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61.
62.
High plasticity of multispecific DNA methyltransferases in the region carrying DNA target recognizing enzyme modules. 下载免费PDF全文
Multispecific cytosine C5 DNA methyltransferases (MTases) methylate more than one specific DNA target. This is due to the presence of several target recognizing domains (TRDs) in these enzymes. Such TRDs form part of a variable centre in the MTase primary sequence, which separates conserved enzyme core sequences responsible for general steps in the methylation reaction. By deleting, rearranging and exchanging several TRDs of multispecific MTases, we demonstrate their modular character; they mediate target recognition independent of a particular TRD or core sequence context. We show also that multispecific MTases can accommodate inert material of non-MTase origin within their variable region without losing their activity. The remarkable plasticity with respect to the material that can be integrated into this region suggests that the enzyme core sequences preceding or following it form separable functional domains. In spite of the documented flexibility multispecific MTases could not be endowed with novel specificities by integration of putative TRDs of monospecific MTases, pointing to differences between multi- and monospecific MTases in the way their core and TRD sequences interact. 相似文献
63.
Immunogold labelling of the cytoplasmic estradiol receptor in resting porcine endometrium 总被引:2,自引:0,他引:2
Summary Serial sections of resting porcine endometrium were analyzed with the monoclonal antibody 13H2 using goat antimouse IgG/5 nm gold as secondary reagent or with either polyclonal antibodies from goat #402 or the rat monoclonal antibody H222, both in combination with protein G/12 nm gold. A modestly higher labelling of nuclei than of cytoplasm was seen only with the monoclonal antibody H222. Polyclonal #402 and monoclonal 13H2 showed fewer attachments over nuclear than over cytoplasmic areas. The highest densities of attachment and of predominantly cytoplasmic labelling were obtained with the monoclonal antibody 13H2. The results confirm the earlier assumption of a restricted accessiblity of estradiol receptor in the cytoplasm of resting cells for immunoreagents. 相似文献
64.
Zulma Gazit David W. Weiss Daniel Shouval Michal Yechezkeli Volker Schirrmacher Michael Notter Jürgen Walter Eli Kedar 《Cancer immunology, immunotherapy : CII》1992,35(2):135-144
Summary The antitumor effects of chemotherapy, recombinant human interleukin-2 (IL-2), recombinant human interferon A/D (IFN), allogeneic human lymphokine-activated killer (LAK) cells, and antitumor monoclonal antibody (mAb), administered alone and in various combinations, were tested in athymic nude mice carrying human tumor xenografts. Treatment began 6–18 days after i.v. or i.p. inoculation of colorectal carcinoma or melanoma cell lines, when macroscopic growths were evident. Chemotherapy consisted of two or three courses of 5-fluorouracil (5-FU) or dacarbazine. IL-2 and/or IFN were administered three to five times weekly for 1–3 weeks, usually starting 2–5 days after chemotherapy. Human LAK cells were infused once or twice weekly for 2 or 3 weeks concurrently with IL-2. In some experiments, murine anticolorectal carcinoma mAb (SF25) was administered. In both tumor systems, chemotherapy alone or immunotherapy alone (IL-2, IL-2 + LAK cells, IFN, IL-2 + IFN ± LAK cells) had little or no therapeutic effects. Additive effects were obtained by combining chemotherapy with IL-2 and LAK cells or with IL-2 and IFN. In the majority of the experiments, the most effective combination was chemotherapy + IL-2 + IFN + LAK cells. Treatment with mAb was beneficial in the colorectal carcinoma system when combined with 5-FU + IL-2 or 5-FU + IL-2 + IFN. Homing experiments with radiolabeled human and mouse LAK cells injected i.v. showed increased early accumulation in the liver and lungs, whereas freshly explanted mouse splenocytes localized mostly in the spleen and liver. The tissue distribution pattern of human LAK cells was similar in normal and tumor-bearing mice (with lung metastases). These findings suggest that combination of chemotherapy with cytokines and LAK cells can be partially effective for advanced solid human tumors even in the absence of the host's T-cell immune response. Preliminary experiments showed that tumor-specific, anti-melanoma T-cell clones were effective in local (s.c.) tumor growth inhibition (Winn assay) following coinjection with the autologous tumor cells. 相似文献
65.
Regarding biological products, increasing awareness of potential side effects have placed great importance not only at protein
purity regarding other proteins but on the removal of biologicals such as DNA and especially virus the importance of which
may not be known. Monoclonal antibodies (Mab) have come to be an important class of molecules obtained from hybridoma cells,
i.e., nonrecombinant cells in culture. It has been noted during the last years, that with rare exceptions hybridoma cell lines
contain retrovirus like particles. The infectious nature of the EM-visible particles has been tested for, however, in most
cases not been substantiated. In order to bring these valuable biological reagents, Mab's, to good use in man for imaging
or therapy, the remaining concern about a potential retroviral infection has to be reduced to an acceptable minimum. We describe
experimental approaches for the validation of chromatographic and ultrafiltration steps used in the production of monoclonal
antibodies to remove and inactivate murine retrovirus.
Present day biotechnological manufacturing processes have been devised incorporating a number of strategic preventive measures
that have found wide spread acceptance. They permit to answer the question: how can a potentially harmful infection by an
unknown virus be excluded.
Knowledge of the efficacy of purification steps to clear infectious model virus is fundamental to devise biotechnological
manufacturing processes yielding a purified antibody for use in man. 相似文献
66.
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69.
Svenja Viete Walter Heiligenberg 《Journal of comparative physiology. A, Neuroethology, sensory, neural, and behavioral physiology》1991,169(1):15-23
Summary In its Jamming Avoidance Response (JAR), the gymnotiform electric fish Eigenmannia shifts its electric organ discharge (EOD) frequency away from similar interfering frequencies. Continual behavioral measurements were carried out in 164 juvenile fish until a correct JAR emerged. Sixty-four of these fish were raised in complete isolation, the remainder in a community of their siblings. A correct JAR emerged in fish of 1.2–1.6 cm in body length, corresponding to a developmental age of 24–32 days. In 6 of 164 fish, the emergence of a correct JAR followed an interim appearance of an incorrect JAR, which involved frequency shifts in the direction opposite to those of a correct JAR. The fish raised in isolation developed the same forms of behavior and showed the same sequence in their appearance as did socially raised fish. This indicates that the JAR and its developmental schedule are innate. The appearance of an incorrect JAR suggests initial errors or incompleteness in the wiring of central nervous connections. A correct JAR ultimately emerged even if a stimulus regimen was offered that rewarded frequency shifts in the direction opposite to those of a correct JAR. This indicates that the development of the JAR is immune to experimental alterations of sensory experience.Abbreviations
Df
frequency difference between a jamming stimulus and fish's EOD
-
ELL
electrosensory lateral line lobe
-
EO
electric organ
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EOD
electric organ discharge
-
JAR
Jamming Avoidance Response
-
nE
nucleus electrosensorius
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nE
subnucleus of nE, causing drop of EOD frequency
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nE
subnucleus of nE, causing rise of EOD frequency
-
Pn
pacemaker nucleus
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PPn
prepacemaker nucleus 相似文献
70.
Two population groups of Jammu and Kashmir (India) — Muslims and Bodhs-have been typed for haptoglobin and for CG and PI subtype
polymorphisms. The allele frequencies do not show significant differences between these two populations. HP and GC allele
frequencies of Bodhs and Muslims differ considerably from with that observed in other North Indian populations. The PI allele
frequencies of Bohds and Muslims differ considerably from those found in other Indian populations and are more similar to
Mongoloid ones. 相似文献