Semmelweis’ present day follow-up: Updating bacterial sampling and enrichment in clinical hygiene

Potentially dangerous antibiotic resistant contaminants have permanently penetrated at least well-off western populations. The danger is so evident that some hospitals have started to refuse accepting patients who carry such bacteria. Sampling and enrichment measures in hygiene monitoring must be updated as they are corner stones in handling the problems and safeguarding the health care units. Their patients, when exposed to microorganisms are strenuous to treat. Sometimes even this fails, if the infections are spreading in weakened patients. The present review summarizes currently used technologies and the abilities of bacteria to avoid detection. Improved protocols on environmental monitoring in healthcare units are required. They should be comparable with contamination control in industries. Actually these measures in health care should be even stricter because human lives are directly endangered as the resistance of especially elderly patients is low.     

Enrichment of Oct3/4‐positive cells from a human bronchial epithelial cell line

Most adult stem cells are in the G0 phase of the cell cycle, accounting for only a small percentage of the cells in the tissue. Thus, isolation of stem cells from tissues for further study represents a major challenge. The anti‐tumor drug 5‐fluorouracil (5‐FU) selectively kills proliferating cells, sparing cells in the G0 phase. Thus, the objective of this study was to determine whether 5‐FU can be used to enrich stem cells in a human bronchial epithelial (HBE) cell population in vitro. Side population (SP) cells were isolated from untreated HBE cells or HBE cells treated with 5‐FU, and the resulting cells were subjected to colony formation assays, culturing of cell spheres, and tumorigenicity assays. Expression of Oct3/4, Sox2, PCK, and β‐catenin were examined by Western blot analysis and immunofluorescence. Treatment with 5‐FU increased the percentage of SP cells from 0.3% to 1.5%, and the clonogenic ability of 5‐FU‐treated cells was more than twofold higher than that of HBE cells. Cells that survived after 5‐FU treatment exhibited a higher capacity for sphere formation. Furthermore, spheres formed from 5‐FU‐treated cells possessed the capacity to generate differentiated progenies. Cells treated with 5‐FU also exhibited tumorigenic potential, based on tumor formation assays in nude mice, and Oct3/4‐positive cell aggregates were identified in the resulting tumors. In this study, we have shown that 5‐FU treatment enriched the population of cells expressing the putative embryonic markers Oct3/4 and Sox2 and exhibiting nuclear accumulation of β‐catenin. Furthermore, 5‐FU‐treated cells expressed low levels of the epithelial differentiation marker PCK. Analysis of epigenetic modifications suggested that Oct3/4‐positive cells possessed characteristics of stem cells. These results demonstrate that treatment with 5‐FU can enrich the stem cell population present in a human bronchial epithelial cell line, and implicate combined treatment with 5‐FU and serum‐free medium as a new method for isolation of stem‐like cells from the HBE cell line.     

循环肿瘤细胞在前列腺癌中的新检测方法及其临床应用的研究进展

循环肿瘤细胞(CTCs)的检测不仅有助于肿瘤发生的早期预警,监测肿瘤的转移或复发,指导并判断肿瘤治疗效果,推断预后,而且可进一步研究肿瘤转移的机制。在肿瘤的临床治疗和转移机制的研究方面有重大的意义。目前检测血液中CTCs的传统方法有多种,但大多不能兼顾检测精确性、高效性、实用性和经济性。随着富集检测技术研究的进展,出现了一些新的检测方法,为临床肿瘤的治疗和机制研究提供了良好的平台。该文主要介绍几种新的CTCs的富集、检测和分析方法及其在前列腺癌患者中的临床应用。     

Systematic review and Meta-analysis of treating open angle glaucoma with gonioscopy-assisted transluminal trabeculotomy

AIM: To describe the complex, overlapping phenotype of four Chinese patients with inherited retinal dystrophies (IRDs) who harbored two pathogenic genes simultaneously. METHODS: This retrospective study included 4 patients affected with IRDs. Medical and ophthalmic histories were obtained, and clinical examinations were performed. A specific Hereditary Eye Disease Enrichment Panel (HEDEP) based on exome capture technology was used for genetic screening. RESULTS: Four patients were identified to harbor disease-causing variants in two different genes. Patient RP01-II:1 exhibited both classical ABCA4-induced STGD1 and USH2A-associated RP, patient RP02-III:2 exhibited both classical ABCA4-induced STGD1 and CDH23-associated RP, patient RP03-II:1 exhibited both USH2A-induced arRP syndrome and SNRNP200-induced adRP, and patient RP04- II:2 exhibited USH2A-induced arRP syndrome and EYS-induced arRP at the same time. CONCLUSION: Our study demonstrates that genotype–phenotype correlations and comprehensive genetic screening is crucial for diagnosing IRDs and helping family planning for patients suffering from the disease.     

Discovery of Cellular RhoA Functions by the Integrated Application of Gene Set Enrichment Analysis

The small GTPase RhoA has been studied extensively for its role in actin dynamics. In this study, multiple bioinformatics tools were applied cooperatively to the microarray dataset {"type":"entrez-geo","attrs":{"text":"GSE64714","term_id":"64714"}}GSE64714 to explore previously unidentified functions of RhoA. Comparative gene expression analysis revealed 545 differentially expressed genes in RhoA-null cells versus controls. Gene set enrichment analysis (GSEA) was conducted with three gene set collections: (1) the hallmark, (2) the Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway, and (3) the Gene Ontology Biological Process. GSEA results showed that RhoA is related strongly to diverse pathways: cell cycle/growth, DNA repair, metabolism, keratinization, response to fungus, and vesicular transport. These functions were verified by heatmap analysis, KEGG pathway diagramming, and direct acyclic graphing. The use of multiple gene set collections restricted the leakage of information extracted. However, gene sets from individual collections are heterogenous in gene element composition, number, and the contextual meaning embraced in names. Indeed, there was a limit to deriving functions with high accuracy and reliability simply from gene set names. The comparison of multiple gene set collections showed that although the gene sets had similar names, the gene elements were extremely heterogeneous. Thus, the type of collection chosen and the analytical context influence the interpretation of GSEA results. Nonetheless, the analyses of multiple collections made it possible to derive robust and consistent function identifications. This study confirmed several well-described roles of RhoA and revealed less explored functions, suggesting future research directions.     

On chip cell separator using magnetic bead-based enrichment and depletion of various surface markers

This paper presents an on-chip magnetic cell sorting system for the sorting of cells based on a variety of surface markers. A polymer lab on a chip integrated with an electroplated array of Ni/Fe permalloy has been designed, fabricated, and characterized for the separation of cell substitutes at a variety of flow rates and incubation times. The system sequentially labels cell substitutes with magnetic beads and sorts them, repeating this process to sort for a variety of surface markers. Flow rates and incubation times were varied to characterize the system and produce the best combination of high specific capture and low nonspecific capture. The separation system developed on polymer is selective and efficient while being low cost, portable, and fabricated in a modular structure that can be integrated with other cell handling processes.     

Opioid addiction: Who are your real friends?

Opioid addiction is a chronic and relapsing mental health disorder. However, only some individuals exposed to opioids, either recreationally or during the course of pain management, will develop addiction. The reasons why some individuals develop addiction and some are spared are not fully understood. Studies indicate that it is likely a combination of genetic predispositions and environmental conditions. Given the role of environmental factors in human addiction, this review examines the role of social environments and social interactions in the development of opioid addictive-like behaviors in rodent studies. To date, three major behavioral approaches have been used in these studies, namely social isolation, environmental enrichment, and social housing with a variety of cage-mates that differ in their drug administration conditions. This review highlights the importance of an individual’s social network in influencing the outcomes of drug abuse and the need to further elucidate the molecular mechanisms underlying these effects. Better understanding is likely to contribute to the development of novel and more effective treatments for addiction disorders.     

Aspects of the biomonitoring studies using mosses and lichens as indicators of metal pollution

It has been shown that mosses and lichens, in spite of all disadvantages, are good tools for air pollution monitoring, but best results could be achieved while using both of them together, because of differences in their metal uptake and retention. The researcher dealing with biomonitoring faces many difficulties; for example, in the use of lichens, those of similar composition are not easy to find, because of the differences caused by the tree on which the lichens are growing. Also, terrestrial moss is not always a good choice because of variations in its composition caused by the area it is growing on. The "bag" technique seems to be suitable in many cases because it gives the possibility of comparing wet and dry deposition and there are no differences in its content at the beginning of research. The bag construction should be made to keep the humidity of the sample stable to avoid its drying. For a complete and informative data set the biomonitor content should be compared with precipitation, particulate matter fractionation, and speciation data and also with medical statistics to evaluate the correlation between the amount of pollutants in the atmosphere and the human reaction to them.     

Specific magnetic bead-based capture of free fetal DNA from maternal plasma

An automated magnetic capture hybridization (MCH) method for the extraction and enrichment of fetal RHD specific DNA fragments from maternal plasma was developed using plasma from 1000 D-negative pregnant women. A real time PCR protocol for RHD exon 7 was applied. MCH was compared with the QIAamp DSP Virus Kit (QIAamp) as a reference. Compared with the QIAamp method, the percentage of fetal DNA increased from 2.86% to 4.83% (p < 0.05, n = 8). The 95% detection limit of MCH was determined at 286 pg/ml (43 geg/ml) compared with 138 pg/ml (21 geq/ml) for the QIAamp DSP Virus Kit.