Establishment and Optimization of Two-dimensional Electrophoresis Technique in Hydatid Fluid Proteome of Echinococcus granulosus

Abstract -The aim of this study was to establish and optimize the two-dimensional electrophoresis (2-DE) technology for hydatid fluid proteome of the Echinococcus granulosus, and obtain 2-DE map of hydatid fluid proteome. Total proteins of hydatid fluid were extracted by lyophilization. A series of important factors, such as sample preparation, protein quantities, pH range of immobilized pH gradient (IPG) strip and extraction methods, were optimized to improve the resolution and repeatability. Two-dimensional electrophoresis maps were analyzed after staining. The 2-DE profiles with high resolution and good repeatability were obtained, when the hydatid fluid dealt with ReadyPrep TM 2-D Cleanup Kit were analyzed with the established 2-DE using 400µg of quantitative loading and IPG strips pH7-10. We identified 30 protein spots using PDQuest 8.0 2D analysis software. The molecular weight of most of these proteins ranged from 43 to 97kDa and the isoelectric points of these proteins ranged from 5 to 9. An optimized 2-DE system is set up successfully in this study, electrophoresis pattern of which shows a high resolution and good repeatability, and can be used for the study of E. granulosus proteomics effectively

A heterozygous mutation in NOTCH3 in a Chinese family with CADASIL

Introduction: Cerebral autosomal-dominant arteriopathy with subcortical infarcts and leukoencephalopathy (CADASIL) is an autosomal-dominant systemic vascular disease that primarily involves small arteries. Patients with CADASIL experience migraines, recurrent ischemic strokes, cognitive decline, and dementia. The NOTCH3 gene, which is located on chromosome 19p13.12, is one of the disease-causing genes in CADASIL. Herein, we investigate the genetic and phenotypic features in a Chinese CADASIL family with heterozygous NOTCH3 mutation.Methods and Results: In the family, the proband suffered from dizziness, stroke, and cognitive deficits. Brain magnetic resonance imaging (MRI) demonstrated symmetrical white matter lesions in the temporal lobe, outer capsule, lateral ventricle, and deep brain. Whole-exome sequencing identified a known missense mutation in the proband, c.397C>T (p.Arg133Cys), which was identified in his son and granddaughter using Sanger sequencing. The proband’s younger brother and younger sister also have a history of cognitive impairment or cerebral infarction, but do not have this genetic mutation, which may highlight the impact of lifestyle on this neurological disease.Conclusion: We identified a known CADASIL-causing mutation NOTCH3 (c.397C>T, p.Arg133Cys) in a Chinese family. The clinical manifestations of mutation carriers in this family are highly heterogeneous, which is likely a common feature for the etiology of different mutations in CADASIL. Molecular genetic analyses are critical for accurate diagnosis, as well as the provision of genetic counselling for CADASIL

An Advanced Dynamic Framed-Slotted ALOHA Algorithm Based on Bayesian Estimation and Probability Response

This paper proposes an advanced dynamic framed-slotted ALOHA algorithm based on Bayesian estimation and probability response (BE-PDFSA) to improve the performance of radio frequency identification (RFID) system. The Bayesian estimation is introduced to improve the accuracy of the estimation algorithm for lacking a large number of observations in one query. The probability response is used to adjust responsive probability of the unrecognized tags to make the responsive tag number equal to the frame length. In this way, we can solve the problem of high collision rate with the increase of tag number and improve the throughput of the whole system. From the simulation results, we can see that the algorithm we proposed can greatly improve the stability of RFID system compared with DFSA and other commonly used algorithms

Polymorphisms in the Osteopontin Are Associated with Susceptibility to Ankylosing Spondylitis in a Han Chinese Population

The aim of this study was to investigate whether osteopontin (OPN) variants are associated with susceptibility to ankylosing spondylitis (AS) in a Chinese population. Polymorphisms at the 9175th position in exon 7 of OPN and rs17524488 were genotyped using direct sequencing in 186 unrelated AS patients and 188 ethnically matched healthy controls. Serum concentration of OPN was measured by enzyme-linked immunosorbent assay (ELISA) in all participants. AS patients displayed significantly higher OPN serum levels than the controls (PA in exon 7 of the OPN gene was found in this study. In healthy controls, subjects carrying the rs17524488 G/G genotype of the OPN display significantly higher OPN serum levels than the GG/GG genotype (P<0.05). Plasma OPN level is implicated as an early diagnostic marker of AS. The novel 9175th- (exon 7) position polymorphism of OPN and rs17524488 were related to susceptibility to AS in a Chinese population, the rs17524488 G/G genotype may be involved in the pathogenesis of AS, and the precise molecular mechanism underlying the influence of OPN polymorphisms on the development of AS remains to be determined in the further prospective studies

Analysis of the chemical components of hydatid fluid from Echinococcus granulosus

Introduction The aim of this study was to explore the environment of Echinococcus granulosus (E. granulosus) protoscolices and their relationship with their host. Methods Proteins from the hydatid-cyst fluid (HCF) from E. granulosus were identified by proteomics. An inductively coupled plasma atomic emission spectrometer (ICP-AES) was used to determine the elements, an automatic biochemical analyzer was used to detect the types and levels of biochemical indices, and an automatic amino acid analyzer was used to detect the types and levels of amino acids in the E. granulosus HCF. Results I) Approximately 30 protein spots and 21 peptide mass fingerprints (PMF) were acquired in the two-dimensional gel electrophoresis (2-DE) pattern of hydatid fluid; II) We detected 10 chemical elements in the cyst fluid, including sodium, potassium, calcium, magnesium, copper, and zinc; III) We measured 19 biochemical metabolites in the cyst fluid, and the amount of most of these metabolites was lower than that in normal human serum; IV) We detected 17 free amino acids and measured some of these, including alanine, glycine, and valine. Conclusions We identified and measured many chemical components of the cyst fluid, providing a theoretical basis for developing new drugs to prevent and treat hydatid disease by inhibiting or blocking nutrition, metabolism, and other functions of the pathogen

Integrating Spatial Valuation of Ecosystem Services into Protected Area Management: A Case Study of the Cangshan Nature Reserve in Dali, China

Given the importance of protected areas (PAs) in promoting the balance between conservation and sustainable development, it is crucial for planners and decision-makers to focus attention on the core areas that are of priority to PAs. However, few studies have addressed the balance between ecosystem service provision in PAs and regional development demands based on spatial valuation data. Herein, we present an integrated approach using the Cangshan Nature Reserve as a case study with the aim of identifying the core conservation areas of the reserve&mdash;the only national nature reserve in the 18-creek watershed of Cangshan&mdash;under different urbanization scenarios. The results show that the overall farmland in the watershed decreased and the architectural area increased, but the land use in the nature reserve remained stable from 1995 to 2035. With the increase in demand for water in the watershed, at least 24.3% of the Cangshan Nature Reserve should be designated as core conservation area to ensure the maintenance of sufficient water quantity and quality. This study can be used as a reference for the sustainable management of PAs based on our example of balancing ecosystem service provision and demand in a single watershed

Association of a novel frameshift variant and a known deleterious variant in MMR genes with Lynch syndrome in Chinese families

Abstract Background Lynch syndrome (LS) is the most common hereditary colorectal cancer (CRC) syndrome. This condition is characterized by germline variants in DNA mismatch repair (MMR) genes, including MLH1, MSH2, MSH6, and PMS2. In this study, we analyzed the molecular defects and clinical manifestations of two families affected with CRC and proposed appropriate individual preventive strategies for all carriers of the variant. Methods We recruited two families diagnosed with CRC and combined their family history and immunohistochemical results to analyze the variants of probands and those of other family members by using whole exome sequencing. Subsequently, gene variants in each family were screened by comparing them with the variants available in the public database. Sanger sequencing was performed to verify the variant sites. An online platform ( https://www.uniprot.org ) was used to analyze the functional domains of mutant proteins. Results A novel frameshift variant (NM_001281492, c.1129_1130del, p.R377fs) in MSH6 and a known deleterious variant (NM_000249.4:c.1731G > A, p.S577S) in MLH1 were identified in the two families with CRC. Using bioinformatics tools, we noted that the frameshift variant reduced the number of amino acids in the MSH6 protein from 1230 to 383, thereby leading to no MSH6 protein expression. The silent variant caused splicing defects and was strongly associated with LS. 5-Fluorouracil-based adjuvant chemotherapy is not recommended for patients with LS. Conclusions The novel frameshift variant (MSH6, c.1129_1130del, p.R377fs) is likely pathogenic to LS, and the variant (MLH1, c.1731G > A, p.S577S) has been further confirmed to be pathogenic to LS. Our findings underscore the significance of genetic testing for LS and recommend that genetic consultation and regular follow-ups be conducted to guide individualized treatment for cancer-afflicted families, especially those with a deficiency in MMR expression