VWA基因座在一个三代家系发生的两次突变

目的探讨短串联重复序列遗传变异的方式。方法用Identifiler^TM试剂盒和DNATyper15^TM试剂盒进行检测,用变性聚丙烯酰胺凝胶电泳法对VWA基因座突变家系中3代父、母、子的DNA样本进行STR基因分型,将需测序的等位基因条带从凝胶上切下,再进行PCR扩增,产物经纯化后进行测序。结果此3代家系中Ⅱ-7、Ⅲ-8发生等位基因变异表现为一个重复单位的增加或减少。其中Ⅱ-7的一条发生突变的等位基因具体来源可知,Ⅲ-8的一条突变等位基因无法确定。结论本文中VWA基因座等位基因突变主要表现为一个重复单位的增加,没有碱基的插入或缺失,突变主要来自父亲,突变等位基因无TCTA（TCTG）3（TCTA）n、TCTA（TCTG）5-6（TCTA）n突变类型,均为TCTA（TCTG）4（TCTA）n型突变。     

Antigenic properties of human and animal bloodstains studied by enzyme-linked immunosorbent assay (ELISA) using various antisera against specific plasma proteins

Antigenic properties of bloodstains of human and non-human primates as well as other animal bloodstains were investigated by the inhibition ELISA using commercially available anti-human albumin (Alb), alpha 2-macroglobulin (alpha 2-M), fibrinogen, transferrin, and immunoglobulin G. In general, chimpanzee bloodstains showed strong cross-reactions with these antisera, and the extent of the cross-reactions of other animal bloodstains decreased largely with the phylogenic order, i.e., agile gibbon (ape), Old World monkeys (Japanese monkey and hamadryas baboon), New World monkeys (night monkey and tufted capuchin monkey), prosimians (grand galago and ring-tailed lemur) and other animals (rat, cattle, swine, goat, dog, cat, and chicken). Among these antisera, anti-human alpha 2-M showed the weakest cross-reaction with chimpanzee bloodstains, and anti-human Alb showed next.     

To Each According to his Need? Variability in the Responses to Inequity in Non-Human Primates

While it is well established that humans respond to inequity, it remains unclear the extent to which this behavior occurs in our non-human primate relatives. By comparing a variety of species, spanning from New World and Old World monkeys to great apes, scientists can begin to answer questions about how the response to inequity evolved, what the function of this response is, and why and how different contexts shape it. In particular, research across non-human primate species suggests that the response is quite variable across species, contexts, and individuals. In this paper, we aim to review these differences in an attempt to identify and better understand the patterns that emerge from the existing data with the goal of developing directions for future research. To begin, we address the importance of considering socio-ecological factors in non-human primates in order to better understand and predict expected patterns of cooperation and aversion to inequity in different species, following which we provide a detailed analysis of the patterns uncovered by these comparisons. Ultimately, we use this synthesis to propose new ideas for research to better understand this response and, hence, the evolution of our own responses to inequity.     

Fairness in Non-human Primates?

Humans have a sense of fairness, i.e. an interest in the ideal of equity. This sense allows them to compare their own efforts and subsequent outcomes with those of others, and thus to evaluate and react to inequity. The question is whether our closest living relatives, the non-human primates, show the behavioural characteristics that might qualify as necessary components to a sense of fairness, such as inequity aversion. In this article, we review the five different experimental approaches to studying behaviours related to fairness in non-human primates, including their underlying logic and main findings that represent the current state of research in this field. In the critical condition of all these studies, a subject and a conspecific partner have either to invest different efforts or receive different outcomes while observing each other. The main question is whether??and how??subjects react to unequal situations that humans would perceive as ??unfair??. Taken together, the results from all five approaches provide only weak evidence for a sense of fairness in non-human primates. Although apes and monkeys are attentive to what the partner is getting, they do not seem to be able or motivated to compare their own efforts and outcomes with those of others at a human level. Even though the debate is still on-going, we believe that a full sense of fairness is not essential for cooperation. Obviously, apes and monkeys are capable of solving problems cooperatively, without a strong, humanlike sense of fairness. They are mainly interested in maximizing their own benefit, regardless of what others may receive. It is thus possible that a sense of fairness only exists rudimentarily in non-human primates.     

Results of the GEP-ISFG collaborative study on two Y-STRs tetraplexes: GEPY I (DYS461, GATA C4, DYS437 and DYS438) and GEPY II (DYS460, GATA A10, GATA H4 and DYS439)

A collaborative exercise was carried out by the Spanish and Portuguese ISFG Working Group (GEP-ISFG) in order to evaluate the performance of two Y-chromosome STR PCR tetraplexes, which include the loci DYS461, GATA C4, DYS437 and DYS438 (GEPY I), and DYS460, GATA A10, GATA H4 and DYS439 (GEPY II). The participating laboratories were asked to type three samples for the eight markers, using a specific amplification protocol. In addition, two control samples, with known haplotypes, were provided. The results obtained by the 13 different participating laboratories were identical, except for two laboratories that failed to type correctly the same two samples for GATA C4. By sequence analyses, two different GATA C4 allele structures were found. One control sample (allele 21) and two questioned samples (allele 22, correctly typed by all the laboratories, and allele 25) presented the following repeat structure: (TCTA)4(TGTA)2(TCTA)2(TGTA)2(TCTA)n, but different from the one found for allele 26 in one sample included in this exercise, as well as in the second control sample (allele 23), namely (TCTA)4(TGTA)2(TCTA)2(TGTA)2(TCTA)2(TGTA)2(TCTA)n. The collaborative exercise results proved that both Y-tetraplexes produce good amplification results, with the advantage of being efficiently typed using different separation and detection methodologies. However, since GATA C4 repeat presents a complex structure, with alleles differing in sequence structure, efficient denaturing conditions should be followed in order to avoid typing errors due to sizing problems.     

Hypervariable locus of the 3'-flanking region of the neurotensin receptor gene: an effective region for personal identification in forensic practice

We examined the complex short tandem repeat (STR) locus at the 3'-flanking region of the neurotensin receptor (NTR) gene. The polymorphism of this locus was first reported as a simple tetranucleotide repeat variation by Le et al., but it also offers a surprisingly informative variation, that permits reliable individual identification by two complementary strategies: fluorescent-labelled polymerase chain reaction (PCR)/electrophoresis and direct sequencing of the PCR products. We determined the alleles in 203 Japanese by fluorescent-labelled PCR/electrophoresis. Determination was based on their length with a reliability of +/-1 bp, and the frequency of each allele was very low. Sequencing analysis further grouped these alleles in detail. Sequencing demonstrated that the locus varied by six repetitive units and three insertion/deletion positions of nucleotide fragments. We detected multiple alleles having different structures even in the same allele length. We found structural differences in homozygous alleles having the same base pair size. We also determined that apparently homozygous alleles were heterozygous from sequencing electropherograms showing an overlap of nucleotides or +/-1 bp difference. These results indicate that this locus is structurally hypervariable in addition to having allelic length variations, promising a great advance in individual identification in forensic practice.     

Empathy and Fairness: Psychological Mechanisms for Eliciting and Maintaining Prosociality and Cooperation in Primates

In the past, prosociality has been considered a hallmark of humans; however, recently, accumulating data have empirically revealed that non-human animals also show prosocial behavior. In situations in which animals cannot predict return benefits, prosocial behavior is probably driven by other-regarding motivation. A sense of fairness and empathy continue to draw attention as the most plausible candidates for the psychological mechanisms underlying such prosocial behavior. In this article, we first introduce comparative studies on prosocial behavior in non-human primates and discuss similarities and differences between humans and non-human primates. Then, we discuss the role of a sense of fairness and empathy. In this paper, we hypothesize that empathy may promote prosocial behavior, whereas a sense of fairness may play a role as a stabilizer, but not as a promoter of prosocial behavior in non-human animals. We further hypothesize that prosocial behavior motivated by sympathetic concerns can survive only with a sense of fairness, the inhibitory system for unnecessarily excessive expression of prosocial behavior. Without a sense of fairness, empathic animals might be exploited by free-riders, which might lead to the extinction of cooperation. Therefore, the interplay of a sense of fairness and empathy are both important to maintaining prosocial behavior and cooperation. This hypothesis seems to be supported by comparative studies with non-human primates and also by neural studies with humans.     

The short tandem repeat locus VWF2 in Intron 40 of the von Willebrand factor gene consists of two polymorphic sub-loci

This study describes the complex nucleotide sequence structure of the TCTA short tandem repeat (STR) locus, VWF2. Eight alleles of VWF2 were observed in a population of 116 unrelated Caucasian individuals. The alleles ranged in size from 150 to 178 base pairs (bp). Sequence analysis of the isolated alleles revealed two polymorphic regions that were named sub-loci VWF2-a and VWF2-b. VWF2-a is located at the 5' end of the conventional locus, whilst VWF2-b is located at the 3' end. The two sub-loci are joined by a 30-nucleotide non-polymorphic sequence which contains two additional TCTA motif repeats. A semi-nested polymerase chain reaction (PCR) was designed to amplify the VWF2-b region in conjunction with the standard VWF2 amplification. This new amplification method enabled a higher level of allele discrimination than could be achieved by assigning alleles according to size. A cohort of 99 unrelated individuals was tested with this method. VWF2-a expressed five different alleles ranging from zero motif repeats to four motif repeats, while VWF2-b alleles ranged from 8 to 14 motif repeats. Allelic configuration based on the VWF2-a and VWF2-b sub-alleles revealed 23 unique configurations out of a possible 31 for the original eight VWF2 alleles. In conclusion, the VWF2 is a highly polymorphic STR locus with potential application for forensic and parentage testing.     

Identification of DNA of human origin based on amplification of human-specific mitochondrial cytochrome b region

Species-specific differences in a non-polymorphic region of the mitochondrial cytochrome b gene appear to be large enough to allow human-specific amplification of forensic DNA samples. We therefore developed a PCR-based method using newly designed primers to amplify a 157-bp portion of the human mitochondrial cytochrome b gene. The forward and reverse primers were designed to hybridize to regions of the human mitochondrial cytochrome b gene with sequences differing from those of chimpanzee by 26% (7 bp/27 bp) and 26% (6 bp/23 bp), respectively. Using this primer pair, we successfully amplified DNA extracted from blood samples of 48 healthy adults. All these human samples produced a single band of the expected size on agarose gel electrophoresis, and the sequence of the single band was shown to be identical to that of the target region (157 bp) by sequence analysis. On the other hand, no visible bands were amplified from DNA extracted from blood samples of animals including non-human primates (chimpanzee, gorilla, Japanese monkey, crab-eating monkey) and other species (cow, pig, dog, goat, rat, chicken and tuna). Thus, DNA producing a single band following PCR amplification using this primer pair can be reasonably interpreted as being of human origin. In addition, aged biological specimens comprising bloodstains, hair shafts and bones were successfully identified as being of human origin, illustrating the applicability of the present method to forensic specimens.     

The structure, frequency, and forensic application of the STR locus D16S543 in the Japanese population

D16S543 is a complex STR locus consisting of five types of repeat units. The frequency distribution and genetic characteristics of this locus in Japanese were investigated using blood samples from 124 unrelated Japanese and 15 families. Alleles were detected using denatured polyacrylamide gels followed by automated analysis on an ABI 373 sequencer using Genescan software 672. Twenty-one alleles were identified, ranging in size from 281 to 489 bp. An allelic ladder containing the 21 alleles was constructed and used as a typing standard. The repeat unit arrays allowed the 21 alleles to be classified into three distinct groups, including alleles 1 to 7 in group I, alleles 8 to 14 in group II, and alleles 15 to 22 in group III. The alleles in group II were characterized by the insertion of one repeat unit of CAGG, one of AAAG, and three of AAGG, while the group III alleles differed from those of groups I and II by the insertion of a total of 32 repeat units ranging in 5 types. Within each group, the alleles differed from each other only in one 5' side tetranucleotide AAGG. The power of discrimination (Pd) and the estimated heterozygosity were calculated to be 0.989 and 0.934, respectively. Typing of this locus was successfully applied in four old forensic materials. The study presented herein demonstrates that D16S543 is a highly polymorphic and applicable locus in Japanese.     

Sequence variation in humans and other primates at six short tandem repeat loci used in forensic identity testing

A large number of alleles from the six different short tandem repeat (STR) loci FGA, D3S1358, vWA, CSF1PO, TPOX and TH01, used in human identity testing were sequenced to provide support for the robustness of fluorescent STR DNA typing by allele size. Sequence information for some of these loci (FGA, vWA, TH01) is an extension of published work, whereas no extensive sequence information is available with respect to the D3S1358, CSF1PO, and TPOX loci. Sequencing of alleles at each locus has provided quantitative data with respect to the true nucleotide length of common alleles, and of alleles that vary in length from the common alleles. All alleles that were identified as "off-ladder" alleles through fluorescent typing at these STR loci have proven to be true length variant alleles. Sequencing at the D3S1358 and CSF1PO loci allowed for the establishment of a common nomenclature for these loci. A correlation between percent stutter and the length of the core tandem repeat is demonstrated at the FGA locus. Alleles in which the core tandem repeat is interrupted by a repeat unit of different sequence have a reduced percent stutter. DNA samples from three non-human primates (chimpanzee, orangutan, and gorilla) were compared to the human sequences, and shown to differ markedly across loci with respect to their homology. The effects of primer binding site mutations on the amplification efficiency at a particular locus, and methods used to interpret amplification imbalance of heterozygous alleles at a locus is also addressed.     

A LDR‐PCR Approach for Multiplex Polymorphisms Genotyping of Severely Degraded DNA with Fragment Sizes <100 bp*

Abstract: Reducing amplicon sizes has become a major strategy for analyzing degraded DNA typical of forensic samples. However, amplicon sizes in current mini‐short tandem repeat‐polymerase chain reaction (PCR) and mini‐sequencing assays are still not suitable for analysis of severely degraded DNA. In this study, we present a multiplex typing method that couples ligase detection reaction with PCR that can be used to identify single nucleotide polymorphisms and small‐scale insertion/deletions in a sample of severely fragmented DNA. This method adopts thermostable ligation for allele discrimination and subsequent PCR for signal enhancement. In this study, four polymorphic loci were used to assess the ability of this technique to discriminate alleles in an artificially degraded sample of DNA with fragment sizes <100 bp. Our results showed clear allelic discrimination of single or multiple loci, suggesting that this method might aid in the analysis of extremely degraded samples in which allelic drop out of larger fragments is observed.     

Sequence characterization of microvariant alleles at DYS627 and DYS458

Y chromosome short tandem repeats (Y-STRs) have been widely used in genetic applications and forensic casework. Recently, we found two intermediate alleles, the DYS627 allele 24.1 and the DYS458 allele 15.3, from Chinese Han population. The two allelic variants have not been recorded by the YHRD database. We have examined the molecular structure of these allelic variants by Sanger sequencing. The results showed that this intermediate allele at DYS627 was confirmed as 24.1, the sequence of which showed a base “A” insertion in the 13th repeat unit, and the intermediate allele at DYS458 was confirmed as 15.3, the sequence of which showed a base “G” deletion in the 12th repeat unit. This may be important for individual identification and paternal kinship testing. Besides, more allelic variants detected can be enriched in the Y-STR database.     

Distribution of HLA-DQA1 alleles in Arab and Pakistani individuals from Dubai,United Arab Emirates

PCR-based typing of the HLA-DQA1 locus, using allele specific oligonucleotide (ASO) probes and reverse dot blot methodology was used to determine allelic distributions and construct a database for Arab and Pakistani individuals living in Dubai. Genotype and allelic frequencies were calculated, and the data were tested for departures from Hardy-Weinberg (HWE) equilibrium. The most frequent HLA-DQA1 alleles among Dubaian Arabs are DQA1 4 and 1.2. Among Pakistanis, the most frequent allele is also DQA1 4. No significant deviations from HWE were detected.     

The different mobility of complementary strands depends on the proportion AC/GT

The electrophoretic mobility of DNA fragments on denaturing polyacrylamide gel depends on various factors. One of these is the base composition of a single-stranded DNA (ssDNA). We confirmed that one strand and its complementary strand of polymerase chain reaction (PCR) products migrated with different mobilities in all alleles detected at 12 out of the 13 short tandem repeat (STR) loci studied. The mobility differences between complementary strands (MD) were also observed regardless of end-polishing with Pfu DNA polymerase. MD was therefore not influenced by additional nucleotides to each strand of the PCR products. We then analyzed the relation between MD and the base composition using one representative allele at each of the 13 loci. The results indicated that MD was affected by the adenine plus cytosine (AC) content in the ssDNA and was proportional to the values of the AC content divided by the guanine plus thymine (GT) content in the AC-rich strand (the proportion AC/GT). When the proportion AC/GT was well-balanced, MD decreased. The same tendency was observed even in the end-polished strands. In this study, the electrophoretic mobility of an ssDNA on denaturing polyacrylamide gels was shown to depend on the proportion AC/GT. Unless the same side of the PCR products is labelled in the context of a PCR-based STR typing, distinct alleles may be mistaken for identical ones because of the different mobility of complementary strands. Accordingly, the labelled strand should be described if only one strand of the PCR products is detected. When using an allelic ladder marker as a size standard, the labelled side should be unified between STR alleles and the allelic ladder alleles.     

Further characterization and population data for the pentanucleotide STR polymorphism D10S2325.

Pentanucleotide tandem repeat markers are interesting for forensic sciences, because they may present less stutter on the electrophoretic pattern. We focused on the analysis of the DNA sequence for each allele at the pentanucleotide STR locus D10S2325 in order to understand their structures in the human genome and to construct human allelic ladder, which is necessary for forensic DNA typing. In order to evaluate the forensic applicability of D10S2325 and to construct a preliminary database, the genotype distributions and allele frequencies in three major ethnic groups were investigated. The population samples included Caucasians (Germans), Africans (African Americans), and Asians (Chinese). A total of 520 samples from unrelated individuals was analyzed by Amp-FLP. An example of each allele and new alleles were sequenced. Allele determination was carried out by comparison with a sequenced human allelic ladder made in-house. This pentanucleotide STR provided easily interpretable results. A total of 15 alleles was found in our population samples. Three new alleles were observed and named as alleles 19 and 21 based on the number of repeat motifs, while allele 19 can be divided further into two alleles, 19a and 19 according to analysis of the sequence. No evidence of deviation from Hardy-Weinberg equilibrium was observed. In 64 confirmed father/mother/child triplets no mutation event was observed. Using a maximum likelihood method, the mutation rate was indirectly estimated as 2.5 x 10(-5). These results suggest that D10S2325 is a useful marker for forensic casework and paternity analysis.     

Non-human Primate Studies Inform the Foundations of Fair and Just Human Institutions

Experiments with human participants have inspired new theories to capture human social, economic, and justice preferences, and shed new light on the foundation of institutions that promote and support large-scale exchange. Another source of valuable data for informing this agenda derives from studies with non-human primates. Here, we argue that primate studies of social preferences provide behavioral evidence supporting the role of the brain as an evolved social record-keeping device. Our argument follows Dickhaut et al. (Accounting Horizons 24:221?C255, 2010), who pointed to record-keeping as critical in enabling large-scale trade. Here, we note that record-keeping also underlies justice judgments in both personal exchange and large-scale trade. The reason is that evaluating whether an allocation is just requires tracking not only benefits that accrue locally, but also benefits for distant others. Further, if record-keeping is an evolved trait (as Dickhaut et al. in Accounting Horizons 24:221?C255, 2010 suggest), then it seems reasonable to expect it to be evidenced not only in humans, but also in non-human primates. Indeed, we argue that evidence from non-human primate research supports the Dickhaut hypothesis, thus supporting the role of justice in the emergence of fair and efficient economic exchange.     

The Influence of the Relationship and Motivation on Inequity Aversion in Dogs

One crucial element for the evolution of cooperation may be the sensitivity to others?? efforts and pay-offs in comparison with one??s own costs and gains. If an individual responds to a disadvantageous reward distribution, then it would likely increase its relative fitness compared with those who do not (Brosnan & de Waal, Nature 428:140, 2004). Recent experimental research indicates that sensitivity toward unequal reward distribution is not a uniquely human trait; non-human primates react to inequity if they witness a conspecific that obtains a more attractive reward for the same effort. However, primates are not the only species that cooperate and thus would benefit from inequity aversion. Canids also frequently cooperate with group members (e.g., wolves, wild dogs, etc.) or humans (domestic dogs), allowing for the possibility that they also are sensitive to the reward distribution. We report the findings of two studies of dogs?? responses to inequity and the social factors that mediate such responses. In the first study, we investigated whether domestic dogs showed a response to the inequity of rewards received for the same action in pairs of familiar dogs. We found that dogs showed significant behavioral differences when tested without food rewards when in the presence of a rewarded partner compared to the baseline and asocial control situations. This indicated that the presence of a rewarded partner matters (Range, Horn, Virányi, & Huber, Proc Natl Acad Sci USA 106(1):340?C345, 2009). In contrast to primates, dogs did not react to differences in the quality of food or effort. In the second study, we analyzed whether individual (motivational or personality) characteristics determined the response of each subject to unequal reward distributions or whether the subject??s responses depended on the specific relationship they had with its partner. We found that individual motivation and the quality of the relationship influenced the dogs?? performance in test conditions, but that these factors varied across the control and reward inequity conditions. Overall, our results suggest that inequity aversion in dogs is conditional on their and their partner??s rewards. Whether inequity aversion is based on the same mechanisms in both humans and non-human primates is still unclear.     

Cross-Disciplinary Conversations: A Psychological Perspective on Justice Research with Non-human Animals

Social Justice Research has devoted two recent issues to the topic of inequity responses in non-human animals. The goal of this paper is to provide some commentary from the perspective of psychological theory and research on justice and fairness in humans. In an attempt to build greater cross-disciplinary sharing of ideas and insights, I briefly review the major insights from (a) contemporary research on questions of fairness and justice with non-human primates and how this corpus of knowledge can inform the on-going study of these issues in psychology and related disciplines, and (b) 50-plus years of research on justice and fairness in psychology and related disciplines, and how it can inform contemporary research efforts with non-human animals going forward. The spotlight behavioral economists and justice research with non-human animals places on the primary role of distributive justice is suggestive that it may be time for a renaissance of interest in this topic in psychology and related disciplines. The focus of psychological research on topics such as boundary conditions on equity as key justice concern (e.g., alternative distributive norms such as equality and need), as a well as attention given to procedural, interactional, retributive, and restorative justice, is suggestive that research with non-human animals should broaden its horizons to study alternative conceptions of justice and fairness.     

Genetic analysis of fingerprints—Could WGA or nested-PCR be alternatives to the increase of PCR cycles number?

In this work we aimed to compare the application of increasing PCR cycle number, whole genome amplification and nested-PCR on fingerprints genetic analysis. Results were compared for correct alleles, allele dropin and allelic dropout. We concluded that increasing the number of PCR cycles is yet the best way to attain the required sensitivity.