Enzymatic Synthesis of N-formyl-L-aspartyl-L-phenylalanine Methyl Ester (Aspartame Precursor) Utilizing an Extractive Reaction in Aqueous/Organic Biphasic Medium

N-Formyl-L-aspartyl-L-phenylalanine methyl ester (N-formyl aspartame, F-AspPheOMe) was synthesized enzymatically utilizing an extractive reaction in an aqueous/organic biphasic system. The N-formyl aspartame yield in a pure aqueous monophasic system was, in general, ca. 3% , however, it was over 80 % in a water/1-butanol biphasic system using a simultaneously extractive operation of an enzymatic reaction in an aqueous phase and a product separation from an aqueous to an organic phases.     

基于临床肿瘤标本的胃癌转移模型建立

目的建立基于临床肿瘤标本的胃癌转移模型,为胃癌的转移研究提供个体化动物模型。方法将胃癌新鲜的手术标本移植到裸鼠皮下,建立胃癌患者异种移植(patient-derived xenograft,PDX)模型。进一步通过手术将皮下瘤组织原位移植到裸鼠胃部肌层,连续观察裸鼠的体征状态,通过近红外荧光活体成像技术检测肿瘤转移的发生。解剖荷瘤小鼠,将肺部转移灶进一步移植裸鼠皮下获得实体瘤。HE染色观察原发瘤与转移瘤的结构特征,(short tandem repeat) STR分析原发瘤和转移瘤的遗传特性。PCR-Array分析转移瘤和原发瘤中转移相关基因的表达。结果成功建立胃癌PDX模型,移植瘤组织结构与患者保持基本一致;通过胃部原位移植发现编号C19751的小鼠发生肺和肝的转移。其中肺转移灶皮下移植后获得了实体瘤,STR分析显示原发瘤保持了与肺转移瘤一致的遗传特征。PCR-Array结果显示,与原发瘤相比,转移瘤中CXCL12,IGF1和MMP2基因表达均显著上调。结论利用临床肿瘤标本成功建立胃癌转移模型,为胃癌转移研究提供了良好的个体化模型。     

Array CGH characterization of an unbalanced X-autosome translocation associated with Xq27.2–qter deletion, 11q24.3–qter duplication and Xq22.3–q27.1 duplication in a girl with primary amenorrhea and mental retardation

We present array comparative genomic hybridization (aCGH) characterization of an unbalanced X-autosome translocation with an Xq interstitial segmental duplication in a 16-year-old girl with primary ovarian failure, mental retardation, attention deficit disorder, learning difficulty and facial dysmorphism. aCGH analysis revealed an Xq27.2–q28 deletion, an 11q24.3–q25 duplication, and an inverted duplication of Xq22.3–q27.1. The karyotype was 46,X,der(X)t(X;11)(q27.2;q24.3) dup(X)(q27.1q22.3). We discuss the genotype–phenotype correlation in this case. Our case provides evidence for an association of primary amenorrhea and mental retardation with concomitant unbalanced X-autosome translocation and X chromosome rearrangement.     

Suckling Behaviour in Natural and Artificially Fed Neonate Calves1

Das Trinken von Holstein- und Hereford-Kälbern an der Mutter und an Trinkapparaten wird beschrieben und verglichen. Die Kälber suchen nach der Geburt, sobald sie stehen können, nach der Milchquelle; sie saugen an der Maschine in gleicher Haltung wie an der Mutter. Kommt keine Milch, so lutschen sie dennoch am Sauger oder an Ohr, Skrotum, Euter oder an der Penis-scheide anderer Kälber. Dieses Ersatzlutschen verringert die Trockenfutteraufnahme erheblich. Am Trinkapparat saugen Holsteinkälber 16–24 Min./24 Std., Hereford-Kälber 22–42 Min./24 Std. (letztere an der Mutter 37–57 Min./24 Std.). Der Rasseunterschied hängt mit dem Körpergewicht und dieses mit der Trockenfutteraufnahme und der Wachstumsrate zusammen. Alle Kälber ergriffen die ihnen jeweils nächste Zitze zuerst, ohne eine bestimmte zu bevorzugen. An der Mutter saugten die Kälber 4-6mal pro Tag, am Trinkapparat mit verdünnter Milch bis zu 55mal; ein Paar Hereford-Zwillinge saugten an der Mutter 11 mal pro Tag, wohl weil deren Milch für zwei Junge etwas knapp war.     

Beobachtungen an sozialbezogenen Verhaltensweisen von Sammelbienen

Mit Hilfe neuer Registriermethoden wurden der Saugvorgang und das Duftmarkieren bei Sammelbienen in Abhängigkeit vom Zuckerwasserangebot an der Futterquelle untersucht. Wenn an einer Futterquelle das Angebot ergiebig ist, wirbt die Sammelbiene Neulinge an. Ein Teil des erhöhten Angebotes wird durch die Neulinge aufgefangen, jedoch wächst die Neulingszahl nicht proportional zur Erhöhung des Angebotes. Anwerbung im Stock und Wettbewerb zwischen den Sammelbienen an der Futterquelle schließen einen Regelkreis. Dadurch gleicht sich diese Sammelschar jederzeit dem veränderlichen Angebot an der Futterquelle an. Durch Änderung des Angebots an der Futterquelle lassen sich die Schwelle für das Duftmarkieren bestimmen und verschiedene Intensitätsstufen des Duftmarkierens unterscheiden. Bei konstantem Angebot an der Futterquelle lassen sich Schwankungen in der Dauer des Duftmarkierens und der Länge der Saugdauer bzw. der Größe der Abflugmagenfüllung ablesen, die gewisse Gesetzmäßigkeiten aufweisen. Mit Hilfe von Funktionsschemata werden Deutungen für diese Erscheinungen vorgeschlagen.     

Suprabenthic Peracarida (Crustacea, Malacostraca) sampled at 75°N off East Greenland

During an expedition with RV Polarstern in 1994 samples were taken at 75°N off Greenland, by means of an epibenthic sledge, which was equipped with an epi- and an additional suprabenthic net with an opening 1–1.33 m above the seafloor. Within the Peracarida (Crustacea, Malacostraca) sampled, 12 species were more frequent in the suprabenthic net, while 8 species were only sampled in this upper net. Received: 9 August 1996 / Accepted: 1 October 1996     

Reproductive Outcomes Following Ectopic Pregnancy: Register-Based Retrospective Cohort Study

Background

We aimed to compare reproductive outcomes following ectopic pregnancy (EP) versus livebirth, miscarriage, or termination in a first pregnancy.

Methods And Findings

A retrospective cohort study design was used. Scottish national data on all women whose first pregnancy occurred between 1981 and 2000 were linked to records of a subsequent pregnancy. The exposed cohort comprised women with an EP in their first pregnancy. There were three unexposed cohorts: women with livebirth, miscarriage, and termination of their first pregnancies. Any differences in rates of second pregnancy, livebirth, EP, miscarriage, or terminations and complications of a second ongoing pregnancy and delivery were assessed among the different exposure groups. A total of 2,969 women had an initial EP; 667,299 had a livebirth, 39,705 women miscarried, and 78,697 terminated their first pregnancies. Women with an initial EP had an increased chance of another pregnancy within 2 years (adjusted hazard ratio (AHR) 2.76 [95% CI 2.58–2.95]) or after 6 years (AHR 1.57 [95% CI 1.29–1.91]) compared to women with a livebirth. In comparison with women with an initial miscarriage, women who had an EP had a lower chance of a second pregnancy (AHR 0.53 [95% CI 0.50–0.56]). Compared to women with an initial termination, women with an EP had an increased chance of a second pregnancy (AHR 2.38 [95% CI 2.23–2.55]) within 2 years. Women with an initial EP suffered an increased risk of another EP compared to women with a livebirth (AHR 13.0 [95% CI 11.63–16.86]), miscarriage (AHR 6.07 [95% CI 4.83–7.62]), or termination (AHR 12.84 [95% CI 10.07–16.37]). Perinatal complications in a pregnancy following EP were not significantly higher than those in primigravidae or in women with a previous miscarriage or termination.

Conclusion

Women with an initial EP have a lower chance of conception than those who miscarry but an increased risk of a repeat EP in comparison with all three comparison groups. A major limitation of this study was the inability to separate women using contraception from those who were intending to conceive. Please see later in the article for the Editors'' Summary     

An atypical RNA pseudoknot stimulator and an upstream attenuation signal for -1 ribosomal frameshifting of SARS coronavirus

The −1 ribosomal frameshifting requires the existence of an in cis RNA slippery sequence and is promoted by a downstream stimulator RNA. An atypical RNA pseudoknot with an extra stem formed by complementary sequences within loop 2 of an H-type pseudoknot is characterized in the severe acute respiratory syndrome coronavirus (SARS CoV) genome. This pseudoknot can serve as an efficient stimulator for −1 frameshifting in vitro. Mutational analysis of the extra stem suggests frameshift efficiency can be modulated via manipulation of the secondary structure within the loop 2 of an infectious bronchitis virus-type pseudoknot. More importantly, an upstream RNA sequence separated by a linker 5′ to the slippery site is also identified to be capable of modulating the −1 frameshift efficiency. RNA sequence containing this attenuation element can downregulate −1 frameshifting promoted by an atypical pseudoknot of SARS CoV and two other pseudoknot stimulators. Furthermore, frameshift efficiency can be reduced to half in the presence of the attenuation signal in vivo. Therefore, this in cis RNA attenuator represents a novel negative determinant of general importance for the regulation of −1 frameshift efficiency, and is thus a potential antiviral target.     

Interference between cues of the same outcome in a non-causally framed scenario

Retroactive interference between cues of the same outcome (i.e., IbC) occurs when the behavioral expression of an association between a cue and an outcome (e.g., A → O1) is reduced due to the later acquisition of an association between a different cue and the same outcome (e.g., B → O1). Though this interference effect has been traditionally explained within an associative framework, there is recent evidence showing that IbC effect may be better understood in terms of the operation of higher order causal reasoning processes. The results from Experiments 1 and 2 showed an IbC effect in a learning task within a game scenario suggesting non-causal relationships between events. Thus, these results showed that IbC may have a diverse origin, one of them being of an associative nature.     

Biosynthesis of polymyxin E III. Total synthesis of polymyxin E by a cell-free enzyme system

Polymyxin E, an antimicrobial branched cyclic decapeptide, was synthesized by an enzyme fraction partially purified from crude extracts of the producing organism, $\text{Aerobacillus}\text{polyaerogenes}$. For the synthesis, three constituent amino acids (L-2,4-diaminobutyric acid, L-leucine, and L-threonine), ATP, Mg²⁺ and an acylating system consisting of octanoyl CoA and an ammonium sulfate fraction of cell extracts are required.     

The suitability of some plant hosts for the development of the peach-potato aphid, Myzus persicae (Sulzer)

Myzus persicae develop well on Brassica species, particularly those that grow rapidly, such as mustard and turnip, but much less well on sugar beet, spinach and lettuce. They usually develop better on young, rapidly growing leaves than on mature leaves. Two clones of M. persicae behaved similarly on the host plants tested.

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Zusammenfassung Die Entwicklung von Myzus persicae-Alienicolen auf den Sekundärwirten Kohlrüben, Kopfkohl, Chinakohl, Brokkoli, Blumenkohl, Senf, Raps, Grünkohl sowie Zuckerrübe, Spinat und Salat wurde durch wiederholte Erfassung der durchschnittlichen und totalen Larvenproducktion einzelner Jungfern sowie ihrer Mortalität in Gewächshausversuchen verfolgt und mit Auszählung des Befalls von Freilandpflanzen verglichen. Myzus persicae entwickelte sich gut an Brassica-Arten, besonders an raschwüchsigen wie Senf und Kohlrüben, aber viel weniger gut an Zuckerrübe, Spinat and Salat. Sie entwickelt sich gewöhnlich an jungen schnell wachsenden Blättern besser als an ausgereiften. Zwei Klone von Myzus persicae verhielten sich an den geprüften Wirtspflanzen ähnlich.

     

Light Scattering Investigation of Electric Field Alignment of Phospholipid Tubules

The polymerizable diacetylenic phospholipid 1,2-bis(10,12,tricosadiynoyl)-sn-glycero-3-phosphocholine (DC₂₃PC) forms straight hollow cylinders in water. Using an ac electric field it was possible to achieve significant orientational alignment of the tubules parallel to the field direction, and from light scattering results deduce an effective dielectric susceptibility anisotropy Δχ_E. Moreover, we suggest that the alignment arises from an orientational anisotropy of the total electrostatic enthalpy for a dielectric tubule in an electric field, rather than an inherent polarizability anisotropy of the constituent DC₂₃PC molecules, as was the case with magnetic field alignment.     

富平石川河的生态修复

自国家提出构建山水林田湖草生命共同体以来,已启动第三批山水林田湖草生态保护修复工程试点申报,但一些工程设计缺乏整体性,造成“真破坏”和“伪生态”的现象时有发生.本研究以石川河综合整治项目为例,针对石川河乱采乱挖河道砂石,工业污染,附近村民焚烧垃圾造成气、水和土污染极其严重,生态、人居环境持续恶化,鱼虾绝迹,污水横流,垃圾成堆等问题,采用重建生态河道、恢复生态、景观重塑、土地平整、PE复合土工膜、土壤修复等技术构建山水林田湖草生命共同体,并分析了生态、经济和社会效益,通过项目实施,为石川河流域提供了洁净的土壤环境、稳定的土体结构,有效改善了生态环境,提高了生活质量,实现生产发展、生态良好,不断满足社会发展建设美丽乡村的需要,同时为构建流域山水林田湖草生命共同体提供理论依据和实践支撑.     

An inclusive fitness analysis of synergistic interactions in structured populations

We study the evolution of a pair of competing behavioural alleles in a structured population when there are non-additive or ‘synergistic’ fitness effects. Under a form of weak selection and with a simple symmetry condition between a pair of competing alleles, Tarnita et al. provide a surprisingly simple condition for one allele to dominate the other. Their condition can be obtained from an analysis of a corresponding simpler model in which fitness effects are additive. Their result uses an average measure of selective advantage where the average is taken over the long-term—that is, over all possible allele frequencies—and this precludes consideration of any frequency dependence the allelic fitness might exhibit. However, in a considerable body of work with non-additive fitness effects—for example, hawk–dove and prisoner''s dilemma games—frequency dependence plays an essential role in the establishment of conditions for a stable allele-frequency equilibrium. Here, we present a frequency-dependent generalization of their result that provides an expression for allelic fitness at any given allele frequency p. We use an inclusive fitness approach and provide two examples for an infinite structured population. We illustrate our results with an analysis of the hawk–dove game.     

PGE2 induces the transition from non-adherent to adherent bone marrow mesenchymal precursor cells via a cAMP/EP2-mediated mechanism

When mesenchymal precursor cells from bone marrow are cultured in the presence of dexamethasone, the existence of distinct non-adherent and adherent populations can be demonstrated. The addition of PGE₂, forskolin, or dibutyryl-cAMP can induce a transition from the former to the latter and this may be an important mechanism in the bone anabolic effects of PGE₂. On the other hand, phorbol 12-myristate 13-acetate (PMA), an activator of protein kinase C, and sulprostone, an agonist for the PGE₂ receptor EP₁/EP₃ subtypes, had no effect. The phosphodiesterase inhibitor, isobutylmethylxanthine (IBMX), had a synergistic effect in combination with PGE₂, whereas neomycin, an inhibitor of inositol phosphate activity, had no effect, and LiCl, an inhibitor of inositol triphosphate metabolism, had an inhibitory effect on the PGE₂-induced transition. Consistent with this, the addition of PGE₂ to non-adherent bone marrow cells caused a 100% increase in cAMP synthesis. These results suggest that the induction of the transition from non-adherent to adherent osteoblast precursor is mediated by the EP₂-PGE₂ receptor subtype via an increase in intracellular cAMP synthesis.     

Immunogenic protein MPB57 from Mycobacterium bovis BCG: Molecular cloning, nucleotide sequence and expression

Using a single-probe method, we have cloned the gene for an immunogenic MPB57 protein of Mycobacterium bovis BCG. The nucleotide sequence includes an ORF of 300 base pairs encoding a protein of 99 amino acids with an M_r of 10 818. This cloned gene was expressed in an Escherichia coli expression vector to give a mature protein which reacted with a polyclonal antibody raised against MPB57.     

Eine neue Mikromethode zur Bestimmung der Lichtbrechung von Protoplasmatropfen

Zusammenfassung Eine bei Gebrauch einer Planparallelplatte und eines Uhrglases auf Ausmessungen an Fizeauschen Interferenzkurven beruhende, an mikroskopische Grö\enverhÄltnisse angepa\te Methode des Physikers wird nach Prinzip, Durchführung und Anwendbarkeitsgrenzen behandelt. Es werden vorlÄufige Ergebnisse an Modellflüssigkeiten und Protoplasmatropfen mitgeteilt.     

Tensile Mechanics of Alanine-Based Helical Polypeptide: Force Spectroscopy versus Computer Simulations

In nature, an α-helix is commonly used to build thermodynamically stable and mechanically rigid protein conformations. In view of growing interest in the mechanical rigidity of proteins, we measured the tensile profile of an alanine-based α-helical polypeptide on an atomic-force microscope to investigate the basic mechanics of helix extension with minimal interference from side-chain interactions. The peptide was extended to its maximum contour length with much less force than in reported cases of poly-L-Glu or poly-L-Lys, indicating that chain stiffness strongly depended on the physicochemical properties of side chains, such as their bulkiness. The low tensile-force extension originated presumably in locally unfolded parts because of spontaneous structural fluctuations. In 50% trifluoroethanol, the well-known helix-promoting agent, the rigidity of the sample polypeptide was markedly increased. Computer simulations of the peptide-stretching process showed that a majority of constituent residues underwent a transition from an α-helical to an extended conformation by overcoming an energy barrier around ψ ∼0° on the Ramachandran plot. The observed lability of an isolated helix signified the biological importance of the lateral bundling of helices to maintain a rigid protein structure.     

Evidence that N-methyl-N′-nitro-N-nitrosoguanidine induces adaptive response in Bacillus thuringiensis

Bacillus thuringiensis is shown to have an inducible error-free repair system for alkylation damage as found in Escherichia coli and Bacillus subtilis. Growth of cells in the presence of low concentrations of N-methyl-N′-nitro-N-nitrosoguanidine (MNNG) induces an adaptive response which is characterized by an increase in resistance to killing and mutagenesis by challenge with higher concentrations of MNNG. In addition, we have noted with interest that adaptive low doses seem to produce lesions at a rate sufficient to induce an increase of mutation frequency, and inhibition of cell division. The possibility of an interaction between SOS and adaptive responses with these low doses of MNNG is discussed.