Unknown Mutation Detection via Restriction Hybridization Method Instead of Using Next Generation Sequencing Method

In biology mutation is a change in the nucleotide sequences of the DNA of an organism Mainly there are three types of mutation: point mutation, deletion and insertions. Once the mutation has been defined allele specific oligonucleotide hybridization, amplification, heteroduplex formation method referred to as a diagnostic method some advance technique like CRISPR cas9 system is using for selected mutagenesis. Using restriction method system we can detect a mutation. Let’s say you have a DNA sample with fluorescent labeled from patient and you want to make sure that gene you are interested is in healthy gene. We can design different short fragment sequences to scan through DNA or find specific gene or mutation. The sequences scan the DNA if the sequences does not find targeted gene it does not bind to it its means that no fluorescence color appears under UV-light each different short fragment sequences is label with different colors. If the different short fragments sequence does not bind to the DNA or specific gene or area this means that there will be no color appear under UV light this part or gene will be separated from the DNA by using Restriction enzyme to do a Sanger sequencing gel electrophoresis. Result of the Sanger sequencing will provide the information about sequence of unknown part or gene of the DNA this method is easier and cost economic method instead of Next generation sequencing method [1-7].

Unknown Mutation Detection via Restriction Hybridization Method Instead of Using Next Generation Sequencing Method

In biology mutation is a change in the nucleotide sequences of the DNA of an organism mainly there are three types of mutation: point mutation, deletion and insertions. Once the mutation has been defined allele specific oligonucleotide hybridization, amplification, heteroduplex formation method referred to as a diagnostic method some advance technique like CRISPR cas9 system is using for selected mutagenesis. Using restriction method system we can detect a mutation. Let’s say you have a DNA sample with fluorescent labelled from patient and you want to make sure that gene you are interested is in healthy gene. We can design different short fragment sequences to scan through DNA or find specific gene or mutation. The sequences scan the DNA if the sequences does not find targeted gene it does not bind to it its means that no fluorescence color appears under UV-light each different short fragment sequences is label with different colors. If the different short fragments sequence does not bind to the DNA or specific gene or area this means that there will be no color appear under UV light this part or gene will be separated from the DNA by using Restriction enzyme to do a Sanger sequencing gel electrophoresis. Result of the Sanger sequencing will provide the information about sequence of unknown part or gene of the DNA this method is easier and cost economic method instead of Next generation sequencing method.

Periostin Short Fragment with Exon 17 via Aberrant Alternative Splicing Is Required for Breast Cancer Growth and Metastasis

Background: Periostin (POSTN) is a 93 kDa matrix protein that helps to regulate collagen gene expression in the extracellular matrix. POSTN overexpression is a prognostic factor in malignant cancers; however, some researchers have observed it in the stroma, whereas others have reported it on tumors. Objective: This study aimed to investigate the function of POSTN on tumors. Methods and Results: We found that POSTN in cancer cells can be detected by using an antibody against the POSTN C-terminal region exon 17 (Ex17 antibody), but not with an antibody against the POSTN N-terminal region exon 12 (Ex12 antibody) in patients with breast cancer. In a fraction secreted from fibroblasts, LC–MS/MS analysis revealed a short fragment of POSTN of approximately 40 kDa with exon 17. In addition, molecular interaction analysis showed that POSTN with exon 17, but not POSTN without exon 17, bound specifically to wnt3a, and the Ex17 antibody inhibited the binding. Conclusion: A short fragment of POSTN with exon 17, which originates in the fibroblasts, is transported to cancer cells, whereas POSTN fragments without exon 17 are retained in the stroma. The Ex17 antibody inhibits the binding between POSTN exon 17 and wnt3a.

Rapid Identification for Serotyping of African Swine Fever Virus Based on the Short Fragment of the EP402R Gene Encoding for CD2-Like Protein

The first confirmed case of African swine fever (ASF) in Vietnam was officially reported in February 2019. To date, the ASF virus (ASFV) has been detected in all 63 cities/provinces in Vietnam. In order to get a better understanding of the potential role of the EP402R gene in a grouping of ASFV serotypes, thirty ASFV sequences of EP402R genes (accession numbers: MN711757-86) from North Central Coast of Vietnam and 68 well-known references of serotype groups from previous studies were further analyzed. Interestingly, we found that a short fragment of 90 nucleotides was very typical for 8 serological groups of ASFVs. A primer set was designed to amplicon the short fragment of 90 nucleotides using the Primer3 program to establish a simplified method for the serotyping of ASFV. Our results indicated that phylogenetic analysis of the short fragment (90 nucleotides) of the EP402R gene is a very specific and useful method for ASFV serotyping when compared to the previous method using a long fragment (816 nucleotides) of this gene and well-known serotype references based on haemadsorption inhibition (HAI) assay.

Using environmental DNA methods to survey for rare groundwater fauna: Detection of an endangered endemic cave crayfish in northern Alabama

The conservation and management of subterranean biodiversity is hindered by a lack of knowledge on the true distributions for many species, e.g., the Wallacean shortfall. In recent years, several studies have demonstrated the potential of environmental DNA (eDNA) as an effective approach to detect and monitor biodiversity, including rare, threatened, and endangered taxa. However, there are few eDNA studies of groundwater fauna. Here we report the results of the development and implementation of an eDNA assay targeting a short fragment of the mitochondrial CO1 locus of a critically imperiled cave crayfish, the Sweet Home Alabama Cave Crayfish (Cambarus speleocoopi), known from just four cave systems in the Interior Plateau karst region of northern Alabama. We detected C. speleocoopi DNA from water samples collected at 5 of 16 sites sampled (caves and springs), including two historical sites as well as three additional and potentially new sites in Marshall County, Alabama. All three of these sites were within 2 km of historical sites. Our study is the first to detect a groundwater crustacean in the Interior Plateau karst region. Additionally, our study contributes to the growing literature that eDNA is a viable complementary tool for detection and monitoring of a fauna that is difficult to survey and study using traditional approaches.

Biomechanical Comparison of a Notched Head Locking T-Plate and a Straight Locking Compression Plate in a Juxta-Articular Fracture Model

Objective This investigation compared the biomechanical properties of a 2.0 mm locking compression notched head T-plate (NHTP) and 2.0 mm straight locking compression plate (LCP), in a simple transverse juxta-articular fracture model. Study Design Two different screw configurations were compared for the NHTP and LCP, modelling short (configuration 1) and long working length (configuration 2). Constructs were tested in compression, perpendicular and tension non-destructive four point bending and torsion. Plate surface strain was measured at 12 regions of interest (ROI) using three-dimensional digital image correlation. Stiffness and strain were compared between screw configurations within and between each plate. Results The LCP was stiffer than the NHTP in all three planes of bending and torsion (p < 0.05). The NHTP had greater strain than the LCP during compression bending and torsion at all ROI (p < 0.0005). The short working length was stiffer in all three planes of bending and in torsion (p < 0.05) than the longer working length for both plates. The long working length showed greater strain than the short working length at most ROI. Conclusion In this experimental model, a 2.0 mm LCP with two screws in the short fragment was significantly stiffer and had lower plate strain than a 2.0 mm NHTP with three screws in the short fragment. Extending the working length significantly reduced construct stiffness and increased plate strain. These findings may guide construct selection.

Effect of Plate Type and Working Length on a Synthetic Compressed Juxta-Articular Fracture Model

Objective This investigation compared the biomechanical properties of a 2.0 mm locking compression notched head T-plate (NHTP) and 2.0 mm straight locking compression plate (LCP), in a compressed, short, juxta-articular fragment fracture model. Methods Two different screw configurations were compared for the NHTP and LCP, modelling short (configuration 1) and long working length (configuration 2). Constructs were tested in compression, perpendicular and tension four-point bending and torsion. Plate surface strain was measured at 12 regions of interest using three-dimensional digital image correlation. Stiffness and strain were compared. Results The LCP was stiffer than the NHTP in all three planes of bending (p < 0.05). The NHTP was stiffer than the LCP in torsion (p < 0.05). The NHTP had greater strain than the LCP during compression bending and torsion (p < 0.0005). The short working length NHTP was stiffer in all three planes of bending and in torsion (p < 0.05) than the longer working length. The short working length LCP was stiffer in compression bending and in torsion (p < 0.05) than the longer working length. The long working length showed greater strain than the short working length at multiple regions of interest. Conclusion In this experimental model of a compressed transverse fracture with a juxta-articular 9 mm fragment, a 2.0 mm LCP with two hybrid screws in the short fragment was stiffer than a 2.0 mm NHTP with three locking screws in the short fragment in three planes of bending but not torsion. Extending the working length of each construct reduced construct stiffness and increased plate strain.

Rejestr dworzan konnych Zygmunta I Starego wysłanych do walki z Krzyżakami pustoszącymi Mazowsze na przełomie lipca i sierpnia 1520 roku

The mounted courtiers of Sigismund I the Old, in addition to their representative function, were a small, trained reserve formation remaining permanently at the side of the king, who could issue them an order at any time. During the so-called Prussian war of 1519–1521, the size of this group reached its peak in the summer of 1520 in Toruń (539 horses in 121 court detachments – pocztach dworskich). During the fights with the Teutonic Knights, the courtiers were constantly responsible for the safety of the monarch in his place of residence, but smaller groups took part in military actions. The analysis of a short fragment of source material included in the manuscript no. 49 (Main Archive of Old Files, Crown Treasury Archive, Section 1: Royal Accounts) allowed to make a detailed description of a small banner, consisting of 69 horses in 15 retinues, taking part in the fight against the Teutonic Knights’ plundering raids to northern Mazovia at the turn of July and August 1520. The composition of the unit and the nature and size of the retinues were discussed, as well as the issue of commanding the group, the costs incurred for its action and, based on Marian Biskup’s research, the battle route was presented. The presented example can be taken as a starting point for a discussion on the military organization of the court. It can be assumed that the king or the marshal, as the court’s superior, would decide which of the individual retinues of mounted courtiers would form one or more court banners that functioned simultaneously and independently. Their number and composition were probably determined by the tasks envisaged for the units.