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At least 937 records · Page 52Linked to original sources
PubMed1998

Inhibition of influenza virus RNA polymerase by 5'-capped short RNA fragments.

We have demonstrated that 5'-capped short RNA fragments inhibit the expression of chloramphenicol acetyltransferase (CAT) in the murine 76 cell line, derived which expresses the genes for the RNA polymerases (PB1, PB2, and PA) and the nucleoprotein (NP) of influenza virus in response to treatment with dexamethasone. We have synthesized 5'-capped short RNA fragments (8-13 ntds long) with a 5'-capped structure (m7GpppGm) using T7 RNA polymerase. The 5'-capped short RNA fragments (8-13 ntds long) were encapsulated in liposomes and were tested for their inhibitory effect by a CAT-ELISA assay using the clone 76 cells. The RNA fragments that were 9-12 ntds long showed inhibitory effects. In particular, the 9 ntds long RNA fragment, was highly inhibitory. On the other hand, the inhibitory effect of the 13 ntds long RNA fragment was considerably decreased in comparison with the other short RNA fragments. The minimal RNA chain length required for priming activity was found to be 12 ntds long. Furthermore, the 5'-capped RNA fragments exhibited higher inhibitory activities than the antisense phosphorothioate oligonucleotide (PB2-AUG-as, 20 ntds long) complementary to the site of the PB2-AUG initiation codon. Liposome encapsulation protected the RNA fragments in serum-containing medium and substantially improved their cellular accumulation.

DNA-Directed RNA Polymerases↗
PubMed1996

Two-dimensional separation and cloning of chromosome 1 NotI-EcoRV-derived genomic fragments.

The two-dimensional (2-D) separation of genomic digests has provided the means to analyze over 2000 unique restriction fragments simultaneously in a single gel, for genetic variation as well as for genomic alterations in cancer. By utilizing different combinations of restriction enzymes or different electrophoretic conditions, the number of analyzable fragments in multiple 2-D patterns can be augmented. We have previously shown the feasibility of distinguishing between spot intensities representing fragments from one allele and from two alleles and have implemented approaches for the cloning of fragments of interest in 2-D gels. In this study, the 2-D separation and cloning of chromosome 1 NotI-EcoRV-derived genomic fragments was performed. Three hundred forty-six NotI fragments in whole genomic preparations were assigned to chromosome 1. To verify the reliability of the assignment, two of the NotI fragments attributed to chromosome 1 were cloned and sequenced. The fragments that contained CpG islands were mapped by FISH to 1p35-p36.1 and to 1p13.3-p21, respectively. Our study indicates the feasibility of analyzing 2-D separations of whole genomic digests for the detection of alterations in specific chromosomes. The large number of restriction fragments attributed to chromosome 1 provides the means to screen 2-D patterns for chromosome 1 deletions and amplifications with a high marker density.

Cell Line↗
PubMed1978

Periodicity and fragment size of DNA from mouse TLT hepatoma chromatin and chromatin fractions using endogenous and exogenous nucleases.

The action of micrococcal nuclease, DNase I and DNase II on mouse TLT hepatoma chromatin revealing the periodicity of its structure as visualized by denaturing and non-denaturing gel electrophoresis, was consistent with the action of these enzymes on other chromatins. Micrococcal nuclease showed a complex subnucleosome fragment pattern based on multiples of 10 base pairs with a prominant couplet at 140/160 base pairs and the absence of the 80 base pair fragment. This couplet of the core and minimal nucleosome fragments was conspicuously present in the mononucleosomes found in the 11S fractions of a glycerol gradient centrifugation. DNase I and II produced a fairly even distribution of a 10 base pair increasing series of fragments to about 180 base pairs, a pattern also repeated in the DNA of nucleosome glycerol-gradient fractions. In limited digestions by these nucleases multinucleosomic DNA fragments are pronounced. These fragment lengths are multiples of an estimated average repeat length of nucleosome DNA of 180 base pairs. The action of the endogenous Mg/Ca-stimulated endonuclease produced only limited cuts in the hepatoma chromatin resulting primarily in multi-nucleosomic DNA fragment lengths and only upon lengthy digestion limited subnucleosomic, 10-base-pair multiple fragments are produced. The putative euchromatin-enriched fractions (50-75S) of the glycerol gradient centrifugation of autodigested chromatin, similarly, contained primarily the multinucleosomic DNA fragment lengths. These results are consistent with our previous electron microscopic demonstration that autodigested chromatin as well as the putative euchromatin-enriched fractions were composed of multi-nucleosomic chromatin segments containing a full complement of histones.

Animals↗
PubMed1991

Stability of osteochondral fragments of the femoral condyle: magnetic resonance imaging with histopathologic correlation in an animal model.

The stability of surgically induced osteochondral fragments of the femoral condyle was examined by magnetic resonance imaging (MRI) using T1- and T2-weighted spin echo sequences in 7 dogs; contrast-enhanced T1-weighted spin-echo sequences were also obtained. Animals were sacrificed between the 34th and 196th day after surgery. MR images were compared with the histopathologic findings. Two loose and five stable fragments were found after injection of contrast medium. With the loose fragments, a well-defined line of high signal intensity between the fragment and the epiphysis showed marked enhancement. Histological examination revealed vascularized granulation tissue at the interface. Stable fragments also showed a similar, but irregularly defined line on plain sequences, but no enhancement after injection of contrast medium; histological examination showed no granulation tissue at the interface but intact bone trabeculae within the completely repaired fracture. Fibrocartilaginous repair at the articular cartilage surface also showed enhancement. Contrast-enhanced MR imaging allowed an exact delineation of the line of separation of unstable osteochondral fragments in this animal model with differentiation from a similar line occurring in stable fragments. However, this interface line in relation to stable fragments could not be explained histologically and probably reflects differences of binding or distribution of protons in healing osteochondral fragments.

Animals↗
PubMed2004

Social fragmentation, severe mental illness and suicide.

BACKGROUND: Geographic patterns of suicide are associated with area levels of social fragmentation. It is unknown whether this reflects higher levels of severe mental illness in socially fragmented areas. METHOD: Data on psychiatric inpatient admissions and suicides amongst people aged 15-64 living in the City of Bristol [1991-1992] were postcode matched to the city's 34 electoral wards. Ecological associations of psychiatric admission (used as a 'proxy' measure of prevalence of severe mental illness) and suicide rates with levels of social fragmentation were investigated using negative binomial regression models. RESULTS: Psychiatric hospital admission rates were higher in areas with high levels of socioeconomic deprivation than in areas with high levels of social fragmentation. In contrast, associations with suicide were stronger in relation to social fragmentation than socioeconomic deprivation. Association of suicide with social fragmentation was only moderately attenuated in models controlling for psychiatric admission rate and socio-economic deprivation, RR 1.23 (95 % C. I. 1.09-1.38) per quartile increase in social fragmentation, compared to 1.29 (95% C. I. 1.16-1.44) before adjustment. CONCLUSION: The association between social fragmentation and suicide is not explained by socioeconomic deprivation or the prevalence of severe mental illness within socially fragmented areas as measured by psychiatric admission rate.

Adolescent↗
PubMed2001

Traditional land-use systems and patterns of forest fragmentation in the highlands of Chiapas, Mexico.

The influence of slash-and-burn agriculture and tree extraction on the spatial and temporal pattern of forest fragmentation in two municipalities in the highlands of Chiapas, Mexico was analyzed. The data series were derived from two subsets of satellite images taken in 1974 and 1996. The analysis was based on area, edge, shape, core area, and neighbor indices. During the 22 years, the dense forest decreased by 8.9%/yr in Huistan and by 8.6%/yr in Chanal, while open/disturbed forest, secondary vegetation, and developed area increased in both municipalities. The total number of fragments increased by 1.4%/yr and 2.3%/yr in Huistan and Chanal, respectively. Dense forest showed the highest increase in the number of fragments (6%/yr in Huistan and 12%/yr in Chanal), while edge length, core area, and number of dense forest core areas decreased. The larger fragments of dense forest present in 1974 were divided into smaller fragments in 1996; at the same time, they experienced a process of degradation toward open/disturbed forest and secondary vegetation. Two different fragmentation patterns could be distinguished based on agricultural or forestry activities. Forest fragmentation did not occur as a continuous process; the pattern and degree of fragmentation were functions of land tenure, environmental conditions, and productive activities. The prevalence of rather poor soil conditions, small-holdings, growing human population densities, increasing poverty, and the absence of alternative economic options will maintain a high rate of deforestation and forest fragmentation in the studied region.

Agriculture↗
PubMed1998

The fragmented neuronal Golgi apparatus in amyotrophic lateral sclerosis includes the trans-Golgi-network: functional implications.

The Golgi apparatus (GA) of spinal cord motor neurons is fragmented in sporadic amyotrophic lateral sclerosis (ALS), and in asymptomatic and symptomatic transgenic mice expressing the G93A mutation of the gene of the human Cu,Zn superoxide dismutase, found in certain cases of familial ALS (FALS) [Gonatas NK (1994) Am J Pathol 145:751-761; Mourelatos Z, et al. (1996) Proc Natl Acad Sci USA 93:5472-5477]. A similar fragmentation of the GA has been described in cells treated with microtubule-depolymerizing drugs, where the organelle is functional and contains both Golgi stacks and trans-Golgi network (TGN), the compartment of exit and targeting of proteins processed by the GA. To gain a better definition of the structure of the fragmented neuronal GA in ALS, four cases of sporadic ALS with numerous Bunina bodies in spinal cord motor neurons were stained with antibodies against human TGN and against the lumenal and cytoplasmic domains of MG160, a protein of the medial cisternae of the GA. The fragmented GA was stained with the three antibodies, indicating the presence of both Golgi stacks and TGN. Furthermore, the staining of the fragmented GA by the antiserum against the cytoplasmic domain of MG160 indicates that the fragmentation of the GA is not the result of a terminal and global cytoplasmic lytic event. The Bunina bodies were not stained by the anti-MG160 antibodies, suggesting that they are not derived from the GA. The perikarya of neurons with fragmented GA showed normal immunoreactivity with antibodies against the heavy neurofilament subunit and alpha-tubulin. However, because of the lack of appropriate antibodies the localization of proteins such as spectrin, ankyrin, centractin and others which link the microtubules with the GA were not done. The findings support the hypothesis that, in ALS, the fragmented neuronal GA is functional. Additional work with animal models of ALS may establish whether the fragmentation of the GA is a sign of early degeneration or a compensatory reaction of the injured motor neuron.

Aged↗
PubMed2002

What is pathological with gaze shift fragmentation in Parkinson's disease?

Oculomotor dysfunction in Parkinson's disease (PD) is mainly characterized by a fragmentation of memory-guided gaze shifts (target is reached by several hypometric saccades). Since this phenomenon can also be observed in normal subjects, we scrutinized its pathophysiological significance in PD patients. We recorded horizontal eye movements in eleven mildly- or moderately-affected PD patients and eleven control subjects. A quantitative assessment of gaze shift fragmentation was made possible by increasing its incidence over a sequence of two visually- and two subsequent memory-guided gaze shifts. Basic saccade measures (latency, velocity, etc.) were similar in the two subject groups as well as in fragmented versus non-fragmented gaze shifts. Fragmentation probability is increased in the second memory-guided gaze shift, and this clearly more so in patients than in controls. The fragmentation shows a typical gain pattern (uniform increase of gain of saccadic amplitudes across correction saccades towards 1.0 with the last saccade of the gaze shift) independent of subject group, stimulus mode, and fragmentation degree. Gaze shift fragmentation represents a physiological phenomenon, which has thus far been overlooked. It reflects a robust correction mechanism, which assures that target is reached even if the pre-oculomotor drive through the basal ganglia to the superior colliculus becomes abnormally weak or under inadequately strong inhibition - as is postulated for PD. Thus, only the abnormally high incidence of fragmentation, and of the associated amplitude reduction of the primary saccades, rather than the phenomenon per se, can be used as a diagnostic criterion in early stages of PD.

Aged↗
PubMed2006

Small-scale experimental habitat fragmentation reduces colonization rates in species-rich grasslands.

Habitat fragmentation is one of the most important threats to biodiversity. Decreasing patch size may lead to a reduction in the size of populations and to an increased extinction risk of remnant populations. Furthermore, colonization rates may be reduced in isolated patches. To investigate the effects of isolation and patch size on extinction and colonization rates of plant species, calcareous grasslands at three sites in the Swiss Jura Mountains were experimentally fragmented into patches of 0.25, 2.25, and 20.25 m2 by frequent mowing of the surrounding area from 1993 to 1999. Species richness in the fragment plots and adjacent control plots of the same sizes was recorded during these 7 years. In agreement with the theory of island biogeography, colonization rate was reduced by 30% in fragments versus non-isolated controls, and extinction increased in small versus large plots. Habitat specialists, in contrast to generalists, were less likely to invade fragments. In the last 4 years of the experiment, extinction rates tended to be higher in fragment than in control plots at two of the three sites. Despite reduced colonization rates and a tendency of increased extinction rates in fragments, fragmented plots had only marginally fewer species than control plots after 7 years. Hence, rates were a more sensitive measure for community change than changes in species richness per se. From a conservation point of view, the detected reduced colonization rates are particularly problematic in small fragments, which are more likely to suffer from high extinction rates in the long run.

Biodiversity↗
PubMed1990

Purification and characterization of a large, tryptic fragment of human thyroid peroxidase with high catalytic activity.

Thyroid peroxidase (TPO) was purified from human thyroid tissue, obtained at surgery from patients with Graves' disease, by a procedure similar to one that we had previously used for the purification of porcine TPO. The membrane-bound enzyme was solubilized by treatment of the thyroid particulate fraction with trypsin plus detergent. After precipitation with ammonium sulfate, the enzyme was purified by a series of column treatments, including ion-exchange chromatography on DEAE-cellulose, gel filtration through Bio-Gel P-100, and hydroxylapatite chromatography. Although a high degree of purification was achieved, the finally isolated product was considerably more heterogeneous than the TPO obtained from porcine thyroids. Several pools of active enzyme differing in values for A412/A280 and in specific activity were collected. Gel electrophoresis was performed under native, denaturing [sodium dodecyl sulfate (SDS)] and denaturing plus reducing conditions. Native gel electrophoresis indicated that the active enzyme (93 kDa) was heavily contaminated with an inactive 60-kDa fragment, which we were unable to remove by HPLC. The inactive fragment was highly antigenic when tested on immunoblots with an antibody to TPO. The presence of the inactive fragment greatly reduced values for A412/A280 in the finally purified human TPO. Two of the pools, with A412/A280 values of 0.159 and 0.273, were used for further testing. Catalytic activity was very similar in these two pools when measured on the basis of heme content by several different assays. Moreover, the specific activities of both, based on heme content, were very similar to those observed with a porcine TPO preparation with A412/A280 = 0.48. These findings indicate that the inactive 60-kDa fragment most likely did not contain heme. On SDS-polyacrylamide gel electrophoresis under reducing conditions, the 60-kDa fragment completely disappeared and was replaced by a 36- and a 24-kDa component. Amino terminal sequence information obtained on these components indicated that the 24-kDa component represents the amino terminal portion of the active 93-kDa fragment, whereas the 36-kDa fragment represents the carboxyl terminal portion. A model is proposed suggesting that the 60-kDa fragment was generated by trypsin cleavage of native TPO at two internal sites within a disulfide loop (res approximately 300 and res 564) and at one further internal site (res 280). In addition, trypsin cleavage is proposed at sites near the amino and carboxyl ends common to both the active 93-kDa and the inactive 60-kDa fragments.(ABSTRACT TRUNCATED AT 400 WORDS)

Amino Acid Sequence↗
PubMed1990

Gallbladder stone fragments in feces after biliary extracorporeal shock-wave lithotripsy.

All stools passed on the first 3 days after extracorporeal shock-wave lithotripsy of gallbladder stones in 21 patients were collected and examined for the presence of stone fragments. A total of 555 fragments varying in number per patient (4-69) and in size (maximum diameters from 0.5-8.0 mm) were recovered by sieving aqueous suspensions of the feces. All 482 fragments less than or equal to 3.0 mm left the biliary tract without any clinical symptoms, as did the three largest fragments with maximum diameters of 7.0-8.0 mm and almost all of the 70 fragments measuring 3.5-5.0 mm. During the observation period, four episodes of biliary complaints were recorded in three patients in whom fragments with maximum diameters of 3.5-5.0 mm were found. The only chemical abnormality was a temporary elevation of lipase activity to twice the normal range in 1 case. All fragments were identified as gallbladder stones by infrared spectroscopy on the basis of their (varyingly high) cholesterol content. By macroscopic criteria, most of the fragments were from mixed stones; therefore, provided there is a functioning gallbladder and sufficiently fine fragmentation, successful extracorporeal shock-wave lithotripsy does not seem to be limited to pure cholesterol stones.

Abdominal Pain↗
PubMed1995

The thermodynamics of association and unfolding of the 205-316 C-terminal fragment of thermolysin.

The 205-316 C-terminal fragment of thermolysin has been studied by differential scanning calorimetry at pH values 2.5, 3.0, 3.5, 4.0 and 5.0 and at a constant ionic strength of 130 mM. The thermal unfolding of the fragment occurs at thermodynamic equilibrium under our experimental conditions. The effect of sample concentration at the different pH values on the calorimetric traces is consistent with a monomer-dimer equilibrium of the folded fragment, which undergoes thermal unfolding into individual fragments. Equilibrium sedimentation experiments at 10 degrees C and different pH values confirm the presence of the association equilibrium and provide the value of the dimerization constants. The global analysis of the calorimetric, heat capacity curves has been carried out by a multidimensional fitting to the model N2<-->2N<-->2U. The analysis leads to a complete thermodynamic characterization of both the association and unfolding processes of the fragment. The resulting thermodynamic functions suggest a partially unfolded structure for both the monomeric and dimeric fragment, as well as a conformational change linked to the association process. Our results are discussed in terms of the structural information currently available and compared with the energetics of unfolding of the shorter 255-316 dimeric C-terminal fragment of thermolysin (Conejero-Lara, F., De Filippis, V., Fontana, A. and Mateo, P.L. (1994) FEBS Lett. 344, 154-156). The presence of the additional 50 residues increases the relative population of the 205-316 monomeric fragment versus that of the 255-316 fragment.

Calorimetry, Differential Scanning↗
PubMed1991

Lipopolysaccharide induces double-stranded DNA fragmentation in mouse thymus: protective effect of zinc pretreatment.

Intraperitoneal injection of female NAW/W1 mice with 5 mg of Salmonella typhimurium lipopolysaccharide/kg results in decreased body and thymus weight. Reduced thymic weight is accompanied by fragmentation of DNA into multimers of about 200 bp size. This effect is consistent with the induction of intranucleosomal cleavage of double-stranded DNA in thymus. Maximal fragmentation of DNA occurs between 18 and 24 h after treatment; by 48 h post lipopolysaccharide treatment, there is little evidence of thymic DNA fragmentation. Pretreatment of mice with Zn protects against lipopolysaccharide-induced DNA fragmentation. This effect is maximal at about 72 h after Zn treatment (24 h after lipopolysaccharide treatment) and persists until about 96 h after Zn treatment. At 72 h after pretreatment, the antagonism of thymic DNA fragmentation by Zn is dose-dependent. To examine the role of the acute phase inflammatory response elicited by lipopolysaccharide treatment in the production of changes in thymic weight and DNA integrity, the effects of treatment with casein, a well-characterized inducer of the acute phase inflammatory response in mice, were examined. In contrast to the effect of lipopolysaccharide, casein treatment did not produce a similar pattern of DNA fragmentation in thymus. Taken together, these data suggest that lipopolysaccharide induces DNA fragmentation in thymus by a mechanism which does not occur during the pathophysiological changes which accompany the casein-induced acute phase response. Further, the antagonism by Zn of lipopolysaccharide-induced fragmentation of thymic DNA is consistent with earlier findings that Zn can prevent dexamethasone-induced DNA fragmentation in vitro.

Acute-Phase Reaction↗
PubMed1980

Cloning of avian tumor virus DNA fragments in plasmid pBR322: evidence for efficient transcription in E. coli from a virus-coded promoter.

Two avian tumor virus DNA fragments of 4.2 and 3.2 kb were inserted into pBR322 in the two possible orientations for each fragment. In the Escherichia coli host cells, RNA polymerase initiates transcription of large quantities (up to 0.5 to 1% of total E. coli RNA) of virus-specific RNA in the recombinant plasmids carrying the 4.2-kb fragment (pATV-6) but not in pATV-2 which contains the 3.2-kb fragment. Two SacI cleavage sites flank the putative promoter in the 4.2-kb viral insert. Deletion in the 1.2-kb SacI fragment obliterated the ability of pATV-6 to synthesize viral RNA. Digestion of the 1.2-kb SacI fragment with PvuI generates two fragments of 0.63 and 0.57 kb. Deletion of the 0.57-kb but not the 0.63-kb PvuI-SacI fragment completely eliminated the ability of the recombinant to synthesize viral RNA. These results strongly suggest that viral RNA in E. coli transcription is indeed initiated at a size present in the viral genome and that this site is localized in the 0.57-kb PvuI-SacI fragment.

Cloning, Molecular↗
PubMed1982

A new pair of M13 vectors for selecting either DNA strand of double-digest restriction fragments.

The strategy of shotgun cloning with M13 is based on obtaining random fragments used for the rapid accumulation of sequence data. A strategy, however, is sometimes needed for obtaining subcloned sequences preferentially out of a mixture of fragments. Shotgun sequencing experiments have shown that not all DNA fragments are obtained with the same frequency and that the redundant information increases during the last third of a sequencing project. In addition, experiments have shown that particular fragments are obtained more frequently in one orientation, allowing the use of only one of the two DNA strands as a template for M13 shotgun sequencing. Two new M13 vectors, M13mp8 and M13mp9, have been constructed that permit the cloning of the same restriction fragment in both possible orientations. Consequently, each of the two strands becomes a (+) strand in a pair of vectors. The fragments to be cloned are cleaved with two restriction enzymes to produce a fragment with two different ends. The insertion of such a fragment into the vector can occur only in one orientation. Since M13mp8 and M13mp9 have their array of cloning sites in an antiparallel order, either orientation for inserting a double-digest fragment can be selected by the choice of the vector.

Amino Acid Sequence↗
PubMed1982

Foot-and-mouth disease virus: immunogenicity and structure of fragments derived from capsid protein VP and of virus containing cleaved VP.

Peptide fragments were obtained from the immunogenic capsid protein VP3, ca. 24 kilodaltons (kd), of foot-and-mouth disease virus type A12 119ab by three procedures: (1) spontaneous proteolysis of in virion VP3 in tissue cultures to produce a 15 kd peptide, designated S fragment; (2) trypsin treatment of purified virus to produce a 16 kg peptide, designated T fragment; and (3) cyanogen bromide cleavage of purified VP3 to produce a 13 kd fragment. Following isolation and purification by gel electrophoresis, VP3 and each of the three fragments were immunogenic for livestock. Lyophilization appeared to impair the immunogenicity of VP3. In addition, viruses containing VP3 fragments produced either by the spontaneous- or trypsin-induced proteolysis were as immunogenic as virus with its VP3 intact. Amino acid sequencing of N-terminal regions revealed that the S fragment was homologous with the N-terminus of VP3, whereas the 13 kd fragment possessed a unique N-terminus. Thus, putative common immunogenic amino acid sequences would appear to reside within an overlap region of the 15 kd S and 13 kd fragments. Sequencing of cDNA prepared to viral genome RNA provided three kinds of information: it (1) placed the above overlap region in the second and third quarters of VP3; (2) demonstrated that the codons for the C-terminus of VP1 and N-terminus of VP3 are contiguous; and (3) supported earlier evidence that these same codons program a chain reversal where VP1 and VP3 are joined in the precursor polyprotein.

Amino Acid Sequence↗
PubMed2005

Factors affecting SFHR gene correction efficiency with single-stranded DNA fragment.

A 606-nt single-stranded (ss) DNA fragment, prepared by restriction enzyme digestion of ss phagemid DNA, improves the gene correction efficiency by 12-fold as compared with a PCR fragment, which is the conventional type of fragment used in the small fragment homologous replacement method [H. Tsuchiya, H. Harashima, H. Kamiya, Increased SFHR gene correction efficiency with sense single-stranded DNA, J. Gene Med. 7 (2005) 486-493]. To reveal the characteristic features of this gene correction with the ss DNA fragment, the effects on the gene correction in CHO-K1 cells of the chain length, 5'-phosphate, adenine methylation, and transcription were studied. Moreover, the possibility that the ss DNA fragment is integrated into the target DNA was examined with a radioactively labeled ss DNA fragment. The presence of methylated adenine, but not the 5'-phosphate, enhanced the gene correction efficiency, and the optimal length of the ss DNA fragment (approximately 600 nt) was determined. Transcription of the target gene did not affect the gene correction efficiency. In addition, the target DNA recovered from the transfected CHO-K1 cells was radioactive. The results obtained in this study indicate that length and adenine methylation were important factors affecting the gene correction efficiency, and that the ss DNA fragment was integrated into the double-stranded target DNA.

Adenine↗
PubMed2005

Hydrodynamic fragmentation of nanoparticle aggregates at orthokinetic coagulation.

Hydrodynamic forces on a doublet of large particles or aggregates during sedimentation cause the non-inertial fragmentation of the doublet, if the doublet Reynolds number and Stokes number are small. In lio-dispersed systems, this non-inertial fragmentation is known to promote the wet classification of large particles whereas the small particles are aggregated (in the secondary minimum) and hence cannot be separated despite the electrostatic repulsion. In aero-dispersed systems with negligible electrostatic repulsion, it is possible to have a narrower separation between two interacting particles so that the attractive surface forces such as van der Waals force could increase in orders of magnitude. As a result, the doublet fragmentation by the aerodynamic detaching force becomes very difficult or even impossible in sedimentation at a small Re. However, this study shows that, when extending above analysis for the interacting fractal aggregates of nanoparticles in a suspension, it is still possible to have aggregate (doublet of two nano-aggregates) fragmentation by the aerodynamic detaching force because the surface forces for nanoparticle contact between two aggregates may be in orders of magnitude smaller than that for micron-sized particles. Even with multiple contacts between two interacting nanoparticle aggregates, this prediction of aggregate fragmentation may still be valid because the contacts may break step by step due to the aggregate rolling along each other caused by a short-range aerodynamic interaction during their differential settling. The aerodynamics of aero-dispersed nanoparticle aggregates is analogous to the hydrodynamics of lio-dispersed solid particles. Therefore the hydrodynamic fragmentation model may be used to partially interpret the stability of nanoparticle fluidization process. Our model indicates that, without continued doublet fragmentation of nano-aggregates, the fluidized nanoparticle suspension would be de-fluidized within minutes via the cascading aggregation. However, the experimental evidence of a sustained operation of fluidization of nanoparticle aggregates over a very long time period without changing the size of nanoparticle aggregates indicates that there is a dynamic balance between the aggregation and fragmentation of nano-aggregates in nanoparticle fluidization. The prediction of critical size of nanoparticle aggregates caused by fragmentation in stabilized fluidization agrees with our in situ measurements.

Kinetics↗