BACKGROUND & AIMS: :pH 5.9 is optimal for tobacco pollen tube growth in suspension culture. Little pH changes of sugar-mineral medium result from the release of surface-linked compounds from pollen grains. Germination and pollen tube growth bring about a progressive medium acidification resulting in total growth inhibition. An increase of the buffering capacity of the culture medium enhances pollen tube growth. When the pH was kept near the optimum by 25 mM MES-KOH buffer, pollen tubes grew for 4 days and in the presence of casein hydrolysate they reached a length of up to about 4 cm. The growth related acidification of the medium is independent of the presence of mineral cations and is not due to the hydration of respiratory CO(2). It is suggested that it is brought about by proton secretion from pollen tubes.

背景与目标： : pH 5.9对于悬浮培养中的烟草花粉管生长是最佳的。糖矿物培养基的ph值变化很小，这是由于花粉粒中表面连接的化合物的释放所致。发芽和花粉管生长会导致培养基酸化，从而导致总生长抑制。培养基缓冲能力的增加可增强花粉管的生长。当通过25mmmes-KOH缓冲液将pH保持在最佳值附近时，花粉管生长4天，并且在酪蛋白水解产物存在下，它们达到高达约4厘米的长度。与生长相关的培养基酸化与矿物阳离子的存在无关，而不是由于呼吸CO(2) 的水合作用。建议它是由花粉管的质子分泌引起的。

BACKGROUND & AIMS: OBJECTIVES:To determine whether vascular endothelial growth factor A (VEGF) and its receptors are expressed during bladder development in mice when capillaries are forming, and whether exogenous VEGF might enhance the growth of endothelia and other types of bladder cells, using an embryonic organ-culture model. MATERIALS AND METHODS:Whole bladders from wild-type mice, at embryonic day (E) 14, were grown in serum-free organ culture in an air/5% CO2 atmosphere; some cultures were supplemented with VEGF and/or with VEGF receptor 1/Fc chimera (VEGFR1/Fc), which blocks VEGF bioactivity. Organs were harvested after 6 days and the expression of VEGF and related molecules assessed using immunohistochemistry. RESULTS:VEGF, VEGFR1 and VEGFR2 positive cells were immunodetected in E14 and E18 bladders. Exogenous VEGF increased whole-organ growth, as assessed by explant areas, total cell numbers, DNA and protein content; proliferation was enhanced, and apoptosis decreased, in urothelium and surrounding tissues. VEGF also increased the proportions of cells expressing endothelial (CD31) and smooth muscle (alpha smooth muscle actin) markers. VEGFR1/Fc blocked the growth-enhancing effects of exogenous VEGF. CONCLUSIONS:In organ culture, exogenous VEGF not only stimulated embryonic bladder endothelial cells but also strikingly enhanced the growth of the whole organ. Whether the effects of VEGF on diverse bladder cell populations are direct or indirect requires further investigation. The finding that VEGF protein is present in embryonic bladders in vivo raises the possibility that it has similar actions during normal development. The results also illuminate the pathobiology of certain bladder diseases in which VEGF levels have been shown to be increased.

背景与目标：

BACKGROUND & AIMS: :Culture-based studies of the microbial community within the gut of the medicinal leech have typically been focused on various Aeromonas species, which were believed to be the sole symbiont of the leech digestive tract. In this study, analysis of 16S rRNA gene clone libraries confirmed the presence of Aeromonas veronii and revealed a second symbiont, clone PW3, a novel member of the Rikenellaceae, within the crop, a large compartment where ingested blood is stored prior to digestion. The diversity of the bacterial community in the leech intestinum was determined, and additional symbionts were detected, including members of the alpha-, gamma-, and delta-Proteobacteria, Fusobacteria, Firmicutes, and Bacteroidetes. The relative abundances of the clones suggested that A. veronii and the novel clone, PW3, also dominate the intestinum community, while other clones, representing transient organisms, were typically present in low numbers. The identities of these transients varied greatly between individual leeches. Neither time after feeding nor feeding on defibrinated blood caused a change in identity of the dominant members of the microbial communities. Terminal restriction fragment length polymorphism analysis was used to verify that the results from the clone libraries were representative of a larger data set. The presence of a two-member bacterial community in the crop provides a unique opportunity to investigate both symbiont-symbiont and symbiont-host interactions in a natural model of digestive-tract associations.

背景与目标： : 对药用水蛭肠道内微生物群落的基于培养的研究通常集中在各种气单胞菌物种上，这些物种被认为是水蛭消化道的唯一共生体。在这项研究中，对16S rRNA基因克隆文库的分析证实了veronii气单胞菌的存在，并揭示了第二个共生体，即Rikenellaceae的新成员clone PW3，在作物内有一个大的隔间，在消化之前会储存摄入的血液。确定了水蛭肠道中细菌群落的多样性，并检测到了其他共生体，包括 α-，γ-和 δ-变形菌，梭菌，厚壁菌和拟杆菌的成员。克隆的相对丰度表明，A. veronii和新型克隆PW3也主导着肠群落，而代表瞬态生物的其他克隆通常以少量存在。这些瞬变的身份在各个水ches之间差异很大。喂食或喂食脱纤维的血液后的时间都不会导致微生物群落主要成员的身份发生变化。末端限制性片段长度多态性分析用于验证克隆库的结果代表较大的数据集。作物中两人细菌群落的存在为在消化道关联的自然模型中研究共生体-共生体和共生体-宿主相互作用提供了独特的机会。

BACKGROUND & AIMS: :Complete nucleotide sequence of the tissue culture-adapted ATCC-Wa strain of human rotavirus NSP4 was determined. Sequence analysis detected two alternate forms of the gene with a nucleotide difference at position 331 (A or G) in the coding sequence (NSP4-A or NSP4-G) leading to a change from neutral glutamine97 in NSP4-A to a positively charged arginine97 in NSP4-G originating from the same ATCC-Wa preparation. In addition to this, both forms of ATCC-Wa NSP4 revealed three mutations at nucleotide positions 88 (T to C), 142 (C to T) and 572 (G to A), when compared to the previously reported NSP4 sequence from virulent Wa strain. The former two mutations were in the coding sequence and resulted in a leucine16 to serine16 and a proline34 to leucine34 change, while the third mutation was in the 3' non-coding region of the gene. The two amino acid changes may contribute to the 'tissue culture-adaptation' of ATCC-Wa strain. The ATCC-Wa NSP4 sequence was found to differ from the previously reported NSP4 sequence of attenuated Wa strain by lacking a mutation at 133 (T to C), though the mutations at 88 and 142 were present in both strains. Furthermore, comparison of deduced amino acid sequence of NSP4 from human, bovine, porcine and simian rotavirus strains identified a seven-residue (positions 135-141) inter-species variable domain in its C-terminal region.

背景与目标： : 确定了人类轮状病毒NSP4的组织培养适应的atcc-wa株的完整核苷酸序列。序列分析检测到在编码序列 (NSP4-A或NSP4-G) 中位置331 (a或G) 处具有核苷酸差异的基因的两种替代形式，导致从NSP4-A的中性谷氨酰胺97变为源自相同atcc-wa制剂的NSP4-G中的带正电荷的精氨酸97。除此之外，当与先前报道的来自强毒Wa株的NSP4序列相比时，两种形式的atcc-wa NSP4在核苷酸位置88 (T至C) 、142 (C至T) 和572 (G至A) 处揭示了三个突变。前两个突变在编码序列中，导致亮氨酸16到丝氨酸16和脯氨酸34到亮氨酸34的变化，而第三个突变在基因的3' 非编码区。这两个氨基酸的变化可能有助于atcc-wa菌株的 “组织培养适应”。发现atcc-wa NSP4序列与先前报道的减毒Wa菌株的NSP4序列不同，因为在133 (T至C) 处没有突变，尽管在88和142处的突变存在于两个菌株中。此外，从人，牛，猪和猿猴轮状病毒株中推导的NSP4氨基酸序列的比较确定了其C末端区域中的七个残基 (位置135-141) 种间可变结构域。

BACKGROUND & AIMS: :The development of more productive strains of microorganisms and processes that increase enzyme levels can contribute to the economically efficient production of second generation ethanol. To this end, cellulases and xylanases were produced with the S1M29 mutant strain of Penicillium echinulatum, using different concentrations of cellulose (20, 40, and 60 g L(-1)) in batch and fed-batch processes. The highest activities of FPase (8.3 U mL(-1)), endoglucanases (37.3 U mL(-1)), and xylanases (177 U mL(-1)) were obtained in fed-batch cultivation with 40 g L(-1) of cellulose. The P. echinulatum enzymatic broth and the commercial enzyme Cellic CTec2 were tested for hydrolysis of pretreated sugar cane bagasse. Maximum concentrations of glucose and xylose were achieved after 72 h of hydrolysis. Glucose yields of 28.0% and 27.0% were obtained using the P. echinulatum enzymatic extract and Cellic CTec2, respectively.

背景与目标： : 开发更具生产力的微生物菌株和提高酶水平的工艺可以有助于经济高效地生产第二代乙醇。为此，在分批和补料分批过程中，使用不同浓度的纤维素 (20、40和60g L(-1))，用棘青霉的S1M29突变菌株生产纤维素酶和木聚糖酶。在用40g L(-1) 纤维素补料分批培养中获得最高的FPase (8.3 U mL(-1)) 、内切葡聚糖酶 (37.3 U mL(-1)) 和木聚糖酶 (177 U mL(-1)) 活性。测试了echinulatum酶促肉汤和商业酶Cellic CTec2对预处理的甘蔗渣的水解作用。水解72小时后达到最大浓度的葡萄糖和木糖。分别使用棘皮草酶提取物和Cellic CTec2获得28.0% 和27.0% 的葡萄糖产量。

BACKGROUND & AIMS: :Spheroids are three-dimensional cell aggregates expressing histotypic organisation in vitro comparable to tissue continuity in vivo. They can be prepared from normal tissue and from tumour fragments. In the experiments presented here, dermal human spheroids and brain tumour spheroids are prepared from the same patient. The dermal tissue originates from the border of the incision wound made to effect a stereotactic brain tumour biopsy. The tumour originates from a fragment of the collected stereotactic biopsy. The dermal fragment and the brain biopsy are explanted in vitro to form confluent monolayers. At confluency, the dermal cells are transferred into small Erlenmeyer flasks and rotated at 37 degrees C for 1-2 days and rotation mediated spheroids are formed. Small flaps of the tumour monolayer are placed on a semisolid non-adhesive substrate, reorganise and form agar overlay spheroids. After spheroid formation, a dermal spheroid is confronted with a brain tumour derived spheroid. The confronting pair, after adhering to each other, present an invasion model in vitro. The dermal spheroid functions as the autologous host for the brain tumour spheroid. Putative invasive cells present in the reaggregated brain spheroid will invade the dermal spheroid and destroy it. If no invasive cells are present in the tumour derived spheroid no morphologic changes will be seen in the dermal spheroid; 24 tested brain biopsy spheroids demonstrated a clear correlation between malignancy in situ and invasiveness in vitro. So it can be concluded that the autologous confrontation of brain tumour derived spheroids with dermal spheroids derived from the patient has a predictive value concerning malignant evolution and mimics the situation of the tumour in situ.

背景与目标： : 球体是三维细胞聚集体，在体外表达组织类型，与体内组织连续性相当。它们可以从正常组织和肿瘤碎片中制备。在这里介绍的实验中，皮肤人球体和脑肿瘤球体是由同一患者制备的。真皮组织起源于切口伤口的边界，以进行立体定向脑肿瘤活检。肿瘤起源于收集的立体定向活检的片段。在体外移植真皮碎片和脑活检以形成汇合的单层。汇合时，将真皮细胞转移到小锥形瓶中，并在37 ℃ 旋转1-2天，并形成旋转介导的球体。将肿瘤单层的小皮瓣放置在半固体非粘附基质上，重组并形成琼脂覆盖球体。球体形成后，真皮球体面对脑肿瘤衍生的球体。相互粘附后，面对的一对在体外呈现入侵模型。真皮球体充当脑肿瘤球体的自体宿主。在重新聚集的大脑球体中存在的假定的侵入性细胞将侵入真皮球体并破坏它。如果肿瘤衍生的球体中没有浸润性细胞，则真皮球体中不会出现形态学变化; 24个经过测试的脑活检球体表明原位恶性肿瘤与体外侵袭性之间存在明显的相关性。因此可以得出结论，脑肿瘤衍生的球体与患者衍生的真皮球体的自体对抗具有有关恶性演变的预测价值，并模仿了原位肿瘤的情况。

BACKGROUND & AIMS: Excitatory amino-acid currents in rodent central neurones are mediated by the activation of glutamate receptors. Ionotropic types of the receptors are divided into alpha-amino-3-hydroxy-5-methyl-4-isoxazolepropionate (AMPA), kainate and N-methyl-D-aspartate (NMDA) receptors, and the former two are collectively called non-NMDA receptors. The NMDA receptor is modulated by a number of endogenous neuromodulators including Mg2+, polyamines, glycine and protons in extracellular solutions. Although it has been generally thought that each of the neuromodulators acts on a distinct site in the NMDA receptor, recent studies have revealed that these actions may be not necessarily independent of each other. The NMDA receptor response is not only inhibited but also potentiated by Mg2+, and the latter action is due to an interaction of a Mg2+ site with either glycine- or proton-binding site. In the presence of polyamines, a tonic inhibition by protons of the NMDA receptor response is relieved, resulting in a potentiation of the response. Alternatively, it has been recently revealed that there are some subtypes of non-NMDA receptors which are negatively modulated by polyamines in either extra- or intra cellular solutions. The difference in polyamine sensitivity among non-NMDA receptors is attributed to a distinction in their constituted subunits. The inhibition of non-NMDA receptor by intracellular polyamines results in inward rectification of the current-voltage relation which is not seen for polyamine-insensitive ones. This polyamine action is not mimicked by intracellular Mg2+.

背景与目标： 啮齿动物中枢神经元中的兴奋性氨基酸电流由谷氨酸受体的激活介导。离子型受体分为alpha-amino-3-hydroxy-5-methyl-4-isoxazolepropionate (AMPA)，kainate和N-甲基-D-天冬氨酸 (NMDA) 受体，前两者统称为非NMDA受体。NMDA受体受到许多内源性神经调节剂的调节，包括细胞外溶液中的Mg2，多胺，甘氨酸和质子。尽管人们普遍认为每种神经调节剂都作用于NMDA受体的不同部位，但最近的研究表明，这些作用可能不一定彼此独立。NMDA受体反应不仅受到抑制，而且还受到Mg2的增强，后者的作用是由于Mg2位点与甘氨酸或质子结合位点的相互作用。在存在多胺的情况下，质子对NMDA受体反应的强音抑制被缓解，从而导致反应增强。或者，最近已经发现，在细胞外或细胞内溶液中，有一些非NMDA受体的亚型被多胺负调节。非NMDA受体之间多胺敏感性的差异归因于其组成亚基的差异。细胞内多胺对非NMDA受体的抑制导致电流-电压关系的内向整流，这对于多胺不敏感的人是看不到的。细胞内Mg2不会模仿这种多胺作用。

BACKGROUND & AIMS: :Staphylococcus aureus (S. aureus) causes persistent biofilm-related infections. Biofilm formation by S. aureus is affected by the culture conditions and is associated with certain genotypic characteristics. Here, we show that glucose and sodium chloride (NaCl) supplementation of culture media, a common practice in studies of biofilms in vitro, influences both biofilm formation by 40 S. aureus clinical isolates (methicillin-resistant and methicillin-sensitive S. aureus) and causes variations in biofilm quantification. Methicillin-resistant strains formed more robust biofilms than methicillin-sensitive strains in tryptic soy broth (TSB). However, glucose supplementation in TSB greatly promoted and stabilized biofilm formation of all strains, while additional NaCl was less efficient in this respect and resulted in significant variation in biofilm measurements. In addition, we observed that the ST239-SCCmec (Staphylococcal Cassette Chromosome mec) type III lineage formed strong biofilms in TSB supplemented with glucose and NaCl. Links between biofilm formation and accessory gene regulator (agr) status, as assessed by δ-toxin production, and with mannitol fermentation were not found. Our results show that TSB supplemented with 1.0% glucose supports robust biofilm production and reproducible quantification of S. aureus biofilm formation in vitro, whereas additional NaCl results in major variations in measurements of biofilm formation.

背景与目标： : 金黄色葡萄球菌 (S. aureus) 引起持续的生物膜相关感染。金黄色葡萄球菌的生物膜形成受培养条件的影响，并与某些基因型特征有关。在这里，我们显示了培养基的葡萄糖和氯化钠 (NaCl) 补充，这是体外生物膜研究的常见做法，影响了40株金黄色葡萄球菌临床分离株 (耐甲氧西林和甲氧西林敏感金黄色葡萄球菌) 的生物膜形成，并导致生物膜定量的变化。在胰蛋白酶大豆肉汤 (TSB) 中，耐甲氧西林的菌株比对甲氧西林敏感的菌株形成更坚固的生物膜。然而，TSB中的葡萄糖补充极大地促进和稳定了所有菌株的生物膜形成，而额外的NaCl在这方面的效率较低，并导致生物膜测量的显着变化。此外，我们观察到ST239-SCCmec (葡萄球菌盒染色体mec) III型谱系在补充了葡萄糖和NaCl的TSB中形成了强生物膜。未发现通过 δ 毒素产生评估的生物膜形成与辅助基因调节剂 (agr) 状态以及甘露醇发酵之间的联系。我们的结果表明，补充有1.0% 葡萄糖的TSB支持体外稳定的生物膜产生和金黄色葡萄球菌生物膜形成的可重复定量，而额外的NaCl导致生物膜形成的测量结果的主要变化。

BACKGROUND & AIMS: AIM:NetworkZ is a simulation-based multidisciplinary team-training programme designed to enhance patient safety by improving communication and teamwork in operating theatres (OTs). In partnership with the Accident Compensation Corporation, its implementation across New Zealand (NZ) began in 2017. Our aim was to explore the experiences of staff - including the challenges they faced - in implementing NetworkZ in NZ hospitals, so that we could improve the processes necessary for subsequent implementation. METHOD:We interviewed staff from five hospitals involved in the initial implementation of NetworkZ, using the Organising for Quality model as the framework for analysis. This model describes embedding successful quality improvement as a process of overcoming six universal challenges: structure, infrastructure, politics, culture, motivation and learning. RESULTS:Thirty-one people participated. Structural support within the hospital was considered essential to maintain staff enthusiasm, momentum and to embed the programme. The multidisciplinary, simulation-based approach to team training was deemed a fundamental infrastructure for learning, with participants especially valuing the realistic in situ simulations and educational support. Participants reported positive changes to the OT culture as a result of NetworkZ and this realisation motivated its implementation. In sites with good structural support, NetworkZ implementation proceeded quickly and participants reported rapid cultural change towards improved teamwork and communication in their OTs. CONCLUSION:Implementation challenges exist and strategies to overcome these are informing future implementation of NetworkZ. Embedding the programme as business as usual across a nation requires significant and sustained support at all levels. However, the potential gains in patient safety and workplace culture from widespread multidisciplinary team training are substantial. Trial registration number ACTRN12617000017325.

背景与目标：

BACKGROUND & AIMS: :Further refinement of culture media is needed to improve the quality of embryos generated in vitro. Previous results from our laboratory demonstrated that uptake of nutrients by the embryo is significantly less than what is supplied in traditional culture media. Our objective was to determine the impact of reduced nutrient concentrations in culture medium on mouse embryo development, metabolism, and quality as a possible platform for next generation medium formulation. Concentrations of carbohydrates, amino acids, and vitamins could be reduced by 50% with no detrimental effects, but blastocyst development was impaired at 25% of standard nutrient provision (reduced nutrient medium; RN). Addition of pyruvate and L-lactate (+PL) to RN at 50% of standard concentrations restored blastocyst development, hatching, and cell number. In addition, blastocysts produced in RN + PL contained more ICM cells and ATP than blastocysts cultured in our control (100% nutrient) medium; however, metabolic activity was altered. Similarly, embryos produced in the RN medium with elevated (50% control) concentrations of pyruvate and lactate in the first step medium and EAA and Glu in the second step medium were competent to implant and develop into fetuses at a similar rate as embryos produced in the control medium. This novel approach to culture medium formulation could help define the optimal nutrient requirements of embryos in culture and provide a means of shifting metabolic activity towards the utilization of specific metabolic pathways that may be beneficial for embryo viability.

背景与目标： : 需要进一步细化培养基，以提高体外产生的胚胎的质量。我们实验室的先前结果表明，胚胎对营养的吸收明显少于传统培养基中提供的营养。我们的目标是确定培养基中营养浓度降低对小鼠胚胎发育，代谢和质量的影响，作为下一代培养基配方的可能平台。50% 可以降低碳水化合物，氨基酸和维生素的浓度，而没有有害作用，但是在标准营养素供应 (营养培养基减少; RN) 的25% 下，囊胚发育受损。以50% 标准浓度向RN添加丙酮酸和L-乳酸 (PL) 可恢复胚泡发育，孵化和细胞数量。此外，在rn   +  PL中产生的囊胚比在我们的对照 (100% 营养) 培养基中培养的囊胚含有更多的ICM细胞和ATP; 然而，代谢活性发生了改变。类似地，在RN培养基中产生的胚胎具有升高 (50% 对照) 浓度的丙酮酸和乳酸在第一步培养基中以及在第二步培养基中产生的EAA和Glu的胚胎能够以与在对照培养基中产生的胚胎相似的速率植入和发育成胎儿。这种新颖的培养基配方方法可以帮助确定培养物中胚胎的最佳营养需求，并提供一种将代谢活性转移到利用可能有益于胚胎生存的特定代谢途径的手段。

BACKGROUND & AIMS: :Fish sauce production relies on a natural fermentation process requiring 12-18 months for process completion. Virgibacillus sp. SK37 has been shown to be a potential strain for fish sauce acceleration. However, hydrolytic activity of proteinases bound at cell surface of this strain has not been well elucidated. Addition of 0.2 % CaCl(2) (w/w) in conjunction with starter cultures of Virgibacillus sp. SK 37 increased protein hydrolysis as measured by α-amino group content throughout fermentation (P < 0.05). Cell-bound proteinases from Virgibacillus sp. SK 37 were extracted into a free form by incubating the washed cells in Ca(2+)-free buffer at 37 °C for 2 h. Cell-bound proteinases revealed molecular mass of 19, 20, 22, 32, 34, and 44 kDa based on a synthetic peptide zymogram. The proteinases showed subtilisin-like serine characteristics with the highest activity at 50 °C and pH 8 and 11. Activity of the extracted proteinases increased ~4 times at ≥100 mM CaCl(2). In addition, CaCl(2) enhanced thermal stability of the extracted proteinases. Enzymes showed proteolytic activity in either the absence or presence of 10 and 25 % NaCl toward fish muscle, soy protein isolate, and casein substrates. Cell-bound proteinases were likely to play an important role in protein hydrolysis during fish sauce fermentation.

背景与目标： : 鱼露生产依赖于自然发酵过程，需要12-18个月才能完成过程。Virgibacillus sp。SK37已被证明是鱼露加速的潜在菌株。然而，尚未很好地阐明该菌株细胞表面结合的蛋白酶的水解活性。添加0.2% CaCl(2) (w/w) 与Virgibacillus sp. SK 37的发酵剂培养物一起增加了蛋白质水解，如在整个发酵过程中通过 α-氨基含量测量 (P <0.05)。通过将洗涤过的细胞在不含Ca(2) 的缓冲液中于37 °C孵育2小时，将来自Virgibacillus sp. SK 37的细胞结合蛋白酶提取为游离形式。根据合成肽酶谱，细胞结合蛋白酶的分子量为19、20、22、32、34和44 kDa。蛋白酶表现出枯草杆菌蛋白酶样丝氨酸特征，在50 °C和pH 8和11时活性最高。提取的蛋白酶的活性在 ≥ 100 mM CaCl(2) 时增加了约4倍。此外，CaCl(2) 增强了提取的蛋白酶的热稳定性。在不存在或存在10和25% NaCl的情况下，酶对鱼肌肉，大豆分离蛋白和酪蛋白底物表现出蛋白水解活性。在鱼露发酵过程中，细胞结合的蛋白酶可能在蛋白质水解中起重要作用。

BACKGROUND & AIMS: :This study endeavored to investigate the diversity of microbes present during the shaping, ripening and drying of Daqu, a fermentation starter culture and substrata complex of Maotai alcoholic spirit. A nested PCR-denaturing gradient gel electrophoresis technique was utilized with different combinations of primers. The results showed the presence of bacteria, yeasts and molds. The microflora, which originate from wheat, were readily detectable during every stage of the fermentation process. However, the microbial structure had clear differences in the shaping, ripening and drying processes. In the shaping stage, there was a high level of diversity of the LAB (lactic acid bacteria) and fungi in the shaped samples. In the ripening stage, however, a reduction of diversity of fungi with a high level of diversity of the Bacilli was observed in the ripened samples. In the drying stage, the diversity of Bacilli and fungi, especially acid-producing bacteria, reduced dramatically. Interestingly, uncultured Lactococcus sp., Microbacterium testaceum, Cochliobolus sp., and Thermoascus crustaceus were the first to be detected in the fermentation starters used in liquor production. This study revealed the microbial diversity and distributions during the shaping, ripening and drying of Daqu-making, facilitating evaluation of the hygienic conditions and aiding in the design of specific starter and/or adjunct cultures.

背景与目标： : 本研究致力于研究茅台酒精发酵发酵剂和基质复合物大曲的成型，成熟和干燥过程中存在的微生物多样性。将巢式PCR变性梯度凝胶电泳技术与不同的引物组合一起使用。结果表明存在细菌，酵母和霉菌。在发酵过程的每个阶段都很容易检测到源自小麦的微生物区系。然而，微生物结构在成型，成熟和干燥过程中存在明显差异。在成型阶段，成型样品中实验室 (乳酸菌) 和真菌的多样性很高。然而，在成熟阶段，在成熟的样品中观察到真菌多样性的减少以及细菌的高水平多样性。在干燥阶段，细菌和真菌，尤其是产酸细菌的多样性急剧减少。有趣的是，未培养的乳球菌属，微细菌testaceum，Cochliobolus sp。和甲壳纲Thermoascus是在白酒生产中使用的发酵发酵剂中首次检测到的。这项研究揭示了大曲成型，成熟和干燥过程中的微生物多样性和分布，有助于评估卫生条件，并有助于设计特定的发酵剂和/或辅助培养物。

BACKGROUND & AIMS: :The figure of the English bloodhound is often portrayed both positively and negatively as an efficient man-hunter. This article traces the cultural, social and forensic functions of the first attempts to use bloodhounds for police investigation, and argues that the analysis of these developments, which took place at the turn of the twentieth century, further our understanding of the diverse practices and cultures of fin de siècle forensics. Arguing that their dogs could trail tracks of human scent, English pedigree bloodhound breeders promoted and imagined novel ways of detecting and thinking forensically, with which they made claims to social authority in matters of crime and detection. Yet, English bloodhounds were unstable carriers of forensic meaning making their use for tracking criminals deeply problematic: for example, the name of the breed itself invoked a long-line of social and cultural associations. In showing this, we can see how the practices of canine forensics had their roots in a complex history, involving genteel leisure, changing cultural understandings of scent, and shifting dog-keeping mores.

背景与目标： : 英国猎犬的形象经常被正面和负面地描绘成一个高效的人类猎人。本文追溯了首次尝试使用猎犬进行警察调查的文化，社会和法医功能，并认为对这些发展的分析 (发生在20世纪之交) 进一步加深了我们对各种实践和文化的理解。fin desi è cle法医。英国血统的猎犬饲养者认为他们的狗可以追踪人类的气味，因此提倡并想象了新颖的法医侦查和思考方式，并以此来宣称他们在犯罪和侦查方面具有社会权威。然而，英国猎犬是法医意义上不稳定的载体，这使得它们用于追踪罪犯的行为严重成问题: 例如，该犬种本身的名称引起了一系列社会和文化联系。在展示这一点时，我们可以看到犬类法医的做法如何植根于复杂的历史，包括绅士休闲，改变对气味的文化理解以及改变养犬方式。

BACKGROUND & AIMS: :A Gram-stain-negative, non-motile, rod-shaped, red-pigmented, facultatively anaerobic bacterium, designated SS2-9T, was isolated from sediment collected from a sea cucumber culture pond located in Rongcheng, Shandong province, China. Cells of strain SS2-9T were approximately 0.3-0.5 µm in width and 1.5-6.0 µm in length. The strain was able to grow at 10-37 °C, at pH 6.5-8.5 and in the presence of 0.5-6.0 % (w/v) NaCl. It grew optimally at 28 °C and in the presence of 2.0 % (w/v) NaCl. The DNA G+C content was 34.5 mol% and the sole respiratory quinone was menaquinone 6 (MK-6). The predominant cellular fatty acids were C15 : 0, iso-C15 : 1 G, iso-C15 : 0 and iso-C17 : 0 3-OH. The major polar lipids were phosphatidylethanolamine, an unidentified phospholipid, two unidentified aminolipids and four unidentified lipids. Phylogenetic analyses based on 16S rRNA gene sequences revealed that strain SS2-9T was phylogenetically related to members of the genus Aquimarina and was closely related to Aquimarina amphilecti 92VT (97.29 % similarity). On the basis of phenotypic, chemotaxonomic and phylogenetic data, strain SS2-9T was considered to represent a novel species of the genus Aquimarina, for which the name Aquimarina rubra sp. nov. is proposed. The type strain is SS2-9T (=KCTC 52274T=MCCC 1H00142T).

背景与目标： : 从中国山东省荣成市海参养殖池的沉积物中分离出革兰氏染色阴性，非活动，杆状，红色色素，兼性厌氧细菌，命名为SS2-9T。菌株SS2-9T的细胞宽度约为0.3-0.5  µ m，长度约为1.5-6.0  µ m。该菌株能够在10-37 ℃ 、pH 6.5-8.5和0.5-6.0% (w/v) NaCl存在下生长。它在28  ℃ 和2.0   % (w/v) NaCl存在下最佳生长。DNA G + C含量为34.5  mol %，唯一的呼吸醌为menaquinone 6 (MK-6)。主要的细胞脂肪酸是c15 :   0，iso-C15  :  1G，iso-C15  :  0和iso-C17  : 0 3-OH。主要的极性脂质是磷脂酰乙醇胺，一种未鉴定的磷脂，两种未鉴定的氨基脂和四种未鉴定的脂质。基于16S rRNA基因序列的系统发育分析表明，SS2-9T菌株与Aquimarina属成员的系统发育相关，与Aquimarina amphilecti92vt (97.29   % 相似性) 密切相关。根据表型，化学分类学和系统发育数据，菌株SS2-9T被认为代表了Aquimarina属的一种新物种，其名称为Aquimarina rubra sp。11月。是提出的。类型应变是SS2-9T的 (= KCTC 52274T = mcc1h00142t)。

BACKGROUND & AIMS: :Serum of animal origin and in particular fetal bovine serum is the most commonly utilized cell culture medium additive for in vitro cell growth and differentiation. However, several major concerns have been raised by the scientific community regarding the use of animal sera for human cell-based culture applications. Among the possible alternatives to the animal serum, platelet-derived compounds have been proposed since more than 10 years. Nevertheless, the high degree of variability between the different platelet preparations, and the lack of standardized manufacturing and quality control procedures, made difficult to reach a consensus on the applicability of this novel cell culture medium supplement. In this study, we describe the preparation of a standardized platelet-rich plasma (PRP) derivative obtained starting from human-certified buffy coat samples with a defined platelet concentration and following protocols including also freeze-drying, gamma irradiation and biological activity testing prior the product release as cell culture medium additive. Biological activity testing of the different preparations was done by determining the capability of the different PRP preparations to sustain human bone marrow mesenchymal stem cell (MSC) clone formation and proliferation. Taking advantage of a developed MSC in vitro clonogenicity test, we also determined biological activity and stability of the freeze-dried gamma-sterilized PRP preparations after their storage for different times and at different temperatures. The PRP effects on cell proliferation were determined both on primary cell cultures established from different tissues and on a cell line. Results were compared with those obtained in "traditional" parallel control cultures performed in the presence of bovine serum [10% fetal calf serum (FCS)]. Compared to FCS, the PRP addition to the culture medium increased the MSC colony number and average size. In primary cell cultures and in cell line cultures, the PRP promoted cell proliferation also in conditions where the FCS had not a proliferation stimulating effect due to either the nature of the cells and the tissue of origin (such as human articular chondrocytes from elderly patients) or to the critical low density cell seeding (such as for HeLa cells). In summary, the standardized PRP formulation would provide an "off-the-shelf" product to be used for the selection and expansion of several cell types also in critical cell culture conditions.

背景与目标： : 动物来源的血清，特别是胎牛血清是最常用的细胞培养基添加剂，用于体外细胞生长和分化。然而，科学界对将动物血清用于基于人类细胞的培养应用提出了一些主要问题。在动物血清的可能替代品中，已有10多年的历史提出了血小板衍生化合物。然而，不同血小板制剂之间的高度差异，以及缺乏标准化的制造和质量控制程序，使得很难就这种新型细胞培养基补充剂的适用性达成共识。在这项研究中，我们描述了一种标准化的富血小板血浆 (PRP) 衍生物的制备，该衍生物是从具有确定的血小板浓度的人类认证的血沉棕黄涂层样品开始获得的，并且以下方案还包括冷冻干燥，γ 辐照和生物活性测试。产品作为细胞培养基添加剂释放。通过确定不同PRP制剂维持人骨髓间充质干细胞 (MSC) 克隆形成和增殖的能力，对不同制剂进行了生物活性测试。利用开发的MSC体外克隆性测试，我们还确定了冷冻干燥的 γ-灭菌PRP制剂在不同时间和不同温度下储存后的生物活性和稳定性。PRP对细胞增殖的影响是在从不同组织建立的原代细胞培养物和细胞系上确定的。将结果与在存在牛血清 [10% 胎牛血清 (FCS)] 的情况下进行的 “传统” 平行对照培养物中获得的结果进行比较。与FCS相比，向培养基中添加PRP增加了MSC菌落数和平均大小。在原代细胞培养和细胞系培养中，在FCS由于细胞的性质和起源组织 (例如老年患者的人关节软骨细胞) 或临界低密度细胞接种而没有增殖刺激作用的条件下，PRP也促进了细胞增殖 (例如HeLa细胞)。总之，标准化的PRP制剂将提供一种 “现成的” 产品，用于在临界细胞培养条件下选择和扩增几种细胞类型。