GALNT9 enrichment attenuates MPP-induced cytotoxicity by ameliorating protein aggregations containing α-synuclein and mitochondrial dysfunction.

Background: GALNTs (UDP-GalNAc; polypeptide N-acetylgalactosaminyltransferases) initiate mucin-type O-GalNAc glycosylation by adding N-GalNAc to protein serine/threonine residues. Abnormalities in O-GalNAc glycosylation are involved in various disorders such as Parkinson's disease (PD), a neurodegenerative disorder. GALNT9 is potentially downregulated in PD patients.

Quantitatively Evaluating Interactions between Patient-Derived Organoids and Autologous Immune Cells by Microfluidic Chip.

The coculture of patient-derived tumor organoids (PDOs) and autologous immune cells has been considered as a useful surrogate of tumor-immune environment. However, the immune interactions between PDOs and autologous immune cells, including immune-mediated killing behaviors and immune-related cytokine variations, have yet to be quantitatively evaluated. This study presents a microfluidic chip for quantifying interactions between PDOs and autologous immune cells (IOI-Chip).

Deficiency of polypeptide N-acetylgalactosamine transferase 9 contributes to a risk for Parkinson's disease via mitochondrial dysfunctions.

Polypeptide N-acetylgalactosamine transferase 9 (GALNT9) catalyzes the initial step of mucin-type O-glycosylation via linking N-acetylgalactosamine (GalNAc) to serine/threonine in a protein. To unravel the association of GALNT9 with Parkinson's disease (PD), a progressive neurodegenerative disorder, GALNT9 levels were evaluated in the patients with PD and mice treated with 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine, and statistically analyzed based on the GEO datasets of GSE114918 and GSE216281. Glycoproteins with exposing GalNAc were purified using lectin affinity chromatography and identified by LC-MS/MS.

Exploratory application of an integrated topic-based curriculum in biochemistry experimental teaching.

Biochemistry, a complicated basic course in health sciences, plays a fundamental role in describing pathogenic mechanism of illness in molecular terms, and is required to be learned for all medical students. Due to various obstacles to biochemistry learning and teaching, there is a pressing issue of curriculum reform to arouse the student's enthusiasm in biochemistry learning. In this study, an integrated topic-based biochemistry training program (TBBTP) combining the traditional lectures, question-based learning and experimental practices, was introduced in biochemistry teaching.

Identification of ADP/ATP Translocase 1 as a Novel Glycoprotein and Its Association with Parkinson's Disease.

Protein glycosylation plays a crucial role in central nervous system, and abnormal glycosylation has major implications for human diseases. This study aims to evaluate an etiological implication of the variation in glycosylation for Parkinson's disease (PD), a neurodegenerative disorder. Based on a PD mouse model constructed by the intraperitoneal injection with 1-Methyl-4-phenyl-1,2,3,6-tetrahydropyridine, glycosylation variation was accessed using biotinylated lectin of dolichos biflorus agglutinin (DBA) specific for the exposed N-acetylgalactosamine linked to glycoprotein.

ADP/ATP translocase 1 protects against an α-synuclein-associated neuronal cell damage in Parkinson's disease model.

Background: ADP/ATP translocase 1 (ANT1) is involved in the exchange of cytosolic ADP and mitochondrial ATP, and its defection plays an important role in mitochondrial pathogenesis. To reveal an etiological implication of ANT1 for Parkinson's disease (PD), a neurodegenerative disorder, a mouse model treated with 1-Methyl-4-phenyl-1,2,3,6-tetrahydropyridine and neuroblastoma cell model induced by 1-methyl-4-pehny1-pyridine were utilized in this study.

Results: The tissue-specific abundance in ANT1 in mouse brains was accessed using the analysis of Western blot and immunohistochemistry.

The mesenchymal property of mouse mammary anlagen repopulating cell population is associated with its stemness.

During early embryogenesis, mammary glands are derived from surface ectoderm and their morphogenesis is controlled by mammary stem cells (MaSCs) and epithelial-mesenchymal transition (EMT). Mammary anlagen stage (E13.5-15.

Proteomic and Morphologic Evidence for Taurine-5-Bromosalicylaldehyde Schiff Base as an Efficient Anti-Mycobacterial Drug.

, a causative pathogen of tuberculosis (TB), still threatens human health worldwide. To find a novel drug to eradicate this pathogen, we tested taurine-5- bromosalicylaldehyde Schiff base (TBSSB) as an innovative anti-mycobacterial drug using as a surrogate model for . We investigated the antimicrobial activity of TBSSB against by plotting growth curves, examined the effect of TBSSB on biofilm formation, observed morphological changes by scanning electron microscopy and transmission electron microscopy, and detected differentially expressed proteins using two-dimensional gel electrophoresis coupled with mass spectrometry.

Role of microtubule-associated protein 6 glycosylated with Gal-(β-1,3)-GalNAc in Parkinson's disease.

Aberrant glycosylation of proteins has major implications for human diseases. To determine whether protein glycosylation contributes to the pathogenesis of Parkinson's disease (PD), a mouse model of PD was established by injection of 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP). Induction of PD-like features was verified by assessing motor impairment and confirming reductions in biological markers, including dopamine, 5-hydroxytryptamine and tyrosine hydroxylase, as well as the aggregation of α-synuclein.

Identification of a novel carboxypeptidase encoded by Rv3627c that plays a potential role in mycobacteria morphology and cell division.

Functionally uncharacterized gene Rv3627c is predicted to encode a carboxypeptidase in the pathogen of Mycobacterium tuberculosis (M. tuberculosis), which remains a major threat to human health. Here, we sought to reveal the function of Rv3627c and to elucidate its effects on mycobacterial growth.

Deficiency of D-alanyl-D-alanine ligase A attenuated cell division and greatly altered the proteome of Mycobacterium smegmatis.

D-Alanyl-D-alanine ligase A (DdlA) catalyses the dimerization of two D-alanines yielding D-alanyl-D-alanine required for mycobacterial peptidoglycan biosynthesis, and is a promising antimycobacterial drug target. To better understand the roles of DdlA in mycobacteria in vivo, we established a cell model in which DdlA expression was specifically downregulated by ddlA antisense RNA by introducing a 380 bp ddlA fragment into pMind followed by transforming the construct into nonpathogenic Mycobacterium smegmatis. The M.

The Biological Properties and Potential Interacting Proteins of d-Alanyl-d-alanine Ligase A from Mycobacterium tuberculosis.

(1) Background: d-alanine-d-alanine ligase (DdlA), an effective target for drug development to combat against (), which threatens human health globally, supplies a substrate of d-alanyl-d-alanine for peptidoglycan crosslinking by catalyzing the dimerization of two d-alanines. To obtain a better understanding of DdlA profiles and develop a colorimetric assay for high-throughput inhibitor screening, we focused on explicating and characterizing -DdlA. (2) Methods and Results: () was expressed in , and the purified -DdlA was identified using (anti)-polyhistidine antibody followed by DdlA activity confirmation by measuring the released orthophosphate via colorimetric assay and the yielded d-alanyl-d-alanine through high performance thin layer chromatography (HP-TLC).

Complete mitochondrial genome and the phylogenetic position of the Borneo leg skate (Rajiformes: Anacanthobatidae).

In this study, the complete mitochondrial genome of the Borneo leg skate (Rajiformes, Anacanthobatidae) was determined. It had circular molecules (16,701 bp), consisting of 37 genes with a typical gene order in vertebrate mitogenome. In the whole mitogenome, there were 28 bp short intergenic and 31 bp overlaps, respectively, located in 12 and 7 gene junctions.

Complete mitochondrial genome and the phylogenetic position of the blackspotted catshark (Carcharhiniformes: Scyliorhinidae).

In this study, the mitogenome of the blackspotted catshark was first determined. This circle molecular was rich in A (30.7%)+T (30.

Complete mitochondrial genome and the phylogenetic position of the White-spotted guitarfish (Rajiformes, Rhinobatidae).

We first determined the complete mitochondrial genome of the White-spotted guitarfish (Rajiformes, Rhinobatidae). The complete mitogenome was 16,804 bp, with a base composition of 32.3% A, 27.

Complete mitochondrial genome and the phylogenetic position of the Jenkins whipray (Myliobatiformes, Dasyatidae).

We determined the complete mitochondrial genome of the Jenkins whipray . The total length of the mitogenome was 17, 670 bp, consisted of 37 genes with typical gene order in vertebrate mitogenome. The nucleotide composition was: 30.

Complete mitochondrial genome and the phylogenetic position of the smalleye pygmy shark (Squaliformes: Dalatiidae).

The complete mitogenome of the smalleye pygmy shark (Squaliformes: Dalatiidae) is presented in this study firstly. It is 16,717 bp with a nucleotide base composition of 30.7% A, 26.

Functionalized collagen scaffold implantation and cAMP administration collectively facilitate spinal cord regeneration.

Unlabelled: Previous studies have demonstrated that several mechanisms, including numerous inhibitory molecules, weak neurotrophic stimulation and deficient intrinsic regenerative responses, collectively contribute to the failure of mature spinal cord axon regeneration. Thus, combinatorial therapies targeting multiple mechanisms have attracted much attention. In the present study, a porous collagen scaffold was used to support neuronal attachment and bridge axonal regeneration.

Functionalized Collagen Scaffold Neutralizing the Myelin-Inhibitory Molecules Promoted Neurites Outgrowth in Vitro and Facilitated Spinal Cord Regeneration in Vivo.

Research has demonstrated that many myelin-associated inhibitory molecules jointly contribute to the failure of adult spinal cord regeneration. Therapies comprehensively targeting the multiple inhibitory nature of the injured spinal cord are being concerned. Here, two collagen-binding proteins, CBD-EphA4LBD and CBD-PlexinB1LBD, were constructed, respectively, to neutralize the axon guidance molecules ephrinB3 and sema4D that inhibit the regeneration of nerve fibers.

Complete mitochondrial genome of the Sharpnose stingray Himantura gerrardi (Myliobatiformes: Dasyatidae).

In this study, we presented the complete mitochondrial genome of the Sharpnose stingray Himantura gerrardi for the first time, which was 17,685 bp in length and contained 13 protein-coding genes, 2 rRNA genes, 22 tRNA genes and a putative control region. The overall nucleotide composition was 30.1% A, 27.

The linear-ordered collagen scaffold-BDNF complex significantly promotes functional recovery after completely transected spinal cord injury in canine.

Spinal cord injury (SCI) is still a worldwide clinical challenge for which there is no viable therapeutic method. We focused on developing combinatorial methods targeting the complex pathological process of SCI. In this study, we implanted linear-ordered collagen scaffold (LOCS) fibers with collagen binding brain-derived neurotrophic factor (BDNF) by tagging a collagen-binding domain (CBD) (LOCS + CBD-BDNF) in completely transected canine SCI with multisystem rehabilitation to validate its potential therapeutic effect through a long-term (38 weeks) observation.

Nerve regeneration and functional recovery by collagen-binding brain-derived neurotrophic factor in an intracerebral hemorrhage model.

Brain-derived neurotrophic factor (BDNF) exerts therapeutic effects following intracerebral hemorrhage (ICH). However, it is difficult to maintain sufficient concentrations in the hemorrhage hemisphere. We demonstrated previously that BDNF fused to a collagen-binding domain (CBD) could bind to collagen in the ventricular ependyma and stimulate cell proliferation in the subventricular zone (SVZ).

Collagen scaffolds modified with CNTF and bFGF promote facial nerve regeneration in minipigs.

Most experiments of peripheral nerve repair after injury have been conducted in the rodent model but the translation of findings from rodent studies to clinical practice is needed partly because the nerve regeneration must occur over much longer distances in humans than in rodents. The reconstruction of long distance nerve injuries still represents a great challenge to surgeons who is engaged in peripheral nerve surgery. Here we used the functional nerve conduit (collagen scaffolds incorporated with neurocytokines CNTF and bFGF) to bridge a 35 mm long facial nerve gap in minipig models.

Methods for detecting transcribed pseudogenes: PCR on regions of high sequence similarity followed by cloning and sequencing.

Identifying pseudogene transcription is problematic in many cases due to the high sequence similarity between pseudogenes and their parental genes. In this chapter, we detail the procedure for the detection of pseudogene transcription using the reverse transcription polymerase chain reaction (RT-PCR) method. The protocol comprises (1) extraction of total RNA, (2) first-strand cDNA synthesis from total RNA, (3) amplification of the cDNA by PCR, and (4) cloning and sequencing of the PCR products.