Detection of Deletion/Insertion Polymorphism Profiles from Single Human Hair Shafts

Background: Hair is a frequently encountered biological evidence in personal identification. The amount of nuclear DNA that can be extracted from a single strand of rootless hair is most limited, making the detection of short tandem repeat (STR) polymorphisms difficult. To overcome these limitations, deletion/insertion polymorphisms (DIP) as a new type of genetic marker have shown their benefits in detecting low-copy-number DNA. The Investigator DIPplex kit contains 30 biallelic autosomal DIP and amelogenin. The analysis of DIPs combines the advantages of both STR and single nucleotide polymorphism analyses. Thus, this study aimed to detect the DIP distribution of individual hair shafts from individuals.Methods and Results: DNA was extracted from the shaft of fresh, aged, and shed hair. After DNA was evaluated, the DIP profiles were detected by capillary electrophoresis. The results indicated that the amount of DNA extracted from hair roots was much higher than that from the hair shafts in the same individual for all samples. The degradation index values of DNA from the aged hair shafts were highest. It is classified to be “mildly degraded.” Compared with their hair roots, the full DIP profiles were detected for fresh hair, 70% for aged hair, and 92% for shed hair. Contrarily, except for fresh hair shafts, only three STR loci of the aged and shed strands of hair could be amplified using AmpFlSTR MiniFiler PCR Amplification Kit.Conclusions: These results indicate that the detection of DIP profile is an effective tool for personal identification from hair shafts, including aged hair.

FACILE IN VITRO GLASS SLIDE LIGHT MICROSCOPY METHOD USING TETRACYCLINE TO VISUALIZE A REPETITIVE PATTERN IN AERIAL PLANT ROOT TIPS FILAMENTOUS NETWORK

The main purpose of this manuscript is to introduce a facile light microscopy methodology to visualize plant roots filaments. In a previous manuscript in vitro experiments on freshly plucked human hair roots documented the commonly used antibiotic Tetracycline (TE) deleterious effect on soft tissue, severe enough to allow for visualization of an underlying filamentous skeleton. In this manuscript, TE was also evaluated in a similar fashion of in vitro experiments, this time aerial plant roots were immersed in liquid Tetracycline. Images and video recordings are presented where plant aerial root tissue cells appeared to interact with Tetracycline, thus allowing for exposure of an underlying filamentous network. These filaments were documented undergoing biosorption of Tetracycline, thus indicating a probable cellulose base. It is emphasized that a literature search showed similar, albeit visually different displays of roots filaments obtained by using a Scanning Electron Microscopy. The method herein introduced could be an adjunct to existing established methodology in root function research. Two salient advantages are identified, firstly that the essential minimal material and equipment is limited to a light microscope, glass slides, chosen biological material, water and powder Tetracycline. Secondly, the speed in obtaining results would offer researchers a preliminary or perhaps a final correct conclusion.

Estrus Behaviour and Reproductive Traits of Pulawska Gilts Associated with Selected Gene Polymorphisms

Searching for the associations between the gene polymorphism and the reproductive traits is essential in defining the genetic native breed specificity, which distinguishes them from the other breeds. The aim of our study was to determine the associations between mutations in the PRL, PRLR, PTGS2, FUT1 genes and sexual and periparturient activity in native Pulawska gilts. The analysis included 72 animals which gave birth to the first litter. Evaluation of the productive value of gilts accounted for indicators of sexual and periparturient activity as well as reproductive traits. The biological material for molecular analyses was obtained from the hair roots of the gilts. The genotype was verified by PCR RFLP analysis. The primers and PCR conditions were determined on the basis of available literature data. Statistically significant differences (P≤0.05) were found at the PRL locus: gilts of AA genotypes (Ins/Ins) at the PRL locus were characterised by longest farrowing duration compared to gilts of AB genotype (P≤0.05). The analysis of PRLR gene showed that gilts of TT genotype revealed a tendency for later occurrence of estrus signs (first and second estrus) and for the markedly longest farrowings (P≤0.05). With regard to PTGS2 and FUT1 loci, no significant differences were found in the parameters of sexual and periparturient activity of the gilts. However, gilts of FUT1 GG genotype gave birth to and reared the largest first litters (P≤0.05). The results of the studies expand the knowledge about the genetic structure and productivity of Pulawska gilts.

Smart Root Search (SRS): A Novel Nature-Inspired Search Algorithm

In this paper, a novel heuristic search algorithm called Smart Root Search (SRS) is proposed. SRS employs intelligent foraging behavior of immature, mature and hair roots of plants to explore and exploit the problem search space simultaneously. SRS divides the search space into several subspaces. It thereupon utilizes the branching and drought operations to focus on richer areas of promising subspaces while extraneous ones are not thoroughly ignored. To achieve this, the smart reactions of the SRS model are designed to act based on analyzing the heterogeneous conditions of various sections of different search spaces. In order to evaluate the performance of the SRS, it was tested on a set of known unimodal and multimodal test functions. The results were then compared with those obtained using genetic algorithms, particle swarm optimization, differential evolution and imperialist competitive algorithms and then analyzed statistically. The results demonstrated that the SRS outperformed comparative algorithms for 92% and 82% of the investigated unimodal and multimodal test functions, respectively. Therefore, the SRS is a promising nature-inspired optimization algorithm.

Application of 1H-NMR combined with qRT-PCR technology in the exploration of rosmarinic acid biosynthesis in hair roots of Salvia miltiorrhiza Bunge and Salvia castanea f. tomentosa Stib

Main conclusion Methyl jasmonate promotes the synthesis of rosmarinic acid in Salvia miltiorrhiza Bunge and Salvia castanea f. tomentosa Stib, and it promotes the latter more strongly. Abstract Salvia miltiorrhiza Bunge (SMB) is a traditional Chinese medicinal material, its water-soluble phenolic acid component rosmarinic acid has very important medicinal value. Salvia castanea f. tomentosa Stib (SCT) mainly distributed in Nyingchi, Tibet. Its pharmacological effects are similar to SMB, but its rosmarinic acid is significantly higher than the former. Methyl jasmonate (MJ) as an inducer can induce the synthesis of phenolic acids in SMB and SCT. However, the role of MJ on rosmarinic acid in SMB is controversial. Therefore, this study used SMB and SCT hair root as an experimental material and MJ as a variable. On one hand, exploring the controversial reports in SMB; on the other hand, comparing the differences in the mechanism of action of MJ on the phenolic acids in SMB and SCT. The content of related metabolites and the expression of key genes in the synthesis pathway of rosmarinic acid was analyzed by 1H-NMR combined with qRT-PCR technology. Our research has reached the following conclusions: first of all, MJ promotes the accumulation of rosmarinic acid and related phenolic acids in the metabolic pathways of SMB and SCT. After MJ treatment, the content of related components and gene expression are increased. Second, compared to SMB, SCT has a stronger response to MJ. It is speculated that the different responses of secondary metabolism-related genes to MJ may lead to different metabolic responses of salvianolic acid between the two.

The Distribution of Quetiapine and 7-Hydroxyquetiapine in Guinea Pig Hair Roots and Shafts after Repeated Administration: Exploration of the Mechanism of Drug Entry and Retention in Hair

This study investigated the distribution of quetiapine and 7-hydroxyquetiapine in guinea pig hair roots and shafts after five repeated intragastric administrations at three doses (5, 10 and 25 mg/kg) by segmental analysis to explore the mechanism of drug entry and retention in hair. Hair root samples were collected after 7, 10, 14, 21, 28 and 35 d in area A after the first dose, and a hair shaft was plucked 35 d after the first dose. The maximum concentrations of quetiapine in hair roots in the low-, medium- and high-dose groups occurred at 50, 74 and 98 h after the first administration, and the maximum concentrations were 0.71 ng/mg (range: 0.54–0.84 ng/mg), 6.72 ng/mg (range: 4.59–9.75 ng/mg) and 12.72 ng/mg (range: 10.74–15.76 ng/mg), respectively. The maximum concentrations of 7-hydroxyquetiapine in the low-, medium- and high-dose groups were 0.67 ng/mg (0.23–1.15 ng/mg), 1.07 ng/mg (0.44–1.19 ng/mg) and 3.92 ng/mg (0.656.14 ng/mg), respectively, at 26 h. The maximum concentrations of quetiapine and 7-hydroxyquetiapine in hair roots were significantly positively correlated with the dose (n = 18; r2 = 0.84; P < 0.0001 for quetiapine and n = 18; r2 = 0.61; P = 0.0001 for 7-hydroxyquetiapine). The concentrations of quetiapine and 7-hydroxyquetiapine in hair roots were higher than those in hair shafts 10 d after administration, indicating drug and metabolite entry into the hair through the roots in the first few days after administration. The highest concentrations of quetiapine in the hair shaft in the low-, medium- and high-dose groups were found at the hair ends, and 7-hydroxyquetiapine in the hair shaft showed no obvious peak concentration. Combined with previous studies, we think, by analyzing the drug concentrations in the hair roots and shaft, that the most important way for drugs to enter into and be retained in hair is that the drug enters the hair through the blood circulation from hair root, then spreads and redistributes as the hair grows.

The Study of the Yield of DNA Extracted From Hairs of Postmortem Cases

Background: Problems frequently occur from hair extraction are low concentration and impurity of DNA. Methods: Hairs from 30 postmortem cases were collected. In each case, 5 and 10 hair roots were extracted DNA by Phenol/Chloroform/Isoamyl alcohol. The amount and purity of DNA (A260/280) were detected by NanoDrop spectrophotometer. The quality of DNA was examined using polymerase chain reaction (PCR). Amount of DNA was analyzed compared with gender, age, and manner of death. Results: The amount and purity of DNA extracted from 10 hair roots were slightly better than 5 hair roots (P > .05). The mean (range) of DNA concentration was 200.4 (35.2 - 799.6) ng/µL from 10 hair roots vs 148.2 (21.7 - 571.5) ng/µL from 5 hair roots in 30 µL volume whereas the mean (range) of A260/280 was 1.61 (1.31 - 1.99) in 10 hair roots vs 1.54 (1.32 - 1.90) in 5 hair roots. The results from NanoDrop showed no difference among various gender, age, and manners of death. Conclusions: DNA can be extracted from hair by Phenol/Chloroform/Isoamyl alcohol. The quantity and quality of DNA were good enough for personal identification by PCR. DNA extracted from 10 hair roots is better than 5 hair roots.