With this goal in mind, we will examine evidence from other systems that present cell growth. Surface tension driven cellular patterns (by short, ‘‘foam-like’’ systems) have been studied in the current literature associated with grain growth in polycrystalline materials and cellular development in foams. Although there is not complete agreement in the way these complex systems evolve, recently, a theoretical treatment for two-dimensional grain growth in a stochastic framework has been proposed . Also, there are other proposed theoretical models . An appropriate vision of the ‘‘state of the art’’ on this matter appears in some seminal reviews [10-11]. On the other side, the Physics of foams have been comprehensively revised in a recent book  and reviewed in current works appearing in the literature [13,14]. As well, there have been approaches that simulate the grain growth behavior in polycrystalline materials by using soap froth patterns [15,16].
indicating that 4R tau does not simply replace the function of 3R tau. The functional difference between these isoforms is not clear. To understand the pathway linking 4R tau over-expression to neurodegeneration, there is a need to identify an appropriate model where the pattern of tau isoform expression is comparable to that of adulthuman brain. The absence of a consistent amount of 3R tau in the adult rodent brain makes it unsuitable to address questions related to the imbalance between 3R and 4R tau. Inhuman neuroblastoma cell lines such as SH SY5Y, it is difficult to obtain an adult tau isoform pattern; even after several weeks of differentiation, the shortest 3R and 4R isoforms are the predominant species. Furthermore, transgenic mice expressing the entire human tau gene also do not show the tau isoform expression pattern found inadulthuman brain [12,13]. Against this background, hESC-derived neurons could be an alternative in vitro model to study tau physiology and pathology. We have studied the temporal distribution and expression of 3R and 4R tau
Neurotransmitter Identification and Axonal Growth The predominant neurotransmitter phenotype of differ- entiated NSCs is GABAergic, although a sizeable minority of these cells acquires cholinergic phenotypes. It has been reported that ‘‘priming’’ of human NSCs prior to grafting may advance them more fully to cholinergic fates postgraft- ing [35,36], but the likely advanced maturation state of these cells pregrafting and unknown survival rates and grafting efﬁciency does not allow direct comparisons with the results of this study. GABAergic and cholinergic neurotransmitter signatures appear to belong to neurons with different cytologies, i.e., GABAergic neurons have smaller bipolar cytologies whereas cholinergic neurons are larger and multi- polar. However, all shapes and sizes between these two types have been encountered and, in a few cases of grafts growing outside the spinal cord parenchyma, we have seen a continuous pattern of differentiation in colonies in which GABAergic neurons predominate in the periphery and cholinergic neurons at the center. Therefore, it is unlikely that these two phenotypes belong at the end points of distinct differentiation lineages and it is possible that some GABAer- gic neurons could turn into cholinergic nerve cells given the proper instructive signals. Another argument against the idea of the GABAergic phenotype as an end-point fate is the fact that a majority of HNu (þ) cells in the spinal cord extend long axons, whereas GABAergic spinal cord neurons in mature animals are interneurons [37,38].
A decrease in functional beta-cell mass is a key feature of type 2 diabetes. Glucagon-like peptide 1 (GLP-1) analogues induce proliferation of rodent beta-cells. However, the proliferative capacity of human beta-cells and its modulation by GLP-1 analogues remain to be fully investigated. We therefore sought to quantify adulthuman beta-cell proliferation in vitro and whether this is affected by the GLP-1 analogue liraglutide. Human islets from 7 adult cadaveric organ donors were dispersed into single cells. Beta-cells were purified by FACS. Non-sorted cells and the beta-cell enriched (‘‘beta-cells’’) population were plated on extracellular matrix from rat (804G) and human bladder carcinoma cells (HTB9) or bovine corneal endothelial ECM (BCEC). Cells were maintained in culture+/2liraglutide for 4 days in the presence of BrdU. Rare human beta-cell proliferation could be observed either in the purified beta-cell population (0.05160.020%; 22 beta-cells proliferating out of 84’283 beta-cells counted) or in the non-sorted cell population (0.05560.011%; 104 proliferating beta- cells out of 232’826 beta-cells counted), independently of the matrix or the culture conditions. Liraglutide increased human beta-cell proliferation on BCEC in the non-sorted cell population (0.08260.034% proliferating beta-cells vs. 0.01760.008% in control, p,0.05). These results indicate that adulthuman beta-cell proliferation can occur in vitro but remains an extremely rare event with these donors and particular culture conditions. Liraglutide increases beta-cell proliferation only in the non- sorted cell population and only on BCEC. However, it cannot be excluded that human beta-cells may proliferate to a greater extent in situ in response to natural stimuli.
The Statistical Package for Social Scientists (SPSS) software (version 15) was used in testing whether or not the means of dependent variables were significantly different among groups. The total % yield of nitrogen, phosphorus and potassium of the stored urine over the 6-month study period were analysed. This was indicative of when the urine could be used for crops that require proportionally high percentage of nitrogen, phosphorus or potassium. The significant difference in yield of NPK between male and female urines was also established for each month over the 6 months study period. If the overall ANOVA was significant and a factor had more than two levels, a post-hoc multiple comparisons follow up test was carried out using Least Significance Difference (LSD) or Duncan’s Multiple Range Test (DMRT). In all cases, significance was determined at the 95% confidence level. One-way analysis of variance was performed to assess the differences among means, with a significance level of 5% (p< 0.05).
In humans, NANOS3 is expressed in fetal and adult ovaries during multiple stages of oogenesis and in vitro stud- ies of human embryonic stem cell-derived germ cells have shown coexpression of NANOS3 in nuclei with important germ cell factors such as BLIMP1, VASA, and STELLA in the cytoplasm . In these cells, reduction of NANOS3 expression resulted in altered gene expression patterns and reduced number of cells in active division, suggesting that in humans too NANOS3 has an important function in germ cell maintenance and survival. Recently, a heterozygous p.Arg153Trp NANOS3 mutation was reported in a 23-year- old Chinese woman out of a cohort of 100 women with POI . Familial segregation was not deined, but the mutation was not found in 200 ethnically-matched controls. In vitro studies suggested decreased protein stability due to the p.Arg153 >Trp mutation, leading the authors to postulate a mechanism of reduced dosage of NANOS3 expression in the ovaries leading to decreased PGC population and resulting in POI .
Drug release from DTX-BIO-BSA-NPs was studied and compared with drug release/diffusion from DTX suspension using a dialysis bag method (Liu et al., 2011). The phosphate-buffered saline (0.1 M, pH 7.4) at 37 ºC was used to determine the docetaxel release from the nanoparticles. After adding DTX-BIO-BSA- NPs suspension (0.5 mg/mL, 2 mL, without trehalose) or docetaxel solution (0.05 mg/mL, 2 mL) into Slide- A-Lyzer dialysis cassettes (Sigma-Aldrich, St. Louis, USA), the dialysis cassettes were then immersed in a 250 mL beaker containing 200 mL release buffer, which was placed in an incubator shaker (Jiangsu Jintan Medical Instruments Co., Ltd., Jintan, China) at 37 ºC and 120 rpm. Later, with drawn from the beaker, 50 mL release buffer was replaced with 50 mL fresh release buffer by regular intervals. Each collected release buffer sample was concentrated to 1 mL by rotary evaporator, docetaxel was resolved by high-performance liquid chromatography and extracted from the removed medium after dividing into the organic phase. After being added to the medium, 5 mL chloroform was fully vortexed for 5 min and allowed to stand for 15 min for phase separation. Upon phase separation, the denser organic chloroform layer was carefully partitioned from the aqueous buffer phase and permitted to evaporate at the indoor temperature overnight. The dried sample containing docetaxel was then dissolved in 4 mL of methanol and analyzed by high-performance liquid chromatography. It was plotted to a profile showing cumulative drug release on the basis of time. And each diffusion experiment was operated repeatedly in triplicate, the average values and normal deviations were calculated in detail.
tests yielded the following results: lactate dehydrogenase, 215U/l; beta-2-microglobulin, 5.52 mg/dl; calcium, 14.4 mg/dl; phosphorus, 5.2 mg/dl; uric acid, 11.1 mg/dl; and albumin, 2.9 g/dl. The presence of anti-HTLV-I antibodies was detected by means of ELISA (Ortho-Clinical Diagnostics, Raritan, New Jersey, USA) and was conirmed by Western blot (Genelabs Diagnostics, St. Ingbert, Germany). Myelogram and bone marrow biopsy showed no evidence of any neoplasm. Skeletal X-ray revealed multiple lytic lesions in the skull (Figure 3) and right femur. Digestive endoscopy showed esophagitis caused by monilia. Abdominal tomography revealed retroperitoneal lymph nodes and a biopsy obtained from the right axillary lymph node was compatible with T-cell lymphoma. Immunohistochemistry identiied the presence of CD3 + lymphocytes.
MSC culture in adipogenic differentia- tion medium led to the appearance, after 7 days, of larger rounded cells presenting nu- merous fat vacuoles in the cytoplasm visual- ized by Sudan III staining. The number of these cells increased continuously up to the 20th day of culture and remained stable for more than two months of culture (Figure 2). The osteogenic stimulus of MSC led to the appearance, after 15 days of culture, of refringent crystals on the cells, better visual- ized by silver nitrate staining. Staining with alkaline phosphatase and silver nitrate per- mitted us to demonstrate the presence of osteocytic differentiation in the induced MSC culture (Figure 2).
The ability to generate TDECs under defined in vitro conditions answers several questions that cannot be readily addressed using in vivo models where the tumor microenvironment is often heterogenous, subject to rapid change [1,33] and where TDEC formation is likely to compete with additional vascular remodeling processes. These questions include the nature and interrelationships of the inductive signals for TDECs and their timing. Obvious candidates for such signaling molecules include EC-specific growth factors such as VEGF, bFGF and TGF-beta, as well as hypoxia, all of which are strong inducers of tumor neovascularization [3-5,11-14]. Our results indicate that, at least in vitro, neither the growth factors supplied by EC-specific growth medium nor hypoxia alone are particularly potent inducers of the tumor cell to TDEC transition but that, in combination, they are highly synergistic. This is not entirely unexpected as these factors have been long known to be necessary for the generation and maintenance of the tumor Figure 3. Hypoxia, EC-specific growth medium, and nutrient deprivation induce Tie2-dependent EGFP expression in tumor cells. Separate cultures of H460 and OVCAR3 tumor cells were stably co-transfected with Tie2-EGFP plasmid and pFR400 encoding a mutant form of dihydrofolate reductase [36,37] and selected in G-418 and increasing concentrations of methotrexate to allow amplification of the two tandemly integrated vectors and a corresponding increase EGFP signal intensity. Cells were exposed to conditions shown previously to induce the maximal TDEC phenotype (i.e., condition 3 for H460’s and condition 4 for OVCAR3) and compared to control cells grown under standard conditions. (A) Fluorescence micrographs of each cell line after five days of growth under each set of conditions. (B) Flow cytometric analysis of the same cells. Representative results of at least three independent experiments are depicted. Scale bar = 25 um.
Pattern recognition tries to extract information from ecological patterns and is the first step for inferring ecological processes from patterns (e.g., Boyero et al., 2015).Since the scope of ecological pattern is extensively broad, there is no universal methodology in describing these patterns. In general, depicting ecological patterns using a simple metric or statistical distribution is a preferred way, such as the branching diagram used in phylogenetic analysis, species abundance distribution in a community, spatial-correlation of species distributions (Hui, 2009; Gao, 2014), network architectures of complex ecosystems (Zhang et al., 2011; Minoarivelo et al., 2014; Nuwagaba et al., 2015), and various biodiversity indictors. Unfortunately, the relationship between ecological pattern and process is neither one-to-one nor linear (e.g. Hui and McGeoch, 2014): a single ecological process can produce different context-dependent ecological patterns, and similar ecological patterns can also be driven by different ecological processes (Brown et al., 2011).
(Universidade Federal de Pelotas, Brazil)*; Giovanni Viegi (Italian National Research Council, Italy)*; Lucie Viet (National Institute for Public Health and the Environment, The Netherlands)*; Eira Viikari- Juntura (Finnish Institute of Occupational Health, Finland)*; Paolo Vineis (Imperial College London, UK)*; Jesus Vioque (Universidad Miguel Hernandez, Spain)*; Jyrki K Virtanen (University of Eastern Finland, Finland)*; Sophie Visvikis-Siest (INSERM, France)*; Bharathi Viswanathan (Ministry of Health, Seychelles)*; Peter Vollenweider (Lausanne University Hospital, Switzerland)*; Sari Voutilainen (Uni- versity of Eastern Finland, Finland)*; Ana Vrdoljak (UHC Zagreb, Croatia)*; Martine Vrijheid (ISGlobal Centre for Research in Environmental Epidemiology, Spain)*; Alisha N Wade (University of the Witwatersrand, South Africa)*; Aline Wagner (University of Strasbourg, France)*; Janette Walton (University College Cork, Ireland)*; Wan Nazaimoon Wan Mohamud (Institute for Medical Research, Malaysia)*; Ming-Dong Wang (Public Health Agency of Canada, Canada)*; Qian Wang (Xinjiang Medical University, China)*; Ya Xing Wang (Beijing Tongren Hospital, China)*; S Goya Wannamethee (University College London, UK)*; Nicholas Wareham (University of Cambridge, UK)*; Deepa Weera- sekera (Ministry of Health, New Zealand)*; Peter H Whincup (St George’s, University of London, UK) *; Kurt Widhalm (Medical University of Vienna, Austria)*; Indah S Widyahening (Universitas Indonesia, Indonesia)*; Andrzej Wiecek (Medical University of Silesia, Poland)*; Alet H Wijga (National Institute for Public Health and the Environment, The Netherlands)*; Rainford J Wilks (The University of the West Indies, Jamaica)*; Johann Willeit (Medical University Innsbruck, Austria)*; Tom Wilsgaard (UiT The Arctic University of Norway, Norway)*; Bogdan Wojtyniak (National Institute of Public Health- National Institute of Hygiene, Poland)*; Jyh Eiin Wong (Universiti Kebangsaan Malaysia, Malaysia)*; Tien Yin Wong
Abstract Three patients (males, black, ages 37, 40 and 57) attended a university clinic with a progressive paraparesis of obscure origin. One patient who referred disease duration of more than 16 years, showed diminished deep reflexes, bilateral Babinski's sign, diminished sensation of vibration, abnormal bladder function and back pain. The other two patients (with one and six years of disease duration) complained of weakness in one leg, increased deep reflexes and back pain. Babinski's sign and bladder disturbance were also present in the patient with six years of disease. Blood samples tested by an enzyme immune assay and a discriminatory Western blot were positive for HTLV-I. The familial analysis of one patient showed a possible pattern of sexual and vertical transmission of the virus. To the best of our knowledge, these are the first cases of a proven association between HTLV-I and TSP/HAM in Belem, Para, and emphasize the need to actively look for cases of neurological disease associated to the virus.
During fetal development, the human liver is exceptionally erythropoietic [3,4], which wanes early in life. We observed only low levels of erythropoiesis in the adult chimeric liver. However, previous reports have noted depressed human erythropoiesis in chimeric mice , and our own experience analyzing the BM of chimeric NSG mice confirms a trend toward lower levels of erythropoiesis than either myelopoiesis or B-lymphopoiesis (Fig. 7 and unpublished data). There is also controversy as to whether murine erythropoietin is as active on human as mouse cells [57- 59]. Any possible deficiency in EPO stimulation may also be compounded by the lack of cross-reactivity of murine GM-CSF and other growth regulatory molecules leading to depressed human erythropoiesis in the murine BM and liver. Moreover, oncostatin M is thought to play a major role in the inhibition of post-natal erythropoiesis and hepatic hematopoiesis in general .
dexamethasone and TGF-b1. Thus, TGF-b3 appears to be superior to TGF-b1 (25). Our decision to use TGF-b3 was also based on a previous study from our group in which TGF-b3 was more efficient for inducing differentiation of MSCs derived from UCB into chondrocytes (9) than other growth factors (TGF-b3 was combined with dexamethasone to induce chondrogenesis). As a result, we were able to successfully induce the expression of type II collagen, aggrecan and SOX-9. In this study, chondrogenesis was induced in HAF-derived MSCs stimulated with TGF-b3 over 21 days using the micro- mass system. We confirmed this differentiation by analyzing the expression of the most important genes for the formation of articular cartilage (SOX-9, collagen type II and aggrecan) and confirmed the production of collagen type II at the protein level using histology or western blotting. Compared with normal human cartilage (used as a standard), cells under the micromass system showed significantly higher expression of the SOX-9 gene, as depicted in Figure 3A. This result indicates that it is possible to reproduce human chondrogen- esis in vitro, as SOX-9 is a key factor in inducing this process. Our results are reinforced by another study (5) in which the authors detected SOX-9 expression in cells under the micro- mass system but did not find a significant difference compared with that in cells under the pellet system, thereby indicating that transcription factor SOX-9 is essential for inducing chondrogenesis in both high-density systems.
Anterior lamellar keratoplasty is indicated for corneal opacities of the anterior or middle stroma, and it can be superficial (at the level of the stroma-stroma interface, referred to as superficial anterior lamellar keratoplasty [SALK]) or deep (at the stroma-Descemet membrane interface, referred to as deep anterior lamellar keratoplasty [DALK]). It is indicated in diseases such as keratoconus, pellucid marginal degeneration, stromal dystrophies, and ectasia after refractive surgery with healthy posterior layers (5,6,7) . Even though the technique has a longer learning curve, it offers significant
c) Descriptive analysis of the variables: In this stage, due to the variables identiied in CCM to be evaluated by the three observers, the measurements were aggregated with the purpose of obtain- ing a diagnosis for each individual (consensus among observers). So for the number of ibers and LCs, the measures were grouped using the “average” of observers 1 and 3, with observer 2 exclud- ed due to low correlation with the others (see reliability analy- sis results). Categorical variables (tortuosity and thickness) were grouped using the measure of the “mode.” As the concordance rate was not 100% between observers, all were included with the third observer’s evaluation used in non-concordant cases. he variables of the study were explored and evaluated by age and sex. For continuous variables, mean comparisons were performed with the normality of those tested with the Kolmogorov-Smirnov and Shapiro tests. he diference of means was evaluated us- ing the Mann-Whitney U or Kruskal-Wallis H tests for variables with normal and non-parametric distributions, respectively. he relationships between age variables and CCM parameters were calculated by Pearson correlation (normal distributions) and Spearman (nonparametric variables). In relation to qualitative variables (categorical variables), the percentage diferences were evaluated using chi-square statistics. Fisher’s exact test was con- sidered when the expected frequencies were below 5%. he inlu- ences of age and sex on CCM parameters were also investigated using simple linear regression and multiple (continuous and nor- mally distributed variables) or generalized linear models (discrete or binary). Bivariate and multivariate analyzes were performed. he model it was performed using ANOVA (nested models) and the Akaike information criterion (AIC, not nested models). In all analyses, p < 0.05 was considered signiicant.
The implementation of prophylaxis with regular factor VIII (FVIII) replacement for patients with hemophilia radically changed the clinical presentation of this condition, from a dis- ease characterized by progressive disabling musculoskeletal complications, to one compatible with an active and virtually normal lifestyle. In Brazil, although most adults with severe hemophilia still suffer the impact of musculoskeletal com- plications in their quality of life, 1 the recent implementation
G232A mutation in the TRE1 element may enhance indirect binding to Tax via the CREB family of cellular transcription factors, thereby promoting the prolifera- tion of infected cells and resulting in an increase in patient PvLs. However, the validity of this hypothesis needs to be investigated in an independent series of experiments comparing the expression and activation of the Tax gene in HTLV-1-infected T cells with or with- out the G232A mutation. Previous reports have shown that mutations that abolish Tax effects are all localized in the CREB of the 21 bp repeats . Montagne et al. demonstrated that mutations introduced into domain A or C can severely impair the ability of the Tax protein to transactivate different promoters . Taken together, these data suggest that the TRE-1 element plays an important role in the activation of HTLV-1 gene expression.
As mentioned above, in contrast to the A549-NT cells, in addition to CEACAM1, about 15 percent of the A549-T cells also expressed CEACAM6 on their cell surface, and the alterations in contact inhibited growth pattern could be reversed in A549-T cells by shRNA mediated knock down of CEACAM6 expression. CEACAM6 expression has already been inversely correlated with cellular differentiation and has been identified as a marker for decreased survival in patients with CEACAM6 expressing cancers.[35,37,38] In a series of experiments, Duxbury and associates have demonstrated that CEACAM6 over-expression in pancreatic cancer is a determinant of cell proliferation and cell invasiveness. On the other hand, targeting CEACAM6 results in decreased tumor growth, and inhibits metastases in a mouse xenograft model. Our results are in accordance with these findings, but in addition we show for the first time that CEACAM6 acts as an inducer of proliferation in A549 cells, putatively by interfering directly or indirectly with the contact- inhibiting signal triggered by CEACAM1-4L. These data also support previous data implying that CEACAM6 over-expression inhibits the tissue architecture surveillance mechanism known as anoikis. In principle A549 cells were able to express GPI anchored CEACAMs as demonstrated by the generation of A549- CEACAM5 cell lines (Figure 5A). However, our finding that the survival of detached A549-CEACAM5 cells was significantly decreased was in contrast to the results published by Camacho- Leal and Stanners, who demonstrated that the GPI anchor of CEACAM5 mediates anoikis inhibition. This different result may be based on the fact that A549 cells represent a humancell