Here, we report that CRISPR guide RNAs (gRNAs) with a 5′-triphosphate group (5′-ppp gRNAs) produced via in vitro transcription trigger RNA-sensing innate immune responses human and murine cells, leading to cytotoxicity. 5′-ppp gRNAs the cytosol are recognized by DDX58, which turn activates type I interferon responses, causing up ∼80% cell death. We show triphosphate can be removed phosphatase resulting 5′-hydroxyl complex Cas9 or Cpf1 avoid achieve targeted mutagenesis at frequency of 95%...

OBJECT Chloroquine (CQ) is a quinoline-based drug widely used for the prevention and treatment of malaria. More recent studies have provided evidence that this may also harbor antitumor properties, whereby CQ possesses ability to accumulate in lysosomes blocks cellular process autophagy. Therefore, authors study set out investigate whether analogs, particular clinically established antimalaria drugs, would be able exert with specific focus on glioma cells. METHODS Toward goal, treated...

In a recent clinical trial, patients with newly diagnosed glioblastoma multiforme benefited from chloroquine (CQ) in combination conventional therapy (resection, temozolomide [TMZ], and radiation therapy). the present study, authors report mechanism by which CQ enhances therapeutic efficacy of TMZ to aid future studies aimed at improving this regimen.Using vitro vivo experiments, determined cytotoxicity. They focused on inhibition-of-autophagy knockdown autophagy-associated proteins or...

Abstract Formation of reactive oxygen and nitrogen species is a precipitating event in an array neuropathological conditions. In response to excessive (ROS) levels, transcriptionally dependent mechanisms drive the up‐regulation ROS scavenging proteins which, turn, limit extent brain damage. Here, we employed transgenic approach which cAMP‐response element binding protein (CREB)‐mediated transcription repressed (via A‐CREB) examine contribution CREB/cAMP pathway neuroprotection its potential...

Adult progenitor cell proliferation in the subgranular zone (SGZ) of dentate gyrus is a dynamic process that modulated by an array physiological process, including locomotor activity and novel environmental stimuli. In addition, pathophysiological events, such as ischemia status epilepticus (SE), have been shown to stimulate neurogenesis. Currently, limited information available regarding extracellular stimuli, receptors, downstream intracellular effectors couple excitotoxic stimulation...

Approximately 15% of non-small cell lung cancer cases are associated with a mutation in the epidermal growth factor receptor (EGFR) gene, which plays critical role tumor progression. With goal treating mutated EGFR-mediated cancer, we demonstrate use clustered regularly interspaced short palindromic repeats (CRISPR)/CRISPR protein 9 (Cas9) system to discriminate between oncogenic mutant and wild-type EGFR alleles eliminate carcinogenic allele high accuracy. We targeted an oncogene harboring...

Perillyl alcohol (POH) is a monoterpene that has been used orally for the treatment of systemic cancer. However, when significant gastrointestinal side effects and lack overall efficacy were documented. Recently, in phase II trial Brazil temozolomide (TMZ)-resistant malignant gliomas, POH was well tolerated administered intranasally. The present study explores mechanisms on TMZ-sensitive TMZ-resistant glioma cells. In vitro studies showed cytotoxic to as cells, this effect independent...

Duchenne muscular dystrophy (DMD) is a fatal, X-linked muscle-wasting disease caused by mutations in the DMD gene. In 51% of cases, reading frame disrupted because deletion several exons. Here, we show that CjCas9 derived from Campylobacter jejuni can be used as gene-editing tool to correct an out-of-frame Dmd exon knockout mice. Herein, Cas9 S. pyogenes generate mice with frameshift mutation Then, expressed CjCas9, its single-guide RNA, and EGFP gene tibialis anterior muscle using...

Glioblastoma (GBM) is the most aggressive primary brain tumor, exhibiting a high rate of recurrence and poor prognosis. Surgery chemoradiation with temozolomide (TMZ) represent standard care, but, in cases, tumor develops resistance to further treatment patients succumb disease. Therefore, there great need for development well-tolerated, effective drugs that specifically target chemoresistant gliomas. NEO214 was generated by covalently conjugating rolipram, PDE4 (phosphodiesterase 4)...

ABSTRACT Clostridium cellulovorans , an anaerobic bacterium, degrades native substrates efficiently by producing extracellular enzyme complex called the cellulosome. All cellulosomal subunits contain dockerin domains that can bind to hydrophobic termed cohesins which are repeated nine times in CbpA, nonenzymatic scaffolding protein of C. cellulosomes. In this study, synergistic interactions cellulases (endoglucanase E, EngE; endoglucanase L, EngL) and hemicellulases (arabinofuranosidase A,...

Excitotoxic cell death is one of the precipitating events in development temporal lobe epilepsy. Of particular prominence loss GABAergic hilar neurons. Although molecular mechanisms responsible for selective vulnerability these cells are not well understood, activation extracellular signal-regulated kinase/mitogen-activated protein kinase (ERK/MAPK) pathway has been implicated neuroprotective responses to excitotoxicity other neuronal populations. Here, we report that high levels...

Patients with glioblastoma multiforme (GBM), a malignant primary brain tumor, inevitably develop resistance to standard-of-care chemotherapy, temozolomide. This study explores the effects of novel agent NEO212, conjugate temozolomide perillyl alcohol, on temozolomide-resistant gliomas. NEO212 was tested for cytotoxic activity three human glioma cell lines, which were resistant based overexpression base excision repair (BER) pathway, mismatch (MMR) deficiency, or O(6) methyl-guanine-DNA...

The ability of two strains bacteria to cooperate in the synthesis an enzyme complex (a minicellulosome) was examined. Three Bacillus subtilis were constructed express Clostridium cellulovorans genes engB, xynB, and minicbpA. MiniCbpA, EngB, XynB synthesized secreted into medium by B. subtilis. When with minicbpA engB or xynB cocultured, minicellulosomes synthesized, consisting one case miniCbpA EngB second XynB. Both showed their respective enzymatic activities. We call this phenomenon...

There is no effective therapy for breast cancer that has spread to the brain. A major roadblock blood-brain barrier (BBB), which prevents usual drugs from effectively reaching intracranial metastases. The alkylating agent temozolomide (TMZ) able penetrate BBB and become gold standard chemotherapeutic treatment of glioblastoma. However, when it was tested in clinical trials activity against brain metastases cancer, results were mixed ranged "encouraging activity" "no objective responses." In...

ABSTRACT Two genes encoding EngB endoglucanase and mini-CbpA1 scaffolding protein of Clostridium cellulovorans were constructed coexpressed in Bacillus subtilis WB800. The resulting minicellulosomes isolated by gel filtration chromatography characterized. Biochemical immunological evidence indicated that fraction II contained consisting EngB. vivo synthesis suggests it will be possible the future to insert into B. cellulosomal allow growth on cellulosic materials production various designer...

Abstract The development of epilepsy is often associated with marked changes in central nervous system cell structure and function. Along these lines, reactive gliosis granule axonal sprouting within the dentate gyrus hippocampus are commonly observed individuals temporal lobe (TLE). Here we used pilocarpine model TLE mice to screen proteome phosphoproteome identify molecular events that altered as part pathogenic process. Using a two‐dimensional gel electrophoresis‐based approach, followed...

OBJECTIVE Glioblastoma (GBM) is the most prevalent and aggressive of primary brain tumors. There currently no effective treatment for this tumor. The proteasome inhibitor bortezomib a variety tumors, but not GBM. authors' goal was to demonstrate that can be in orthotopic GBM murine model if appropriate method drug delivery used. In study Alzet mini-osmotic pump used bring directly tumor brain, circumventing blood-brain barrier; thus making an METHODS 2 human glioma cell lines, U87 U251, were...

Abstract Background The antitumor efficacy of human epidermal growth factor receptor 2 (HER2)-targeted therapies, such as humanized monoclonal antibody trastuzumab (Herceptin®, Roche), in patients with breast-to-brain cancer metastasis is hindered by the low permeability blood-brain barrier (BBB). NEO100 a high-purity version natural monoterpene perillyl alcohol, produced under current good manufacturing practice (cGMP) regulations, that was shown previously to reversibly open BBB rodent...

(2000). The structure of the South Korean developmental regime and its transformation – statist mobilization authoritarian integration in anticommunist regimentation. Inter-Asia Cultural Studies: Vol. 1, No. 3, pp. 408-426.

Abstract Background Intracarotid injection of mannitol has been applied for decades to support brain entry therapeutics that otherwise do not effectively cross the blood–brain barrier (BBB). However, elaborate and high-risk nature this procedure kept its use restricted well-equipped medical centers. We are developing a more straightforward approach safely open BBB, based on intra-arterial (IA) NEO100, highly purified version natural monoterpene perillyl alcohol. Methods In vitro permeability...

The relation between the quaternary structure and substrate specificity of Thermus maltogenic amylase (ThMA) has been investigated. Sedimentation diffusion equilibrium ultracentrifugation gel filtration analyses, in combination with crystal determined recently, have demonstrated that ThMA existed a monomer/dimer equilibrium. truncation by removing N-terminal domain, which is composed 124 amino acid residues, resulted complete monomerization enzyme (ThMAΔ124) accompanied drastic decrease...