Isolation and Characteristic of Overlong-Chain Unsaturated Aldehydes from the Freshwater Sponge <Emphasis Type="Italic">Lubomirskia baicalensis</Emphasis>

Several unusual overlong-chain unsaturated aldehydes (22 : 1, 22 : 2, 23 : 1, 24 : 1, 24 : 2, and 25 : 2) were found in total lipids of the endemic sponge Lubomirskia baicalensis from Baikal Lake. Tetracos-17-enal was identified as the major aldehyde of the mixture using GC-MS and ¹H and ¹³C NMR spectroscopy. A procedure for the isolation of total overlong-chain aldehydes was suggested. We think that the overlong-chain aldehydes defend the sponge from fouling and predators.     

The Composition of fatty acids and aldehydes of the marine bryozoans <Emphasis Type="Italic">Berenicea meandrina</Emphasis> and <Emphasis Type="Italic">Dendrobeania flustroides</Emphasis> (Bryozoa: Gymnolaemata)

This study examines the composition of lipids, fatty acids, and fatty aldehydes in two marine bryozoan species, Berenicea meandrina and Dendrobeania flustroides, from the Sea of Okhotsk. The share of neutral lipids was up to 57.3% in D. flustroides and 54.9% in B. meandrina; the share of polar lipids was 33.2 and 40.4%, respectively. In all, 57 fatty acids (FA) and 9 aldehydes were identified in total lipids. The main FAs were 16:0, 18:0, 22:6n-3, and 20:5n-3. The content of branched saturated FA in bryozoans was on the average 6.4%. Three isomers of 16:1 (n-9, n-7, and n-5), five isomers of 18:1 (n-13, n-11, n-9, n-7, and n-5), four isomers of 20:1 (n-13, n-11, n-9, and n-7), as well as 22:1n-9 and 22:1n-13 were found; the presence of 7-methyl-6-hexadienoic acid (on the average, 3.0% of total FAs) was demonstrated. Non-methylene-inter-rupted FAs contributed 8.9 and 1.6% of the total FAs in D. flustroides and B. meandrina, respectively, and were identified as 20:2(5,11), 20:2(7,13), 20:3(5,11,14), 22:2(7,13), and 22:2(7,15). In B. meandrina, minor amounts of 24:0, 24:1, 25:0, 26:0, 24:4n-3, 26:3(5,9,19), and 28:3(5,9,19) were found, suggesting sponge biofouling on some bryozoan colonies. Aldehydes (branched saturated and unsaturated C_16–19 homologues) did not exceed 10.3 and 1.9% of the total FAs in D. flustroides and B. meandrina, respectively. The presence of the FA markers that are characteristic of microalgae, protozoans, and detritus in bryozoan lipids agrees well with data on polytrophic feeding of these bryozoans.     

Ophirasterol, a new C31 sterol from the marine sponge Topsentia ophiraphidites

Analysis of the sterol fraction obtained from the Colombian Caribbean sponge Topsentia ophiraphidites revealed that this sponge is a rich source of C30 and C31 sterols. Among them, a new C31 sterol, named ophirasterol, was isolated, and its structure was established as (22E,24R)-24-(1-buten-2-yl)cholesta-5,22-dien-3beta-ol (1) by spectral means and comparison with synthetic C-24 epimers with known configuration. Other isolated C30 and C31 sterols were the known 24-ethyl-24-methyl-22-dehydrocholesterol (2), 24-isopropyl-22-dehydrocholesterol (3), 24-isopropylcholesterol (4), 24-ethyl-24-methylcholesterol (5), 24-isopropenyl-25-methyl-22-dehydrocholesterol (6) and 24-isopropenyl-25-methylcholesterol (7), and 24-isopropenyl-22-dehydrocholesterol (8).     

Three new 7-keto sterols from the Mediterranean sponge Clathrina clathrus

The CHCl3 extract of the Mediterranean sponge Clathrina clathrus was shown to contain three new sterols: (22 E)-3 beta-hydroxycholesta-5,8,22-trien-7-one (1), (22 E, 24 xi)-3 beta-hydroxy-24-methylcholesta-5,8,22-trien-7-one (2), and (22 E, 24 xi)-3 beta-hydroxy-24-ethylcholesta-5,8,22-trien-7-one (3), whose structures were elucidated on the basis of physico-chemical evidence.     

Minor and trace sterols in marine invertebrates 47. A re-investigation of the 19-nor stanols isolated from the sponge Axinella polypoides

The aim of this research was to establish the true composition of the 19-nor stanols isolated from the sponge Axinella polypoides and to determine accurate stereochemistry for each 19-nor stanol isolated. The following new 19-nor stanols were collected from this sponge: (i) (22E,24S)-24-methyl-19,27-bisnor-5 alpha-cholest-22-en-3 beta-ol, (ii) (22R,23R)-22,23-methylene-19-nor-5 alpha-cholestan-3 beta-ol, (iii) (24 xi)-24-propyl-19-nor-5 alpha-cholestan-3 beta-ol and (iv) (23R,24R)-23,24-dimethyl-19-nor-5 alpha-cholestan-3 beta-ol. The general structure and stereochemistry of all fifteen 19-nor stanols were established by analysis of the MS and H-NMR (300 MHz, CDCl3) data measured for each compound. The relative percentage of 19-nor stanols having delta 22 double bonds should be sufficient to suggest that this sponge could be a potential source of starting material for the partial synthesis of certain oral contraceptives, which also have a 19-nor steroid nucleus.     

Isolation of 2-methyl branched unsaturated very long fatty acids from marine sponge Halichondria panicea and identification of them by GC-MS and NMR

In order to study the biomarker fatty acids of symbionts in the marine sponge Halichondria panicea, purification and structural identification of two new 2-methyl branched monoenoic very long fatty acids (2-Me-24:1 n-7 and 2-Me-26:1 n-9) were performed for the first time. These acids amounted to 7.1% of total sponge FAs, but our attempts to determine their structures by one-step GC-MS analysis were unsuccessful because of low yields of the correspondent N-acyl pyrrolidide derivatives. Silver-ion thin-layer chromatography isolated enriched fractions of monoenoic fatty acids extracted from the sponge. Further purification of unknown fatty acid methyl esters was carried out by reversed-phase high-performance liquid chromatography. Determination of the chain length, degree and position of unsaturations was achieved by gas chromatography-mass spectrometry on methyl esters and dimethyldisulfide adducts. Structures, position of methyl substitution, and double bonds cis isomery were confirmed by 1H nuclear magnetic resonance.     

Fatty acids from the sponge Halichondria panicea from the Sea of Japan

The fatty acid (FA) composition of total lipids isolated from the marine sponge Halichondria panicea inhabiting Peter the Great Bay of Sea of Japan was studied. GC and GC-MS techniques helped identify 63 FAs, with the main attention being paid to FAs with 14-22 carbon atoms. 4, 8, 12-Trimethyl-13:0 FA was for the first time identified as the main saturated FA along with the branched FAs br-25:1, br-27:1, and br-27:2. The contents of arachidonic, eicosapentaenoic, docosapentaenoic, and the major demospongic acids [26:3(5, 9, 19), 26:3(5, 9, 17), 27:3(5, 9, 20), and 28:3(5, 9, 21)] considerably differed from those previously found for H. panicea, which may be due to seasonal changes in the species composition of organisms consumed by the sponge.     

Stimulus-response coupling in marine sponge cell aggregation: lipid metabolism and the function of exogenously added arachidonic and docosahexaenoic acids

Cells of the marine sponge, Microciona prolifera, the most ancient of the animal cells which clump on recognition, resemble neutrophils and platelets in undergoing stimulus-response coupling when exposed to Ca2+ ionophores and phorbol esters. We have studied lipid content and remodelling in sponge cells by thin-layer, gas-liquid, and high-performance liquid chromatography (HPLC) analyses supplemented by ultraviolet and mass spectroscopy. Phosphatidylcholine (PC) (35.6%), phosphatidylethanolamine (PE) (27.4%) and phosphatidylserine (PS) (21.4%) constituted the bulk of phospholipids detected. The major fatty acids were all polyenoic; 22:6 (22%), 26:2 (17%) and 26:3 (15%). Arachidonic acid (20:4), present as 2.7% of total phospholipid, and docosahexanoic acid (22:6) were found to elicit aggregation of sponge cells when added (10 microM) in synergy with ionomycin (1 microM), resembling in their effects those of phorbol esters (but not phorbol) and 1-oleyl-2-acetylglycerol (OAG). Moreover, 20:4 and 22:6, as well as phorbol ester and OAG, overcame the block to aggregation imposed by colchicine and vinblastine. Kinetic studies of lipid remodelling showed that aggregating cells diverted [14C]22:6 or [14C]20:4 from triacylglycerol into diacylglycerol and phospholipids; appearance of label in phosphatidic acid and phosphatidylinositol (PI) anteceded labeling of phosphatidylcholine. In unstimulated cells, [14C]22:6 was rapidly incorporated into phosphatidylcholine with little accumulation in phosphatidate. Although 22:6 and 20:4 resembled OAG and phorbol esters in overcoming the effects of colchicine and vinblastine (which had no effects on overall lipid metabolism), they did not reverse the block to aggregation of nordihydroguaiaretic acid (NDGA) (which inhibited lipid metabolism). Under none of these circumstances was 22:6 or 20:4 converted to cyclooxygenase or lipoxygenase products in the course of aggregation: all labeled acyl groups remained present as unmodified fatty acids on alkaline hydrolysis. These data not only extend the observations of Muller et al. (J. Biol. Chem. 262 (1987) 9850-9858) on the role of phosphoinositides and C kinase in marine sponge cell aggregation, but also demonstrate that sponges form diacylglycerols in the process. We suggest that exogenous 22:6 and 20:4 (like phorbol esters or OAG) can substitute for endogenous diacylglycerol in the activation of protein kinase C.     

Novel triterpene glycosides from the sponge Ulosa sp

New triterpene glycosides, ulososides C, (20S,22S,23R,24S)-3 beta,22, 23-trihydroxy-3-O-(beta-D-glucopyranosyl)-32-nor-24-methyllanost- 8(9)-ene-30-oic acid, D, (20S,22S,23R,24S)-3 beta,22, 23-trihydroxy-3-O-(beta-D-N-acetyl-glucosaminopyranosyl)-32-nor- 24-methyllanost-8(9)-ene-30-oic acid, and E, (20S,22S,23R,24S)-3 beta,22, 23-trihydroxy-3-O-(beta-D-glucuronopyranosyl-(1-->2)-alpha-D- arabinopyranosyl-32-nor-24-methyllanost-8(9)-ene-30-oic acid, were isolated from an Ulosa sp. sponge. Their structures were determined by spectral methods and chemical transformations. Specific features of their structures are discussed.     

Steroids in Porifera. II. Steroid derivatives from two sponges of the family Halichondriidae. Sokotrasterol sulfate, a marine steroid with a new pattern of side chain alkylation

A trisulfated derivative of 24,25,26,26-tetramethyl-5 alpha-cholest-23E-ene-2 alpha, 3 beta, 6 alpha-triol (sokotrasterol sulfate) has been isolated from the sponge Halichondriidae gen. sp., collected near Sokotra Island (Arabian Sea), and its structure has been elucidated. The side chain of the new steroid involves a "normal" alkylation at C-24 and the unprecedented addition of two extra methyl groups at C-26 and one extra methyl group at C-25. A free sterol fraction contained only 24-isopropyl-5-cholesten-3 beta-ol and 24-isopropyl-5, 22E-cholestadien-3 beta-ol. 24-Isopropyl-5, 22E-cholestadien-3 beta-ol as sole monohydroxy sterol and halistanol sulfate as major polyhydroxylated steroid derivative have been detected in Halichondria sp., a Madagascar sponge.     

Glucosylceramide having a novel tri-unsaturated long-chain base from the spermatozoa of the starfish, Asterias amurensis

Glucosylceramide (Glc beta 1-1Cer) was isolated from the spermatozoa of the starfish, Asterias amurensis. The long-chain bases of the glycolipid consisted of dihydroxy (d18:2, d18:3, d19:3, and d22:2), and trihydroxy (t22:1) types. Long-chain aldehydes derived from them were analyzed mainly by proton nuclear-magnetic resonance to determine the detailed structures. Two of the tri-unsaturated bases were identified as (4E,8E,10E)-2-amino-4,8,10-octadecatriene-1,3-di ol (d18:3) and (4E,8E,10E)-2-amino-9-methyl-4,8,10-octadecatriene+ ++-1,3-diol (d19:3), which is a novel base. Both d22:2 and t22:1 had a cis double bond at the C9 or C13 position. All fatty acids were 2-hydroxylated (C14-C25): Most of them were saturated and unbranched. About 10% was mono-unsaturated and unbranched (C22-C25), while saturated but branched (iso- and anteiso-types) C15-C18 acids were found as minor components. The main fatty acids, which summed up to more than 93% of the fatty acids in the glucosylceramide, were n-14h:0, n-15h:0, n-16h:0, n-17h:0, n-18h:0, and n-24h:1.     

Effects of time of PMSG and fixed-time GnRH injections on estrus incidence and fertility in physiologically different ewes pre-treated with progestogen-impregnated vaginal sponge during the nonbreeding season

A 2 × 2 factorial study for hormonal treatment was designed in 85 seasonally anestrous ewes with physiologically different status (nonparous, dry, and postpartum). All ewes were pre-treated with 60 mg of 6-methyl-17-acetoxy-progesterone (MAP) vaginal sponge for nine days and divided into four groups: Group I (22 ewes) — an i. m. injection of 600 i.u. pregnant mare's serum gonadotropin (PMSG) at the sponge removal (Day 0) and a single i.m. injection of 100 ug synthetic gonadotropin releasing hormone (GnRH) at 36 h after the sponge removal; Group II (21 ewes) — a PMSG injection at Day 0 and a saline injection at 36 h after the sponge removal; Group III (21 ewes) — a PMSG injection two days before the sponge removal and the GnRH injection at 24 h after the sponge removal; and Group IV (21 ewes) — a PMSG injection at Day -2 and a saline injection at 24 h after the sponge removal. The treated ewes were allowed to mate once with rams for five days after treatment. Estrus incidence and lambing rates were low (69.4% and 27.1%, respectively), probably due to the mating system and poor body condition of ewes used in the study. No effect of PMSG injection two days before with-drawal of MAP sponge and the fixed-time GnRH injection were observed in estrus incidence, fertility, and prolificacy. The present study indicates that the physiological status of ewes combined with management systems including feeding and mating would be important for out-of-season breeding with hormonal treatment.     

Fatty Acids from the Sea of Japan Sponge Halichondria panicea

The fatty acid (FA) composition of total lipids isolated from the marine sponge Halichondria panicea inhabiting Peter the Great Bay of the Sea of Japan was studied. GC and GC-MS techniques were used in identification of 63 FAs, with the main attention being paid to FAs with 14–22 carbon atoms. 4,8,12-Trimethyl-13:0 FA was for the first time identified as a main saturated FA, along with the branched FAs br-25:1, br-27:1, and br-27:2. The contents of arachidonic, eicosapentaenoic, docosapentaenoic, and the major demospongic acids [26:3(5,9,19), 26:3(5,9,17), 27:3(5,9,20), and 28:3(5,9,21)] considerably differed from those previously found for H. panicea, which may be due to seasonal changes in the species composition of organisms consumed by the sponge.     

Isolation and Structure Elucidation of Epipolasterol and 22,23-Dihydroepipolasterol from the Marine Sponge Epipolasis sp.

Two new sterols, epipolasterol and 22(23)-dihydroepipolasterol, have been isolated from the marine sponge Epipolasis sp. These are unusual metabolites as they both contain a t-butyl group in the sterol side chain. In addition, the presence of two degrees of unsaturation in the side chain of epipolasterol is rare. The known sterol, 22-dehydro-24-isopropylcholesterol was also found in this sponge.     

泰绫及明胶海绵在乳腺癌手术中的应用

目的:观察可吸收性止血纱布(商品名:泰绫)及明胶海绵用于乳腺癌改良根治术的止血效果及其对引流量及拔管时间的影响。方法:将180例准备接受乳腺癌改良根治术的患者,随机分为泰绫组、明胶海绵组和对照组,每组各60例,术后分别给予泰绫、明胶海绵及空白对照处理,记录和比较两组术后第1、2、3天的引流量、引流液红细胞计数、总引流量及拔管时间。结果:与对照组比较,泰绫组、明胶海绵组术后1、2、3天的引流量均显著减少、引流液红细胞计数、总引流量均明显减少且拔管时间显著缩短,差别均有统计学意义(P0.05)。与明胶海绵组相比,泰绫组术后1、2、3天引流量、引流液红细胞计数、总引流量及拔管时间亦显著减少或缩短,差别均有统计学意义(P0.05)。结论:泰绫及明胶海绵在乳腺癌改良根治术后有较好的止血效果,减少了术后出血、渗血、皮下积液,且泰绫较明胶海绵效果更优。     

Superovulation and recovery of zygotes from Nubian and Alpine dairy goats

Thirty-five purebred dairy goats (18 Alpines and 17 Nubians) were subjected to a superovulating hormone program consisting of an 11-d 6alpha-methyl-17alpha-acetoxy-progesterone; (MAP; 60 mg) intravaginal sponge treatment; 125 ug i.m. injections of the prostaglandin F(2alpha) analogue cloprostenol on d 1 and 9 of vaginal sponge treatment; and a 3-d, twice-a-day injection of 2.5 mg of pituitary follicle stimulating hormone (FSH-P) i.m. starting at day 9. Vaginal sponges were pulled the morning of day 11 at the time of the fifth FSH-P injection. Of 40 initiated superovulatory cycles, 33 does (10 Alpines and 23 Nubians) responded with an average of 17.7 (range 1 to 29) ovulations. There was no significant difference between the breeds with respect to corpora lutea (CLs) plus follicles ovarian response. A significantly greater (P< 0.05) number of Nubian does were in estrus and mated by 36 h after MAP sponge removal. All does that responded to treatment had done so within 72 h of sponge removal. Of the seven (17.5%) does that showed no estrous response to hormone treatment, six were Alpines (P < 0.01). Six goats (two Alpines and four Nubians) were subjected to a second hormone treatment cycle after a 45-d rest. Five of six does responded to a second hormone treatment cycle with four of five responding with a lower total ovarian response. The interval from sponge removal to mating did not affect the stage or quality of eggs harvested. Rather, the interval from mating to surgical flushing determined the stage of egg development. All animals examined from 24 to 32 h after initial mating had not ovulated. By 50 h, 20 of 22 does had ovulated. A total of 242 ovulated eggs (63%) was harvested, of which 199 (82%) were fertilized. Day 7 flushings yielded 36 eggs (67%), of which 28 (78%) were fertilized. This rate of superovulation, fertilization, and embryo recovery lends credibility to this technique in its ultimate objective of rapidly increasing the number of offspring from superior animals.     

Silicon deprivation decreases collagen formation in wounds and bone,and ornithine transaminase enzyme activity in liver

We have shown that silicon (Si) deprivation decreases the collagen concentration in bone of 9-wk-old rats. Finding that Si deprivation also affects collagen at different stages in bone development, collagen-forming enzymes, or collagen deposition in other tissues would have implications that Si is important for both wound healing and bone formation. Therefore, 42 rats in experiment 1 and 24 rats in experiment 2 were fed a basal diet containing 2 or 2.6 μg Si/g, respectively, based on ground corn and casein, and supplemented with either 0 or 10 μg Si/g as sodium metasilicate. At 3 wk, the femur was removed from 18 of the 42 rats in experiment 1 for hydroxyproline analysis. A polyvinyl sponge was implanted beneath the skin of the upper back of each of the 24 remaining rats. Sixteen hours before termination and 2 wk after the sponge had been implanted, each rat was given an oral dose of ¹⁴C-proline (1.8 μCi/100 g body wt). The total amount of hydroxyproline was significantly lower in the tibia and sponges taken from Si-deficient animals than Si-supplemented rats. The disintegrations per minute of ¹⁴C-proline were significantly higher in sponge extracts from Si-deficient rats than Si-supplemented rats. Additional evidence of aberrations in proline metabolism with Si deprivation was that liver ornithine aminotransferase was significantly decreased in Si-deprived animals in experiment 2. Findings of an increased accumulation of ¹⁴C-proline and decreased total hydroxyproline in implanted sponges and decreased activity of a key enzyme in proline synthesis (liver ornithine aminotransferase) in Si-deprived animals indicates an aberration in the formation of collagen from proline in sites other than bone that is corrected by Si. This suggests that Si is a nutrient of concern in wound healing as well as bone formation. The U.S. Department of Agriculture, Agricultural Research Service, Northern Plains Area is an equal opportunity/affirmative action employer, and all agency services are available without discrimination. Mention of a trademark or proprietary product does not constitute a guarantee or warranty of the product by the US Department of Agriculture and does not imply its approval to the exclusion of other products that may be suitable.     

The effects of different PMSG doses on estrus behavior and pregnancy rate in Angora goats

Artificial insemination protocols depend on efficient behavioral estrus detection and insemination time in Angora goat. Therefore, we aim to determine the accuracy of an estrus scoring system in Angora goats with different PMSG doses during the breeding season. Does (n: 260) were randomly divided into three groups: group-1 (n: 93), group-2 (n: 85) and group-3 (n: 82). All animals received an intravaginal sponge on day 0 for 11 days, and on the day of sponge insertion 150 μg prostaglandin F2Α was administered. Pregnant mare’s serum gonadotropin was injected 300, 400 and 500 IU intramuscularly 24 h before sponge removal to groups 1, 2 and 3, respectively. Estrus signs were detected with a teaser buck, 24 h after sponge removal according to a visual scoring system. Artificial insemination was performed with 0.25 ml fresh diluted semen at 43 to 45 h after sponge removal. Differences were observed within PMSG groups in terms of standing, tail wagging, courtship behavior, vaginal discharge and vaginal hyperemia (P<0.001). Nevertheless, the most accurate indicators of estrus that result in pregnancy were tail wagging and courtship behavior followed by standing estrus (P<0.05). According to the results obtained, 300 IU PMSG dose is sufficient, both to inseminate at a fixed time (43 to 45 h after sponge removal) and to record the estrus behavior by teaser male 24 h after sponge removal. Higher PMSG doses (400 to 500 IU) altered the timing of ovulation; specifically, 500 IU dose shortened the duration of estrus behaviors. In conclusion, even though the different doses of PMSG displayed similar effects on estrus synchronization and pregnancy rates, we concluded that tail wagging, courtship behavior and standing heat are the most reliable estrus signs for artificial insemination in Angora goat.     

Analyses of Pneumocystis Fatty Acids

The major ester-linked fatty acids of the total lipids extracted from Pneumocystis carinii isolated from the lungs of corticosteroid-trcaicd rats were 16:0. 18:0, 18:1, 18:2 and 20:4. Others detected included 14:0, 16:1 and 22:4. The major sphingolipid fatty acids were 16:0, 18:0,22:0,24:0 and 24:1; others included 14:0, 18:1, 20:0, 23:0, 24:2 and 26:0. The total lipid fatty acid compositions of preparations from appropriate lung controls were similar to those of the organism.     

Minor and trace sterols in marine invertebrates XVII. 1 (24R)-24,26-dimethylcholesta-5,26-dien-3β-01, a new sterol from the sponge Petrosia ficiformis

More than thirty monohydroxy sterols were detected in the marine sponge $\text{Petrosia ficiformis}$, ranging from the ubiquitous 24-norcholesta-5, 22-dien-3β-o1 to the C³⁰ sterol, 24-propylidenecholesterol. A new minor sterol was isolated and shown by spectral analysis and comparison with a synthetic sample to be (24R)-24,26-dimethy1cho1esta-5,26-dien-3β-o1 [26(29)-dehydroap1ysterol] ($\text{12}$).