Early transcription from the maternal genome controlling blastomere integrity in mouse two-cell-stage embryos

Blastomere cytofragmentation in mammalian embryos poses a significant problem in applied and clinical embryology. Mouse two-cell-stage embryos display strain-dependent differences in the rate of cytofragmentation, with a high rate observed in C3H/HeJ embryos and a lower rate observed in C57BL/6 embryos. The maternally inherited genome exerts the strongest effect on the process, with lesser effects mediated by the paternally inherited genome and the ooplasm. The effect of the maternal genome is transcription dependent and independent of the mitochondrial strain of origin. To identify molecular mechanisms that underlie cytofragmentation, we evaluated transcriptional activities of embryos possessing maternal pronuclei (mPN) of different origins. The mPN from C57BL/6 and C3H/HeJ strains directed specific transcription at the two-cell stage of mRNAs corresponding to 935 and 864 Affymetrix probe set IDs, respectively. Comparing transcriptomes of two-cell-stage embryos with different mPN revealed 64 transcribed genes with differential expression (1.4-fold or greater). Some of these genes occupy molecular pathways that may regulate cytofragmentation via a combination of effects related to apoptosis and effects on the cytoskeleton. These results implicate specific molecular mechanisms that may regulate cytofragmentation in early mammalian embryos. The most striking effect of mPN strain of origin on gene expression was on adenylate cyclase 2 ( Adcy2). Treatment with dibutyryl cAMP (dbcAMP) elicits a high rate and severe form of cytofragmentation, and the effective dbcAMP concentration varies with maternal genotype. An activator of exchange proteins directly activated by cAMP (EPACs, or RAPGEF 3 and 4) 8-pCPT-2′- O-methyl-cAMP, elicits a high level of fragmentation while the PKA-specific activator N6-benzoyl-cAMP does not. Inhibition of A kinase anchor protein activities with st-Ht31 induces fragmentation. Inhibition of phosphatidylinositol 3-kinase signaling also induces fragmentation. These results reveal novel mechanisms by which maternal genotype affects cytofragmentation, including a system of opposing signaling pathways that most likely operate by controlling cytoskeletal function.

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Transcription factor/DNA interactions visualized by electron spectroscopic imaging

Proceedings, annual meeting, Electron Microscopy Society of America ◽

10.1017/s0424820100122654 ◽

1992 ◽

Vol 50 (1) ◽

pp. 450-451

Author(s):

David P. Bazett-Jones ◽

Mark L. Brown

Keyword(s):

Transcription Factor ◽

Dna Sequences ◽

Transcription Initiation ◽

Molecular Mechanisms ◽

Phosphorus Content ◽

Spectroscopic Imaging ◽

Electron Spectroscopic Imaging ◽

Dna Backbone ◽

Two Parameters ◽

High Level

A multisubunit RNA polymerase enzyme is ultimately responsible for transcription initiation and elongation of RNA, but recognition of the proper start site by the enzyme is regulated by general, temporal and gene-specific trans-factors interacting at promoter and enhancer DNA sequences. To understand the molecular mechanisms which precisely regulate the transcription initiation event, it is crucial to elucidate the structure of the transcription factor/DNA complexes involved. Electron spectroscopic imaging (ESI) provides the opportunity to visualize individual DNA molecules. Enhancement of DNA contrast with ESI is accomplished by imaging with electrons that have interacted with inner shell electrons of phosphorus in the DNA backbone. Phosphorus detection at this intermediately high level of resolution (≈lnm) permits selective imaging of the DNA, to determine whether the protein factors compact, bend or wrap the DNA. Simultaneously, mass analysis and phosphorus content can be measured quantitatively, using adjacent DNA or tobacco mosaic virus (TMV) as mass and phosphorus standards. These two parameters provide stoichiometric information relating the ratios of protein:DNA content.

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par-4, a gene required for cytoplasmic localization and determination of specific cell types in Caenorhabditis elegans embryogenesis.

Genetics ◽

10.1093/genetics/130.4.771 ◽

1992 ◽

Vol 130 (4) ◽

pp. 771-790 ◽

Cited By ~ 5

Author(s):

D G Morton ◽

J M Roos ◽

K J Kemphues

Keyword(s):

Caenorhabditis Elegans ◽

Cell Types ◽

Intestinal Cells ◽

Specific Cell ◽

Cytoplasmic Localization ◽

Loss Of Function ◽

Embryo Viability ◽

Cell Stage ◽

Maternal Genome

Abstract Specification of some cell fates in the early Caenorhabditis elegans embryo is mediated by cytoplasmic localization under control of the maternal genome. Using nine newly isolated mutations, and two existing mutations, we have analyzed the role of the maternally expressed gene par-4 in cytoplasmic localization. We recovered seven new par-4 alleles in screens for maternal effect lethal mutations that result in failure to differentiate intestinal cells. Two additional par-4 mutations were identified in noncomplementation screens using strains with a high frequency of transposon mobility. All 11 mutations cause defects early in development of embryos produced by homozygous mutant mothers. Analysis with a deficiency in the region indicates that it33 is a strong loss-of-function mutation. par-4(it33) terminal stage embryos contain many cells, but show no morphogenesis, and are lacking intestinal cells. Temperature shifts with the it57ts allele suggest that the critical period for both intestinal differentiation and embryo viability begins during oogenesis, about 1.5 hr before fertilization, and ends before the four-cell stage. We propose that the primary function of the par-4 gene is to act as part of a maternally encoded system for cytoplasmic localization in the first cell cycle, with par-4 playing a particularly important role in the determination of intestine. Analysis of a par-4; par-2 double mutant suggests that par-4 and par-2 gene products interact in this system.

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Investigating the resistance levels and mechanisms to penoxsulam and cyhalofop-butyl in barnyardgrass (Echinochloa crus-galli) from Ningxia province, China

Weed Science ◽

10.1017/wsc.2021.37 ◽

2021 ◽

pp. 1-25

Author(s):

Qian Yang ◽

Xia Yang ◽

Zichang Zhang ◽

Jieping Wang ◽

Weiguo Fu ◽

...

Keyword(s):

Herbicide Resistance ◽

Molecular Mechanisms ◽

Acetolactate Synthase ◽

Rice Fields ◽

Target Site ◽

Herbicide Resistant ◽

Susceptible Populations ◽

High Level ◽

Als Inhibitor ◽

Encoding Genes

Abstract Barnyardgrass (Echinochloa crus-galli) is a noxious grass weed which infests rice fields and causes huge crop yield losses. In this study, we collected twelve E. crus-galli populations from rice ﬁelds of Ningxia province in China and investigated the resistance levels to acetolactate synthase (ALS) inhibitor penoxsulam and acetyl-CoA carboxylase (ACCase) inhibitor cyhalofop-butyl. The results showed that eight populations exhibited resistance to penoxsulam and four populations evolved resistance to cyhalofop-butyl. Moreover, all of the four cyhalofop-butyl-resistant populations (NX3, NX4, NX6 and NX7) displayed multiple-herbicide-resistance (MHR) to both penoxsulam and cyhalofop-butyl. The alternative herbicides bispyribac-sodium, metamifop and fenoxaprop-P-ethyl cannot effectively control the MHR plants. To characterize the molecular mechanisms of resistance, we ampliﬁed and sequenced the target-site encoding genes in resistant and susceptible populations. Partial sequences of three ALS genes and six ACCase genes were examined. A Trp-574-Leu mutation was detected in EcALS1 and EcALS3 in two high-level (65.84- and 59.30-fold) penoxsulam-resistant populations NX2 and NX10, respectively. In addition, one copy (EcACC4) of ACCase genes encodes a truncated aberrant protein due to a frameshift mutation in E. crus-galli populations. None of amino acid substitutions that are known to confer herbicide resistance were detected in ALS and ACCase genes of MHR populations. Our study reveals the widespread of multiple-herbicide resistant E. crus-galli populations at Ningxia province of China that exhibit resistance to several ALS and ACCase inhibitors. Non-target-site based mechanisms are likely to be involved in E. crus-galli resistance to the herbicides, at least in four MHR populations.

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High-Intensity Focused Ultrasound Circular Cyclocoagulation in Glaucoma: A Step Forward for Cyclodestruction?

Journal of Ophthalmology ◽

10.1155/2017/7136275 ◽

2017 ◽

Vol 2017 ◽

pp. 1-14 ◽

Cited By ~ 5

Author(s):

Rodolfo Mastropasqua ◽

Vincenzo Fasanella ◽

Alessandra Mastropasqua ◽

Marco Ciancaglini ◽

Luca Agnifili

Keyword(s):

High Intensity ◽

Ciliary Body ◽

Focused Ultrasound ◽

Current Knowledge ◽

High Intensity Focused Ultrasound ◽

High Rate ◽

Dose Effect ◽

Computer Assisted ◽

Refractory Glaucoma ◽

High Level

The ciliary body ablation is still considered as a last resort treatment to reduce the intraocular pressure (IOP) in uncontrolled glaucoma. Several ablation techniques have been proposed over the years, all presenting a high rate of complications, nonselectivity for the target organ, and unpredictable dose-effect relationship. These drawbacks limited the application of cyclodestructive procedures almost exclusively to refractory glaucoma. High-intensity focused ultrasound (HIFU), proposed in the early 1980s and later abandoned because of the complexity and side effects of the procedure, was recently reconsidered in a new approach to destroy the ciliary body. Ultrasound circular cyclocoagulation (UC3), by using miniaturized transducers embedded in a dedicated circular-shaped device, permits to selectively treat the ciliary body in a one-step, computer-assisted, and non-operator-dependent procedure. UC3 shows a high level of safety along with a predictable and sustained IOP reduction in patients with refractory glaucoma. Because of this, the indication of UC3 was recently extended also to naïve-to-surgery patients, thus reconsidering the role and timing of ciliary body ablation in the surgical management of glaucoma. This article provides a review of the most used cycloablative techniques with particular attention to UC3, summarizing the current knowledge about this procedure and future possible developments.

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708 A NEW DIAGNOSTIC PARADIGM FOR LARYNGOPHARYNGEAL REFLUX DISEASE: CORRELATION OF IMPEDANCE-PH MONITORING AND DIGITAL REFLUX SCINTIGRAPHY RESULTS

Diseases of the Esophagus ◽

10.1093/dote/doab052.708 ◽

2021 ◽

Vol 34 (Supplement_1) ◽

Author(s):

Jin-soo Park ◽

Oleksandr Khoma ◽

Hans Van Der Wall ◽

Gregory Falk

Keyword(s):

Laryngopharyngeal Reflux ◽

Ph Monitoring ◽

Acid Reflux ◽

High Rate ◽

High Yield ◽

Time Activity ◽

Proximal Esophagus ◽

Laryngopharyngeal Reflux Disease ◽

High Level ◽

No Gold Standard

Abstract No gold-standard investigation exists for laryngopharyngeal reflux (LPR). Multichannel intraluminal impedance (MII)-pH testing has uncertain utility in LPR. Meanwhile, reflux scintigraphy allows immediate and delayed visualisation of tracer reflux in the esophagus, pharynx, and lungs. The present study aimed to correlate MII-pH and scintigraphic reflux results in patients with primary LPR. Methods Consecutive patients with LPR underwent MII-pH and scintigraphic reflux studies. Abnormal values for MII-pH results were defined from existing literature. MII-pH and scintigraphic data were correlated. Results 105 patients with LPR (31 males (29.5%), median age 60 years (range: 20–87)) were studied. Scintigraphic reflux was seen in the pharynx in 94 (90.4%), and in the proximal esophagus in 94 (90.4%). Delayed scintigraphic contamination of the pharynx was seen in 101 patients (96.2%) and in the lungs of 56 patients (53.3%). Abnormal reflux was seen in the distal esophagus in 12.4%, proximal esophagus in 25.7%, and in the pharynx in 82.9%. Patients with poor scintigraphic clearance had higher Demeester scores (p = 0.043), more proximal reflux episodes (p = 0.046), more distal acid reflux episodes (p = 0.023), and longer bolus clearance times (p = 0.002). Conclusion Reflux scintigraphy has a high yield in LPR patients. Scintigraphic time-activity curves correlated with validated MII-pH results. A high rate of pulmonary microaspiration was found in LPR patients. This study demonstrated a high level of pharyngeal contamination by scintigraphy and MII-pH, which supports the use of digital reflux scintigraphy in diagnosing LPR.

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Comparative assessment of the dynamics of the incidence of pulmonary and etraulmonary tuberculosis in St. Petersburg for half a century of observation

MedAlliance ◽

10.36422//23076348-2020-8-3-6-14 ◽

2020 ◽

Vol 8 (3) ◽

pp. 6-14

Keyword(s):

Control System ◽

Incidence Rate ◽

Extrapulmonary Tuberculosis ◽

High Rate ◽

Tb Control ◽

Complete Restoration ◽

Third Stage ◽

High Level ◽

Clinical Awareness ◽

Positive Effect

This article describes the results of comparative analysis of the dynamics of incidence of pulmonary tuberculo-sis (TBP), extrapulmonary extrarespiratory tuberculo-sis (TBER) and respiratory extrapulmonary tuberculosis (TBREP) in St. Petersburg (Leningrad) from 1970 to 2019. Throughout the 50 years of observation, epidemiological situation for TBP, especially for TBER, hasmuch improved, with incidence of the latter going down 30 times. Five stages (decades) were identified, within which the factors demonstrate practically the same impact, the growth rate of indicators was calculated for each stage (the value of indicators at the beginning of each stage taken as 100%).The first stage was a simultaneous reduction in the inci-dence rate in all categories of TB localization against the background of socio-economic factors positive effect and organization of effective and comprehensive an-ti-TB measures. In the second stage (1980ies, a period of growing socio-economic problems), the positive dynam-ics of TB incidence stopped and switched to stabilization at a low level. Indicators became more volatile, while TB dynamics by localization more and more desynchro-nized. Тhe third stage (1990iеs, a period of social and economic crisis) was char-acterised by a sharp increase of TB incidence, especially TBREP, with a switch to stabi-lization at a high level. The fourth stage (2000 through 2009, the beginning of TB control system restoration) demonstrated the indicators’ dynamics desynchroniza-tion: the incidence of TBP and TBREP has stabilized, and TBER continued to decline. The fifth stage (2009 through 2019, complete restoration of TB control system) showed a steady trend of incidence rate reduction and return of uniformity in incidence rate of various localizations.There is no increase in the proportion of extrapulmonary TB expected for the incidence decline. TBER incidence had been declining in most stages, with the exception of the 90ies, which could be explained by substandard work in identifying patients, and the rarity of TBER etiological verification. It is necessary to raise the clinical awareness of narrow specialists and general practitioners in the di-agnostic problems associated with TBER.The coincidence of the rate of TB incidence decrease in main localizations in the 1970ies and 2010s proves that a comprehensive state strategy to combat tuberculosis can provide a high rate of TB incidence decrease in various social layers with positive social and economic conditions of life of the population.

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Truncated Non-Nuclear Transposable Elements in Grapevine: A Mini Review

Scientia Agriculturae Bohemica ◽

10.2478/sab-2019-0030 ◽

2019 ◽

Vol 50 (4) ◽

pp. 219-227

Author(s):

A.V. Milovanov ◽

J. Tello ◽

U.C.M. Anhalt ◽

A. Forneck

Keyword(s):

Gene Transfer ◽

Mitochondrial Genome ◽

Transposable Elements ◽

Vitis Vinifera ◽

In Silico ◽

Gene Evolution ◽

High Rate ◽

Miniature Inverted Transposable Elements ◽

High Level ◽

Inner Region

Abstract In this mini-review we present insight to the non-nuclear transposable elements and in silico analysis of miniature inverted transposable elements (MITEs) in the grapevine mitochondrial genome. Here we report the identification of 17 truncated sequences in grapevine (Vitis vinifera L.) mitochondrial genome which expectedly belongs to the four ancient transposon families (hAT, Tc1Mariner, Mutator and PIF/Harbinger). Some sequences with a high rate of homology in chloroplast and nuclear genomes were also identified. Thus, it suggests the intercellular gene transfer between these three organelles. These partial sequences showed a high level of similitude with full MITE sequences, and they were found in their inner region, supporting their MITE origin. Further analysis revealed these sequences in other life kingdoms (including eubacteria and archaea), which indicates their ancient origin. Further research showed that 13 out of the 17 sequences are conserved domains of the genes where they are located, suggesting their contribution to gene evolution. Therefore, we suppose that more studies of nature, origin and functional meaning of these sequences and their fusion with genes are necessary. In the light of our observations it will be useful for further studies of V. vinifera genome organizing and systematics, as well as for other species.

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Proteomic Differences in the Hippocampus and Cortex of Epilepsy Brain Tissue

10.1101/2020.07.21.209163 ◽

2020 ◽

Cited By ~ 1

Author(s):

Geoffrey Pires ◽

Dominique Leitner ◽

Eleanor Drummond ◽

Evgeny Kanshin ◽

Shruti Nayak ◽

...

Keyword(s):

Dentate Gyrus ◽

Frontal Cortex ◽

G Protein ◽

Molecular Mechanisms ◽

High Rate ◽

Label Free ◽

Protein Subunit ◽

Unexpected Death ◽

Hippocampal Ca1 ◽

And Control

AbstractEpilepsy is a common neurological disorder affecting over 70 million people worldwide, with a high rate of pharmaco-resistance, diverse comorbidities including progressive cognitive and behavioral disorders, and increased mortality from direct (e.g., Sudden Unexpected Death in Epilepsy [SUDEP], accidents, drowning) or indirect effects of seizures and therapies. Extensive research with animal models and human studies provides limited insights into the mechanisms underlying seizures and epileptogenesis, and these have not translated into significant reductions in pharmaco-resistance, morbidities or mortality. To help define changes in molecular signaling networks associated with epilepsy, we examined the proteome of brain samples from epilepsy and control cases. Label-free quantitative mass spectrometry (MS) was performed on the hippocampal CA1-3 region, frontal cortex, and dentate gyrus microdissected from epilepsy and control cases (n=14/group). Epilepsy cases had significant differences in the expression of 777 proteins in the hippocampal CA1-3 region, 296 proteins in the frontal cortex, and 49 proteins in the dentate gyrus in comparison to control cases. Network analysis showed that proteins involved in protein synthesis, mitochondrial function, G-protein signaling, and synaptic plasticity were particularly altered in epilepsy. While protein differences were most pronounced in the hippocampus, similar changes were observed in other brain regions indicating broad proteomic abnormalities in epilepsy. Among the most significantly altered proteins, G-protein Subunit Beta 1 (GNB1) was one of the most significantly decreased proteins in epilepsy in all regions studied, highlighting the importance of G-protein subunit signaling and G-protein–coupled receptors (GPCRs) in epilepsy. Our results provide insights into the molecular mechanisms underlying epilepsy, which may allow for novel targeted therapeutic strategies.

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The Urgency of Political Commitment in Formulation Reproductive Health Education Policy

Journal of Educational and Social Research ◽

10.36941/jesr-2021-0125 ◽

2021 ◽

Vol 11 (6) ◽

pp. 22

Author(s):

Margareth Christine ◽

Caroline Paskarina

Keyword(s):

Health Education ◽

Reproductive Health ◽

Maternal Mortality Ratio ◽

Public Health Education ◽

High Rate ◽

Political Commitment ◽

Reproductive Health Education ◽

Mortality Ratio ◽

High Level ◽

Special Region

Kulon Progo is the first area to have a local content of Reproductive Health Education in the curriculum, that is, since 2014. This was motivated by the high rate of underage marriage and the high level of maternal mortality ratio in the Special Region of Yogyakarta at that time. The Regent of Kulon Progo who served during that period, Hasto Wardoyo, was the party who initiated and directly led this policy-making process. This study seeks to analyze the form of political commitment from Hasto Wardoyo in the policy on Public Health Education by using Boli’s theory of political will criteria, in which political commitment is broadly divided into three groups, namely verbal will, institutional will, and budgetary will. Researchers used qualitative research methods supported by interview data with parties directly involved in the process of making and implementing policies, including the Chair of the Indonesia Union of Teachers, the Chair of the Indonesia Association of Family Planning, the Head of the Education Office, and the Head of the Health Office in the regency. The results of this study indicate that the Regent has shown his political commitment in the sense of verbal and regulation dimensions. However, in terms of institutional and budgetary will, the Regent did not show any form of political commitment, which resulted in the neglect of this policy on Reproductive Health Education after it had been launched. Received: 20 May 2021 / Accepted: 10 July 2021 / Published: 5 November 2021

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Phosphatidylinositol 3-kinase regulatory subunit 1 and phosphatase and tensin homolog as therapeutic targets in breast cancer

Tumor Biology ◽

10.1177/1010428317695529 ◽

2017 ◽

Vol 39 (3) ◽

pp. 101042831769552 ◽

Cited By ~ 1

Author(s):

Ebubekir Dirican ◽

Mustafa Akkiprik

Keyword(s):

Breast Cancer ◽

Cancer Progression ◽

Molecular Mechanisms ◽

Breast Cancer Progression ◽

Regulatory Subunit ◽

Phosphatidylinositol 3 Kinase ◽

Phosphatase And Tensin Homolog ◽

Tensin Homolog ◽

Kinase Pathway ◽

Phosphatidylinositol 3

Breast cancer is the most commonly diagnosed cancer among women in Turkey and worldwide. It is considered a heterogeneous disease and has different subtypes. Moreover, breast cancer has different molecular characteristics, behaviors, and responses to treatment. Advances in the understanding of the molecular mechanisms implicated in breast cancer progression have led to the identification of many potential therapeutic gene targets, such as Breast Cancer 1/2, phosphatidylinositol 3-kinase catalytic subunit alpha, and tumor protein 53. The aim of this review is to summarize the roles of phosphatidylinositol 3-kinase regulatory subunit 1 (alpha) (alias p85α) and phosphatase and tensin homolog in breast cancer progression and the molecular mechanisms involved. Phosphatase and tensin homolog is a tumor suppressor gene and protein. Phosphatase and tensin homolog antagonizes the phosphatidylinositol 3-kinase/AKT signaling pathway that plays a key role in cell growth, differentiation, and survival. Loss of phosphatase and tensin homolog expression, detected in about 20%–30% of cases, is known to be one of the most common tumor changes leading to phosphatidylinositol 3-kinase pathway activation in breast cancer. Instead, the regulatory subunit p85α is a significant component of the phosphatidylinositol 3-kinase pathway, and it has been proposed that a reduction in p85α protein would lead to decreased negative regulation of phosphatidylinositol 3-kinase and hyperactivation of the phosphatidylinositol 3-kinase pathway. Phosphatidylinositol 3-kinase regulatory subunit 1 protein has also been reported to be a positive regulator of phosphatase and tensin homolog via the stabilization of this protein. A functional genetic alteration of phosphatidylinositol 3-kinase regulatory subunit 1 that results in reduced p85α protein expression and increased insulin receptor substrate 1 binding would lead to enhanced phosphatidylinositol 3-kinase signaling and hence cancer development. Phosphatidylinositol 3-kinase regulatory subunit 1 underexpression was observed in 61.8% of breast cancer samples. Therefore, expression/alternations of phosphatidylinositol 3-kinase regulatory subunit 1 and phosphatase and tensin homolog genes have crucial roles for breast cancer progression. This review will summarize the biological roles of phosphatidylinositol 3-kinase regulatory subunit 1 and phosphatase and tensin homolog in breast cancer, with an emphasis on recent findings and the potential of phosphatidylinositol 3-kinase regulatory subunit 1 and phosphatase and tensin homolog as a therapeutic target for breast cancer therapy.

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