the expressions of MCP-1 and ICAM-1 down-regulated. There were significant differences in the fibrin deposition, infiltration of inflammatory cells and expression of PAR-1, MCP-1 and ICAM-1 between young and old rats undergoing the same treatment. These findings demonstrated that fibrin deposition plays more important roles than PAR-1 dose in cytokine/chemokine production and inflammatory cell infiltration in vivo, and ageing may deteriorate the fibrin deposition-induced production of cytokines/chemokines and infiltration of inflammatory cells.

This study aimed to investigate the effect of phenylhydrazine-induced hemolytic anemia on testicular functions and protective role of crocin in mice. Forty-nine adult male mice were studied in 7 groups. The control mice received normal saline, three groups were treated with 2, 4, and 6 mg/100 g of phenylhydrazine, and three other groups received 20 mg/100 g of crocin with phenylhydrazine for 35 days. Then, the blood samples were taken to examine oxidative stress of serum, sperm samples were obtained for IVF testing, and testicle tissue samples were taken for morphological studies. Morphometric results indicated a significant reduction in TDI (tubular differentiation index), RI (repopulation index or number of type B spermatogonia), and SI (spermiogenesis index) factors, number of Sertoli and Leydig cells, and diameter of germinal epithelium in the groups receiving phenylhydrazine. Histochemical results indicated some changes in the metabolic cycle of the testicle and results of serum tests showed variations in the peroxidation of lipids and antioxidant capacity of serum. Also, hemolytic anemia significantly reduced testicular parameters and crocin minimized the resulting injuries. It can be concluded that crocin is able to neutralize the complications which are resulting from the hemolytic anemia relating to testicular parameters.

Human amniotic epithelial cells (hAEC) are characterized by a great ability to differentiate, and immunomodulatory properties toward mother's immune system cells. These features have been described as being able to change during pregnancy. Thanks to their unique properties: low immunogenicity and high effectiveness of transplantations, amniotic epithelial cells constitute a very attractive source of stem cells for practical purposes in regenerative medicine and transplantology. In this review, we focus on natural factors potentially determining hAEC immunophenotype during pregnancy. Recognition of the impact of specific factors on hAEC would help in effective isolation, creation of appropriate culture conditions and regulation of desired cell function. We also indicate immunomodulatory properties of hAEC themselves. Discovering relations of hAEC with the microenvironment seems to be crucial for their clinical application.

Autism spectrum disorders (ASD) are characterized by impairments in social interaction and communication along with stereotyped patterns of behaviors. Over the past several decades, the prevalence of ASD has increased dramatically. ASD are highly multifactorial, with many risk factors acting together. Our review suggests that most risk factors are connected to immunoexcitotoxicity. Fluoride exposure is common as a result of the artificial fluoridation of drinking water and a dramatic increase in the volume of man-made industrial fluoride compounds released into the environment. Human exposure to environmental aluminum is extensive and appears to be growing. The long-term fluoride and aluminum burden have several health effects with a striking resemblance to the ASD. Moreover, both fluoride and aluminum interfere with a number of glycolytic enzymes, resulting in a significant suppression of cellular energy production. The synergistic interactions of fluoride and aluminum increase the potential neurotoxic effect particularly in children. Aluminofluoride complexes have effects on cell signaling, neurodevelopment, and neuronal function. We suggest that the burden with these new ecotoxicological factors could contribute to an alarming increase in the prevalence of ASD.

Biogenic metal nanoparticles owing to elimination of hazardous chemicals are beneficial for human healthy and biomedical applications. Considerable properties of these particles have motivated researchers to develop of the biosynthesized nanomaterials. In this study, the gold nanoparticles using Dracocephalum kotschyi leaf extract (d-GNPs) were synthesized and characterized by TEM-SEAD, SEM-EDAX, XRD, Zeta potential, DLS, and FT-IR analysis. TEM photograph showed spherical morphologies with an average size of 11 nm. SEAD pattern authorized fcc structure of these particles. The average zeta value of -29.3 mV revealed the surface charge of green synthesized GNPs. We studied the influence of different parameters such as reaction temperature, contact time, and pH on the synthesis of d-GNPs by UV-vis spectroscopy. Moreover, the obtained nanoparticles were evaluated regarding their biological activity (anti-cholinesterase and anticancer), as well as their influence on both the Gram classes of bacteria. The AuNPs showed an excellent inhibitory efficacy against AChE and BChE. Biological results exhibited that these particles displayed a dose-dependent cytotoxicity with IC₅₀: 196.32 and 152.16 μg/ml against K562 and HeLa cell lines as well. These metal nanoparticles were found to show no activity toward tested bacteria.

Betahistine dihydrochloride, which is widely prescribed for the treatment of symptoms associated with Meniere's syndrome, is generally administered orally in solid or liquid formulations. There is a strong need of profound investigation of alternative routes of administration of betahistine to overcome difficulties related to oral administration. The aim of this study was to evaluate betahistine cytotoxicity and permeability in vitro and to assess the drug's relevance for incorporation in drug delivery systems for nasal administration. RPMI epithelial model was used to evaluate drug permeability in vitro. The cytotoxicity of betahistine was assessed by MTT test. Chitosan microspheres were used as a betahistine delivery system. RPMI 2650 formed a thick, impermeable cell layer on the apical side of the filter inserts and developed enough TEER values to confirm confluence. According to the obtained results, BET showed high permeability coefficients (Papp values in the range 2.3 × 10^-5 to 19 × 10^-5) and could, therefore, be successfully used in nasal drug delivery formulations. Also, BET exhibited a good safety profile regarding nasal epithelium toxicity. A dose-dependent reduction in cell viability was observed. The microspheres as drug delivery systems affected BET permeation profiles due to the presence of chitosan as an absorption enhancer.

The earliest scientific journals on biomedicine began publication in the 50s and their authors addressed the application of biology to medicine. More recently, biochemistry and biomedical engineering questions have figured more prominently. This trend is discussed in a survey of the topics appearing in the Journal of Applied Biomedicine. Pharmacological and toxicological articles have been popular over the long term and the neurosciences, chronomedicine, molecular and cell biomedicine have also been very important. The role of computational biomedicine and nanomedicine has received increasing attention as has the part which applied biomedicine can play in the enhancement of the general economy.

Bone marrow mensenchyme cells (BMSCs) can differentiate into endothelial progenitor cells which then migrate to injured sites for the repair of neointima, and stromal cell-derived factor-1α (SDF-1α) can mediate the migration of CXCR4 expressing stem/progenitor cells to injured sites for pair. Protein and mRNA expression of SDF-1α and CXCR4 were determined by RT-PCR, Western blot and ELISA. Immediately after common carotid artery balloon injury, the mRNA expression of SDF-1α in vascular smooth muscle cells (VSMCs) first increased and then decreased 7 days later. VSMCs transfected with SDF-1α siRNA did not express SDF-1α mRNA, but after transfection with SDF-1α siRNA, the SDF-1α content in injured VSMCs gradually returned to the baseline level. Normal BMSCs rarely expressed CXCR4 mRNA, but the CXCR4 mRNA expression on BMSCs increased significantly 4 days after common carotid artery injury and was maintained. The migration of BMSCs after artery injury was enhanced when compared with normal BMSCs, but SDF-1α siRNA transfection of VSMCs and AMD3100 treatment remarkably decreased the chemotaxis of BMSCs to VSMCs and SDF-1α, respectively. We conclude that the SDF-1α/CXCR4 axis plays an important role in the migration of BMSCs after balloon injury and can ultimately cause abnormal proliferation of the intima.

There is a renewed interest in metabolism alterations and its impact on cancer development and progression. The metabolism of cancer cells is reprogrammed in order to support their rapid proliferation. Elevated fatty acid synthesis is one of the most important aberrations in cancer cell metabolism, and is required both for carcinogenesis and cancer cell survival. We have previously shown that cancer cells explore metabolic pathways especially autophagy and particularly enhanced glycolysis and suppressed oxidative phosphorylation to promote treatment resistance. To support cell proliferation in cancer, lipid metabolism and biosynthetic activities is required and often up-regulated. Here we bring lipid metabolic pathways into focus and summarized details that suggest a new perspective for improving chemotherapeutic responses in cancer treatment, and indicate the need to design more inclusive molecular targeting for a better treatment response.

Current biomedical research requires quantitative and qualitative studies of tissue metabolites. In this review, we discuss recent advances in multinuclear magnetic resonance (MR) technology that enhance biomedical research. The advances presented herein clearly show that MR is a diagnostic tool for the study of tumor tissue metabolism associated with changes in small molecule concentrations. Biomedical MR offers a non-invasive view of cancer cell metabolism by detection of resonance nuclei such as hydrogen-1, carbon-13, phosphorus-31 and fluorine-19. Due to current progress in MR technology, it is now possible to monitor changes in cancer cell metabolism before and after therapy.
Our criteria for the selection of research papers for this review were focused on those that show progress in biomedicine achieved by using MR. Our review demonstrates that small molecule detection in cancer tissue by MR has advanced biomedical research allowing for significant improvements in tumor detection and treatment.

The present work aimed at purifying the intracellular fungal metabolites, such as 16-methylheptadecanoic acid methyl ester (HDA) and 9,12-octadecadienoic acid (ODA) from marine Trichoderma, Hypocrea lixii TSK8, Hypocrea rufa SKS2 respectively, and investigating their anticancer and antioxidant effects. The two fungal metabolites were tested against two human cancer cell lines, namely oral cancer (KB) and skin carcinoma (A431) by using MTT assay. The inhibitory concentrations (IC₅₀) against KB oral cancer cells were found to be 18.75 ± 0.12 μg/mL for HDA and 75.50 ± 0.42 μg/mL for ODA. Whereas IC₅₀ values of HDA and ODA against A431 were found 37.5 ± 0.42 μg/mL and 72.89 ± 0.15 μg/mL, respectively. In addition, the down-regulation of heat shock protein 90 kDa (HSP90) was confirmed by using SDS-PAGE and Western blot analysis. The effect of HDA induced apoptosis via ROS-dependent internucleosomal DNA fragmentation was confirmed by AGE analysis. We further evaluated the in vivo anti-skin cancer activity of HDA in Swiss albino mice induced with skin cancer by 7,12-dimethylbenz(a)anthracene (DMBA) and croton oil (CO). The in vivo hematological, biochemical and histopathological results revealed that the fungal metabolite HDA was a highly potent anticancer compound against the skin cancer.

The integrity of whole blood samples may be compromised by suboptimal collection practices. Therefore, we investigated the influence of spurious hyperglycemia on erythrocytes and platelets, assessed with two hematological analyzers using optical or impedance technique. Three K₂EDTA blood specimens were collected from 12 healthy subjects, pooled and divided into four aliquots. The first aliquot was left untreated (glucose concentration 4.4 mmol/L), whereas scalar amounts of standard 5% glucose solution were added to the remaining, generating glucose contamination of 5% (19.2 mmol/L), 10% (33 mmol/L) and 20% (62 mmol/L). Hematological testing was then performed using Siemens Advia 2120 and Sysmex XE-2100, whereas glucose and cell-free hemoglobin were measured in centrifuged plasma. Hemolysis did not occur in contaminated aliquots, whereas hemoglobin, erythrocyte count, hematocrit, mean corpuscular hemoglobin concentration and platelet count progressively decreased with glucose concentration. No clinically significant variation was found for red blood cell distribution width and mean corpuscular hemoglobin. The erythrocyte size increased in parallel with glucose, exhibiting larger variation with Advia 2120 than with XE-2100. The mean platelet volume increased on Advia 2120 but not on XE-2100. These results suggest that hyperglycemia induces alterations of erythrocyte and platelet properties, which differ according to the technology used for assessment.

MSCs produce CD4(+)CD25(+)FOX3(+) regulatory T (Treg) cells from activated peripheral blood mononuclear cells (PBMC), T-CD4+ and T-CD8+ cells in vitro and in vivo. Here we investigated whether the deficiency of androgen/AR in MSCs influence Treg induction from total PBMC, splenocytes, CD4+CD25-through AR/TGF-β interaction. Eight to 12-week-old wild type and general androgen receptor knockout (ARKO) mice were used. MSCs were collected, characterized and function of Treg cells was studied. Our result showed that depletion of AR suppressed the immunosuppressive effect of MSCs, and demonstrated that WT-MSC-induced Treg cell expansion was partially impaired by blocking androgen receptor signal. Furthermore, the levels of TGF-β were lower in the T cell coculture with ARKO-MSC compared to WT-MSC. Exposure of ARKO-MSC cells to exogenous active TGF-β partially restored the induction of Treg cell expansion by ARKO-MSC cells. Our data suggest that ARKO-MSC hampers Treg cell expansion and function via androgen/AR and TGF-β signal pathways interaction. To the best of our knowledge, this study is the first investigating the interaction of MSCs from ARKO mice and WT Tregs in an allogeneic co-culture model. Together, these results might provide great insight into treatment of inflammatory and autoimmune diseases.

Cell proliferation is a crucial cellular process which influences development. In plants, meristems are formed by actively proliferating cells, in which the main expression of proliferation is the existence of a cell division cycle. Many cell activities are influenced by the cell proliferation status and cell cycle progression, among them ribosome biogenesis, which is morphologically expressed as the nucleolus. The connection is established through nucleolar proteins, which regulate the synthesis and processing of preribosomal precursors and, at the same time, are targets of various cell cycle regulators, such as certain kinases. Nucleolin is one of these nucleolar proteins, whose level increases with cell proliferation and depends on the cell cycle stages. Not only the levels, but also other important features of the protein, such as its distribution in situ in the nucleolus, its phosphorylation and its physiological degradation, depend on these parameters. Furthermore, since the nucleolar structure is highly sensitive to functional variations, distinct nucleolar structures, regarding the nucleolar size and the distribution of nucleolar subcomponents, have been defined for each period of the cell cycle, using synchronized cells. In addition to increasing our knowledge of cellular physiology, these relationships can be used to mark the proliferative state of the cell and the periods of cell cycle.

Streptococcus pneumoniae is the main cause of community acquired pneumonia and also produces meningitis, bacteremia, and otitis media, among others. Worldwide, these infections are the cause of substantial morbidity and mortality. Many different virulence factors have been described and most of them are surface-located macromolecules, namely, the capsular polysaccharide and various pneumococcal proteins. Cell wall hydrolases (CWHs) specifically cleave covalent bonds of the peptidoglycan and associated polymers: most CWHs are choline-binding proteins (CBPs) and are among the most well-known surface proteins. Pneumococcal CBPs have been investigated due to their role in pathogenesis and as candidate antigens for improved vaccines. Among the complex host-parasite interactions characteristic of pneumococcal disease, nasopharyngeal colonization is the first step. CBPs appear to play a central role in the development of the carrier state, possibly by affecting biofilm formation and development. Although the role of biofilms in the pathogenesis of some chronic human infections is currently widely accepted, the molecular bases underlying the formation of pneumococcal biofilms are only recently being studied. Among therapeutic strategies to combat multidrug-resistant pneumococcal infections, the use of purified phage- or bacteria-encoded CWHs both in vitro and in animal models is under investigation.

Nucleolin is the major nucleolar protein of animal, plant and yeast proliferating cells. It is enriched in the most soluble nuclear or nucleolar protein extract, containing ribonucleoproteins, from which it has been purified. It has a tripartite structure in which each domain accounts for different functions. Despite its multifunctionality, the best characterized role of nucleolin is in the primary cleavage of pre-rRNA, an early step of ribosome biogenesis. In the nucleolus of proliferating cells, nucleolin is mostly located in the dense fibrillar component, following a vectorial pattern, from the periphery of fibrillar centers outwards. This pattern is lost in quiescent cells in which nucleolin is present in low levels. Nucleolin is the most phosphorylated protein of the soluble nuclear extract. It is phosphorylated by casein kinase II and CDKA, and phosphorylation is closely associated with the role of nucleolin in proliferating cells. During mitosis, nucleolin is transported from the mother to the daughter cell nucleolus in the form of processing particles, together with pre-rRNA precursors and other nucleolar proteins. It forms part of prenucleolar bodies and plays a role in nucleologenesis. Recent studies on the nucleolin function, carried out on samples with inactivated nucleolin genes (siRNA downregulated or mutants) have evidenced that nucleolin is absolutely essential for cell proliferation, for the organization of the nucleolus and for transcription and processing of pre-rRNA. In plants, nucleolin controls the auxin responsiveness, thus being involved in the regulation of plant development.

The lipid components of pathogen cell membranes have been considered as a poor pharmacological target, due to their universal distribution and apparent homogeneity throughout living organisms. Among the rare exceptions to this view one could mention polyene antibiotics such as amphotericin, or peptide antibiotics such as the polymyxins and the gramicidins. In the last two decades, however, the above notion has been challenged by two main lines of discovery; first, natural antimicrobial peptides (AMPs) that kill pathogens by interaction with phospholipids and membrane permeabilization, and secondly, cell-penetrating peptides (CPPs), capable of introducing into cells a variety of cargoes in the absence of specific receptors, again by interaction at some point with membrane phospholipids. For both AMPs and CPPs, the pharmacological proof-of-concept has been successfully demonstrated, and promising applications as nanobiotechnological tools have been envisaged though not hitherto materialized in clinical settings. In this review we briefly examine the pros and cons of these two classes of therapeutic agents, as well as strategies aimed at rationalizing and expanding their potentiality.

Toxin-antitoxin systems (TAS) emerged more than 25 years ago and have since developed as an important field in molecular microbiology. TAS are autoregulated operons coding a stable toxin and an unstable antitoxin found in the plasmids and chromosomes of Bacteria and Archaea. The conditional activation of their toxins interferes with cell growth/viability and, depending on the context, can influence plasmid maintenance, stress management, bacterial persistence, cell differentiation and, it is likely also bacterial virulence. This review summarizes recent results on the parD system of plasmid R1 and on the chromosomal relBE systems found in Escherichia coli and in Streptococcus pneumoniae with a focus on the RNase activity of their toxins, their regulation and their biomedical applications and implications.

To resolve the problem of the insufficient availability of seed cells and to provide seed cells for tissue engineering research, an immortalized human bone marrow stromal stem cell line (MSCxj cells) was established in our department to investigate the ectopic osteogenesis of MSCxj cells.
MSCxjs were grown with a heterogeneous bone scaffold for 48 h. Three groups were included: group A: MSCxjs of 35 PDs were maintained with heterogeneous bone; group B: MSCxjs of 128 PDs were maintained with heterogeneous bone; and group C: heterogeneous bone alone. Tetracycline fluorescence staining, H&E staining, and ponceau staining, immunohistochemistry and bone histomorphometry were performed. At the same time, scanning electron microscopy was conducted to detect the growth of MSCxjs and heterogeneous bone.
Scanning electron microscopy showed favorable adherence of MSCxjs to heterogeneous bone. A large number of newly generated filamentous extracellular matrix and fine granular materials were found to cover the cells. The results from staining showed that the osteogenesis was not obvious in group A/B 4 weeks after transplantation. Eight weeks after implantation, osteoid matrix deposition was noted in and around the heterogeneous bone in group A/B. Twelve weeks after implantation, osteogenesis was increased in group A/B. There were no significant differences in the time course for bone formation and the amount of newly generated bone between group A/B.
Like primary hBMSCs, MSCxj cells have favourable ectopic osteogenesis and can be applied as seeded cells in bone tissue engineering.

Alpha-tomatine is a major glycoalkaloid found in the roots, leaves, stems and fruit of tomatoes Lycopersicon esculentum. Recently, alpha-tomatine has been recognized as a potential anticancer drug. In the present study, we identified the signaling cascades involved in the antitumor effect of alpha-tomatine on MOLT-4 leukemic cells. Alpha-tomatine inhibited the proliferation and decreased the viability of MOLT-4 cells in a dose-dependent manner. An increase in the activity of caspases 9 and 3/7 was not observed. However, an increase in the amount of p53 and its phosphorylation on serine 15, as well as an increased amount of mitochondrial protein PUMA was detected 4 and 24 h after exposure to alpha-tomatine at a concentration of 1-3 μmol/l. Inhibition of the proliferation of MOLT-4 cells by alpha-tomatine is also associated with an increase in p21^WAF1/CIP1 and the activation of Chk2. The comet assay did not detect significant amounts of single or double DNA strand breaks in cells treated with alpha-tomatine at concentrations of 0.1-9 μmol/l. Our results thus contribute to the understanding of the anticancer action of alpha-tomatine.