泡桐愈伤组织再生植株的诱导与培养

首先以MS为基本培养基从18个不同浓度的NAA和BA组合中,找出了毛泡桐（Paulownia tomentosa）、南方泡桐（Pauiownia australis）、白花泡桐（Paulownia fortunei）、兰考泡桐（Paulownia elongata）和豫杂一号泡桐(Paulownia tomentosa×P.fortunei)叶片愈伤组织诱导芽分化最适培养基分别为MS+0.3NAA+12 BA、MS+0.3NAA+12 BA、MS+0.5NAA+12 BA、MS+0.5NAA+12 BA和MS+0.7NAA+12 BA;然后,筛选出了5种泡桐芽诱导根的最适培养基（分别为1/2MS+0.1NAA、1/2MS+0.1NAA、1/2MS、1/2MS+0.3NAA和1/2MS+0.5NAA）。这些结果为利用不同种泡桐的原生质体融合培育泡桐新品种奠定了一定的基础。     

N-acetylaspartate: a literature review of animal research on brain ischaemia

The present review examines and discusses the changes in N-acetylaspartate (NAA) concentration in the ischaemic brain from the existing literature of animal research. By summarizing the current knowledge on NAA metabolic pathways and reviewing the data obtained in animal models of global and focal ischaemia, the following conclusions emerge from this compilation: (i) both magnetic resonance spectroscopy (MRS) and the absolute HPLC method of NAA quantification give converging information; (ii) decreases in brain NAA concentration in acute stroke can be considered as an index of neuronal loss or dysfunction, although NAA redistribution by glial cells and NAA trapping in cell debris restrict its use as a quantitative neuronal marker; (iii) further studies on NAA metabolism in pathologic brain are required before using NAA measurement in the chronic stage of ischaemia for evaluating neuroprotective strategies.     

An anti-inflammatory role for N-acetyl aspartate in stimulated human astroglial cells

Although N-acetyl-L-aspartate (NAA) has been shown to be important to myelin synthesis and osmotic regulation, the biological rationale for the high levels of NAA found in the brain remains unknown. Here, a human astroglial cell line (STTG) was treated with NAA and stimulated with ionomycin, ionomycin/PMA, or IL-1beta. PGE(2) levels in ionomycin-stimulated STTG cells decreased by 76% and > 95% at NAA concentrations of 10 and 20mM, respectively. NAA also decreased the levels of COX-2 protein and activated NF-kappaB in IL-1beta-stimulated STTG cells but had little effect on unstimulated cells. Also, NAA significantly decreased intracellular calcium levels in ionomycin/PMA-stimulated cells. NAA had no effect on total COX-2 activity or COX-2 mRNA. Acetylation of IkappaBalpha kinase, an acetylation target of aspirin, was not observed when NAA was present. These results demonstrate that NAA appears to be important in the modulation of inflammation in the human STTG astroglial cell line. The results of these findings are discussed in relation to neuronal pathologies that exhibit abnormal NAA levels within the brain.     

Ovarian Cyst Fluid of Serous Ovarian Tumors Contains Large Quantities of the Brain Amino Acid N-acetylaspartate

Background

In humans, N-acetyl L-aspartate (NAA) has not been detected in other tissues than the brain. The physiological function of NAA is yet undefined. Recently, it has been suggested that NAA may function as a molecular water pump, responsible for the removal of large amounts of water from the human brain. Ovarian tumors typically present as large cystic masses with considerable fluid accumulation.

Methodology and Principal Findings

Using Gas Chromatography-Mass Spectrometry, we demonstrated that NAA was present in a high micromolar concentration in oCF of epithelial ovarian tumors (EOTs) of serous histology, sometimes in the same range as found in the extracellular space of the human brain. In contrast, oCF of EOTs with a mucinous, endometrioid and clear cell histological subtype contained a low micromolar concentration of NAA. Serous EOTs have a cellular differentiation pattern which resembles the lining of the fallopian tube and differs from the other histological subtypes. The NAA concentration in two samples of fluid accumulation in the fallopian tube (hydrosalpinx) was in the same ranges as NAA found in oCF of serous EOTs. The NAA concentration in oCF of patients with serous EOTs was mostly 10 to 50 fold higher than their normal serum NAA concentration, whereas in patients with other EOT subtypes, serum and cyst fluid NAA concentration was comparable.

Conclusions and Significance

The high concentration of NAA in cyst fluid of serous EOTs and low serum concentrations of NAA in these patients, suggest a local production of NAA in serous EOTs. Our findings provide the first identification of NAA concentrations high enough to suggest local production outside the human brain. Our findings contribute to the ongoing research understanding the physiological function of NAA in the human body.     

Investigation into the Role of N-Acetylaspartate in Cerebral Osmoregulation

Abstract: Marked abnormalities of the magnetic resonance intensity of N -acetylaspartate (NAA) have been reported in patients with various neurological disorders, but the neurochemical consequences of these alterations are difficult to assess because the function of NAA remains speculative. The purpose of this study was to examine whether NAA plays a role in protecting neurons against osmotic stress. Intracerebral microdialysis was used to expose a small region of the rat dorsolateral striatum to an increasingly hyposmotic environment and to measure resulting changes in NAA extracellular concentrations. NAA changes in the extracellular fluid (ECF) were compared with those of the amino acids, in particular, taurine, known to be involved in brain osmoregulation. Stepped increases in cellular hydration produced by hyposmotic perfusion media induced a marked increase in ECF NAA, reflecting a redistribution of NAA from intra-to extracellular space. Parallel experiments showed that, of all the extracellular amino acids measured, only taurine markedly increased with hyposmolar perfusion medium, indicating that the ECF NAA increase associated with hyposmotic stress was a specific response and not passive leakage out of the cells. As NAA is predominantly neuronal, it may contribute to the protection of neurons against swelling (i.e., regulatory volume decrease). In conditions with impaired blood-brain barrier and cytotoxic oedema, efflux of intracellular NAA subsequent to sustained cellular swelling might lead to a reduction in total brain NAA detectable by magnetic resonance spectroscopy. Alternatively, redistribution of NAA from intra-to extracellular space implies changes in its chemical environment that may alter its magnetic resonance visibility.     

N-ACETYL-l-ASPARTIC ACID CONTENT OF HUMAN NEURAL TUMOURS AND BOVINE PERIPHERAL NERVOUS TISSUES

Abstract— Abstract-Tumors of the human nervous system were utilized to investigate the cellular distribution of N-acetyl-L-aspartic acid (NAA). Astroglial tumours contained about 0.144 μmol/g. Oligodendrogliomas and medulloblastomas contained somewhat larger amounts. However, the level of NAA in all gliomas studied was less than that of normal human white matter. NAA was undetectable in meningiomas and acoustic neurinomas. If these results may be taken as representative of normal tissue, they imply a predominantly neuronal localization for NAA.
Substantial amounts of NAA were found in peripheral nervous tissues and retina. Neurons seem to vary widely in NAA content.     

Regulation of N-acetylaspartate and N-acetylaspartylglutamate biosynthesis by protein kinase activators

The neuronal dipeptide N-acetylaspartylglutamate (NAAG) is thought to be synthesized enzymatically from N-acetylaspartate (NAA) and glutamate. We used radiolabeled precursors to examine NAA and NAAG biosynthesis in SH-SY5Y human neuroblastoma cells stimulated with activators of protein kinase A (dbcAMP; N6,2'-O-dibutyryl cAMP) and protein kinase C (PMA; phorbol-12-myristate-13-acetate). Differentiation over the course of several days with dbcAMP resulted in increased endogenous NAA levels and NAAG synthesis from l-[(3)H]glutamine, whereas PMA-induced differentiation reduced both. Exogenously applied NAA caused dose dependent increases in intracellular NAA levels, and NAAG biosynthesis from l-[(3)H]glutamine, suggesting precursor-product and mass-action relationships between NAA and NAAG. Incorporation of l-[(3)H]aspartate into NAA and NAAG occurred sequentially, appearing in NAA by 1 h, but not in NAAG until between 6 and 24 h. Synthesis of NAAG from l-[(3)H]aspartate was increased by dbcAMP and decreased by PMA at 24 h. The effects of PMA on l-[(3)H]aspartate incorporation into NAA were temporally biphasic. Using short incubation times (1 and 6 h), PMA increased l-[(3)H]aspartate incorporation into NAA, but with longer incubation (24 h), incorporation was significantly reduced. These results suggest that, while the neuronal production of NAA and NAAG are biochemically related, significant differences exist in the regulatory mechanisms controlling their biosynthesis.     

Dynamic or inert metabolism? Turnover of N‐acetyl aspartate and glutathione from d‐[1‐13C]glucose in the rat brain in vivo

The rate of (13)C-label incorporation into both aspartyl (NAA C3) and acetyl (NAA C6) groups of N-acetyl aspartate (NAA) was simultaneously measured in the rat brain in vivo for up to 19 h of [1-(13)C]glucose infusion (n = 8). Label incorporation was detected in NAA C6 approximately 1.5 h earlier than in NAA C3 because of the delayed labeling of the precursor of NAA C3, aspartate, compared to that of NAA C6, glucose. The time courses of NAA were fitted using a mathematical model assuming synthesis of NAA in one kinetic compartment with the respective precursor pools of aspartate and acetyl coenzyme A (acetyl-CoA). The turnover rates of NAA C6 and C3 were 0.7 +/- 0.1 and 0.6 +/- 0.1 micromol/(g h) with the time constants 14 +/- 2 and 13 +/- 2 h, respectively, with an estimated pool size of 8 micromol/g. The results suggest that complete label turnover of NAA from glucose occurs in approximately 70 h. Several hours after starting the glucose infusion, label incorporation into glutathione (GSH) was also detected. The turnover rate of GSH was 0.06 +/- 0.02 micromol/(g h) with a time constant of 13 +/- 2 h. The estimated pool size of GSH was 0.8 micromol/g, comparable to the cortical glutathione concentration. We conclude that NAA and GSH are completely turned over and that the metabolism is extremely slow (< 0.05% of the glucose metabolic rate).     

Evidence supporting a role for N-acetyl-L-aspartate as a molecular water pump in myelinated neurons in the central nervous system. An analytical review.

Molecular water pumps (MWPs) are characterized as biochemical systems existing at a compartmental boundary of living cells that can actively pump water against its gradient. A role for the observed intercompartmental transport of N-acetyl-L-aspartate (NAA), between neurons and oligodendrocytes in the CNS, as an efflux MWP for the removal of neuronal metabolic water has been proposed. In this review, accumulating evidence in support of such a role for NAA is presented, and the dynamics of the NAA cycle in myelinated neurons are considered. Based on the results of recent investigations, it is calculated that 1 mol of NAA is synthesized for every 40 mol of glucose (Glc) equivalent oxidized in the brain, and each mol of NAA may transport 121 mol of metabolic water out of neurons. In addition, turnover of total brain NAA is very rapid and appears to be only 16.7 h. Thus, the most important characteristic of NAA in the brain may not be its static level, but a dynamic aspect related to its rapid turnover. The relationship of NAA as a potential MWP to Canavan disease (CD), a genetic spongiform leukodystrophy in which the catabolic portion of the NAA cycle is deficient, and in a newly recognized brain disorder, hypoacetylaspartia, where the anabolic portion of the NAA cycle appears to be deficient, are discussed.     

Direct determination of the N-acetyl-L-aspartate synthesis rate in the human brain by (13)C MRS and [1-(13)C]glucose infusion

A non-invasive (13)C magnetic resonance spectroscopy (MRS) technique is described for the determination of the N-acetyl-L-aspartate (NAA) synthesis rate, V(NAA), in the human brain in vivo. In controls, the mean V(NAA) was 9.2 +/- 3.9 nmol/min/g. In Canavan disease, where [NAA] is increased (p < 0.001) and [aspartate] is deceased (p < 0.001), V(NAA) was significantly reduced to 3.6 +/- 0.1 nmol/min/g (p < 0.001). These rates are in close agreement with the activity of the biosynthetic enzyme measured in vitro in animals, and with the rate of urinary excretion of NAA in human subjects with Canavan disease. The present result is consistent with the regulation of NAA synthesis by the activity of a single enzyme, L-aspartate-N-acetyltransferase, in vivo, and with its control in Canavan disease by limited substrate supply and/or product inhibition. The (13)C MRS technique provides the means for further determination of abnormal rates of neuronal NAA synthesis among neurological disorders in which low cerebral [NAA] has been identified.     

PFN2 and NAA80 cooperate to efficiently acetylate the N-terminus of actin

The actin cytoskeleton is of profound importance to cell shape, division, and intracellular force generation. Profilins bind to globular (G-)actin and regulate actin filament formation. Although profilins are well-established actin regulators, the distinct roles of the dominant profilin, profilin 1 (PFN1), versus the less abundant profilin 2 (PFN2) remain enigmatic. In this study, we use interaction proteomics to discover that PFN2 is an interaction partner of the actin N-terminal acetyltransferase NAA80, and further confirm this by analytical ultracentrifugation. Enzyme assays with NAA80 and different profilins demonstrate that PFN2 binding specifically increases the intrinsic catalytic activity of NAA80. NAA80 binds PFN2 through a proline-rich loop, deletion of which abrogates PFN2 binding. Small-angle X-ray scattering shows that NAA80, actin, and PFN2 form a ternary complex and that NAA80 has partly disordered regions in the N-terminus and the proline-rich loop, the latter of which is partly ordered upon PFN2 binding. Furthermore, binding of PFN2 to NAA80 via the proline-rich loop promotes binding between the globular domains of actin and NAA80, and thus acetylation of actin. However, the majority of cellular NAA80 is stably bound to PFN2 and not to actin, and we propose that this complex acetylates G-actin before it is incorporated into filaments. In conclusion, we reveal a functionally specific role of PFN2 as a stable interactor and regulator of the actin N-terminal acetyltransferase NAA80, and establish the modus operandi for NAA80-mediated actin N-terminal acetylation, a modification with a major impact on cytoskeletal dynamics.     

Natural autoantibodies of neonatal rats detectable by a hybridoma technic

Neonatal rat hybridomas were tested for natural autoantibodies (NAA) production, using different screening procedures. NAA were discovered in 35% of immunoglobulins producing hybridomas. Radioimmunoassay (RIA) on brain and liver homogenates and immunocytochemistry on brain sections are the procedures of choice revealing the major part of the identified NAA. On the contrary, only a small portion of NAA could be detected with indirect immunofluorescence on fixed fibroblasts and with RIA on individual autoantigens. All the NAA revealed proved to be of the IgM type and almost all of them possessed neither organ nor species-specificity. In spite of that, most of the NAA reacted with definite cell populations of nervous tissue such as glia, neurons, ependyma or brain vessel cells. The studied panel of NAA from neonatal rats has common features with similar panels from newborn and old mice, though some species-specific characteristics do exist.     

Some Physical-kinetic Considerations in Penetration of Naphthaleneacetic Acid through Isolated Pear Leaf Cuticle

The penetration of naphthaleneacetic acid (NAA) through enzymatically isolated pear leaf cuticle (Pyrus communis L. cv. Bartlett) is reported herein. Penetration of NAA increased with increasing lime and attained a steady state in approximately 20 minutes. The quantity of NAA penetrating was directly related to the concentration of the donor solution. NAA that penetrated the cuticle was shown to he unaltered. The Penetration of NAA from inside to outside is similar to that from outside to inside. Isolated stomatous lower cuticle permitted approximately 10-foId greater penetration of NAA than the astomatous upper cuticle. The penetration of NAA through isolated pear leaf cuticle is highly temperature dependent, exhibiting a temperature coefficient (Q₁₀) of about 5.6 between 15° and 25 C. The low quantities of chemicals penetrating through the isolated cuticle reported herein and elsewhere are considered to he a characteristic of the technique and not an absolute limitation of the cuticle. Cuticular penetration could account for physiological quantities of NAA entering the plant.     

Effect of N-acetylaspartic acid on the diffusion coefficient of water: a proton magnetic resonance phantom method for measurement of osmolyte-obligated water

N-acetyl-L-aspartic acid (NAA) is an amino acid present in the vertebrate brain that is synthesized and stored primarily in neurons, although it cannot be hydrolyzed in these cells. Nonetheless, neuronal NAA is dynamic and turns over more than once each day by cycling, via extracellular fluids (ECF), between neurons and catabolic compartments in oligodendrocytes. One important role of the NAA intercompartmental cycle appears to be osmoregulatory, and in this role it may be the primary mechanism for the removal of metabolic water, against a water gradient, from myelinated neurons. However, the number of water molecules that might be cotransported to ECF per NAA molecule released is as yet unclear. In this investigation, using a proton nuclear magnetic resonance method and diffusion measurements at two magnetic field strengths on water and NAA phantoms in vitro, the effect of NAA on the diffusion coefficient of water has been measured, and a ratio (K) of obligated water molecules per molecule of NAA has been determined. For NAA measured at 100mM and 3 Tesla K=24 and at 7 Tesla K=14. Based on these results, apparent K(NAA) varies inversely with field strength, and with a computed field strength factor of 2.55mmol water/unit Tesla, K(NAA) in the absence of any applied magnetic field strength would be 32.     

Extracellular N-Acetylaspartate in the Rat Brain: In Vivo Determination of Basal Levels and Changes Evoked by High K+

Abstract: The purpose of this study was to determine the extracellular concentrations of N -acetylaspartate (NAA) in the rat cerebral cortex, striatum, and hippocampus of halo-thane-anaesthetised rats by intracerebral microdialysis, and to examine the effects of high K⁺-induced local depolarisation, which provokes synchronous neurotransmitter release, cell swelling, and acid-base changes. Basal levels of NAA in the extracellular fluid (EOF) were determined by the zero net flux method. Tissue levels of NAA in the cortex, striatum, and hippocampus were 8.4, 5.7, and 7.2 mmol/kg, respectively. The corresponding extracellular concentrations of NAA were much lower (35.1, 83.7, and 23.0 tiM). High tissue/ECF concentration ratios may suggest little release or leakage of NAA under basal conditions, and potent reuptake mechanisms for NAA in the cellular membrane of CNS cells. There was no change in ECF NAA during K⁺-induced local depolarising stimuli produced in the striatum, but NAA levels consistently increased after the K⁺ stimuli, irrespective of whether or not Ca²⁺ was present in the perfusion medium. These data confirm that NAA is not a neurotransmitter and suggest strongly that NAA is not directly involved in the release and reuptake or metabolism of neuroactive compounds. The increase of NAA in the ECF immediately after K⁺ stimulation may reflect an involvement in brain osmoregulation and/or acid-base homeostasis.     

Specific Expression of N-Acetylaspartate in Neurons, Oligodendrocyte-Type-2 Astrocyte Progenitors, and Immature Oligodendrocytes In Vitro

To test the specificity of N-acetylaspartate (NAA) as a neuronal marker for proton nuclear magnetic resonance (1H NMR) spectroscopy, purified and characterized cultured cells were analyzed for their NAA content using both 1H NMR and HPLC. Cell types studied included cerebellar granule neurons, type-1 astrocytes, meningeal cells, oligodendrocyte-type-2 astrocyte (O-2A) progenitor cells, and oligodendrocytes. A high concentration of NAA was found in extracts of cerebellar granule neurons (approximately 12 nmol/mg of protein), whereas NAA remained undetectable in purified type-1 astrocytes, meningeal cells, and mature oligodendrocytes. However, twice the neuronal level of NAA was found in O-2A progenitors grown in vitro. In addition significant levels of NAA were also detected in cultures of immature oligodendrocytes. Our data partly support previous suggestions that NAA may be a useful neuronal marker for 1H NMR spectroscopic examination of the adult brain. However, they also raise the further possibility that alterations of NAA associated with some specific brain disorders, particularly disorders seen in newborn and young children, may reflect abnormalities in the development of oligodendroglia or their precursors.     

Studies on the Role of N-Acetylaspartic Acid in Mammalian Brain

N-Acetylaspartic acid (NAA) occurs at relatively high concentrations exclusively in the mammalian and avian brain and undergoes rapid rise in level soon after birth (Tallan, 1957). The amount of NAA in brains of mentally abnormal human beings and of young human beings was measured. The route by which NAA is synthesized was shown to involve a direct acetylation of aspartic acid. The degradative activity of the brain toward NAA is slight. Some experiments indicate that NAA in the brain is a physiologically and metabolically active compound.     

Mast Cells Contain Large Quantities of Secretagogue-Sensitive N-Acetylaspartate

Abstract: Mast cells play a central role in both immediate allergic reactions and inflammation. A functional nerve-mast cell interaction has been proposed, given the morphological association between mast cells and neuropeptide-containing peripheral nerves. We now show that purified rat peritoneal mast cells contain large quantities of N -acetylaspartate (NAA; 747.50 nmol/mg of protein). Mast cell levels of NAA were rapidly reduced, by 64.0 and 86.4%, following treatment with compound 48/80 and mastoparan, respectively. These secretagogues strongly decreased mast cell histamine content over the same time period, suggesting also that NAA is stored in secretory granules. The data are the first to show that NAA is present in an immune effector cell type. Because NAA may be involved in myelin synthesis and glutamyl peptide metabolism, NAA released from mast cells following nervous or other stimuli could participate in neuroimmune interactions. Mast cells in multiple sclerosis plaques may contribute to the reported elevations in brain NAA in this disease.     

磁共振波谱分析在颅脑胶质瘤分级中的应用研究

目的 分析脑胶质瘤的氢质子磁共振波谱（proton magnetic resonance spectroscopy,1H-MRS）表现及其临床意义;探讨脑胶质瘤的1H-MRS特点与其病理级别相关性.方法 搜集经临床手术、病理证实的脑胶质瘤病例49例,按照WHO诊断标准分成两组：低级别脑胶质瘤组、高级别脑胶质瘤组.所有患者在术前行1H-MRs检查,均在MR非增强成像的基础上获得.使用Philips Achieva 1.5T超导磁共振扫描仪,单体素或多体素扫描,点分辨法,检测不同区域代谢物变化.结果 脑胶质瘤的1H-MRS表现：肌酸（Cr）轻度下降,N-乙酰天门冬氨酸（NAA）显著下降,胆碱（Cho）显著增高.低、高级别脑胶质瘤的肿瘤组织与对侧止常脑组织的NAA、Cho、NAA/Cr、NAA/Cho值存在显著性差异（P〈0.05）;低级别和高级别脑胶质瘤的肿瘤组织的NAA/Cr、NAA/Cho值存在显著性差异（P〈0.05）.脑胶质瘤的NAA/Cho、Cho/Cr、NAA/Cr值与病理级别相关,其中NAA/Cho和NAA/Cr值反映肿瘤级别较稳定;NAA/Cr、NAA/Cho值呈负相关关系,Cho/Cr值呈正相关关系.结论 ：1H-MRS结合MKI能提高脑胶质瘤术前诊断的准确性.1H-MRS能对胶质瘤进行分级,反映胶质瘤代谢特性以及肿瘤生长潜能.