贝伐珠单抗的疗效预测研究现状

抗血管生成的贝伐珠单抗已被广泛应用于多种恶性肿瘤的治疗。但因其受益人群有限及逐渐出现的获得性耐药,也限制了其对患者总生存的改善。迫切需要寻找有效的预测贝伐珠单抗疗效的预测因子指导临床应用。人们在生物标记物、影像学方面进行了大量探索,但目前尚没有一个公认的普遍适用的临床预测指标,仍需要从多角度出发,进行进一步的基础研究及临床研究去发现和验证有效的疗效预测指标。     

Cover Image

The myodural bridge is represented by skeletal muscle fibers attaching to the cervical dura mater. It has been described from a variety of mammals and other amniotes. In this issue of the Journal of Morphology (pp. 123-133), Grondel and coauthors test an earlier assumption about the presence of the myodural bridge in snakes using histology, computed tomography, and micro-CT imaging. They find no evidence for a myodural ridge in snakes and discuss the functional implications. The cover image shows a transverse histological section through atlas and axis from Dasypeltis scabra.     

Catalase Test as an Aid to the Identification of Enterobacteriaceae

Although the catalase test has been used for many years for rapid differentiation of the genera of gram-positive organisms, little has been said about its use in the family Enterobacteriaceae. It was further noted that a wide variety of methods exist for the execution of the catalase test, that there is no universally accepted strength specified for the hydrogen peroxide, and that no gradations for the vigor and speed of the reaction have been mentioned. Under the conditions of the clinical laboratory, we have developed a simple, rapid, and accurate method for the catalase test that has been of great value as an aid in the identification of the Enterobacteriaceae. With 3% H(2)O(2), it was observed that Serratia, Proteus, and Providencia were vigorous catalase reactors. Only Salmonella and rare Escherichia, Enterobacter, and Klebsiella isolates were moderate catalase reactors. Escherichia and Shigella strains were mostly nonreactive, with less than one-third weekly (+) reactive, whereas most Enterobacter strains tended to be weakly reactive. Klebsiella strains were divided equally between nonreactive and weakly reactive. In practice, this test was also of great value in discerning nonpigmented Serratia cultured from the hospital environment and in detecting mixed flora containing nonspreading Proteus.     

Characterisation of a cysteine protease from bloodstream forms of Trypanosoma congolense.

A cysteine protease (trypanopain-Tc) with cathepsin-L-like properties has been purified from Trypanosoma congolense. The enzyme has an apparent molecular mass of 31-32 kDa by SDS/PAGE and 66 kDa by gel chromatography. It has a pI 7.4 and a high affinity for concanavalin A. Trypanopain-Tc catalyses the limited proteolysis of a variety of protein substrates such as fibrinogen, serum albumin and trypanosome variant-surface glycoprotein. It has minimal or no activity against casein or elastin. A variety of peptidyl amidomethylcoumarins and peptidyl diazomethanes were used to test the specificity of trypanopain-Tc. The better substrates had Arg or Lys in P1 and hydrophobic amino acids in P2 and P3. The best substrate found for trypanopain-Tc was Z-Phe-Arg-NHMec (Z, benzyloxycarbonyl; NHMec, 7-amido-4-methylcoumarin). The kinetic constants for the hydrolysis of Z-Phe-Arg-NHMec were kcat = 17.4 s-1, Km = 4.4 microM, kcat/Km = 4.0 microM-1.s-1, which are very similar to those of cathepsin L with this substrate. The specific substrates for cathepsin B (Z-Arg-Arg-NHMec) and cathepsin H (Arg-NHMec) were not hydrolysed by trypanopain-Tc under the conditions tested. The pH optimum of trypanopain-Tc against Z-Phe-Arg-NHMec was pH 6.0 but it showed a broad peak of activity extending well into the alkaline region. The enzyme was activated by low-molecular-mass thiol compounds and inhibited by cystatin, L-trans-epoxysuccinyl-4-guanidinobutane (E-64) and a variety of peptidyl diazomethanes. The most effective diazomethane inhibitors (Z-Leu-Leu-Met-CHN2, Z-Leu-Met-CHN2 and Z-Leu-Lys-CHN2, were inhibitory at nanomolar concentrations and were trypanocidal in vitro after 24-48 h incubation in greater than or equal to 20 microM [inhibitor]. However, it is not clear whether the trypanocidal activity of these inhibitors is a consequence of the inhibition of trypanopains or of some other essential proteolytic activities within the parasites.     

Sulfur chemistry,biofilm, and the (in)direct attack mechanism — a critical evaluation of bacterial leaching

It has been shown (a) that bacterial leaching of metal sulfides apparently requires the attachment of leach bacteria to metal sulfides, (b) that exopolymerbound iron compounds are responsible for or at least considerably increase the rate of the biological attack over the chemical rate, (c) that the primary attacking agent in leaching environments is the ferric iron hexahydrate ion, (c) that thiosulfate is the first intermediate sulfur compound, giving rise to a variety of other compounds including polythionate-containing periplasmic granula, and (d) that we have no idea about the actual concentrations of protons, ferrous/ferric and/or other cations, and sulfur compounds in the reaction space between the bacterium and the sulfide surface.     

MORPHOLOGICAL DIFFERENTIATION OF PROBOSCIDEA PARVIFLORA SSP. PARVIFLORA (MARTYNIACEAE) UNDER DOMESTICATION

Aboriginal groups in the southwestern United States have grown Proboscidea parviflora ssp. parviflora for basketry fiber. It has been hypothesized that cultivated plants of this subspecies with white seeds and long-clawed fruit (which provide the fiber) have been domesticated. Cluster, multiple discriminant, and unvariate statistical analyses presented here show that the putative domesticate is indeed morphologically distinct from “wild” plants. It has clearly been domesticated, and has diverged from wild populations to the extent that a new variety, var. Hohokamiana, is proposed for the domesticate. Wild and domesticated varieties differ especially in seed color, rostrum, crest, style, and anther lengths. Most characteristics of the wild variety seem to be genetically dominant to those of the domesticate, which may account for absence of a weedy form intermediate between the wild and domesticated varieties. The wild variety is morphologically more variable than the domesticate, probably because of its broader geographical range and its intergradation with another subspecies. Ethnological data and the variational patterns reported here suggest that the domesticate originated in southern Arizonanorthern Sonora, but probably not north of that region.     

Assembly Process Design for Managing Manufacturing Complexities Because of Product Varieties

This paper comprehensively studies assembly process design (APD) to handle variety and presents a new approach to strategically manage manufacturing complexities because of product varieties. It links the chronological steps of APD to a general approach with a view to reduce both short and long term wastes. It synthesizes two strategic approaches, product and process-based, while exploiting the APD of an entire product family. A new evaluation method for measuring the complexities of an assembly system at different stages of design has been introduced by applying information entropy. The developed evaluation methodology explains the available techniques related to design, interprets them from a design for variety point of view, and leads to techniques to manage complexities in manufacturing. The suggested methodology has been applied to examples from automobile part manufacturers and has been found to be effective in understanding the complexities in manufacturing and leading to some strategies to manage variety.     

The osmium tetroxide-potassium ferrocyanide (OsFeCN) staining technique for electron microscopy: A critical evaluation using ciliates,algae, mosses,and higher plants

Summary The osmium tetroxide-potassium ferrocyanide (OsFeCN)-method has been applied to a variety (24 objects) of ciliates, algae, mosses, and higher plants in order to test its specificity. The results showed big variation. Depending on the object, either general staining of the membranes, or selective staining of certain membranes, and/or staining of non-membraneous components in secretory vesicles and cell walls was obtained. In some cases, ultrastructural preservation was better than in controls. It is concluded that this staining technique may be helpful in tracing certain membrane systems, but it seems by no means specific for calcium-sequestering membrane systems.With support of the Deutsche Forschungsgemeinschaft     

Anesthetic inhibition of firefly luciferase, a protein model for general anesthesia, does not exhibit pressure reversal.

The surprising observation that pressures of the order of 150 atmospheres can restore consciousness to an anesthetized animal has long been central to theories of the molecular mechanisms underlying general anesthesia. We have constructed a high-pressure gas chamber to test for "pressure reversal" of the best available protein model of general anesthetic target sites: the pure enzyme firefly luciferase, which accounts extremely well for animal potencies (over a 100,000-fold range). We found no significant pressure reversal for a variety of anesthetics of differing size and polarity. It thus appears that either firefly luciferase is not an adequate model for general anesthetic target sites or that pressure and anesthetics act at different molecular sites in the central nervous system.     

EndoG is dispensable in embryogenesis and apoptosis

The mitochondrial protein, endonuclease G (EndoG), is one of the endonucleases implicated in DNA fragmentation during apoptosis. It has been shown to translocate from the mitochondria to the nucleus upon cell death stimuli. These observations suggest that EndoG is a mitochondrial cell death effector, and that it possibly acts as a cell death nuclease, similar to DNA fragmentation factor. To better understand the role of EndoG in development and apoptosis, we generated EndoG null mice by homologous gene targeting without disruption of D2Wsu81e. EndoG null mice are viable and develop to adulthood with no obvious abnormalities. Fibroblasts generated from the EndoG null mice show no difference in susceptibility when induced to die by a variety of intrinsic and extrinsic apoptotic stimuli. Additionally, EndoG null mice are equally sensitive to excitotoxic stress. These data suggest that EndoG is not essential for early embryogenesis and apoptosis.     

Interrelationships of selenium with other trace elements.

Biological interactions between selenium and a number of other elements occur that render selenium much less toxic than when it is present alone. These elements are arsenic, mercury, cadmium, and copper. Furthermore, the presence of selenium reduces the toxicity of mercury and cadmium. These are general biological interactions and have been found to occur in a number of animal species under a variety of conditions. It has been shown that the reaction products of selenium with mercury and cadmium are less toxic than an equal amount of selenium fed alone to chicks. The presence of arsenic shifts the excretion of selenium to the bile. There is no conclusive evidence that the presence of other elements reduces the absorption or retention of selenium. It is possible that some of the interactions are caused by the formation of a compound by selenium and other elements which has less affinity for active groups on biologically active compounds.--Hill, C.H. Interrelationships of selenium with other trace elements.     

Summary of recombinant human serum albumin development.

The methylotrophic yeast Pichia pastoris is well known as a host strain for the production of a variety of heterologous proteins. We have used P. pastoris for the production of recombinant human serum albumin (rHSA), for which we have developed efficient and specialized downstream processes. Results from structural analysis suggest that purified rHSA possesses an identical conformation to plasma derived human albumin (pdHA) and no difference from pdHA has been observed in neo-antigenicity. Host-cell-derived impurities (i.e. Pichia yeast component, DNA and mannan) have been evaluated in the purification process as well as in the drug substance and relevant specifications established. The efficacy and safety of rHSA have been tested in clinical studies and no difference from pdHA has been found in comparative study. Such studies have confirmed rHSA to have high efficacy with little or no adverse reaction.     

The Venetian Turpentine Mounting Medium

As a mounting medium to follow aceto-carmine the following modification of Zirkle's is suggested: Venetian turpentine, 25 ml.; phenol, 50 ml.; propionic acid, 35 ml.; acetic acid, 10 ml.; water, 20 ml. The technic can be employed with either root-tip or pollen-mother smears, and has been used with quite a variety of plants. It is especially valuable where it is desired to make temporary mounts permanent. The method is simple, and with reasonable care no displacement of marked cells occurs.     

Immunochemical analysis of CD107a (LAMP-1)

CD107a, also known as the lysosome associated membrane protein-1 (LAMP-1), is expressed largely in the endosome-lysosome membranes of cells, but is also found on the plasma membrane (1-2% of total LAMP-1). LAMP-1 has been implicated in a variety of cellular functions, including cancer metastasis. It has been proposed as a therapeutic agent for some cancers, and is a marker for lysosomal storage disorders and different cell types such as cytotoxic T cells. In light of this diversity of applications, it is important to have well characterized immune-reagents for the detection and quantification of LAMP-1. We have compared a new monoclonal antibody 80280 against LAMP-1 to an existing monoclonal antibody BB6 and a rabbit polyclonal antibody. While all antibodies gave similar results by immunofluorescence, the monoclonal antibody 80280 showed no epitope reactivity to LAMP-1 peptides, suggesting the possibility of a carbohydrate epitope. Western blotting revealed a weaker activity of the monoclonal antibody 80280 relative to either the BB6 monoclonal or the polyclonal antibodies. The monoclonal antibody 80280 is distinct from BB6, providing an additional reagent for CD107a analysis.     

Susceptibility to breast cancer and three polymorphisms of GSTZ1

Glutathione S-transferases class zeta (GSTζ) is involved in the detoxification of xenobiotic compounds and catalyzes the biotransformation of a variety of α-haloacids including dichloroacetic acid and chlorofluoroacetic acid. It has been reported that, in mice, deficiency of Gstz1 (a member of GSTζ) resulted in the generation of a constant level of oxidative stress. The present study was carried out to investigate the association between genetic polymorphisms of GSTZ1 (in promoter site G-1002A and in coding sites Glu32Lys and Gly42Arg) and risk of breast cancer. We included 106 females with breast cancer and 106 healthy females frequency matched for age. The study polymorphisms were not associated with risk of breast cancer (p>0.05). The polymorphisms of GSTZ1 showed strong linkage disequilibrium among cancer patients and control subjects (p<0.0001). There was no significant difference between cancer patients and controls for frequencies of the GSTZ1 haplotypes (p>0.05). It seems there is no meaningful relationship between the genetic polymorphisms of GSTZ1 and risk of breast cancer.     

Induction by ethylene of cyanide-resistant respiration

Ethylene and cyanide induce an increase in respiration in a variety of plant tissues, whereas ethylene has no effect on tissues whose respiration is strongly inhibited by cyanide. It is suggested that the existence of a cyanide-insensitive electron transport path is a prerequisite for stimulation of respiration by ethylene.     

Enzymatic Detection of the Growth of Staphylococcus aureus in Foods

A specific method has been developed for the extraction and measurement of staphylococcal nuclease in foods in which Staphylococcus aureus has grown. The method was used to compare staphylococcal growth with nuclease production in foods under varying conditions of temperature, aerobiosis, and competition from other microorganisms. It was concluded that the nuclease is produced under any conditions that permit growth of S. aureus, and little or no interference with the test was encountered either from mixed, natural populations or from a variety of pure, laboratory cultures. Nuclease and enterotoxin A production were shown to vary in synchrony for the 234 (Casman) strain of S. aureus, and the sensitivity of the enzymatic detection of nuclease was comparable to the sensitivity of serological detection of enterotoxin A. It was found that 15 min at 121 C was required to reduce the nuclease activity in slurries of contaminated ham below the level present in the unheated slurry. The extraordinary heat resistance of the nuclease permits its detection even in foods heated subsequent to the growth of S. aureus. The nuclease analysis requires about 3 hr to complete and requires no unusual equipment or reagents.     

绿色荧光蛋白及其应用

许多海洋无脊椎动物体内都含有绿色荧光蛋白,这种蛋白质结构很特殊,在受到激发时可以发射绿色或蓝色荧光。虽然对它的研究从本世纪六十年代才开始,但是它独特的性质逐渐引起了生物学界的广泛关注。本文将就绿色荧光蛋白的结构、性质及其应用前景作一综述。     

Determination of theophylline and its metabolites in rat liver microsomes and human urine by capillary electrophoresis

A capillary electrophoretic (CE) method has been developed for the determination of theophylline and all of its identified and potential metabolites. The method is rapid, resolves all metabolites to baseline, and requires extraction of only some biological fluids. It has been applied to the analysis of theophylline metabolism by hepatic microsomes from rats treated with a variety of inducing agents for different forms of P450 enzymes which metabolize theophylline, and to human urine spiked with theophylline and its metabolites, and concentrated by solid-phase extraction.     

Continuous recording of the formation of a photolabile fluorophore.

DNA immobilized on cellulose or agarose has been found very useful for the separation and purification of DNA-binding proteins (1–4).A variety of methods for DNA immobilization has been proposed, including absorption (3), entrapment in gels (4), and covalent binding (1).In the course of a study of the behavior of a complex between histones and immobilized DNA, we have met the need of a particularly stable DNA-cellulose preparation, i.e., involving not only covalent bonds between the nucleic acid and the polysaccharide, but bonds resistent to extremes of pH, particularly in the alkaline region.It has been found that cellulose activated with cyanuric chloride affords a convenient material for the preparation of DNA-cellulose, which has the following favorable characteristics: (i) It can be prepared by an easy and reproducible method; (ii) double-stranded DNA is bound in satisfactory amounts; (iii) the material is very stable, even in urea and in alkaline conditions.