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1.
Zhongguo Zhong Yao Za Zhi ; 45(15): 3659-3665, 2020 Aug.
Artigo em Chinês | MEDLINE | ID: mdl-32893555

RESUMO

As a traditional Chinese medicinal material, Chrysosplenium is urgently needed for genetic resource investigation and protection research due to the decrease of its wild resources in recent years. After investigating the wild resources, we conducted genetic polymorphism and clustering studies of 24 species(a total of 36 samples) of Chrysosplenium using SRAP technique. The results showed that a total of 374 polymorphic bands were obtained using 18 pairs of SRAP primers to amplify these samples, on average of 20.7 bands for each primer pair. We used the biological software to analyze the population's genetic parameter and got the N_a value as 2.000 0, N_(e )value as 1.408 4, the average Nei's index as 0.263 5, and the average Shannon information index as 0.419 1. UPGMA cluster analysis showed that all the samples can be divided into three major groups at the genetic similarity coefficient of 0.70: there are 18 species(24 samples) gathered for the Ⅰ groups, 3 species or variation(7 samples) for Ⅱ groups, and 3 species(5 samples) for Ⅲ groups. The differences of these Chrysosplenium species at the molecular level are consistent with that of their geographical and ecological distribution. At the same time, we used SRAP technology to construct 36 DNA digital fingerprints of Chrysosplenium and obtained the unique molecular identification band type of each material. These results will provide effective methods and reliable basis for the identification, protection and genetic diversity analysis of the germplasm resources of Chrysosplenium, and lay a foundation for the further development and utilization of them.


Assuntos
Impressões Digitais de DNA , Variação Genética , Análise por Conglomerados , Marcadores Genéticos , Filogenia , Polimorfismo Genético
2.
PLoS One ; 15(7): e0235484, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-32634144

RESUMO

Accurate identification of crop varieties grown by farmers is crucial, among others, for crop management, food security and varietal development and dissemination purposes. One may expect varietal identification to be more challenging in the context of developing countries where literacy and education are limited and informal seed systems and seed recycling are common. This paper evaluates the extent to which smallholder farmers misidentify their wheat varieties in Ethiopia and explores the associated factors and their implications. The study uses data from a nationally representative wheat growing sample household survey and DNA fingerprinting of seed samples from 3,884 wheat plots in major wheat growing zones of Ethiopia. 28-34% of the farmers correctly identified their wheat varieties. Correct identification was positively associated with farmer education and seed purchases from trusted sources (cooperatives or known farmers) and negatively associated with seed recycling. Farmers' varietal identification thereby is problematic and leads to erroneous results in adoption and impact assessments. DNA fingerprinting can enhance varietal identification but remains mute in the identification of contextual and explanatory factors. Thus, combining household survey and DNA fingerprinting approaches is needed for reliable varietal adoption and impact assessments, and generate useful knowledge to inform policy recommendations related to varietal replacement and seed systems development.


Assuntos
Produtos Agrícolas/genética , Impressões Digitais de DNA , Sementes/genética , Triticum/genética , Agricultura , Produtos Agrícolas/classificação , Produtos Agrícolas/crescimento & desenvolvimento , Etiópia , Fazendeiros , Humanos , Sementes/classificação , Sementes/crescimento & desenvolvimento , Triticum/classificação , Triticum/crescimento & desenvolvimento
3.
Forensic Sci Rev ; 32(2): 105-116, 2020 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-32712578

RESUMO

The challenge of profiling spermatozoa from samples containing a mixture of male and female cells has been extensively discussed within the forensic community. Various techniques have been developed for the analysis of sexual assault evidence with the aim to generate a single-source male DNA profile. Multiple methods practiced for the isolation of the male component are discussed in this review, with a focus on differential extraction. Benefits of alterations that have been made to the original differential method to increase the efficiency are highlighted. Although improvements were achieved, it is ascertained by this review that these methods are limited in their overall success rate or their applicability. Perhaps future approaches and research should concentrate on more efficient, cost-effective, and time-saving techniques to individually sort or isolate spermatozoa.


Assuntos
Ciências Forenses , Delitos Sexuais , Espermatozoides , Impressões Digitais de DNA , Feminino , Ciências Forenses/métodos , Humanos , Masculino
5.
Leg Med (Tokyo) ; 46: 101719, 2020 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-32512465

RESUMO

In order to create an autosomal STR loci population database for Himachal Pradesh, 259 blood samples were taken from people residing in various regions of the state and AmpFlSTR® Identifiler® Plus PCR amplification kit was used for evaluation of 15 autosomal STR markers. A total of 149 alleles were investigated in this study with a mean allele number of 9.933 per locus. The locus D2S1338 was most informative in our data, as it had the highest discrimination power (PD-0.967) and the highest polymorphic information content (PIC-0.86). The matching probability and typical paternity index for all the studied loci were observed as 2.9x10-18 and 4.7x105, respectively. Discrimination power (CPD) and exclusion power (CPE) for all the studied loci were observed as 1 and 0.999998.


Assuntos
Impressões Digitais de DNA , Bases de Dados Genéticas , Genética Forense/métodos , Loci Gênicos/genética , Repetições de Microssatélites/genética , Alelos , Genômica , Humanos , Índia , Reação em Cadeia da Polimerase
6.
Leg Med (Tokyo) ; 46: 101713, 2020 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-32442862

RESUMO

An evaluation of a Rapid DNA system was performed using buccal swab samples and mock Disaster Victim Identification (DVI) samples collected postmortem. The allelic ladder success rate was 90% and samples analyzed simultaneously with this allelic ladder were used for further analysis. Sample success rate of the Rapid DNA system for buccal swab samples, and blood and muscle DVI samples were calculated. Success rates of buccal swab samples were 100% and 75% using cassettes preloaded with all reagents suitable for high- and low-DNA content samples, respectively. Success rates of fresh DVI samples were 80% to 100%. Success rates of putrefied DVI samples varied widely between 0% and 20% and 50% to 80% depending on cassette and sample types. Conventional DNA analysis was performed for comparison with the results of the Rapid DNA system. DNA quantity and degradation of human DNA were measured using quantitative polymerase chain reaction. DVI samples that yielded more than 1 ng/µL of DNA when extracted with conventional protocols were suitable for analysis using cassettes for both high- and low-DNA content samples. DVI samples with less than 0.1 ng/µL of DNA were suitable only for analysis using cassettes for low-DNA content samples. All alleles called and exported by the Expert system software implemented in the Rapid DNA system were concordant with allele calls made by conventional capillary electrophoresis DNA analysis.


Assuntos
Impressões Digitais de DNA/métodos , Vítimas de Desastres , Medicina Legal/métodos , Mucosa Bucal , DNA/análise , Humanos , Manejo de Espécimes
7.
Gene ; 753: 144794, 2020 Aug 30.
Artigo em Inglês | MEDLINE | ID: mdl-32464245

RESUMO

Novel microsatellite markers were developed to investigate the genetic diversity and DNA fingerprinting of bougainvillea cultivars. Total of 175 SSRs were designed from over 50,000 SSRs identified in the whole genome sequence data, 33 highly polymorphic markers were identified. These selected SSRs produced a total of 165 alleles with 2 (BOUG-3 and BOUG-50) to 9 (BOUG-69) alleles per loci with an average of 5 alleles per locus. The overall size of the amplified products ranged from 90 bp (BOUG-51 and BOUG-81) to 320 bp (BOUG-162). The gene diversity per locus ranged from 0.13 to 0.91 with a mean of 0.71. Primer BOUG-73 and BOUG-124 exhibited highest gene diversity with greater number of alleles. The mean Nei's genetic diversity index was 0.678 with range of 0.134 (BOUG-77) to 0.958 (BOUG-69). The UPGMA based dendrogram divided the cultivars into seven major clusters. Clustering pattern was more distinct for bract types and variegated cultivars which were also confirmed by PCA scatter plot diagram. The pair-wise genetic distance estimates ranged from 0.089 to 0.86 with an average of 0.56. Each of the 125 cultivar profiled had unique marker profile indicating that the SSR markers identified are useful for identification and differentiation of bougainvillea cultivars. These informative markers identified from the study will be of great utility to assess the genetic diversity, understanding the population structure and in marker assisted breeding for improvement of bougainvillea.


Assuntos
Repetições de Microssatélites/genética , Nyctaginaceae/genética , Alelos , Impressões Digitais de DNA/métodos , Marcadores Genéticos/genética , Variação Genética , Genótipo , Filogenia , Melhoramento Vegetal/métodos , Polimorfismo Genético
8.
Zhongguo Zhong Yao Za Zhi ; 45(5): 1064-1069, 2020 Mar.
Artigo em Chinês | MEDLINE | ID: mdl-32237447

RESUMO

The pig bile powder, bovine bile powder, snake bile, sheep bile, goose bile powder, and bear bile powder were contained by the Chinese Pharmacopoeia. The bile power medicine has a long history in traditional Chinese medicine and definite effect. However, the medicine of bile powder(bile) are similar in morphology. Besides, many medicine lack specific microscopic identification characteristics and chemical characteristics. There is a risk of adulteration, especially when the fake medicine were mixed in authentic medicine, it is difficult to detection. The key to control the quality and ensures the clinical efficacy is the good or bad, true or false of the bile power medicine. The STR typing technology is a method that according to differential typing of PCR amplified lengths to compare and identify individual organisms. Based on the principle of STR typing, the easily, rapid DNA fingerprinting method to identify the bile power and adulteration was established.The original animal or bile powder of pigs, cattle, sheep, chickens, ducks, geese, snakes, bears, fish were collected, the 12 S-L1091/12 S-H1478 and 16 S-L3428/16 S-H3667 was obtained by sifted, the DNA fingerprinting of the bile power and adulteration was obtained by STR typing. Every species has different STR fingerprints, so different species can be identified. Besides, the fingerprints have both the authentic and fake's information, the adulteration of authentic and fake can be identified. Therefore, the method to identify the bile power and adulteration was achieved through the combination of two primers. The DNA fingerprinting method established in this study can also be used for other animal medicine.


Assuntos
Bile/química , Impressões Digitais de DNA , Materia Medica/análise , Animais , Bovinos , Galinhas , Medicina Tradicional Chinesa , Ovinos , Suínos , Ursidae
9.
Biol Res ; 53(1): 13, 2020 Apr 15.
Artigo em Inglês | MEDLINE | ID: mdl-32293552

RESUMO

BACKGROUND: Gallbladder cancer (GBC) is the most common tumor of the biliary tract. The incidence of GBC shows a large geographic variability, being particularly frequent in Native American populations. In Chile, GBC represents the second cause of cancer-related death among women. We describe here the establishment of three novel cell lines derived from the ascitic fluid of a Chilean GBC patient, who presented 46% European, 36% Mapuche, 12% Aymara and 6% African ancestry. RESULTS: After immunocytochemical staining of the primary cell culture, we isolated and comprehensively characterized three independent clones (PUC-GBC1, PUC-GBC2 and PUC-GBC3) by short tandem repeat DNA profiling and RNA sequencing as well as karyotype, doubling time, chemosensitivity, in vitro migration capability and in vivo tumorigenicity assay. Primary culture cells showed high expression of CK7, CK19, CA 19-9, MUC1 and MUC16, and negative expression of mesothelial markers. The three isolated clones displayed an epithelial phenotype and an abnormal structure and number of chromosomes. RNA sequencing confirmed the increased expression of cytokeratin and mucin genes, and also of TP53 and ERBB2 with some differences among the three cells lines, and revealed a novel exonic mutation in NF1. The PUC-GBC3 clone was the most aggressive according to histopathological features and the tumorigenic capacity in NSG mice. CONCLUSIONS: The first cell lines established from a Chilean GBC patient represent a new model for studying GBC in patients of Native American descent.


Assuntos
Antígenos Glicosídicos Associados a Tumores/genética , Neoplasias da Vesícula Biliar/genética , Índios Sul-Americanos/genética , Animais , Antineoplásicos/farmacologia , Líquido Ascítico/metabolismo , Carcinogênese/genética , Testes de Carcinogenicidade , Linhagem Celular Tumoral/efeitos dos fármacos , Linhagem Celular Tumoral/metabolismo , Chile , Cisplatino/farmacologia , Células Clonais/efeitos dos fármacos , Células Clonais/metabolismo , Impressões Digitais de DNA , Desoxicitidina/análogos & derivados , Desoxicitidina/farmacologia , Células Epiteliais/metabolismo , Neoplasias da Vesícula Biliar/metabolismo , Perfilação da Expressão Gênica , Genes erbB-2/genética , Humanos , Queratina-19/genética , Queratina-7/genética , Masculino , Camundongos Endogâmicos NOD , Pessoa de Meia-Idade , Receptor ErbB-2/genética , Análise de Sequência de RNA , Células Tumorais Cultivadas , Proteína Supressora de Tumor p53/genética
10.
J Water Health ; 18(1): 19-29, 2020 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-32129183

RESUMO

Fecal contamination is one of the factors causing deterioration of Laguna Lake. Although total coliform levels are constantly monitored, no protocol is in place to identify their origin. This can be addressed using the library-dependent microbial source tracking (MST) method, repetitive element sequence-based polymerase chain reaction (rep-PCR) fingerprinting. Serving as a prerequisite in developing the host-origin library, we assessed the discriminatory power of three fingerprinting primers, namely BOX-A1R, (GTG)5, and REP1R-1/2-1. Fingerprint profiles were obtained from 290 thermotolerant Escherichia coli isolated from sewage waters and fecal samples of cows, chickens, and pigs from regions surrounding the lake. Band patterns were converted into binary profiles and were classified using the discriminant analysis of principal components. Results show that: (1) REP1R-1/2-1 has a low genotyping success rate and information content; (2) increasing the library size led to more precise estimates of library accuracy; and (3) combining fingerprint profiles from BOX-A1R and (GTG)5 revealed the best discrimination (average rate of correct classification (ARCC) = 0.82 ± 0.06) in a two-way categorical split; while (4) no significant difference was found between the combined profiles (0.74 ± 0.15) and using solely BOX-A1R (0.76 ± 0.09) in a four-way split. Testing the library by identifying known isolates from a separate dataset has shown that a two-way classification performed better (ARCC = 0.66) than a four-way split (ARCC = 0.29). The library can be developed further by adding more representative isolates per host source. Nevertheless, our results have shown that combining profiles from BOX-A1R and (GTG)5 is recommended in developing the MST library for Laguna Lake.


Assuntos
Impressões Digitais de DNA , Monitoramento Ambiental/métodos , Lagos/microbiologia , Poluentes da Água/análise , Animais , Bovinos , Galinhas , Fezes/microbiologia , Feminino , Filipinas , Reação em Cadeia da Polimerase , Suínos
11.
PLoS Negl Trop Dis ; 14(3): e0008077, 2020 03.
Artigo em Inglês | MEDLINE | ID: mdl-32214313

RESUMO

BACKGROUND: Phlebotomus (Larroussius) perniciosus and Canis familiaris are respectively the only confirmed vector and reservoir for the transmission of Leishmania (L.) infantum MON-1 in Tunisia. However, the vector and reservoir hosts of the two other zymodemes, MON-24 and MON-80, are still unknown. The aim of this study was to analyze the L. infantum life cycle in a Tunisian leishmaniasis focus. For this purpose, we have focused on: i) the detection, quantification and identification of Leishmania among this sand fly population, and ii) the analysis of the blood meal preferences of Larroussius (Lar.) subgenus sand flies to identify the potential reservoirs. METHODOLOGY AND FINDINGS: A total of 3,831 sand flies were collected in seven locations from the center of Tunisia affected by human visceral leishmaniasis. The collected sand flies belonged to two genus Phlebotomus (Ph.) (five species) and Sergentomyia (four species). From the collected 1,029 Lar. subgenus female sand flies, 8.26% was positive to Leishmania by ITS1 nested PCR. Three Leishmania spp. were identified: L. infantum 28% (24/85), L. killicki 13% (11/85), and L. major 22% (19/85). To identify the blood meal sources in Ph. Lar. subgenus sand flies, engorged females were analyzed by PCR-sequencing targeting the vertebrate cytochrome b gene. Among the 177 analyzed blood-fed females, 169 samples were positive. Sequencing results showed seven blood sources: cattle, human, sheep, chicken, goat, donkey, and turkey. In addition, mixed blood meals were detected in twelve cases. Leishmania DNA was found in 21 engorged females, with a wide range of blood meal sources: cattle, chicken, goat, chicken/cattle, chicken/sheep, chicken/turkey and human/cattle. The parasite load was quantified in fed and unfed infected sand flies using a real time PCR targeting kinetoplast DNA. The average parasite load was 1,174 parasites/reaction and 90 parasites/reaction in unfed and fed flies, respectively. CONCLUSION: Our results support the role of Ph. longicuspis, Ph. perfiliewi, and Ph. perniciosus in L. infantum transmission. Furthermore, these species could be involved in L. major and L. killicki life cycles. The combination of the parasite detection and the blood meal analysis in this study highlights the incrimination of the identified vertebrate in Leishmania transmission. In addition, we quantify for the first time the parasite load in naturally infected sand flies caught in Tunisia. These findings are relevant for a better understanding of L. infantum transmission cycle in the country. Further investigations and control measures are needed to manage L. infantum transmission and its spreading.


Assuntos
DNA/análise , Comportamento Alimentar , Conteúdo Gastrointestinal/química , Conteúdo Gastrointestinal/parasitologia , Especificidade de Hospedeiro , Leishmania infantum/isolamento & purificação , Phlebotomus/fisiologia , Animais , DNA/genética , Impressões Digitais de DNA , DNA Espaçador Ribossômico/genética , Transmissão de Doença Infecciosa , Feminino , Humanos , Leishmania infantum/genética , Masculino , Phlebotomus/parasitologia , Reação em Cadeia da Polimerase , Análise de Sequência de DNA , Tunísia
12.
Anthropol Anz ; 77(3): 235-242, 2020 Aug 12.
Artigo em Inglês | MEDLINE | ID: mdl-32211746

RESUMO

This paper discusses our approach and results obtained when attempting to identify a saponified human body recovered from the sea, without arms and legs. Bones, especially the long ones, are the only sources of DNA available in several cases involving unidentified bodies in advanced state of putrefaction. In this case, since the body was found without limbs, attempts were made to extract DNA from the sternum bone. The DNA was extracted using a modified version of the NucleoSpin® DNA Trace Kit (Macherey Nagel™) protocol and an STR analysis was performed. Thanks to this modified protocol a complete DNA profile was obtained from the sternum bone, while only partial results were obtained from blood and teeth. The DNA profile obtained from the sternum was compared with the DNA of the putative son searching for a genetic match. Five incompatibilities were detected so it was possible to exclude the kinship. In conclusion this could be a useful technique in personal identification through DNA analysis in case of poor quality and quantity of bone.


Assuntos
Impressões Digitais de DNA , DNA , Dente , Antropologia Física , DNA/isolamento & purificação , Humanos , Repetições de Microssatélites , Esterno
13.
PLoS One ; 15(3): e0230390, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-32176736

RESUMO

The aim of the study was to detect and genetically characterize Arcobacter butzleri in pet red-footed tortoises suspected for Campylobacter spp., using molecular techniques. A written consent from tortoise owners was obtained, after explaining the advantages of the research to tortoise owners of Grenada. Fecal samples were collected from 114 tortoises from five parishes of the country and cultured for Campylobacter spp. using selective culture techniques. A. butzleri was isolated from 4.39% of pet tortoises. Total thirteen isolates were obtained; all identified as A. butzleri by a universal and a species-specific Polymerase Chain Reaction (PCR) and direct sequencing. Genetic characterization of these isolates was performed based on Enterobacterial repetitive intergenic consensus PCR (ERIC-PCR) that generated eight different genetic fingerprints with a discriminatory power of 0.91. Campylobacter species were not detected molecularly in any of the culture-positive samples. This is the first report of infection of pet tortoises in Grenada, West Indies with A. butzleri. This study emphasizes on the risk of zoonotic transmission of A. butzleri by exotic pets, which is a serious concern for public health.


Assuntos
Arcobacter/genética , Campylobacter/genética , Sequências Repetitivas de Ácido Nucleico/genética , Tartarugas/microbiologia , Animais , Campylobacter/isolamento & purificação , Impressões Digitais de DNA/métodos , DNA Bacteriano/genética , DNA Bacteriano/isolamento & purificação , Fezes/microbiologia , Tartarugas/genética
14.
Nat Neurosci ; 23(3): 375-385, 2020 03.
Artigo em Inglês | MEDLINE | ID: mdl-32015540

RESUMO

Autism spectrum disorder (ASD) is genetically heterogeneous with convergent symptomatology, suggesting common dysregulated pathways. In this study, we analyzed brain transcriptional changes in five mouse models of Pitt-Hopkins syndrome (PTHS), a syndromic form of ASD caused by mutations in the TCF4 gene, but not the TCF7L2 gene. Analyses of differentially expressed genes (DEGs) highlighted oligodendrocyte (OL) dysregulation, which we confirmed in two additional mouse models of syndromic ASD (Ptenm3m4/m3m4 and Mecp2tm1.1Bird). The PTHS mouse models showed cell-autonomous reductions in OL numbers and myelination, functionally confirming OL transcriptional signatures. We also integrated PTHS mouse model DEGs with human idiopathic ASD postmortem brain RNA-sequencing data and found significant enrichment of overlapping DEGs and common myelination-associated pathways. Notably, DEGs from syndromic ASD mouse models and reduced deconvoluted OL numbers distinguished human idiopathic ASD cases from controls across three postmortem brain data sets. These results implicate disruptions in OL biology as a cellular mechanism in ASD pathology.


Assuntos
Transtorno do Espectro Autista/genética , Impressões Digitais de DNA , Hiperventilação/genética , Deficiência Intelectual/genética , Bainha de Mielina/genética , Transcriptoma/genética , Envelhecimento , Animais , Contagem de Células , Facies , Regulação da Expressão Gênica , Humanos , Proteína 2 de Ligação a Metil-CpG/genética , Camundongos , Camundongos Knockout , Oligodendroglia/metabolismo , PTEN Fosfo-Hidrolase/genética , Cultura Primária de Células , Transdução de Sinais/genética , Fator de Transcrição 4/genética
15.
Environ Sci Pollut Res Int ; 27(7): 7639-7646, 2020 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-31889276

RESUMO

Like other vegetables, Pisum sativum L. also faces storage and degradation problems. To enhance their resistance and make them enable to cope with the deterioration problems during storage, the current study was designed to develop two resistant lines of P. sativum in terms of phenolic contents and genotypes. The phenolic compounds generally have antioxidant properties and deterioration during storage which are usually due to oxidation caused by free radicals. Thus, if a variety has high phenolic contents these problems will be coped in a better way. The genotype of a plant is also important in this regard, and the best adopted species would survive in unfavorable conditions. First, the phenolic and flavonoid contents were determined in the crude extract using the Folin-Ciocalteu method. Then, the identification and quantification of phenolic compounds were carried out in the developed lines of selected plants PL-04 and PL-05, as well as in the parental varieties [Climax (female) and Falan (male)] via HPLC. DPPH assay was used to determine the free radical scavenging capabilities of the extracts of the developed verities. The genotypic differences were confirmed by DNA fingerprinting using advanced simple sequence repeat (SSR) markers. The HPLC analysis of PL-04 confirmed the presence of three phenolic compounds in an appreciable amount which exhibited a higher antioxidant activity against DPPH radicals, while in the parental varieties, two phenolic compounds were identified and exhibited lower antioxidant activities. PL-04 was found rich in phenolic compounds and affectively scavenge-free radicals which would therefore be resistant to oxidation and degradation caused by free radicals. Comparing the present findings with our previous one, P-04 was found to be resistant to powdery mildew; it was concluded that the most probable reason of the resistance was the high phenolic contents and thus long shelf life.


Assuntos
Antioxidantes/química , Ervilhas/química , Fenóis/química , Impressões Digitais de DNA , Flavonoides/química , Repetições de Microssatélites , Oxirredução , Extratos Vegetais/química
16.
Forensic Sci Int ; 307: 110114, 2020 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-31901461

RESUMO

Seminal evidence obtained from a sexual crime scene provides clues for solving a case through forensic analysis. However, most evidence collected from sexual crime scenes is a mixture of sperm cells and vaginal discharge. Therefore, separating the sperm cells from the seminal evidence is very important. In this study, we developed a separation method for effectively separating sperm cells using differential extraction with commercially available sperm staining reagents such as hematoxylin and nigrosin. Hematoxylin (0.03 % v/v) effectively stained the sperm cells in ATL and TNE lysis buffer, while nigrosin did not. The loss of sperm cells during washing of the specimen was minimized using the differential extraction method. Subsequently, genomic DNA was extracted from the hematoxylin-stained sperm cells and subjected to short tandem repeat genotyping. We observed no interference from hematoxylin. These results indicate that hematoxylin can be used to stain sperm cells and thus facilitate subsequent genetic identification.


Assuntos
Separação Celular , Hematoxilina , Delitos Sexuais , Espermatozoides/química , Espermatozoides/citologia , Compostos de Anilina , DNA/isolamento & purificação , Impressões Digitais de DNA , Eletroforese Capilar , Feminino , Genética Forense , Técnicas de Genotipagem , Humanos , Indicadores e Reagentes , Masculino , Repetições de Microssatélites , Reação em Cadeia da Polimerase , Coloração e Rotulagem
17.
Forensic Sci Int ; 307: 110142, 2020 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-31927396

RESUMO

Forensic science is one of the most modern and applied fields of science, today and comprises of various domains. These include Fingerprints analysis, Questioned document analysis, Forensic DNA and serology, Anthropometry, Cyber and Digital forensics, and many other fields. All these fields aid the process of decision making in the courts of law and legal settings; however, DNA profiling and its analyses are one of the most important aspects of forensic science today. In Forensic DNA analysis, the statistical calculations are very important to estimate the conclusiveness of DNA evidence in forensic cases; and to establish paternity and relatedness in civil and criminal matters. These statistics, when performed manually, leave a chance of error or ambiguity in the calculation, and are hectic and time-taking. Therefore, the computer-aided approaches are opted in forensics to perform DNA statistics calculations. Keeping its importance in mind, a highly accurate windows-based software program namely ForeStatistics is proposed in this study. ForeStatistics is rich in features such as DNA statistical calculations, DNA profile management and its matching. The software can estimate random match probabilities for single-source profiles, combined probability of inclusion for mixed profiles, paternity index of a disputed child in duo and trio cases, paternity of the disputed child when the alleged father is related to mother or biological father and relatedness in cases of grandparents/grandchild, avuncular relation and cousin. It is validated through different protocols and the validation of ForeStatistics depicts that it is highly accurate in terms of performing DNA statistics or DNA profile matching. Thus, it is concluded, that ForeStatistics has a great utility in the field of Forensic DNA analysis and can help DNA scientists, in performing various DNA related statistics, accurately and very efficiently. ForeStatistics can be downloaded freely from (http://zeetu.org/forestatistics.html).


Assuntos
Biometria , Impressões Digitais de DNA , Paternidade , Software , Humanos , Funções Verossimilhança , Probabilidade , Interface Usuário-Computador
18.
Forensic Sci Int ; 307: 110139, 2020 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-31935660

RESUMO

Cars are often sampled for DNA to help identify occupants and their possible location(s) within the car. While DNA from the frequent driver is likely to accumulate over time, DNA from previous and/or subsequent occupants, and those whose DNA has inadvertently been transferred to the car, may also contribute to any samples collected. This study investigates how much DNA resides on various sites within cars, and who might contribute to these samples. A total of 35-36 sites, internal and external, were targeted within four cars with sole long-term drivers. In addition to the car keys, sample sites included the exterior and interior door handles (driver and passenger sides), through to the internal compartments (driver side, middle area and front passengers' side). Reference samples were collected from the exclusive drivers, their co-resident partners and, where possible, recent passengers. The driver was always observed as a contributor in DNA profiles from the driver's side and, in most instances, was the sole, major or majority contributor to the profile. The driver was also observed as a major, majority or minor contributor at several sites on the passenger side. DNA of known recent passengers, close associates of the driver and unknown individuals was collected from many sites on both the driver and passenger sides. These findings may assist in sample targeting within cars and in the evaluation of DNA evidence when propositions relate to the activities performed.


Assuntos
Automóveis , Impressões Digitais de DNA , DNA/análise , Feminino , Genética Forense , Humanos , Funções Verossimilhança , Masculino , Repetições de Microssatélites , Reação em Cadeia da Polimerase , Prevalência , Manejo de Espécimes , Tato
19.
Am J Forensic Med Pathol ; 41(1): 56-59, 2020 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-31929321

RESUMO

Postmortem personal identification in forensic science is performed using various methods. However, severely burnt bodies are hard to identify using odontological or skeletal features because of carbonization, and sometimes DNA profiling is impracticable because of the unavailability of the relatives. We present a case of a burn victim found after a house fire. Personal identification was attempted, but the body was heavily charred to the bones and the use of physical appearance was impracticable. There were no known relatives or personal belongings of the deceased for comparison of DNA typing. We obtained a series of abdominal computed tomography (CT) scans taken antemortem and found bilateral multiple renal cysts, left renal artery calcification, and a big right inguinal hernia, which matched the deceased's postmortem CT findings and autopsy findings. To date, studies of identification by CT have acted for a rise in precision, but they require complicated calculation or high graphical methods. Calcification of the arteries or renal cysts seen in our case are very common lesions present in many adults with abundant variation; thus, they may be helpful as simple indicators for identification.


Assuntos
Fogo , Hérnia Inguinal/diagnóstico por imagem , Doenças Renais Císticas/diagnóstico por imagem , Artéria Renal/diagnóstico por imagem , Calcificação Vascular/diagnóstico por imagem , Autopsia/métodos , Queimaduras/patologia , Impressões Digitais de DNA , Medicina Legal/métodos , Humanos , Masculino , Pessoa de Meia-Idade , Tomografia Computadorizada por Raios X
20.
Sci Justice ; 60(1): 1-8, 2020 01.
Artigo em Inglês | MEDLINE | ID: mdl-31924284

RESUMO

Human biological samples with multiple contributors remain one of the most challenging aspects of DNA typing within a forensic science context. With the increasing sensitivity of commercially available kits allowing detection of low template DNA, complex mixtures are now a standard component of forensic DNA evidence. Over the years, various methods and techniques have been developed to try to resolve the issue of mixed profiles. However, forensic DNA analysis has relied on the same markers to generate DNA profiles for the past 30 years causing considerable challenges in the deconvolution of complex mixed samples. The future of resolving complicated DNA mixtures may rely on utilising markers that have been previously applied to gene typing of non-forensic relevance. With Massively Parallel Sequencing (MPS), techniques becoming more popular and accessible even epigenetic markers have become a source of interest for forensic scientists. The aim of this review is to consider the potential of alleles from the Human Leukocyte Antigen (HLA) complex as effective forensic markers. While Massively Parallel Sequencing of HLA is routinely used in clinical laboratories in fields such as transplantation, pharmacology or population studies, there have not been any studies testing its suitability for forensic casework samples.


Assuntos
Alelos , Impressões Digitais de DNA/métodos , Genética Forense/métodos , Marcadores Genéticos , Antígenos HLA/genética , Sequenciamento de Nucleotídeos em Larga Escala , Humanos , Repetições de Microssatélites , Polimorfismo de Nucleotídeo Único , Análise de Sequência de DNA
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