995 resultados para eumenine mastoparan AF


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Eight kinds of pigment-protein complexes were resolved from the thylakoid membrane of the brown alga (Undaria pinnatifida Harv.) by using non-ionic detergent decanoyl-N-methylglucamide and PAGE technique. According to the apparent molecular weights, spectra characteristics, polypeptide compositions and referring to the higher plant spinach, eight pigment-protein complexes were named under Anderson's terminology system as CP I a, CP I, CPa, LHC1, LHC2, LHC3, LHC4, LHC5.

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Suspended particulate matter (SPM) measurements obtained along a cross-section in the central English Channel (Wight-Cotentin transect) indicate that the area may be differentiated into: (1) an English coastal zone, associated with the highest concentrations; (2) a French coastal zone, with intermediate concentrations; and (3) the offshore waters of the Channel, characterised by a very low suspended-sediment load. The SPM particle-size distribution was modal close to the English coast (main mode 10-12 mu m); the remainder of the area was characterised by flat SPM distributions. Examination of the diatom communities in the SPM suggest:; that material resuspended in the intertidal zone and the estuarine environments was advected towards the offshore waters of the English Channel. Considerable variations in SPM concentrations occurred during a tidal cycle: maximum concentrations were sometimes up to 3 times higher than the minimum concentrations, Empirical orthogonal function (EOF) analysis of the SPM concentration time series indicates that, although the bottom waters were more turbid than the surficial waters, this was not likely to be the result of in situ sediment resuspension. Instead, the observed variations appear to be controlled mainly by advective mechanisms. The limited resuspension was probably caused by: (1) the limited availability of fine-grained material within the bottom sediments, and (2) 'bed-armouring' processes which protect the finer-grained fractions of the seabed material from erosion and entrainment within the overlying flow during the less energetic stages of the tide.

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本研究应用显带技术和荧光原位杂交(Fluorescence in situ hybridization,FISH)技术,鉴定了牡蛎的染色体;应用FISH方法定位了一系列的重复序列和大分子的P1克隆DNA;制备了染色体特异性探针。应用FISH特异性探针成功地鉴定了长牡蛎的三体10。结果如下:1.分析了G带和C带在美洲牡蛎染色体上的分布。G带在每一条染色体上的带型不同,某些染色体间(如第1对和第4对染色体,第7对和第9对染色体)的带型差别不是很明显。G带型容易受染色体收缩程度的影响。C带型重复性较好,染色体带型较清楚,分布在染色体的端粒区域和着丝粒区域。G带和C带带型能够用来鉴定牡蛎的染色体,但是重复性低和带型差异不显著,并不适合常规的染色体鉴定。2.早期胚胎和担轮幼虫制备的染色体适合于FISH分析。染色体制备方法重复性好,可适用于其它贝类的染色体制备。3.研究了重复序列基因--rDNA的定位:1)18S-5.8S rDNA在研究的五种巨蛎属Crassostrea牡蛎均只有一个位 点。太平洋种(C.gigas,C. ariakensis和C. plicatula)中,杂交信号位于最短的染色体一第10对染色体长臂的端粒区域,在大西洋种(C. virginica和C. rhizophorae)中,同一序列定位在第2对染色体短臂的端粒区域。2)18S-28S rDNA在两种蛤中有两个位点。rDNA探针定位在侏儒蛤(Mulinis Lateralis)的第15对和第19对染色体的端粒区域,同一序列定位在硬壳蛤(Mercenaria mercenaria)的第10对染色体的长臂和第12对染色体短臂的端粒区域。信号强度在两对染色体之间有差异。 3)5s rDNA位于美洲牡蛎的第5对染色体的短臂上靠近着丝粒区域和第6 对染色体的短臂的中间区域。信号强度在两对染色体之间没有显著差异。5S rDNA探针可以作为鉴定和识别第5对和第6对染色体的特异性探针。4.研究了一些重复序列的定位1)两个短的重复序列1G8,1P2均产生很强的荧光信号分布在美洲牡蛎所有的染色体上。在低严谨条件下,这些序列均产生很强的信号散布在所有的染色体上。在高严谨条件下,信号强度大大减弱,但是信号仍散布在所有的染色体上。这些重复序列散布在美洲牡蛎的整个基因组中。2)高度重复序列Cgl70产生的信号分布在长牡蛎的7对染色体的着丝粒区域,没有发现间区信号。在第1对,第2对,第4对和第7对染色体上的荧光信号强且稳定。在第5对,第8对和第10对染色体上的信号相对弱且不稳定。在剩余的染色体上(第3对,第6对和第9对染色体)没有检测到荧光信号。结果表明此卫星序列是一个着丝粒卫星序列。在美洲牡蛎的染色体上没有检测到荧光信号,表明了这个着丝粒卫星序列在这两种牡蛎中的分布存在着显著的差异。3)脊椎动物端粒序列(TTAGGG)n的FISH信号局限在四种双壳贝类(美洲牡蛎,the mangrove oyster,硬壳蛤,侏儒蛤)所有染色体的端粒区域,没有发现间区信号的存在。研究结果与已报道的研究结果表明脊椎动物端粒序列或许存在于所有双壳贝类的染色体末端。双壳贝类是目前研究过的唯一含有脊椎动物端粒序列DNA的无脊椎动物。4)研究了RAPD探针在美洲牡蛎染色体上的定位。大多数RAPD探针产生了多个信号散布在间期细胞核和所有的染色体上。引物OPX-03,OPX-04,OPX—06,OPG-02,OPM—04,OPM-11,0PS-02制备的探针在适宜的条件下产生特异性荧光 信号,分布在牡蛎的特定的染色体上。PCR特异性带产生的探针OPX—06—310和0PG-02—300产生了特异性的荧光信号:OPX—06—310产生的信号位于第5对染色体的短臂的近端粒区域,0PG—02—300探针定位到第3对染色体的短臂上。这两个探针是鉴定美洲牡蛎单条染色体的特异性探针。5.研究了大分子Pl克隆DNA(插入片断为80~100 kb)在美洲牡蛎染色体上的定位。Pl克隆DNA通过切口平移方法标记digoxigenin—11-dUTP用作FISH的探针。Cot-1 DNA作为竞争剂有效地抑制了Pl克隆序列中的重复序列产生的信号。杂交信号用fluorescein标记的anti—digoxigenin抗体来检测,用两层抗体rabbit-anti-sheep抗体和FITC anti—rabbit抗体来扩增信号。9个P1探针成功地定位在特定的染色体上。46—1探针杂交到第1对染色体的长臂靠近着丝粒区域;47-10探针定位到第2对染色体的长臂近端粒区域;Cvpl和48-13两探针定位到第3对染色体上:Cvpl位于短臂的端粒区域,48-13探针位于长臂的近着丝粒区域;48—10探针杂交到第4对染色体的长臂上;48-1探针杂交到第5对染色体长臂的近着丝粒区域;49-11探针位于第7对染色体长臂上;探针49-10和44-11位于第8对染色体长臂上。同时我们成功地将2个P1探针杂交到同一染色体分裂相中,进一步确定了Pl探针在美洲牡蛎染色体 上的定位。6.应用18S-28S rDNA探针成功地鉴定出长牡蛎非整倍体中的三体10。经鉴定AF-35,AF-39和AF-3三体家系属于三体10家系。rDNA探针分布在三条染色体上,即多出的一条染色体为染色体10。相应地在间期细胞核上有三个信号出现。AF-34和AF-36家系不属于三体10家系。rDNA探针分布在两条染色体上,相应地在间期细胞核上有两个信号出现。FISH和染色体特异性探针为非整倍体的鉴定提供了一个快速准确可靠的方法和途径。

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近年来,世界沿海国家有害赤潮发生的频率、规模及危害都有上升趋势,有害赤潮已经成为重要的近海环境问题之一。要有效防范有害赤潮带来的危害效应,建立和发展可靠、有效的赤潮监测手段非常重要。目前,对于赤潮藻种的监测主要依靠显微观察的方法,在实际应用中经常遇到困难。首先,亲缘关系相近的物种在形态上差异很小,如甲藻门亚历山大藻属的一些种类,仅细胞壁上个别甲片的结构有细微差别,并且这些形态学指标还容易受环境条件及生长阶段的影响。另外,这种以形态学为基础的分析方法,分析速度慢、耗时长,对操作人员的要求较高,难以满足浮游植物种群动力学监测“量大、连续”的要求。因此,本研究将分子生物学的技术和方法应用于赤潮监测,力求提高赤潮藻种鉴定的准确性和检测工作的效率。 亚历山大藻是一类重要的有害赤潮藻,该藻属中一些产毒特性差别很大的藻种,单从表形特征难以明确区分,从而限制了基于形态观察的监测技术的应用。本研究中,我们尝试应用分子生物学技术与方法,开展了该藻属藻种分子鉴定和荧光原位杂交检测方法的研究。在亚历山大藻的分子鉴定方面,我们采用了核糖体RNA基因(rDNA)序列分析的方法,首次测定了9株分离自中国沿海的(以及实验室保有的其它两株)亚历山大藻的rDNA序列全长,其中包括核糖体小亚基(SSU)rDNA、大亚基(LSU)rDNA、5.8S rDNA及内转录间隔区(ITS)区序列。序列分析结果显示,这些藻株包含了5种核糖体类型,分别是塔玛复合种亚洲温带(Temperate Asian)核糖体类型(TSC-TA),塔玛复合种西欧(West European)核糖体类型(TSC-WE),相关亚历山大藻(A. affine)核糖体类型(AF),微小亚历山大藻(A. minutum)葡萄牙(Portugal)核糖体类型(M-PO)和微小亚历山大藻新西兰(New Zealand)核糖体类型(M-NZ)。将测获的rDNA序列划分为若干保守性不同的区段,分别进行系统发育分析(结合GenBank数据库中保存的其它亚历山大藻相关序列)。结果显示,LSU rDNA D1-D2区是对该藻属藻种进行分子鉴定和系统发育研究的较好区段。同时,为解决建立亚历山大藻克隆培养的困难,我们应用单细胞rDNA序列分析方法,对亚历山大藻单个细胞直接进行了种类鉴定。结果表明,该方法适用于不同生活史阶段的亚历山大藻。 在亚历山大藻的检测技术方面,我们进一步扩展和完善了针对完整细胞的荧光原位杂交检测方法。首先,通过对不同核糖体类型藻株rDNA序列信息的对比分析,针对各自特异的序列位点,设计了特异性rRNA标记探针。经荧光原位杂交实验检验,实现了对5种核糖体类型亚历山大藻的特异性标记。其中,针对WE、M-PO及M-NZ核糖体型的特异性探针为首次获得,另外两个探针是针对TA和AF核糖体类型rRNA新的位点所设计。同时,对影响探针标记效果的诸多因素进行了分析和探讨。此外,在2007年春季长江口海域赤潮调查中,首次应用特异性核酸探针和荧光原位杂交检测方法,调查了该海域亚历山大藻的丰度。结果表明,在4月4日-4月10日的样品中,亚历山大藻达到了较高的密度,最高密度达到103cells/L。同时发现,实验中样品的保存方法有待改进。随后的研究表明,盐醇固定方法及多聚甲醛/甲醇固定方法,可以较好的保持rRNA不被降解并适宜杂交(至少3个月时间)。 总之,本研究首次测定并分析了11株亚历山大藻(9株分离自中国沿海)的rDNA全序列信息。在此基础上,获得了5种核糖体类型亚历山大藻的特异性rRNA标记探针,其中3种为首次获得。另外,实验证明,单细胞rDNA分析技术和荧光原位杂交检测方法,在自然水体中亚历山大藻的直接鉴定及丰度调查中,均具有良好的应用前景。这一工作为我国近海亚历山大藻的鉴定和检测提供了理论依据和方法学基础,希望对该藻赤潮的监测工作有推动作用。 关键词:亚历山大藻 遗传探针 rRNA rDNA 荧光原位杂交 系统发育

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A method based on protein phosphatase enzyme activity inhibition for the detection of diarrhetic shellfish poison (DSP) was used to analyze the DSP toxicity in three oyster samples. Based on the standard dose-effect curve developed with a series of okadaic acid (OA) standard solutions, the DSP toxicity of the three oyster samples collected were screened, and the results showed that there were no OA and dinophysis toxins ( DTXs) in the samples without hydrolization. However, the OA toxicity could be detected in two of the hydrolyzed samples, and the OA toxicity of the two samples were 1.81 and 1.21 mu g OA eq./kg oyster, respectively.

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[目的]探索从虎耳草科植物中提取DNA的有效方法.[方法]采用改进的CTAB法,从11种虎耳草科植物中提取DNA.以提取的DNA为模板,利用通用引物"psb AF"和"trnHR"对虎耳草科植物叶绿体DNA psb A-trn H片段进行PCR扩增.[结果]通过该方法提取的DNA纯度较高,质量较好.用所得DNA进行psb A-trn H扩增的产量高,可用于后续的测序等分析.对山地虎耳草的PCR产物纯化后进行测序,得到262 bp的序列.将其与CenBank中的虎耳草属其他植物的psb A-tm H序列进行比对分析,证实该序列为目标psb A-trn H片段的区域.[结论]该方法可有效去除次生物质对DNA的干扰,提取的基因组DNA可用于叶绿体psb A-trn H测序分析和其他遗传学分析.

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根据K—Ar法和Ar-Ar法定年数据,藏北新生代火山岩从早到晚可划分为多格错仁和走构油茶错高钾钙碱性系列(40—30Ma)、巴毛穷宗和鱼鳞山白榴石碧玄岩—响岩系列(29—24Ma)以及黑石北湖钾玄岩系列(1.5Ma)。从岩石化学和同位素方面对区内新生代火山岩进行了研究,认为藏北新生代火山岩同时具有板内碱性玄武岩和岛弧玄武岩的双重地球化学特征。在成因和源区上各系列存在着差异,高钾钙碱性系列为富集地幔玄武岩底侵作用导致地壳物质部分熔融的产物,并经历了地壳的AF过程。而白榴石碧玄岩—响岩系列和钾玄岩系列来自与古俯冲有关的“古老富集地幔”,为岩浆分离结晶作用的产物。岩浆起源深度随时间变新逐渐变大。

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Este trabalho teve por objetivos avaliar combinações enxerto/porta-enxerto e modalidades de enxertia no sentido de indução de resistência em algumas espécies de meliáceas à broca causada pela Hypsipyla grandella. Foram testadas modalidades de enxertia e combinações intraespecíficas, intra e intergenéricas e interfamílias. A combinação toona/mogno, métodos GFMT e GFCT obtiveram, respectivamente o segundo e o terceiro pegamento médio, diferindo estatisticamente das demais. As combinações mogno/ toona e toona/mogno apresentaram bons resultados de pegamento na enxertia. Entretanto, as plantas definharam e acabaram morrendo, restando vivas as combinações mogno/mogno (testemunha). O melhor pegamento foi obtido na combinação cedro branco/cinamomo, diferindo significativamente das demais. A combinação cedro branco/toona apresentou o segundo melhor pegamento, diferindo significativamente das demais. As combinações MBr/Af ? GFMT, MAf/MBR ? GFMT e MBr/Af ? Borbulhia em T invertido apresentaram o terceiro melhor pegamento. O comportamento das melhores combinações foi avaliado em condições de campo. Os resultados mostraram incompatibilidade enxerto/porta-enxerto na fase de enxertia para todas as combinações testadas, à exceção das combinações mogno/mogno africano, mogno/mogno (testemunha) e cedro branco/cedro australiano. Em condições de campo, a combinação mogno/ mogno africano foi atacada pela H. grandella (apresentou sintomas de danos da broca).

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A recent meta-analysis by Iskandar et al. (1) nicely showed that endurance athletes have larger left atrial (LA) diameters compared with control subjects. Yet only 9 of 54 studies included in their analysis reported LA volume values corrected for body surface area (BSA). In fact, few studies have determined LA volume in young athletes, and, to the best of our knowledge, no study has reported this variable in older athletes. This is an important question given the growing debate about the potential deleterious effects of long-term strenuous endurance exercise on the human heart, notably the higher risk of atrial fibrillation (AF), a condition for which both atrial dilation and the normal aging process are thought to be potential causative mechanisms (2). Thus, we aimed to assess the long-term consequences of endurance exercise on LA volume in athletes who were highly competitive at younger ages and are still active. To this end, we compared BSA-corrected LA volumes determined with late gadolinium enhancement magnetic resonance imaging (LGE-MRI) in former elite endurance athletes and sedentary control subjects.

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How rainfall infiltration rate and soil hydrological characteristics develop over time under forests of different ages in temperate regions is poorly understood. In this study, infiltration rate and soil hydrological characteristics were investigated under forests of different ages and under grassland. Soil hydraulic characteristics were measured at different scales under a 250 year old grazed grassland (GL), a six (6 yr) and 48 (48 yr) year old Scots pine (Pinus sylvestris) plantation, remnant 300 year old individual Scots pines (OT) and a 4000 year old Caledonian Forest (AF). In-situ field saturated hydraulic conductivity (Kfs) was measured and visible root:soil area was estimated from soil pits. Macroporosity, pore structure, and macropore connectivity were estimated from X-ray tomography of soil cores, and from water-release characteristics. At all scales the median values for Kfs, root fraction, macro-porosity and connectivity values tended to AF > OT > 48 yr > GL > 6 yr, indicating that infiltration rates and water storage increased with forest age. The remnant Caledonian Forest had a huge range of Kfs (12 to > 4922 mm h-1), with maximum Kfs values 7 to 15 times larger than 48-year-old Scots pine plantation, suggesting that undisturbed old forests, with high rainfall and minimal evapotranspiration in winter, may act as important areas for water storage and sinks for storm rainfall to infiltrate and transport to deeper soil layers via preferential flow. The importance of the development of soil hydrological characteristics under different aged forests is discussed.

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BACKGROUND:Cardiovascular disease (CVD) and its most common manifestations - including coronary heart disease (CHD), stroke, heart failure (HF), and atrial fibrillation (AF) - are major causes of morbidity and mortality. In many industrialized countries, cardiovascular disease (CVD) claims more lives each year than any other disease. Heart disease and stroke are the first and third leading causes of death in the United States. Prior investigations have reported several single gene variants associated with CHD, stroke, HF, and AF. We report a community-based genome-wide association study of major CVD outcomes.METHODS:In 1345 Framingham Heart Study participants from the largest 310 pedigrees (54% women, mean age 33 years at entry), we analyzed associations of 70,987 qualifying SNPs (Affymetrix 100K GeneChip) to four major CVD outcomes: major atherosclerotic CVD (n = 142; myocardial infarction, stroke, CHD death), major CHD (n = 118; myocardial infarction, CHD death), AF (n = 151), and HF (n = 73). Participants free of the condition at entry were included in proportional hazards models. We analyzed model-based deviance residuals using generalized estimating equations to test associations between SNP genotypes and traits in additive genetic models restricted to autosomal SNPs with minor allele frequency [greater than or equal to]0.10, genotype call rate [greater than or equal to]0.80, and Hardy-Weinberg equilibrium p-value [greater than or equal to] 0.001.RESULTS:Six associations yielded p <10-5. The lowest p-values for each CVD trait were as follows: major CVD, rs499818, p = 6.6 x 10-6; major CHD, rs2549513, p = 9.7 x 10-6; AF, rs958546, p = 4.8 x 10-6; HF: rs740363, p = 8.8 x 10-6. Of note, we found associations of a 13 Kb region on chromosome 9p21 with major CVD (p 1.7 - 1.9 x 10-5) and major CHD (p 2.5 - 3.5 x 10-4) that confirm associations with CHD in two recently reported genome-wide association studies. Also, rs10501920 in CNTN5 was associated with AF (p = 9.4 x 10-6) and HF (p = 1.2 x 10-4). Complete results for these phenotypes can be found at the dbgap website http://www.ncbi.nlm.nih.gov/projects/gap/cgi-bin/study.cgi?id=phs000007.CONCLUSION:No association attained genome-wide significance, but several intriguing findings emerged. Notably, we replicated associations of chromosome 9p21 with major CVD. Additional studies are needed to validate these results. Finding genetic variants associated with CVD may point to novel disease pathways and identify potential targeted preventive therapies.

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A female patient, with normal familial history, developed at the age of 30 months an episode of diarrhoea, vomiting and lethargy which resolved spontaneously. At the age of 3 years, the patient re-iterated vomiting, was sub-febrile and hypoglycemic, fell into coma, developed seizures and sequels involving right hemi-body. Urinary excretion of hexanoylglycine and suberylglycine was low during this metabolic decompensation. A study of pre- and post-prandial blood glucose and ketones over a period of 24 hours showed a normal glycaemic cycle but a failure to form ketones after 12 hours fasting, suggesting a mitochondrial β-oxidation defect. Total blood carnitine was lowered with unesterified carnitine being half of the lowest control value. A diagnosis of mild MCAD deficiency (MCADD) was based on rates of 1-14C-octanoate and 9, 10-3H-myristate oxidation and of octanoyl-CoA dehydrogenase being reduced to 25% of control values. Other mitochondrial fatty acid oxidation proteins were functionally normal. De novo acylcarnitine synthesis in whole blood samples incubated with deuterated palmitate was also typical of MCADD. Genetic studies showed that the patient was compound heterozygous with a sequence variation in both of the two ACADM alleles; one had the common c.985A>G mutation and the other had a novel c.145C>G mutation. This is the first report for the ACADM gene c.145C>G mutation: it is located in exon 3 and causes a replacement of glutamine to glutamate at position 24 of the mature protein (Q24E). Associated with heterozygosity for c.985A>G mutation, this mutation is responsible for a mild MCADD phenotype along with a clinical story corroborating the emerging literature view that patients with genotypes representing mild MCADD (high residual enzyme activity and low urinary levels of glycine conjugates), similar to some of the mild MCADDs detected by MS/MS newborn screening, may be at risk for disease presentation.

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BACKGROUND: Methicillin-resistant Staphylococcus aureus (MRSA) is a common cause of complicated skin and skin-structure infection (cSSSI). Increasing antimicrobial resistance in cSSSI has led to a need for new safe and effective therapies. Ceftaroline was evaluated as treatment for cSSSI in 2 identical phase 3 clinical trials, the pooled analysis of which is presented here. The primary objective of each trial was to determine the noninferiority of the clinical cure rate achieved with ceftaroline monotherapy, compared with that achieved with vancomycin plus aztreonam combination therapy, in the clinically evaluable (CE) and modified intent-to-treat (MITT) patient populations. METHODS: Adult patients with cSSSI requiring intravenous therapy received ceftaroline (600 mg every 12 h) or vancomycin plus aztreonam (1 g each every 12 h) for 5-14 days. RESULTS: Of 1378 patients enrolled in both trials, 693 received ceftaroline and 685 received vancomycin plus aztreonam. Baseline characteristics of the treatment groups were comparable. Clinical cure rates were similar for ceftaroline and vancomycin plus aztreonam in the CE (91.6% vs 92.7%) and MITT (85.9% vs 85.5%) populations, respectively, as well as in patients infected with MRSA (93.4% vs 94.3%). The rates of adverse events, discontinuations because of an adverse event, serious adverse events, and death also were similar between treatment groups. CONCLUSIONS: Ceftaroline achieved high clinical cure rates, was efficacious against cSSSI caused by MRSA and other common cSSSI pathogens, and was well tolerated, with a safety profile consistent with the cephalosporin class. Ceftaroline has the potential to provide a monotherapy alternative for the treatment of cSSSI. TRIAL REGISTRATION: ClinicalTrials.gov identifiers: NCT00424190 for CANVAS 1 and NCT00423657 for CANVAS 2.

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Transient overexpression of defined combinations of master regulator genes can effectively induce cellular reprogramming: the acquisition of an alternative predicted phenotype from a differentiated cell lineage. This can be of particular importance in cardiac regenerative medicine wherein the heart lacks the capacity to heal itself, but simultaneously contains a large pool of fibroblasts. In this study we determined the cardio-inducing capacity of ten transcription factors to actuate cellular reprogramming of mouse embryonic fibroblasts into cardiomyocyte-like cells. Overexpression of transcription factors MYOCD and SRF alone or in conjunction with Mesp1 and SMARCD3 enhanced the basal but necessary cardio-inducing effect of the previously reported GATA4, TBX5, and MEF2C. In particular, combinations of five or seven transcription factors enhanced the activation of cardiac reporter vectors, and induced an upregulation of cardiac-specific genes. Global gene expression analysis also demonstrated a significantly greater cardio-inducing effect when the transcription factors MYOCD and SRF were used. Detection of cross-striated cells was highly dependent on the cell culture conditions and was enhanced by the addition of valproic acid and JAK inhibitor. Although we detected Ca(2+) transient oscillations in the reprogrammed cells, we did not detect significant changes in resting membrane potential or spontaneously contracting cells. This study further elucidates the cardio-inducing effect of the transcriptional networks involved in cardiac cellular reprogramming, contributing to the ongoing rational design of a robust protocol required for cardiac regenerative therapies.