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The lysosomal membrane stability test applied on the digestive cells (LMS) and the neutral red lysosomal retention assay (NRR) performed on hemocytes have been evaluated on mussels Mytilus galloprovincialis collected from Thermaikos and Strymonikos gulfs (Nothern Greece) in June and December 2000. The correlation analysis indicated that there was a strong positive relationship between the above mentioned biomarkers in both sampling seasons, supporting the validity of their application in water quality assessment. The evaluation of micronucleus frequency (MN) has been also performed in hemocytes of the same mussels and, according to the results, it requires additional validation before using it as a stress index. The results obtained in parallel by the stress on stress (SOS) technique do not support the application of this biomarker in biomonitoring, showing, however, negative correlation with LMS and NRR in one sampling season. Finally, morphological observations were performed on cryosections stained for the enzyme N-acetyl-beta-hexosaminidase, revealing differences in the epithelial cell-layer thickness, as well as changes in the digestive lysosomal system of mussels, obtained from different sampling sites in the two sampling seasons.

The neutral red lysosomal retention assay (NRR) of the haemocytes, and the acetylcholinesterase activity (AChE) in the haemolymph, the digestive gland, the gills and the mantle/gonad complex have been evaluated on mussels Mytilus galloprovincialis collected from Thermaikos and Strymonikos gulfs (northern Greece) in June and October 2001. The validity of performing the above core biomarkers is supported, firstly by their ability to respond to different pollution levels and, secondly, by the significant linear correlation among them. The evaluation of the micronuclei frequency (MN) has been performed in gill tissue and haemocytes of the same mussels and, according to the results, it needs more research in order its use as stress indices to be validated. In addition, the first results on cAMP levels in the gills, the mantle/gonad complex and the digestive gland, whose concentrations correlated to both, NRR and AChE introduce this signal transduction molecule as a new, promising biomarker.

Abstract The lysosomal membrane stability test applied on the digestive cells (LMS) and the neutral red lysosomal retention assay (NRR) performed on hemocytes have been evaluated on mussels Mytilus galloprovincialis collected from Thermaikos and Strymonikos gulfs (Nothern Greece) in June and December 2000. The correlation analysis indicated that there was a strong positive relationship between the above mentioned biomarkers in both sampling seasons, supporting the validity of their application in water quality assessment. The evaluation of micronucleus frequency (MN) has been also performed in hemocytes of the same mussels and, according to the results, it requires additional validation before using it as a stress index. The results obtained in parallel by the stress on stress (SOS) technique do not support the application of this biomarker in biomonitoring, showing, however, negative correlation with LMS and NRR in one sampling season. Finally, morphological observations were performed on cryosections stained for the enzyme N-acetyl-β-hexosaminidase, revealing differences in the epithelial cell-layer thickness, as well as changes in the digestive lysosomal system of mussels, obtained from different sampling sites in the two sampling seasons. [ABSTRACT FROM AUTHOR]

Values of five biomarkers related to cell stress or pollution were evaluated in tissues of the land snail Eobania vermiculata under starvation or hibernation conditions. The biomarkers applied were lysosomal membrane stability in digestive gland cells (LMS) or in haemocytes (neutral red retention assay (NRR)), acetylcholinesterase activity (AchE; EC 3.1.1.7), metallothionein content (MTs), and cyclic AMP content (cAMP). Three groups of snails were studied that were kept under starvation, hibernation and normal conditions, respectively. The results indicated statistically lower values of LMS and NRR in snails kept under starvation or hibernation compared to control ones. Higher values of AChE activity were measured in snails under hibernation compared to controls. MT contents were statistically higher in snails under starvation compared to controls. Measurement of cAMP contents showed no significant differences among the tested groups. The values of the first four biomarkers may be affected by factors other than pollution, such as starvation or hibernation. Therefore, these factors should be taken into consideration when biomonitoring studies are performed in time intervals of hibernation or starvation., (Copyright © 2013 Elsevier Inc. All rights reserved.)

This study investigated the impact of olive mill wastewater (OMW) as a pollutant of the marine environment, via the detection of stress indice alterations in mussels Mytilus galloprovincialis. Due to the absence of data concerning the levels of OMW in the receiving waters, mortality test (96h) was first performed in order to estimate the range of OMW concentration where no mortality occurs. OMW concentrations ranging from 0.01 to 0.1% (v/v) showed no increased mortality and thus were used for the determination of pre-pathological alterations in tissues of mussels. In particular, mussels exposed to either 0.1 or 0.01% (v/v) OMW for 5 days showed significant alterations of stress indices in their tissues. Specifically, decreased neutral red retention (NRR) assay time values, inhibition of acetylcholinesterase (AChE) activity, as well as a significant increase of micronucleus (MN) frequency and DNA damage were detected in haemolymph/haemocytes and gills, compared with values measured in tissues of control mussels. The results of the present study showed that OMW disposal into the marine environment could induce pre-pathological alterations in marine organisms, before severe disturbances, such as disease, mortality, or population changes occur., (2010 Elsevier B.V. All rights reserved.)

In this study, the complex wastewaters/effluents discharged to coastal regions of Turkey and Greece were sampled, and various toxicity tests were applied. The bioassays used included the assessment of the luminescence inhibition of the bacterium Vibrio fisheri , the photosynthesis inhibition ( 14 C uptake rate) and growth inhibition of the algal species Phaeodactylum tricornutum , and the mortality of the crustacean Artemia franciscana and rotifer Brachionus plicatilis . Additionally, “neutral red retention” and “filtration rate” sublethal biomarker techniques were performed by utilizing the mussel species Mytilus galloproviancialis , inhabiting the points of discharges in Turkey. All discharges tested were found to be acutely toxic to P. tricornutum and slightly toxic to V. fisheri, A. franciscana , and B. plicatilis. Test results showed that the 14 C uptake rate toxicity test was the most sensitive one among the other bioassays. The biomarker results showed that the health status of mussels in the coastal areas decreased significantly, indicating the pollution of the receiving waters in Turkey. The conclusions drawn highlight the necessity for an intesive ecotoxicological monitoring scheme that will incorporate the most suitable bioassays and biomarkers to adequately contribute to the upgrading and maintenance of the ecological quality of the coastal waters in Greece and in Turkey. [ABSTRACT FROM AUTHOR]

Satureja pilosa Velen. (Lamiaceae) is a perennial and melliferous aromatic–medicinal subshrub which is range-restricted in adjacent parts of Greece and Bulgaria and locally in Italy, known in Northern Greece as wild oregano ("agriorigani") and traditionally collected from the wild for culinary purposes. Since the ethnopharmacological data and modern biological activities of Satureja spp. suggest promising applications in skin conditions, the present study aimed to investigate the hitherto unknown phenolic content of cultivated S. pilosa and its potential biological activities, focusing mainly on wound-healing and anti-inflammatory effects. An HPLC-PDA-MS-targeted phytochemical investigation, along with NMR, allowed for the isolation and characterization of the main constituents, resulting in 18 compounds. Representative extracts and purified compounds were tested for wound-healing activity on NIH/3T3 fibroblasts. The butanol extract exhibited a significantly higher cell migration rate (73.4%) compared to aqueous (50.6%) and methanolic (49.6%) ones, enhancing the cell migration more rapidly at both concentration levels, whilst rosmarinic acid was the most potent among the isolated compounds, with a migration rate of 64.0% at the concentration level of 10−5 mg/mL, followed by 3,4-dihydrophenyllactic acid (54.7%). Moreover, potential effects on endothelial activation processes were explored, including the leukocyte–endothelial cell interaction during inflammatory processes and the migratory capacity during angiogenic actions, since these processes are commonly associated with skin diseases. Finally, extracts and purified compounds demonstrated weak antibacterial potential against two important pathogens (Staphylococcus aureus and Pseudomonas aeruginosa), suggesting that further investigation is warrented. [ABSTRACT FROM AUTHOR]

Ginsenoside metabolite compound K (CK) is an active metabolite produced by ginsenosides in vivo that has an anti-arthritic effect related to the glucocorticoid receptor (GR). However, the potential mechanisms of CK remain unclear. This study explores the role and potential mechanisms of CK in vivo and in vitro. Adjuvant arthritis (AA) model was induced in Sprague-Dawley (SD) rats; the rats were randomly divided into four groups (n = 10): normal, AA, CK (80 mg/kg), and dexamethasone (Dex) group (1 mg/kg). From day 15, rats were treated with CK (once a day, i.g.) and Dex (once every 3 days, i.p.) for 18 days. To further verify the mechanism of CK, fibroblast-like synoviocytes (FLS) were stimulated by tumour necrosis factor α (TNF-α) to establish an inflammatory model in vitro. CK (80 mg/kg) reduced paw swelling (52%) and arthritis global assessment (31%) compared to that in AA rats. In addition, CK (80 mg/kg) suppressed GLUT1 (38%), HK2 (50%), and PKM2 (56%) levels compared with those in AA FLS. However, the effects of CK (30 μM) on these events were weakened or enhanced after GR knockdown or overexpression in FLS stimulated by TNF-α (30 ng/mL). CK (80 mg/kg) also downregulated the expression of P65 (61%), p-IκB (92%), and HIF-1α (59%). The inhibition of CK on glycolysis and the NF-κB/HIF-1α pathway is potentially mediated through activating GR. These findings provide experimental evidence for elucidating the molecular mechanism of CK in treating rheumatoid arthritis (RA). [ABSTRACT FROM AUTHOR]

Lamiaceae family includes various medicinal and aromatic plants used in cosmetics and pharmaceutical industries. The present study aimed to investigate in vitro the cytotoxic, photoprotective and antioxidant activities of ten Lamiaceae taxa; Melissa officinalis subsp. altissima (Sm.) Arcang., Rosmarinus officinalis L., Salvia officinalis L., Sideritis cypria Post, S. euboea Heldr., S. perfoliata L. subsp. perfoliata, S. scardica Griseb., S. sipylea Boiss., Stachys iva Griseb., and Thymus vulgaris L. The aqueous extract of Salvia officinalis was bio-guided fractionated to obtain the main bioactive metabolites, which were evaluated for the aforementioned effects and their wound-healing potential. In total, five compounds were isolated and identified through NMR spectra, namely salvianic acid A, rosmarinic acid, salvianolic acid K, luteolin-3′-O-β-D-glucuronide and hispidulin-7-O-β-D-glucuronide. All the compounds were photoprotective and non cytotoxic, while no statistically significant oxidative stress reduction was obtained. Regarding the wound-healing potential, salvianolic acid K was the most promising candidate. Overall, this study suggests photoprotective natural agents from various Lamiaceae species, widely found in Greece, and provides a better insight into Salvia officinalis and its bioactive constituents. [ABSTRACT FROM AUTHOR]

Tribulus terrestris L. is an herbaceous plant native to India, widely used as a natural sexual stimulant by traditional medicine in China, India, and Greece, and also for the treatment of various diseases for example low blood pressure, cardiovascular diseases, kidney, and skin diseases, etc. Tribulus extracts are rich in numerous biologically active compounds, such as steroidal saponins, alkaloids, lignan amides, and flavonoids, which are useful for the effective treatment of sexual performance, hormonal and sexual problems. The main goal of our in vitro study was to demonstrate the potential consequences of Tribulus terrestris L. extract on TM3 cell parameters such as cell viability, membrane integrity, lysosomal activity, and steroid hormone secretion. The extract was applied to mice Leydig cells at various concentrations (ranging from 37.5 µg/mL to 600 µg/mL) for 24 h. Based on the obtained data, we may claim statistically significant inhibition of cell viability at 300 µg/mL (P < 0.0001) and 600 µg/mL (P < 0.0001). At the same concentrations, there was a statistically significant decline in both cell membrane integrity (P < 0.05; P < 0,0001) and lysosomal activity (P < 0.05; P < 0.001). Hormone secretion was significantly increased at 200 µg/mL (P < 0.05). To better understand how Tribulus terrestris L. affects the reproductive systems of animals or humans, further research must be done. [ABSTRACT FROM AUTHOR]

The present study aimed to investigate the chemical profile and the protective activity on fibroblasts of two Matricaria species: M. pubescens, which grows wild in Algeria, and M. recutita, which is cultivated in Greece. A comparative phytochemical investigation using High-Performance Liquid Chromatography, coupled with Photodiode Array Detection and Mass Spectrometry (HPLC–PDA–MS) combined with Nuclear Magnetic Resonance (NMR), was performed for the identification of the main constituents in the flowerheads of these medicinal plants. In M. pubescens more than 25 compounds were identified and/or isolated; among them are quercetagenin-3-O-glucopyranoside, reported for the first time in Matricaria sp., and two polyamines previously reported in other Asteraceae species. In M. recutita, which is the officially recognized species in Europe, 19 constituents were identified. To minimize time analysis, the structure elucidation was based on a multi-analytical approach directly on subfractions. Two representative polar extracts from each species were characterized chemically and further screened for their protective effects on 3T3 fibroblasts. The cells were exposed to a mild toxic dose of UVA light (6 J/cm2), in the presence of different concentrations of the extracts. Both M. recutita and M. pubescens extracts were effective. The methanolic extract was the best protective agent at lower concentrations (0.1 to 10 μg/mL), and hydromethanolic was best at higher ones (100–200 μg/mL). M. recutita exhibited the most enhanced cell viability in relation to those not exposed to UV control; it ranged from of 28 to 49% higher viability, depending on the dose, leading to the conclusion that the latter seems to exhibit potent cytoprotective activity and significant regeneration activity. [ABSTRACT FROM AUTHOR]

Purpose: To compare in vitro the attachment and proliferation of human osteoblast-like cells (MG63) on tissue culture plates and guided bone regeneration (GBR) membranes in the absence or presence of nicotine. Materials and Methods: Membrane samples were fixed to wells and the cell number (CN) was counted after 24 hours (attachment assay) or 5 days (proliferation assay). The ratio of cell count (RCC) (CN at 5 days/CN at 24 hours) was calculated. The study had three parts: First, five different resorbable GBR membranes were compared (Resolut Adapt LT [RALT], Biocollagen [BC], Bio-Gide [BG], OsseoGuard [OG], and Demokritos Human Tissue Bank [DEM]). Next, cells were cultured on tissue culture plates with five different concentrations of nicotine. Finally, cells were cultured on the membrane that had demonstrated the highest RCC and CN in part 1 with four different concentrations of nicotine. Results: At 24 hours, BG showed the highest CN and OG showed the lowest CN. At 5 days, BG showed the highest CN. The order of RCC was BG > OG > DEM > RALT ~ BC. At 24 hours, lower nicotine concentrations (0.3 and 3 ?g/mL) showed higher CNs versus the control, whereas CNs for high nicotine concentrations (30 and 300 ?g/mL) were lower than for the control. CN at 5 days and RCC were lowest with 300 ?g/mL nicotine. At 24 hours and 5 days, all differences among wells with membrane were statistically insignificant. Nevertheless, CN at 5 days and RCC were highest with the lowest nicotine concentration (3 ?g/mL) and lowest with high concentrations. Conclusions: Membrane materials influence attachment and proliferation of bone cells and, therefore, could affect the outcomes of GBR. On both tissue culture plates and membranes, there is a tendency toward a biphasic effect of nicotine, with stimulatory effects at low concentrations and inhibitory effects at high concentrations. [ABSTRACT FROM AUTHOR]

Focuses on the importance of war between technologically advanced states as a stimulus to medical progress. Improvement of health in Greece after the enemy occupation in 1944; Mission of British and U.S. governments to promote the wellbeing of liberated peoples; Causes of improvements in health.

Temporary henna tattoos have become increasingly popular as a safe alternative to permanent tattoos among American and European children and teenagers during the summer holidays. Currently, temporary henna tattoos contain not only henna, but also other additives such as para-phemylenediamine (PPD), which is considered to be the chemical agent that most frequently causes skin reactions associated with the use of commercial black henna. In this report, we describe an 11-year-old boy who applied a temporary black henna tattoo on his right arm during the summer holidays in Greece and developed a severe contact dermatitis at the tattoo site with residual hypopigmentation. He had no previous history of contact dermatitis, however he did suffer from seasonal allergic rhinitis and atopic dermatitis. Patch testing revealed a strong reaction to PPD, a substance commonly contained in temporary henna tattoo preparations. Conclusion: Henna tattoos are an increasing problem worldwide since they carry an increased risk of severe skin reactions; therefore we suggest that the use of temporary henna tattoos in children be discouraged. [ABSTRACT FROM AUTHOR]

The cytotoxic effects of flunitrazepam, zipeprol and trihexyphenidyl on cell viability of peripheral blood mononuclear cells from healthy volunteers were studied at concentrations from 10(-2) to 10(-8) M by 2 colorimetric in vitro assays: the neutral red uptake assay and thiazolyl blue tetrazolium bromide assay. All tested drugs of abuse were non-cytotoxic at concentrations lower than 10(-5) M. Possible immunomodulative effects of these substances were evaluated through phytohemagglutinin induced lymphocyte proliferation ([3H]-thymidine DNA incorporation assay) and by a 51Cr release natural killer assay. The 3 drugs studied produced statistically significant immunomodulative alterations on both immunological parameters.