The effect of mating on female reproduction across hermaphroditic freshwater snails

Sexual conflicts often arise between mating partners because each sex tries to maximize its own reproductive success. One major male strategy to influence a partner's resource allocation is the transfer of accessory gland proteins. This has been shown to occur in simultaneous hermaphrodites as well as in organisms with separate sexes. Although accessory gland proteins affect the investment of resources in both male and female function, we here specifically focus on female investment. In the great pond snail, Lymnaea stagnalis, previous studies found that the accessory gland protein ovipostatin reduced female fecundity by suppressing egg laying in the partner in the short term (days). To investigate whether this reduction in egg laying is a commonly found effect of mating in freshwater snails, we compared egg output for evidence of suppression in isolated and paired snails of eight pulmonate species. Furthermore, we determined whether the suppression of egg laying caused a shift in resource allocation to the eggs. We found that in five of the eight species egg laying was suppressed, with fewer and lighter egg masses being laid when they had access to a mating partner. In mated pairs of L. stagnalis and Biomphalaria alexandrina, allocation of resources to the eggs was altered in opposite ways: individuals of L. stagnalis laid fewer but larger and heavier eggs; individuals of B. alexandrina laid smaller and lighter eggs, with no change in egg numbers. Such changes in the female function are most likely the result of combined effects of receiving accessory gland proteins, and the cost of mating in both male and female roles. Thus, effects of the maternal environment, including the receipt of accessory gland proteins, on offspring investment are not restricted to species with separate sexes.     

A green and effective method for the determination of metalaxyl enantiomers in tobacco and soil by supercritical fluid chromatography–tandem mass spectrometry

We reported a new methodology for the stereoselective determination of metalaxyl enantiomers in tobacco and soil. The QuEChERS (quick, easy, cheap, effective, rugged, and safe) method was used for the extraction and clean-up of the tobacco and soil samples. Separation of the metalaxyl enantiomers was performed on an ACQUITY UPC2 Trefoil CEL1 chiral column coupled with supercritical fluid chromatography with tandem mass spectrometry (SFC-MS/MS), and the run time was only 5 minutes. Under the optimized conditions, the recoveries for the enantiomers were between 78.2% and 93.3% with intraday relative standard deviations (RSDs) ranging from 1.1% to 5.4%. The limit of detection (LOD) for the enantiomers in tobacco and soil varied from 0.005 to 0.007 mg/kg, and the limit of quantitation (LOQ) ranged from 0.017 to 0.020 mg/kg. In this method, only a small amount of methanol was consumed to obtain a rapid stereoselective separation. This proposed method showed good accuracy and precision and might be suitable for fast enantioselective determination of metalaxyl in food and environmental samples. The developed method was further validated by application to the analysis of authentic samples.     

Chiral separation of underivatized amino acids in supercritical fluid chromatography with chiral crown ether derived column

The supercritical fluid chromatographic separation of underivatized amino acids was explored using immobilized chiral crown ether column CROWNPAK CR-I (+) and mass spectrometric detection. The type of modifier, acidic additives, and the role of water were investigated. Enantioseparation was achieved for all 18 amino acids investigated with short retention times (less than 3 minutes) and average resolution of greater than 5.0. Analysis of enantiomerically pure standards demonstrated the D enantiomer eluted first for all amino acids using a CROWNPAK CR-I (+) column.     

Active interfacial dynamic transport of fluid in a network of fibrous connective tissues throughout the whole body

Fluid in interstitial spaces accounts for ~20% of an adult body weight and flows diffusively for a short range. Does it circulate around the body like vascular circulations? This bold conjecture has been debated for decades. As a conventional physiological concept, interstitial space is a micron‐sized space between cells and vasculature. Fluid in interstitial spaces is thought to be entrapped within interstitial matrix. However, our serial data have further defined a second space in interstitium that is a nanosized interfacial transport zone on a solid surface. Within this fine space, fluid along a solid fibre can be transported under a driving power and identically, interstitial fluid transport can be visualized by tracking the oriented fibres. Since 2006, our data from volunteers and cadavers have revealed a long‐distance extravascular pathway for interstitial fluid flow, comprising at least four types of anatomic distributions. The framework of each extravascular pathway contains the longitudinally assembled and oriented fibres, working as a fibrorail for fluid flow. Interestingly, our data showed that the movement of fluid in a fibrous pathway is in response to a dynamic driving source and named as dynamotaxis. By analysis of previous studies and our experimental results, a hypothesis of interstitial fluid circulatory system is proposed.     

经皮穿刺抽取大鼠脑脊液方法的改良

目的: 改进传统经皮穿刺抽取脑脊液的方法,提高采集大鼠脑脊液动物存活率。方法: 90只SD大鼠随机分为对照组、传统穿刺组和改良穿刺组(每组30只)。对照组不进行穿刺。传统穿刺组按反握抓持法进行穿刺取样。改良穿刺组采用左手固定大鼠头部,将其抬高至约135°,右手正握改良1 ml注射器,以枕骨隆突和第一颈椎为标志定位枕骨大孔,在枕骨大孔中心偏下平行进针,抽取脑脊液。分别于第1、4、7日抽取大鼠脑脊液,每次抽取记录取样时间、采集量、脑脊液性状,抽取后3 d观察大鼠存活情况,选取存活大鼠进行重复采样。结果: 传统经皮穿刺取样法采样成功率为63.33%,三次重复采样动物存活率为10%;改良经皮穿刺取样法采样成功率为86.67%,三次重复采样动物存活率为50%,相比传统采样有显著升高(P＜0.01)。结论: 改良经皮穿刺取样法操作简单,耗时少,成功率高。操作无需定位设备,单人可以完成。动物反复取样存活率高,方便进行脑脊液重复采样。     

乙醇对瘤胃液接种稻秸的体外发酵产物及细菌群落结构的影响

旨在研究乙醇对山羊瘤胃液与水稻秸秆厌氧共培养的影响。利用频繁传代的体外发酵技术和高通量测序方法,分析了短链脂肪酸(SCFA)产量和细菌群落的变化。结果表明,经体外培养传代8次的稻秸发酵液的总短链脂肪酸产量显著高于瘤胃液(P<0.01);与未添加乙醇的稻秸发酵液相比,添加乙醇显著提高了乙酸、戊酸和己酸的比例,降低了丙酸和丁酸的比例(P<0.01),总SCFA产量及异丁酸和异戊酸比例无显著差异。与瘤胃液相比,稻秸发酵液的拟杆菌门(Bacteroidetes)相对丰度下降,厚壁菌门(Firmicutes)相对丰度升高(P<0.05),且添加乙醇显著提高了厚壁菌门和放线菌门(Actinobacteria)的相对丰度(P<0.05);添加乙醇使双歧杆菌属(Bifidobacterium)、未定性的毛螺菌属(unidentified Lachnospiraceae)、产琥珀酸菌属(Succiniclasticum)、脱硫弧菌属(Desulfovibrio)和未定性的梭菌属(unidentified Clostridiales)的相对丰度显著升高(P<0.05)。乙醇使稻秸发酵液的显著性差异物种(Biomarker)增加;稻秸发酵液与瘤胃液亲缘关系较近,而添加乙醇显著改变了细菌区系;短链脂肪酸比例在稻秸发酵液细菌群落多样性中具有重要作用。研究表明,体外频繁传代和添加乙醇可以提高稻秸发酵液的乙酸、戊酸和己酸产量,乙醇改变了稻秸发酵液的细菌群落结构。     

葛根芩连汤在模拟胃肠液中对微生物生长的影响

目的探究葛根芩连汤在模拟胃肠液中对微生物生长的影响。方法在模拟胃肠液中分别加入消化液10%量的金黄色葡萄球菌、大肠埃希菌、沙门菌、产气杆菌、黑曲霉、米曲霉、青霉和汉逊德巴利酵母,然后加入生药浓度为1 g/mL的葛根芩连汤水煎液,37℃恒温培养。分别在0、1、2和4 h吸取1 mL培养液做稀释平板计数。结果体外模拟肠液中,葛根芩连汤对金黄色葡萄球菌、大肠埃希菌、沙门菌、产气杆菌、黑曲霉、米曲霉和青霉有较强的抑制作用,而对汉逊德巴利酵母的生长则为先促进后抑制,并且在2 h时对黑曲霉、米曲霉、青霉和汉逊德巴利酵母的抑制效果最显著;在模拟胃液中,葛根芩连汤对金黄色葡萄球菌、大肠埃希菌、沙门菌、黑曲霉、青霉和汉逊德巴利酵母均有一定的抑制作用,而对产气杆菌无明显影响。结论葛根芩连汤在模拟胃肠液中对金黄色葡萄球菌、大肠埃希菌、沙门菌、黑曲霉、米曲霉、青霉和汉逊德巴利酵母等均具有一定的抑制作用,在模拟肠液中对产气杆菌抑制作用明显,而在模拟胃液中对其抑制效果不明显。     

Effectiveness of host cell protein removal using depth filtration with a filter containing diatomaceous earth

The increased cell density and product titer in biomanufacturing have led to greater use of depth filtration as part of the initial clarification of cell culture fluid, either as a stand-alone unit operation or after centrifugation. Several recent studies have shown that depth filters can also reduce the concentration of smaller impurities like host cell proteins (HCP) and DNA, decreasing the burden on subsequent chromatographic operations. The objective of this study was to evaluate the HCP removal properties of the Pall PDH4 depth filter media, a model depth filter containing diatomaceous earth, cellulose fibers, and a binder. Experiments were performed with both cell culture fluid (CCF) and a series of model proteins with defined pI, molecular weight, and hydrophobicity chosen to match the range of typical HCP. The location of adsorbed (fluorescently labeled) proteins within the depth filters was determined using confocal scanning laser microscopy. Protein binding was greater for proteins that were positively charged and more hydrophobic, consistent with adsorption to the negatively charged diatomaceous earth. The lowest degree of binding was seen with proteins near their pI, which were poorly removed by this filter. These results provide new mechanistic insights into the factors governing the filter capacity and performance characteristics of depth filters containing diatomaceous earth that are widely used in the clarification of CCF.