Digestibility, nutrient balance and urinary purine derivative excretion in dry yak cows fed oat hay at different levels of intake

A feeding trial A as conducted at the farm of Qinghai Academy of Animal and Veterinary Science, Xining, China during 1996 - 1997 with three dry yak cows (initial body weight 163 - 197 kg, age 5 - 6 years) by using 3 x 3 Latin Square Design to determine the effect of levels of feed intake on digestion, nitrogen balance and purine derivative excretion in urine of yak cows. The animals were fed oat hay (nitrogen 13.5 g/kg dry matter (DM), metabolisable energy 8.3 MJ/kg DM), i.e., 0.3, 0.6 and 0.9 of voluntary intake (VI). Each intake treatment lasted for 17 days and the samples (feeds, faeces and urine) were collected during last 7 days of each period. The results indicate that digestibility of dietary DM, OM, NDF and ash declined when intake levels increased from 0.3 to 0.9 VI [DM, from 66.1% to 59.1% (P < 0.05); OM, from 68.1% to 59.9% (P < 0.05); NDF, from 62.1% to 54.3% (P < 0.05); and ash, from 33.9% to 11.8% (P < 0.05)]. Around 0.10 g N/kg W-0.75 was deficient daily in yak cows at 0.3 VI, and positive N balances were observed at 0.6 and 0.9 VI. Intake levels significantly (P < 0.05) affected total PD excretion in yak urine. The proportion of allantoin increased (P < 0.05) and uric acid decreased (P < 0.05) as intake level of feed increased. (C) 2004 Elsevier B.V. All rights reserved.

中国中部全新世气候变化的泥炭同位素记录--以湖北神农架大九湖为例

泥炭作为一种地质档案，其发育形成过程中包含了丰富的古气候和古环境的变化信息。由于泥炭沉积中植物残体的碳、氧等稳定同位素组成与气候变化之间存在着密切关系，其稳定同位素时间序列已作为气候变化的代用指标逐渐用于古气候的恢复和重建，尤其是在以高时间分辨率重建全新世以来的气候环境变化方面已取得一些重要进展。 本论文通过对湖北省神农架地区大九湖泥炭中植物残体的氧、碳同位素的研究，同时以温度和湿度这两种重要的气候参数代用指标，恢复和重建全新世以来中国中部地区的气候状况。通过本论文的工作并与其它相关成果综合对比研究，对从末次冰消期晚期和全新世以来中国中部地区气候状况有如下的认识： 1. 末次冰消期晚期至全新世阶段，大九湖泥炭纤维素δ18O敏感地记录了发生在约14.67～12.89 cal. kaBP的BA暖期以及在此期间所发生的两次冷事件：Oder Dryas冷事件和Intra-Allerød冷事件；并指示随后的Allerød暖期明显较Bølling暖期偏冷；在新仙女木时期（YD），神农架地区温度较低，事件中后期温度开始回升；在11.0～9.0 cal. kaBP的早全新世时期，大九湖泥炭δ18O在波动中升高，指示了整体上的温暖气候，期间存在一系列的气候变冷事件。这些冷事件基本都是叠加在一种长期的、相对较暖的变化趋势上，有别于末次冰消期的突然变冷事件；在9.0～5.0 cal. kaBP的中全新世阶段，神农架地区总体上处于相对温暖的适宜阶段，但波动较为频繁，期间记录到了著名的“8.2ka”冷事件；约5.0 cal. kaBP开始，大九湖泥炭δ18O值持续升高，指示以一次快速的升温过程开始了晚全新世阶段并保持在一个相对较高的温度水平；在整体温暖的气候背景下也明显记录有“4.2ka”、“小冰期”等全球性冷事件；大九湖泥炭纤维素δ18O所记录的温度变化趋势与中国大陆地区其他一些气候代用指标所记录的温度变化趋势有很好的相似性，说明中国大陆地区温度变化的趋势基本一致。 2. 大九湖泥炭δ18O温度变化序列所记录到的历次突然气候变冷事件，与格陵兰冰芯和北大西洋冷事件的发生具有很好的一致性，这说明对应于北半球高纬度区域的气候变化，在中低纬度的中国地区同样存在响应和变化，并且这种响应贯穿于整个全新世阶段。 3. 大九湖泥炭纤维素δ13C敏感地记录了末次冰消期以来东亚季风的演变历史。东亚季风在整个末次冰消期至晚全新世阶段始终处于一种强弱交替的波动态势。在末次冰消期至全新世早期，其整体上表现为一种由弱转强的态势；在全新世早期和中期阶段，东亚季风活动强度整体较高，表现为整个时间序列中的高活动时段，而其中也出现了数次明显的减弱迹象；全新世晚期阶段，东亚季风的活动强度较中期和早期为弱，整体上表现出低幅的波动但没有恢复到前两个时期的活动强度。在中世纪温暖期阶段的中后期，东亚季风的活动强度开始在波动中逐渐升高。 4. 该泥炭纤维素代用指标所记录的一些东亚季风突变（增强）事件与同一时期所发生的印度季风的突变（减弱）事件具有很好的对应关系；进一步证实这两个季风系统之间所存在的反相变化关系；同时，这些突变事件与北大西洋IRD冷事件在发生时间段也具有较好的对应关系，即每当北大西洋地区气候变冷时，东亚季风都表现出增强的迹象。 5. 综合分析中国泥炭同位素古气候记录和其它研究结果，对气候驱动机制有如下认识：当太阳活动发生百年至千年时间尺度的减弱时，由大气臭氧层变化所触发的影响向大气层下部延伸，导致北半球气候变冷；同时，减弱的太阳活动可能在赤道太平洋触发类厄尔尼诺状态出现，并引发印度洋夏季风减弱和东亚夏季风增强；早全新世北半球冰盖周围大冰湖冰岸坍塌，湖中巨量体积淡水突然流入北大西洋，导致海洋温盐环流传送带运转放慢甚致停顿，通过跨赤道的温盐环流向北输送的热量大大减少，引起北半球气候变冷；由于海洋温盐环流的变化改变了全球热量的分配，暖水在地球南部积累，导致西太平洋暖池区域海表面水与深层水温度之间反差的增加，从而有利于类厄尔尼诺状态出现，同样将可能引起印度洋夏季风减弱和东亚夏季风增强。

单种属泥炭纤维素碳稳定同位素组成与全新世青藏高原东部西南季风变化

泥炭的形成和积累主要受控于气候，其次是地质、地貌和水文等因素。作为一种重要和理想的气候信息载体，泥炭己经与冰芯、海洋及湖泊沉积、黄土堆积、洞穴碳酸盐沉积、树轮和珊瑚礁等其它类型的自然地质档案一样，逐渐为国内外致力于古气候变化研究的学者所接受，并将其重点应用于全新世以来气候变化信息的提取。本研究在已有的泥炭混合植物残体纤维素碳稳定同位素记录青藏高原东部地区气候信息的基础上，首次提取泥炭中单一种属植物残体一木里苔草（Calexmulieensis）纤维素，测定其碳稳定同位素，并以其时间序列作为一种新的西南季风代用指标，揭示该区域全新世以来气候变化。研究表明：1．木里苔草的碳稳定同位素时间序列是西南季风强度变化的敏感代用指标。约11800～11加0 cal aBP期间，木里苔草纤维素的613C记录处于最大值，表明该时段西南季风活动鼻弱，气候干冷，对应于普遍发生并存在的新仙女木事件；它清晰地指示了该区全新世的下限年龄为约11200 cal aBP（14c年龄约9900aB玲从约11200 oal aBP起该区迅速进入湿暖的全新世阶段，季风活动迅速增强；在约10800～5500 cal aBP期间，季风总体保持在强盛状态，但其间有4次突然减弱，气候变干冷；约从5500 cala即起季风活动在波动中逐渐减弱，其中有4次减弱最为明显。2．木里苔草纤维素碳同位素所记录的犯000年以来的所有9次西南季风活动的突然减弱与同时期北大西洋发生的冰力｝漂移碎屑沉积物事件（IRD事件）一一对应。这种密切的相关关系表明，西南季风强度的波动可能是对全球气候变化特别是对海洋热盐环流引起的地球南北方气撇动变化的响应。3．木里苔草纤维素碳稳定同位素所指示的西南季风有952，乓57，475，312，211和77年等一系列周期。其中557年和77年两个周期分别与北大西洋深层海水环流的550年的周期和76年的气候周期非常吻合。这表明西南季风与北大西洋气候间有很好的相关关系。由于557年和77年的季风周期又都分别类似于53。年和80年的太阳活动周期，这表明太阳活动可能对西南季风和北大西洋气候间的联系有着影响。4．红原泥炭木里苔草纤维素和泥炭混合纤维素别3c时间序列在千年至万年时间尺度上的变化趋势很相近。两种代用记录都一致反映了冰后期的气候变化，即均指示西南季风强度变化可分为三个明显的大阶段。这说明它们对过去12000年的西南季风变化具有相同的响应。对于目前经过较详细研究的那些全球大范围发生的突然气候变化事件，木里苔草纤维素δ~(13)C代用记录与泥炭混合纤维素δ~(13)C代用记录一样，都有相同的响应，但是，相对于泥炭中单一的木里苔草敏感地记录到历次北大西洋I劝事件而言，泥炭混合纤维素δ~(13)C记录对其中的几次突然气候变化事件似乎没有表现出明显的响应。这个结果表明对数百年尺度的突然气候变化，木里苔草纤维素比混合纤维素的酬3c指标似乎要更敏感一点。因此，在能获得足够数量的单种植物·残体的情况下，单种植物残体纤维素的δ~(13)C指标有很好的使用价值。

Removal of endotoxin from human serum albumin solutions by hydrophobic and cationic charged membrane

A novel matrix of macropore cellulose membrane was prepared by chemical graft, and immobilized the cationic charged groups as affinity ligands. The prepared membrane Fan be used for the removal of endotoxin from human serum albumin (HSA) solutions. With a cartridge of 20 sheets affinity membrane of 47 mm diameter, the endotoxin level in HSA solution can be reduced ro 0.027 eu/mL. Recovery of HSA was over 95%.

Modeling of protein adsorption in membrane affinity chromatography

For the design of affinity membranes, protein adsorption in membrane affinity chromatography (MAC) was studied by frontal analysis. According to fast mass transfer, small thickness of affinity membranes and high affinity between the protein and the ligand, an ideal adsorption (IA) model was proposed for MAC and was used together with equilibrium-dispersive (E-D) model to describe the adsorption of bovine serum albumin (BSA) onto cellulose diacetate/polyethyleneimine (CA/PEI) blend membranes with and without Cu2+ chelating. E-D model was found to better describe the initial region of experimental breakthrough curves. The influence of axial dispersion was revealed and it showed the importance of design of the module to homogenously distribute feed solution. IA model was found to be better for the whole experimental breakthrough curve. According to it, the capacity of affinity membranes and the specificity of the interaction are of equal importance for the design of affinity membranes. An optimum feed concentration was also found in the operation of MAC. The discrepancy between experimental optimum feed concentrations and predicted ones from IA model may be due to the ignorance of some experimental effects such as axial dispersion.

Morphology transcription process from CMC micelles to inorganogel and its effect on the properties of alumina particle

In this study, a carboxymethyl cellulose (CMC)-mediated sol-gel process was developed to synthesize the alumina hydoxide whiskers. During the process, inexpensive inorganic salts were used as precursors and supercritical drying method was used to extract the water in hydrogel. The influences of CMC on the gel formation and the particle morphology were investigated. The results show that the formation of CMC-aluminium hydroxide organic-inorganic hybridgels led to a morphology transcription process from CMC micelles to aluminium hydroxide gel, as a result, the precursor with whiskerious morphology was obtained.

Rapid protein identification using monolithic enzymatic microreactor and LC-ESI-MS/MS

A monolithic enzymatic microreactor was prepared in a fused-silica capillary by in situ polymerization of acrylamide, glycidyl methacrylate (GMA) and ethylene dimethacrylate (EDMA) in the presence of a binary porogenic mixture of dodecanol and cyclohexanol, followed by ammonia solution treatment, glutaraldehyde activation and trypsin modification. The choice of acrylamide as co-monomer was found useful to improve the efficiency of trypsin modification, thus, to increase the enzyme activity. The optimized microreactor offered very low back pressure, enabling the fast digestion of proteins flowing through the reactor. The performance of the monolithic microreactor was demonstrated with the digestion of cytochrome c at high flow rate. The digests were then characterized by CE and HPLC-MS/MS with the sequence coverage of 57.7%. The digestion efficiency was found over 230 times as high as that of the conventional method. in addition, for the first time, protein digestion carried out in a mixture of water and ACN was compared with the conventional aqueous reaction using MS/MS detection, and the former solution was found more compatible and more efficient for protein digestion.

Tratamento de dejetos no âmbito do programa do ABC.

2015

Caratterizzazione proteomica di fluidi e tessuti in diverse condizioni fisio-patologiche

This thesis has been focused on the proteomic characterization of human saliva from donors of different ages, starting from birth up to adult age, and pediatric brain tumor tissues. The first study has been performed in order to compare the acid-insoluble fraction of saliva from preterm with at-term newborns and adults and establish if differences exist. In the second study medulloblastoma and pilocytic astrocytoma pediatric brain tumor extracts have been compared. In both studies 2- DE analysis was coupled with high resolution tandem mass spectrometry (MS/MS). The proteomic characterization of the acid-insoluble fractions of saliva from preterm newborns allowed to integrate data previously obtained on the acid-soluble fraction by HPLC-electrospray ionization (ESI)-mass spectrometry (MS), and to evidence several differences between preterm newborns, at-term newborns and adults. Spots differentially expressed between the three groups, according to image analysis of the gels, were submitted to in-gel tryptic digestion and the peptide mixture analyzed by high performance HPLC-ESI-MS/MS for their characterization. By this strategy, we identified three over-expressed proteins in atterm newborns with respect to preterm newborns and adults (BPI fold-containing family A member 1, two proteoforms of annexin A1, and keratin type 1 cytoskeletal 13), and several over-expressed proteins in adults (fatty acid-binding protein, S100A6, S100A7, two proteoforms of S100A9, several proteoforms of prolactin-inducible protein, Ig kappa chain, two proteoforms of cystatin SN, one proteoform of cystatin S and several proteoforms of α-amylase 1). Moreover, for the first time, it was possible to assign by MS/MS four spots of human saliva 2-DE, already detected by other authors, to different proteoforms of S100A9. The strategy applied used a sequential staining protocol to the 2-DE gels, first with Pro-Q Diamond, that allows specific detection of phosphoproteins, and successively with total protein SYPRO Ruby stain. In the second study, proteomic analysis of two pediatric brain tumor tissues pointed out differences between medulloblastoma, the prevalent malignant tumor in childhood, and pilocytic astrocytoma, the most common, that only rarely shows a malignant progression. Due to the limited availability of bioptic tissue, the study was performed on pooled tumor tissues, and was focused on acid-insoluble fraction to integrate the characterization performed by a group of colleagues in Rome on the acid-soluble fraction by high performance HPLC-ESI-MS/MS. The results indicated that the two tumors exhibit different proteomic profiles and evidenced interesting differential expression of several proteins. Among them, peroxiredoxin- 1, peptidyl-prolyl cis–trans isomerase A, heterogeneous nuclear ribonucleoproteins A2/B1, mitochondrial isoform of malate dehydrogenase, nucleoside diphosphate kinase A, glutathione S-transferase P and fructose bisphosphate aldolase A resulted significantly over-expressed in medulloblastoma while glial fibrillary acidic protein, serotransferrin, α crystallin B chain, ferritin light chain, annexin A5, fatty acid-binding protein (brain), sorcin and apolipoprotein A-I resulted significantly over-expressed in pilocytic astrocytoma. In conclusion, the work done allowed to evidence the usefulness of using an integrated bottom-up/top-down approach, based on 2-DE-MS analysis and high performance MS in order to obtain a complete characterization of the proteome under investigation, revealing and identifying, not only peptides and small proteins, but also proteins with higher MW, that often it is not possible to identify by using exclusively a top-down ESI-MS approach.

Dynamic effects of cofactors and DNA on the oligomeric state of human mitochondrial DNA helicase

We examined the effects of cofactors and DNA on the stability, oligomeric state and conformation of the human mitochondrial DNA helicase. We demonstrate that low salt conditions result in protein aggregation that may cause dissociation of oligomeric structure. The low salt sensitivity of the mitochondrial DNA helicase is mitigated by the presence of magnesium, nucleotide, and increased temperature. Electron microscopic and glutaraldehyde cross-linking analyses provide the first evidence of a heptameric oligomer and its interconversion from a hexameric form. Limited proteolysis by trypsin shows that binding of nucleoside triphosphate produces a conformational change that is distinct from the conformation observed in the presence of nucleoside diphosphate. We find that single-stranded DNA binding occurs in the absence of cofactors and renders the mitochondrial DNA helicase more susceptible to proteolytic digestion. Our studies indicate that the human mitochondrial DNA helicase shares basic properties with the SF4 replicative helicases, but also identify common features with helicases outside the superfamily, including dynamic conformations similar to other AAA⁺ ATPases.

Aqueous solutions of HEC and hmHEC: effects of molecular mass versus hydrophobic associations on hydrodynamic and thermodynamic parameters.

Aqueous solutions of amphiphilic polymers usually comprise of inter- and intramolecular associations of hydrophobic groups often leading to a formation of a rheologically significant reversible network at low concentrations that can be identified using techniques such as static light scattering and rheometry. However, in most studies published till date comparing water soluble polymers with their respective amphiphilic derivatives, it has been very difficult to distinguish between the effects of molecular mass versus hydrophobic associations on hydrodynamic (intrinsic viscosity [g]) and thermodynamic parameters (second virial coefficient A2), owing to the differences between their degrees of polymerization. This study focuses on the dilute and semi-dilute solutions of hydroxyethyl cellulose (HEC) and its amphiphilic derivatives (hmHEC) of the same molecular mass, along with other samples having a different molecular mass using capillary viscometry, rheometry and static light scattering. The weight average molecular masses (MW) and their distributions for the nonassociative HEC were determined using size exclusion chromatography. Various empirical approaches developed by past authors to determine [g] from dilute solution viscometry data have been discussed. hmHEC with a sufficiently high degree of hydrophobic modification was found to be forming a rheologically significant network in dilute solutions at very low concentrations as opposed to the hmHEC with a much lower degree of hydrophobic modification which also enveloped the hydrophobic groups inside the supramolecular cluster as shown by their [g] and A2. The ratio A2MW/[g], which takes into account hydrodynamic as well as thermodynamic parameters, was observed to be less for associative polymers compared to that of the non-associative polymers.

X-ray simulations for imaging applications in the agricultural and food industry

R. Zwiggelaar, C.R. Bull, and M.J. Mooney, 'X-ray simulations for imaging applications in the agricultural and food industry', Journal of Agricultural Engineering Research 63(2), 161-170 (1996)

Molecular analysis of the anaerobic rumen fungus Orpinomyces - insights into an AT-rich genome

Matthew J. Nicholson, Michael K. Theodorou and Jayne L. Brookman. (2005). Molecular analysis of the anaerobic rumen fungus Orpinomyces - insights into an AT-rich genome. Microbiology, 151 (1), 121-133. Sponsorship: BBSRC RAE2008

Towards the identification of the common features of bacterial biofilm development

UPNa. Instituto de Agrobiotecnología. Laboratorio de Biofilms Microbianos