Genetic enrichment of cardiomyocytes derived from mouse embryonic stem cells

Pluripotent embryonic stem cells (ESC) have the ability to differentiate into a variety of cell lineages in vitro, including cardiomyocytes. Successful applications of ESC-derived cardiomyocytes in cell therapy and tissue engineering were limited by difficulties in selecting the desired cells from the heterogeneous cell population. We describe a simple method to generate relatively pure cardiomyocytes from mouse ESCs. A construct comprising mouse cardiac α-myosin heavy chain (MHC) promoter driving the neomycin resistance gene and SV40 promoter driving the hygromycin resistant gene designated pMHCneo/ SV40-hygro, was stably transfected into mouse ESCs. The transgenic ESC line, designated MN6 retained the undifferentiated state and the potential of cardiogenic differentiation. After G418 selection, more than 99% of cells expressed α-sarcomeric actin. Immunocytological and ultrastructural analysis demonstrated that, the selected cardiomyocytes were highly differentiated. Our results represent a simple genetic manipulation used to product essentially pure cardiomyocytes from differentiating ESCs. It may facilitate the development of cell therapy in heart diseases.Key words: Embryonic stem cells, α-myosin heavy chain promoter, cardiomyocytes, differentiation, genetic enrichment

Malondialdehyde level and some enzymatic activities in subclinical mastitis milk

The purpose of this study was to evaluate the changes occurring in milk malondialdehyde (MDA) level and some enzymatic activities as a result of subclinical mastitis (SCM) in dairy cows. A total of 124 milk samples were collected from 124 lactating cows from the same herd in the period between the 2nd week after calving and the 10th week postpartum. They were classified by bacterial culture and the California mastitis test (CMT) as positive were deemed to have glands with SCM, and the periodic incidence rate of SCM was 26.6%. The most common bacterial isolates from SCM cases were Staphylococcus aureus (47%) and coagulase negative Staphylococci (CNS) (27%). The mean level of MDA and activities of lactate dehydrogenase (LDH) and alkaline phosphatase (ALP) were significantly higher in SCM milk than in normal milk, while the mean activity of glutathione peroxidase (GPx) was significantly lower in SCM milk than in normal milk. There were no differences in the activities of superoxide dismutase (SOD) and aspartate aminotransferase (AST) between normal milk and SCM milk. Therefore, the measurement of milk MDA level and GPx, LDH and ALP activities, appears to be a suitable diagnostic method for identifying SCM in dairy cows.Key words: Subclinical mastitis, mastitis diagnostic, etiology, malonaldehyde (MDA), enzym

Field-testing of the revised, draft South African Paediatric Food-Based Dietary Guidelines amongst mothers/caregivers of children aged 0–12 months in the Breede Valley sub-district, Western Cape province, South Africa

Objectives: To assess the appropriateness and understanding of the revised, draft South African Paediatric Food-Based Dietary Guidelines (SA-PFBDGs) amongst mothers/caregivers of children aged 0–12 months. Exposure to guidelines with similar messages, barriers and enablers to following of the guidelines were also assessed. Design: Qualitative data were collected from 14 focus-group discussions (FGDs), conducted in isiXhosa (n = 5), English (n = 4) and Afrikaans (n = 5), totalling 73 mother/caregiver participants. Setting: Worcester, Breede Valley sub-district, Western Cape province. Subjects: The study population included mothers/caregivers who were older than 18 years. Results: The majority of participants had previous exposure to variations of messages similar to the revised, draft SA-PFBDGs. Health platforms and practitioners (community health centres, antenatal classes, nurses, doctors) and social networks and platforms (family, magazines, radio) were mentioned as primary sources of information. Barriers to following the messages included: inconsistent messages (mainly communicated by healthcare workers), contrasting beliefs and cultural/family practices, limited physical and financial access to resources, poor social support structures and the psycho-social and physical demands of raising a child. Conclusion: The revised, draft SA-PFBDGs for the age range 0–12 months have been field-tested in English, Afrikaans and isiXhosa. The messages in some of the revised, draft SA-PFBDGs were not understood by the participants, indicating that a degree of rewording should be considered to facilitate understanding of the guidelines by the public. The National Department of Health should consider the findings of this study, and use these standardised message/s to optimise infant and young child feeding

Preparation of meso-tetra(4-benzenesulfonylphenyl) - porphyrin by the method of phase transfer catalysis

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Two rapid assays for screening of patulin biodegradation

Artículo sobre distintos ensayos para comprobar la biodegradación de la patulinaThe mycotoxin patulin is produced by the blue mould pathogen Penicillium expansum in rotting apples during postharvest storage. Patulin is toxic to a wide range of organisms, including humans, animals, fungi and bacteria. Wash water from apple packing and processing houses often harbours patulin and fungal spores, which can contaminate the environment. Ubiquitous epiphytic yeasts, such as Rhodosporidium kratochvilovae strain LS11 which is a biocontrol agent of P. expansum in apples, have the capacity to resist the toxicity of patulin and to biodegrade it. Two non-toxic products are formed. One is desoxypatulinic acid. The aim of the work was to develop rapid, high-throughput bioassays for monitoring patulin degradation in multiple samples. Escherichia coli was highly sensitive to patulin, but insensitive to desoxypatulinic acid. This was utilized to develop a detection test for patulin, replacing time-consuming thin layer chromatography or high-performance liquid chromatography. Two assays for patulin degradation were developed, one in liquid medium and the other in semi-solid medium. Both assays allow the contemporary screening of a large number of samples. The liquid medium assay utilizes 96-well microtiter plates and was optimized for using a minimum of patulin. The semisolid medium assay has the added advantage of slowing down the biodegradation, which allows the study and isolation of transient degradation products. The two assays are complementary and have several areas of utilization, from screening a bank of microorganisms for biodegradation ability to the study of biodegradation pathways

Association between Low Density Lipoprotein Receptor-Related Protein 2 Gene Polymorphisms and Bone Mineral Density Variation in Chinese Population

Low density lipoprotein receptor-related protein 2 gene (LRP2) is located next to the genomic region showing suggestive linkage with both hip and wrist bone mineral density (BMD) phenotypes. LRP2 knockout mice showed severe vitamin D deficiency and bone disease, indicating the involvement of LRP2 in the preservation of vitamin D metabolites and delivery of the precursor to the kidney for the generation of 1α,25(OH)2D3. In order to investigate the contribution of LRP2 gene polymorphisms to the variation of BMD in Chinese population, a total of 330 Chinese female-offspring nuclear families with 1088 individuals and 400 Chinese male-offspring nuclear families with 1215 individuals were genotyped at six tagSNPs of the LRP2 gene (rs2389557, rs2544381, rs7600336, rs10210408, rs2075252 and rs4667591). BMD values at the lumbar spine 1–4 (L1-4) and hip sites were measured by DXA. The association between LRP2 polymorphisms and BMD phenotypes was assessed by quantitative transmission disequilibrium tests (QTDTs) in female- and male-offspring nuclear families separately. In the female-offspring nuclear families, rs2075252 and haplotype GA of rs4667591 and rs2075252 were identified in the nominally significant total association with peak BMD at L1-4; however, no significant within-family association was found between peak BMD at the L1-4 and hip sites and six tagSNPs or haplotypes. In male-offspring nuclear families, neither the six tagSNPs nor the haplotypes was in total association or within-family association with the peak BMD variation at the L1-4 and hip sites by QTDT analysis. Our findings suggested that the polymorphisms of LRP2 gene is not a major factor that contributes to the peak BMD variation in Chinese population

Spatio-Temporal Interpolation Is Accomplished by Binocular Form and Motion Mechanisms

Spatio-temporal interpolation describes the ability of the visual system to perceive shapes as whole figures (Gestalts), even if they are moving behind narrow apertures, so that only thin slices of them meet the eye at any given point in time. The interpolation process requires registration of the form slices, as well as perception of the shape's global motion, in order to reassemble the slices in the correct order. The commonly proposed mechanism is a spatio-temporal motion detector with a receptive field, for which spatial distance and temporal delays are interchangeable, and which has generally been regarded as monocular. Here we investigate separately the nature of the motion and the form detection involved in spatio-temporal interpolation, using dichoptic masking and interocular presentation tasks. The results clearly demonstrate that the associated mechanisms for both motion and form are binocular rather than monocular. Hence, we question the traditional view according to which spatio-temporal interpolation is achieved by monocular first-order motion-energy detectors in favour of models featuring binocular motion and form detection

Is PTEN loss associated with clinical outcome measures in human prostate cancer?

Inactivating PTEN mutations are commonly found in prostate cancer, resulting in an increased activation of Akt. In this study, we investigate the role of PTEN deletion and protein expression in the development of hormone-refractory prostate cancer using matched hormone-sensitive and hormone-refractory tumours. Fluorescent in situ hybridisation and immunohistochemistry was carried out to investigate PTEN gene deletion and PTEN protein expression in the transition from hormone-sensitive to hormone-refractory prostate cancer utilising 68 matched hormone sensitive and hormone-refractory tumour pairs (one before and one after hormone relapse). Heterogeneous PTEN gene deletion was observed in 23% of hormone sensitive tumours. This increased significantly to 52% in hormone-refractory tumours (P=0.044). PTEN protein expression was observed in the membrane, cytoplasm and the nucleus. In hormone sensitive tumours, low levels of cytoplasmic PTEN was independently associated with shorter time to relapse compared to high levels of PTEN (P=0.028, hazard ratio 0.51 (95%CI 0.27–0.93). Loss of PTEN expression in the nucleus of hormone sensitive tumours was independently associated with disease-specific survival (P=0.031, hazard ratio 0.52, 95%CI 0.29–0.95). The results from this study demonstrate a role for both cytoplasmic and nuclear PTEN in progression of prostate cancer to the hormone-refractory state

Emodin Suppresses Migration and Invasion through the Modulation of CXCR4 Expression in an Orthotopic Model of Human Hepatocellular Carcinoma

10.1371/journal.pone.0057015PLoS ONE83