Additive and synergistic interactions of monoclonal antibodies and immunotoxins reactive with breast and ovarian cancer.
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Animals↗
Biomedical subjects
Y Yu
Publications and source records attributed to Y Yu.
[Detection of specific DNA fragment of Plasmodium falciparum by solid phase PCR].
Solid phase PCR for detecting Plasmodium falciparum was established and used to detect cultured FCCl/HN isolate and malaria patients in Yunnan Province. The results revealed that the sensitivity of the method for detecting FCCl/HN isolate DNA was as minimal as 0.2 pg, or about 10 parasites. The specificity of the method was confirmed by discriminating malaria patients infected with different species of Plasmodium. The results suggest that solid phase PCR is a specific and sensitive method for early detection of P. falciparum.
Animals↗
Synthesis and cloning of the human lysozyme gene.
The human lysozyme is an enzyme with potential importance in clinical and industrial application. Owing to the limitation of its natural resource, we are making an attempt to produce the enzyme with the aid of recombinant DNA technology. Twenty-four segments with length ranging from 26 to 38 nucleotides were chemically synthesized by solid phase. The oligonucleotides were joined to form DNA duplexes by two different ligation methods. The entire gene covers a start signal ATG and a BamHI restriction site at its 5' end, and two stop signals TAA TGA and a SphI restriction site at its 3' end, besides the structural gene of human lysozyme. The synthetic gene was cloned into vector M13. The positive colonies were confirmed by dot-blot hybridization and analysis by restriction enzymes. The DNA sequence of the cloned enzyme gene was proved to be correct by M13 dideoxynucleotide chain termination method. The study on gene expression is under way.
Base Sequence↗
[Cloning and sequencing of a DNA fragment from Plasmodium falciparum FCC1/HN isolate].
A specific DNA fragment isolated from Plasmodium falciparum FCC1/HN isolate has been cloned in Bam H1 site of pUC18 and first partially sequenced by Sanger's method. The results show: G+C percent of DNA sequence is 26.8%, and the cloned DNA has many restriction sites which are conventional for subcloning. The authors suggest that this clone and sequence may be used as a guide for developing a DNA probe or PCR primers.
Animals↗
Yeast genes involved in growth inhibition by Pseudomonas syringae pv. syringae syringomycin family lipodepsipeptides.
Saccharomyces cerevisiae genes encoding functions necessary for inhibition by the Pseudomonas syringae pv. syringae cyclic lipodepsipeptide, syringomycin-E, were identified by mutant analyses. Syringomycin-E-resistant mutants were isolated, shown to contain single recessive mutations, and divided into eight gene complementation groups. Representative strains from five groups were resistant to nystatin, and deficient in the plasma membrane lipid, ergosterol. All of the mutant strains were resistant to the related cyclic lipodepsipeptides, syringotoxin and syringostatin. The findings show that: 1) at least eight gene-encoded functions participate in the inhibitory response to syringomycin; 2) ergosterol is important for this response; 3) the three related lipodepsipeptides have similar modes of action.
Anti-Bacterial Agents↗
Anisotropic rare-earth-Co exchange interactions in GdxY1-xCo5 and R2Co14B (R=Pr, Nd, or Gd).
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Journal Article↗
Drosophila evolution challenges postulated redundancy in the E(spl) gene complex.
The Enhancer of split [E(spl)] gene complex belongs to the class of neurogenic loci, which, in a concerted action, govern neurogenesis in Drosophila. Two genetically distinct functions, vital and neurogenic, reside within the complex defined by lethal mutations in the l(3) gro gene and by the typical neurogenic phenotype of deletions, respectively. Such deletions always affect several of the many embryonically active genes in the region, which cannot be mutated separately to lethality. Seven of these genes are extremely similar at the transcription and sequence level sharing the basic helix-loop-helix (bHLH) motif of transcriptional regulators. While these E(spl) bHLH genes seem to be required collectively for neurogenesis, they are nonessential individually, suggesting functional redundancy of the encoded gene products. No specific functions could yet be ascribed to any of the other genes located within the complex. One might expect these apparently dispensable genes, as well as the supposedly redundant bHLH genes, to be under little evolutionary constraint and, thus, to evolve most rapidly. However, we find the entire E(spl) gene complex highly conserved during Drosophila evolution, indicating that all the genes as well as their organization are of functional importance.
Amino Acid Sequence↗
Antibody-induced growth inhibition is mediated through immunochemically and functionally distinct epitopes on the extracellular domain of the c-erbB-2 (HER-2/neu) gene product p185.
Over-expression of the c-erbB-2 (HER-2/neu) gene product p185 occurs in 30% of breast and ovarian cancers. The p185 protein might serve as a target for serotherapy in that antibodies against different epitopes on the extracellular domain of p185 can inhibit growth of tumor cells in the absence of cellular or humoral effector mechanisms. To define epitopes of functional relevance, 11 monoclonal antibodies (MAbs) were evaluated for their ability to bind to the extracellular domain of p185. Results of competition studies with 125I-labeled and non-labeled antibodies indicated that 10 of 11 epitopes were grouped in a linear array. Antibodies against 7 epitopes inhibited anchorage-independent growth and antibodies against 2 of these epitopes also inhibited anchorage-dependent growth of SKBr3 breast-cancer cells that over-expressed p185. Treatment with antibodies exerted cytotoxic rather than cytostatic effects. When antibodies were used in combination, additive or supra-additive inhibition of anchorage-independent and anchorage-dependent growth was observed between pairs of antibodies. Growth inhibition did not relate to the affinity of the antibody or its isotype. Two antibodies that inhibited both anchorage-dependent and anchorage-independent growth also blocked binding of the HER-2/neu ligand, whereas 5 antibodies that inhibited only anchorage-independent growth had no effect on ligand binding. Inhibition of cell growth did not correlate with internalization of p185 or down-regulation of p185 on the cell surface. Fab fragments of active antibodies could also inhibit anchorage-independent growth of SKBr3. Thus, murine MAbs and their fragments recognized both immunochemically distinct and functionally distinct epitopes on the p185 molecule. Whereas inhibition of anchorage-dependent growth correlated with the ability of antibodies to block ligand binding, inhibition of anchorage-independent growth did not correlate with effects on ligand binding, internalization, cell-surface expression or cross-linking of p185.
Antibodies, Monoclonal↗
Retrospective cohort study of risk-factors for esophageal cancer in Linxian, People's Republic of China.
A retrospective cohort study of esophageal (including gastric cardia) cancer was conducted to examine dietary and other potential risk factors in Linxian, a high-risk area in P.R. China. Study subjects were identified based on participation in a cytology examination conducted in 1974. They were interviewed in 1989 to obtain information on esophageal cancer risk-factors and identify new cases and deaths. A total of 1,162 subjects from the analytic cohort of 12,693 were determined to have developed esophageal cancer over the 15-year follow-up period. Results indicate that increased age, male gender, a positive family history, low education level, surface-water use, and pork consumption were the strongest risk factors for esophageal cancer identified in this cohort, while use of corn as a primary staple and infrequent consumption of fresh vegetables also were possible risk factors. Traditional or suspected risk factors for esophageal cancer in this and other populations--smoking and alcohol use, and pickled vegetable and moldy food consumption--were not risk factors in this study. Some variation in risk was seen based on the subject's cytology result from 1974. We conclude that dietary factors appear to play a role in the etiology of esophageal cancer in this high-risk population, but are less important than other constitutional factors such as age, gender, and family history.
Adult↗
Analysis of host genetic control of scrapie-induced obesity.
The potential for induction of obesity during the preclinical phase of scrapie disease in mice was previously shown to be a function of both the strain of scrapie and the strain of inbred mouse. In the present study, host control of obesity induction by a scrapie strain was examined to determine if the effect were dependent on a single gene or multiple genes. The approach used was assessment of the pattern of weight induction in F1 and F2 crosses of parental inbred mouse strains that did or did not show a weight increase with a specific scrapie strain. Analyses of these data indicated that the induction of obesity was controlled by multiple host genes. In an unrelated observation, there was a correlation between the incubation period of a strain of scrapie in F2 generation mice and their coat color, i.e., the average incubation period of yellow-brown mice was significantly less than those of either black or white mice.
Animals↗
Dynamic observations on NK cell activity and IL-2 and IFN gamma production in vitro in patients on maintenance hemodialysis.
In order to study the immune function of patients on maintenance hemodialysis (MHD), we assayed NK cell cytotoxicity against K562 targets in 40 patients on MHD, and the production of IL-2 and IFN in peripheral blood mononuclear cells (PBMC) after PHA stimulation, in contrast to those in normal controls. The results showed that NK cell activity and IL-2 and IFN levels were markedly lower in the patients than in the controls. After a single dialysis, NK cell activity as well as IL-2 and IFN levels were elevated to different extent. But there was no significant change in patients after long-term dialysis. There was a positive correlation between the NK cell activity and IL-2 and IFN activity in the controls, but no such correlation was found in the patients on MHD. There was a positive correlation between the NK cell activity and IL-2 activity in patients after dialysis, suggesting that immune function were impaired in the patients on MHD, with a decline in the activity of NK cell and IL-2 and IFN, and a disorder of immune regulation cycle. These abnormal immune impairments in the patients could be partly corrected by hemodialysis. However, long-term hemodialysis is not much helpful in the improvement of patient's immune function.
Adult↗
Correlative factors of left ventricular hypertrophy in end-stage renal disease.
The correlative factors of left ventricular hypertrophy (LVH) in 287 end-stage renal disease (ESRD), admitted from 1983 to 1992, were analyzed. 52% of patients had LVH including 54% of patients on hemodialysis, 75% of patients with peritoneal dialysis and 38% of transplanted patients. Single factor analysis indicated that age, blood pressure, serum creatinine and BUN, hemoglobin, serum alkaline phosphatase, aortic valve disease and present DSRD therapy were related to LVH. Multiple logistic regression analysis indicated that the most important factors which independently related to LVH in all patients studied, were management of present ESRD treatment, age, hypertension and high serum alkaline phosphatase. In a subset of patients with severe LVH, high serum alkaline phosphatase level, high systolic blood pressure and age were the predictive factors. In patients on dialysis, the most important variable were age and high serum alkaline phosphatase. Hypertension was the sole predictor of LVH in the transplant group.
Adult↗
Multiplex DNA amplification and solid-phase direct sequencing for mutation analysis at the hprt locus in Chinese hamster cells.
We report here the development of multiplex in vitro DNA amplification and solid-phase direct exon sequencing for the analysis of mutations at the hypoxanthine-guanine phosphoribosyltransferase (hprt) locus in Chinese hamster cells. 18 representative HPRT-deficient mutants, derived either spontaneously, or after exposure to UV light or ionizing radiation, were analyzed. All 9 hprt exons were simultaneously amplified via the polymerase chain reaction (PCR) for rapid deletion detection. 5 mutants involve single- or multiple-exon deletions. Altered multiplex PCR patterns were detected in mutants Bsp-040, Bsp-065 and BGR-606. Subsequent direct sequence analysis reveals that Bsp-040 and Bsp-065 carry a 52-bp and a 13-bp intragenic DNA deletion in exon 3, respectively. BGR-606 contains a 223-bp insertion accompanied by a 10-bp deletion of intron sequence within exon 4 fragment. Other subtle DNA alterations identified by direct exon sequence analysis include single-base substitutions, small deletions and insertions, and RNA splicing mutations.
Animals↗
The effect of acidified glucose nutrient solution on preservation of platelet concentrate.
The preservation effect of a new platelet solution--acidified glucose nutrient solution (AGN)--was tested by adding AGN to pooled platelet rich plasma (PRP) prior to preparation of platelet concentrates (PC). Each PRP unit was prepared from a unit of whole blood and four PRP of the same blood group were pooled in 400 ml volume PVC bags. Equal aliquots of each pooled PRP were made prior to preparation of PC. AGN was added to one aliquot and nothing was added to the control aliquot. Equal volume and concentration PC were then prepared and the PC were further aliquoted for storage at 22 degrees C. After five days, the parameters of platelet count, pH, aggregability and hypotonic shock response (HSR) for PC preserved in AGN plasma were better than those of the controls preserved in normal ACD plasma. The aggregability and HSR of AGN platelet concentrates recovered to baseline after two h of incubation with fresh plasma. The results of electron microscopy show that platelets preserved in AGN have less changes in morphology. The results of our work suggest that the preservation effect of AGN on PC is similar to the effect of using second generation containers or preparation of platelets from thrombocytapheresis.
Blood Platelets↗
In vivo and in vitro studies of a copy number mutation of the RepA replication protein of plasmid pSC101.
The RepA replication protein of plasmid pSC101 binds as a monomer to three repeated sequences (RS1, RS2, and RS3) in the replication origin of the plasmid to initiate duplication and binds as a dimer to two inversely repeated sequences (IR1 and IR2) in its promoter region (D. Manen, L. C. Upegui-Gonzalez, and L. Caro, Proc. Natl. Acad. Sci. USA 89:8923-8927, 1992). The binding to IR2 autoregulates repA transcription (P. Linder, G. Churchward, G. X. Xia, Y. Y. Yu, and L. Caro, J. Mol. Biol. 181:383-393, 1985). A mutation in the protein RepA(cop) that affects a single amino acid increases the plasmid copy number fourfold. In vivo experiments show that, when provided in trans under a foreign promoter, the RepA(cop) protein increases the replication of a plasmid containing the origin of replication without repA, whereas it decreases the repression of its own promoter. In vitro experiments show that the purified RepA(cop) protein binds more efficiently to the repeated sequences within the origin than does RepA and that its binding to these sequences is more specific than that of RepA. Binding to an inversely repeated sequence within the repA promoter gives opposite results: the wild-type protein binds efficiently to that sequence, whereas the mutated protein binds less efficiently and less specifically. Footprint experiments confirmed these results and, in addition, showed a difference in the pattern of protection of the inversely repeated sequences by the mutant protein. Equilibrium binding experiments showed that the formation of protein-probe complexes at increasing concentrations of protein had a sigmoidal shape for binding to RS sequences and a hyperbolic shape for binding to IR sequences. The results, together with earlier work (G.-X. Xia, D. Manen, T. Goebel, P. Linder, G. Churchward, and L. Caro, Mol. Microbiol. 5:631-640, 1991), confirm that the binding of RepA to RS sequences plays a crucial role in the regulation of plasmid replication and that its binding to IR sequences plays a role in the autoregulation of RepA expression. They also demonstrate that the two separate functions of the protein are effected by two different forms of binding to the target sites.
Amino Acid Sequence↗
Multiple proteins interact with the fushi tarazu proximal enhancer.
The expression of the Drosophila segmentation gene fushi tarazu (ftz) is controlled at the level of transcription. The proximal enhancer, located approximately 3.4 kb upstream of the transcription start site, directs lacZ fusion gene expression in a ftz-like seven-stripe pattern in transgenic fly embryos. We have taken a biochemical approach to identify DNA-binding proteins that regulate ftz gene expression through the proximal enhancer. DNase I footprinting and methylation interference experiments with staged Drosophila embryo nuclear extracts identified nine protein binding sites in the proximal enhancer. Ten different sequence-specific DNA-binding complexes that interact with eight of these sites were identified. Some interact with multiple sites, while others bind to single sites in the enhancer. Two of the complexes that interact with multiple sites appear to contain the previously described ftz regulators, FTZ-F1 and TTK/FTZ-F2. These in vitro studies allowed us to narrow down the proximal enhancer to a 323-bp DNA fragment that contains all of the protein binding sites. Expression directed by this minimal enhancer element in seven ftz-like stripes in transgenic embryos is identical to that directed by the full-length enhancer. Internal deletions of several sites abolish reporter gene expression in vivo. Thus, the ftz proximal enhancer, like other cell-type-specific eukaryotic enhancers, interacts with an array of proteins that are expected to mediate the establishment, maintenance, and repression of transcription of the ftz gene in seven stripes in the developing embryo.
Animals↗
Quantitative and molecular analyses of genetic risk: a study with ionizing radiation.
Mammalian cells in culture have been used to study the genetic effects of physical and chemical agents. We have used Chinese hamster ovary (CHO) cells, clone K1-BH4, to quantify mutations at the X-linked, large (35 kb) hypoxanthine-guanine phosphoribosyltransferase (hprt) locus (the CHO/HPRT assay) induced by environmental agents. By transfecting an hprt-deletion mutant CHO cell line with the plasmid vector pSV2gpt, we isolated a transformant, AS52. AS52 cells carry a single functional copy of an autosomal, small (456 bp) xanthine-guanine phosphoribosyltransferase (gpt) gene (the bacterial equivalent of the mammalian hprt gene; AS52/GPT assay). We found that ionizing radiations such as X-rays and neutrons and oxidative genotoxic chemicals such as Adriamycin, bleomycin, hydrogen peroxide, and potassium superoxide are much more mutagenic to the gpt gene in AS52 cells than to the hprt locus in K1-BH4 cells. The hypermutability of the gpt gene probably results from a higher recovery of multilocus deletion mutants in AS52 cells than in K1-BH4 cells, rather than a higher yield of induced mutants. These results demonstrate that the use of the hprt locus alone could lead to an underestimate of the genetic risk of these agents. Analyses of the mutation spectrum using a polymerase chain reaction-based deletion screening and DNA sequencing procedure showed that a high proportion of HPRT- and GPT- mutants induced by X-rays carry deletion mutations. Thus, both the mutant frequency and mutation spectrum need to be considered in assessing the genetic risk of ionizing radiation and oxidative genotoxic chemicals.
Animals↗
Influence of amount and degradability of protein on production of milk and milk components by lactating Holstein cows.
Forty-one multiparous cows were utilized in a completely randomized design with a 2 x 2 factorial arrangement of treatments to evaluate the amount and degradability of dietary CP on production of milk and milk components. The TMR contained 25% alfalfa haylage, 25% corn silage, and 50% concentrate to provide either 16.4 or 19.4% CP with a calculated ruminal degradability of 55 or 70%. Intakes of DM, ADF, and NDF and BW were not different among treatments. Production of milk, 4% FCM, fat, CP, and SNF was not affected by amount or degradability of dietary CP. Milk CP percentage was not affected significantly by amount or degradability of CP. Milk fat percentage was increased by the diet that was high in ruminally undegradable protein (2.90 and 3.12; low and high ruminally undegradable protein, respectively). These data suggest that synthesis of milk and milk components was not limited by a shortage of AA or that the different dietary sources of supplemental CP did not alter AA availability.
Animal Feed↗