ABSTRACT
Recent advances in genome editing, especially CRISPR-Cas nucleases, have revolutionized both laboratory research and clinical therapeutics. CRISPR-Cas nucleases, together with the DNA damage repair pathway in cells, enable both genetic diversification by classical non-homologous end joining (c-NHEJ) and precise genome modification by homology-based repair (HBR). Genome editing in zygotes is a convenient way to edit the germline, paving the way for animal disease model generation, as well as human embryo genome editing therapy for some life-threatening and incurable diseases. HBR efficiency is highly dependent on the DNA donor that is utilized as a repair template. Here, we review recent progress in improving CRISPR-Cas nuclease-induced HBR in mammalian embryos by designing a suitable DNA donor. Moreover, we want to provide a guide for producing animal disease models and correcting genetic mutations through CRISPR-Cas nuclease-induced HBR in mammalian embryos. Finally, we discuss recent developments in precise genome-modification technology based on the CRISPR-Cas system.
Subject(s)
Animals , CRISPR-Cas Systems/genetics , DNA/genetics , Embryo, Mammalian/metabolism , Endonucleases/metabolism , Gene Editing , Mammals/metabolismABSTRACT
Citalopram is an antidepressant used for treating major depressive disorder. In the current work Citalopram HBr is formulated as mouth dissolving film with enhanced drug dissolution. The Central Composite Design (CCD), employed to examine the effects of amount of HPMC E50 (A), amount of maltodextrin (B) and amount of glycerol (C) on response variables tensile strength, disintegration time and cumulative % drug release. 27 formulations prepared according to CCD and evaluated for physicochemical parameters and in vitro dissolution studies. Citalopram HBr mouth dissolving films formulated by employing solvent-casting method using HPMC E50, maltodextrin and glycerol, optimized for the effective dosage of superdisintegrants. The formulation CF21 with maximum tensile strength of 67.21±1.31 gm, least disintegration time of 9±1.60 sec and highest drug release of 98.41±1.81% is chosen optimal formulation with maximum content uniformity and folding endurance. It is evident from the above results that the developed formulation can be an innovative dosage form to improve the drug delivery, quick onset of action as well as improve patient compliance in the effective management of depression.
ABSTRACT
We conducted a survey of the malaria vectors in an area where a power line had been constructed, between the municipalities of Porto Velho and Rio Branco, in the states of Rondônia and Acre, respectively. The present paper relates to the results of the survey of Anopheles fauna conducted in the state of Rondônia. Mosquito field collections were performed in six villages along the federal highway BR 364 in the municipality of Porto Velho, namely Porto Velho, Jaci Paraná, Mutum Paraná, Vila Abunã, Vista Alegre do Abunã, and Extrema. Mosquito captures were performed at three distinct sites in each locality during the months of February, July, and October 2011 using a protected human-landing catch method; outdoor and indoor captures were conducted simultaneously at each site for six hours. In the six sampled areas, we captured 2,185 mosquitoes belonging to seven Anopheles species. Of these specimens, 95.1% consisted of Anopheles darlingi, 1.8% An. triannulatus l.s., 1.7% An. deaneorum, 0.8% An. konderi l.s., 0.4 An. braziliensis, 0.1% An. albitarsis l.s., and 0.1% An. benarrochi. An. darlingi was the only species found in all localities; the remaining species occurred in sites with specific characteristics.
Foi realizado levantamento de vetores de malária na área que compreende a construção da linha de transmissão entre os municípios de Porto Velho e Rio Branco, estados de Rondônia e Acre, respectivamente. Os dados aqui apresentados mostram os resultados do levantamento da fauna dos Anopheles realizado em Rondônia. As capturas foram realizadas no município de Porto Velho em seis aglomerados populacionais ao longo da rodovia federal BR 364, denominados Porto Velho, Jaci Paraná, Mutum Paraná, Vila Abunã, Vista Alegre do Abunã e Extrema. As capturas ocorreram em três diferentes pontos de cada uma das localidades nos meses de fevereiro, julho e outubro de 2011, seguindo a metodologia de coleta por atração humana protegida em dois ambientes, sendo no intradomicílio e no peridomicílio simultaneamente com duração de seis horas. Nas áreas amostradas foram capturados 2.185 anofelinos pertencentes a sete espécies de Anopheles sp. dos quais 95,1% foram identificados como Anophels darlingi, 1,8% An. triannulatus l.s., 1,7% An. deaneorum, 0,8% An. konderi l.s., 0,4 An. braziliensis, 0,1% An. albitarsis l.s., e 0,1% An. benarrochi. Anopheles darlingi foi a única espécie amostrada em todas as localidades enquanto as demais espécies, ocorreram em locais com características singulares.
Subject(s)
Animals , Humans , Anopheles/classification , Insect Vectors/classification , Brazil , Malaria/transmission , Population Density , Rural Population , SeasonsABSTRACT
Object To investigate the effects of Heart-benefitting Recipe (HBR) on the histopathological changes of Alzheimer's disease (AD) rats after unilateral amyloid-?_ 25-35 protein (A?_ 25-35 ) injection into the amygdala. Methods The experimental rat models with dementia,spatial learning,and memory impairment were induced by unilateral amyloid ?_ 25-35 protein (A?_ 25-35 ) injection into the amygdala of rats. The spatial learning and memory ability and the function of cholinergic system were observed by the means of Morris water maze and radioligand binding assay. The protein expression of A?_ 1-40 ,AT_8 was estimated with immunohistochemistry. The expression of mRNA related to APP was observed by RT-PCR. Results HBR significantly alleviated the spatial learning and memory impairment induced by A?_ 25-35 and remarkably increased the activity of ChAT and Rt of M-receptor binding sites of these models. The expressions of A?_ 1-40 protein and APP mRNA in cortex and hippocampus of AD rats were decreased remarkably by HBR. The results of immunohistochemistry showed that the expression of hyperphosphorilated tau AT_ 8 in cortex and hippocampus of AD rats was inhibited significantly by HBR. Conclusion HBR can effectively improve the spatial learning and memory impairment,decrease the deposition of A?_ 1-40 ,and alleviate the hyperphos-phorylation of tau of AD rats induced by A?_ 25-35 .