Health, and values obtained were scored on a scale of items according to the SomaticCell Score (SCS). Data were processed by Microsoft Excel software program from Microsoft Office Professional Plus (2010) and the statistical package STATGRAPHICS Plus 5.1. Values from SCC and SCS (985,0 ± 15,0 and 4,61 ± 0,03, respectively) evi- denced the presence of mastitis in an increasing annual trend at all levels (stabled and individual dairy cows, and milk tanks). SCC and SCS significant differences between state farms (1 026,3 ± 16,1 and 4,67 ± 0,03, respectively) and private farms (539,3 ± 26,7 and 3,82 ± 0,0, respectively) were detected. Losses in milk production were esti- mated in 15 % based on the average value for somaticcell count.
1992). The first offspring produced by the transfer of a cultured cell line was reported in 1996 (Campbell et al., 1996b). In this study, cells were derived from the embryonic disk of an in vivo produced day-9 sheep embryo. The cells, which remained in culture in vitro for a prolonged period of time (6 to 13 subpassages) adopted an epithelial morphology prior to being used as donors for the nuclear transfer procedure. The significance of this study was threefold: (1) it demonstrated that differentiated cells, cultured for prolonged periods of time had the ability to be reprogrammed and originate a new individual, (2) it proved cultured cells could be induced to enter and temporarily arrested at a so-called G0 or quiescent state and (3) it led the way for the production of Dolly, the first mammal cloned from an adult, differentiated somaticcell. In 1997, the world was fascinated by the birth of Dolly, a sheep created not from the fertilization of an oocyte and a sperm, but by the transfer of a nucleus from a fully differentiated somaticcell into a mature oocyte devoid of its own nuclear DNA by the process of NT (Wilmut et al., 1997). In their pathway to produce Dolly, Dr. Wilmut and colleagues learned to appreciate the cleverness of DNA being able to remodel and reprogram itself, given the appropriate conditions and timing. The announcement of the birth of a healthy animal cloned from a differentiated adult somaticcell ignited a firestorm of public and scientific interest in the field. This remarkable achievement led to an explosion of studies dealing with cell-cycle regulation, sources of and techniques of establishing stem cell lines and on early embryo development. Since Dolly, several other mammalian species have been cloned from differentiated somatic cells, including the cow (Cibelli et al., 1998; Kato et al., 1998), mouse
Introduction: The objective of the study was to evaluate the effect of exogenous β-mannanase addition on dry matter intake (DMI) in the pre- and postpartum periods, milk yield (MY) and milk composition, urea nitrogen content (NU) and somaticcell count (SCC) in Holstein-Friesian cows. Method: Thirty cows were used in the study (body weight (BW)=781±83 kg; of more than two lactations) of approximately 260 d of gestation. The cows were stratified by BW and their previous MY and randomly assigned to one of two treatments: 1) Total mixed ration (corn-silage of corn), Control; and 2) Control+0.10% of β-mannanase (CTCZYME; Seoul, Korea).
Design/methodology/approach: 48 samples of bulk milk from different dairy farms and cheese factories in Tabasco, Mexico were taken randomly. Measured parameters were: California Mastitis Test (CMT), the somaticcell count (CCS), fat, protein, non-fat solids, lactose. The values of CCS were grouped in 3 intervals: (500,000), (500,000-1,000,000), (1,000,000) to be compared with qualitative data of CMT (0, 1, 2) in a two-way contingency table. Milk samples were also grouped in 4 categories according to their number of somatic cells A (0-250,000), B (251,000-500,000), C (500,000- 750,000) y D (750,000) to compare fat, protein, non-fat solids, lactose by one-way ANOVA and Tukey test at a 5% confidence level.
This study demonstrates the use of nuclear somaticcell transfer to produce the first cloned cattle in Peru. Skin fibroblasts and cumulus cells from adult donors were obtained for use as carioplasts; likewise, oocytes obtained from ovaries in the slaughterhouse were matured in vitro for 24 h. The mature oocytes were incubated 2 h in demecolcin (2.5 µg/ml) to promote cone formation with the metaphase plate and to guide manual enucleation. The zona pellucida in pronase (2 mg/ml) was removed for 3 min. The enucleation was manual with a microblade dividing the ova into two halves, where the nucleus-lacking halves were fused by the «sandwich» method (cytoplast–fibroblast– cytoplast). The reconstructed structures were chemically activated by incubation for 5 min in 7% absolute ethanol, followed by 5 h of cytochalacin B (5 µg/ml) and cycloheximide (10 µg/ml). The structures were cultured for 7 d until the blastocyst incubation/hatching phase. Seven blastocysts were transferred to six synchronized recipient cows seven days after ovulation. The permanence of four and three embryonic vesicles was achieved until days 28 and 60, respectively. Two calves were born from embryos reconstructed with skin cells and cumulus cells. By the genotype analysis using 15 markers (SSR) for cattle, it was confirmed that cloned calves were derived from donor cell lines.
A total of 32 goats (11, 17 and 4 in 1st, 2nd and ≥ 3 lactations, respectively; 20 healthy and 12 with uni- lateral IMI caused by coagulase-negative staphylococci; all goats seronegative for caprine arthritis- encephalitis virus) were monitored daily, for 53 days, the somaticcell count (SCC) by gland and, weekly, the bacteriological analysis by gland. In healthy udders was considered there was a transient elevation of SCC (TESCC) when counts increased in both glands, at least 2.5 times that of previous days, reaching over 1 million cells/ml (0.7 million cells/ml in primiparous goats). To identify the TESCC in unilaterally infected udders was ascertained that the increase in SCC noted above occurred in the healthy gland, and counts also increased in the infected gland. Results confirmed the existence of non- infectious TESCC in goats. Although its duration was variable, usually it ranged between 1 and 3 days. Nearly 60% of goats showed a case of TESCC, appearing in both, healthy goats as in infected ones, as well as in primiparous and in multiparous goats.
The totipotent character of plant cells allow that any differentiated cells that retains its nucleus has the ability to regenerate an entire new plant by organogenesis or somatic embryogenesis (SE) (Reynolds 1997, Fortes & Pais 2000). SE is the developmental process by which bipolar structures that resemble zygotic embryos are developed from haploid or diploid somaticcell through an orderly embryological stage without gametes fusion (Jiménez 2001, Jiménez 2005, Quiroz-Figueroa et al. 2006, Namasivayam 2007). Two types of somatic embryogenesis are recognized: direct somatic embryogenesis (DSE) and indirect somatic embryogenesis (ISE). DSE is characterized by the induction of somatic embryos directly from pro-embryogenic cells from leaves, stem, microspores or protoplasts without the prolifer- ation of calli, whereas in ISE somatic embryos are developed from friable embryogenic calli (Jiménez 2001, Molina et al. 2002, Quiroz-
Assessing graft loss is a particularly challenging aspect of islet transplantation due to the many mechanisms eliciting islet injury and the lack of tools to measure beta cell mass in vivo 1 . We have described a 14% islet mass loss during culture associated with cell fragmentation and disintegration 29 . Another important islet loss event occurs immediately after infusion, largely due to cell hypoxia and inflammatory responses following transplantation. In particular, IBMIR accounts for significant graft attrition at this early stage and is reported to reach up to 70% of the initial preparation within the first 24hrs, when using the intraportal route
Mannosylation facilitates uptake and internalization of immunogenic peptides by antigen-processing cells expressing mannose receptors at their surface, such as DC-SIGN, a lectin that plays a key role in the immune response against different pathogens. This internalization, processing and subsequent MHC pre- sentation may result in a strong T cell stimulation. Here, we hypothesized that combining mannose glycodendrons with multivalent presentation of peptide epitopes in a likewise dendron format would yield hybrid constructs, named glycodendropeptides (GDPs), with the capacity to enhance peptide immunoge- nicity, hence providing a novel and versatile platform for applications in immunotherapy. Thus, GDPs of dif- ferent valencies displaying the NP 366–374 epitope, a conserved sequence from the influenza A virus nucleo-
NOTA: El IMMS-CELL usará el número de teléfono de “Datos” suministrado con el servicio. Normalmente, Las cuentas GSM proporcionan múltiples números de teléfono, aunque muchos clientes y comerciantes no son conscientes de ello. No configure el Sitio para usar Voz o Fax opcionales… sólo los números de Datos funcionarán correctamente.
(in fact, tumorigenic potential is a major drawback for their use in ther- apy). Naturally occuring pluripotent stem cells are derived from the inner cell mass of an embryo in the blastocyst stage and termed em- bryonic stem cells. As the derivation of these cells requires necessar- ily the destruction of a human embryo, they are the center of intense ethical debate. For the purposes of this article we will not discuss em- bryonic stem cells and we will focus our biobanking topic to other types of pluripotent stem cells and adult stem cells. Regular, terminally dif- ferentiated somatic stem cells (such as skin fibroblasts) can be induced to convert to a pluripotent state simply by the forced expression of a few transcription factors (Oct4, Sox2, Klf4 and c-Myc were the first de- fined, though there are a number of combinations and methods avail- able today) to generate the so called induced Pluripotent Stem Cells (iPSCs). iPSCs present the same characteristics of embryonic stem cells, without the ethical concerns raised by their derivation; further- more, they may be obtained from the same individual subject for ther- apy, preventing immunological rejection. Extended discussion of biobanking issues associated with the generation of iPSCs and biobank- ing will be provided later on.
Tumor cells are exposed to a continuum of oxygen concentrations and consequently solid tumors are comprised of three tissue regions: the normoxic, hypoxic and necrotic (Figure 1). Situated nearby functional blood vessels normoxic cells are typically viable and proliferative. At distances of 150 µm from patent blood vessels cells may become anoxic, giving rise to patches of necrosis. Peri-necrotic cells are typically hypoxic and capable of existing at very low oxygen concentrations (PO 2 ≤ 1%) ( Hall and Giaccia, 2006 ). In normal cells, hypoxia typically leads to cell death. Counter-intuitively, however, hypoxia can induce genomic changes that enable tumor cells to adapt to poor nutrition and the hostile microenvironment, thus remaining viable. Consequently, hypoxia exerts a selection pressure that leads to the survival of subpopulations of viable cells with the genetic machinery for malignant progression ( Vaupel and Harrison, 2004 ). Tumors can overcome the proliferation limitations posed by the stressful microenvironment by stimulating the production of new blood vessels via the release of hypoxia-inducible angiogenic factors, such as vascular endothelial growth factor (VEGF) to develop a new blood supply ( Seo et al., 2014 ). Paradoxically, following neovascularization in solid tumor tissue, which consists of poorly organized, elongated, dilated, twisted and blind-ended blood vessels, oxygen supply for the tumor may still be deficient ( Helmlinger et al., 1997 ).
The adaptive immune system relies on different cell types to provide fast and coordinated responses, characterized by recognition of pathogenic challenge, extensive cellular prolifer- ation and differentiation, as well as death. T cells are a subset of the adaptive immune cellular pool that recognize immunogenic peptides expressed on the surface of antigen- presenting cells by means of specialized receptors on their membrane. T cell receptor binding to ligand determines T cell responses at different times and locations during the life of a T cell. Current experimental evidence provides support to the following: (i) sufﬁciently long receptor – ligand engagements are required to initiate the T cell signalling cascade that results in productive signal transduction and (ii) counting devices are at work in T cells to allow signal accumulation, decoding and translation into biological responses. In the light of these results, we explore, with mathematical models, the timescales associated with T cell responses. We consider two different criteria: a stochastic one (the mean time it takes to have had N receptor – ligand complexes bound for at least a dwell time, t, each) and one based on equilibrium (the time to reach a threshold number N of receptor – ligand complexes). We have applied mathematical models to previous experiments in the context of thymic nega- tive selection and to recent two-dimensional experiments. Our results indicate that the stochastic criterion provides support to the thymic afﬁnity threshold hypothesis, whereas the equilibrium one does not, and agrees with the ligand hierarchy experimentally established for thymic negative selection.
Finally, hGFs and hBM-MSCs cultured on NN surfaces showed a high frequency of alignment, in contrast to cells cultured on control Ti surfaces, which showed a random orientation. The increased cell differentiation induced by the NN surfaces observed in both cell types could be explained by this higher frequency of alignment. Previous studies have demonstrated that nanostructured surfaces can induce hBM-MSCs’ osseodifferentation . In addition, it is known that surface topographical features can modulate cell response (adhesion, migration, proliferation, and differentiation) through mechanotransduction [3,4,30]. Specifically, MSCs’ differentiation is improved with a well-organized cytoskeleton and a well-spread cell morphology—features that we observe in cells cultured onto NN surfaces. In our study, we found a lack of effect on cell adhesion by the presence of NN on the surface, but this nanostructuration induced an oriented cell alignment for hGFs and hBM-MSCs, leading to an increased functionality (collagen deposition and ALP activity), indicating that this structure has a positive effect on cell differentiation.
If the location or the Ca 2+ handling properties of mito- chondria in these functional triads could be regulated, this could be an effective mechanism for modulation of the exocytotic process. If these mechanisms could be extrapo- lated to neurons, these changes of these functional triads could contribute to modulation of synaptic plasticity. Final- ly, under brain stress conditions, such as excitotoxicity or ischemia – reperfusion damage, ageing or development of neurodegenerative diseases, mitochondrial dysfunction may reduce the ability of the mitochondria to muffle cyto- solic Ca 2+ , this leading to increased secretion of excitatory neurotransmitters and cell overactivation, a vicious circle that may trigger processes leading to cell death . Changes in CICR could also modulate the synaptic efficacy under physiological or pathophysiological conditions.
Tamarillo, Cyphomandra betacea (Cav.) Sendt., is a solanaceous tree also known by other common names such as tree tomato or ‘tomate de la paz’ (Bois, 1927) depending on the region where it grows. Originating from the Andean region of Peru, Chile, Ecuador and Bolivia it has spread to several other world areas (Dawes and Pringle, 1983; Meadows, 2002) and nowadays New Zealand is the main producer and exporter of this fruit crop. The main economic interest in this species is related to the high nutritional value of its fruits, which can be eaten fresh or processed, and which have a relatively high content of proteins, vitamin C, vitamin E, provitamin A and minerals, such as potassium and iron (McCane and Widdowson, 1992). More recent studies have shown that tamarillo fruits are also rich in anthocyanins and carotenoids and may be used as a source of antioxidants (Hurtado et al., 2009; Kou et al., 2008). In vitro propagation by axillary shoot proliferation (Barghchi, 1986; Cohen and Elliot, 1979), induction of organogenesis (Guimarães et al., 1996; Obando and Jordan, 2001) or somatic embryogenesis (Correia et al., 2009) and genetic transformation techniques (Atkinson and Gardner, 1993; Cohen et al., 2000) are valuable biotechnological tools that have been applied to tamarillo breeding. SE is of particular interest because it has a great potential for the improvement of tree species and for rapid large-scale clonal propagation, genetic transformation, and cryopreservation of desirable selected lines (Park, 2002; von Arnold, 2008). Moreover, somatic embryo induction and development serve as model to understand genetic regulation of embryo development (Rose et al., 2010; Yang and Zhang, 2010).
Intracellular free calcium concentration ([Ca2+]cyt) plays a pivotal role in control of a large variety of cell and physiological functions in most cells and tissues from the very short term (neurotransmitter release, muscle contraction) to the long-term scale (gene expression, cell proliferation). In general, an increase in [Ca2+]cyt triggers cell activation. However, if the rise in [Ca2+]cyt is large and/or sustained enough, it promotes rather cell death in many cell types including neurons . Examples include overly activation of ionotropic glutamatergic receptors during excitotoxicity, particularly N-methyl d -aspartate (NMDA) [2, 3] as well as Ca2+
The death receptor pathway plays an important role in the immune system. For instance, when we become infected by a virus, cytotoxic T lymphocytes are activated and then express CD95 ligand. By binding to its receptor, CD95 ligand induces apoptosis in the infected cells to prevent the virus from replicating and spreading to other cells. Cytotoxic T cells, in addition, utilize a granule-exocytosis pathway for the elimination of virus- infected cells. Cytotoxic granules contain a pore-forming protein, perforin, and serine proteases known as granzymes. Granzyme B can directly cleave proteins after aspartate residues and can therefore also activate caspase-3. In this way the upstream events of death receptor signaling are bypassed, and apoptosis is induced directly (Figure 2). Most cell death, however, proceeds via the mitochondrial pathway, which integrates signals generated by a variety of stressors including DNA damage, loss of adhesion, growth factor withdrawal and others . As a key event these apoptotic stimuli evoke the release of cytochrome c from the mitochondrial intermembrane space into the cytosol. Cytochrome c normally functions in electron transport processes of the respira- tory chain to generate ATP. In the cytosol of apoptotic cells, however, it serves as a cofactor for the adapter protein Apaf-1. Upon binding of cytochrome c, Apaf-1 oli- gomerizes and recruits the initiator caspase-9 to trigger the formation of the apopto- some . In this case the binding of caspase-9 to Apaf-1 is mediated by a shared homotypic interaction motif known as the caspase recruitment domain (CARD). Thus, like the DISC, the apoptosome is a high-molecular weight complex that serves as a caspase activation platform. Once assembled in the apoptosome, caspase-9 becomes activated and subsequently triggers the caspase cascade (Figure 2).
Finally, the technique was evaluated to check its potential when using micro-encapsulation conditions. This time, the active chamber of the resonator, that is the active volume that contains the gel treated with ultrasound, was only 1mm in diameter. The frequency used (1MHz, Fig 5) allowed only one nodal plane to be generated within the field. In this case, auto- fluorescent particles were used to allow confocal microscopy analysis. The image of the sample collected and analysis by light microscopy suggested that the cells re-arranged along the minimum pressure areas, confirming the expectations. Moreover, it was demonstrated clearly that the cell position is dependent on the frequency used. Confocal microscopy image analysis of the gel treated using the third harmonic (3 MHz, Fig 6) of the resonant frequency clearly showed the image of the horizontal section of the areas with the cell distributions. Three concentric circles can be identified coincidental with the three nodal pressure expected.
Angioimmunoblastic T-cell lymphoma (AITL) accounts for 15–20% of all peripheral T-cell lymphomas. It is a rare subtype of CD4 T-cell peripheral lymphoma that affects aged in- dividuals, causing B symptoms, generalized lymphadenopathy and hepatosplenomegaly. Its pathogenesis is still unclear, but in some cases it has been associated with infection, allergic reaction or drug exposure. The majority of patients are diagnosed in an advanced stage and anthracycline based regimen is considered the first-line therapy. Skin involvement is not well characterized, occurring in up to 50% of patients and presenting as nonspecific rash, mac- ules, papules, petechiae, purpura, nodules and urticaria. We present the illustrative case of a 55-year-old woman with an AITL who presented prominent skin findings, arthritis, lymphade- nopathy and hypereosinophilia. Skin biopsy reported a T-cell lymphoma and the diagnosis of AITL was confirmed by an axillary lymph node biopsy, which was also positive for Epstein- Barr virus. Chemotherapy with CHOP-21 and thalidomide was given, accomplishing complete