Rapid detection of sequence polymorphisms in the human mitochondrial DNA control region by polymerase chain reaction and single-strand conformation analysis in mutation detection enhancement gels

The article describes a rapid approach for the detection of sequence polymorphisms in the mitochondrial (mt)DNA control region that involves enzymatic amplification of each entire mtDNA control region (HV1 and HV2) and the subsequent analysis of the PCR products by single-strand conformation analysis (SSCA) in mutation detection enhancement (MDE) gels, followed by silver stain detection. HV1 and HV2 SSC reference ladders were developed to standardize the classification of the different mtDNA types. Twenty-five mtDNA types were observed among the 45 Spanish individuals analyzed: 11 types were observed in the HV1 region as compared with 10 types in the HV2 region. This mutation scanning strategy could be a promising method of potential use not only in forensic genetics but also in population and evolutionary studies.     

Analysis of mouse glomerular podocyte mRNA by single-cell reverse transcription-polymerase chain reaction

The polymorphic control region of mitochondrial DNA (mtDNA) is becoming more commonly used in forensic applications to differentiate among individuals in a population. Two hypervariable regions (HV1 and HV2) are often sequenced following amplification of the mtDNA via the polymerase chain reaction (PCR). More rapid screening assays would reduce both the effort and the expense of comparing two samples. A methodology has been developed that first uses restriction endonuclease digestion of the PCR-amplified mtDNA using RsaI and MnlI and then capillary electrophoresis (CE) to separate and size the PCR-RFLP fragments. This rapid procedure offers an alternative method for screening of polymorphisms in amplified mtDNA samples. In addition, the presence of a T-->C transition at position 16189, which gives rise to the so-called "C-stretch" in HV1, may be predicted from the presence of nonspecific PCR products in the CE results.     

Mitochondrial DNA polymorphism in Jindo dogs

Mitochondrial DNA (mtDNA) polymorphism was studied in 21 Jindo dogs inhabiting Jin Island off the Korean peninsula. The polymorphism was analyzed with 10 restriction endonucleases that recognize six base pairs. The sizes of the mtDNA fragments produced by digestion using each endonucleases were separated by agarose gel electrophoresis, and the polymorphisms were detected with Japanese mongrel dog mtDNA as a probe. The mtDNA polymorphism in Jindo dogs was observed with four restriction endonucleases, Apa I, EcoR V, Hinc II, and Sty I. However, no polymorphism was detected with BamH I, Bgl II, EcoR I, Hind III, Pst I, or Xba I. The observed restriction endonuclease morphs were classified into 4 types of distinct cleavage patterns. The average number of nucleotide substitutions per nucleotide site in Jindodogs was estimated to be 0.0086. By UPG phylogenetic analysis, the 4 mtDNA types showed only one cluster. This suggests that Jindo dogs have not diverged from the other cluster up to the present and the species is considerably pure.     

Sequence polymorphism of mitochondrial DNA control region in Japanese

Sequence polymorphisms of the mitochondrial DNA (mtDNA) control region, hypervariable regions I and II, from 100 unrelated Japanese were determined by PCR amplification and direct sequencing. Sequences of 404 nucleotides for hypervariable region I and 379 nucleotides for region II were obtained. Variable sites (85 and 45) were revealed in region I and region II, respectively, as compared to the reference sequence, and a total of 96 different genetic patterns from both regions I and II were determined. A point mutation heteroplasmy was observed at the ratio of approximately 50:50 from one individual at the sequence position 151 showing a nucleotide transition from C to T. The probability of identity was estimated as 2.3% for region I, 3.9% for region II, and 1.1% combined for both regions. These results suggest that sequence polymorphism of mtDNA control region would be very useful in forensic practice as a marker for individual identification.     

Differences in familism values and caregiving outcomes among Korean, Korean American, and White American dementia caregivers.

Recent theories have suggested that burden and distress among dementia caregivers may be higher in American culture, which emphasizes individualism, and lower in cultures with higher levels of familism. However, immigrants may experience higher levels of burden because of acculturation with attendant values, conflicts and stresses. Forty-four Korean caregivers and 32 Korean American caregivers were compared with 54 White American caregivers on sociodemographic variables, familism, burden, anxiety, and depression. Familism was highest in Korean caregivers and lowest in Whites, with Korean Americans in the middle. Koreans and Korean Americans reported higher levels of burden. Koreans showed higher levels of depression and of anxiety than White American caregivers, with Koreans and Korean Americans higher than Whites on anxiety. These results suggest a need for greater specificity in theories about familism values, with attention to the specific meaning of familism in different cultures. (PsycINFO Database Record (c) 2010 APA, all rights reserved)     

The J. David Gladstone Institutes

This study was designed to examine the relation between pulpal nerves and the differentiation of pulpal cells into preodontoblasts and odontoblasts during the healing process after pulpotomy. A total of 36 upper and lower teeth obtained from six adult dogs were used. The pulp chamber was opened with a sterile diamond bur, the coronal pulp was exposed, and the whole surface of the amputated pulp was capped with calcium hydroxide. The interval between pulpotomy and extraction was 5, 7, 10, 15, 20, 30, 40, 50, and 60 days, and then specimens were examined ultrastructurally. Close contact between fibroblast-like cells/osteoblast-like cells and nerve terminals at the calcification front was observed in the early healing process after pulpotomy, suggesting a close relation between nerve fibers and pulpal cell differentiation.     

Diet and risk of colon cancer in a large prospective study of older women: an analysis stratified on family history (Iowa, United States)

The aim of this study was to determine if diffusible angiogenic growth factors were released in human dental pulp during orthodontic tooth movement. These factors, if diffusible, could induce angiogenesis in other tissues, and may then be isolated and identified. The pulps from 14 premolar teeth treated with straight wire fixed orthodontic appliances for 2 weeks were compared with those of 14 untreated control premolar teeth from the same subjects. Following tooth extraction and sectioning, 1-mm horizontal sections of pulp tissue were embedded in collagen with 1-mm sections of rat aorta and co-cultured in growth media for up to 4 weeks. Sections of rat aorta alone were also cultured. Angiogenic changes in the form of microvessel growth were observed by light microscopy. Microvessel identification was confirmed by electron microscopy and by immunohistochemistry using staining for factor VIII-related antigen marker for endothelial cells. When compared at days 5, 10, and 14 of co-culture, the number of microvessels was significantly greater in the pulps from orthodontically moved teeth than in those from the control teeth. The number of rat aorta microvessels was also significantly greater when co-cultured with pulp from orthodontically moved teeth than with pulp from control teeth and when compared with control cultures of rat aorta alone. There were no significant differences in microvessel numbers between the rat aorta co-cultured with pulp from control teeth and control cultures of rat aorta alone. These results indicate an increase in angiogenic growth factors in the pulp of orthodontically moved teeth, and the enhanced response of the rat aorta when co-cultured with this pulp shows that these factors appear to be diffusible.     

Mutations in mitochondrial DNA accumulate differentially in three different human tissues during ageing

In 60 human tissue samples (encompassing skeletal muscle, heart and kidney) obtained from subjects aged from under 1 to 90 years, we used quantitative PCR procedures to quantify mitochondrial DNA (mtDNA) molecules carrying the 4977 bp deletion (mtDNA4977) and 3243 A-->G base substitution. In addition, the prevalence of multiple mtDNA deletions was assessed in a semi-quantitative manner. For all three tissues, the correlations between the accumulation of the particular mtDNA mutations and age of the subject are highly significant. However, differential extents of accumulation of the two specific mutations in the various tissues were observed. Thus, the mean abundance (percentage of mutant mtDNA out of total mtDNA) of mtDNA4977in a subset of age-matched adults is substantially higher in skeletal muscle than in heart and kidney. However, the mean abundance of the 3243 A-->G mutation in skeletal muscle was found to be lower than that in heart and kidney. Visualisation of arrays of PCR products arising from multiple mtDNA deletions in DNA extracted from adult skeletal muscle, was readily made after 30 cycles of PCR. By contrast, in DNA extracted from adult heart or kidney, amplification for 35 cycles of PCR was required to detect multiple mtDNA deletions. Although such multiple deletions are less abundant in heart and kidney than in skeletal muscle, in all tissue extracts there are unique patterns of bands, even from different tissues of the same subject. The differential accumulation of mtDNA4977, other mtDNA deletions and the 3243 A-->G mutation in the three tissues analysed presumably reflects different metabolic and senescence characteristics of these various tissues.     

Ultrasound biomicroscopy of the anterior segment of the eyes of infants

Multiplex PCR amplification has been useful for gene mapping with polymorphic short tandem repeat (STR) loci. We have tested the four loci D20S470, D13S325, HumFOLP23 and D10S2325 for the simultaneous typing of more than 100 unrelated Koreans. This analysis allows a single base pair resolution and rapid typing with silver staining. The allele and genotype distributions are in accordance with Hardy - Weinberg expectations. These STR loci have proven useful for forensic analysis and paternity tests in which the variable number of tandem repeat (VNTR) loci have some limitations.     

A human mitochondrial DNA standard reference material for quality control in forensic identification, medical diagnosis, and mutation detection

A human mitochondrial DNA (mtDNA) standard reference material (SRM 2392) will provide quality control when mtDNA is sequenced for forensic identifications, medical diagnosis, or mutation detection. SRM 2392 includes DNA from two lymphoblast cell cultures (CHR and 9947A) and cloned DNA from the CHR HV1 region, which contains a C stretch and is difficult to sequence. The mtDNA sequence (but not the DNA) of a third human template GM03798 is provided for comparison. Fifty-eight unique primer sets allow any area or the entire mtDNA (16,569 bp) to be amplified and sequenced. While none of the differences in these three templates correspond to published mutations associated with specific diseases, some of these differences did result in animo acid changes compared with that published by S. Anderson et al. (1981, Nature 290: 457-465). An interlaboratory evaluation of the amplification, sequencing, and data analysis of the CHR template was conducted by four laboratories. Corroboration of the SRM results will provide quality assurance that any unknown mtDNA is also being amplified and sequenced correctly.     

An evaluation of introgression of Atlantic coast striped bass mitochondrial DNA in a Gulf of Mexico population using formalin-preserved museum collections

Striped bass Morone saxatilis populations in drainages along the Gulf of Mexico coast (Gulf) were depleted in the 1950s and 1960s, probably because of anthropogenic influences. It is believed that only the Apalachicola-Chattahoochee-Flint (A-C-F) river system continually supported a naturally reproducing population of Gulf lineage. Striped bass juveniles of Atlantic coast (Atlantic) ancestry were introduced to restore population abundances in the A-C-F from the late 1960s to the mid 1970s and in many other Gulf rivers from the 1960s to the present. We previously identified mtDNA polymorphisms that were unique to approximately 60% of striped bass from the A-C-F and which confirmed the continued successful natural reproduction of striped bass of Gulf maternal ancestry within the system. However, the genetic relatedness of the extant A-C-F population to 'pure' Gulf striped bass was not addressed. In this study, we determined the frequency of a diagnostic mtDNA XbaI polymorphism in samples of 'pure' Gulf striped bass that were collected from the A-C-F prior to the introduction of Atlantic fish, that were obtained from museum collections, and that were originally preserved in formalin. PCR primers were developed that allowed for amplification of a 191-bp mtDNA fragment that contained the diagnostic XbaI restriction site. Using RFLP and direct sequence analyses of the PCR amplicons, we found no significant differences in mtDNA XbaI genotype frequencies between the archived samples and extant A-C-F samples collected over a 15-year period. This indicates that significant maternally mediated introgression of Atlantic mtDNA genomes into the A-C-F gene pool has not occurred. Additionally, we found no evidence of the unique Gulf mtDNA genotype in striped bass from extant populations in Texas, Louisiana and the Mississippi River. These results highlight the importance of the A-C-F as a repository of striped bass to restore extirpated Gulf populations and the potential use of museum collections in retrospective population studies.     

Replicating the positivity effect in picture memory in Koreans: Evidence for cross-cultural generalizability.

Older adults’ relatively better memory for positive over negative material (positivity effect) has been widely observed in Western samples. This study examined whether a relative preference for positive over negative material is also observed in older Koreans. Younger and older Korean participants viewed images from the International Affective Picture System (IAPS), were tested for recall and recognition of the images, and rated the images for valence. Cultural differences in the valence ratings of images emerged. Once considered, the relative preference for positive over negative material in memory observed in older Koreans was indistinguishable from that observed previously in older Americans. (PsycINFO Database Record (c) 2010 APA, all rights reserved)     

Effectiveness of caries removal by the partial tunnel preparation method

The tunnel preparation method is designed to remove approximal caries through a channel from the occlusal surface while preserving the marginal ridge. This method entails reduced access to the caries lesion and thereby uncertainty as to the complete removal of caries. The purpose of the present investigation was to study the effectiveness of caries removal in 60 extracted premolars and molars by the partial tunnel preparation method. The glass polyalkenoat (ionomer) filling and the distance to the pulp were also examined. Examination of the sectioned teeth showed residual caries in the axial wall of two teeth and in dentin close to the enamel lesion in 10 teeth. Very few porosities were found within the glass polyalkenoat material and at the interface between the filling and the cavity walls.     

Serum amyloid A induces chemotaxis of human mast cells by activating a pertussis toxin-sensitive signal transduction pathway

Human MitBASE is a database collecting human mtDNA variants. This database is part of a greater mitochondrial genome database (MitBASE) funded within the EU Biotech Program. The present paper reports the recent improvements in data structure, data quality and data quantity. As far as the database structure is concerned it is now fully designed and implemented. Based on the previously described structure some changes have been made to optimise both data input and data quality. Cross-references with other bio-databases (EMBL, OMIM, MEDLINE) have been implemented. Human MitBASE data can be queried with the MitBASE Simple Query System (http://www.ebi.ac.uk/htbin/Mitbase/mit base.pl) and with SRS at the EBI under the 'Mutation' section (http://srs.ebi.ac.uk/srs5/). At present the HumanMitBASE node contains approximately 5000 variants related to studies investigating population polymorphisms and pathologies.     

搅拌摩擦加工IF钢的组织性能

对3 mm厚的DC04冷轧IF钢板进行搅拌摩擦加工,研究加工区域的微观组织与力学性能.在旋转速度为950 r·min^-1,加工速度为60 mm·min^-1时,采用加工后强制冷却技术可获得光滑平整且没有缺陷的加工表面.搅拌摩擦加工后组织显著细化,加工中心的平均显微硬度约为HV 135.6,是母材硬度的1.4倍,表面细晶层硬度最高可达到HV 312.8,细晶层和过渡层的抗拉强度分别比母材的抗拉强度提高50.9%和47.6%,加工前后试样的拉伸断口均呈微孔聚合韧性断裂特征.细晶强化对材料抗拉强度的提高起主要作用.     

Use of dental instruments for bridging during electric pulp testing

It has been suggested that dental instruments can be used as bridging instruments to facilitate electric pulp testing of teeth with extensive restorations. This study reports a clinical investigation to evaluate the effectiveness of this procedure. One hundred seventeen vital teeth in 20 volunteers were tested. Ten endodontically treated teeth functioned as controls. Following appropriate isolation and asepsis technique, baseline recordings of the threshold response with the electric pulp tester were taken. With the use of dental explorers and endodontic files to bridge between the probe tip and the tooth surface, recordings were made of the threshold responses. Findings indicate that electrically conductive dental instruments can be reliably used as bridging instruments with the electric pulp tester.     

Ancient mtDNA sequences in the human nuclear genome: a potential source of errors in identifying pathogenic mutations

Nuclear-localized mtDNA pseudogenes might explain a recent report describing a heteroplasmic mtDNA molecule containing five linked missense mutations dispersed over the contiguous mtDNA CO1 and CO2 genes in Alzheimer's disease (AD) patients. To test this hypothesis, we have used the PCR primers utilized in the original report to amplify CO1 and CO2 sequences from two independent rho degrees (mtDNA-less) cell lines. CO1 and CO2 sequences amplified from both of the rho degrees cells, demonstrating that these sequences are also present in the human nuclear DNA. The nuclear pseudogene CO1 and CO2 sequences were then tested for each of the five "AD" missense mutations by restriction endonuclease site variant assays. All five mutations were found in the nuclear CO1 and CO2 PCR products from rho degrees cells, but none were found in the PCR products obtained from cells with normal mtDNA. Moreover, when the overlapping nuclear CO1 and CO2 PCR products were cloned and sequenced, all five missense mutations were found, as well as a linked synonymous mutation. Unlike the findings in the original report, an additional 32 base substitutions were found, including two in adjacent tRNAs and a two base pair deletion in the CO2 gene. Phylogenetic analysis of the nuclear CO1 and CO2 sequences revealed that they diverged from modern human mtDNAs early in hominid evolution about 770,000 years before present. These data would be consistent with the interpretation that the missense mutations proposed to cause AD may be the product of ancient mtDNA variants preserved as nuclear pseudogenes.     

Reliability of electric pulp testing after pulpal testing with dichlorodifluoromethane

The purpose of this study was to determine if the sequence and interval between electric pulp testing and cold vitality testing with dichlorodifluoromethane affects the reliability of pulpal diagnostic testing. Sixty vital teeth in 15 volunteers were tested. Ten endodontically treated teeth were used as negative controls. After isolation and asepsis techniques, baseline threshold responses from a digital electric pulp tester were recorded from the maxillary incisors. A dichlorodifluoromethane-saturated cotton pellet was applied to teeth 8, 9, and 10. Electric pulp testing was repeated at 30-s, 1-min, and 2-min intervals on all test teeth after the cold test. The level of responses were recorded and statistically analyzed. The results of this study indicate that electric pulp testing is not adversely affected by the use of dichlorodifluoromethane.     

European Y-chromosomal lineages in Polynesians: a contrast to the population structure revealed by mtDNA

We have used Y-chromosomal polymorphisms to trace paternal lineages in Polynesians by use of samples previously typed for mtDNA variants. A genealogical approach utilizing hierarchical analysis of eight rare-event biallelic polymorphisms, seven microsatellite loci, and internal structural analysis of the hypervariable minisatellite, MSY1, has been used to define three major paternal-lineage clusters in Polynesians. Two of these clusters, both defined by novel MSY1 modular structures and representing 55% of the Polynesians studied, are also found in coastal Papua New Guinea. Reduced Polynesian diversity, relative to that in Melanesians, is illustrated by the presence of several examples of identical MSY1 codes and microsatellite haplotypes within these lineage clusters in Polynesians. The complete lack of Y chromosomes having the M4 base substitution in Polynesians, despite their prevalence (64%) in Melanesians, may also be a result of the multiple bottleneck events during the colonization of this region of the world. The origin of the M4 mutation has been dated by use of two independent methods based on microsatellite-haplotype and minisatellite-code diversity. Because of the wide confidence limits on the mutation rates of these loci, the M4 mutation cannot be conclusively dated relative to the colonization of Polynesia, 3,000 years ago. The other major lineage cluster found in Polynesians, defined by a base substitution at the 92R7 locus, represents 27% of the Polynesians studied and, most probably, originates in Europe. This is the first Y-chromosomal evidence of major European admixture with indigenous Polynesian populations and contrasts sharply with the picture given by mtDNA evidence.     

Culture and judgement of causal relevance.

The authors hypothesized that because the causal theories of East Asians were more holistic and complex than those of Americans, the amount of information considered before making a final attribution would be larger for East Asians than for Americans. This hypothesis was supported through 4 studies. When participants attempted to explain a deviant behavior (Study 1) or a prosocial behavior (Study 2), Korean participants took into consideration a greater amount of information than did either American or Asian American participants. Study 3 replicated the findings of Studies 1 and 2 within each culture. Finally, Study 4 established a link between the present findings and past research on culture and attribution. Namely, Study 4 found that Koreans made more external attributions than Americans because Koreans considered more information than did Americans. (PsycINFO Database Record (c) 2010 APA, all rights reserved)