The unprocessed C-terminal dipeptide of recombinant beta-nerve growth factor determines three stable forms with distinct biological activities.

The processing of polypeptide neurotrophins in the nervous system is poorly understood. In this paper, we provide information on the effects of C-terminal processing of nerve growth factor. Three forms of recombinant mouse beta-nerve growth factor (rNGF) were produced and isolated from insect cells infected with a recombinant baculovirus. The three purified forms of rNGF exhibited distinct biological activities and differed in their abilities to compete with high affinity binding of mouse beta-nerve growth factor (mNGF). However, they were chemically and structurally indistinguishable from each other. All three forms of rNGF differed from mature mNGF from mouse submaxillary gland in that the C-terminal Arg-Gly dipeptide had not been proteolytically removed. Removal of the C-terminal dipeptide by gamma-NGF peptidase treatment converted the three forms into a single form identical with mature mNGF. The above results demonstrate that a single polypeptide of rNGF, due to the presence of a C-terminal dipeptide, exhibits three stable dimeric protein conformations with distinct biological activities. The apparent lack of gamma-NGF peptidase in the nervous system raises the possibility that the biologically significant form of NGF may differ from mature mNGF; such a difference may be of physiological relevance.     

Spatial d/h heterogeneity of leaf water

The mean δD value of petiole water of Pterocarpus indicus Willd (δD = −9.0 ± 2.5‰, n = 3) was not significantly different from the mean value of stem water (−8.3 ± 2.8‰, n = 3). δD values of main vein water ranged from −11.1 to + 12.0‰ (n = 14) and increased along the main vein from petiole to the tip of leaves. Mesophyll water was highly enriched in deuterium (mean δD = +32.0 ± 2.0‰, n = 19) when compared with stem, petiole, and vein water. δD values of mesophyll water for different areas of the lamina, however, were not homogenous and could differ by as much as 20‰.     

In vitro expression and site-specific mutagenesis of the cloned human lipoprotein lipase gene. Potential N-linked glycosylation site asparagine 43 is important for both enzyme activity and secretion

Detailed structure-function information about human lipoprotein lipase (LPL) is unavailable because it is difficult to purify large amounts of the enzyme for study. To circumvent this problem, we constructed an in vitro LPL expression vector. Human LPL cDNA was cloned and inserted into the expression vector p91023(B). After transfection of COS M-6 cells with the human LPL cDNA construct, LPL enzyme activity was detected in cell extracts and culture medium. Purified human apolipoprotein C-II caused a 5-fold stimulation of the recombinant human LPL expressed in vitro. Using site-specific mutagenesis, Ala residues were substituted for Asn residues at two potential N-linked glycosylation sites (positions 43 and 359) and at a third unrelated Asn (position 257) in the LPL cDNA. RNA blot analysis demonstrated the presence of a single mRNA species in COS cells transfected with wild-type and mutant LPL expression vectors. Intracellular and secreted LPL activity was absent in the construct containing an Ala for Asn mutation at position 43, whereas the same substitutions at positions 257 and 359 did not appreciably affect activity. LPL activity was also absent in another construct containing a Gln for Asn mutation at position 43. Quantitation of LPL protein mass concomitant with measurement of enzyme activity showed that substitution of Ala or Gln for Asn at position 43 resulted in the production of an enzymatically inactive protein which accumulated intracellularly but was not secreted into the culture medium. Our report represents an initial documentation of the expression of cloned human LPL in vitro and of the importance of Asn-43 for both enzyme activity and secretion.     

Urinary excretion of a novel hexasaccharide and glycopeptide analogue in fucosidosis.

Repeated Biogel P6 chromatography of the urine from a patient with fucosidosis yielded several fractions containing fucosyloligosaccharides and glycopeptides. Two of these were shown by ¹H nuclear magnetic resonance (¹H-n.m.r.) spectroscopy and permethylation analysis to have the following structures respectively: (I) αfuc (1→3) [βgal (1→4)] βglcNAc (1→2) αman ($\text{1→}\text{3}\text{6}$) βman (1→4) glcNAc and (II) αfuc (1→3) [βgal (1→4)] βglcNAc (1→2) αman ($\text{1→}\text{3}\text{6}$) βman (1→4) βglcNAc (1→4) [αfuc ($\text{1→}\text{3}\text{6}$)] βglcNAc-Asn.     

六种海桑属红树植物根际真菌特征及其影响因子

真菌多样性是植物根际生态系统的重要构成与植物健康稳定的重要指标。海桑属是红树林的先锋物种,采用真菌ITS1区高通量测序方法,分析了六种海桑属红树根际真菌的组成和多样性,结合土壤理化性质探讨影响不同植物根际真菌群落组成差异的因素。结果显示,根际真菌隶属于7门、96科、155属,子囊菌门作为优势菌门在海桑属不同红树中相对丰度无显著差异,都超过27%,但次优势的担子菌门丰度含量有差异;属水平上,优势菌属的丰度含量不同,曲霉属在卵叶海桑的丰度最高(29.57%),在海南海桑最低(3.47%)。六种红树植物根际存在特有的代表类群,如无瓣海桑的马拉色菌(9.31%)和毛腐菌属(10.05%),海南海桑中的Talaromyces(19.61%)和Acremonium(13.58%)。比较多样性指数Simpson和Shannon,发现拟海桑是六种植物中丰度最高的,卵叶海桑最低。RDA分析发现子囊菌门与全磷含量呈显著负相关,担子菌门与速效钾呈明显正相关。六种海桑属红树植物根际核心物种分析表明,优势真菌类群曲霉属和一些低丰度的真菌类群,通过降解有机质参与碳循环,对根际土壤生态系统的稳定起重要作用。六种海桑...     

三峡库区消落带植被NPP估算——基于机器学习优化CASA模型

三峡库区蓄水后,其生态效应受到广泛关注。消落带植被固碳量作为衡量库区生态系统健康状态的重要指标,对库区碳循环与生态净化具有重要意义。针对消落带不同高程植被接受光照的时间有所差异,且受河流水位变化影响,传统的CASA模型在计算消落带植被固碳量时,存在对植物的光能利用率计算不够精确等问题。以三峡库区香溪河陡坡消落带为研究区域,提出了一种耦合RBFNN模型（Radial Basis Function Neural Network）与CASA模型（Carnegie-Ames-Stanford approach）的新方法（RBF-CASA）。基于RBFNN建立环境影响因子模型,借助高程数据及植被指数等特征计算适合消落带区域的环境影响因子。结合CASA模型中温度和水分胁迫因子,提高植被在像元尺度上的净初级生产力（Net Primary Productivity,NPP）的估算精度,并对反演结果进行验证。模型验证结果显示：RBF-CASA模型估算值与观测值的决定系数（Coefficient of determination, R²）为0.730（P<0.01, n=32）。对比原始CASA模型,平均绝对误差（Mean absolute error, MAE）降低10.991,均方根误差（Root mean square error, RMSE）降低了23.861,相对均方根误差（Relative root mean square error, RRMSE）降低5.10%,平均绝对百分误差（Mean absolute percentage error, MAPE）降低1.12%。使用提出的RBF-CASA模型在库区水位落干期（7-8月份）进行固碳量估算,结果表明：NPP月均值在66.234-134.144g C/m²之间,NPP随着高程的增加呈现起伏变化,其总量在150-155m之间达到峰值,均值在170m以上区域最高。在2021年9月植被NPP均值为35.883g C/m²,2022年9月植被NPP均值为25.964g C/m²,由于降雨量减少、长江水位下降,在2021-2022年间植被恢复情况较差。研究结果可为库区碳循环、生态净化及生态修复等决策提供科学依据。