Transfecting Taxus � media Protoplasts to Study Transcription Factors BIS2 and TSAR2 as Activators of Taxane-Related Genes

Abstract Taxane diterpenes are secondary metabolites with an important pharmacological role in the treatment of cancer. Taxus spp. biofactories have been used for taxane production, but the lack of knowledge about the taxane biosynthetic pathway and its molecular regulation hinders their optimal function. The difficulties in introducing foreign genes in Taxus spp. genomes hinder the study of the molecular mechanisms involved in taxane production, and a new approach is required to overcome them. In this study, a reliable, simple and fast method to obtain Taxus � media protoplasts was developed, allowing their manipulation in downstream assays for the study of physiological changes in Taxus spp. cells. Using this method, Taxus protoplasts were transiently transfected for the first time, corroborating their suitability for transfection assays and the study of specific physiological responses. The two assayed transcription factors (BIS2 and TSAR2) had a positive effect on the expression of several taxane-related genes, suggesting their potential use for the improvement of taxane yields. Furthermore, the results indicate that the developed method is suitable for obtaining T. � media protoplasts for transfection with the aim of unraveling regulatory mechanisms in taxane production.

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The Role of Transcription Factors in Gonad Development and Sex Differentiation of a Teleost Model Fish—Guppy (Poecilia reticulata)

Animals ◽

10.3390/ani10122401 ◽

2020 ◽

Vol 10 (12) ◽

pp. 2401

Author(s):

Maciej Kamaszewski ◽

Marek Skrobisz ◽

Maciej Wójcik ◽

Kacper Kawalski ◽

Adrian Szczepański ◽

...

Keyword(s):

Transcription Factors ◽

Poecilia Reticulata ◽

Molecular Mechanisms ◽

Histological Analysis ◽

Sex Differentiation ◽

Gonad Development ◽

Toxicological Studies ◽

Development And Differentiation ◽

First Time ◽

Germinal Cells

The guppy (Poecilia reticulata) is one of the world’s most popular ornamental fish. Due to lecithotrophic viviparous, it is commonly used in toxicological studies and environmental monitoring. This study aimed to investigate the molecular mechanisms of gonad development and differentiation during guppy ontogenesis. The study mainly focused on the role and localization of potential specific sex markers and transcription factors: Sox9, Dmrt1, Erβ. For histological analysis, guppies of both sexes were collected at 1, 60, and 360 dph (day post-hatching). The gonads morphology and immunohistochemistry detection of mentioned markers localization were performed. The expression of Sox9 protein was compared between sexes. Histological analysis revelated all types of male germinal cells in 60 dph guppy’s testes. Maturated oocytes were visible in the ovaries of 360 dph fish. The Sox9 expression varied in spermatocytes and spermatids, from cellular to nuclear localization, and was higher in ovaries. Dmrt1 was detected in all testes groups and 360 dph ovaries. The Erβ was observed in both sexes at 60 and 360 dph. For the first time, the localization of transcription factors in guppy during ontogenesis was traced. The Sox9 designation as a factor regulating the development of germinal cells in adult guppies may facilitate the analysis of xenobiotics’ influence on fish’s reproductive system.

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TAMM HORSFALL PROTEIN: THE MEN BEHIND THE EPONYM

Nephrology (Saint-Petersburg) ◽

10.24884/1561-6274-2019-23-2-117-119 ◽

2019 ◽

Vol 23 (2) ◽

pp. 117-119 ◽

Cited By ~ 1

Author(s):

D. N. Paskalev ◽

B. T. Galunska ◽

D. Petkova-Valkova

Keyword(s):

Viral Replication ◽

Research Team ◽

Molecular Mechanisms ◽

Thick Ascending Limb ◽

Rockefeller Institute ◽

The Usa ◽

Natural Inhibitor ◽

Simplex Virus ◽

First Time ◽

New Protein

Tamm–Horsfall Protein (uromodulin) is named after Igor Tamm and Franc Horsfall Jr who described it for the first time in 1952. It is a glycoprotein, secreted by the cells in the thick ascending limb of the loop of Henle. This protein will perform a number of important pathophysiological functions, including protection against uroinfections, especially caused by E. Сoli, and protection against formation of calcium concernments in the kidney. Igor Tamm (1922-1995) is an outstanding cytologist, virologist and biochemist. He is one of the pioneers in the study of viral replication. He was born in Estonia and died in the USA. In 1964 he was elected for a professorship in Rockefeller Institute for Medical Research, where has been working continuously. Since 1959, he became a head of the virology lab established by his mentor and co-author Franc Horsfall. In the course of studies on the natural inhibitor of viral replication, Tamm and Horsfall isolated and characterized biochemically a new protein named after their names. Franc Lappin Horsfall Jr (1906-1971) was a well-known clinician and virologist with remarkable achievements in internal medicine. He was born and died in the USA. He worked in the Rockefeller Hospital from 1934 to 1960, then in the Center for Cancer Research at the Sloan-Kettering Institute. Here he was a leader of a research team studying the molecular mechanisms of immunity, the effects of chemotherapy with benzimidazole compounds (together with I. Tamm), coxsackie viruses, herpes simplex virus, etc.

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Combining QTL Mapping and Gene Expression Analysis to Elucidate the Genetic Control of ‘Crumbly’ Fruit in Red Raspberry (Rubus idaeus L.)

Agronomy ◽

10.3390/agronomy11040794 ◽

2021 ◽

Vol 11 (4) ◽

pp. 794

Author(s):

Luca M. Scolari ◽

Robert D. Hancock ◽

Pete E. Hedley ◽

Jenny Morris ◽

Kay Smith ◽

...

Keyword(s):

Genetic Control ◽

Developmental Disorder ◽

Molecular Mechanisms ◽

Developmental Stages ◽

Microarray Experiment ◽

Rubus Idaeus ◽

Red Raspberry ◽

Genotype By Sequencing ◽

First Time ◽

Selection Of

‘Crumbly’ fruit is a developmental disorder in raspberry that results in malformed and unsaleable fruits. For the first time, we define two distinct crumbly phenotypes as part of this work. A consistent crumbly fruit phenotype affecting the majority of fruits every season, which we refer to as crumbly fruit disorder (CFD) and a second phenotype where symptoms vary across seasons as malformed fruit disorder (MFD). Here, segregation of crumbly fruit of the MFD phenotype was examined in a full-sib family and three QTL (Quantitative Trait Loci) were identified on a high density GbS (Genotype by Sequencing) linkage map. This included a new QTL and more accurate location of two previously identified QTLs. A microarray experiment using normal and crumbly fruit at three different developmental stages identified several genes that were differentially expressed between the crumbly and non-crumbly phenotypes within the three QTL. Analysis of gene function highlighted the importance of processes that compromise ovule fertilization as triggers of crumbly fruit. These candidate genes provided insights regarding the molecular mechanisms involved in the genetic control of crumbly fruit in red raspberry. This study will contribute to new breeding strategies and diagnostics through the selection of molecular markers associated with the crumbly trait.

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Reactive Dual Magnetron Sputtering: A Fast Method for Preparing Stoichiometric Microcrystalline ZnWO4 Thin Films

Surfaces ◽

10.3390/surfaces4020013 ◽

2021 ◽

Vol 4 (2) ◽

pp. 106-114

Author(s):

Yannick Hermans ◽

Faraz Mehmood ◽

Kerstin Lakus-Wollny ◽

Jan P. Hofmann ◽

Thomas Mayer ◽

...

Keyword(s):

Thin Films ◽

Magnetron Sputtering ◽

Photoelectron Spectroscopy ◽

Fast Method ◽

X Ray Diffraction ◽

X Ray ◽

Glass Substrates ◽

Post Annealing ◽

Rf Signal ◽

First Time

Thin films of ZnWO4, a promising photocatalytic and scintillator material, were deposited for the first time using a reactive dual magnetron sputtering procedure. A ZnO target was operated using an RF signal, and a W target was operated using a DC signal. The power on the ZnO target was changed so that it would match the sputtering rate of the W target operated at 25 W. The effects of the process parameters were characterized using optical spectroscopy, X-ray diffraction, and scanning electron microscopy, including energy dispersive X-ray spectroscopy as well as X-ray photoelectron spectroscopy. It was found that stoichiometric microcrystalline ZnWO4 thin films could be obtained, by operating the ZnO target during the sputtering procedure at a power of 55 W and by post-annealing the resulting thin films for at least 10 h at 600 °C. As FTO coated glass substrates were used, annealing led as well to the incorporation of Na, resulting in n+ doped ZnWO4 thin films.

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Analysis of a Preliminary microRNA Expression Signature in a Human Telangiectatic Osteogenic Sarcoma Cancer Cell Line

International Journal of Molecular Sciences ◽

10.3390/ijms22031163 ◽

2021 ◽

Vol 22 (3) ◽

pp. 1163

Author(s):

Gaia Palmini ◽

Cecilia Romagnoli ◽

Simone Donati ◽

Roberto Zonefrati ◽

Gianna Galli ◽

...

Keyword(s):

Cell Line ◽

Molecular Mechanisms ◽

Osteogenic Sarcoma ◽

Cancer Cell Line ◽

Microscopic Features ◽

In Vitro Establishment ◽

Profile Characteristic ◽

First Time

Telangiectatic osteosarcoma (TOS) is an aggressive variant of osteosarcoma (OS) with distinctive radiographic, gross, microscopic features, and prognostic implications. Despite several studies on OS, we are still far from understanding the molecular mechanisms of TOS. In recent years, many studies have demonstrated not only that microRNAs (miRNAs) are involved in OS tumorigenesis, development, and metastasis, but also that the presence in high-grade types of OS of cancer stem cells (CSCs) plays an important role in tumor progression. Despite these findings, nothing has been described previously about the expression of miRNAs and the presence of CSCs in human TOS. Therefore, we have isolated/characterized a putative CSC cell line from human TOS (TOS-CSCs) and evaluated the expression levels of several miRNAs in TOS-CSCs using real-time quantitative assays. We show, for the first time, the existence of CSCs in human TOS, highlighting the in vitro establishment of this unique stabilized cell line and an identification of a preliminary expression of the miRNA profile, characteristic of TOS-CSCs. These findings represent an important step in the study of the biology of one of the most aggressive variants of OS and the role of miRNAs in TOS-CSC behavior.

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Wheat Bread with Grape Seeds Micropowder: Impact on Dough Rheology and Bread Properties

Applied Rheology ◽

10.1515/arh-2020-0112 ◽

2020 ◽

Vol 30 (1) ◽

pp. 138-150

Author(s):

Veronika Valková ◽

Hana Ďúranová ◽

Jana Štefániková ◽

Michal Miškeje ◽

Marián Tokár ◽

...

Keyword(s):

Wheat Flour ◽

Dough Rheology ◽

Wheat Bread ◽

Grape Seeds ◽

Rheological Analysis ◽

First Time ◽

Positive Effect ◽

Bread Properties ◽

Electronic Eye ◽

Different Levels

AbstractThe current study was designed to enhance the functionality of white bread by replacement of wheat flour with different levels (1%, 2%, 5%, and 8%) of grape seeds micropowder (GSMP) with nanosized particles (10 µm). Chemical composition of GSMP, volume and sensory attributes, evaluated with the panel of evaluators and an electronic nose (e-nose) and an electronic eye (e-eye) were investigated in the tested breads. It has been found out that GSMP contained appreciable amounts of flavonoids including catechin, epicatechin, gallic acid and minerals especially, Ca, K and Mg. The data from rheological analysis showed that the addition of GSMP (mainly at 5% and 8% levels) to the wheat flour had a positive effect on dough manifesting with rheology by increased dough stability. The volume of the experimental breads (above 1% concentration) was demonstrably declined (P < 0.0001) in comparison with the control bread. Sensory rating revealed that the bread fortified with 1% GSMP was judged by the consumer panelists as the most acceptable with the highest scores for all quality attributes which was also confirmed by the data of e-nose and e-eye. Our results suggest for the first time that 1% GSMP addition appears to be a promising functional ingredient to improve bread with required qualitative and sensory properties.

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High Energy Intake Induced Overexpression of Transcription Factors and Its Regulatory Genes Involved in Acceleration of Hepatic Lipogenesis: A Rat Model for Type 2 Diabetes

Biomedicines ◽

10.3390/biomedicines7040076 ◽

2019 ◽

Vol 7 (4) ◽

pp. 76 ◽

Cited By ~ 3

Author(s):

Suresh P. Khadke ◽

Aniket A. Kuvalekar ◽

Abhay M. Harsulkar ◽

Nitin Mantri

Keyword(s):

Type 2 Diabetes ◽

Transcription Factors ◽

Glucose Tolerance ◽

Molecular Mechanisms ◽

Target Genes ◽

High Energy ◽

Tolerance Test ◽

Diabetic Control ◽

Ppar Γ

Type 2 diabetes mellitus (T2DM) is a metabolic disorder characterized by impaired insulin action and its secretion. The objectives of the present study were to establish an economical and efficient animal model, mimicking pathophysiology of human T2DM to understand probable molecular mechanisms in context with lipid metabolism. In the present study, male Wistar rats were randomly divided into three groups. Animals were fed with high fat diet (HFD) except healthy control (HC) for 12 weeks. After eight weeks, intra peritoneal glucose tolerance test was performed. After confirmation of glucose intolerance, diabetic control (DC) group was injected with streptozotocin (STZ) (35 mg/kg b.w., i.p.). HFD fed rats showed increase (p ≤ 0.001) in glucose tolerance and HOMA-IR as compared to HC. Diabetes rats showed abnormal (p ≤ 0.001) lipid profile as compared to HC. The hepatocyte expression of transcription factors SREBP-1c and NFκβ, and their target genes were found to be upregulated, while PPAR-γ, CPT1A and FABP expressions were downregulated as compared to the HC. A number of animal models have been raised for studying T2DM, but the study has been restricted to only the biochemical level. The model is validated at biochemical, molecular and histopathological levels, which can be used for screening new therapeutics for the effective management of T2DM.

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A multi-manned assembly line balancing problem with classified teams: a new approach

Assembly Automation ◽

10.1108/aa-04-2015-035 ◽

2016 ◽

Vol 36 (1) ◽

pp. 51-59 ◽

Cited By ~ 15

Author(s):

Hamid Yilmaz ◽

Mustafa Yilmaz

Keyword(s):

Design Methodology ◽

Assembly Line ◽

Assembly Line Balancing ◽

Line Balancing ◽

Test Problems ◽

New Approach ◽

Content Type ◽

Line Test ◽

Assembly Line Balancing Problem ◽

First Time

Purpose – Within team-oriented approaches, tasks are assigned to teams before being assigned to workstations as a reality of industry. So it becomes clear, which workers assemble which tasks. Design/methodology/approach – Team numbers of the assembly line can increase with the number of tasks, but at the same time, due to physical situations of the stations, there will be limitations of maximum working team numbers in a station. For this purpose, heuristic assembly line balancing (ALB) procedure is used and mathematical model is developed for the problem. Findings – Well-known assembly line test problems widely used in the literature are solved to indicate the effectiveness and applicability of the proposed approach in practice. Originality/value – This paper draws attention to ALB problem in which workers have been assigned to teams in advance due to the need for specialized skills or equipment on the line for the first time.

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