A full-length rabbit oviductin cDNA(1909bp) was cloned. It consists of a 5’-UTR of 52bp, an open reading frame (ORF) of 1374bp and a 3’-UTR of 483bp and has more than 80% homology with that of other mammal oviductin...A full-length rabbit oviductin cDNA(1909bp) was cloned. It consists of a 5’-UTR of 52bp, an open reading frame (ORF) of 1374bp and a 3’-UTR of 483bp and has more than 80% homology with that of other mammal oviductins. N-terminal peptide (NTP) (384 residues) and C-terminal peptide (CTP) (73 residues) of deduced protein precursor has about 80% and 50% identity with that of other mammals respectively. Fusion proteins GST-NTP 368(1R-368N)and GST-CTP73 (369F-441A) were expressed and purified. NH2-terminal of CTP sequencing reveals that the purified protein is consistent with the deduced one. In order to study the function of NTP and CTP the mouse anti-NTP and rabbit anti-CTP antisera were prepared. Tissue-specific (skeleton muscle, oviduct, uterus, ovary, liver, heart and brain) analysis indicated that rabbit oviductin was only found in oviduct. The conditioned medium derived from the rabbit oviduct mucosa epithelial cells has a function of overcoming the early embryonic development block of Kunming mous e cultured in vitro. Anti-CTP antiserum could totally inhibit the early embryo development at 2-cell stage cultured in the conditioned culture medium, but anti-NTP antiserum couldn’t. There was a positive relationship between the ratio of early embryos at development block and the dosage of anti-CTP antiserum added in the conditioned culture medium. These results suggest that oviductin has a function not only on fertilization, but also on the release of early embryonic development block, and the later function domain of rabbit oviductin may be situate in its C-terminal.展开更多
The objective in this experimental article is to gain evidential proof of near-dead cells, (sick-cells in relapse tumor) responding with recovery growth from special 4n, multi-chromatid chromosomes. Note, near-dead &l...The objective in this experimental article is to gain evidential proof of near-dead cells, (sick-cells in relapse tumor) responding with recovery growth from special 4n, multi-chromatid chromosomes. Note, near-dead </span><i><span style="font-family:Verdana;">normal human cells</span></i><span style="font-family:Verdana;"> with such converted chromosome structure gave rise to proliferative, fitness-gained, diploid </span><i><span style="font-family:Verdana;">first cells</span></i><span style="font-family:Verdana;">,</span><i> </i><span style="font-family:Verdana;">which</span><i> </i><span style="font-family:Verdana;">further gave rise to three different cell shape changed, recovery growth patterns. Previously, two cell shape changes had been recovered from same type normal human cells, transiently exposed to amino acid glutamine deficient growth medium with recovery growths also associated with presence of the special 4n cells. The 4n cell-division had been concluded to be a meiotic-like two-step division system to the fitness-gained diploid cells in numerous experiments. The main characteristi</span><span style="font-family:Verdana;">cs of this division system, was firstly whole genomes without polar oriented bent centromeres moving apart followed by much rarer simple fission division to two or three diploid cells, selectable for first cell proliferatio</span><span style="font-family:Verdana;">n. In general these 4n cells showed metaphase type rosette figures moving apart not in the normal spindle associated mitotic shape with centromeres polar-pointing with sloping arms. This sequence of events induced by glutamine-deficiency, was earlier shown to cause DNA breakage in metabolic studies however, the near-death condition was only assumed from normal fibro-blastic cell-sheet shrinkage. This was rectified by an RNA virus (Coxakie-B3), which virology known is a highly cell killing virus (4+ CPE on their scale). This virus replicates only in replicating cells, which led to recovery growths with progressive phenotypic cell-shape changes (spindle, polygonal and roundness cells), each intervened by “total” cell destruction. These three different growth patterns </span><span style="font-family:Verdana;">had morphologies, indistinguishably from today’s cancer diagnostic morphologies. “Mitotic” analyses of beginning growths for the three phenotypes revealed the special rosette figure separations from special 4n and higher ploidy level cells, and also total absence of spindle type mitoses. Tumorigenesis-relevant </span><span style="font-family:Verdana;">was centromere-puffing with premature chromatid separation, and chromatin compaction, a mechanism, that was suggested to protect the genome from damage (text). We suggest that the multi-chromatid polyploid cells with their genome reductive division system, can be a tractable </span><i><span style="font-family:Verdana;">in vitro</span></i><span style="font-family:Verdana;"> model system for therapy information, when repeated from a cell-killing agent, producing virus-free recovery growths. Will it be enacted upon? Not likely with profit-greedy industrial Goliath in the helm of cancer research. But, a not for profit cancer organization, could change this appalling situation.展开更多
Cave-adapted animals provide a unique opportunity to study the evolutionary mechanisms underlying phenotypic,metabolic,behavioral,and genetic evolution in response to cave environments.The Mexican tetra(Astyanax mexic...Cave-adapted animals provide a unique opportunity to study the evolutionary mechanisms underlying phenotypic,metabolic,behavioral,and genetic evolution in response to cave environments.The Mexican tetra(Astyanax mexicanus)is considered a unique model system as it shows both surface and cave-dwelling morphs.To date,at least 33 different cave populations have been identified,with phylogenetic studies suggesting an origin from at least two independent surface lineages,thereby providing a unique opportunity to study parallel evolution.In the present study,we carried out the most exhaustive phylogeographic study of A.mexicanus to date,including cave and surface localities,using two mitochondrial markers(cytochrome b(cyt b)and cytochrome c oxidase subunit I(COI))and nuclear rhodopsin visual pigment(rho).Additionally,we inferred the molecular evolution of rho within the two contrasting environments(cave and surface)and across three geographic regions(Sierra de El Abra,Sierra de Guatemala,and Micos).In total,267 individuals were sequenced for the two mitochondrial fragments and 268 individuals were sequenced for the rho visual pigment from 22 cave and 46 surface populations.Phylogeographic results based on the mitochondrial data supported the two-lineage hypothesis,except for the Pachón and Chica caves,whose introgression has been largely documented.The Sierra de El Abra region depicted the largest genetic diversity,followed by the Sierra de Guatemala region.Regarding the phylogeographic patterns of rho,we recovered exclusive haplogroups for the Sierra de El Abra(Haplogroup I)and Sierra de Guatemala regions(Haplogroup IV).Moreover,a 544 bp deletion in the rho gene was observed in the Escondido cave population from Sierra de Guatemala,reducing the protein from seven to three intramembrane domains.This change may produce a loss-of-function(LOF)but requires further investigation.Regarding nonsynonymous(dN)and synonymous(dS)substitution rates(omega valuesω),our results revealed the prevailing influence of purifying selection upon the rho pigment for both cave and surface populations(ω<1),but relaxation at the El Abra region.Notably,in contrast to the other two regions,we observed an increase in the number of dN mutations for Sierra de El Abra.However,given that a LOF was exclusively identified in the Sierra de Guatemala region,we cannot dismiss the possibility of a pleiotropic effect on the Rho protein.展开更多
基金Supported by National Natural Science Foundation of China (39730460)National "973" Project (G1999055902)National Labora-
文摘A full-length rabbit oviductin cDNA(1909bp) was cloned. It consists of a 5’-UTR of 52bp, an open reading frame (ORF) of 1374bp and a 3’-UTR of 483bp and has more than 80% homology with that of other mammal oviductins. N-terminal peptide (NTP) (384 residues) and C-terminal peptide (CTP) (73 residues) of deduced protein precursor has about 80% and 50% identity with that of other mammals respectively. Fusion proteins GST-NTP 368(1R-368N)and GST-CTP73 (369F-441A) were expressed and purified. NH2-terminal of CTP sequencing reveals that the purified protein is consistent with the deduced one. In order to study the function of NTP and CTP the mouse anti-NTP and rabbit anti-CTP antisera were prepared. Tissue-specific (skeleton muscle, oviduct, uterus, ovary, liver, heart and brain) analysis indicated that rabbit oviductin was only found in oviduct. The conditioned medium derived from the rabbit oviduct mucosa epithelial cells has a function of overcoming the early embryonic development block of Kunming mous e cultured in vitro. Anti-CTP antiserum could totally inhibit the early embryo development at 2-cell stage cultured in the conditioned culture medium, but anti-NTP antiserum couldn’t. There was a positive relationship between the ratio of early embryos at development block and the dosage of anti-CTP antiserum added in the conditioned culture medium. These results suggest that oviductin has a function not only on fertilization, but also on the release of early embryonic development block, and the later function domain of rabbit oviductin may be situate in its C-terminal.
文摘The objective in this experimental article is to gain evidential proof of near-dead cells, (sick-cells in relapse tumor) responding with recovery growth from special 4n, multi-chromatid chromosomes. Note, near-dead </span><i><span style="font-family:Verdana;">normal human cells</span></i><span style="font-family:Verdana;"> with such converted chromosome structure gave rise to proliferative, fitness-gained, diploid </span><i><span style="font-family:Verdana;">first cells</span></i><span style="font-family:Verdana;">,</span><i> </i><span style="font-family:Verdana;">which</span><i> </i><span style="font-family:Verdana;">further gave rise to three different cell shape changed, recovery growth patterns. Previously, two cell shape changes had been recovered from same type normal human cells, transiently exposed to amino acid glutamine deficient growth medium with recovery growths also associated with presence of the special 4n cells. The 4n cell-division had been concluded to be a meiotic-like two-step division system to the fitness-gained diploid cells in numerous experiments. The main characteristi</span><span style="font-family:Verdana;">cs of this division system, was firstly whole genomes without polar oriented bent centromeres moving apart followed by much rarer simple fission division to two or three diploid cells, selectable for first cell proliferatio</span><span style="font-family:Verdana;">n. In general these 4n cells showed metaphase type rosette figures moving apart not in the normal spindle associated mitotic shape with centromeres polar-pointing with sloping arms. This sequence of events induced by glutamine-deficiency, was earlier shown to cause DNA breakage in metabolic studies however, the near-death condition was only assumed from normal fibro-blastic cell-sheet shrinkage. This was rectified by an RNA virus (Coxakie-B3), which virology known is a highly cell killing virus (4+ CPE on their scale). This virus replicates only in replicating cells, which led to recovery growths with progressive phenotypic cell-shape changes (spindle, polygonal and roundness cells), each intervened by “total” cell destruction. These three different growth patterns </span><span style="font-family:Verdana;">had morphologies, indistinguishably from today’s cancer diagnostic morphologies. “Mitotic” analyses of beginning growths for the three phenotypes revealed the special rosette figure separations from special 4n and higher ploidy level cells, and also total absence of spindle type mitoses. Tumorigenesis-relevant </span><span style="font-family:Verdana;">was centromere-puffing with premature chromatid separation, and chromatin compaction, a mechanism, that was suggested to protect the genome from damage (text). We suggest that the multi-chromatid polyploid cells with their genome reductive division system, can be a tractable </span><i><span style="font-family:Verdana;">in vitro</span></i><span style="font-family:Verdana;"> model system for therapy information, when repeated from a cell-killing agent, producing virus-free recovery growths. Will it be enacted upon? Not likely with profit-greedy industrial Goliath in the helm of cancer research. But, a not for profit cancer organization, could change this appalling situation.
基金supported by the Project No.191986,Fronteras de la Ciencia-CONACyTPrograma de Apoyo a Proyectos de Investigación e Innovación Tecnológica(PAPIIT),UNAM No.IN212419。
文摘Cave-adapted animals provide a unique opportunity to study the evolutionary mechanisms underlying phenotypic,metabolic,behavioral,and genetic evolution in response to cave environments.The Mexican tetra(Astyanax mexicanus)is considered a unique model system as it shows both surface and cave-dwelling morphs.To date,at least 33 different cave populations have been identified,with phylogenetic studies suggesting an origin from at least two independent surface lineages,thereby providing a unique opportunity to study parallel evolution.In the present study,we carried out the most exhaustive phylogeographic study of A.mexicanus to date,including cave and surface localities,using two mitochondrial markers(cytochrome b(cyt b)and cytochrome c oxidase subunit I(COI))and nuclear rhodopsin visual pigment(rho).Additionally,we inferred the molecular evolution of rho within the two contrasting environments(cave and surface)and across three geographic regions(Sierra de El Abra,Sierra de Guatemala,and Micos).In total,267 individuals were sequenced for the two mitochondrial fragments and 268 individuals were sequenced for the rho visual pigment from 22 cave and 46 surface populations.Phylogeographic results based on the mitochondrial data supported the two-lineage hypothesis,except for the Pachón and Chica caves,whose introgression has been largely documented.The Sierra de El Abra region depicted the largest genetic diversity,followed by the Sierra de Guatemala region.Regarding the phylogeographic patterns of rho,we recovered exclusive haplogroups for the Sierra de El Abra(Haplogroup I)and Sierra de Guatemala regions(Haplogroup IV).Moreover,a 544 bp deletion in the rho gene was observed in the Escondido cave population from Sierra de Guatemala,reducing the protein from seven to three intramembrane domains.This change may produce a loss-of-function(LOF)but requires further investigation.Regarding nonsynonymous(dN)and synonymous(dS)substitution rates(omega valuesω),our results revealed the prevailing influence of purifying selection upon the rho pigment for both cave and surface populations(ω<1),but relaxation at the El Abra region.Notably,in contrast to the other two regions,we observed an increase in the number of dN mutations for Sierra de El Abra.However,given that a LOF was exclusively identified in the Sierra de Guatemala region,we cannot dismiss the possibility of a pleiotropic effect on the Rho protein.
