Traditional Morphological Methods Research Articles

Assessing arthropod biodiversity with DNA barcoding in Jinnah Garden, Lahore, Pakistan.

Previous difficulties in arthropod taxonomy (such as limitations in conventional morphological approaches, the possibility of cryptic species and a shortage of knowledgeable taxonomists) has been overcome by the powerful tool of DNA barcoding. This study presents a thorough analysis of DNA barcoding in regards to Pakistani arthropods, which were collected from Lahore's Jinnah Garden. The 88 % (9,451) of the 10,792 specimens that were examined were able to generate DNA barcodes and 83% (8,974) of specimens were assigned 1,361 barcode index numbers (BINs). However, the success rate differed significantly between the orders of arthropods, from 77% for Thysanoptera to an astounding 93% for Diptera. Through morphological exams, DNA barcoding, and cross-referencing with the Barcode of Life Data system (BOLD), the Barcode Index Numbers (BINs) were assigned with a high degree of accuracy, both at the order (100%) and family (98%) levels. Though, identifications at the genus (37%) and species (15%) levels showed room for improvement. This underscores the ongoing need for enhancing and expanding the DNA barcode reference library. This study identified 324 genera and 191 species, underscoring the advantages of DNA barcoding over traditional morphological identification methods. Among the 17 arthropod orders identified, Coleoptera, Diptera, Hemiptera, Hymenoptera, and Lepidoptera from the class Insecta dominated, collectively constituting 94% of BINs. Expected malaise trap Arthropod fauna in Jinnah Garden could contain approximately 2,785 BINs according to Preston log-normal species distribution, yet the Chao-1 Index predicts 2,389.74 BINs. The Simpson Index of Diversity (1-D) is 0.989, signaling high species diversity, while the Shannon Index is 5.77, indicating significant species richness and evenness. These results demonstrated that in Pakistani arthropods, DNA barcoding and BOLD are an invaluable tool for improving taxonomic understanding and biodiversity assessment, opening the door for further eDNA and metabarcoding research.

Molecular based identification and phylogenetic relationship by using cytochrome b gene of Pangasius pangasius.

Molecular appraoch for identification of unknown species by using Cytochrome b gene is an effective and reliable as compared with morphological based identification. For DNA barcoding universal molecular genes were used to identify the species. Cytochrome b is a specific gene used for identification purpose. DNA barcoding is a reliable and effective method compared to the different traditional morphological methods of specie identification. So,in the present study which was conducted to identify the species, a total of 50 fish samples were collected from five different sites. DNA was extracted by using the Phenol Chloroform method from muscle tissue. Five sequences were sequenced (one from each site), analyzed, and identified specific species as Pangasius pangasius. Identified sequences were variable in length from 369 bp (Site 1), 364 bp (Site 2), 364 bp (Site 3), 352 bp (Site 4), and 334 bp (Site 5). Identity matches on the NCBI database confirmed the specific specie as P. pangasius. A distancing tree was drawn to show maximum likelihood among the same and different species. Yet, in many cases fishes on diverse development stages are difficult to identify by morphological characters. DNA-based identification methods offer an analytically powerful addition or even an alternative tool for species identification and phylogenetic study. This work intends to provide an updated and extensive overview on the DNA based methods for fish species identification by using Cytochrome b gene as targeted markers for identification purpose.

Biochemical indicators used in ecological monitoring of surface waters

The work is devoted to the search for promising test systems for ecological monitoring of surface waters by comparing the effectiveness of detecting low concentrations of various types of pesticides in the aquatic environment by traditional morphological and biochemical research methods.
 Allium cepa L. and the hydatophyte plant Elodea canadensis Michx. were used as test objects because it is immersed in the aquatic environment during its entire life cycle and is in the most complete contact with all the toxicants present in the water.
 It was found that the traditionally used test reactions of plants cannot always reflect a potentially dangerous exceedance of the MPC of toxicants in the aquatic environment.
 The possibility of using catalase activity as a sensitive bioassay for the presence of low concentrations of herbicides with different mechanisms of action in the water environment was investigated.
 Correlation analysis has been conducted, and Pearson's coefficients have been calculated to quantify the relationship between the concentration of herbicides present in water and the degree of manifestation of test reactions by the research objects.
 The highest level of correlation between the increase in the concentration of herbicides in water and the test reactions of elodea was demonstrated by biochemical indicators, and the higher significance of the correlation was observed at 4-day exposure of plants in the presence of the herbicide.

Blood supply to the stomach of Yorkshire pigs at the stages of postnatal ontogenesis

The development of the agro–industrial complex of the Russian Federation is one of the priorities in agriculture. Pig breeding is one of the most profitable branches of animal husbandry. This is due to the fact that pigs have a number of biological features that are successfully used to produce products (pork, bacon, bacon). With the intensification of pig breeding, special attention is paid to scientific approaches to improving pig feeding. In this regard, we set ourselves the goal of the study – to study the arterial vascularization of the stomach in Yorkshire piglets and to determine the morphometric data of the organ at the stages of postnatal ontogenesis. The study was conducted at the Department of Animal Anatomy of the St. Petersburg State University of Veterinary Medicine. Cadaver material for the study was delivered to the Department of Animal Anatomy of the St. Petersburg State University of Veterinary Medicine from the Idavang Agro pig breeding complex in Nurma village, Tosnensky district of the Leningrad Region. The study was conducted in three age groups, according to the periodization of the life of pigs. To achieve this task, traditional morphological research methods were used, namely: fine anatomical dissection, photographing, vasorentgenography with morphometry in the "RadiAnt" program. During the study, it was found that arterial gastric vascularization in Yorkshire piglets at the stages of postnatal ontogenesis is carried out due to the hepatic and splenic arteries, which are branches of the abdominal artery. The branches of the arteries of the first order branch in a loose type. When conducting a comparative analysis of the morphometric characteristics of the hepatic and splenic arteries and their branches, we came to the conclusion that the diameter of the hepatic artery will prevail over the splenic, and their branches of the first order are diametrically opposed to each other.

Relevance of molecular systematics in insect pest management

Relevance of molecular systematics in insect pest management

Research on the dentification of Panax ginseng and Panax quinquefolium using catalysed hairpin assembly technology.

Panax ginseng and Panax quinquefolium are traditional Chinese herb medicines and similar in morphology and some chemical components but differ in drug properties, so they cannot be mixed. However, the processed products of them are often sold in the form of slices, powder, and capsules, which are difficult to identify by traditional morphological methods. Furthermore, an accurate evaluation of P. ginseng, P. quinquefolium and the processed products have not been conducted. This study aimed to establish a catalysed hairpin assembly (CHA) identification method for authenticating products made from P. ginseng and P. quinquefolium based on single nucleotide polymorphism (SNP) differences. By analysing the differences of SNP in internal transcribed spacer 2 (ITS2) in P. ginseng and P. quinquefolium to design CHA-specific hairpins. Establish a sensitive and efficient CHA method that can identify P. ginseng and P. quinquefolium, use the sequencing technology to verify the accuracy of this method in identifying Panax products, and compare this method with high-resolution melting (HRM). The reaction conditions of CHA were as follows: the ratio of forward and reverse primers, 20:1; hairpin concentration, 5 ng/μL. Compared with capillary electrophoresis, this method had good specificity and the limit of detection was 0.5ng/μL. The result of Panax product identification with CHA method were coincidence with that of the sequencing method; the positive rate of CHA reaction was 100%. This research presents an effective identification method for authenticating P. ginseng and P. quinquefolium products, which is helpful to improve the quality of Panax products.

Capturing drifting species and molecules—Lessons learned from integrated approaches to assess marine metazoan diversity in highly dynamic waters

AbstractMarine community diversity surveys require a reliable assessment to estimate ecosystem functions and their dynamics. For these, non‐invasive environmental DNA (eDNA) metabarcoding is increasingly applied in zoological studies to complement or even replace traditional morphological identification methods. However, uncertainties remain about the accuracy of the diversity detected with eDNA to capture the actual diversity in the field. Here, we validate the reliability of eDNA metabarcoding in identifying metazoan biodiversity in highly dynamic marine waters of the North Sea. We analyzed biodiversity from water (eDNA) and zooplankton samples with cytochrome c oxidase subunit 1 (COI) and 18S rRNA (18S) metabarcoding at Helgoland Roads and validated the optimal molecular resolution by morphological and molecular zooplankton identification (metabarcoding) with the result of merely a few false‐negative detections. eDNA and zooplankton metabarcoding resolved 354 species from all major and in total 16 metazoan phyla. This molecular genetic species inventory overlapped by 95.9% (COI) and 81.9% (18S) with published inventories of local, morphologically identified species, among them neozoa and rediscovered species. Even though half of all species were detected by both eDNA and zooplankton metabarcoding, the methods differed significantly in their detected diversity. eDNA metabarcoding performed very well in cnidarians and annelids, whereas zooplankton metabarcoding identified higher numbers of fish and malacostraca. Species assemblages significantly differed between the individual sampling events and the cumulative number of identified species increased steadily over the sampling period and did not reach saturation. About a third of the species were detected only once while a core community of 22 species was identified continuously. Our study confirms eDNA metabarcoding to be a powerful tool to identify and analyze North Sea fauna in highly dynamic waters and we recommend investing in high sampling efforts by repetitive sampling and replication using at least 0.45 μm filters to increase filtration volume.

Comparative Skull and Mandible Geometric Morphometrics of Two Species of Mice, Mus domesticus and Mus macedonicus (Muridae, Rodentia) in Turkey

Using a geometric morphometrics approach, we examined shape and size variations of skull and mandible bone of two evolutionarily distantly related mice from Turkey: Mus domesticus and Mus macedonicus. PCA analyses revealed overlap in dorsal cranium and mandible shapes of both species, consistent with previous traditional morphological methods. The skull of M. macedonicus seems to be larger in size than M. domesticus according to box-plot analyses of centroid size values, however there is no obvious difference for the mandible. No difference was observed between sexes in either of the characters. We suggest that future studies focus on dental characteristics and also consider the variation among local populations and ecological variables.

Ultra-deep sequencing of 45S rDNA to discern intragenomic diversity in three Chrysodeixis species for molecular identification

Species identification is necessary to prevent introductions of exotic plant pests through global trade. Many of these pests are understudied and lack publicly available DNA sequence data on which rapid molecular identification methods can be based. One such lineage is the genus Chrysodeixis, which includes three species of potential concern for United States trade initiatives: C. includens, C. chalcites, and C. eriosoma. Here we describe a method to generate robust 45S rDNA profiles using long read sequencing in order to clarify evolutionary relationships and develop a real-time PCR identification technique. Such an identification tool will be useful in rapidly differentiating between Chrysodeixis species of quarantine concern where traditional morphological identification methods are insufficient. Molecular methods such as this greatly reduce the time spent identifying each specimen, allow for detection of eDNA, vastly increase throughput, and increase the probability of detection. The methods presented here will be generally adaptable to any understudied lepidopteran taxa that necessitates a molecular diagnostic assay and, with adjustment or testing of the primers, could be applied to any group for which development of rDNA profiles in a benchtop setting would prove useful.

Systematic review and meta-analysis: does pre-implantation genetic testing for aneuploidy at the blastocyst stage improve live birth rate?

PurposeTo establish if preimplantation genetic testing for aneuploidy (PGT-A) at the blastocyst stage improves the composite outcome of live birth rate and ongoing pregnancy rate per embryo transfer compared to conventional morphological assessment.MethodsA systematic literature search was conducted using PubMed, EMBASE and Cochrane database from 1st March 2000 until 1st March 2022. Studies comparing reproductive outcomes following in vitro fertilisation using comprehensive chromosome screening (CCS) at the blastocyst stage with traditional morphological methods were evaluated.ResultsOf the 1307 citations identified, six randomised control trials (RCTs) and ten cohort studies fulfilled the inclusion criteria. The pooled data identified a benefit between PGT-A and control groups in the composite outcome of live birth rate and ongoing pregnancy per embryo transfer in both the RCT (RR 1.09, 95% CI 1.02–1.16) and cohort studies (RR 1.50, 95% CI 1.28–1.76). Euploid embryos identified by CCS were more likely to be successfully implanted amongst the RCT (RR 1.20, 95% CI 1.10–1.31) and cohort (RR 1.69, 95% CI 1.29–2.21) studies. The rate of miscarriage per clinical pregnancy is also significantly lower when CCS is implemented (RCT: RR 0.73, 95% CI 0.56–0.96 and cohort: RR 0.48, 95% CI 0.32–0.72).ConclusionsCCS-based PGT-A at the blastocyst biopsy stage increases the composite outcome of live births and ongoing pregnancies per embryo transfer and reduces the rate of miscarriage compared to morphological assessment alone. In view of the limited number of studies included and the variation in methodology between studies, future reviews and analyses are required to confirm these findings.

Histological features of the bladder wall in yorkshire piglets in the age aspect

Yorkshire pigs occupy one of the first places in livestock farms in different parts of the world. This is due to the fact that breeding boars of this breed are widely used by farmers and owners not only of large livestock complexes, but also by private farms for crossing with other breeds of adapted pigs to increase the meat indicators of offspring. Pigs of this breed are very sensitive to sudden changes in ambient temperature, often suffer from overheating or hypothermia, which can affect the health of internal organs and body systems and can lead to diseases of the urinary system in general and in particular the bladder. The purpose of our study is to study the features of the histological structure of the bladder wall in Yorkshire piglets in the age aspect. Cadaver material for the study was delivered to the Department of Animal Anatomy of the Federal State Budgetary Educational Institution of the Russian Academy of Sciences from the Idavang Agro pig breeding complex in Nurma village, Tosnensky district of the Leningrad Region. The study was conducted in three age groups, according to the periodization of pig life: the first stage of development – newborn piglets (1-4 days of the postnatal period); the second stage of development – the first dairy (5-21 days of the postnatal period); the third stage of development – the second dairy (22-45 days of the postnatal period). From the first stage of development, newborn one-day-old piglets were used; from the second stage – animals 10-14 days; from the third – piglets 28-30 days of the postnatal period of ontogenesis. The average body weight of piglets of group 1 is 550 g.; Group 2 is 1800 g.; Group 3 is 3000 g. In total, five piglets were inherited in each age group. The age was determined by the bonus cards and from the words of the chief veterinarian of the farm. To achieve this task, a set of traditional morphological research methods was used: fine anatomical dissection, histological, photographing and morphometry. Histological examination revealed that in Yorkshire piglets, the bladder wall has a typical histological structure and consists of mucous, muscular, and serous membranes. Morphometric indicators of histological structures of the bladder wall were determined and it was concluded that these values of thickness and diameter in males slightly exceed those in females.

Applicability and Advantage of Mitochondrial Metagenomics and Metabarcoding in Spider Biodiversity Survey

Spiders are an extraordinary animal group with extremely high diversity in species, morphology, and behavior. Accurate estimation of species diversity and community composition is essential in spider ecological studies as well as applications of biodiversity surveys and monitoring. However, spider biodiversity surveys still largely utilize the morphology-based approach, which is often time-consuming and highly dependent on taxonomic experts. In this study, we tested the applicability of mitochondrial metagenomic and metabarcoding methods in the biodiversity survey of spiders. Local mitogenome and barcode databases of 62 reference species were built using next-generation sequencing and Sanger sequencing technologies. The performances of mitochondrial metagenomics, metabarcoding, and morphological methods were compared for five plot samples of spiders. The results show that the molecular methods (mitochondrial metagenomics and metabarcoding) have a higher species detection rate compared with the traditional morphological biodiversity method, which is largely due to their capability of incorporating the large proportion of immature specimens in each plot into the biodiversity assessment. Our study demonstrates the great potential of molecular approaches in advancing spider biodiversity and community ecology studies and suggests that by combining both mitochondrial metagenomic and metabarcoding methods we could provide more accurate and reliable biodiversity assessment for spiders.

The phylogenetic structure and coalescent species delimitation of an endemic trapdoor spider genus, Stasimopus (Araneae, Mygalomorphae, Stasimopidae) in the Karoo region of South Africa

The Karoo region of South Africa is a unique and sensitive ecosystem which is facing pressure for development due to economic incentives such as mining, farming and shale gas exploration. The species diversity of many taxa in the area is largely unknown. A phylogenetic analysis of the cork-lid trapdoor spider genus, Stasimopus (Stasimopidae) was undertaken in order to gain insight into the relationships between the species that may be present in the area. The species within Stasimopus are challenging to identify and define using traditional morphological methods due to a high degree of morphological conservatism within the genus. For this reason, multiple coalescent based species delimitation methods were used to attempt to determine the species present for Stasimopus in the region which was tested against the morphological identifications and genetic clades (based on CO1, 16S and EF-1ɣ). We tested single-locus methods Automatic Barcode Gap Discovery (ABGD), Bayesian implementation of Poisson Tree Processes (bPTP) and General Mixed Yule- Coalescent (GMYC), as well as multi-locus Brownie. The phylogenetic analysis of Stasimopus in the Karoo showed that there is a high degree of genetic diversity within the genus. The species delimitation results proved unfruitful for the genus, as they appear to delimit population structure rather than species for most methods. Alternative methods should be investigated to aid in the identification of the species in order truly understand the species diversity of the genus.

Gerhardtia tomentosa and Ossicaulis borealis (Agaricales, Lyophyllaceae)-Two new species from northeast China.

Gerhardtia and Ossicaulis are two genera within the family Lyophyllaceae, which show an apparently poor species diversity worldwide. During the field investigation on wild macrofungi, six interesting collections within Gerhardtia and Ossicaulis genera are discovered in the northeastern China. To identify whether these collections of Gerhardtia and Ossicaulis are novel species, we performed phylogenetic analyzes using the following DNA regions: the internal transcribed spacer (ITS) region and the large subunit nuclear ribosomal RNA (nrLSU) region. Moreover, a traditional morphological method also be conducted based on both the macro-morphological and micro-morphological features. The results indicated that these collections tested formed two independent lineages in each genus with a high support. In addition, they can easily be separated from all other taxa of the two genera in morphology. Based on the combination of morphological and molecular data, Gerhardtia tomentosa and Ossicaulis borealis, are confirmed as two new species to science. This study provided a theoretical basis is for the two lyophylloid genera and indicated that the biodiversity resources of northeastern China might be underestimated.

Rapid Morphological Measurement Method of Aortic Dissection Stent Based on Spatial Observation Point Set.

Post-operative stent morphology of aortic dissection patients is important for performing clinical diagnosis and prognostic assessment. However, stent morphologies still need to be manually measured, which is a process prone to errors, high time consumption and difficulty in exploiting inter-data associations. Herein, we propose a method based on the stepwise combination of basic, non-divisible data sets to quickly obtain morphological parameters with high accuracy. We performed the 3D reconstruction of 109 post-operative follow-up CT image data from 26 patients using mimics software. By extracting the spatial locations of the basic morphological observation points on the stent, we defined a basic and non-reducible set of observation points. Further, we implemented a fully automatic stent segmentation and an observation point extraction algorithm. We analyzed the stability and accuracy of the algorithms on a test set containing 8 cases and 408 points. Based on this dataset, we calculated three morphological parameters of different complexity for the different spatial structural features exhibited by the stent. Finally, we compared the two measurement schemes in four aspects: data variability, data stability, statistical process complexity and algorithmic error. The statistical results of the two methods on two low-complexity morphological parameters (spatial position of stent end and vascular stent end-slip volume) show good agreement (n = 26, P1, P2 < 0.001, r1 = 0.992, r2 = 0.988). The statistics of the proposed method for the morphological parameters of medium complexity (proximal support ring feature diameter and distal support ring feature diameter) avoid the errors caused by manual extraction, and the magnitude of this correction to the traditional method does not exceed 4 mm with an average correction of 1.38 mm. Meanwhile, our proposed automatic observation point extraction method has only 2.2% error rate on the test set, and the average spatial distance from the manually marked observation points is 0.73 mm. Thus, the proposed method is able to rapidly and accurately measure the stent circumferential deflection angle, which is highly complex and cannot be measured using traditional methods. The proposed method can significantly reduce the statistical observation time and information processing cost compared to the traditional morphological observation methods. Moreover, when new morphological parameters are required, one can quickly and accurately obtain the target parameters by new "combinatorial functions." Iterative modification of the data set itself is avoided.

Real-time PCR assay for Tetranychus evansi and multiplex with three other Tetranychus species

Spider mites are significant agricultural and horticultural pests. Among them, Tetranychus urticae Koch (Acari: Tetranychidae) is the most important one and has a cosmopolitan distribution. The tomato red spider mite (TRSM), T. evansi Baker and Pritchard is a worldwide pest of solanaceous crops and has become a major invasive agricultural pest. Tetranychus evansi originates from South America and has recently invaded many other parts of the world, including countries of Africa, Europe, Asia and North America. Recently, it has spread to Australia and New Zealand. TRSM has a high intrinsic rate of increase and completes a generation in less than 15 days. It is reported that T. evansi has replaced T. uritcae and other endemic spider mites as the dominant pest and also colonised new host plants in the invaded countries. Furthermore, T. urticae, T. ludeni and T. lambi shared the same host as T. evansi. Tetranychus species are difficult to distinguish using traditional morphological methods, and the molecular techniques such as mitochondrial COI and ITS sequences have greatly assisted in the diagnosis of those species. However, the methods include post-PCR steps and are time-consuming. In contrast, real-time PCR assays which omit those post-PCR steps can provide rapid detection. The real-time PCR assay to identify T. urticae, T. ludeni and T. lambi were already developed and validated by us (Li et al., 2015) and by Australian scientists (Chen et al., 2020), but no assay targeting T. evansi is available. Therefore, we developed and validated a TaqMan real-time PCR protocol for rapid detection of T. evansi, by targeting ITS1 gene region. The assay specificity was tested against the closely related species, i.e., T. collyerae, T. lambi, T. ludeni, T. kanzawai, T. parakanzawai, pueraicola, T. truncatus, T. turkestani and T. urticae. Plasmid DNA containing ITS1 sequence of T. evansi was used as standards for sensitivity tests of the real-time PCR assay. The detection limit of the assay is 10 copy of the target region, indicating that it can detect DNA from different life stage of the target species, i.e. an egg, a larva, a nymph or an adult. Furthermore, multiplex assay targeting T. evansi, T. ludeni, T. lambi and T. urticae have been validated for detection of the four species. The validation showed that the assay against T. evansi can run as singleplex and also as multiplex by targeting the four species simultaneously. This multiplex assay has been used in the surveillance programme in New Zealand and demonstrated its efficiency in detection of the target species. Moreover, the assay will be very useful for the quick identification of intercepted specimens at border and post border.

An Unsupervised Machine Learning Method for Electron–Proton Discrimination of the DAMPE Experiment

Galactic cosmic rays are mostly made up of energetic nuclei, with less than 1% of electrons (and positrons). Precise measurement of the electron and positron component requires a very efficient method to reject the nuclei background, mainly protons. In this work, we develop an unsupervised machine learning method to identify electrons and positrons from cosmic ray protons for the Dark Matter Particle Explorer (DAMPE) experiment. Compared with the supervised learning method used in the DAMPE experiment, this unsupervised method relies solely on real data except for the background estimation process. As a result, it could effectively reduce the uncertainties from simulations. For three energy ranges of electrons and positrons, 80–128 GeV, 350–700 GeV, and 2–5 TeV, the residual background fractions in the electron sample are found to be about (0.45 ± 0.02)%, (0.52 ± 0.04)%, and (10.55 ± 1.80)%, and the background rejection power is about (6.21 ± 0.03) × 104, (9.03 ± 0.05) × 104, and (3.06 ± 0.32) × 104, respectively. This method gives a higher background rejection power in all energy ranges than the traditional morphological parameterization method and reaches comparable background rejection performance compared with supervised machine learning methods.

Candidate biomarkers in brown adipose tissue for post-mortem diagnosis of fatal hypothermia.

Post-mortem diagnosis of fatal hypothermia (FHT) is challenging in forensic practice because traditional morphological and biochemical methods lack specificity. Recent studies have reported that brown adipose tissue (BAT) is activated during cold-induced non-shivering thermogenesis in mammals, but BAT has not been used to diagnose FHT. The aim of this study was to identify novel biomarkers in BAT for FHT based on morphological changes and differential protein expression. Two FHT animal models were created by exposing mice to 4 or -20 °C at 50% humidity. Morphologically, the unilocular lipid droplet content was significantly increased in BAT of FHT model mice compared with that of control mice. Proteomics analysis revealed a total of 283 and 266 differentially expressed proteins (DEPs) between the 4 or -20 °C FHT subgroups and control group, respectively. In addition, 140 proteins were shared between the FHT subgroups. GO and KEGG analyses revealed that the shared DEPs were mainly enriched in pathways associated with metabolism, oxidative phosphorylation, and thermogenesis. Further screening (|log2FC| > 1.6, q-value (FDR) < 0.05) identified GMFB, KDM1A, DDX6, RAB1B, SHMT-1, CLPTM1, and LMF1 as candidate biomarkers of FHT. Subsequent validation experiments were performed in FHT model mice using classic immunohistochemistry and western blotting. RAB1B and GMFB expression was further verified in BAT specimens from human cases of FHT. The results demonstrate that BAT can be used as a target organ for FHT diagnosis employing RAB1B and GMFB as biological markers, thus providing a new strategy for the post-mortem diagnosis of FHT in forensic practice.

Targeted Metabolomics Provide Chemotaxonomic Insights of Medicago ruthenica, with Coupled Transcriptomics Elucidating the Mechanism Underlying Floral Coloration.

Medicago ruthenica, a wild legume forage widely distributed in the Eurasian steppe, demonstrates high genetic and phenotypic variation. M. ruthenica with a purely yellow flower (YFM), differing from the general phenotype of M. ruthenica with a purple flower (PFM), was recently discovered. The similar characteristics of YFM with Medicago falcata have led to conflicting opinions on its taxonomy using traditional morphological methods. The lack of chemotaxonomy information about M. ruthenica species and the unclear flower coloration mechanisms have hampered their study. Here, we investigated M. ruthenica using targeted metabolomics based on the chemotaxonomy method and elaborated the floral coloration mechanisms using transcriptomics. The identified flavonoids were the same types, but there were different contents in YFM and PFM, especially the contents of cyanidin-3-O-glucoside (C3G), an anthocyanin that causes the purple-reddish color of flowers. The over-accumulation of C3G in PFM was 1,770 times more than YFM. Nineteen anthocyanin-related genes were downregulated in YFM compared with their expression in PFM. Thus, YFM could be defined as a variety of M. ruthenica rather than a different species. The loss of purple flower coloration in YFM was attributed to the downregulation of these genes, resulting in reduced C3G accumulation. The taxonomic characteristics and molecular and physiological characteristics of this species will contribute to further research on other species with similar external morphologies.

Bearing Fault Feature Extraction Method Based on Enhanced Differential Product Weighted Morphological Filtering.

Aimed at the problem of fault characteristic information bearing vibration signals being easily submerged in some background noise and harmonic interference, a new algorithm named enhanced differential product weighted morphological filtering (EDPWMF) is proposed for bearing fault feature extraction. In this method, an enhanced differential product weighted morphological operator (EDPWO) is first constructed by means of infusing the differential product operation and weighted operation into four basic combination morphological operators. Subsequently, aiming at the disadvantage of the parameter selection of the structuring element (SE) of EDPWO depending on artificial experience, an index named fault feature ratio (FFR) is employed to automatically determine the flat SE length of EDPWO and search for the optimal weighting correlation factors. The fault diagnosis results of simulation signals and experimental bearing fault signals show that the proposed method can effectively extract bearing fault feature information from raw bearing vibration signals containing noise interference. Moreover, the filtering result obtained by the proposed method is better than that of traditional morphological filtering methods (e.g., AVG, STH and EMDF) through comparative analysis. This study provides a reference value for the construction of advanced morphological analysis methods.