scholarly journals Comparative transcriptome and proteome profiling of the chlorophyll metabolism pathway in four cultivars of Magnolia officinalis

2018 ◽  
Author(s):  
Qinahua Wu ◽  
Dan Wei ◽  
Yuping Liu ◽  
Chaoxiang Ren ◽  
Qianqian Liu ◽  
...  
Keyword(s):  
Expression Patterns ◽  
Protoporphyrin Ix ◽  
Proteome Analysis ◽  
Chlorophyll Synthesis ◽  
Sichuan Province ◽  
The Other ◽  
Therapeutic Effects ◽  
Uroporphyrinogen Decarboxylase ◽  
Chlorophyll Metabolism ◽  
Magnolia Officinalis

AbstractMagnolia officinalis is an important herb in Chinese medicine that has good therapeutic effects on gastrointestinal motility and helps regulate the spleen and stomach. Magnolia officinalis Rehd.et Wils. (abbreviated as CH)、 Magnolia officinalis Rehd.et Wils. var. biloba’ DaAoye’ (abbreviated as DA)、 Magnolia officinalis Rehd.et Wils. var. biloba‘XiaoAoye’ (abbreviated as XA)、Magnolia officinalis Rehd.et Wils. var. biloba‘Liuye’ (abbreviated as LY) are the four main cultivars in Dujiangyan city, Sichuan province in China, and DA has better medicinal effects and higher levels of green pigment than the other three cultivars. Proteomics research not only provides new insights into protein expression patterns but also enables the identification of many attractive candidates for further research on the effects of pesticides on chlorophyll metabolism in Magnolia officinalis. Transcriptome and proteome analysis of the four main Magnolia officinalis cultivars in Sichuan province in China identified 125,100 unigenes and 6,012 proteins. Proteomic data and parallel reaction monitoring (PRM) revealed that porphobilinogen deaminase, uroporphyrinogen decarboxylase, oxygen-dependent coproporphyrinogen-III oxidase, protoporphyrinogen oxidase 1, magnesium protoporphyrin IX methyltransferase, magnesium-protoporphyrin IX monomethyl ester oxidative cyclase, and protochlorophyllide reductase-like protein, all of which are involved in chlorophyll synthesis, have higher expression levels in DA than in the other three cultivars. This finding is consistent with the observation that DA has the highest concentration of chlorophyll (37.56 mg/g Fw) of the four cultivars. This research is beneficial to the understanding of the differences in the growth of the four cultivars on the molecular level.

scholarly journals Dental pulp stem cells overexpressing hepatocyte growth factor facilitate the repair of DSS-induced ulcerative colitis

2021 ◽  
Vol 12 (1) ◽  
Author(s):  
Ning Li ◽  
Yichi Zhang ◽  
Narayan Nepal ◽  
Guoqing Li ◽  
Ningning Yang ◽  
...  
Keyword(s):  
Oxidative Stress ◽  
Ulcerative Colitis ◽  
Body Weight ◽  
Stem Cell ◽  
Dental Pulp ◽  
The Other ◽  
Therapeutic Effects ◽  
Colon Injury ◽  
Treatment Groups ◽  
Hepatocyte Growth

Abstract Background Ulcerative colitis (UC) is a chronic and recurrent disease without satisfactory treatment strategies. Dental pulp stem cell (DPSC) transplantation has been proposed as a potential therapy for UC. This study aimed to investigate the therapeutic effects of the rat hepatocyte growth factor (HGF) gene transduced into DPSCs for UC. Methods The therapeutic effects of HGF-DPSCs transplanted intravenously into a rat model of UC induced by 5% dextran sulphate sodium (DSS) were compared with the other treatment groups (LV-HGF group, DPSCs group and GFP-DPSCs group). Immunofluorescence and immunohistochemistry were used to observe the localization and proliferation of HGF-DPSCs at the site of colon injury. The expression levels of inflammatory factors were detected by real-time quantitative PCR (RT-PCR) and western blotting. The oxidative stress markers were detected by ELISA. DAI scores and body weight changes were used to macroscopically evaluate the treatment of rats in each group. Results Immunofluorescence and immunohistochemistry assays showed that HGF-DPSCs homed to colon injury sites and colocalized with intestinal stem cell (ISC) markers (Bmi1, Musashi1 and Sox9) and significantly promoted protein expression (Bmi1, Musashi1, Sox9 and PCNA). Anti-inflammatory cytokine (TGF-β and IL-10) expression was the highest in the HGF-DPSCs group compared with the other treatment groups, while the expression of pro-inflammatory cytokines (TNF-α and INF-γ) was the lowest. Additionally, the oxidative stress response results showed that malondialdehyde (MDA) and myeloperoxidase (MPO) expression decreased while superoxide dismutase (SOD) expression increased, especially in the HGF-DPSCs group. The DAI scores showed a downward trend with time in the five treatment groups, whereas body weight increased, and the changes were most prominent in the HGF-DPSCs group. Conclusions The study indicated that HGF-DPSCs can alleviate injuries to the intestinal mucosa by transdifferentiating into ISC-like cells, promoting ISC-like cell proliferation, suppressing inflammatory responses and reducing oxidative stress damage, which provides new ideas for the clinical treatment of UC.

Cloning and mapping of human PKIB and PKIG, and comparison of tissue expression patterns of three members of the protein kinase inhibitor family, including PKIA

2000 ◽  
Vol 349 (2) ◽  
pp. 403-407 ◽  
Author(s):  
Lihua ZHENG ◽  
Long YU ◽  
Qiang TU ◽  
Min ZHANG ◽  
Hua HE ◽  
...  
Keyword(s):  
Skeletal Muscle ◽  
Protein Kinase ◽  
Nuclear Export ◽  
Kinase Inhibitor ◽  
Expression Patterns ◽  
Nuclear Export Signal ◽  
Tissue Expression ◽  
Protein Kinase Inhibitor ◽  
The Other ◽  
Dependent Protein

Two novel members of the human cAMP-dependent protein kinase inhibitor (PKI) gene family, PKIB and PKIG, were cloned. The deduced proteins showed 70% and 90% identity with mouse PKIβ and PKIγ respectively. Both the already identified pseudosubstrate site and leucine-rich nuclear export signal motifs were defined from the 11 PKIs of different species. The PKIB and PKIG genes were mapped respectively to chromosome 6q21-22.1, using a radiation hybrid GB4 panel, and to chromosome 20q13.12-13.13, using a Stanford G3 panel. Northern-blot analysis of three PKI isoforms, including the PKIA identified previously, revealed significant differences in their expression patterns. PKIB had two transcripts of 1.9 kb and 1.4 kb. The former transcript was abundant in both placenta and brain and the latter was expressed most abundantly in placenta, highly in brain, heart, liver, pancreas, moderately in kidney, skeletal muscle and colon, and very little in the other eight tissues tested. PKIG was widely expressed as a 1.5-kb transcript with the highest level in heart, hardly detectable in thymus and peripheral blood leucocytes and was moderately expressed in the other tissues, with slightly different levels. However, PKIA was specifically expressed as two transcripts of 3.3 kb and 1.5 kb in heart and skeletal muscle. The distinct expression patterns of the three PKIs suggest that their roles in various tissues are probably different.

scholarly journals Transcriptome and Proteome Research in Veterinary Science: What Is Possible and What Questions Can Be Asked?

2012 ◽  
Vol 2012 ◽  
pp. 1-14 ◽  
Author(s):  
Robert Klopfleisch ◽  
Achim D. Gruber
Keyword(s):  
Sequential Analysis ◽  
Biomarker Discovery ◽  
Expression Patterns ◽  
Proteome Analysis ◽  
Shotgun Proteomics ◽  
Complete Analysis ◽  
Biological Research ◽  
Veterinary Research ◽  
Gene Sets ◽  
2D Gel

In recent years several technologies for the complete analysis of the transcriptome and proteome have reached a technological level which allows their routine application as scientific tools. The principle of these methods is the identification and quantification of up to ten thousands of RNA and proteins species in a tissue, in contrast to the sequential analysis of conventional methods such as PCR and Western blotting. Due to their technical progress transcriptome and proteome analyses are becoming increasingly relevant in all fields of biological research. They are mainly used for the explorative identification of disease associated complex gene expression patterns and thereby set the stage for hypothesis-driven studies. This review gives an overview on the methods currently available for transcriptome analysis, that is, microarrays, Ref-Seq, quantitative PCR arrays and discusses their potentials and limitations. Second, the most powerful current approaches to proteome analysis are introduced, that is, 2D-gel electrophoresis, shotgun proteomics, MudPIT and the diverse technological concepts are reviewed. Finally, experimental strategies for biomarker discovery, experimental settings for the identification of prognostic gene sets and explorative versus hypothesis driven approaches for the elucidation of diseases associated genes and molecular pathways are described and their potential for studies in veterinary research is highlighted.

An evolutionary and developmental approach toward understanding of growth variations of two closely related dwarfism of Galium verum

Biologia ◽  
2014 ◽  
Vol 69 (12) ◽  
Author(s):  
Keum Jeong ◽  
Jae-Hong Pak ◽  
Jeong Kim
Keyword(s):  
Cell Proliferation ◽  
Genetic Factors ◽  
Expression Patterns ◽  
Mrna Differential Display ◽  
Growth Patterns ◽  
The Other ◽  
Differential Growth ◽  
Developmental Approach ◽  
Differential Display Method ◽  
Low Activity

AbstractGalium L. is one of the largest and most widespread genus of Rubiaceae, consisting of more than 650 species worldwide. Galium verum var. asiaticum (G. verum a.) is a perennial herbaceous and widely distributed in in Korea peninsula. On the other hand, Galium verum var. asiaticum for. pusillum (G. verum a.p.) is endemic to Korea, inhabiting only on high land of Mt. Halla of Jeju. G. verum a.p. appears to be a dwarfism of G. verum a. We wondered what physiological, environmental, or genetic factors rendered those two taxa morphologically differentiated. We found that G. verum a.p. shows a low activity of the cell proliferation and was not associated with responsiveness contents of auxin and gibberellins. In order to search for genetic factors involved, we carried out an mRNA differential display method using the ACPs, and isolated several different expression genes between the two taxa. We chose one of those genes, which encoded RADIALIS-like proteins: GvaRADL1 from G. verum a. and GvapRADL1 from G. verum a.p. We discuss the relevancy of the genetic variations in regard to the differential expression patterns of those genes and the differential growth patterns of the two variants.

Single Cell Profiling Reveals Unique CXCL13 Positive T Cell Subsets in the Tumor Microenvironment of Lymphocyte Rich Classic Hodgkin Lymphoma

Blood ◽  
2020 ◽  
Vol 136 (Supplement 1) ◽  
pp. 32-33
Author(s):  
Tomohiro Aoki ◽  
Lauren C. Chong ◽  
Katsuyoshi Takata ◽  
Katy Milne ◽  
Elizabeth Chavez ◽  
...  
Keyword(s):  
T Cells ◽  
T Cell ◽  
B Cells ◽  
Single Cell ◽  
Immune Cells ◽  
Research Funding ◽  
Immune Cell ◽  
Expression Patterns ◽  
The Other ◽  
Tfh Cells

Introduction: Classic Hodgkin lymphoma (CHL) features a unique crosstalk between malignant cells and different types of normal immune cells in the tumor-microenvironment (TME). On the basis of histomorphologic and immunophenotypic features of the malignant Hodgkin and Reed-Sternberg (HRS) cells and infiltrating immune cells, four histological subtypes of CHL are recognized: Nodular sclerosing (NS), Mixed cellularity, Lymphocyte-rich (LR) and Lymphocyte-depleted CHL. Recently, our group described the high abundance of various types of immunosuppressive CD4+ T cells including LAG3+ and/or CTLA4+ cells in the TME of CHL using single cell RNA sequencing (scRNAseq). However, the TME of LR-CHL has not been well characterized due to the rarity of the disease. In this study, we aimed at characterizing the immune cell profile of LR-CHL at single cell resolution. METHODS: We performed scRNAseq on cell suspensions collected from lymph nodes of 28 primary CHL patients, including 11 NS, 9 MC and 8 LR samples, with 5 reactive lymph nodes (RLN) serving as normal controls. We merged the expression data from all cells (CHL and RLN) and performed batch correction and normalization. We also performed single- and multi-color immunohistochemistry (IHC) on tissue microarray (TMA) slides from the same patients. In addition, an independent validation cohort of 31 pre-treatment LR-CHL samples assembled on a TMA, were also evaluated by IHC. Results: A total of 23 phenotypic cell clusters were identified using unsupervised clustering (PhenoGraph). We assigned each cluster to a cell type based on the expression of genes described in published transcriptome data of sorted immune cells and known canonical markers. While most immune cell phenotypes were present in all pathological subtypes, we observed a lower abundance of regulatory T cells (Tregs) in LR-CHL in comparison to the other CHL subtypes. Conversely, we found that B cells were enriched in LR-CHL when compared to the other subtypes and specifically, all four naïve B-cell clusters were quantitatively dominated by cells derived from the LR-CHL samples. T follicular helper (TFH) cells support antibody response and differentiation of B cells. Our data show the preferential enrichment of TFH in LR-CHL as compared to other CHL subtypes, but TFH cells were still less frequent compared to RLN. Of note, Chemokine C-X-C motif ligand 13 (CXCL13) was identified as the most up-regulated gene in LR compared to RLN. CXCL13, which is a ligand of C-X-C motif receptor 5 (CXCR5) is well known as a B-cell attractant via the CXCR5-CXCL13 axis. Analyzing co-expression patterns on the single cell level revealed that the majority of CXCL13+ T cells co-expressed PD-1 and ICOS, which is known as a universal TFH marker, but co-expression of CXCR5, another common TFH marker, was variable. Notably, classical TFH cells co-expressing CXCR5 and PD-1 were significantly enriched in RLN, whereas PD-1+ CXCL13+ CXCR5- CD4+ T cells were significantly enriched in LR-CHL. These co-expression patterns were validated using flow cytometry. Moreover, the expression of CXCR5 on naïve B cells in the TME was increased in LR-CHL compared to the other CHL subtypes We next sought to understand the spatial relationship between CXCL13+ T cells and malignant HRS cells. IHC of all cases revealed that CXCL13+ T cells were significantly enriched in the LR-CHL TME compared to other subtypes of CHL, and 46% of the LR-CHL cases showed CXCL13+ T cell rosettes closely surrounding HRS cells. Since PD-1+ T cell rosettes are known as a specific feature of LR-CHL, we confirmed co-expression of PD-1 in the rosetting cells by IHC in these cases. Conclusions: Our results reveal a unique TME composition in LR-CHL. LR-CHL seems to be distinctly characterized among the CHL subtypes by enrichment of CXCR5+ naïve B cells and CD4+ CXCL13+ PD-1+ T cells, indicating the importance of the CXCR5-CXCL13 axis in the pathogenesis of LR-CHL. Figure Disclosures Savage: BeiGene: Other: Steering Committee; Merck, BMS, Seattle Genetics, Gilead, AstraZeneca, AbbVie: Honoraria; Roche (institutional): Research Funding; Merck, BMS, Seattle Genetics, Gilead, AstraZeneca, AbbVie, Servier: Consultancy. Scott:Janssen: Consultancy, Research Funding; Celgene: Consultancy; NanoString: Patents & Royalties: Named inventor on a patent licensed to NanoString, Research Funding; NIH: Consultancy, Other: Co-inventor on a patent related to the MCL35 assay filed at the National Institutes of Health, United States of America.; Roche/Genentech: Research Funding; Abbvie: Consultancy; AstraZeneca: Consultancy. Steidl:AbbVie: Consultancy; Roche: Consultancy; Curis Inc: Consultancy; Juno Therapeutics: Consultancy; Bayer: Consultancy; Seattle Genetics: Consultancy; Bristol-Myers Squibb: Research Funding.

Cocaine in the Treatment of Mental Disorders

1887 ◽  
Vol 33 (142) ◽  
pp. 230-238 ◽  
Author(s):  
C. Heimann
Keyword(s):  
Mental Disorders ◽  
Single Case ◽  
The Other ◽  
Human Organism ◽  
Therapeutic Effects ◽  
Permanent Cure ◽  
The One

Certain therapeutic effects upon the human organism ascribed to cocaine,∗ occasioned me to make use of the drug in suitable cases of psychosis and psycho-neurosis. Stimulant action and exhilaration (Euphoria) on the one hand, and on the other depression of undue sensitiveness, these were the effects I looked for from the alkaloid. Unfortunately, I am able to record scarcely a single case of certain and permanent cure following the use of the drug.

Measurement of ferrochelatase activity using a novel assay suggests that plastids are the major site of haem biosynthesis in both photosynthetic and non-photosynthetic cells of pea (Pisum sativum L.)

2002 ◽  
Vol 362 (2) ◽  
pp. 423-432 ◽  
Author(s):  
Johanna E. CORNAH ◽  
Jennifer M. ROPER ◽  
Davinder Pal SINGH ◽  
Alison G. SMITH
Keyword(s):  
Ferrous Iron ◽  
Specific Activity ◽  
Higher Plants ◽  
Protoporphyrin Ix ◽  
Chlorophyll Synthesis ◽  
Marker Enzyme ◽  
Hexane Extraction ◽  
Higher Plant ◽  
Major Site ◽  
Haem Biosynthesis

Ferrochelatase is the terminal enzyme of haem biosynthesis, catalysing the insertion of ferrous iron into the macrocycle of protoporphyrin IX, the last common intermediate of haem and chlorophyll synthesis. Its activity has been reported in both plastids and mitochondria of higher plants, but the relative amounts of the enzyme in the two organelles are unknown. Ferrochelatase is difficult to assay since ferrous iron requires strict anaerobic conditions to prevent oxidation, and in photosynthetic tissues chlorophyll interferes with the quantification of the product. Accordingly, we developed a sensitive fluorimetric assay for ferrochelatase that employs Co2+ and deuteroporphyrin in place of the natural substrates, and measures the decrease in deuteroporphyrin fluorescence. A hexane-extraction step to remove chlorophyll is included for green tissue. The assay is linear over a range of chloroplast protein concentrations, with an average specific activity of 0.68nmol·min−1·mg of protein−1, the highest yet reported. The corresponding value for mitochondria is 0.19nmol·min−1·mg of protein−1. The enzyme is inhibited by N-methylprotoporphyrin, with an estimated IC50 value of ≈ 1nM. Using this assay we have quantified ferrochelatase activity in plastids and mitochondria from green pea leaves, etiolated pea leaves and pea roots to determine the relative amounts in the two organelles. We found that, in all three tissues, greater than 90% of the activity was associated with plastids, but ferrochelatase was reproducibly detected in mitochondria, at levels greater than the contaminating plastid marker enzyme, and was latent. Our results indicate that plastids are the major site of haem biosynthesis in higher plant cells, but that mitochondria also have the capacity for haem production.

scholarly journals Cell-Penetrating Peptide and siRNA-Mediated Therapeutic Effects on Endometriosis and Cancer In Vitro Models

Pharmaceutics ◽  
2021 ◽  
Vol 13 (10) ◽  
pp. 1618
Author(s):  
Kristina Kiisholts ◽  
Kaido Kurrikoff ◽  
Piret Arukuusk ◽  
Ly Porosk ◽  
Maire Peters ◽  
...  
Keyword(s):  
Gene Therapy ◽  
Cellular Proliferation ◽  
Expression Patterns ◽  
In Vitro Models ◽  
Cell Penetrating Peptides ◽  
Biologically Active ◽  
Therapeutic Effects ◽  
Cell Penetrating ◽  
Proliferation And Invasion

Gene therapy is a powerful tool for the development of new treatment strategies for various conditions, by aiming to transport biologically active nucleic acids into diseased cells. To achieve that goal, we used highly potential delivery vectors, cell-penetrating peptides (CPPs), as oligonucleotide carriers for the development of a therapeutic approach for endometriosis and cancer. Despite marked differences, both of these conditions still exhibit similarities, like excessive, uncoordinated, and autonomous cellular proliferation and invasion, accompanied by overlapping gene expression patterns. Thus, in the current study, we investigated the therapeutic effects of CPP and siRNA nanoparticles using in vitro models of benign endometriosis and malignant glioblastoma. We demonstrated that CPPs PepFect6 and NickFect70 are highly effective in transfecting cell lines, primary cell cultures, and three-dimensional spheroids. CPP nanoparticles are capable of inducing siRNA-specific knockdown of therapeutic genes, ribonucleotide reductase subunit M2 (RRM2), and vascular endothelial growth factor (VEGF), which results in the reduction of in vitro cellular proliferation, invasion, and migration. In addition, we proved that it is possible to achieve synergistic suppression of endometriosis cellular proliferation and invasion by combining gene therapy and hormonal treatment approaches by co-administering CPP/siRNA nanoparticles together with the endometriosis-drug danazol. We suggest a novel target, RRM2, for endometriosis therapy and as a proof-of-concept, we propose a CPP-mediated gene therapy approach for endometriosis and cancer.

Quality of Glucosamine Products: Is it a Potential Reason for Inconsistent Clinical Outcomes in Osteoarthritis?

2020 ◽  
Author(s):  
Chhavi Asthana ◽  
Gregory M Peterson ◽  
Madhur D Shastri ◽  
Rahul P Patel
Keyword(s):  
United States ◽  
Clinical Studies ◽  
The United States ◽  
The Other ◽  
Quality Analysis ◽  
Therapeutic Effects ◽  
Clinical Effects ◽  
United States Pharmacopoeia ◽  
Potential Reason

Abstract Background Clinical studies have reported inconsistent outcomes of glucosamine therapy in osteoarthritis patients. One possible reason could be the use of glucosamine products of varying quality. Objective Hence, this study aimed to assess the quality of glucosamine products marketed in Australia and India. This is the first study to investigate both the content and dissolution profiles of glucosamine products. Method The content and dissolution analysis of Australian (n = 25 brands) and Indian (n = 21 brands) glucosamine products was performed according to the criteria specified in the United States Pharmacopoeia (USP). Results The quality analysis revealed that 16% and 18% of Australian brands, as well as 24% and 19% of Indian brands, did not fulfil the USP content and dissolution criteria, respectively. In approximately half of these cases, the glucosamine content was only slightly below (<3%) that specified by the USP and dissolution was achieved within 15 min after the duration specified by the USP. Conclusions The majority of the brands did meet both the content and dissolution analysis criteria of the USP. The extent of deviation from the specified criteria for the other brands was probably insufficient to account for the significant variability in clinical effects. Hence, the study proposed that inter-patient pharmacokinetic variations in glucosamine could be another potential reason for inconsistent therapeutic effects. Highlights

Interactions entre le cycloheximide et la 6-benzylaminopurine au cours du verdissement de cotylédons de concombre

1980 ◽  
Vol 58 (9) ◽  
pp. 1101-1110 ◽  
Author(s):  
J. Roussaux ◽  
M. Hoffelt ◽  
N. Farineau
Keyword(s):  
Cucumis Sativus ◽  
Dark Period ◽  
Chlorophyll Synthesis ◽  
The Other ◽  
Plastid Differentiation ◽  
Cucumis Sativus L ◽  
Modes Of Action ◽  
Plastid Ultrastructure ◽  
Ultrastructural Effect

Etiolated cotyledons of cucumber (Cucumis sativus L.) were isolated and incubated for 15 h in the dark and 48 h in the light in the presence of 10−6 g/mL 6-benzylaminopurine (6-BAP) or 3.10−6 and 10−5 g/mL cycloheximide (CHX). Cotyledons were permanently maintained on these compounds or transferred from one medium to the other after the dark period or after 6 h of the light. 6-BAP stimulates both chlorophyll synthesis and plastid differentiation. Chlorophyll synthesis is more affected than differentiation in the presence of CHX. Only some thylakoids are present in plastids treated permanently with CHX at 10−5 g/mL. At 3.10−6 g/mL, CHX inhibits the development of the stroma thylakoids and the granal stacks. The effect of CHX is particularly important during the 1st h of light; it is partially reversible. During the treatment with 6-BAP followed by CHX (or conversely), the ultrastructural effect of 10−5 g/mL CHX is removed if 6-BAP is given before the inhibitor. After 48 h the plastid ultrastructure depends particularly on the compound applied during the 1st h of light. 6-BAP and CHX are not competitive during chlorophyllogenesis. 6-BAP increases and CHX decreases the incorporation of 32P into cytoplasmic and etioplastic rRNA during the dark. During the light, 6-BAP increases the incorporation in chloroplastic rRNA only and CHX decreases the incorporation in cytoplasmic rRNA only. The modes of action of CHX and its relations with 6-BAP are discussed.

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