The impact of Sambucus nigra L. extract on inflammation, oxidative stress and tissue remodeling in a rat model of lipopolysaccharide-induced subacute rhinosinusitis.

Rhinosinusitis is a common disorder related to inflammation of paranasal sinuses and nasal cavity mucosa. Herbal medicines could be an option in the treatment of rhinosinusitis due to their anti-inflammatory and anti-oxidative properties. The study aims to investigate the effect of intranasal Sambucus nigra L. subsp. nigra (SN) extract against inflammation, oxidative stress, and tissue remodeling in nasal and sinus mucosa, but also in serum, lungs, and brain, in Wistar rat model of subacute sinonasal inflammation induced by local administration of lipopolysaccharides (LPS), from Escherichia Coli. The cytokines (TNF-α, IL-1ß, IL-6) and oxidative stress (malondialdehyde) in nasal mucosa, blood, lungs, and brain were analyzed. In addition, a histopathological examination was performed, and NF-kB, MMP2, MMP9, TIMP1 expressions were also evaluated in nasal mucosa. Both doses of LPS increased the production of cytokines in all the investigated tissues, especially in the nasal mucosa and blood (p < 0.01 and p < 0.05), and stimulated their secretion in the lungs, and partially in the brain. Malondialdehyde increased in all the investigated tissues (p < 0.01 and p < 0.05). In parallel, upregulation of NF-kB and MMP2 expressions with downregulation of TIMP1, particularly at high dose of LPS, was observed. SN extract reduced the local inflammatory response, maintained low levels of IL-6, TNF-α, and IL-1ß. In lungs, SN reduced all cytokines levels while in the brain, the protective effect was noticed only on IL-6. Additionally, SN diminished lipid peroxidation and downregulated NF-kB in animals exposed to a low dose of LPS, with increased TIMP1 expression, while in animals treated with a high dose of LPS, SN increased NF-kB, MMP2, and MMP9 levels. In conclusion, SN extract diminished the inflammatory response, reduced generation of reactive oxygen species (ROS) and, influenced MMPs expressions, suggesting the benficial effect of SN extract on tissue remodeling in subacute rhinosinusitis and on systemic inflammatory response.

Factors Contributing to Non-Exclusive Breastfeeding in Primigravid Mothers

Aims We aimed to examine the factors contributing to non-exclusive breastfeeding in primigravid mothers in a large Irish tertiary maternity hospital. Methods This was a retrospective cohort study carried out at the Rotunda Hospital, Dublin, Ireland. Maternal demographics, antenatal, perinatal, delivery-related information and neonatal outcomes were collected and analysed. Results 569 eligible mothers were delivered during the study period. Out of the 416 mothers intending to breastfeed, 278 (67%) mothers were exclusively breastfeeding at discharge. On univariate analysis, a higher body mass index, unemployment, an Asian background, gestational diabetes, antenatal steroids, low birth weight and hypernatremia were all associated with non-exclusive breastfeeding (all p<0.05). On logistic regression, only gestational diabetes, a birthweight < 2500 grams and hypernatremia remained significantly associated with non-exclusive breastfeeding on discharge. Conclusion Addressing barriers to breast feeding through antenatal and early neonatal education, counselling and support, by qualified healthcare personnel may increase the number of infants exclusively breastfeeding on discharge.

Gold nanoparticles phytoreduced with Cornus mas extract mitigate some of gliadin effects on Caco-2 cells.

Celiac disease (CD) is an autoimmune condition that occurs in genetically predisposed people where the ingestion of gluten produces damage in the small intestine. The treatment accepted until now is a strict gluten free diet. This implies the need for novel or adjuvant treatments, in addition to the standard of care. The present study aimed to assess the effect of gold nanoparticles phytosynthesized with Cornus mas extract (AuCM) compared to Cornus mas extract (CM) and luteolin (LT) on Caco-2 cells, exposed or not to gliadin. Ultraviolet-visible spectroscopy and transmission electron microscopy were used for the characterization of AuCM. Measured cellular outcomes included oxidative stress markers (malondialdehyde level, catalase and superoxide dismutase activities), inflammatory response and cellular signaling and transcription factors involved in apoptosis (NFκB, pNFκB, NOS2, TNF-α, TRAIL, Bax, Bcl-2, p53). The internalization of gold nanoparticles in cells was evidenced by transmission electron microscopy (TEM). The gliadin administration induced oxidative stress, improved the activity of antioxidants enzymes, increased NOS2 and NFκB expressions and reduced pNFκB/NFκB ratio. In addition, gliadin enhanced TRAIL and Bcl-2 levels and reduced p53 expression in Caco-2 cells. The pretreatment with AuCM, CM extract and LT diminished oxidative stress and reduced NOS2 activity. AuCM and CM treatment amplified the expression of p53 and pNFκB/NFκB ratio and diminished Bcl-2, NFκB and pNFκB, especially AuCM. The results obtained confirmed that AuCM mitigate some of gliadin effects on Caco-2 cells through modulation of oxidative stress and inflammation.

The effect of Tropaeolum majus L. on bacterial infections and in vitro efficacy on apoptosis and DNA lesions in hyperosmotic stress.

Tropaeolum majus L. (T. majus) or nasturtium is a medicinal plant widespread in the areas with temperate climate, commonly used in culinary and in traditional medicine due to therapeutic properties. In the last few years, various effects of the flowers and leaves of this plant have been studied, but their benefits are not fully known. The aim of the study was to identify the phenolic compounds from T. majus edible flowers in relation with its antioxidant capacity and the antimicrobial activity against different bacteria and Candida albicans. In addition, the impact of natural extract on oxidative stress, inflammation and apoptosis was analysed on human umbilical vein endothelial cells (HUVECs) exposed to normotonic and hypertonic conditions. The major phenolic acids, identified by HPLC-RP with UV detection, were gallic acid, caffeic acid and p-coumaric and predominant flavonoids were quercetin, epicatechin and luteolin. The both fractions of T. majus were rich sources of polyphenols with marked antioxidant activity, evidenced by TEAC or DPPH methods. The extract exhibited a week antibacterial effect on some strains of streptococcus, without antifungal or antibacterial effect on gram negative bacteria. T. majus extract increased the p53 and Bcl-2 expressions and diminished the DNA lesions indicating the protective and antiapoptotic effects in vitro, on endothelial cells exposed to hyperosmotic stress. These experimental findings suggest that T. majus can exert some protection against bacterial infections and reduce apoptosis and DNA lesions in hypertonic conditions.

Characterization and biological effects of Hypericum extracts on experimentally-induced - anxiety, oxidative stress and inflammation in rats.

Anxiety disorders can associate with oxidative stress and immune system alterations. Our study aimed to chemically analyze Hypericum maculatum (HM) and Hypericum perforatum (HP) dry extracts and to evaluate their effects along with quercetin (Q), on brain oxidative stress biomarkers: malondialdehyde (MDA), catalase (CAT) and superoxide dismutase (SOD), inflammatory cytokines: interleukin-1α, (IL-1α), IL-1ß, regulated on activation normal T cell expressed and secreted (RANTES), interferon (IFN), monocyte chemoattractant protein-1 (MCP1), macrophage inflammatory protein (MIP) and serum corticosterone levels. Nuclear transcription factor κB (NFκB) signaling pathway in the hippocampus and frontal lobe in rats with N-methyl-9H-pyrido[5,4-b]indole-3-carboxamide (FG-7142) experimental-induced anxiety were also investigated. The chemical analyses of total hypericins were performed by spectrophotometric analysis and hypericin, hyperforin and polyphenols derivatives were quantified by chromatographic methods. The animals were divided in 6 groups: carboxymethylcellulose 2% (CMC); CMC + FG; alprazolam (APZ) + FG; Q + FG; HM + FG; HP + FG. APZ (0.08 mg/kg b.w.), Q (30 mg/kg b.w.), HM and HP (350 mg/kg b.w.) were orally administered for 21 days. FG (7.5 mg/kg b.w.) was intraperitoneally (i.p.) injected in a single dose. Q and hypericum extracts (HpE) exerted anti-inflammatory (decreased IL-1α, IL-1ß, MCP1, IFN and MIP mainly in hippocampus) and antioxidant effects (decreased MDA levels, increased CAT and SOD activity), enhanced NFκB and pNFκB expressions in the brain and reduced serum corticosterone levels. Our findings suggest that HpE may improve anxiety-like behavior, offer brain protection by modulation of oxidative stress and inflammation, and can contribute to overall biological activity of natural compounds-rich diet.

Free radicals, ceruloplasmin, and copper concentration in serum and aortic tissue in experimental atherosclerosis.

It has been suggested that the increase in concentration of blood lipid peroxides may be a risk factor for atherosclerosis. Our research was conducted to study the effect of cholesterol feeding on lipid peroxides, ceruloplasmin, HDL-cholesterol, serum unesterified fatty acids, and copper concentration in serum and aortic tissue in rats. In the animals fed a cholesterol-supplemented diet, the mean values of serum lipid peroxide, ceruloplasmin, serum copper, and unesterified fatty acids were increased significantly (p < 0.01) as compared to the control group. At the same time, HDL-cholesterol concentration was significantly decreased in rats fed a diet enriched with cholesterol (p < 0.01) compared to the control group. A significant decrease of copper concentration in aorta was also observed in these animals versus controls. Correlations between ceruloplasmin and lipid peroxides as well as copper were statistically significant in these animals. At the same time, antioxidant activity in blood was two times higher as compared to controls. Results of this study provide the support for applying these determinations in atherosclerosis monitoring.

[Ascites--a complication of chronic alcoholic pancreatitis]. / Ascita--complicatie a pancreatitei cronice etanolice.

We present the case a 44 year old patient previously diagnosed with chronic alcoholic pancreatitis with pancreatic ascites during hospitalization in the Gastro-Enterology department. As the conservative therapy performed for 21 days was not effective in diminishing the ascites, the patient was admitted in our Surgical Department and scheduled for surgical intervention. He was operated and we discovered a small dimension cyst (7/4 cm) developed in the body and tail of the pancreas, fistulized in the peritoneal cavity through an outlet positioned below the insertion of the mesocolonum transversum, fairly close to the duodeno-jejunal angle. We executed a cysto-jejunal anastomosis by using the first loop of the jejunum, secured with a politer drainage positioned as in WITZEL technique and drive out in the left upper quadrant. The postoperative evolution of the patient was difficult, but constantly positive. The patient left the hospital 32 days after the intervention. The clinical and ultrasound follow-up after three months were normal.

In vitro studies on the mechanisms involved in chemoprevention using Calluna vulgaris on vascular endothelial cells exposed to UVB.

The study aims to investigate the mechanisms involved in the in vitro effect of UVB on endothelial vascular cells (HUVECs) pretreated with a photochemopreventive agent, the Calluna vulgaris (Cv) extract. Two concentrations of Cv, below the limit of cytotoxicity IC50 (2.5 and 7.5 µg GAE/ml) and two doses of UVB (50 and 100 mJ/cm(2)) were used. Oxidative stress parameters were quantified at 1 h and 24 h after irradiation and apoptosis, DNA damage and the induction/activation of NF-κB were evaluated at 24 h. UVB exposure led to the formation of lipid peroxides in a dose dependent manner (p<0.001), induced apoptosis, increased the γ-H2AX levels and the activation of NF-κB. Pretreatment with 2.5 µg GAE/ml Cv improved the antioxidant defense, protected against DNA lesions and was able to decrease cellular death at low dose of irradiation. 7.5 µg GAE/ml Cv was prooxidant, favored the formation of DNA lesions, amplified the NF-κB activation UVB-induced (p<0.01) and led to high levels of cellular death. Both doses of Cv inhibited caspase-3 activation. The modulatory effect of Cv extract on endothelial cells exposed to UVB depend on the concentration of Cv used. This study provides insides into the mechanisms triggered by UVB and antioxidants on skin endothelial cells.

Spironolactone and dimethylsulfoxide effect on glucose metabolism and oxidative stress markers in polycystic ovarian syndrome rat model.

Because polycystic ovarian syndrome (PCOS) is a risk factor for type 2 diabetes, the affected women can present frequently prediabetic states such as impaired fasting glycaemia and/or impaired glucose tolerance. The purpose of our study is to explore the effect of antiandrogenic spironolactone on glucose metabolism and oxidative stress (OS) parameters in oestradiol valerate (OV) induced PCOS rat model.72 female Wistar rats were distributed either to PCOS group (n=65, OV dissolved in sesame oil, 5 mg/0.4 ml), or to non-PCOS control group (n=7, sesame oil, 0.4 ml). After a month, ultrasound was performed to assess the ovarian morphology, and the results of an initial oral glucose tolerance test (OGTT) were used to identify the animals with altered glucose metabolism (AGM). Glucose transporter 4 (GLUT4) was evaluated from muscle biopsies, OS parameters were assessed from blood and muscle samples, and ovaries of 3 rats were removed for histopathological examination. Afterwards, the AGM group was divided in a treated PCOS group denoted as Sp+D (per os spironolactone dissolved in DMSO, 2 mg/0.2 ml), and a PCOS control treated with DMSO (0.2 ml). After one month of daily treatment, a final OGTT was performed. GLUT4 and OS parameters were again evaluated and ovaries were removed for histopathological examination.As compared to the values prior to the treatment, Sp+D reversed fasting hyperglycaemia (p<0.001), increased GLUT4 immunoreactivity in the perinuclear compartment (p<0.05) and translocation to plasmalemma (p<0.001) and improved superoxide dismutase (0.001

Hepatic and systemic effects of rosuvastatin on an experimental model of bile duct ligation in rats.

In the early stages of cholestasis, a plethora of mechanisms are considered to contribute to the liver injury: oxidative stress, inflammation, cholangiocytes proliferation and fibrosis. Our study aims to investigate the effects of different doses of rosuvastatin (Ro) on experimental bile duct ligation-induced cholestasis. 40 female Wistar rats were randomly divided into 4 groups (n=10): Sham group (laparotomy); BDL group (subjected to bile duct ligation); BDL group treated with Ro (5 mg/bw daily); BDL group treated with Ro (10 mg/bw daily). After 6 days of treatment, in the day 7 after BDL, the animals were sacrificed and we explored hepato-cytolysis, the seric parameters for cholestasis and oxidative stress in plasma, liver, brain and kidneys. Proliferation was investigated by expression of proliferating cell nuclear antigen (PCNA), while inflammation by liver histology, TNFR2 expression and NF-κB induction and activation. To assess fibrosis, we performed Tricrom-Masson staining, transforming growth factor beta-1 (TGF-ß1) expression, and for myofibroblast activation, we analyzed α-SMA expression. The administration of Ro in early stages of cholestasis proved to have a beneficial effect by decreasing α-SMA. Ro didn't exert systemic oxidative stress effects, but increased hepatocytolysis, oxidative stress and inflammation in the liver and sustained increased levels of pro-fibrotic cytokine TGF-ß1 as well as the number of proliferating cells in ducts and parenchyma. Ro inhibited the induction and the activation of NF-κB, which could be considered a beneficial effect. Further studies must be carried out in order to clearly investigate the balance between risks and benefits for Ro administration in early stages of cholestasis.

Possible in vivo mechanisms involved in photodynamic therapy using tetrapyrrolic macrocycles.

Photodynamic therapy (PDT) mediated by oxidative stress causes direct tumor cell damage as well as microvascular injury. To improve this treatment new photosensitizers are being synthesized and tested. We evaluated the effects of PDT with 5,10,15,20-tetrakis(4-methoxyphenyl)-porphyrin (TMPP) and its zinc complex (ZnTMPP) on tumor levels of malondialdehyde (MDA), reduced glutathione (GSH) and cytokines, and on the activity of caspase-3 and metalloproteases (MMP-2 and -9) and attempted to correlate them with the histological alterations of tumors in 3-month-old male Wistar rats, 180 ± 20 g, bearing Walker 256 carcinosarcoma. Rats were randomly divided into five groups: group 1, ZnTMPP+irradiation (IR) 10 mg/kg body weight; group 2, TMPP+IR 10 mg/kg body weight; group 3, 5-aminolevulinic acid (5-ALA+IR) 250 mg/kg body weight; group 4, control, no treatment; group 5, only IR. The tumors were irradiated for 15 min with red light (100 J/cm², 10 kHz, 685 nm) 24 h after drug administration. Tumor tissue levels of MDA (1.1 ± 0.7 in ZnTMPP vs 0.1 ± 0.04 nmol/mg protein in control) and TNF-α (43.5 ± 31.2 in ZnTMPP vs 17.3 ± 1.2 pg/mg protein in control) were significantly higher in treated tumors than in controls. Higher caspase-3 activity (1.9 ± 0.9 in TMPP vs 1.1 ± 0.6 OD/mg protein in control) as well as the activation of MMP-2 (P < 0.05) were also observed in tumors. These parameters were correlated (Spearman correlation, P < 0.05) with the histological alterations. These results suggest that PDT activates the innate immune system and that the effects of PDT with TMPP and ZnTMPP are mediated by reactive oxygen species, which induce cell membrane damage and apoptosis.

The dynamics of reactive oxygen species in photodynamic therapy with tetra sulfophenyl-porphyrin.

Photodynamic therapy (PDT) is a promising therapy especially in skin cancer, using the systemic administration of a photosensitizer (PS), followed by the local irradiation of the tumor with visible light. The antitumor effects of PDT are determined especially by the generation of cytotoxic reactive oxygen species (ROS). The 5,10,15,20-tetra-sulfo-phenyl-porphyrin (TSPP) is a synthetic photosensitizer, which proved its efficiency in in vitro studies. Our study evaluates the effects of PDT with TSPP upon the tumor levels of ROS and upon the metalloproteinases 2 (MMP2) activities on Wistar male rats bearing 256 Walker carcinosarcoma in correlation with the accumulation of PS in the tumor and with the intratumor histological alterations. The evaluations were performed dynamically, at 3 hours, 6 hours, 24 hours and 14 days after the PDT with TSPP. Our results emphasize that 24 hours after the PDT with TSPP, the ROS generation increases, as revealed by protein carbonyls and malondialdehyde levels and the antioxidant capacity (hydrogen donors, thiol groups) decreases in the tumor tissue. These parameters were correlated with the appearance of the histological disorders. The MMP-2 activity increases exponentially in the 24 hours-14 days post PDT interval. PDT with TSPP offers, in vivo , consistent results regarding ROS generation, MMP2 activation and cytotoxic capacity.

Photochemoprevention of cutaneous neoplasia through natural products.

Non-melanoma skin cancers such as squamous cell carcinoma and basal cell carcinoma are the most common types of human tumors, representing 30% of the new cases of malignancies diagnosed each year. Ultraviolet radiation (UV) from the sun is a major cause of non-melanoma skin cancer in humans. The prevention and mainly the photochemoprevention with natural products represent a simple but very effective strategy in the management of cutaneous neoplasia. Here we review the progress in the research of new and existing agents developed to protect the skin exposed to UV. We also discuss the current state of knowledge on their photosuppression mechanism in humans as well as in animal models, and efficiency in cancer prevention.

Possible in vivo mechanisms involved in photodynamic therapy using tetrapyrrolic macrocycles

Photodynamic therapy (PDT) mediated by oxidative stress causes direct tumor cell damage as well as microvascular injury. To improve this treatment new photosensitizers are being synthesized and tested. We evaluated the effects of PDT with 5,10,15,20-tetrakis(4-methoxyphenyl)-porphyrin (TMPP) and its zinc complex (ZnTMPP) on tumor levels of malondialdehyde (MDA), reduced glutathione (GSH) and cytokines, and on the activity of caspase-3 and metalloproteases (MMP-2 and -9) and attempted to correlate them with the histological alterations of tumors in 3-month-old male Wistar rats, 180 ± 20 g, bearing Walker 256 carcinosarcoma. Rats were randomly divided into five groups: group 1, ZnTMPP+irradiation (IR) 10 mg/kg body weight; group 2, TMPP+IR 10 mg/kg body weight; group 3, 5-aminolevulinic acid (5-ALA+IR) 250 mg/kg body weight; group 4, control, no treatment; group 5, only IR. The tumors were irradiated for 15 min with red light (100 J/cm², 10 kHz, 685 nm) 24 h after drug administration. Tumor tissue levels of MDA (1.1 ± 0.7 in ZnTMPP vs 0.1 ± 0.04 nmol/mg protein in control) and TNF-α (43.5 ± 31.2 in ZnTMPP vs 17.3 ± 1.2 pg/mg protein in control) were significantly higher in treated tumors than in controls. Higher caspase-3 activity (1.9 ± 0.9 in TMPP vs 1.1 ± 0.6 OD/mg protein in control) as well as the activation of MMP-2 (P < 0.05) were also observed in tumors. These parameters were correlated (Spearman correlation, P < 0.05) with the histological alterations. These results suggest that PDT activates the innate immune system and that the effects of PDT with TMPP and ZnTMPP are mediated by reactive oxygen species, which induce cell membrane damage and apoptosis.

Extrahepatic manifestations in hepatitis C virus infection.

The authors present a revision of the literature and also their own experience concerning extrahepatic manifestations in hepatitis C virus infection. A special attention receives the lymphotropism of HCV with mixed cryoglobulinemia and leukocytoclastic vasculitis, lichen planus and porphyria cutanea tarda.