Bioactivity of Juniperus communis essential oil and post-distillation waste: Assessment of selective toxicity against food contaminants

Previously chemically characterized Juniperus communis essential oil (EO) and post-distillation waste (PDW) were tested for cytotoxicity and antimicrobial activity against food contaminants. Microdilution assay showed that PDW induced moderate antifungal (minimum inhibitory concentration (MIC) and minimum fungicidal concentration (MFC) values, ranging between 0.118-0.900 mg mL-1), and an antibacterial effect against Listeria monocytogenes (MIC and minimum bactericidal concentration (MBC) were 0.39 and 0.74 mg mL-1, respectively). Combinations of EO/PDW with selected antibiotics induced synergistic antilisterial activity in the checkerboard assay. The MTT assay determined that cytotoxicity against colon cancer cells was high for the EO but negligible for PDW (IC50 values were 0.087-0.106 and 1.450-6.840 mg mL-1, respectively). The selectivity indices indicated high selectivity of PDW against tested fungi and L. monocytogenes. In the adhesion-inhibition assay, PDW reduced in vitro adhesion of L. monocytogenes to colon cells (29-62% of inhibition). In conclusion, PDW exhibited an antimicrobial effect against important food spoilage and poisoning fungi and L. monocytogenes, and also reduced in vitro adhesion of L. monocytogenes to colon cells. The results indicate that J. communis PDW could be considered as natural preservative against food spoilage and poisonous fungi, and as an adjuvant to conventional therapy of listeriosi

The role of heme oxygenase 1 and NF-kB in interscapular brown adipose tissue: The influence of nitric oxide

The male Mill Hill hybrid hooded rats were divided into three main groups one received L-arginine HCl (2.25%), another Nω-nitro-L-arginine methyl ester (L-NAME HCl 0.01%), in drinking water for 45 days and the third group was a control. All groups additionally were divided into two subgroups: one - housed at 22 ± 1 °C and another - at 4 ± 1 °C. HO-1 and NF-kB were identified using immunohistochemistry. Present data disclosed parallelism between HO-1 and NF-kB immunopositivity in IBAT of rats kept at room and low temperature. At room temperature, immunopositivities of HO-1 and NF-kB were increased in L-arginine-treated group when compared to the control, while the strongest immunopositivities and their translocation into nuclei were detected in L-NAME-treated-group. Cold markedly diminished both HO-1 and NF-kB immunopositivity.Cilj ovog rada bio je da se ispita moguća veza između hem oksigenaze 1 (HO-1) i redoks senzitivnog transkripcionog faktora NF-kB (engl. nuclear factor-kappa B) i to naročito u odnosu na azot oksid (NO) u interskapularnom mrkom masnom tkivu (IMMT) pacova. Mužjaci hibridnog Mill-Hill soja pacova podeljeni su u tri grupe, od kojih je jedna tretirana L-argininom (L-arginin-HCl 2.25%), druga s L-NAME (L-NAME-HCl 0.01%), rastvorenim u pijaćoj vodi, a treća, kontrolna grupa je bila tretirana pijaćom vodom. Sve grupe su dodatno podeljene u dve podgrupe koje su tokom eksperimenta boravile na sobnoj (22 ± 1°C), odnosno niskoj temperaturi (4 ± 1°C). Eksperiment je trajao 45 dana. HO-1 i NF-kB su detektovani imunohistohemijski. U ovom radu detektovane su paralelne promene HO-1 i NF-kB imunopozitivnosti u IMMT-u životinja aklimiranih na sobnu i nisku temperaturu. Na sobnoj temperaturi, L-arginin tretman indukovao je povećanje HO-1 i NF-kB imunopozitivnosti u odnosu na kontrolu dok je najjača imunopozitivnosti kao i njena translokacija u nukleuse detektovana u L-NAME tretiranoj grupi. Niska temperatura drastično je smanjila i HO-1 i NF-kB imunopozitivnost. Rezultati ovog rada pokazali su paralelne promene HO-1 i NF-kB imunopozitivnosti u odgovoru na redoks zavisne mehanizme koji uključuju NO i omogućili uspostavljanje bliske i isprepletane veze između NO, HO-1 i NF-kB signalnih puteva u redoks regulaciji IMMT-a.Projekat ministarstva br. 14305

Specific expressional pattern of HO-1 and HO-2 in rat pancreas: Role in experimentally induced diabetes mellitus

The goal of this study was to elucidate expression of heme oxygenase inducible (HO-1) and constitutive (HO-2) isoforms in normal and diabetic pancreas and possible role of their catalytic product - carbon monoxide (CO) in this tissue. The male Mill Hill hybrid hooded rats were divided into two groups: diabetic and non-diabetic. The single dose of alloxan (120 mg/kg) was used for induction of insulin-dependent diabetes mellitus. Both groups were additionally separated in three subgroups: one, treated 12 days with L-arginine HCl (2.25%), another with Nω-nitro-L-arginine methyl ester (L-NAME HCl, 0.01%) and the third was control. HO isoforms were detected by immunohistochemistry. Strong HO-2 immunopositivity occurred in endocrine pancreas of non-diabetic control rats while weaker staining with HO-1 antibody was detected in this group. In diabetic control, as well as in non-diabetic and diabetic treated groups, isoforms specific changes of immunopositivity were observed. Regardless of group, strong immunoreactivity of one isoform was associated with weaker immunoreactivity of another. In conclusion, there is an inverse expressional pattern of HO-1 and HO-2 in normal as well as in diabetic pancreas, which enables maintaining of HO catalytic activity in this tissue. These results suggest that HO/CO pathway might be of importance for pancreatic function.Cilj ovog istraživanja bio je da se ispita ekspresija hem oksigenaze inducibilne (HO-1) i konstitutivne (HO-2) izoforme u normalnom i dijabetičnom pankreasu i moguća uloga njihovog katalitičkog produkta-ugljen monoksida (CO) u ovom tkivu. Mužjaci hibridnog Mill Hill soja pacova su podeljeni u dve grupe: bez dijabetesa i sa dijabetesom. Insulin zavisni diabetes mellitus indukovan je jednom dozom aloksana (120 mg/kg). Obe grupe su dodatno bile podeljene u tri podgrupe: jedna je tretirana 12 dana L-argininom HCl (2.25%) druga s Nω-nitro-L-arginin metil estar (L-NAME HCL, 0.01%) a treća je bila kontrola. HO izoforme su detektovane imunohistohemijski. Kod kontrolnih životinja bez dijabetesa je zabeležena jaka HO-2 imunopozitivnost u endokrinom pankreasu dok je imunoreakcija na HO-1 slabija. Imunopozitivnost HO-1 i HO-2 se menja u dijabetesu i pod dejstvom tretmana pri čemu se odgovor ove dve izoforme na njih međusobno razlikuje. Nezavisno od grupe, povećana imunoreakcija jedne izoforme je vezana sa smanjenom imunoreakcijom druge. Iz rezultata istraživanja proizilazi da postoji inverzan obrazac ekspresije HO-1 i HO-2 u normalnom kao i u dijabetičnom pankreasu što omogućava održanje katalitičke aktivnosti HO u ovom tkivu. Ovi rezultati ukazuju na to da HO/CO sistem može biti od posebnog značaja za funkcionisanje pankreasa.Projekat ministarstva br. 14305

NO modulates the molecular basis of rat interscapular brown adipose tissue thermogenesis

Molecular mechanisms underlying interscapular brown adipose tissue (IBAT) thermogenesis were elucidated. Namely, gene and/or protein expression of uncoupling protein 1 (UCP1), peroxisome proliferator-activated receptor gamma (PPAR gamma), PPAR gamma-coactivator-1 alpha (PGC-1 alpha), vascular endothelial growth factor (VEGF) and proliferating cell nuclear antigen (PCNA) - key molecules that regulate thermogenesis-related processes - mitochondriogenesis, angiogenesis and IBAT hyperplasia, in rats subjected to cold (4 +/- 1 degrees C) for 1, 3, 7, 12, 21 and 45 days were investigated. Particularly, to examine influence of nitric oxide (NO) on IBAT thermogenic-program, cold-exposed animals were treated by L-arginine or N(omega)-nitro-L-arginine-methyl ester (L-NAME). Related to control (22 +/- 1 degrees C), cold induced time-coordinated UCP1, PPAR gamma and PGC-1 alpha transcriptional activation accompanied by PCNA activation and increased VEGF immunolabeling that correlate with endothelial NO synthase (eNOS) transcriptional activation suggesting NO involvement in these thermogenic-factors activation. Observed molecular changes were translated into increased mitochondrialremodeling, angiogenesis, and IBAT hyperplasia. L-Arginine augmented and prolonged cold-induced increase of eNOS, inducible NOS and thermogenic-molecules expression, IBAT nerve supply, vascularity, hyperplasia and mitochondrial-remodeling, while L-NAME had an opposite effects. Results show that NO improves thermogenesis-related mitochondriogenesis, angiogenesis and tissue hyperplasia, positively affecting molecular basis of these processes, suggesting that NO is an essential regulator of IBAT thermogenic-program operating, at genes, proteins and tissue structure levels. (C) 2010 Elsevier Inc. All rights reserved.Ministry of Science and Technological Development of the Republic of Serbia [143050]; [COST FA0602 Action

Antioxidative defense organization in retroperitoneal white adipose tissue during acclimation to cold-The involvement of L-arginine/NO pathway

1. Retroperitoneal white adipose tissue (RpWAT) antioxidative defense was investigated in untreated, L-arginine-treated and N(omega)-nitro-L-arginine methyl ester (L-NAME)-treated rats kept at 4 +/- 1 degrees C (1, 3, 7, 12, 21 and 45 days) and compared to control rats at 22 +/- 1 degrees C. 2. Cold-acclimation-induced RpWAT weight decrease was accompanied by a decline in glutathione level and increased activity of manganese superoxide dismutaise (MnSOD), glutathione S-transferase (GST), catalase, glutathione peroxidase and glutathione reductase at different time-points. 3. L-arginine accelerated RpWAT weight decrease, the increase in MnSOD and GST activities and the prolonged increase of catalase, MnSOD and GST activities. L-NAME delayed cold-induced catalase activity increase and tissue weight decrease. Prolonged L-NAME-treatment had a similar effect on RpWAT as L-arginine. 4. Results suggest the involvement Of L-arginine/NO pathway in RpWAT oxidative metabolic augmentation induced by cold-acclimation. (C) 2009 Elsevier Ltd. All rights reserved.Ministry of Science and Technological Development, Republic of Serbia [143050

Free radical equilibrium in interscapular brown adipose tissue: Relationship between metabolic profile and antioxidative defense

Interscapular brown adipose tissue (IBAT) hyperplasia involves a new metabolic and structural profile, resulting from acclimation of animals to a cold environment. Cold-induced changes of several antioxidative defense (AD) components in IBAT and their interrelationship with uncoupling protein 1 (UCP1), sympathetic innervation and apoptosis were studied using cold-acclimated adult rat males (4 +/- 1 degrees C, 45 days). Their age-matches were maintained at 22 +/- 1 degrees C serving as the controls. In cold-adapted rats, activities of CuZn- and Mn-superoxide dismutase (SOD) and apoptosis were reduced, while catalase (CAT), glutathione peroxidase (GSH-Px), glutathione S-transferase (GST) activities and glutathione (GSH) content were increased compared to the control. IBAT mass, protein content, plasma free fatty acid (FFA) concentration, sympathetic innervation and UCP1 level were significantly increased in cold-acclimated group compared to the corresponding control. These results suggest that decreased CuZn and MnSOD activities in IBAT represent an adaptive response due to UCP1-induced mitochondrial uncoupling. Additionally, intensive fatty acid oxidation led to an increased H2O2 production which resulted in increased CAT, GSH-Px and GST activities and GSH level. Generally speaking, cold-induced changes of AD in the IBAT are closely connected with newly established metabolic profile in this tissue, thus making an important part of the entire tissue homeostasis including cell survival. (c) 2005 Elsevier Inc. All rights reserved.nul

L-Arginine supplementation induces glutathione synthesis in interscapular brown adipose tissue through activation of glutamate-cysteine ligase expression: The role of nitric oxide

We examined whether nitric oxide (NO) in vivo could induce interscapular brown adipose tissue (IBAT) glutathione synthesis. Data show that NO induces in vivo IBAT glutathione synthesis through activation of glutamate-cysteine ligase (GCL) mRNA and protein expression. This NO effect appeared to be mediated by nuclear factor-kappa B (NF-kappa B) activation. We have also observed a complex series of in vivo cellular responses during chronic inhibition of NO synthesis, suggesting that regulatory pathways unrelated to GCL alteration underlie glutathione level increase induced by N(omega)-nitro-L-arginine methyl ester (L-NAME). In general, glutathione synthesis in IBAT seemed to be finely tuned by NO to provide glutathione for either mediating the effects of NO, or for preventing potential nitrosative stress. (C) 2009 Elsevier Ireland Ltd. All rights reserved.Ministry of Science and Technological Development of the Republic of Serbia [143050

Expressional pattern of nitric oxide synthases in normal and diabetic pancreas

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Alterations in L-arginine-nitric oxide-producing pathway affect antioxidative defense in the rat skin

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Specific expressional pattern of HO-1 and HO-2 in rat pancreas: Role in experimentally induced diabetes mellitus

The goal of this study was to elucidate expression of heme oxygenase inducible (HO-1) and constitutive (HO-2) isoforms in normal and diabetic pancreas and possible role of their catalytic product - carbon monoxide (CO) in this tissue. The male Mill Hill hybrid hooded rats were divided into two groups: diabetic and non-diabetic. The single dose of alloxan (120 mg/kg) was used for induction of insulin-dependent diabetes mellitus. Both groups were additionally separated in three subgroups: one, treated 12 days with L-arginine HCl (2.25%), another with Nω-nitro-L-arginine methyl ester (L-NAME HCl, 0.01%) and the third was control. HO isoforms were detected by immunohistochemistry. Strong HO-2 immunopositivity occurred in endocrine pancreas of non-diabetic control rats while weaker staining with HO-1 antibody was detected in this group. In diabetic control, as well as in non-diabetic and diabetic treated groups, isoforms specific changes of immunopositivity were observed. Regardless of group, strong immunoreactivity of one isoform was associated with weaker immunoreactivity of another. In conclusion, there is an inverse expressional pattern of HO-1 and HO-2 in normal as well as in diabetic pancreas, which enables maintaining of HO catalytic activity in this tissue. These results suggest that HO/CO pathway might be of importance for pancreatic function.Cilj ovog istraživanja bio je da se ispita ekspresija hem oksigenaze inducibilne (HO-1) i konstitutivne (HO-2) izoforme u normalnom i dijabetičnom pankreasu i moguća uloga njihovog katalitičkog produkta-ugljen monoksida (CO) u ovom tkivu. Mužjaci hibridnog Mill Hill soja pacova su podeljeni u dve grupe: bez dijabetesa i sa dijabetesom. Insulin zavisni diabetes mellitus indukovan je jednom dozom aloksana (120 mg/kg). Obe grupe su dodatno bile podeljene u tri podgrupe: jedna je tretirana 12 dana L-argininom HCl (2.25%) druga s Nω-nitro-L-arginin metil estar (L-NAME HCL, 0.01%) a treća je bila kontrola. HO izoforme su detektovane imunohistohemijski. Kod kontrolnih životinja bez dijabetesa je zabeležena jaka HO-2 imunopozitivnost u endokrinom pankreasu dok je imunoreakcija na HO-1 slabija. Imunopozitivnost HO-1 i HO-2 se menja u dijabetesu i pod dejstvom tretmana pri čemu se odgovor ove dve izoforme na njih međusobno razlikuje. Nezavisno od grupe, povećana imunoreakcija jedne izoforme je vezana sa smanjenom imunoreakcijom druge. Iz rezultata istraživanja proizilazi da postoji inverzan obrazac ekspresije HO-1 i HO-2 u normalnom kao i u dijabetičnom pankreasu što omogućava održanje katalitičke aktivnosti HO u ovom tkivu. Ovi rezultati ukazuju na to da HO/CO sistem može biti od posebnog značaja za funkcionisanje pankreasa.Projekat ministarstva br. 14305