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https://www.readbyqxmd.com/read/28927539/transient-and-stable-vector-transfection-pitfalls-off-target-effects-artifacts
#1
REVIEW
Aleksei A Stepanenko, Henry H Heng
Transient and stable vector transfections have played important roles in illustrating the function of specific genes/proteins. The general assumption is that such a platform could effectively link a given gene/protein to gained phenotypes, revealing the mechanism of how a gene works. However, in reality, increased studies have surprisingly noticed some unexpected results. In this review, we demonstrate that an assumption that empty vector-transfected cells preserve the cytogenetic and phenotypic characteristics, and represent the adequate control in transfection experiments is not universally valid...
July 2017: Mutation Research
https://www.readbyqxmd.com/read/28925669/lower-extremity-abnormalities-in-children
#2
Caitlyn M Rerucha, Caleb Dickison, Drew C Baird
Leg and foot problems in childhood are common causes of parental concern. Rotational problems include intoeing and out-toeing. Intoeing is most common in infants and young children. Intoeing is caused by metatarsus adductus, internal tibial torsion, and femoral anteversion. Out-toeing is less common than intoeing and occurs more often in older children. Out-toeing is caused by external tibial torsion and femoral retroversion. Angular problems include genu varum (bowleg) and genu valgum (knock knee). With pes planus (flatfoot), the arch of the foot is usually flexible rather than rigid...
August 15, 2017: American Family Physician
https://www.readbyqxmd.com/read/28915816/cramp-deficiency-leads-to-a-pro-inflammatory-phenotype-and-impaired-phagocytosis-after-exposure-to-bacterial-meningitis-pathogens
#3
Eugenia Kress, Julika Merres, Lea-Jessica Albrecht, Sven Hammerschmidt, Thomas Pufe, Simone C Tauber, Lars-Ove Brandenburg
BACKGROUND: Antimicrobial peptides are important components of the host defence with a broad range of functions including direct antimicrobial activity and modulation of inflammation. Lack of cathelin-related antimicrobial peptide (CRAMP) was associated with higher mortality and bacterial burden and impaired neutrophil granulocyte infiltration in a model of pneumococcal meningitis. The present study was designed to characterize the effects of CRAMP deficiency on glial response and phagocytosis after exposure to bacterial stimuli...
September 16, 2017: Cell Communication and Signaling: CCS
https://www.readbyqxmd.com/read/28912160/dopamine-d2-receptors-modulate-pyramidal-neurons-in-mouse-medial-prefrontal-cortex-through-a-stimulatory-g-protein-pathway
#4
Sarah E Robinson, Vikaas S Sohal
Dopaminergic modulation of prefrontal cortex is believed to play key roles in many cognitive functions and to be disrupted in pathological conditions such as schizophrenia. We have previously described a phenomenon whereby dopamine D2 receptor (D2R) activation elicits afterdepolarizations (ADPs) in subcortically-projecting (SC) pyramidal neurons within L5 of the prefrontal cortex. These D2R-induced ADPs only occur following synaptic input which activates NMDA receptors (NMDARs) even when the delay between the synaptic input and afterdepolarizations is relatively long, e...
September 14, 2017: Journal of Neuroscience: the Official Journal of the Society for Neuroscience
https://www.readbyqxmd.com/read/28901722/%C3%AE-6-subunit-containing-nicotinic-receptors-mediate-low-dose-ethanol-effects-on-ventral-tegmental-area-neurons-and-ethanol-reward
#5
Scott C Steffensen, Samuel I Shin, Ashley C Nelson, Stephanie S Pistorius, Stephanie B Williams, Taylor J Woodward, Hyun Jung Park, Lindsey Friend, Ming Gao, Fenfei Gao, Devin H Taylor, M Foster Olive, Jeffrey G Edwards, Sterling N Sudweeks, Lori M Buhlman, J Michael McIntosh, Jie Wu
Dopamine (DA) neuron excitability is regulated by inhibitory GABAergic synaptic transmission and modulated by nicotinic acetylcholine receptors (nAChRs). The aim of this study was to evaluate the role of α6 subunit-containing nAChRs (α6*-nAChRs) in acute ethanol effects on ventral tegmental area (VTA) GABA and DA neurons. α6*-nAChRs were visualized on GABA terminals on VTA GABA neurons, and α6*-nAChR transcripts were expressed in most DA neurons, but only a minority of VTA GABA neurons from GAD67 GFP mice...
September 13, 2017: Addiction Biology
https://www.readbyqxmd.com/read/28894266/generation-of-a-new-mouse-model-of-glaucoma-characterized-by-reduced-expression-of-the-ap-2%C3%AE-and-ap-2%C3%AE-proteins
#6
Maria Monica Barzago, Mami Kurosaki, Maddalena Fratelli, Marco Bolis, Chiara Giudice, Laura Nordio, Elisa Cerri, Luciano Domenici, Mineko Terao, Enrico Garattini
We generated 6 transgenic lines with insertion of an expression plasmid for the R883/M xanthine dehydrogenase (XDH) mutant protein. Approximately 20% of the animals deriving from one of the transgenic lines show ocular abnormalities and an increase in intra-ocular pressure which are consistent with glaucoma. The observed pathologic phenotype is not due to expression of the transgene, but rather the consequence of the transgene insertion site, which has been defined by genome sequencing. The insertion site maps to chromosome 1qA3 in close proximity to the loci encoding AP-2β and AP-2δ, two proteins expressed in the eye...
September 11, 2017: Scientific Reports
https://www.readbyqxmd.com/read/28893189/differentially-expressed-genes-during-the-imbibition-of-dormant-and-after-ripened-seeds-a-reverse-genetics-approach
#7
Farzaneh Yazdanpanah, Johannes Hanson, Henk W M Hilhorst, Leónie Bentsink
BACKGROUND: Seed dormancy, defined as the incapability of a viable seed to germinate under favourable conditions, is an important trait in nature and agriculture. Despite extensive research on dormancy and germination, many questions about the molecular mechanisms controlling these traits remain unanswered, likely due to its genetic complexity and the large environmental effects which are characteristic of these quantitative traits. To boost research towards revealing mechanisms in the control of seed dormancy and germination we depend on the identification of genes controlling those traits...
September 11, 2017: BMC Plant Biology
https://www.readbyqxmd.com/read/28887630/ncb5or-deficiency-in-the-cerebellum-and-midbrain-leads-to-dehydration-and-alterations-in-thirst-response-fasted-feeding-behavior-and-voluntary-exercise-in-mice
#8
Matthew A Stroh, Michelle K Winter, Kenneth E McCarson, John P Thyfault, Hao Zhu
Cytosolic NADH-cytochrome-b5-oxidoreductase (NCB5OR) is ubiquitously expressed in animal tissues. We have previously reported that global ablation of NCB5OR in mice results in early-onset lean diabetes with decreased serum leptin levels and increased metabolic and feeding activities. The conditional deletion of NCB5OR in the mouse cerebellum and midbrain (conditional knock out, CKO mice) results in local iron dyshomeostasis and altered locomotor activity. It has been established that lesion to or removal of the cerebellum leads to changes in nutrient organization, visceral response, feeding behavior, and body weight...
September 8, 2017: Cerebellum
https://www.readbyqxmd.com/read/28885142/sonic-hedgehog-switches-on-wnt-planar-cell-polarity-signaling-in-commissural-axon-growth-cones-by-reducing-levels-of-shisa2
#9
Keisuke Onishi, Yimin Zou
Commissural axons switch on responsiveness to Wnt attraction during midline crossing and turn anteriorly only after exiting the floor plate. We report here that Sonic Hedgehog (Shh)-Smoothened signaling downregulates Shisa2, which inhibits the glycosylation and cell surface presentation of Frizzled3 in rodent commissural axon growth cones. Constitutive Shisa2 expression causes randomized turning of post-crossing commissural axons along the anterior-posterior (A-P) axis. Loss of Shisa2 led to precocious anterior turning of commissural axons before or during midline crossing...
September 8, 2017: ELife
https://www.readbyqxmd.com/read/28883020/increased-gsnor-expression-during-aging-impairs-cognitive-function-and-decreases-s-nitrosation-of-camkii%C3%AE
#10
Yuying Zhang, Kaiyuan Wu, Wenting Su, Deng-Feng Zhang, Ping Wang, Xinhua Qiao, Qin Yao, Zengqiang Yuan, Yong-Gang Yao, Guanghui Liu, Chen Zhang, Limin Liu, Chen Chang
As the population ages, an increasing number of people suffer from age-related cognitive impairment. However, the mechanisms underlying this process remain unclear. Here, we found that S-nitrosoglutathione reductase (GSNOR), the key enzyme that metabolizes intracellular nitric oxide (NO) and regulates S-nitrosation, was significantly increased in the hippocampus of both aging humans and mice. Transgenic mice overexpressing GSNOR exclusively in neurons showed cognitive impairment in behavioral tests, including the Morris water maze, fear conditioning and the Y-maze test...
September 7, 2017: Journal of Neuroscience: the Official Journal of the Society for Neuroscience
https://www.readbyqxmd.com/read/28878098/loss-of-protein-arginine-methyltransferase-8-alters-synapse-composition-and-function-resulting-in-behavioral-defects
#11
Jay Penney, Jinsoo Seo, Oleg Kritskiy, Sara Elmsaouri, Fan Gao, Ping-Chieh Pao, Susan C Su, Li-Huei Tsai
Diverse molecular mechanisms regulate synaptic composition and function in the mammalian nervous system. The multifunctional protein arginine methyltransferase 8 (PRMT8) possesses both methyltransferase and phospholipase activities. Here we examine the role of this neuron-specific protein in hippocampal plasticity and cognitive function. PRMT8 protein localizes to synaptic sites, and conditional whole-brain Prmt8 deletion results in altered levels of multiple synaptic proteins in the hippocampus, using both male and female mice...
September 6, 2017: Journal of Neuroscience: the Official Journal of the Society for Neuroscience
https://www.readbyqxmd.com/read/28877240/runx1-mediates-the-development-of-the-granular-convoluted-tubules-in-the-submandibular-glands
#12
Hitomi Ono Minagi, Safiye Esra Sarper, Hiroshi Kurosaka, Koh-Ichi Kuremoto, Ichiro Taniuchi, Takayoshi Sakai, Takashi Yamashiro
The mouse granular convoluted tubules (GCTs), which are only located in the submandibular gland (SMG) are known to develop and maintain their structure in an androgen-dependent manner. We previously demonstrated that the GCTs are involuted by the epithelial deletion of core binding factor β (CBFβ), a transcription factor that physically interacts with any of the Runt-related transcription factor (RUNX) proteins (RUNX1, 2 and 3). This result clearly demonstrates that the Runx /Cbfb signaling pathway is indispensable in the development of the GCTs...
2017: PloS One
https://www.readbyqxmd.com/read/28874552/global-transcriptional-regulatory-network-for-escherichia-coli-robustly-connects-gene-expression-to-transcription-factor-activities
#13
Xin Fang, Anand Sastry, Nathan Mih, Donghyuk Kim, Justin Tan, James T Yurkovich, Colton J Lloyd, Ye Gao, Laurence Yang, Bernhard O Palsson
Transcriptional regulatory networks (TRNs) have been studied intensely for >25 y. Yet, even for the Escherichia coli TRN-probably the best characterized TRN-several questions remain. Here, we address three questions: (i) How complete is our knowledge of the E. coli TRN; (ii) how well can we predict gene expression using this TRN; and (iii) how robust is our understanding of the TRN? First, we reconstructed a high-confidence TRN (hiTRN) consisting of 147 transcription factors (TFs) regulating 1,538 transcription units (TUs) encoding 1,764 genes...
September 5, 2017: Proceedings of the National Academy of Sciences of the United States of America
https://www.readbyqxmd.com/read/28866324/effects-of-in-vivo-deletion-of-gata2-in-bone-marrow-stromal-cells
#14
Shin Hasegawa, Tohru Fujiwara, Yoko Okitsu, Hiroki Kato, Yuki Sato, Noriko Fukuhara, Yasushi Onishi, Ritsuko Shimizu, Masayuki Yamamoto, Hideo Harigae
The bone marrow (BM) microenvironment comprises multiple stem cell niches derived from BM mesenchymal stem cells (BM-MSCs). Previous in vitro analyses have suggested that transcription factor GATA2 plays an important role in adipocyte differentiation of BM-mesenchymal stem cells as well as in hematopoietic support, but the role of GATA2 in vivo remains unknown. We evaluated GATA2 effects in BM-MSC in vivo. Expression profiling analysis of Gata2 knockout Ter119(-)CD45(-) mesenchymal stromal cells obtained from compact bone from tamoxifen-treated Gata2 conditional knockout mice (Gata2(f/f)/ER-Cre mice) revealed upregulation of 110 genes and downregulation of 141 by a factor of 2...
August 30, 2017: Experimental Hematology
https://www.readbyqxmd.com/read/28863347/deletion-of-dlk2-increases-the-vulnerability-to-anxiety-like-behaviors-and-impairs-the-anxiolytic-action-of-alprazolam
#15
Francisco Navarrete, María S García-Gutiérrez, Jorge Laborda, Jorge Manzanares
The purpose of this study was to evaluate the role of the non-canonical DLK2 NOTCH ligand in the regulation of emotional behavior. To this aim, anxiety and depressive-like behaviors were examined in Dlk2 knock-out (Dlk2(-/-)) and its corresponding wild-type (WT) mice. Furthermore, relative gene expression analyses of corticotropin releasing hormone (Crh) in the paraventricular nucleus (PVN), glucocorticoid receptor (NR3C1) and FK506 binding protein 5 (FKBP5) in the hippocampus (HIPP), and the transcription factors Hes1, Hes5 and Hey1 in the PVN, HIPP and amygdala (AMY) were carried out in Dlk2(-/-) and WT mice under basal conditions and after exposure to restraint stress...
August 29, 2017: Psychoneuroendocrinology
https://www.readbyqxmd.com/read/28851850/toxoplasma-gondii-requires-glycogen-phosphorylase-for-balancing-amylopectin-storage-and-for-efficient-production-of-brain-cysts
#16
Tatsuki Sugi, Vincent Tu, Yanfen Ma, Tadakimi Tomita, Louis M Weiss
In immunocompromised hosts, latent infection with Toxoplasma gondii can reactivate from tissue cysts, leading to encephalitis. A characteristic of T. gondii bradyzoites in tissue cysts is the presence of amylopectin granules. The regulatory mechanisms and role of amylopectin accumulation in this organism are not fully understood. The T. gondii genome encodes a putative glycogen phosphorylase (TgGP), and mutants were constructed to manipulate the activity of TgGP and to evaluate the function of TgGP in amylopectin storage...
August 29, 2017: MBio
https://www.readbyqxmd.com/read/28842684/plasmodium-falciparum-gpcr-like-receptor-sr25-mediates-extracellular-k-sensing-coupled-to-ca-2-signaling-and-stress-survival
#17
Miriam S Moraes, Alexandre Budu, Maneesh K Singh, Lucas Borges-Pereira, Julio Levano-Garcia, Chiara Currà, Leonardo Picci, Tomasino Pace, Marta Ponzi, Tullio Pozzan, Célia R S Garcia
The malaria parasite Plasmodium falciparum is exposed, during its development, to major changes of ionic composition in its surrounding medium. We demonstrate that the P. falciparum serpentine-like receptor PfSR25 is a monovalent cation sensor capable of modulating Ca(2+) signaling in the parasites. Changing from high (140 mM) to low (5.4 mM) KCl concentration triggers [Ca(2+)]cyt increase in isolated parasites and this Ca(2+) rise is blocked either by phospholipase C (PLC) inhibition or by depleting the parasite's internal Ca(2+) pools...
August 25, 2017: Scientific Reports
https://www.readbyqxmd.com/read/28833131/the-core-planar-cell-polarity-gene-vangl2-maintains-apical-basal-organisation-of-the-corneal-epithelium
#18
D Alessio Panzica, Amy S Findlay, Rianne van Ladesteijn, J Martin Collinson
The role of the core planar cell polarity (PCP) pathway protein, Vangl2, was investigated in the corneal epithelium of the mammalian eye, a paradigm anatomical model of planar cell migration. The gene was conditionally knocked out in vivo and knocked down by siRNA, followed by immunohistochemical, behavioural and morphological analysis of corneal epithelial cells. The primary defects observed in vivo were of apical-basal organisation of the corneal epithelium, with abnormal stratification throughout life, mislocalisation of the cell membrane protein, Scribble, to the basal side of cells, and partial loss of the epithelial basement membrane...
August 17, 2017: Journal of Anatomy
https://www.readbyqxmd.com/read/28820880/knock-out-of-a-mitochondrial-sirtuin-protects-neurons-from-degeneration-in-caenorhabditis-elegans
#19
Rachele Sangaletti, Massimo D'Amico, Jeff Grant, David Della-Morte, Laura Bianchi
Sirtuins are NAD⁺-dependent deacetylases, lipoamidases, and ADP-ribosyltransferases that link cellular metabolism to multiple intracellular pathways that influence processes as diverse as cell survival, longevity, and cancer growth. Sirtuins influence the extent of neuronal death in stroke. However, different sirtuins appear to have opposite roles in neuronal protection. In Caenorhabditis elegans, we found that knock-out of mitochondrial sirtuin sir-2.3, homologous to mammalian SIRT4, is protective in both chemical ischemia and hyperactive channel induced necrosis...
August 2017: PLoS Genetics
https://www.readbyqxmd.com/read/28818866/systematic-identification-of-determinants-for-single-strand-annealing-mediated-deletion-formation-in-saccharomyces-cerevisiae
#20
Maia Segura-Wang, Megumi Onishi-Seebacher, Adrian M Stütz, Balca R Mardin, Jan O Korbel
To ensure genomic integrity, living organisms have evolved diverse molecular processes for sensing and repairing damaged DNA. If improperly repaired, DNA damage can give rise to different types of mutations, an important class of which are the genomic structural variants (SVs). In spite of their importance for phenotypic variation and genome evolution, potential contributors to SV formation in Saccharomyces cerevisiae (budding yeast), a highly tractable model organism, are not fully recognized. Here we developed and applied a genome-wide assay to identify yeast gene knock-out mutants associated with de novo deletion formation, in particular single strand annealing (SSA) -mediated deletion formation in a systematic manner...
August 17, 2017: G3: Genes—Genomes—Genetics
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