Autophagosomes,phagosomes, autolysosomes,phagolysosomes, autophagolysosomes… Wait,I’m confused

When an autophagosome or an amphisome fuse with a lysosome, the resulting compartment is referred to as an autolysosome. Some people writing papers on the topic of autophagy use the terms “autolysosome” and “autophagolysosome” interchangeably. We contend that these words should be used to denote 2 different compartments, and that it is worthwhile maintaining this distinction—the autophagolysosome has a particular origin in the process of xenophagy that makes it distinct from an autolysosome.     

Autophagosomes,phagosomes, autolysosomes,phagolysosomes, autophagolysosomes… Wait,I’m confused

When an autophagosome or an amphisome fuse with a lysosome, the resulting compartment is referred to as an autolysosome. Some people writing papers on the topic of autophagy use the terms “autolysosome” and “autophagolysosome” interchangeably. We contend that these words should be used to denote 2 different compartments, and that it is worthwhile maintaining this distinction—the autophagolysosome has a particular origin in the process of xenophagy that makes it distinct from an autolysosome.     

Der Mengen- und Spurenelementgehalt des Rinderhaares als Indikator der Calcium-, Magnesium-, Phosphor-, Kalium-, Natrium-, Eisen-, Zink-, Mangan-, Kupfer-, Molybdän- und Kobaltversorgung

Many health effects can be attributed to the Mediterranean herb oregano (Origanum vulgare L.) and several studies demonstrated the improving effect on performance, changes in blood count, antibacterial, antifungal and immunmodulating abilities. The majority of these investigations were carried out with processed essential oil, while whole plant material was only used in a few studies. Thus, the aim of the present experiment was to test the effect of increasing proportions of dried oregano in piglet feed on health and performance, with a special focus on immune modulation. A total of 80 male castrated weaned piglets (body weight [BW] 7.9 kg ±1.0 kg) were used in a feeding experiment lasting 5 weeks. They were assigned to 4 experimental groups: a control diet, and three diets with an oregano supplementation at 2 g, 4 g and 8 g per kg feed, respectively, corresponding to 23.5 mg, 46.9 mg and 93.9 mg carvacrol/kg DM. After 3 weeks, half of each group was challenged with 5 µg lipopolysaccharides (LPS) per kg BW. Blood samples were collected 2 h after LPS stimulation and analysed for T-cell phenotypes, granulocyte activity, clinical-chemistry as well as white and red blood count. The results indicate no effects of oregano on performance. In contrast, oregano altered the lymphocyte proportion and the ratio of CD4⁺ and CD8⁺ T-cells as well as the triglyceride concentration in the serum of non-stimulated and in LPS-stimulated piglets. In conclusion, whole plant supplementation of oregano to piglet feed altered immune-related parameters, but did not modulate the acute inflammatory response induced by LPS stimulation.     

Der Einfluß endogener Faktoren auf den Asche-, Kalzium-, Magnesium-, Kalium-, Natrium-, Phosphor-, Zink-, Eisen-, Kupfer- und Mangangehalt der Schweineborste

The objective of the present study was to investigate physiological effects of a marginal copper and iron supply on pigs. Therefore an experiment was conducted with 4 × 12 growing pigs of the crossbreed Pietrain × Deutsche Landrasse. The animals were fed for a period of 119 days with a diet poor of copper (1.5 mg Cu/kg diet) and/or poor of iron (35 mg Fe/kg diet). Control animals were supplied adequately with copper (4.8 mg Cu/kg diet) and iron (85 mg Fe/kg diet). The diet was given according to weight. After reaching an average weight of 102.6 ± 3.5 kg the animals were slaughtered. Due to the activity of the coerulplasmin and katalase enzyme and the haematological parameters, the supply of copper and iron could be classified as marginal. There was no interaction between copper deficiency and iron metabolism. The protein metabolism was unchanged. Low copper intake reduced the copper concentrations in serum, liver, muscle and backfat, and low iron intake reduced the iron concentration in serum, liver and muscle. Marginal copper and iron supply had no relevant effect on either food intake and growth performance or carcass characteristics and meat quality.     

Der Mengen- und Spurenelementgehalt des Rinderhaares als Indikator der Calcium-, Magnesium-, Phosphor-, Kalium-, Natrium-, Eisen-, Zink-, Mangan-, Kupfer-, Molybdän- und Kobaltversorgung

Beyond the energy requirement of maintenance, the assimilated energy, occurring in bioproducts, is linearly proportional to the intake of metabolizable energy in non‐underfed conditions. In contrast, resting metabolic rate is differing between individuals within a population of an animal species. As adaptability to changed environmental conditions may play a role, young bulls were exposed to thermoneutral (18°C) and low (4°C) ambient temperatures and were fed at two feeding levels (1.0 and 1.6 times energy requirement in maintenance) to produce metabolic rate differences, using the same animals, metabolic rate was altered by reducing the sympathetic outflow in each case. Expression of sulfonylurea receptors in circulating mononuclear leukocytes and cells from skeletal muscle (m. semitendinosus) was studied by flow cytom‐etry. Changes of metabolic rate at rest corresponded to the portion of cells with sulfonylurea receptors expression. The data from reducing the sympathetic outflow and those from sulfonylurea receptors expression are useful to explain metabolic rate differences among individuals of an animal population.     

胸怀祖国, 忘我工作, 为国争光

刘以训教授是我国著名的生物学家,中国科学院院士。他致力于生殖生物学研究,在排卵、黄体萎缩、精子发生和胚胎植入等系列研究上,成绩卓著,享有国际声誉。读罢此文,钦敬之情油然而生。     

Barley—Botany,production, harvesting,processing, utilization and economics

Barley, probably the oldest cultivated cereal, is widely grown in cooler areas of the world. The annual world production of nearly two and a half billion bushels exceeds that of rye but is less than that of rice, wheat, corn and oats, respectively. Most of the annual 300 million dollar crop of the U.S. is fed to livestock, but about one- third is manufactured into malt.     

人口,食物,资源,环境与生物学

人口、食物、资源、环境与生物学钱燕文（中国科学院动物研究所北京１０００８０）人家都说下一个世纪是生物学的世纪,人口、食物、资源、环境也是近２０年来的热门话题,因为这四个方面关系到我们人类的未来。为什么下个世纪是生物学的世纪？也得从人口、食物、资源、环...     

基因工程人胰岛素原和胰岛素的分离纯化及性质研究

E. coli DH 5 alpha cells harboring a plasmid pWR 590-BCA 4 for fused human proinsulin production were cultured. The fused human proinsulin was isolated from the fermented cells and then subjected it to cleavage with BrCN. The cleaved product was then converted to crude proinsulin-S-sulfonate using oxidative sulfitolysis. The isolation of human proinsulin-S-sulfonate was accomplished by ion exchange chromatography on QAE-sephadex A-25, followed by gel filtration on sephadex G-50. The purified human proinsulin-S-sulfonate was folded using a disulfide interchange method. The folding mixture was then chromatographed on sephadex G-50 and purified proinsulin was obtained. The proinsulin was then converted to human insulin and C-peptide by a combination cleavage with trypsin and carboxypeptidase B. The total yield of human insulin was about 5 mg/L The Zinc insulin crystals were obtained with amorphous human insulin using citrate method. The amino acid composition N-terminal sequences as well as C-terminal amino acid residues are in agreement with expected results. The hypoglycemic activity of purified human insulin is 26-27 U/mg, as judged by mouse convulsion assay, and the RIA activity is about 99% of that of porcine insulin.     

Fibrillation of human insulin A and B chains

Human insulin, which consists of disulfide cross-linked A and B polypeptide chains, readily forms amyloid fibrils under slightly destabilizing conditions. We examined whether the isolated A and B chain peptides of human insulin would form fibrils at neutral and acidic pH. Although insulin exhibits a pH-dependent lag phase in fibrillation, the A chain formed fibrils without a lag at both pHs. In contrast, the B chain exhibited complex concentration-dependent fibrillation behavior at acidic pH. At higher concentrations, e.g., >0.2 mg/mL, the B chains preferentially and rapidly formed stable protofilaments rather than mature fibrils upon incubation at 37 degrees C. Surprisingly, these protofilaments did not convert into mature fibrils. At lower B chain concentrations, however, mature fibrils were formed. The explanation for the concentration dependence of B chain fibrillation is as follows. The B chains exist as soluble oligomers at acidic pH, have a beta-sheet rich conformation as determined by CD, and bind ANS strongly, and these oligomers rapidly form dead-end protofilaments. However, under conditions in which the B chain monomer is present, such as low B chain concentration (<0.2 mg/mL) or in the presence of low concentrations of GuHCl, which dissociates the soluble oligomers, mature fibrils were formed. Thus, both A and B chain peptides can form amyloid fibrils, and both are likely to be involved in the interactions leading to the fibrillation of intact insulin.     

人胰岛素原类似物(BKRA)基因的合成与表达

为了利用基因工程生产胰岛素,按照已知的人胰岛素Ａ、Ｂ链氨基酸序列和大肠杆菌偏爱的氨基酸密码子设计并合成了人胰岛素原类似物（ＢＫＲＡ）基因,其中以赖（Ｋ）－精（Ｒ）二肽编码区取代人胰岛素原Ｃ肽编码区．为了避免其编码蛋白在大肠杆菌中表达时被降解,通过人工接头将２个ＢＫＲＡ基因串联起来,接头部分氨基酸序列为Ａｒｇ－Ａｒｇ－Ａｓｎ－Ｓｅｒ．将串联的ＢＫＲＡ基因克隆到表达载体ｐＥＴ－２８ａ（＋）,实现了在大肠杆菌中的融合表达,表达产物以包含体形式存在,约占细菌总蛋白２４％．表达产物氨基末端具有六组氨酸肽段,以ＨｉＴｒａｐ凝胶进行亲和层析,一步纯化可达纯度９５％以上．放射免疫测定表明,纯化的融合蛋白具有胰岛素抗原活性．表明已构建成人胰岛素原类似物的高效表达菌株     

Construction of a family of artificial genes of human insulin

Using oligonucleotide-directed mutagenesis and chemical-enzymatic DNA synthesis, genes for A and B insulin chains, C-peptide and Tyr-C-peptide have been constructed starting from synthetic gene for human proinsulin synthesized earlier. The genes for human preproinsulin, mini-proinsulin, single-chain insulin and their modifications were also synthesized. The constructions obtained were cloned in plasmid vectors.     

Influence of imprinting with A and B chains of insulin on binding and functional changes in tetrahymena

Insulin and its A and B chain increased the quantity of intracellular PAS-positive material (glycogen) in tetrahymena, whereas the combined A+B chains decreased it. Imprinting—previous interaction—with insulin, its A and B chains in themselves and with the A+B chain increased the hormone binding capacity of tetrahymena, but the functional effect of imprinting (storage or breakdown of glycogen) showed a different tendency with insulin and A+B chain on the one hand, and A chain and B chain on the other. Since the imprinting potential of a molecule promotes the induction of receptor formation, the fact remains that both component chains of insulin were able to act as potential imprinters, although the A chain was superior to the B chain in this respect throughout, and combined treatment with the A+B chain ultimately induced the formation of a similar binding site as insulin itself.     

反相高效液相层析在胰岛素结构与功能研究中的应用

采用一种简单的“甲醇-水-0.1％三氟醋酸”作为洗脱体系的反相高效液相层析对研究胰岛素结构与功能关系,以及A链和B链相互作用时所用的一些胰岛素衍生物进行了快速、灵敏、准确的分析和鉴定。 胰岛素的S-磺酸型A链和B链只经一次离子交换层析纯化,经鉴定结果表明是均一的。S-硫甲基型A链和B链基本上也是均一的。通过对胰岛素还原产物的分析确定了能保证胰岛素折分完全并对其重组较为有利的还原条件。去B链羧端五肽(B26—30)胰岛素,以及去B链羧端八肽(B23—30)胰岛素能同时与天然胰岛素清楚地分离。一级结构差别极小的猪胰岛素和牛胰岛素也可得到分辨。胰岛素的羧端缩短的B链和A链重组的定量分析表明B链羧端肽段不但对胰岛素分子的生物活力的体现,而且在A链和B链的正确重组中都起着重要的作用。     

The pairing of the separated A and B chains of insulin and its derivatives, FTIR studies.

From the amide I bands of their deconvolved FTIR spectra, the S-thiomethyl derivatives of the insulin A, B, despentapeptide(26-30) B and desoctapeptide(23-30) B chains all contain significant amounts of ordered secondary structure. The intact B chain is considerably more ordered than either the A or the truncated B chains. Comparison of the spectra of the separated and mixed intact chains of insulin suggests further folding upon mixing of the chains leading to significant increases in ordered secondary structures, presumably because of stabilization by interaction of the chains. The interactions of the A chain with the DPI B chain appear to be weaker as compared to that with the intact B chain. The above results suggest that only the intact A and B chains contain sufficient structural information to recognize each other and interact to form a native-like structure which make the correct formation of the disulfide linkages possible.     

Plasmid partition system of the P1par family from the pWR100 virulence plasmid of Shigella flexneri

P1par family members promote the active segregation of a variety of plasmids and plasmid prophages in gram-negative bacteria. Each has genes for ParA and ParB proteins, followed by a parS partition site. The large virulence plasmid pWR100 of Shigella flexneri contains a new P1par family member: pWR100par. Although typical parA and parB genes are present, the putative pWR100parS site is atypical in sequence and organization. However, pWR100parS promoted accurate plasmid partition in Escherichia coli when the pWR100 Par proteins were supplied. Unique BoxB hexamer motifs within parS define species specificities among previously described family members. Although substantially different from P1parS from the P1 plasmid prophage of E. coli, pWR100parS has the same BoxB sequence. As predicted, the species specificity of the two types proved identical. They also shared partition-mediated incompatibility, consistent with the proposed mechanistic link between incompatibility and species specificity. Among several informative sequence differences between pWR100parS and P1parS is the presence of a 21-bp insert at the center of the pWR100parS site. Deletion of this insert left much of the parS activity intact. Tolerance of central inserts with integral numbers of helical DNA turns reflects the critical topology of these sites, which are bent by binding the host IHF protein.     

还原型胰岛素A、B链相互作用的初步研究

本文报道在chance所用“一步法”重组胰岛素的类似条件下,还原型A、B链相互作用研究的初步结果。在pH10.6对S—磺酸型A及B链加入1.2倍SH/SSO_3—的二巯基苏糖醇,在一小时内A、B链与DTT的混合物较之单独的A、B链与DTT的混合物,紫外差光谱显示295、245nm负差峰,峰值随时间增大;园二色性测定观察到混合链的α螺旋含量比单独的A、B链有所增加。这表明在A、B链的巯基氧化生成二硫链前,还原型A、B链间的相互作用导致Tyr残基逐渐内埋,肽链构象的调整使有序二级结构增加。根据此时还原程度的测定,讨论了DTT与S—磺酸型A、B链作巯基交换的可能机制,过多的还原试剂易破坏已正确配对的二硫键,也妨碍肽链构象的调整。     

Plasmid DNAs encoding insulin and glutamic acid decarboxylase 65 have distinct effects on the progression of autoimmune diabetes in nonobese diabetic mice.

We previously demonstrated that administration of plasmid DNAs (pDNAs) encoding IL-4 and a fragment of glutamic acid decarboxylase 65 (GAD65) fused to IgGFc induces GAD65-specific Th2 cells and prevents insulin-dependent diabetes mellitus (IDDM) in nonobese diabetic (NOD) mice. To assess the general applicability of pDNA vaccination to mediate Ag-specific immune deviation, we examined the immunotherapeutic efficacy of recombinants encoding murine insulin A and B chains fused to IgGFc. Insulin was chosen based on studies demonstrating that administration of insulin or insulin B chain by a variety of strategies prevents IDDM in NOD mice. Surprisingly, young NOD mice receiving i.m. injections of pDNA encoding insulin B chain-IgGFc with or without IL-4 exhibited an accelerated progression of insulitis and developed early diabetes. Exacerbation of IDDM correlated with an increased frequency of IFN-gamma-secreting CD4(+) and CD8(+) T cells in response to insulin B chain-specific peptides compared with untreated mice. In contrast, treatment with pDNAs encoding insulin A chain-IgGFc and IL-4 elicited a low frequency of IL-4-secreting Th cells and had no effect on the progression of IDDM. Vaccination with pDNAs encoding GAD65-IgGFc and IL-4, however, prevented IDDM. These results demonstrate that insulin- and GAD65-specific T cell reactivity induced by pDNA vaccination has distinct effects on the progression of IDDM.