Showing papers in "in Vivo in 2015"

Curcumin suppression of cytokine release and cytokine storm. A potential therapy for patients with Ebola and other severe viral infections.

Abstract: Background: The terminal stage of Ebola and other viral diseases is often the onset of a cytokine storm, the massive overproduction of cytokines by the body's immune system. Materials and Methods: The actions of curcumin in suppressing cytokine release and cytokine storm are discussed. Results: Curcumin blocks cytokine release, most importantly the key pro-inflammatory cytokines, interleukin-1, interleukin-6 and tumor necrosis factor-α. The suppression of cytokine release by curcumin correlates with clinical improvement in experimental models of disease conditions where a cytokine storm plays a significant role in mortality. Conclusion: The use of curcumin should be investigated in patients with Ebola and cytokine storm. Intravenous formulations may allow achievement of therapeutic blood levels of curcumin.

Radical-scavenging and Anti-inflammatory Activity of Quercetin and Related Compounds and Their Combinations Against RAW264.7 Cells Stimulated with Porphyromonas gingivalis Fimbriae. Relationships between Anti-inflammatory Activity and Quantum Chemical Parameters.

Abstract: Background/aim The flavonoid quercetin exerts significant anti-inflammatory activity against chronic infections, including periodontal disease. However, it is unclear whether combination of quercetin with other flavonoids enhances antioxidant and anti-inflammatory activity. To clarify the molecular mechanism responsible for the anti-inflammatory activity of quercetin, we investigated the antioxidant, cytotoxicity and anti-inflammatory activity of quercetin and its related compounds, catechin and epicatechin, and their combinations. Materials and methods Radical-scavenging activities were determined by the 1,1-diphenyl-2-picrylhydrazyl (DPPH) assay, and cytotoxicity against RAW264.7 cells was determined using a cell counting kit (CCK-8). The inhibitory effects of these compounds on the mRNA expression of cyclooxygenase-2 (Cox2), tumor necrosis factor-alpha (Tnfα) and nitric oxide synthase 2 (Nos2), in RAW264.7 cells stimulated with Porphyromonas gingivalis (Pg) fimbriae, was also determined using real-time polymerase chain reaction analysis. The phenolic O-H bond dissociation enthalpy (BDE) and quantum chemical parameters were calculated on the basis of density function theory (DFT) BLYP/6-31G*. Results The DPPH(•) radical-scavenging activity (EC50) of quercetin, catechin and epicatechin was 5.5, 7.7 and 6.2 μM, respectively, whereas the cytotoxicity (LC50) was 4.45, 4.80 and 4.95 mM, respectively. Quercetin had slightly higher cytotoxicity and anti-DPPH(•) activity than catechin and epicatechin. The BDE for the three flavonoids at the 4'-OH in the B ring, which is the initial active site, was about 75 kcal/mol. Furthermore, various combinations of quercetin with catechin or epicatechin exerted an antagonistic effect on anti-DPPH(•) activity. Gene expression of Cox2, Tnfα and Nos2 stimulated by exposure to Pg-fimbriae was markedly suppressed by quercetin, but was not modulated by its combination with epicatechin. The 50% inhibitory concentration of quercetin for Cox2 expression was approximately 10 μM, while that of catechin and epicatechin was approximately 500 μM. Values of the quantum chemical parameters softness (σ) and electronegativity (χ) were highest for quercetin among the three flavonoids tested. Conclusion The potent anti-inflammatory activity of quercetin appears to be attributable to its high σ and χ values. Quercetin may be applicable as a preventive agent against inflammatory periodontal disease as a manifestation of systemic disease.

Review: The Prolonged QT Interval: Role of Pro-inflammatory Cytokines, Reactive Oxygen Species and the Ceramide and Sphingosine-1 Phosphate Pathways.

Abstract: Patients with QT prolongation have delayed cardiac repolarization and may suffer fatal ventricular arrhythmias. To determine the role of cytokines in causing this syndrome, we reviewed reports on patients with rheumatoid arthritis, psoriasis and other inflammatory conditions. These patients frequently have prolonged QT, which correlates with increases in tumor necrosis factor alpha, and interleukin-1β and 6. Studies in experimental models have shown that these cytokines act through stimulation of reactive oxygen species. Our review of data on phospholipidosis and on QT-shortening agents suggests a key role in QT prolongation for the ceramide/sphingosine-1-phosphate rheostat. We conclude that the cause of prolonged QT in inflammatory conditions is cytokine induction of reactive oxygen species and then ceramides, and believe that QT-prolonging agents bypass initial steps of this pathway and directly affect ceramides. Since both pro-inflammatory cytokines and numerous medications cause QT prolongation and ventricular arrhythmias by this mechanism, extra caution is needed when using these agents in patients with inflammatory conditions.

Octyl-2-cyanoacrylate Adhesive for Skin Closure: Eight Years Experience

Abstract: UNLABELLED Cyanoacrylate skin adhesive is increasingly used to ensure and stabilize wound closures. One of the documented favorable effects of these glues is to limit the risk of a surgical site infection by physically isolating the wound. PATIENTS AND METHODS From January 2005 to January 2013, 900 patients requiring surgical treatment in the Structure of Plastic Surgery of the University of Sienna were enrolled for the study and divided in two groups; 450 patients treated with glue versus 450 controls. RESULTS The cohort of patients treated with glue had lower incidence of wound infection and wound dehiscence. Approximately 375 work-hours were economized. CONCLUSION Octyl-2-cyanoacrylate is a valuable aid for ensuring the success of surgery by reducing surgical site infections. The use of octyl-2-cyanoacrylate is easy, guarantying a major superficial protection without relevant contraindications and with evident saving of time and resources.

Inhibition of Cell Growth of the Prostate Cancer Cell Model LNCaP by Cold Atmospheric Plasma.

Abstract: UNLABELLED Backround/Aim: Physical plasmas are ionized gases containing several biologically-reactive factors that yet exert their anti-microbial and anti-proliferative effects in fields of surface sterilisation, de-contamination and wound healing. MATERIALS AND METHODS Cold atmospheric plasma (CAP) was generated via the atmospheric pressure plasma jet kINPen09. Apoptotic effects of CAP treatment on the human epithelial prostate cancer cell line LNCaP as a cell culture model for malignant tumor tissue was analyzed by cell counting, western blot and quantitative reverse transcription-polymerase chain reaction analysis. RESULTS LNCaP cells exhibited significantly reduced cell growth following CAP treatment. We show that most probably the induction of apoptosis is the terminus of CAP treatment illustrated by the pro-apoptotic modulation of p53, p21, caspase-3, Bax, and survivin, as well as morphological changes of cell architecture. CONCLUSION Our in vitro study offers first indicatory results for molecular response mechanisms after CAP treatment in a suitable LNCaP cell model.

Enrichment and Schwann Cell Differentiation of Neural Crest-derived Dental Pulp Stem Cells.

Abstract: Background/aim As already described in previous studies, neural crest stem cells (NCSCs) can be found in adult human dental pulp. The present study investigated the methodology for enrichment and differentiation-induction of the above mentioned cells. Materials and methods Dental pulp was extracted from human wisdom teeth of four patients and subsequently cultured as explants on fibronectin-coated plates in neurobasal medium supplemented with B27, basic fibroblast growth factor (bFGF), epidermal growth factor (EGF), insulin, l-glutamine and neuregulin-β1. The cells were then characterized by immunofluorescence, while their differentiation-potential was tested by the attempt to induce cells into different lineages, i.e. osteogenic, melanocytic and glial. Results The enriched cell population expressed nestin, CD271 and SOX10, which are well-known markers for NCSCs. Consequently, the cells were successfully induced to differentiate into osteoblasts, melanocytes and Schwann cells, expressing the corresponding differentiation markers. Conclusion Human adult dental pulp contains a population of stem cells with neural crest ontogeny, which can thus be recruited for multiple regenerative therapies.

Assessment of Oxidative Stress in Septic and Obese Patients Using Markers of Oxidation-reduction Potential

Abstract: Background/aim The novel static (sORP) and capacity (cORP) oxidation-reduction potential markers were examined for assessing oxidative stress in plasma of patients with sepsis. Moreover, the possible effect of obesity-induced oxidative stress on patients with sepsis was investigated. Materials and methods sORP and cORP markers, as well as the conventional oxidative stress biomarkers total antioxidant capacity (TAC), thiobarbituric acid-reactive substances (TBARS) and protein carbonyls (CARB), were assessed in plasma. Results sORP marker was increased significantly in the sepsis group, while cORP was significantly lower compared to the control group, indicating oxidative stress. Furthermore, in patients with sepsis, TAC was significantly lower compared to control group. However, obesity had no effect on sORP, cORP and TAC in patients with sepsis, although it increased levels of CARB and TBARS. Conclusion The present results suggest, for the first time, that ORP markers could be used for assessing oxidative stress in patients with sepsis.

Abdominoplasty in Mildly Obese Patients (BMI 30-35 kg/m2): Metabolic, Biochemical and Complication Analysis at One Year.

Abstract: BACKGROUND/AIM Dermolipectomy dramatically improves the quality of life of previously obese patients. Fat removal in patients undergoing liposuction and reduction mammoplasty has positive effects, but no data regarding sequelae of abdominal dermolipectomy on metabolism have been reported. Thus, the aim of the present study was to assess metabolic and biochemical modifications occurring after abdominoplasty. A correlation between the results and the rate of complications was also evaluated. PATIENTS AND METHODS One hundred and twenty-eight patients with body mass index (BMI) between 30 and 35 kg/m(2) were enrolled. Clinical assessment, anthropometric measurements, blood biochemical analysis (fasting plasma glucose, total cholesterol, free fatty acids, plasma insulin), plasma adipocytokines (leptin, adiponectin,resistin), levels of inflammatory markers [tumor necrosis factor-alpha (TNFα), interleukin-6 (IL6), C-reactive protein (CRP)] and insulin sensitivity by Homeostasis Model Assessment were performed three times. RESULTS The average age was 37.6 years. At baseline, the mean (±SD) BMI of patients overall was 33.44 (±2.3) kg/m(2) and the amount of fat surgically removed was 1,578.33 (±1,013.02) g. A significant decrease in BMI at 1 year was found. Adiponectin levels were found to have significantly increased (p<0.05) in long-term evaluation. We noted significant increases in the mean TNFα (p<0.05), IL6 (p<0.01) and CRP (p<0.05), with a return to baseline values of all variables. The majority of patients were satisfied with the surgical procedure after 12 months. We noted a lower rate (10%) of complications in patients whose blood analysis showed an increase of adiponectin. CONCLUSION Abdominal dermolipectomy is a safe surgical procedure that improves the quality of life of previously obese patients from an aesthetic functional view. A statistically significant increase of adiponectin after the procedure was found. A low level of adiponectin after surgery indicates a higher probability of complications.

Silybin–Phosphatidylcholine Complex Protects Human Gastric and Liver Cells from Oxidative Stress

Abstract: Background/aim Silybin is the main component of silymarin with antioxidant, anti-inflammatory and cytoprotective actions. Our aim was to compare the effect of silybin used as single substance, silybin-phosphatidylcholine complex (SilPho), and derivatives of silybin (MannpSil, GalpSil, GlcpSil, LactpSil) on MKN28 and HepG2 cell viability and cell death, in vitro, after induction of oxidative stress. Materials and methods Oxidative stress was induced by incubating HepG2 and MKN28 cells with xanthine oxidase in the presence of its substrate xanthine. Cell viability was determined by the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazoliumbromide assay. Determination of Malondialdehyde (MDA) in MKN28 cells was performed by high-performance liquid chromatography. Quantitative analysis of apoptotic cells was carried-out using annexin. Results SilPho and new silybin glycoconjugates did not affect cell viability, while silybin induced about 50% cell death in both MKN28 and in HepG2 cells. Pre-treatment of cells with silybin and new silybin glycoconjugates (before oxidative stress induction) did not affect cell viability, while SilPho had a protective effect. Exposure of MKN28 cells to oxidative stress caused a two-fold increase in cellular MDA concentration compared to untreated cells. Moreover, pre-treatment with SilPho, but not with silybin, significantly prevented oxidative stress-induced increase in cellular Malondialdehyde. Moreover, silybin induced apoptosis potentiated by oxidative stress, while SilPho did not induce any effect. Oxidative stress caused cell death primarily by necrosis, antagonized by SilPho. Conclusion The protective effect of SilPho is partially due to inhibition of radical oxidative species.

β-Glucan from Saccharomyces cerevisiae Induces IFN-γ Production In Vivo in BALB/c Mice.

Abstract: Aim β-Glucan is one of the most abundant polymers in nature and has been established as an immunomodulator. This compound has notable physiological effects on mammalian immune systems, including anti-tumor and anti-infective activities and can activate the immune response. It is considered that the immune-stimulating activities of β-glucan can depend on physicochemical parameters, such as molecular size. Saccharomyces cerevisiae, also known as baker's yeast, is a frequently used source of β-glucan. The aim of the experiments was to investigate how different Saccharomyces cerevisiae β-glucan preparations with different molecular size affect interferon-gamma (IFN-γ) production in BALB/c mice. Materials and methods In vivo and in vitro BALB/c mouse models were used for the investigations. Different β-glucan preparations were orally administrated in the in vivo experiments. IFN-γ production in BALB/c mice was analyzed by enzyme-linked immunosorbent assay and measuring interferon-γ RNA concentration. Results The results showed that orally-administered β-glucan from S. cerevisiae enhanced IFN-γ production in BALB/c mice in the in vivo model, but not by mouse leukocytes in vitro. Moreover, water-soluble β-glucan enhanced IFN-γ production more effectively than did particulate β-glucan. Conclusion IFN-γ plays an important role in immunity against viral and bacterial infections. Our experiments have shown that β-glucan preparations enhance IFN-γ production in BALB/c mice and can be potentially used for immune system stimulation in mammals. Current results may be used to develop soluble β-glucan nutritional supplements.

Small leucine rich proteoglycans are differently distributed in normal and pathological endometrium.

Abstract: BACKGROUND During the woman's fertile period, the non-pregnant uterus is subject to constant cyclic changes. The complex mechanisms that control the balance among proliferation, differentiation, cell death and the structural remodeling of the extracellular matrix can contribute to the benign or malignant endometrial pathological state. The small leucine-rich proteoglycans (SLRPs) are important components of cell surface and extracellular matrices. MATERIALS AND METHODS Using immunohistochemistry, we showed that the distribution patterns of SLRPs were completely modified in the pathological compared to normal endometrium. RESULTS The expression of SLRPs was low/absent in all endometrial pathologies examined compared to normal endometrium. We observed an increase of lumican from proliferative to secretory phase of the endometrium and a decrease of fibromodulin, biglycan and decorin. In menopause endometrial tissue, the level of expression of fibromodulin, biglycan, decorin and lumican dramatically decreased. CONCLUSION The results revealed the prominence and importance of proteoglycans in the tissue architecture and extracellular matrix organization.

Review: The Role of Insulin-like Growth Factor-1 Signaling Pathways in Uterine Leiomyoma.

Abstract: A growing body of evidence suggests the association of the IGF-I bio-regulatory system with leiomyoma occurrence and growth. The complex interplay between IGF-I/IGF-IR and hormonal and other growth factors is, thus, now receiving significant attention. Elucidation of the molecular mechanisms driving the disease may allow for development of novel targeted-therapeutic strategies for the treatment of leiomyomas. Herein, we provide a concise update and overview of the function and regulation of IGF-I and its role in leiomyoma growth.

Outcomes of laparoscopic and open surgery for colorectal cancer in the emergency setting.

Abstract: Aim To evaluate the short-term and oncological outcomes of laparoscopy compared to open procedures for colorectal cancer in the emergency setting. Patients and methods Forty-nine consecutive patients undergoing emergency open (n=38) or laparoscopic (n=11) major resection were analyzed. Results There were no significant differences in age, gender, American Society of Anesthesiologists (ASA) score, body mass index (BMI), history of previous abdominal surgery and tumor location between the open and laparoscopy groups. Pathological T4 tumors were more commonly observed in the open-surgery group than the laparoscopy group (47% vs. 9%; p=0.022). Obstruction (45% vs. 27%) and perforation (42% vs. 27%) were also more common in the open-surgery group. Bleeding or anemia was more common in the laparoscopy group (8% vs. 45%; p=0.028). There were no open conversions in the laparoscopy group. Regarding the type of surgery, Hartmann's operation was more common in the open-surgery group (32%) and right hemicolectomy was more common in the laparoscopy group (36%; p=0.058). There was no difference in the 30-day complication rate of open surgery (34%) and laparoscopy (36%) (p=0.895). Mean time to tolerable diet (8 days vs. 6 days, p=0.035) and mean length of hospital stay (17 days vs. 13 days, p=0.041) were shorter in the laparoscopy group. Overall recurrence and cancer-specific death did not differ between the two groups. Conclusion In selected colorectal cancer patients, emergency laparoscopy confers benefits in terms of short-term and oncological outcomes. Therefore, experienced laparoscopic surgeons may more actively consider the use of laparoscopy in the emergency setting.

Polydatin administration improves serum biochemical parameters and oxidative stress markers during chronic alcoholism: a pilot study.

Abstract: AIM Polydatin, a hydroxystilbene derived from the rhizome of Polygonum cuspidatum, elicits hepatoprotective and neuroprotective effects through its anti-oxidant properties. The present study aimed to determine the effects of oral administration of polydatin in alcoholic patients in order to improve liver biochemical parameters, serum oxidative stress and mental state. We enrolled 20 chronic alcoholic patients hospitalized for rehabilitative therapy. The patients were divided into two groups receiving the following treatment regimes for two weeks: administration of an anti-oxidant nutritional supplement containing glutathione and vitamin C (group 1), or glutathione, vitamin C and polydatin (group 2). RESULTS The results of the present study show that elevated plasma aspartate aminotransferase and alanine aminotransferase levels in patients after two weeks of alcohol withdrawal were significantly reduced by polydatin (group 2), when compared to group 1. Polydatin also significantly reduced lipid peroxidation levels. Finally, our preliminary data resulting from the analysis of the Mini-Mental Status suggest that polydatin improves cognitive performance. CONCLUSION Daily dietary administration of polydatin should be considered for prevention and treatment of liver disease and cognitive impairment in alcoholic patients.

Ethanol Extract of Hedyotis diffusa Willd Affects Immune Responses in Normal Balb/c Mice In Vivo

Abstract: Numerous clinical anticancer drugs are obtained from natural plants and Hedyotis diffusa Willd (EEHDW) has been used as a major component in Traditional Chinese medicine formulas since a long time. Ethanol extracts of EEHDW have been shown to possess various biological activities including anticancer function in vitro. Our earlier studies have shown that EEHDW affects immune responses in WEHI-3-generated leukemia mice, but EEHDW has not been reported to affect immune responses in a normal mouse model. Herein, we investigated whether EEHDW could affect immune responses on normal murine cells in vivo. Normal BALB/c mice were orally treated with or without EEHDW at 0, 16, 32, and 64 mg/kg or 32 mg/kg by i.p. for 3 weeks, then were weighed, and blood, liver and spleen samples were collected for further experiments. Results indicated that EEHDW did not significantly affect body and liver weight but significantly increased the spleen weight by i.p. treatment when compared to control groups. Flow cytometric assays indicated that EEHDW promoted CD11b levels at 16, 32 and 64 mg/kg oral treatment, CD19 levels at 16, 32, 64 mg/kg oral treatment and i.p. treatment, and Mac-3 levels at 16, 32 and 64 mg/kg oral treatment, however, it did not significantly affect the levels of CD3. Oral treatment with 16 and 32 mg/kg of EEHDW significantly decreased macrophage phagocytosis from PBMC; 32 mg/kg of EEHDW by i.p. treatment significantly increased phagocytosis activity of macrophages obtain from the peritoneal cavity. EEHDW at 32 mg/kg by i.p. treatment led to an increase of NK cell activities compared to oil control groups. EEHDW at 32 mg/kg of EEHDW by i.p. treatment increased B- and T-cell proliferation. Based on these observations, EEHDW seems to have promoted immune responses in this murine model.

Mast Cells Stimulate Lymphangiogenesis in the Gingiva of Patients with Periodontal Disease

Abstract: The presence and distribution of lymphatic vessels and mast cells in the gingiva under normal and pathological conditions have been reported by several studies, but the relationship between them during inflammatory lymphangiogenesis is virtually unknown. The aim of the present study was to investigate the lymphatic microvessel density (LMVD) and mast cell density (MCD) in the gingiva of patients with periodontal disease compared to normal-like gingiva. Gingival punch biopsies from 51 patients with periodontal disease were investigated. MCs and LVs were detected by double-immunohistochemistry, using primary antibodies against mast cell tryptase and D2-40. The inflammatory infiltrate was evaluated on a scale from 0 (absent) to +3 (severe inflammation). MCs and LVs were counted in the same microscopic field for each case at ×200 magnification. We found a significant increase in the number of both MCs and LVs in cases with mild and moderate inflammatory changes, followed by a slight decrease in cases with severe inflammation. We have shown a particular association between MCs and LVs that may support the contribution of MCs to the development of the lymphatic vasculature in inflammatory conditions. MCD correlated with LMVD in all cases with mild and moderate inflammatory changes, but not in cases with severe inflammation. No correlation was found between MCD/LMVD and the density of the inflammatory infiltrate. Our results suggest the potential involvement of MCs in the induction and maintenance of lymphangiogenesis in the gingiva of patients with periodontal disease in the early steps of evolution.

Molecular Evidence of Apoptotic Pathway Activation in Semen Samples with High DNA Fragmentation

Abstract: Background/aim Male infertility is diagnosed by semen parameters, such as concentration, motility and morphology; however, these are not sufficient for the prediction of male fertility capacity. In the clinical routine, several other sperm functions have been introduced, including the sperm DNA fragmentation test. The objective of the present study was to evaluate sperm chromatin integrity in semen samples. Materials and methods Sperm chromatin dispersion test (SCD) was used in ejaculates from men divided into five groups: normozoospermic, oligozoospermic, asthenozoospermic, oligoasthenozoospermic and cryptozoospermic. Results The data obtained showed that the SCD percentage appeared to be significantly associated with oligozoospermia diagnosis. We also evaluated total testosterone, follicle-stimulating hormone (FSH), luteinizing hormone (LH) and inhibin B serum hormonal levels in all samples examined, in order to assess whether DNA fragmentation increase could correlate with abnormal hormonal values. Finally we selected certain samples with an increasing DNA fragmentation and analyzed the molecular activated apoptotic pathways. Conclusion A significant relationship was found between caspase-3 activation and increased DNA fragmentation.

The Nasopalatine Canal in Adults on Cone Beam Computed Tomograms-A Clinical Study and Review of the Literature.

Abstract: Aim The aims of this study were to assess the location, morphology and anatomical dimensions of the nasopalatine canal (NPC) on cone beam computed tomographic (CBCT) images and to compare our findings with recent reports on this matter. A detailed knowledge of anatomical variations is mandatory in skeletal surgery of the anterior maxilla. Patients and methods Two hundred CBCT scans of the mid-facial region were analyzed from adult patients at the Outpatient Clinic of the Department of Oral and Craniomaxillofacial Surgery, University of Hamburg, Germany. Patients were scanned using standard exposure time at normal patient positioning inside the CBCT device. Three-dimensional (3D) CBCT images were carefully analyzed regarding NPC location, morphology and anatomical dimensions, and variations of radiological morphology, with respect to age and gender. Results In the sagittal plane, the NPC typically appeared as a canal with a mean length±standard deviation (SD) of 11.15±2.87 mm. The oral cavity opening of the canal is the incisive foramen, with a mean diameter of 4.49±1.71 mm. At the entrance to the nasal floor, in most cases, two apertures were found (Y-canal morphology), but also three or four openings were observed. In particular cases, the canal presented a cylindrical aperture with only one exit to the nasal floor. The average width of the NPC at the level of the nasal floor was 3.43±1.54 mm. The labiopalatal width of the NPC measured perpendicular to the long axis of the canal on sagittal plane was 2.48±1.33 mm. Interpretation of NPC morphology was significantly different when analyzing the images in the sagittal plane only, but the technique allowed demonstration of all aspects using the combined 2D/3D interpretation. Conclusion The NPC may exhibit important anatomical variations, both with regard to morphology and its dimensions. To avoid any potential complications during dentoalveolar surgery, careful preoperative evaluation is required. 3D imaging is recommended to determine canal topography and dimensions, and to assess the individual anterior maxilla's dimensions prior to surgical procedures, such as dental implant insertion or bone augmentation.

Specific Antibodies to Staphylococcus aureus Biofilm Are Present in Serum from Pigs with Osteomyelitis

Abstract: Background The Achilles heel in osteomyelitis is that bacteria, primarily Staphylococcus aureus, grow as a biofilm in the bone lesions. Materials and methods In the present study, we explored the serum level of specific antibodies to S. aurues biofilm in porcine models of osteomyelitis. Results Significantly increased levels of antibodies towards the specific biofilm antigen SA0688 were measured in serum from pigs with S. aureus-associated acute and chronic osteomyelitis 5-7 and 10-14 days after inoculation, respectively. Simultaneously with raised antibody levels, an increase in serum interleukin 6 (IL 6) levels was also seen. Conclusion The observed biofilm-specific antibody response represents a T-helper cell 17 (Th17) response and potentially a T-helper cell 1 (Th1) response. This is in agreement with previous studies in mice and rabbits speculating that S. aureus induces a Th1- and Th17-biased adaptive immune response, instead of a protective Th2 response, in order to evade the immune system, resulting in a chronic infection.

Serum and Urinary Neutrophil Gelatinase-associated Lipocalin Monitoring in Normal Pregnancy Versus Pregnancies Complicated by Pre-eclampsia

Abstract: Aim: Pre-eclampsia is a syndrome characterized by endothelium dysfunction, systemic inflammation, and kidney injury that could be associated with increased levels of neutrophil gelatinase-associated lipocalin (NGAL). We investigated whether serum and urinary NGAL may have a clinical value in defining the severity of pre-eclampsia. Patients and Methods: This cross-sectional case–control study enrolled 18 women with pre-eclampsia matched for gestational age with 22 uncomplicated pregnancies. We evaluated the correlation between NGAL levels and blood pressure and 24-hour proteinuria values by linear regression. Results: Linear regression disclosed a positive and significant correlation between urinary NGAL and 24-hour proteinuria. Serum NGAL appeared to be higher, but not significantly different, in severe pre-eclampsia. Conclusion: These preliminary data indicate that NGAL may correlate with an inflammatory renal involvement in severe pre-eclampsia. Further studies would be useful to better estimate the clinical value of an NGAL increase for evaluating the possibility of delivery induction.

The Radical Scavenging Activity and Cytotoxicity of Resveratrol, Orcinol and 4-Allylphenol and their Inhibitory Effects on Cox-2 Gene Expression and Nf-κb Activation in RAW264.7 Cells Stimulated with Porphyromonas gingivalis-fimbriae

Abstract: BACKGROUND/AIM Resveratrol is a polyphenol with efficient anti-oxidative and anti-inflammatory activity. To clarify the molecular mechanism responsible for its anti-inflammatory action, we investigated the radical scavenging activity, cytotoxicity and anti-inflammatory activity of resveratrol and its related compounds, orcinol and 4-allylphenol. MATERIALS AND METHODS The radical scavenging activities of these compounds were determined by the DPPH (2,2'-diphenyl-1-picrylhydrazyl) assay and their cytotoxicities against RAW264.7 cells were determined using a cell-counting kit (CCK-8). The inhibitory effects of these compounds on cyclooxygenase-2 (Cox2) expression in RAW264.7 cells stimulated with Porphyromonas gingivalis (Pg) fimbriae were also determined using real-time polymerase chain reaction and western blot analysis, while inhibition of the fimbria-stimulated activation of nuclear factor-kappa B (Nf-κb) was evaluated using western blot analysis and enzyme-linked immunosorbent assay-like microwell colorimetric transcription factor activity assay, respectively. The quantum chemical parameters were calculated on the basis of the density function theory (DFT) BLYP/6-31G*. RESULTS DPPH radical scavenging activity declined in the order resveratrol > orcinol > 4-allylphenol. The cytotoxicity of the compounds was in the order 4-allylphenol > resveratrol > orcinol. The inhibitory effect on Pg fimbria-stimulated Cox2 expression and Nf-κb activation was enhanced by resveratrol-alone. Resveratrol showed high electronegativity (χ) and softness (σ) values, as determined by quantum chemical calculations. CONCLUSION Resveratrol exerts potent anti-inflammatory activity in RAW264.7 cells stimulated with Pg-fimbriae and may be applicable as a therapeutic agent for inflammatory periodontal disease as a manifestation of systemic disease.

The Spectrum of Infectious Diseases in Kidney Transplantation: A Review of the Classification, Pathogens and Clinical Manifestations.

Abstract: Kidney transplantation is the treatment-of-choice for a significant number of patients with end-stage renal disease. Renal transplant recipients (RTRs) benefit from a longer life expectancy, with a better quality of life. Despite, recent accomplishments in the field of kidney transplantation, both short- and long-term, surgical and medical complications still exist. Among these complications, cardiovascular disease, carcinogenesis and infections are the most important. Infectious diseases constitute the most common complications after renal transplantation and the second most common cause of death among RTRs with a functioning graft. Theoretically, all infectious pathogens could cause disease in immunocompromised RTRs, yet among these, one could identify more important ones, such as the Enterobacteriaceae, causing urinary tract infections; pneumonia due to Pneumocystis jirovecii; Candida species which cause invasive fungal infections; herpes viruses; hepatitis viruses and parasites. Early diagnosis and effective treatment are key elements in salvaging both the allograft and the patient. However, clinical manifestations and diagnosis of such infectious diseases are not easily identified due to the altered state of immune response of the RTR. Thus, apart from possessing a deep knowledge of the etiology and the treatment options in each case, transplant physicians should also always remain alert when dealing with RTRs.

Immunosuppression and Abdominal Wall Defects: Use of Autologous Dermis.

Abstract: Aim The purpose of the present study was to analyze the use of autologous dermis compared to non-biological meshes in repair of abdominal wall defects in patients with redundant skin and immunosuppression. Patients and methods Eighteen patients with abdominal wall defects, immunosuppression and with redundant skin were divided into two groups: Group A consisted of 11 patients treated with autologous dermis. In these patients, autologous dermis was isolated from the amount of skin resected from the inferior abdominal region that was used as a reinforcement of fascial margins above the defect. Group B consisted of seven patients treated with non-biological meshes We evaluated the infection rate of these groups. Results The infection rate was significantly lower in group A patients. Conclusion The transplantation of autologous dermis as a reinforcement for the reconstruction of abdominal wall defects is reasonable for highly selected patients. The use of the dermis was proven useful and we found a lower rate of infection and recurrence.

Aortic Valve Endocarditis Caused by Abiotrophia defectiva: Case Report and Literature Overview

Abstract: UNLABELLED Abiotrophia defectiva or nutritionally variant Streptococcus (NVS) are a rare but important cause of infectious endocarditis, with high rates of bacteriological failure and mortality. We report the case of a 74-year-old man admitted for fever, fatigue and general malaise in the absence of any underlying cardiac, immunosuppressive illness and previous dental manipulations. Transthoracic and transesophageal echocardiogram revealed bacterial vegetation and significant aortic stenosis and regurgitation. Initial blood culture reported gram-positive cocci in chains, subsequently identified as A. defectiva. The patient completed 6 weeks of antibiotic therapy with ampicillin, with a significant decrease of serum inflammatory markers. He refused cardiac surgery and had relapsing endocarditis with positive blood culture for the same pathogen. The patient was then submitted to double-valve cardiac surgery, obtaining a prompt resolution of clinical signs and symptoms, without other relapse or any complications. CONCLUSION Infectious diseases caused by A. defectiva are extremely rare illnesses. Due to the difficult isolation of the pathogen and the slow clinical progression, clinicians should be aware of this bacterium when dealing with blood culture-negative infective endocarditis.

Seasonal variation of serum vitamin D among Greek female patients with osteoporosis.

Abstract: Background/aim Vitamin D (vitD) levels are positively associated with bone health and seasonality affects serum vit D. The aim of the study was to investigate the degree of seasonal variation on 25-hydroxyvitamin D (25(OH)D) serum levels in a population-based cohort of post-menopausal women with osteoporosis. Patients and methods Serum levels of 25(OH)D were assessed in 596 patients (mean age=65.3 years; standard deviation (SD)=9.4) in different time points over a period of 2.5 years. Results The minimum 25(OH)D serum levels were observed in March (13.4±9.5 ng/ml) and the maximum levels in August, September and October (29.1±16.1, 28.9±12 and 28.4±8.9 ng/ml, respectively). The prevalence of vitD deficiency, insufficiency and sufficiency in March was 76.5, 15.7 and 7.8%, respectively. On the contrary, the highest prevalence of vitD sufficiency was observed in August, September and October (38.1%, 45.3% and 46.5%, respectively). Conclusion Seasonal variations should be considered when measuring for 25(OH)D serum levels and treating vitD deficiency.

Influence of Palmatine on Bone Metabolism in Ovariectomized Mice and Cytokine Secretion of Osteoblasts.

Abstract: Background/Aim: Receptor activator of nuclear factor-ĸB ligand (RANKL) and osteoprotegerin (OPG), regulate the cognate receptor RANK on osteoclast precursor cells. Herein we examined the inhibitory effects of palmatine on bone metabolism using ovariectomized (OVX) mice. Materials and Methods: The first experimentaI set was designed to histologically and biochemically examine mice randomly divided into four groups: sham-operated, OVX, and OVX-palmatine intake groups (1 mg/kg and 10 mg/kg). The second experimental set examined the influence of palmatine on osteoblast-like cells in vitro. Results: Palmatine caused significant suppression of osteoclast numbers in tissues. In palmatine-treated mice, RANKL and OPG expression decreased. In the culture supernatant of MC3T3- E1 cells, RANKL and OPG levels were significantly reduced by palmatine addition. Conclusion: Palmatine may attenuate osteoclast differentiation through inhibition of RANKL and OPG expression by osteoblasts. Therefore, palmatine might be a candidate anti-resorptive agent for osteoporosis therapy. Osteoporosis is a major global health problem, resulting in more than 200 million osteoporotic fractures each year worldwide, including 1.6 million hip fractures (1). Osteoporosis affects both men and women of all races. White and Asian women, particularly older post-menopausal women, are at the highest risk. Bone mass peaks in the twenties and gradually decreases after that due to bone metabolism. In women, bone resorption accelerates with menopause due to a decline in the estrogen level. During menopause, a high turnover rate of bone loss characterized by increased osteoclast activity is observed. Later, the system swings back to normalized osteoclast activity with a slight deficit in osteoblast function and a low turnover of bone loss (1). Bisphosphonates are commonly used for the treatment of osteoporosis and to improve the quality of life in a vast majority of patients (2). However, a number of reports have described necrotic maxillofacial bone lesions in patients who have received high dosage of bisphosphonates, particularly when the patient undergoes subsequent dental procedures. Despite numerous investigations, the problem of the treatment and prevention of osteoporosis remains unresolved. Palmatine is a quaternary protoberberine alkaloid. It is typically yellow in color and is an active constituent of a number of plants, such as Coptidis rhizoma (3). The alkaloid has been used in the treatment of jaundice, dysentery, hypertension, inflammation, and liver-related diseases (4). It has been reported that berberine, an isoquinoline alkaloid which is a close structural analog of palmatine, inhibited bone loss in an osteoporosis model (5). However, there are few reports on the effect of palmatine on bone loss. Ovariectomy has been extensively used to establish an experimental animal model for post-menopausal osteoporosis. Ovariectomized animals exhibited a significant decrease in bone mineral density (BMD) and bone formation (6). Therefore, in order to evaluate the osteoimmunological effects of a component derived from the crude drug on post- menopausal osteoporosis, we examined whether palmatine could prevent bone loss in OVX mice in vivo and control cytokine secretion of osteoblasts in vitro.

Comparison between single PCR and nested PCR in detection of human papilloma viruses in paraffin-embedded OSCC and fresh oral mucosa.

Abstract: Background: Infection with human papilloma virus (HPV) has been implicated as one of the risk factors for the development of oropharyngeal cancer. Many different HPV tests exist, and information re ...

Radiation Therapy for Metastatic Spinal Cord Compression in Patients with Hepatocellular Carcinoma.

Abstract: Aim This is the first study to investigate patients with metastatic spinal cord compression (MSCC) from hepatocellular carcinoma (HCC). Patients and methods Eight patients were analyzed with regard to survival and motor dysfunction. Results Out of seven factors (age, affected vertebrae, ambulatory status, bone lesions, other distant metastases, time developing motor dysfunction, performance score) ambulatory status (p=0.005) and distant metastases (p=0.032) had a significant influence on survival. Both factors were used as a predictive tool (points: not ambulatory 0, ambulatory 1, distant metastases 0, no distant metastases 1). Total scores were 0, 1 or 2 points. Three-month survival rates were 0%, 67% and 100%, six-month survival rates 0%, 0% and 100%. Progression of motor dysfunction was prevented in 63% of patients; time developing motor deficits showed a trend (p=0.08). Conclusion Many patients with MSCC from HCC have a short survival, which can be predicted with a new tool. Radiation therapy can stop progression of motor dysfunction.

Genetic and Apoptotic Changes in Lungs of Mice Flown on the STS-135 Mission in Space

Abstract: AIM The goal of the study was to evaluate changes in lung status due to spaceflight stressors that include radiation above levels found on Earth. MATERIALS AND METHODS Within hours after return from a 13-day mission in space onboard the Space Shuttle Atlantis, C57BL/6 mice (FLT group) were euthanized; mice housed on the ground in similar animal enclosure modules served as controls (AEM group). Lung tissue was collected to evaluate the expression of genes related to extracellular matrix (ECM)/adhesion and stem cell signaling. Pathway analysis was also performed. In addition, immunohistochemistry for stem cell antigen-1 (SCA-1), the terminal deoxynucleotidyl transferase dUTP nick-end labeling (TUNEL) assay for apoptosis, and staining for histological characteristics were performed. RESULTS There were 18/168 genes significantly modulated in lungs from the FLT group (p<0.05 vs. AEM); 17 of these were up-regulated and one was down-regulated. The greatest effect, namely a 5.14-fold increase, was observed on Spock1 (also known as Spark/osteonectin), encoding a multi-functional protein that has anti-adhesive effects, inhibits cell proliferation and regulates activity of certain growth factors. Additional genes with increased expression were cadherin 3 (Cdh3), collagen, type V, alpha 1 (Col5a1), integrin alpha 5 (Itga5), laminin, gamma 1 (Lamc1), matrix metallopeptidase 14 (Mmp14), neural cell adhesion molecule 1 (Ncam1), transforming growth factor, beta induced (Tgfbi), thrombospondin 1 (Thbs1), Thbs2, versican (Vcan), fibroblast growth factor receptor 1 (Fgfr1), frizzled homolog 6 (Fzd6), nicastrin (Ncstn), nuclear factor of activated T-cells, cytoplasmic, calcineurin-dependent 4 (Nfatc4), notch gene homolog 4 (Notch4) and vang-like 2 (Vangl2). The down-regulated gene was Mmp13. Staining for SCA-1 protein showed strong signal intensity in bronchiolar epithelial cells of FLT mice (p<0.05 vs. AEM). TUNEL positivity was also significantly higher in the FLT mice (p<0.05 vs. AEM), but no consistent histological differences were noted. CONCLUSION The results demonstrate that spaceflight-related stress had a significant impact on lung integrity, indicative of tissue injury and remodeling.

Anti-inflammatory activity of the artificial antioxidants 2-tert-butyl-4-methoxyphenol (BHA), 2,6-di-tert-butyl-4-methylphenol (BHT) and 2,4,6-tri-tert-butylphenol (TBP), and their various combinations.

Abstract: Background/aim The artificial complex phenols, 2-tert-butyl-4-methoxyphenol (BHA), 2,6-di-tert-butyl-4-methylphenol (BHT) and 2,4,6-tri-tert-butylphenol (TBP) exert efficient antioxidant activity; however, they are considerable toxic and potentially tumor-promoting. These phenols, particularly in combinations, have enhanced antioxidant activity due to synergistic interactions and produce bioactive intermediates such as quinone methide. We investigated the anti-inflammatory activity of BHA, BHT and TBP, and combinations of BHT/BHA (in molar ratios of 1:1, 1:2, 1:3 and 2:1), BHT/TBP (1:1), and BHA/TBP (1:1), using gene-expression systems for cyclooxygenase-2 (Cox2) and tumor necrosis facto-alpha (Tnfa) in RAW264.7 cells. Materials and methods The inhibitory effects of BHA, BHT and TBP on expression of Cox2 and Tnfa genes upon stimulation with Escherichia coli lipopolysaccharide (LPS) or Porphyomonas gingivalis (Pg) fimbriae were determined using real-time polymerase chain reaction. Results The inhibitory effect on expression of Cox2 and Tnfa genes upon stimulation with LPS and fimbriae was greatly enhanced by the combination of two antioxidants (molar ratio 1:1), BHT/BHA. In addition, that of the Cox2 gene, but not of Tnfa gene was slightly enhanced by a combination of equimolar BHT/TBP and BHA/TBP. None of the antioxidants alone exerted any anti-inflammatory activity upon stimulation with LPS, but a slight anti-inflammatory activity was observed upon stimulation with Pg fimbriae. The inhibitory effect of the BHT/BHA combination on expression of Cox2 mRNA upon stimulation with LPS was investigated at afferent molar ratios, and a molar ratio of 1:1 was found to have considerably less effect than a molar ratio of 1:2 or 2:1. The 1:3 combination had no effect. Conclusion The combination of BHT and BHA at a molar ratio of 0.5-2 exerts potent anti-inflammatory activity. This anti-inflammatory activity on the generation of inflammatory mediators in LPS-activated RAW264.7 cells may be attributable to complex synergistic antioxidant activity of the combination of BHT and BHA. Our results suggest the potential usefulness of the BHT/BHA combination at an appropriate molar ratio as an antioxidant in foods and pharmaceuticals, whereas either antioxidant alone is unlikely to be effective.