[Histological effect of SMANCS/LPD therapy in renal cell carcinoma].
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Adult↗
Biomedical subjects
H Maeda
Publications and source records attributed to H Maeda.
rec-A protein-promoted recombination reaction consists of two independent processes, homologous matching and processive unwinding. A study involving an anti-rec-A protein-monoclonal IgG.
recA protein promotes the formation and processing of joint molecules of homologous double-stranded DNA and single-stranded DNA. We studied the effects of an anti-recA protein monoclonal IgG (ARM193) on two processes carried out by the recA protein. The homologous matching, i.e. pairing of double-stranded DNA and single-stranded DNA by forming intermolecular base-pairing at homologous regions was found to occur even in the presence of an excess amount of antibody ARM193. On the other hand, processive unwinding, i.e. the propagation of the unwinding of double-stranded DNA through a processive reaction of recA protein, which occurs even in the absence of single-stranded DNA, was found to be very sensitive to the inhibition by antibody ARM193. Therefore, we conclude that homologous matching and processive unwinding are independent of each other. Analysis of the effect of antibody ARM193 on the various activities of recA protein suggests that the entire reaction of the formation of joint molecules and their processing can be rationalized in terms of these two underlying processes, homologous matching and processive unwinding. This analysis also suggests that homologous matching seems to require only the binding itself of active units of recA protein to single-stranded DNA but not necessarily either the cooperativity of the protein or unwinding.
Animals↗
Enzymatic assay of hemoglobin in tissue homogenates with chlorpromazine.
An assay of hemoglobin in tissue homogenates is described. Tissues are homogenized with a 40 mM phosphate buffer (pH 8.0) solution without the use of a polytron or sonicator. The assay should be carried out before fractionations such as centrifugation. By measuring the free-radical intermediates of chlorpromazine, which are formed by the addition of hemoglobin, a linear calibration curve can be obtained within the range of hemoglobin contents between 0.8 and 12.0 micrograms. The present method can be applied to the assay of hemoglobin in several kinds of tissues from rats.
Animals↗
Internalization of serratial protease into cells as an enzyme-inhibitor complex with alpha 2-macroglobulin and regeneration of protease activity and cytotoxicity.
Extracellular serratial protease (56,000 Da) is known to be cytotoxic. Fluorescein isothiocyanate-labeled protease was found to form a complex with human alpha 2-macroglobulin (alpha 2M), and this enzyme-inhibitor complex was purified. The protease was found to be internalized by fibroblasts in culture as a complex with alpha 2M, which resulted in cell destruction. Regeneration of enzyme activity was confirmed in cells after 2-3 h of incubation. Chicken egg-white ovomacroglobulin, a homolog of human alpha 2M, formed a complex with this enzyme similarly and more tightly but failed to exhibit protease activity, cytotoxicity, and internalization into cells.
Binding, Competitive↗
Binding to and internalization by cultured cells of neocarzinostatin and enhancement of its actions by conjugation with lipophilic styrene-maleic acid copolymer.
The binding of a copoly(styrene-maleic acid)-conjugated neocarzinostatin (NCS) designated as smancs (Mr 16,000), and parental NCS (Mr 12,000) to cultured cells was investigated. These drugs were labeled with fluorescein isothiocyanate which retained biological activity and were used for binding studies. The binding of these drugs to HeLa cells was dependent on time and temperature, with 2 times more drug being bound at 37 degrees C than at 0 degree C. In the presence of a 100-fold molar excess of unlabeled NCS, the binding of smancs or NCS to HeLa cells was inhibited similarly. Therefore, it was suggested that smancs binds to NCS-binding sites (receptor) of the cell surface. However, the amount of cell-bound smancs was increased about 20-fold compared with that of NCS. Scatchard plot analyses of the binding of these drugs to HeLa and WISH cells indicated that this increase was due to alterations in affinity resulting from polymer conjugation of the drug to receptor rather than to an increase in the number of drug-binding sites at the cell surface. Furthermore, when the cytotoxicity of these drugs to HeLa cells was compared, smancs needed only 5 min to achieve 50% inhibition of the control. In contrast, the same dose level of NCS required more than 90 min to achieve the same toxic effect. More rapid internalization of smancs than NCS was also elucidated under fluorescence microscopy at 37 degrees C. There was no intracellular incorporation of these drugs below 20 degrees C. These results indicated that an increased lipophilicity of smancs appears to be responsible for its increased cell surface affinity, internalization rate, and toxic effect. Concordant to this interpretation was that among various copoly(styrene-maleic acid) esters, a more hydrophobic derivative showed more internalization (butyl greater than ethyl greater than carboxylate). These results suggest the possibility of utilizing protein tailoring to augment the subcellular activity of functional proteins.
Antibiotics, Antineoplastic↗
Anticancer effects of arterial administration of the anticancer agent SMANCS with lipiodol on metastatic lymph nodes.
A new method of arterially administering an oily anticancer agent was successfully established for the selective targeting of metastatic lymph nodes. A high molecular weight anticancer agent, a conjugate of copolymer (styrene maleic acid) to neocarzinostatin (SMANCS) was prepared in our laboratory and dissolved in a lymphographic oily contrast medium, Lipiodol (SMANCS/Lipiodol). SMANCS/Lipiodol was administered intraoperatively to eight patients with colorectal cancer and preoperatively to one patient with gastric cancer with lymph node metastases. In six of the patients with colorectal cancer, the drug was administered via an artery and in the other two patients the drug was injected into the wall of the colon near the primary cancer. In the patient with gastric cancer, the drug was administered via the left gastric artery. Delivery of the drug to the lymph nodes was examined roentgenologically and the anticancer effect was examined histologically. The results showed that SMANCS/Lipiodol could be delivered to the metastatic lymph node via the artery, but it could not be delivered to the metastatic lesion of the lymph node via the lymphatic route. In the patient with gastric cancer, SMANCS/Lipiodol preoperatively administered via an artery was found to remain selectively in a metastatic lymph node and an anticancer effect was histologically proved in all three of the metastatic lymph nodes.
Antibiotics, Antineoplastic↗
Nuclear factors binding to the human immunoglobulin heavy-chain gene enhancer.
The human immunoglobulin heavy chain (IgH) gene contains at least two tissue-specific regulatory regions, which are similar to the mouse IgH gene. One is the J-C enhancer and another is located in the 5' promoter region. Using an electrophoretic mobility shift assay and DNase I footprint, we have examined the interaction of factors in B cell nuclear extracts with the two regulatory regions of the human IgH gene. We have identified a nuclear factor in mouse B cell nuclear extracts which bound to specific sequence in the human IgH enhancer. This factor is apparently not present in mouse fibroblast nuclear extracts. We also found factor(s) which bound to the highly conserved octanucleotide sequence within the human IgH enhancer and 5' promoter regions.
Animals↗
Dopamine agonists produce functional recovery from septal lesions which affect hypothalamic defensive attack in cats.
Effects of lesions of the lateral septum and subsequent administration of methamphetamine (MAT, 1 mg/kg, i.p.) or apomorphine (APO, 1 mg/kg, i.p.) on thresholds for defensive attack elicited by electrical stimulation of the ventromedial hypothalamic nucleus (VM) were examined. Hissing and directed attack were selected for threshold determination. Thresholds were measured under two situations: one with provocation by a human and the other without it. Electrolytic lesions of the lateral septum enhanced the facilitative influences exerted by the provocation on the thresholds, however, subsequent administration of MAT or APO abolished or tended to abolish the enhancement. The rapid recovery of function was interpreted to have taken place due to excessive dopaminergic inputs to the spared tissue of the lateral septum, and a gating mechanism of neuronal information by dopamine was suggested.
Aggression↗
Reduction of hepatic metastases in rabbits by administration of an oily anticancer agent into the portal vein.
We studied a prophylactic chemotherapy against hepatic metastases arising from the shedding of tumor cells into the portal circulation. The therapy was done with a lymphographic oily contrast medium, Lipiodol, and a high molecular weight anticancer agent named poly(styrene-maleic acid) copolymer conjugated neocarzinostatin (SMANCS), developed in our laboratory. SMANCS was dissolved in Lipiodol by sonication (SMANCS/Lipiodol, 1 mg of SMANCS in 1 ml of Lipiodol). Twelve rabbits were simply inoculated with the highly malignant carcinoma VX-2. Fifteen rabbits were given injections of SMANCS in glucose and Lipiodol into the portal vein and were subsequently inoculated with the tumor cells. Eighteen were given injections of SMANCS/Lipiodol and then the tumor cells. These rabbits were killed 12 days later. Thirteen were given injections of the tumor cells alone and were allowed to survive. Sixteen were given injections of SMANCS/Lipiodol and then with the tumor cells; they were allowed to survive. Rabbits given injections of SMANCS/Lipiodol before tumor inoculation had significantly fewer (P less than 0.001) metastases than those not treated or those given SMANCS in glucose and Lipiodol. Survival was significantly longer [P less than 0.005; 36.0 +/- 7.7 (SD) days] with SMANCS/Lipiodol before tumor inoculation than without treatment [23.5 +/- 3.0 days]. SMANCS/Lipiodol has a prolonged anticancer effect because it remains in the portal vein and allows sustained drug release from the oil (Lipiodol) to aqueous spaces. Hepatic metastases might be prevented by portal administration of the appropriate oily anticancer agent.
Animals↗
Antitumor activity of some bacterial proteases: eradication of solid tumors in mice by intratumor injection.
A protease isolated from the culture filtrate of a Gram-negative bacteria, Serratia marcescens kums 3958, showed very potent antitumor activity when injected into Meth-A or RL male 1 tumors in BALB/c mice at 30 micrograms per tumor or more. This and certain other proteases, which are resistant to many protease inhibitors in plasma, appear to be new candidate drugs for regional treatment of solid tumors.
Animals↗
Regulation of immunoglobulin gene transcription by labile repressor factor(s).
The cloned human gamma 1 heavy chain gene (HIG1) was scarcely expressed in the stable transformants of a mouse fibroblast line, L cell or a T cell line, EL4, and no gamma 1 heavy chain was produced in these cells. Upon treatment with cycloheximide or other kinds of protein synthesis inhibitors, the transcription of HIG1 gene was induced in L cell transformants as well as in T cell transformants. Transcription rate of bacterial gpt gene, which was derived from the plasmid vector used for transfection of HIG1 gene and located just upstream of HIG1 in the transformants, was also greatly enhanced after cycloheximide treatment. But the expression of several endogenous genes in the L cells tested was not affected by the cycloheximide treatment. Nuclear transcription assay indicated that the appearance of HIG1 gene transcripts after treatment with cycloheximide was mainly due to the induction of the transcription. Deletion of an enhancer element from HIG1 gene lowered the inducing activity of cycloheximide in the L cell transformants, but a low level of HIG1 gene expression was still observed. These results suggested that trans-acting factors similar to those present in B lymphoid cells are also functioning in non-B lymphoid cells but their activity is inhibited by short-lived repressor protein(s). Such repressor protein(s) appear to act on regulatory element(s) of the immunoglobulin heavy chain gene including enhancer region as well as 5' flanking region.
Animals↗
The cis-acting regulatory elements of immunoglobulin heavy chain gene involved in enhanced immunoglobulin production after lipopolysaccharide (LPS) stimulation.
In the present study, the transient expression of the human immunoglobulin heavy chain gene (HIG1) was analyzed in a mouse pre-B-cell line, 70Z/3, after LPS stimulation. HIG1 gene and its recombinant plasmids were transfected by the calcium phosphate method into 70Z/3 cells and the cells were stimulated with lipopolysaccharide (LPS) 48 hr after DNA transfection. The amounts of human heavy chain gene products greatly increased in 70Z/3 cells after LPS stimulation, but the increment was diminished by deletion of the heavy chain gene enhancer element (MluI-HpaI fragment) in the J-C intron from the HIG1 gene. The gene, delta 3, which contained the 5' promoter region and the rearranged VDJ region of HIG1 but lacked the enhancer element, was weakly transcribed in 70Z/3 cells after LPS stimulation. Insertion of the enhancer element into the delta 3 gene greatly enhanced the transcription of the VDJ gene. The highest enhancement of the VH gene transcription rate was obtained when the 3' half of the enhancer element was ligated to the delta 3 gene. The present data suggest that the 3' half of the enhancer element of the heavy chain gene may play an important role in the enhanced production of immunoglobulin which is induced with LPS stimulation.
Animals↗
Coimmobilized system of NAD with dehydrogenases.
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Alcohol Dehydrogenase↗
Immunohistochemical studies of major histocompatibility antigens in a case of the bare lymphocyte syndrome without immunodeficiency.
A case of the bare lymphocyte syndrome without apparent immunodeficiency was observed. The patient had, in addition, ichthyosis vulgaris and sinobronchial disease. A pustular lesion developed at first on the anterior aspect of lower part of the left leg. This lesion gradually increased in size and evolved into giant indurated, irregular adjoining plaques. On biopsy, the histologic findings were similar to necrobiosis lipoidica. No human lymphocyte antigen (HLA) class I antigens were detected on peripheral mononuclear cells; however, HLA-DR antigens were present on B lymphocytes. Immunohistochemical studies disclosed defective expression of class I antigens in the non-lesional skin, but positive expression was demonstrated in the lesional area. HLA-DR antigens were expressed on keratinocytes and on most infiltrating inflammatory cells in the affected skin. It is therefore speculated that class I antigen appearance and mononuclear cell infiltrate each induces the other and that together they play an important role in the formation and enlargement of the skin lesion.
Adult↗
Different binding kinetics of Serratia 56K protease with plasma alpha 2-macroglobulin and chicken egg white ovomacroglobulin.
We recently reported that Serratia 56K protease is inhibited by plasma alpha 2 macroglobulin (alpha 2M) temporarily and by chicken egg white ovomacroglobulin (ovoM) continuously (Molla, A. et al. (1986) Infect. Immun. 53, 522-529). The inhibition of this protease is almost complete with ovoM whereas it is incomplete with alpha 2M, although these two macroglobulins show homology and many similarities. In the present study we determined the apparent numbers of binding sites and binding constants for the two macroglobulins by means of the fluorescence polarization method using FITC-labeled 56K protease. The time courses of complex formation of 56K protease with alpha 2M and ovoM were different; with ovoM it was complete within 5 min while with alpha 2M 150 min was required. Their apparent molecular volumes were also different; the fluorescence polarization value of the E/I complex was 18.7% larger with ovoM than with alpha 2M. The association constants obtained on Scatchard plot analysis with 56K protease and alpha 2M or ovoM were 0.33 X 10(7) M-1 and 1.09 X 10(7) M-1, respectively. One molecule of each of these macroglobulins binds 1.13 and 1.35 molecules of 56K protease, respectively. Upon E/I complex formation, an increase in amino groups due to proteolysis was noted in both cases, but more progressive proteolysis was observed in the case of alpha 2M. Furthermore, when the 56K protease was inactivated through the depletion of Zn atoms, complex formation did not occur.
Amino Acids↗
Malignant schwannoma of the maxilla in a patient without neurofibromatosis.
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Diagnosis, Differential↗
Effects of CuCl2 on the dimer-tetramer equilibrium of concanavalin A.
Tetrameric concanavalin A at neutral pH dissociated into the dimer when CuCl2 was added in a concentration range comparable to protomer concentration. This effect of CuCl2 was largely suppressed when NaCl concentration was increased. Neither the conformation of the protein nor its binding activity to 4-methylumbelliferyl alpha-D-mannopyranoside was affected on addition of CuCl2.
Concanavalin A↗
Nonimmunologic hydrops fetalis and chromosome aberration.
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Amniocentesis↗