Identification of cardiac stem cells within mature cardiac myocytes

Cardiac stem cells are described in a number of mammalian species including humans. Cardiac stem cell clusters consisting of both lineage-negative and partially committed cells are generally identified between contracting cardiac myocytes. In the present study, c-kit⁺, Sca⁺, and Isl1⁺ stem cells were revealed to be located inside the sarcoplasm of cardiac myocytes in myocardial cell cultures derived from newborn, 20-, and 40-day-old rats. Intracellularly localized cardiac stem cells had a coating or capsule with a few pores that opened into the host cell sarcoplasm. The similar structures were also identified in the suspension of freshly isolated myocardial cells (ex vivo) of 20- and 40-day-old rats. The results from this study provide direct evidence for the replicative division of encapsulated stem cells, followed by their partial cardiomyogenic differentiation. The latter is substantiated by the release of multiple transient amplifying cells following the capsule rupture. In conclusion, functional cardiac stem cells can reside not only exterior to but also within cardiomyocytes.     

Molecular phylogenetic analysis shows that <Emphasis Type="Italic">Amanita ponderosa</Emphasis> and <Emphasis Type="Italic">A. curtipes</Emphasis> are distinct species

Amanita curtipes and A. ponderosa are two Mediterranean taxa sharing a number of morphological features as well as their habitat. Their synonymy or variety status has been proposed by several authors. To clarify this taxonomic issue we have sequenced the D1-D2 domains of the 28S rRNA gene as well as the complete ITS1-5.8S-ITS2 region of several specimens of the two species collected in Spain, and aligned these sequences with those from other Amanita species. Molecular phylogenetic analysis based on the two regions revealed that A. ponderosa and A. curtipes are clearly distinct species. The distribution of Amanita species in the phylogenetic trees was consistent with the division of the genus in subgenera and sections as proposed by previous authors. Sequences of A. ponderosa and A. curtipes were grouped in a monophyletic cluster together with other species of the section Amidella. However, A. ponderosa was closer to other species in the section, such as A. peckiana and A. volvata, than to A. curtipes. We also indicate the macromorphological characters that are most useful to reliably distinguish A. ponderosa and A. curtipes.     

培养人皮下脂肪干细胞纵向分化的实验研究

目的:建立人脂肪干细胞(adipose-derived stem cells,ADSCs)分离、培养的方法,观察其生物学特性,探讨其纵向分化的能力.方法:自皮下脂肪组织获得梭形细胞,观察细胞生物学特征,免疫组化鉴定波形蛋白和CD44.以含有胰岛素、地塞米松、1.甲基-3-异丁基-黄嘌呤的无血清混合培养基诱导其向脂肪细胞纵向分化,以细胞形态学变化.油红O染色和测定甘油磷酸脱氢酶活性判定分化是否成功.结果:人皮下脂肪中能够分离培养出生长旺盛的脂肪干细胞,诱导培养7d后细胞由梭形逐渐变圆,胞质内出现脂滴后逐渐增多融合为脂泡,并与甘油磷酸脱氢酶活性变化相吻合.油红O染色显示约(78.6±2.2)%细胞转变为脂肪细胞.结论:人皮下脂肪中分离的脂肪干细胞在体外诱导条件下能纵向分化为脂肪细胞,可能成为修复软组织缺损及整形美容的又一干细胞来源.     

吉林蛟河针阔混交林树木生长的空间关联格局

以吉林蛟河21.12hm2(660m×320m)针阔混交林样地为对象,利用2009年和2014年森林生长观测数据,研究树木生长的空间自相关格局及其生境影响机制。在样地生境型划分结果的基础上,采用Ripley's L(r)函数分析不同生境型中树木种群空间分布特征;利用标记相关函数分析不同生境型中树木生长特征的空间关联格局。研究结果表明:(1)红松(生境型3:1—5m)、蒙古栎(生境型3:1—3m)、胡桃楸(生境型2:1—2m;生境型3:1—7m)、黄檗(生境型2:1—3m;生境型4:1—5m)、水曲柳(生境型3:1—2m;生境型4:1—2m)、瘤枝卫矛(生境型2:1—15m)在特定生境和空间尺度上呈随机分布,但空间格局仍以聚集性分布为主;其余10个物种则在全部0—30m尺度上呈聚集分布。(2)标记相关函数分析显示春榆、毛榛、色木槭、瘤枝卫矛和千金榆的径向生长至少在一个生境中表现出正相关格局;暴马丁香、胡桃楸、裂叶榆、瘤枝卫矛、水曲柳、紫椴、糠椴、毛榛、色木槭和白牛槭的径向生长至少在一个生境中表现出负相关格局;红松、黄檗、蒙古栎和簇毛槭的径向生长在全部尺度上均未检测到显著的空间关联格局。因此,不同树种径向生长的空间自相关特征不同,树种生长特征的空间关联格局具有明显的生境依赖性。     

The bromodomain inhibitor JQ1 triggers growth arrest and apoptosis in testicular germ cell tumours in vitro and in vivo

Type II testicular germ cell cancers (TGCT) are the most frequently diagnosed tumours in young men (20–40 years) and are classified as seminoma or non‐seminoma. TGCTs are commonly treated by orchiectomy and chemo‐ or radiotherapy. However, a subset of metastatic non‐seminomas (embryonal carcinomas) displays only incomplete remission or relapse and requires novel treatment options. Recent studies have shown effective application of the small‐molecule inhibitor JQ1 in tumour therapy, which interferes with the function of ‘bromodomain and extraterminal (BET)’ proteins. JQ1‐treated TGCT cell lines display up‐regulation of genes indicative for DNA damage and cellular stress response and induce cell cycle arrest. Embryonal carcinoma (EC) cell lines, which presented as JQ1 sensitive, display down‐regulation of pluripotency factors and induction of mesodermal differentiation. In contrast, seminoma‐like TCam‐2 cells tolerated higher JQ1 concentrations and were resistant to differentiation. ECs xenografted in vivo showed a reduction in tumour size, proliferation rate and angiogenesis in response to JQ1. Finally, the combination of JQ1 and the histone deacetylase inhibitor romidepsin allowed for lower doses and less frequent application, compared with monotherapy. Thus, we propose that JQ1 in combination with romidepsin may serve as a novel therapeutic option for (mixed) TGCTs.     

Phenotypic assays for analyses of pluripotent stem cell–derived cardiomyocytes

Stem cell–derived cardiomyocytes (CMs) hold great hopes for myocardium regeneration because of their ability to produce functional cardiac cells in large quantities. They also hold promise in dissecting the molecular principles involved in heart diseases and also in drug development, owing to their ability to model the diseases using patient‐specific human pluripotent stem cell (hPSC)–derived CMs. The CM properties essential for the desired applications are frequently evaluated through morphologic and genotypic screenings. Even though these characterizations are necessary, they cannot in principle guarantee the CM functionality and their drug response. The CM functional characteristics can be quantified by phenotype assays, including electrophysiological, optical, and/or mechanical approaches implemented in the past decades, especially when used to investigate responses of the CMs to known stimuli (eg, adrenergic stimulation). Such methods can be used to indirectly determine the electrochemomechanics of the cardiac excitation‐contraction coupling, which determines important functional properties of the hPSC‐derived CMs, such as their differentiation efficacy, their maturation level, and their functionality. In this work, we aim to systematically review the techniques and methodologies implemented in the phenotype characterization of hPSC‐derived CMs. Further, we introduce a novel approach combining atomic force microscopy, fluorescent microscopy, and external electrophysiology through microelectrode arrays. We demonstrate that this novel method can be used to gain unique information on the complex excitation‐contraction coupling dynamics of the hPSC‐derived CMs.     

Probing disorders of the nervous system using reprogramming approaches

The groundbreaking technologies of induced pluripotency and lineage conversion have generated a genuine opportunity to address fundamental aspects of the diseases that affect the nervous system. These approaches have granted us unrestricted access to the brain and spinal cord of patients and have allowed for the study of disease in the context of human cells, expressing physiological levels of proteins and under each patient's unique genetic constellation. Along with this unprecedented opportunity have come significant challenges, particularly in relation to patient variability, experimental design and data interpretation. Nevertheless, significant progress has been achieved over the past few years both in our ability to create the various neural subtypes that comprise the nervous system and in our efforts to develop cellular models of disease that recapitulate clinical findings identified in patients. In this Review, we present tables listing the various human neural cell types that can be generated and the neurological disease modeling studies that have been reported, describe the current state of the field, highlight important breakthroughs and discuss the next steps and future challenges.     

Systematics of Pseudorchis albida s.l. (Orchidaceae) in Europe and North America

In Scandinavia Pseudorchis albida (Orchidaceae)usually divided into the lowland and subalpine P. albida s.s. and the more or less alpine P. straminea. There have been some uncertainties and conflicting views concerning die taxonomic treatment of diese taxa. To address this issue, herbarium specimens of P. albida s.l. were studied for variation in morphological characters. A small-scale population study approach was used, as herbarium sheets with two or three plants were used as population samples. Canonical Variates Analysis (CVA) indicated a distinction between taxa in population means, corresponding to P. albida s.s. and P. straminea , respectively. Principal Components Analysis (PCA), however, revealed an overlap between individuals of the two taxa. The PGA analysis, furthermore, revealed that the overlap was considerably larger in material from Central Europe man in material from Fennoscandia. Student t -tests on separate characters confirmed the picture, wim more characters significantly different in Fennoscandian than in Central European material. Furthermore, a Tukey-Kramer test revealed that there were small differences between regional populations of P. albida s.s. , while there were several significant differences in single characters between the Norm American regional population of P. straminea , as compared with the Central European and Fennoscandian regional populations. In Central Europe there is no clear separation between taxa, while in Fennoscandia the taxa are more clearly separated. This probably means that there is a difference in the time of establishment in the different regions. The author suggests a distinction of taxa at the subspecies level, and argues that the clear distinction seen in Fennoscandian material is due to separate immigration histories for die two subspecies into Fennoscandia after the last period of glaciation.