Briarenols W–Z: Chlorine-Containing Polyoxygenated Briaranes from Octocoral Briareum stechei (Kükenthal, 1908)

Briareum stechei is proven to be a rich source of 3,8-cyclized cembranoids (briarane) with a bicyclo[8.4.0] carbon core. In the present study, four previously unreported briaranes, briarenols W–Z (1–4), along with solenolide A (5), briarenolide M (6), briaexcavatolide F (7), and brianolide (8), were isolated and characterized through spectroscopic analysis, and the absolute configuration of 8 was corroborated by a single-crystal x-ray diffraction analysis. Briaranes 2 and 5 were found to induce significant inflammatory activity in lipopolysaccharide (LPS)-induced RAW 264.7 mouse macrophage cells by enhancing the expression of the inducible nitric oxide synthase (iNOS) and cyclooxygenase-2 (COX-2) proteins.     

Antarctic Marine Algae Extracts as a Potential Natural Resource to Protect Epithelial Barrier Integrity

The intestine and skin provide crucial protection against the external environment. Strengthening the epithelial barrier function of these organs is critical for maintaining homeostasis against inflammatory stimuli. Recent studies suggest that polar marine algae are a promising bioactive resource because of their adaptation to extreme environments. To investigate the bioactive properties of polar marine algae on epithelial cells of the intestine and skin, we created extracts of the Antarctic macroalgae Himantothallus grandifolius, Plocamium cartilagineum, Phaeurus antarcticus, and Kallymenia antarctica, analyzed the compound profiles of the extracts using gas chromatography-mass spectrometry, and tested the protective activities of the extracts on human intestinal and keratinocyte cell lines by measuring cell viability and reactive oxygen species scavenging. In addition, we assessed immune responses modulated by the extracts by real-time polymerase chain reaction, and we monitored the barrier-protective activities of the extracts on intestinal and keratinocyte cell lines by measuring transepithelial electrical resistance and fluorescence-labeled dextran flux, respectively. We identified bioactive compounds, including several fatty acids and lipid compounds, in the extracts, and found that the extracts perform antioxidant activities that remove intracellular reactive oxygen species and scavenge specific radicals. Furthermore, the Antarctic marine algae extracts increased cell viability, protected cells against inflammatory stimulation, and increased the barrier integrity of cells damaged by lipopolysaccharide or ultraviolet radiation. These results suggest that Antarctic marine algae have optimized their composition for polar environments, and furthermore, that the bioactive properties of compounds produced by Antarctic marine algae can potentially be used to develop therapeutics to promote the protective barrier function of the intestine and skin.     

Acute haemolysis associated with etomidate-propylene glycol infusion in dogs

Acute haemolysis occurred in medetomidine-atropine premedicated dogs (n=6) after infusion of etomidate in 35% propylene glycol (etomidate-PG). Free plasma haemoglobin concentration was 12.0 +3.5 μg/dl at baseline. After premedication (medetomidine 15 μg/kg, IM; atropine 0.044 mg/kg, IM) values were 14 ± 5.2 and 20 ± 4.8 mg/dl, at 5 and 10 minutes, respectively. Plasma haemoglobin values increased significantly (p±0.05; 121 +24.2 mg/dl) 5 minutes after etomidate-PG loading dose (0.5 mg/kg) and infusion (50μg/kg/min) and remained significantly elevated (127 ± 12.7 to 310.6 ± 69.3 mg/dl) throughout the 60-minute infusion period. Acute haemolysis was also observed in dogs (n=3) that received etomidate-PG infusion alone (2 mg/kg loading dose followed by 110 μg/ kg/ min constant infusion). In addition, fresh dog blood (n=3) was incubated alone or with either 0.9% saline or etomidate-PG in test tubes for 5 minutes and free plasma haemoglobin concentration measured. Free plasma haemoglobin concentrations were 18.3 ± 6.8, 11.7 +4.5 and 1712.0 ± 309.6 mg/dl for blood alone, saline-blood and etomidate-PG-blood, respectively. It was concluded that etomidate-PG caused acute haemolysis in dogs both in vivo and in vitro. The clinical significance of this amount of haemolysis is not clear at this time and thus, requires further study.     

Karyotype evaluation among young horse populations in Poland

Five hundred young horses of the following breeds: Thoroughbred, Silesian, Malopolska, Wielkopolska, Polish Konik, Hutsul, Shetland Pony, Half-bred Anglo-Arabian, Noble Half-bred, Fjord and crosses were cytogenetically investigated. Chromosome preparations obtained after lymphocyte culture were analysed using conventional Giemsa staining and CBG-banding methods. In the case of abnormalities GTG-banding as well as FISH technique were applied. In ten mares different karyotypic abnormalities were diagnosed. One mare showed chromosome chimerism (64,XX/64,XY), eight had sex chromosomal aneuploidy (one in pure line 63,X and seven in mosaic form 63,X/64,XX) and one presented autosomal aneuploidy with mosaicism (64,XX/65,XX,+31). The influence of sex chromosome abnormalities on fertility and the possible utilisation of karyotypic control in any selection programme are discussed.     

1,3-beta-glucan quantification by a fluorescence microassay and analysis of its distribution in foods

The objective of this study was to establish analytical approaches to quantify 1,3-beta-glucan (1,3-beta-G) in foods. Six food categories including legumes, cereals, tubers, vegetables, fruits, and mushrooms and 17 total items were tested. An extraction procedure was designed to prepare food cold-water soluble, hot-water soluble, cold-alkaline soluble, and hot-alkaline soluble fractions. A fluorescence microassay based on aniline blue dye, which bound specifically to 1,3-beta-G, was developed to measure its content in the food fractions. Curdlan was used as standard to construct the 1,3-beta-G calibration curve, and a linear correlation within a 14 microg/mL concentration range was obtained. This microassay displayed selectivities among various 1,3-beta-G species. Biologically active ones such as pachyman and yeast glucan possessed much stronger fluorescent signals than others such as laminarin and barley glucan. Possible fluorescent interference from food proteins was estimated. This assay tolerated up to 50% of bovine serum albumin in 10 microg/mL curdlan. Analysis of the four food fractions showed that besides the well-known lentinan-containing shiitake, popular foods such as celery, chin-chian leaves, carrot, and radish contained nearly 20% 1,3-beta-G in their total sugar. Soybean dry weight contained 0.8% 1,3-beta-G, which was twice the amount compared to shiitake. Snow mushroom dry weight had the highest 1,3-beta-G content, at 2.5%, and was rich in both water (0.67%) and alkaline soluble (1.87%) forms. In conclusion, this dye-binding fluorescence microassay in conjunction with the extraction procedure can be applied in the prescreening of potential foods rich in functional 1,3-beta-G.     

Pseudoalteromone A,a Ubiquinone Derivative from Marine Pseudoalteromonas spp., Suppresses Melanogenesis

An ubiquinone derivative, pseudoalteromone A (1), has been isolated from two marine-derived Pseudoalteromonas spp., APmarine002 and ROA-050, and its anti-melanogenesis activity was investigated. The anti-melanogenic capacity of pseudoalteromone A was demonstrated by assessing the intracellular and extracellular melanin content and cellular tyrosinase activity in the B16 cell line, Melan-a mouse melanocyte cell line, and MNT-1 human malignant melanoma cell line. Treatment with pseudoalteromone A (40 μg/mL) for 72 h reduced α-melanocyte-stimulating hormone (α-MSH)-induced intracellular melanin production by up to 44.68% in B16 cells and 38.24% in MNT-1 cells. Notably, pseudoalteromone A induced a concentration-dependent reduction in cellular tyrosinase activity in B16 cell, and Western blot analyses showed that this inhibitory activity was associated with a significant decrease in protein levels of tyrosinase and tyrosinase-related protein 1 (Tyrp-1), suggesting that pseudoalteromone A exerts its anti-melanogenesis activity through effects on melanogenic genes. We further evaluated the skin-whitening effect of pseudoalteromone A in the three-dimensional (3D) pigmented-epidermis model, MelanoDerm, and visualized the 3D distribution of melanin by two-photon excited fluorescence imaging in this human skin equivalent. Collectively, our findings suggest that pseudoalteromone A inhibits tyrosinase activity and expression and that this accounts for its anti-melanogenic effects in melanocytes.     

Quantitative determination of 12-hydroxyeicosatetraenoic acids by chiral liquid chromatography tandem mass spectrometry in a murine atopic dermatitis model

Atopic dermatitis, one of the most important skin diseases, is characterized by both skin barrier impairment and immunological abnormalities. Although several studies have demonstrated the significant relationship between atopic dermatitis and immunological abnormalities, the role of hydroxyeicosatetraenoic acids (HETE) in atopic dermatitis remains unknown. To develop chiral methods for characterization of 12-HETE enantiomers in a 1-chloro-2,4-dinitrochlorobenzene (DNCB)-induced atopic dermatitis mouse model and evaluate the effects of 12-HETE on atopic dermatitis, BALB/c mice were treated with either DNCB or acetone/olive oil (AOO) to induce atopic dermatitis, after which 12(R)- and 12(S)-HETEs in the plasma, skin, spleen, and lymph nodes were quantified by chiral liquid chromatography-tandem mass spectrometry. 12(R)- and 12(S)-HETEs in biological samples of DNCB-induced atopic dermatitis mice increased significantly compared with the AOO group, reflecting the involvement of 12(R)- and 12(S)-HETEs in atopic dermatitis. These findings indicate that 12(R)- and 12(S)-HETEs could be a useful guide for understanding the pathogenesis of atopic dermatitis.     

广州地区十字花科蔬菜花叶病第三类病原病毒的鉴定

广州地区十字花科蔬菜花叶病第三类病原病毒的物理性质与TMV基本相似:失毒温度86—90℃之間,寄主体外存活期限37个月以上,稀释終点1:10000—50000之間。本病毒易由液汁接触传染,可由斜纹夜盜蛾、菜粉蝶和甘蓝夜蛾的2—3龄幼虫、普通紅蜘蛛以及中国菟絲子传染。但不能由桃蚜、蘿卜蚜、黃条跳(虫甲)成虫以及感病蔬菜的种子传染。在含新鮮病菜組織的土壤中移植蔬菜,可引起部分蔬菜感染发病。本病毒的寄生范围較广。在供試植物20科49屬93个种和品种中,能侵染十字花科、茄科、菊科、藜科、莧科、車前草科和石竹科植物等7科18屬54种和品种。交互保护試驗結果,TMV(毒株23号)对本病毒只有小部分的保护作用。但根据周家熾、田波等試驗結果認为本病毒与TMV有一定的血清学关系,因此暫把此病毒列入TMV类羣中,作为一个远缘的毒株。     

Protoporphyrinogen-IX Oxidase Inhibitors: Bioactivation of Thiadiazolidines

The bioactivation of thiadiazolidine-type peroxidizing compounds was examined with thiosemicarbazides as model intermediates. Peroxidizing activities of three sets of thiadiazolidines (5-arylimino-3,4-tetramethylene-1,3,4-thiadiazolidines), thiosemicarbazides (4-aryl-1-ethylthiocarbonyl-1,2-tetramethylenethiosemicarbazides), and triazolidines (4-aryl-1,2-tetramethylene-1,2,4-triazolidines) were assayed for protopor- phyrinogen-IX oxidase (Protox) inhibition with Protox isolated from corn etioplasts and for phytotoxic parameters of growth, chlorophyll content, and ethane evolution obtained with Scenedesmus acutus cells. Protox inhibitory activity of thiosemicarbazides was intermediate between that of thiadiazolidines and triazolidines. Phytotoxic parameters of thiosemicarbazides obtained from S. acutus cells were quite identical to those of triazolidines, although phytotoxic parameters of thiadiazolidines exhibited a slightly different pattern. Phytotoxic activities of 5-(4-bromophenyl)-3,4-tetramethylene-1,3,4-thiadiazolidin-2-one (thiadiazolidine 1) and 5-(4-chloro-2-fluoro-5-propargyloxyphenyl)-3,4-tetramethylene-1,3,4-thiadiazolidin-2-one (thiadiazolidine 4) were similar to those of the corresponding triazolidines, but phytotoxicities exhibited by 5-(4-(4-chlorobenzyloxy)phenyl)-3,4-tetramethylene-1,3,4-thiadiazolidin-2-one (thiadiazolidine 7) were 10 times less active with S. acutus than those of the corresponding triazolidine. Three thiosemicarbazides exhibited rapid conversion into triazolidines both in buffer at pH 7 (ca. 25–50%, 5 min incubation) and in Scenedesmus cultures (ca. 75%, 5 h incubation). Thiadiazolidines 1 and 4 were converted into corresponding triazolidines (ca. 70%) in S. acutus but were converted in buffer only in the presence of glutathione S-transferase plus ethylmercaptan, although thiadiazolidine 7 was not converted in the Scnedesmus culture. Apparently, the conversion step from thiadiazolidine to the thiosemicarbazide intermediate is enzymatic and the thiosemicarbazide intermediate to triazolidine is a spontaneous, nonenzymatic step.