Development of a novel explant culture method for the isolation of mesenchymal stem cells from human breast tumor

Background: Mesenchymal stem cells (MSCs) were isolated from various sources, including various types of tumors. However choosing an appropriate isolation method is an important step in obtaining cells with optimal quality and yield in companion with economical considerations. The purpose of this study was to isolate more pure MSCs from human breast tumor tissue by a modified explant culture method.

Methods and Materials: The tumor tissues (n = 8) were cut into 1 to 3-mm cube-like pieces (explant). Each explant was placed in a well of 24-well format plates, cultured in Dulbecco’s Modified Eagle’s medium (DMEM), and maintained at 37°C with 5% humidified incubator. Morphological phenotypes of the cells were surveyed by an inverted microscope and wells with rather homogenous fibroblast-like morphology cell were considered as positive and selected for more expansion and characterization.

Results: A total of 185 wells, 63.7% of wells were positive that were chosen for expansion. Flowcytometry analysis demonstrated that isolated cells were positive for CD73, CD44, CD29, CD105, and CD90 but negative for CD11b, CD45, CD34, and HLA?DR. In addition, cells possessed the capability of multipotential differentiation into osteoblasts and adipocytes.     

Social environment alters central distribution of estrogen receptor alpha in juvenile prairie voles

It is well established that social environment, particularly isolation, has a significant impact on social behaviors and neuroendocrine responses. Estrogen receptor alpha (ERα) expression in limbic structures and associated nuclei is related to the display of social behaviors. We hypothesized that the stress of isolation would cause changes in the pattern of ERα expression in the brain. Using a highly social (typically monogamous and biparental) rodent species, the prairie vole (Microtus ochrogaster), we housed juvenile voles with a sibling, stranger or in isolation for either 4 days or 21 days. Housing manipulations began following weaning from parents and group housed siblings. Rodents may be especially sensitive to manipulations of their social environment during this juvenile period. In particular, female prairie voles are induced ovulators, reliant upon exposure to an unrelated male (male urine) to become reproductively active. ERα immunoreactivity was quantified in the medial preoptic area (MPOA), bed nucleus of the stria terminalis (BST), ventromedial nucleus of the hypothalamus (VMH) and medial amygdala (MeA). Significantly fewer ERα immunoreactive (ERα-ir) cells were labeled in the MPOA and BST of females isolated for 21 days compared with stranger housed females. Non-significant differences were shown in the VMH and MeA of females. No differences were found in voles isolated for 4 days. These results suggest that female prairie voles may be more sensitive than males to manipulations of their social environment during the juvenile period.     

Dimensionality of mate choice,sexual isolation,and speciation

Multiple cues, across multiple sensory modalities, are involved in mate choice in a wide range of animal taxa. This multiplicity leads to the prediction that, in adaptive radiations, sexual isolation results from divergence in multiple dimensions. However, difficulties in directly measuring preferences and detecting multiple effects limit our ability to empirically assess the number of independent traits contributing to mate choice and sexual isolation. We present an approach to estimate the dimensionality of sexual isolation using mating trials across groups of related populations. We analyze nine radiations: seven in fruit flies (Drosophila) and one each in salamanders (Desmognathus) and cichlid fishes (Pseudotropheus). We find strong evidence that multiple latent traits—linear combinations of phenotypic traits and preferences—are responsible for the patterns of sexual isolation in all nine radiations but that dimensionality has a strong upper limit. Just two latent traits are implicated in the majority of cases. Mapping along latent trait axes tests predictions of sexual-selection models and allows correlation with specific phenotypic traits and functional components of mate choice. We find support for the role of stabilizing natural selection on the sexually selected (male) traits. In the cichlids, latent-trait axes incorporate male-coloration patterns and exhibit convergence as well as divergence among populations. In the salamanders, temporal patterning in sensory modalities and male vs. female preferences are reflected in different latent-trait axes.     

Multicenter Analysis of Novel and Established Variables Associated with Successful Human Islet Isolation Outcomes

Islet transplantation is a promising therapy used to achieve glycometabolic control in a select subgroup of individuals with type I diabetes. However, features that characterize human islet isolation success prior to transplantation are not standardized and lack validation. We conducted a retrospective analysis of 806 isolation records from 14 pancreas‐processing laboratories, considering variables from relevant studies in the last 15 years. The outcome was defined as postpurification islet equivalent count, dichotomized into yields ≥315 000 or ≤220 000. Univariate analysis showed that donor cause of death and use of hormonal medications negatively influenced outcome. Conversely, pancreata from heavier donors and those containing elevated levels of surface fat positively influence outcome, as did heavier pancreata and donors with normal amylase levels. Multivariable logistic regression analysis identified the positive impact on outcome of surgically intact pancreata and donors with normal liver function, and confirmed that younger donors, increased body mass index, shorter cold ischemia times, no administration of fluid/electrolyte medications, absence of organ edema, use of University of Wisconsin preservation solution and a fatty pancreas improves outcome. In conclusion, this multicenter analysis highlights the importance of carefully reviewing all donor, pancreas and processing parameters prior to isolation and transplantation.     

Comparison of agar-based media for primary isolation of glycopeptide-resistant enterococci

Objective: To compare four vancomycin-containing agar media for the isolation of glycopeptide-resistant enterococci (GRE) from clinical fecal specimens: kanamycin-aesculin-azide (KAA) agar; bile-aesculin-polymixin (BAP) agar; aztreonam-amphotericin blood (CBAA) agar; and neomycin blood (CBN) agar.
Methods: Fecal specimens from 125 patients were inoculated onto each medium. Media were examined for enterococci after incubation for up to 48 h. Enterococci were identified to species level, and glycopeptide phenotypes were determined by measuring minimum inhibitory concentrations of vancomycin and teicoplanin.
Results: GRE were isolated from 44/125 samples. Enterococcus faecalis and Enterococcus faecium isolates, expressing glycopeptide resistance of the VanA or VanB phenotypes, were recovered from 27/33 (82%) specimens on BAP medium, 26/33 (79%) on KAA medium, and 21/33 (64%) on CBN and CBAA media. Enterococcus gallinarum and Enterococcus casseliflavus isolates expressing low-level glycopeptide resistance (VanC phenotype) were recovered from 14/15 (93%) specimens on CBAA medium, 7/15 (47%) on KAA and CBN media, and 6/15 (40%) on BAP medium.
Conclusions: The media tested in this study, with the exception of CBN medium, detected at least 75% of patients colonized by GRE. Further development of BAP, CBAA and KAA media is warranted to improve growth and selectivity.     

河北省虫媒病毒分离鉴定

目的 研究河北省蚊虫携带虫媒病毒情况。方法 在蚊虫孳生地采集标本液氯保存，按照20—30只／组经无菌处理后研磨，研磨液离心后接种C6／36细胞，连续三代观察致细胞病变情况；对细胞病变阳性的分离物，进行血清学和分子生物学鉴定。结果 在河北省涉县两个村共采集蚊虫1310只，分46组进行处理，共获得13株阳性分离物，其中两株分离物经间接免疫荧光检测提示为甲病毒属成员，用甲病毒属特异性引物RT-PCR扩增阳性，经核酸序列测定证实该序列与Getah病毒（AY702913．1）同源性最高（98％），初步鉴定为Getah病毒，其他病毒分离物尚在鉴定。结论 本研究从河北省蚊虫标本中分离到Getah病毒，这是我国内陆地区首次分离到该病毒。     

Populations of a Connecticut River midge structured by geological history and downstream gene flow

Connecticut River midges in the genus Axarus are constrained to semi-isolated populations associated with submerged clay exposures, and gene flow between populations is restricted. I sampled larval midges from fifteen spatially separated sites in the Connecticut River and determined karyotypes for 3111 larvae in two closely related species. In one of these species, an undescribed species referred to as Axarus species varvestris, 5chromosomal rearrangements were present as polymorphisms. Four of these are paracentric inversions, and the fifth is a terminal insertion or deletion. I used chromosomal rearrangement frequencies to calculate Neis genetic distance measures for all pairwise population comparisons and I constructed a neighbor-joining phenetic tree for the populations using these distances. Genetic distance between populations of A. sp. varvestris increases with geographic separation, and tree structure indicates that restricted gene flow only occurs in a downstream direction. The effect of unidirectional (downstream)gene flow on population genetic structure in riverine midges is described here for the first time. A genetic break is apparent between populations associated with 12 000-year-ol sediments originating in different ancient lakes. The geological break between lakes also coincides with a break in the species composition of the Axarus populations