In Vitro Studies of α-Carboxyl-3-Thienylmethyl Penicillin,a New Semisynthetic Penicillin

The activity of a new semisynthetic penicillin, α-carboxyl-3-thienylmethyl penicillin (BRL-2288) was determined against 535 clinical isolates of gram-negative bacilli, by using the tube dilution technique. Nearly 80% of isolates of Proteus spp. were inhibited by 3.12 μg or less of this antibiotic per ml. BRL-2288 was as active as ampicillin against Escherichia coli. It was slightly more active than carbenicillin or 6-(d-α-sulfoaminophenylacetamido)-penicillanic acid against Pseudomonas sp., with over half of the isolates being inhibited by 50 μg or less of BRL-2288 per ml. Isolates of Klebsiella sp. were routinely resistant to this antibiotic. The drug was bactericidal against most sensitive organisms. BRL-2288 was less active against large inocula. A strain of Pseudomonas sp. which developed resistance to carbenicillin also developed resistance to BRL-2288 simultaneously.     

Effect of β-Lactamase Location in Escherichia coli on Penicillin Synergy

Resistance to ampicillin in Escherichia coli is due generally to the presence of a beta-lactamase (penicillinase). Resistant strains have been found to fall into two groups: those with high-level resistance (1,000 mug/ml or greater) and those with low-level resistance (8 to 250 mug/ml). Most of the high-level resistant organisms posses beta-lactamases whose synthesis is episomally mediated. These strains release penicillinase from the cell when they are subjected to osmotic shock. Low-level resistant strains do not release the enzyme with osmotic shock. High-level resistant strains are not susceptible to the synergistic action of a penicillinase-resistant penicillin with ampicillin. Seventy eight per cent of low-level resistant strains are susceptible to the synergistic action of ampicillin and oxacillin. The two types of beta-lactamases are similar in regard to most properties; both enzymes are subject to competitive inhibition by penicillinase-resistant penicillins. The difference in location in the cell might explain why only some strains of E. coli are susceptible to the synergistic action of penicillin combinations.     

Some Problems Involved in Ampicillin Formation by Kluyvera’s Penicillin Acylase

The cell growth of Kluyvera citrophila KY3641, capable of producing α-aminobenzyl-penicillin (APc) from 6-aminopenicillanic acid (6-APA) and phenylglycine, was stimulated by glutamic acid, serine or proline, or by pH control with tartaric acid or fumaric acid.

Penicillinase produced in an early stage of growth or pH-controlled culture was inactivated by alkaline treatment (incubation of cells at 40°C for 5 to 24 hr in pH 7.5 to 9.5) without inactivation of penicillin acylase. Surface active agents enhanced APc production. On the other hand, phenylalanine and some inorganic compounds inhibited this production.

This bacterium formed APc from penicillin G, but amounts of APc formed were only 9 μg from 20 mg of penicillin G.     

Veränderungen des elektrokinetischen Potentials bei der Adaptation der Staphylokokken an Penicillin

Ohne ZusammenfassungInstitut für Mikrobiologie u. Serologie und Institut für Biochemie     

Penicillin acylase immobilization depending on macromolecular crowding and catalysis in aqueous–organic medium

To enhance penicillin acylase (PA) performance, it was immobilized in mesocellular silica foams (MCFs) depended on macromolecular crowding and applied to catalysis in non-aqueous medium. Ficoll 70, dextran 10,000, dextran 40,000 and bovine serum albumin were co-assembled with PA. It was observed that specific activity of PA assembled in MCFs with dextran 10,000 in 80% cyclohexane (v/v) was 233.2 U/mg, 200% as that of PA assembled in MCFs in 80% cyclohexane and 323.5% as that of free PA in full aqueous medium. As content of alkane increased, activity of PA in MCFs with macromolecules varied slightly. In addition, PA co-immobilized with dextran 10 in MCFs retained 58.2% of its initial activity after heating at 50°C for 4 h, 1.2 times higher than that of PA immobilized alone in MCFs. The results showed that macromolecular crowding was favorable for immobilization of PA and its catalysis in suitable aqueous–organic medium.     

In Vitro Studies of a New Semisynthetic Penicillin, 6-(D-α-Sulfoaminophenylacetamido)-Penicillanic Acid

The activity of 6-(D-α-sulfoaminophenylacetamido)-penicillanic acid was determined against 357 clinical isolates of gram-negative bacilli by use of the tube-dilution technique. The majority of the isolates of Pseudomonas species were inhibited by 200 μg/ml or less of this antibiotic. Most of the isolates of Escherichia coli had a minimal inhibitory concentration of 50 μg/ml or less. Seventy-three per cent of the isolates of P. mirabilis, 40% of the isolates of P. morganii, and 45% of the isolates of Enterobacter species were inhibited by 12.5 μg/ml or less, whereas most of the isolates of Klebsiella species and Serratia species were resistant. The activity of this semisynthetic penicillin was affected by the size of the inoculum. The drug was bactericidal against all isolates of E. coli and Proteus species that were sensitive to it, but it was bactericidal against only 32% of the sensitive isolates of Pseudomonas species.     

Direct modulatory effect of hexasodium N,N,N′,N′-ethylenediamine-tetramethylenephosphonate on bone cell function in vitro

The phosphonate hexasodium N,N,N′,N′-ethylenediamine-tetramethylenephosphonate (EDITEMP · Na₆) reduced alkaline phosphatase (Alp) activity and cAMP response to parathyroid hormone (PTH) in primary cultures of foetal rat calvaria cells in a dose-dependent manner, while not affecting culture DNA content. EDITEMP · Na₆ also inhibited the mineralization of three-dimensional bone nodules formed in vitro, but not the number of nodules formed. Bone cell culture DNA content was also reduced by EDITEMP · Na₆ but at concentrations in excess of those needed to modulate osteoblastic cell function. Withdrawal of EDITEMP · Na₆ led to slow but complete recovery of Alp activity. At EDITEMP · Na₆ concentrations of 25 μM and higher, recovery of Alp activity appeared to be independent of protein and/or DNA synthesis. Cell culture acid phosphatase (Acp) activity was not affected by EDITEMP · Na₆. The results indicate that EDITEMP · Na₆ has a direct inhibitory effect on (mature) osteoblastic cell function. In the presence of bone tissue this inhibition also occurred, although not at a relatively low dose level.     

Fluorescence and circular-dichroism studies on the Streptomyces R61 dd-carboxypeptidase–transpeptidase. Penicillin binding by the enzyme

The circular dichroism of the dd-carboxypeptidase-transpeptidase from Streptomyces R61 shows in the near u.v. a set of weak extrema at 289nm (positive) and at 282, 275 and 268nm (all negative). In the far u.v. it shows negative extrema at 217-218 and 208nm, crossover at 202nm and a positive maximum at about 194nm. The u.v. absorption of the enzyme shows it to contain tyrosine and tryptophan in approx. 3.4:1 ratio. The enzyme is fluorescent with a maximum emission at 318-320nm. The near-u.v. circular dichroism of the protein is extensively affected by binding of penicillin G, but the far u.v. is unaffected. Binding of the antibiotic also causes quenching of the fluorescence of the enzyme. The latter effect has been used to study the binding of penicillin G to the enzyme and the influence exerted upon it by salts, denaturants and peptide substrates and inhibitors. High-affinity binding of penicillin appears to be comparatively slow and reversible, and can occur under conditions in which the protein is enzymically inactive. The thermal denaturation of the enzyme in guanidinium chloride at pH7 is affected by binding of the antibiotic. The presence of even large concentrations of beta-mercaptoethanol neither impaired the activity of the enzyme nor prevented its inhibition by penicillin G or cephalosporin C. A new hypothesis for the molecular mechanism of the interaction of the enzyme with penicillin is proposed.     

β-Lactamase-Free Penicillin Amidase from Alcaligenes sp.: Isolation Strategy, Strain Characteristics, and Enzyme Immobilization

Isolation and characterization of a β-lactamase (EC 3.5.2.6)-free, penicillin amidase (penicillin amidohydrolase, EC 3.5.1.11)-producing organism is reported. The test strain was isolated by an enrichment technique with a substrate other than penicillins. The isolated strain belongs to the genus Alcaligenes. Phenylacetic acid was found to be the inducer of penicillin amidase. The amidase has a broad substrate spectrum. It is very active against penicillin G and semisynthetic cephalosporins, whereas penicillin V and semisynthetic penicillins acted moderately as a substrate. Immobilized cells of Alcaligenes sp. were shown to act as a reversible enzyme. Received: 28 April 1999 / Accepted: 18 May 1999     

Fluorinated N,N′-diarylureas as AMPK activators

Adenosine monophosphate-activated kinase (AMPK) plays a central role in regulating energy homeostasis in eukaryotic cells. AMPK also regulates lipid synthesis by inhibiting acetyl-CoA carboxylase (ACC) and regulates mTOR signaling by activating TSC2. Due to its important roles in cell metabolism, AMPK is an attractive target for metabolic diseases, such as type II diabetes and obesity. AMPK activators, such as metformin, that are used for diabetes treatment are also effective anticancer agents. However, the efficacies of many known AMPK activators are relatively low. For example, metformin activates AMPK at millimolar levels. In this study, we identified a novel family of AMPK activators, namely fluorinated N,N′-diarylureas, that activate AMPK at 1–3 μM concentrations. These novel agents strongly inhibit the proliferation of colon cancer cells. We studied the potential mechanisms of these agents, performed a structure–activity relationship (SAR) study and identified several fluorinated N,N′-diarylureas as potent AMPK activators.     

N的影响

植物和土壤中的¹⁵N自然丰度值(δ¹⁵N)是评价生态系统N循环的一个重要指标, 而放牧是草原生态系统的主要土地利用方式, 对草原生态系统的N循环过程的改变起着重要作用。该研究测定了内蒙古锡林河流域放牧和围封条件下草原群落主要优势植物和土壤的δ¹⁵N值, 探讨放牧对草原N循环的影响。研究中所测定的8种植物叶片δ¹⁵N变化很大(–4.04‰–4.34‰), 但与植物功能型有一定的相关性。放牧显著降低了大针茅(Stipa grandis)、杂类草和小半灌木木地肤(Kochia prostrata)的δ¹⁵N值。具有潜在共生固氮能力的豆科植物δ¹⁵N偏低负值(–4.04‰ – –1.90‰), 但在放牧和围封条件下无显著差异; 而被认为具有联合固氮能力的羊草(Leymus chinensis), 放牧后δ¹⁵N显著增加, 一定程度上表明了豆科植物和羊草生物固氮能力的存在。所有植物中, 除无菌根侵染的木地肤外, 其他有丛枝菌根真菌侵染记录的物种δ¹⁵N值较低, 通常接近0或为负值, 说明在N限制的内蒙古草原, 菌根转运N可能也是一种重要的N源途径。放牧显著降低了0–20 cm土壤δ¹⁵N值, 这也与过去的研究结果不同。δ¹⁵N的测定为生态系统提供了一个整合时空N循环过程的综合指标, 反映出放牧改变了草原生态系统的N循环。     

Stereoselective and driving-force-dependent photoinduced electron-transfer reactions of zinc myoglobin with optically active N,N′-dimethylcinchoninium and N,N′-dimethylcinchonidinium ions

In order to understand the detailed mechanism of the stereoselective photoinduced electron-transfer (ET) reactions of zinc-substituted myoglobin (ZnMb) with optically active molecules by flash photolysis, we designed and prepared new optically active agents, such as N,N′-dimethylcinchoninium diiodide ([MCN]I₂) and N,N′-dimethylcinchonidinium diiodide ([MCD]I₂). The photoexcited triplet state of ZnMb, ³(ZnMb)*, was successfully quenched by [MCN]²⁺ and [MCD]²⁺ ions to form the radical pair of ZnMb cation (ZnMb^·+) and reduced [MCN]^·+ and [MCD]^·+, followed by a thermal back ET reaction to the ground state. The rate constants (k _q) for the ET quenching at 25 °C were obtained as k _q(MCN)=(1.9±0.1)×10⁶ M⁻¹ s⁻¹ and k _q(MCD)=(3.0±0.2)×10⁶ M⁻¹ s⁻¹, respectively. The ratio of k _q(MCD)/k _q(MCN)=1.6 indicates that the [MCD]²⁺ preferentially quenches ³(ZnMb)*. The second-order rate constants (k _b) for the thermal back ET reaction from [MCN]^·+ and [MCD]^·+ to ZnMb^·+ at 25 °C were k _b(MCN)=(0.79±0.04)×10⁸ M⁻¹ s⁻¹ and k _b(MCD)=(1.0±0.1)×10⁸ M⁻¹ s⁻¹, respectively, and the selectivity was k _q(MCD)/k _q(MCN)=1.3. Both quenching and thermal back ET reactions are controlled by the ET step. In the quenching reaction, the energy differences of ΔΔH ^≠(MCD–MCN) and ΔΔS ^≠(MCD–MCN) at 25 °C were obtained as −1.1 and 0 kJ mol⁻¹, respectively. On the other hand, ΔΔH ^≠(MCD–MCN)=11±2 kJ mol⁻¹ and TΔΔS ^≠(MCD–MCN)=−10±2 kJ mol⁻¹ were given in the thermal back ET reaction. The highest stereoselectivity of 1.7 for [MCD]^·+ found at low temperature (10 °C) was due to the ΔΔS ^≠ value obtained in the thermal back ET reaction. Electronic Supplementary Material Supplementary material is available for this article at and is accessible for authorized users.     

Bioaccumulation and Biovolatilisation of Pentavalent Arsenic by Penicillin janthinellum, Fusarium oxysporum and Trichoderma asperellum Under Laboratory Conditions

Some fungi are able to control and remediate arsenic (As)-contaminated soil, sediment, or water. Here, we investigate potential accumulation and volatilisation of As by three fungi strains. Results indicated that the highest level of As was accumulated by Penicillin janthinellum with 39.54 μg after 10 days in the culture system amended with 2,500 μg As(V), which represents 50 mg/l As. Fusarium oxysporum showed the highest amount of volatilised As with 304.06 μg after 15 days. The As content in the treated system (filter paper + As + fungi) was significantly higher than that in the control (filter paper + As; filter paper + fungi; filter paper). Trichoderma asperellum and F. oxysporum showed superior abilities for the absorption of extracellular As and accumulation of intracellular As, which accounted for 82.2 and 63.4% of the total accumulated As, respectively. However, P. janthinellum presented an equal distribution of intracellular and extracellular As. Scanning electron microscope (SEM) analysis suggested that little impact on mycelium growth of the three fungal strains was seen after exposure to 50 mg/l As(V) for 5 days, while the growth of fungi in the control was inhibited. The present results demonstrate that P. janthinellum, F. oxysporum, and T. asperellum would be expected to tackle As-contaminated environments.     

The Structure of “N1, N6-Carbonyladenosine”

Abstract

Re-interpretation of the available data led to structural assignment of the title N¹, N⁶carbonyladenosine (1b) as N⁶,N⁶-carbonyldiadenosine (4b).     

应用N端Cys-free断裂蛋白质内含子标记蛋白质N端

蛋白质的特异位点修饰可以帮助了解蛋白质的结构与功能.但是,现有的蛋白质特异位点标记方法种类有限,而且存在局限性,所以有必要开发新的蛋白质特异位点标记方法．以谷胱甘肽-S-转移酶(GST)为研究对象,借助蛋白质反式剪接技术,建立了利用新型断裂蛋白质内含子对蛋白质进行N 端标记的新方法.在这个方法中,通过简单的重组表达、标记和纯化得到带有荧光基团的小肽,经过蛋白质反式剪接,荧光基团被标记到蛋白质的N 端. 初步研究结果显示,标记效率可达到12 %.     

人禽流感H7N9、H5N1、H10N8临床特征

人禽流感(human-avian influenza)是一种由禽流感病毒中某些亚型感染人所引起的急性呼吸道传染病,目前能够感染人的禽流感病毒主要有H5、H7、H9和H10亚型。人感染禽流感病毒A(H5N1)、A(H7N9)、A(H10N8)与重症季节性流感临床表现相似,主要表现为重症肺炎,病死率高。人禽流感与重症季节性流感和重症新甲型H1N1流感患者有相同的危险因素,如高龄、合并基础疾病等,但人禽流感临床表现更重。     

Antimicrobial efficacy of some N,N’-Dialkyl-N,N’-dimethyl-1, 6-hexanediamine dioxides

A total of 17 N,N'-dialkyl-N,N'-dimethyl-1,6-hexanediamine dioxides were tested for activity against three microorganisms. A relationship was found between the length of the alkyl substituent and antimicrobial activity.