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This page shows you the most recent publications within this specialty of the MedWorm directory. This is page number 11.

Utilizing a Retroviral RNAi System to Investigate In Vivo mTOR Functions in T Cells
RNA interference (RNAi) is an intracellular mechanism for silencing gene expression utilizing short fragments of double-strand RNA that are complementary to the target messenger RNA. This gene silencing technique has now become an invaluable research tool due to its specific and strong repressive effect on a target transcript. We have recently applied a retrovirus-based RNAi system to investigate the in vivo role of the mammalian target of rapamycin (mTOR) in antigen-specific CD8 T cells, and have found that mTOR regulates memory CD8 T-cell differentiation. Here, we provide a detailed protocol for knocking down mTOR and it...
Source: Springer protocols feed by Cell Biology - December 2, 2011 Category: Cytology Source Type: news

Rapamycin-Induced Enhancement of Vaccine Efficacy in Mice
Th1 immunity protects against tuberculosis infection in mice and humans. The widely used BCG vaccine primes CD4 and CD8 T cells through signaling mechanisms from dendritic cells and macrophages. The latter express MHC-II and MHC-I molecules through which peptides from BCG vaccine are presented to CD4 and CD8 T cells, respectively. Since BCG sequesters within a phagosome that does not fuse with lysosomes, generation of peptides within antigen-presenting cells infected with BCG occurs with reduced efficiency. We demonstrate that activation of DCs containing BCG vaccine with rapamycin leads to an enhanced ability of DC vaccin...
Source: Springer protocols feed by Cell Biology - December 2, 2011 Category: Cytology Source Type: news

Expanding Human T Regulatory Cells with the mTOR-Inhibitor Rapamycin
CD4+CD25+FOXP3+ T regulatory (Treg) cells are pivotal for the induction and maintenance of peripheral tolerance in both mice and humans. The possibility to use Treg cells for the treatment of T-cell-mediated diseases has recently gained increasing momentum. However, given the limited amount of circulating FOXP3+ Treg cells, efficient methods for their ex vivo expansion are highly desirable. Rapamycin allows for in vitro expansion of murine and human FOXP3+ Treg cells, which maintain their regulatory phenotype and suppressive capacity. Here, we describe in detail the powerful methods for enriching human FOXP3+ Treg cells st...
Source: Springer protocols feed by Cell Biology - December 2, 2011 Category: Cytology Source Type: news

Inducible raptor and rictor Knockout Mouse Embryonic Fibroblasts
The mammalian Target of Rapamycin (mTOR) kinase functions within two structurally and functionally distinct multiprotein complexes termed mTOR complex 1 (mTORC1) and mTORC2. The immunosuppressant and anticancer drug rapamycin is commonly used in basic research as a tool to study mTOR signaling. However, rapamycin inhibits only, and only incompletely, mTORC1, and no mTORC2-specific inhibitor is available. Hence, a full understanding of mTOR signaling in vivo, including the function of both complexes, requires genetic inhibition in addition to pharmacological inhibition. Taking advantage of the Cre/LoxP system, we generated ...
Source: Springer protocols feed by Cell Biology - December 2, 2011 Category: Cytology Source Type: news

The In Vivo Evaluation of Active-Site TOR Inhibitors in Models of BCR-ABL+ Leukemia
Preclinical evaluation of candidate anticancer compounds requires appropriate animal models. Most commonly, solid tumor xenograft systems are employed in which immunocompromised mice are implanted with human cancer cell lines. Genetically engineered mouse models of solid tumors are also frequently employed. Both of these approaches can also be applied to studies of hematological malignancies. In this chapter, we describe three types of mouse models of leukemia driven by the human BCR-ABL oncogene. We also discuss the application of these models to preclinical testing of active-site TOR inhibitors, a novel class of compound...
Source: Springer protocols feed by Cell Biology - December 2, 2011 Category: Cytology Source Type: news

Quantitative Visualization of Autophagy Induction by mTOR Inhibitors
Autophagy is a catabolic pathway that degrades bulk cytosol in lysosomal compartments enabling amino acids and fatty acids to be recycled. One of the key regulators of autophagy is the mammalian target of rapamycin (mTOR), a conserved serine/threonine kinase which suppresses the initiation of the autophagic process when nutrients, growth factors, and energy are available. Inhibition of mTOR, e.g., by small molecules such as rapamycin, results in activation of autophagy. To quantify autophagy induction by mTOR inhibitors, we use an mCherry-GFP-LC3 reporter which is amenable to retroviral delivery into mammalian cells, stabl...
Source: Springer protocols feed by Cell Biology - December 2, 2011 Category: Cytology Source Type: news

Assessing Cell Size and Cell Cycle Regulation in Cells with Altered TOR Activity
Target of rapamycin (TOR) regulates the growth of cells and organisms. Numerous growth-promoting and growth-arresting pathways converge on TOR; TOR acts as an important hub, balancing the pro- and anti-growth signals within a cell. Since it regulates growth at the cellular level, cell size can be used as an indirect readout of TOR activity. Here, we describe methods used to analyze cell size in cell culture and in the Drosophila wing.
Source: Springer protocols feed by Cell Biology - December 2, 2011 Category: Cytology Source Type: news

Immunohistochemical Analysis of mTOR Activity in Tissues
mTOR is a key regulator of cell growth and size, and its activity is often dysregulated in a wide variety of diseases. The mTOR signaling pathway is also a therapeutic target for many diseases, including cancer. Immunohistochemistry is a powerful method to assess mTOR activity in clinical/histological samples, however, care should be taken in choosing the targets for determining mTOR activity due to the complexity of its regulation. This chapter describes the most up-to-date methods for visualizing mTOR activity by immunohistochemistry using commercially available antibodies, including considerations for validating new ant...
Source: Springer protocols feed by Cell Biology - December 2, 2011 Category: Cytology Source Type: news

A Genome-wide RNAi Screen for Polypeptides that Alter rpS6 Phosphorylation
Mammalian target of rapamycin (mTOR) is a giant protein kinase that controls cell proliferation, growth, and metabolism. mTOR is regulated by nutrient availability, by mitogens, and by stress, and operates through two independently regulated hetero-oligomeric complexes. We have attempted to identify the cellular components necessary to maintain the activity of mTOR complex 1 (mTORC1), the amino acid-dependent, rapamycin-inhibitable complex, using a whole genome approach involving RNAi-induced depletion of cellular polypeptides. We have used a pancreatic ductal adenocarcinoma (PDAC) cell line, Mia-PaCa for this screen; as w...
Source: Springer protocols feed by Cell Biology - December 2, 2011 Category: Cytology Source Type: news

Evaluation of mTOR-Regulated mRNA Translation
mTOR, the mammalian target of rapamycin, regulates protein synthesis (mRNA translation) by affecting the phosphorylation or activity of several translation factors. Here, we describe methods for studying the impact of mTOR signalling on protein synthesis, using inhibitors of mTOR such as rapamycin (which impairs some of its functions) or mTOR kinase inhibitors (which probably block all functions).To assess effects of mTOR inhibition on general protein synthesis in cells, the incorporation of radiolabelled amino acids into protein is measured. This does not yield information on the effects of mTOR on the synthesis of specif...
Source: Springer protocols feed by Cell Biology - December 2, 2011 Category: Cytology Source Type: news

2 top biological imaging centers offer powerful free online tool to researchers and public
(American Society for Cell Biology) The collaboration of two leading cell image resource centers now provides a more extensive and advanced facility for archiving, sharing, and analyzing microscope images in great detail. The American Society for Cell Biology and the National Center for Microscopy and Imaging Research at the University of California, San Diego, have joined forces to provide a unified interface. This union extends ASCB's The Cell: An Image Library website with new capabilities and an extensible software infrastructure.
Source: EurekAlert! - Medicine and Health - December 1, 2011 Category: Global & Universal Source Type: news

2012 C. elegans Development, Cell Biology, and Gene Expression Meeting
The next C. elegans Development, Cell Biology, and Gene Expression Meeting will be held on the campus of the University of Wisconsin-Madison, June 7-10. Keynote Speakers will include Julie Ahringer, David Greenstein, Ken Kemphues, Craig Mello, Karen Oegema, Shai Shaham and Anne Villeneuve. Organizers: E. Jane Hubbard, Jeremy Nance, Martha Soto. http://www.union.wisc.edu/celegans/index2012.html
Source: WormBase - November 30, 2011 Category: Genetics & Stem Cells Authors: rkishore Tags: meetings news Source Type: news

Sammy Franks obituary
Prostate pathologist whose research linked cancer and ageingLeonard Maurice Franks, fondly known as "Sammy", who has died aged 90, was regarded as the doyen of prostate pathologists. Particularly important was his research into the high frequency of prostate cancer in ageing men, which led to the appreciation that if a man lives long enough, he will develop prostate cancer. Sammy was also a pioneer in the application of cell culture and electron microscopy to cancer research. He was for decades a revered authority on tumour biology.He was born in Newcastle upon Tyne and graduated in medicine from Durham University in 1942....
Source: Guardian Unlimited Science - November 29, 2011 Category: Science Authors: Nicholas Wright Tags: Medical research People in science Health Society The Guardian Obituaries Source Type: news

Recycling Of BACE1 Enzyme Implicated In Promotion Of Alzheimer's Disease
Sluggish recycling of a protein-slicing enzyme could promote Alzheimer's disease, according to a study published online in The Journal of Cell Biology*. Abeta, the toxic protein that accumulates in the brains of Alzheimer's patients, is formed when enzymes cut up its parental protein, known as amyloid precursor protein. One of those enzymes is beta-secretase or BACE1. BACE1 cycles between the Golgi apparatus and the plasma membrane, traveling through endosomes on the way. A protein complex called the retromer helps transport proteins back from endosomes to the Golgi...
Source: Health News from Medical News Today - November 23, 2011 Category: Consumer Health News Tags: Alzheimer's / Dementia Source Type: news

European Union names its first chief scientist
Nature: Anne Glover, a molecular and cell biologist who is currently serving as Scotland's chief scientific adviser, will become the European Union's first chief scientific adviser. Although the identity of the nominee has been announced, key aspects of her job remain undefined, writes Nature's Natasha Gilbert. In particular, it's not clear how much real influence Glover will have over science policy or which EU office she will belong to. The position of chief scientific adviser was created two years ago. Máire Geoghegan-Quinn, the EU research commissioner, blamed the delay in filling the position on the financial crisis.
Source: Physics Today News Picks - November 22, 2011 Category: Physics Authors: Physics Today Source Type: news

At-home cervical screening offers alternative for underserved women
Human papillomavirus testing undertaken at home is more sensitive at detecting invasive cervical cancer than traditional cytology, and could offer a feasible option to women who are unable/unwilling to attend cervical screening, say researchers.
Source: MedWire News - Ob/Gyn - November 17, 2011 Category: OBGYN Source Type: news

Dazzling Miniatures: View Highlights from BioScapes Photo Contest
Microscopy remains one of the few areas of science in which enthusiastic amateurs can make others take notice. Nonprofessionals routinely produce stunning images of creatures and objects too tiny for the eye to resolve. This crowdsourcing of microscopic imagery arrived long before the invention of the smartphone and networked communications: the amateur has long made a mark with the microscope--in the early years, by hand drawing images that appeared underneath the lens, and, in more recent times, with the added realism brought by the photograph.This noble tradition continues in our pages, as we offer a selection of photog...
Source: Scientific American Topic - Biotechnology - November 14, 2011 Category: Biotechnology Tags: More Science,Technology,Society & Policy,Biotechnology,Science Education,Evolutionary Biology,Biotechnology,Biology Source Type: news

At-home cervical screening offers alternative for underserved women
Human papillomavirus testing undertaken at home is more sensitive at detecting invasive cervical cancer than traditional cytology, and could offer a feasible option to women who are unable/unwilling to attend cervical screening, say researchers.
Source: MedWire News - Oncology - November 13, 2011 Category: Cancer & Oncology Source Type: news

Evaluation of Cytochrome P450 Activities in Human Hepatocytes In Vitro
Major hepatic cytochrome P450 activities (CYP1A2, CYP2A6, CYP2B6, CYP2C9, CYP2C19, CYP2D6, CYP2E1, and CYP3A4) can be simultaneously examined in human hepatocytes by incubation with a cocktail of multiple specific probes. Cocktail strategy in combination with mass spectrometry is shown to be a robust, fast, and sensitive procedure for P450 activity assessment. This procedure allows a drastic reduction of the number of cells required in the assay and sample analysis time and increases throughput and reproducibility. Major applications of the probe cocktail strategy are P450 phenotyping of hepatocytes and induction studies.
Source: Springer protocols feed by Cell Biology - November 11, 2011 Category: Cytology Source Type: news

Conversion of Non-endocrine Human Pancreatic Cells to Insulin-Producing Cells for Treatment of Diabetes
Type I diabetes results from the autoimmune destruction of the insulin-secreting pancreatic β-cells, affecting many millions of people worldwide. The optimal treatment is to restore the endogenous supply of insulin either through the transplantation of pancreas or the transplantation of islets of langerhans or simply the β-cells. However, the donated pancreas organs are limited and the available organs are only able to treat a small portion of the diabetes patients. Thus, glucose-responsive, insulin-producing cells from human origin are urgently needed. The aim of this chapter is to give some insight views to how...
Source: Springer protocols feed by Cell Biology - November 11, 2011 Category: Cytology Source Type: news

Functional Analysis of Human Islets of Langerhans Maintained in Culture
Islets of Langerhans isolated from experimental animals, such as mice and rats, have contributed much to our understanding of the mechanisms by which pancreatic β-cells secrete insulin in a regulated manner, and this knowledge is important in identifying potential novel therapies for Type 2 diabetes. However, although many of the signal transduction pathways identified in rodent islets are common to humans, some critical differences have been demonstrated experimentally. It is, therefore, essential that experiments are performed using islets isolated from human pancreas to provide robust data defining whether the key ...
Source: Springer protocols feed by Cell Biology - November 11, 2011 Category: Cytology Source Type: news

Culture of Parathyroid Cells
The parathyroid cells are highly differentiated with more or less their only function to secrete parathyroid hormone in response to the extracellular calcium level. Tumours from the parathyroid glands are >99% benign, and have a slow proliferation rate. Culture of parathyroid cells is known to be very difficult most likely due to the high differentiation level. This chapter reveals some details in order how to get parathyroid cells to survive in culture after dispersion of normal bovine parathyroid glands or pathological human parathyroid tumours. Detailed protocols describing cell dispersion with collagenase, short-ter...
Source: Springer protocols feed by Cell Biology - November 11, 2011 Category: Cytology Source Type: news

Isolation, Cultivation, and Application of Human Alveolar Epithelial Cells
The blood–air barrier formed by the alveolar epithelium of the peripheral lung is crucial for the pulmonary delivery of drugs. Most existing in vitro models mimicking the blood–air barrier are represented by tumor cells or immortalized cells and lack biological relevance due to their genetic alterations and underexpressed essential physiological functions. However, the increasing interest of aerosol administration of medicines to the respiratory system requires the development and use of representative in vitro models. Thereby, human alveolar epithelial cells (hAEpC) are a suitable test system allowing standard...
Source: Springer protocols feed by Cell Biology - November 11, 2011 Category: Cytology Source Type: news

Isolation and Cultivation of Dermal Stem Cells that Differentiate into Functional Epidermal Melanocytes
Human melanocytes have been extensively studied, but a melanocyte stem cell reservoir in glabrous skin has not yet been found. Human dermis contains cells that are nonpigmented but can differentiate to several different cell types. We have recently shown that multipotent dermal stem cells isolated from human neonatal foreskins are able to differentiate to multiple cell lineages, including pigmented melanocytes. The dermal stem cells grow as three-dimensional spheres in human embryonic stem cell medium and express some neural crest stem cell and embryonic stem cell markers. Melanocytes derived from dermal stem cells express...
Source: Springer protocols feed by Cell Biology - November 11, 2011 Category: Cytology Source Type: news

Culturing and Differentiating Human Mesenchymal Stem Cells for Biocompatible Scaffolds in Regenerative Medicine
Mesenchymal stem cells from a variety of sites are a natural resource that using appropriate skills can be cultured in the laboratory, in scaffolds, to provide differentiated-cell replacement tissues, for clinical application. To perform such work with human cells, strict ethical integrity must be observed at all stages. Adipocytes, osteocytes and chrondrocytes are amongst the most desirable end-point cells. Hydrolytic degradable scaffolds allow implanted cells to synthesise their own extracellular matrix in situ after implantation, degeneration of the foreign scaffold to temporally match creation of the new innate one. Fo...
Source: Springer protocols feed by Cell Biology - November 11, 2011 Category: Cytology Source Type: news

Automated Adherent Human Cell Culture (Mesenchymal Stem Cells)
Human cell culture processes developed at research laboratory scale need to be translated to large-scale production processes to achieve commercial application to a large market. To allow this transition of scale with consistent process performance and control of costs, it will be necessary to reduce manual processing and increase automation. There are a number of commercially available platforms that will reduce manual process intervention and improve process control for different culture formats. However, in many human cell-based applications, there is currently a need to remain close to the development format, usually a...
Source: Springer protocols feed by Cell Biology - November 11, 2011 Category: Cytology Source Type: news

Laser Microdissection Microscopy: Application to Cell Culture
Laser microdissection (LMD) microscopy allows isolation of specific cell populations to target their molecular profile. There are several different types of LMD microscopes, but they are all based on the same principle. A laser beam is used to cut out cells or tissues of interest from a histological section, cytology preparations, or live cells from tissue cultures. Live cells can be isolated using LMD and processed for downstream molecular work. RNA, DNA, and protein isolation is possible from a small number of cells and the material is suitable for further real-time PCR, ELISA, Western Blotting, and protein microarray analysis.
Source: Springer protocols feed by Cell Biology - November 11, 2011 Category: Cytology Source Type: news

Effects of Temperature Generated from the Holmium: YAG Laser on Human Osteoblasts in Monolayer Tissue Culture
With the use of lasers for ablation purposes in spinal surgery, the tissue temperature increases above the boiling point of water, leading to tissue ablation by vaporisation. Due to the thermal environment engendered by the use of lasers, there is concern about the safety of the surrounding important structures, such as dura mater, dorsal root ganglia, and nerve roots.
Source: Springer protocols feed by Cell Biology - November 11, 2011 Category: Cytology Source Type: news

Human Osteoclast Culture and Phenotypic Characterization
Bone remodelling occurs throughout life via the coupled actions of bone resorption and bone formation. When the balance of bone resorption and formation becomes unequal, bone diseases, such as osteoporosis occur, while the absence of functional osteoclasts leads to diseases such as osteopetrosis and pycnodysostosis. In order to develop effective treatments for bone disease the normal regulatory systems involved in bone resorption need to be fully elucidated. The only cell in the body capable of resorbing bone is the osteoclast – a highly specialized cell of haematopoietic origin. Until relatively recently, the abilit...
Source: Springer protocols feed by Cell Biology - November 11, 2011 Category: Cytology Source Type: news

Isolation and Culture of Human Osteoblasts
The skeleton is a dynamic organ that is constantly active throughout life. The highly coordinated actions of bone cells early in life determine the body’s shape and form, whilst the constant remodelling (bone resorption followed by an equal amount of bone formation) during adulthood helps to maintain skeletal mass and repair microdamage. When the balance of bone resorption and bone formation becomes unequal, bone diseases, such as osteoporosis, occur. In order to develop drugs to combat bone disease, it is important to know the regulatory systems involved in normal bone formation and resorption. In this chapter, we c...
Source: Springer protocols feed by Cell Biology - November 11, 2011 Category: Cytology Source Type: news

Human Chondrocyte Cultures as Models of Cartilage-Specific Gene Regulation
The human adult articular chondrocyte is a unique cell type that has reached a fully differentiated state as an end point of development. Within the cartilage matrix, chondrocytes are normally quiescent and maintain the matrix constituents in a low-turnover state of equilibrium. Isolated chondrocytes in culture have provided useful models to study cellular responses to alterations in the environment such as those occurring in different forms of arthritis. However, expansion of primary chondrocytes in monolayer culture results in the loss of phenotype, particularly if high cell density is not maintained. This chapter descri...
Source: Springer protocols feed by Cell Biology - November 11, 2011 Category: Cytology Source Type: news

Human CD4+CD25highCD127low/neg Regulatory T Cells
CD4+CD25highCD127low/neg regulatory T cells (Tregs) play a critical role in the maintenance of peripheral tolerance and in controlling the development of autoimmune diseases. A combination of surface and intracellular markers, namely, CD25, CD39/CD73, CD62L, CD45RO, CD127, glucocorticoid-induced tumor necrosis factor receptor (GITR), CTLA-4, and the forkhead/winged helix transcription factor (FOXP3), has been used to characterize Tregs. Tregs suppress T effector responses mainly in a direct cell–cell contact manner. However, other mechanisms independent from this manner cannot be excluded entirely. It has been shown ...
Source: Springer protocols feed by Cell Biology - November 11, 2011 Category: Cytology Source Type: news

Introduction to Cell Culture
The basics of cell culture as applied to human cells are discussed. Biosafety when working with human tissue, which is often pathogenic, is important. The requirements for a tissue culture laboratory are described, particularly the range of equipment needed to carry out cell isolation, purification, and culture. Steps must be taken to maintain aseptic conditions to prevent contamination of cultures with micro-organisms. Basic cell-handling techniques are discussed, including choice of media, primary culture, and cryopreservation of cells so they can be stored for future use. Common assays which are used to determine cell v...
Source: Springer protocols feed by Cell Biology - November 11, 2011 Category: Cytology Source Type: news

Human Peripheral Blood Mononuclear Cell Culture for Flow Cytometric Analysis of Phosphorylated Mitogen-Activated Protein Kinases
Lymphocyte activation and fine tuning of downstream signaling circuits for the regulation of cytokine expression are critical for a successful immune response. Hence, technical protocols permitting simultaneous testing of these attributes in peripheral blood lymphocytes are of paramount importance. Phospho-specific flow cytometry is a novel methodology that detects phosphorylation of signaling effectors in multiple, rare cellular populations within peripheral blood. In addition, it allows the quantification of phosphorylation levels for signaling proteins within each single cell, and therefore is superior compared to tradi...
Source: Springer protocols feed by Cell Biology - November 11, 2011 Category: Cytology Source Type: news

Culture of Human Endothelial Cells from Umbilical Veins
The present protocol offers an economical option for the isolation and culture of human endothelial cells for vascular cell biology research due to the non-invasive collection procedure being devoid of ethical concerns and ease of the isolation technique, expansion and maintenance under standard cell culture conditions. The human umbilical vein endothelial cell (HUVEC) model is useful for any research on general properties of human endothelium, but as these cells are of foetal and venous origin, other sources could be more appropriate models for studies on specific pathological areas, for example, atherosclerosis or cancer...
Source: Springer protocols feed by Cell Biology - November 11, 2011 Category: Cytology Source Type: news

Human Vascular Smooth Muscle Cell Culture
Human vascular smooth muscle cells (VSMCs) in culture are an important tool in understanding how VSMCs function and contribute to vessel wall contraction as well as disease. In this chapter, we describe methodologies that enable the investigator to culture large numbers of proliferative VSMCs. These VSMCs are heterogeneous and vary in size, shape, and proliferative capacity depending on the disease state and location of the vessel of origin. Therefore, we also describe techniques to validate their identity as bone fide VSMCs. Briefly, the methods include information on how to dissect the blood vessel to remove the medial l...
Source: Springer protocols feed by Cell Biology - November 11, 2011 Category: Cytology Source Type: news

Primary Culture of Ovarian Cells for Research on Cell Interactions in the Hormonal Control of Steroidogenesis
Ovary is a polymorphic complex structure in which the cells are arranged in two essential endocrine mini glands: the follicle (F) and the corpus luteum (CL). Their secretory function creates an optimal milieu for growth, maturation, and selection of follicles and oocytes competent for ovulation. Monoculture of isolated ovarian cells has identified the secretory potential of the different cell types functioning in this complex gland in vivo. Primary culture of isolated ovarian cells is a good tool for the investigation of cell interactions and its impact on steroidogenesis, dynamics of steroidogenic enzymes, hormone recepto...
Source: Springer protocols feed by Cell Biology - November 11, 2011 Category: Cytology Source Type: news

Primary Culture of Human Adipocyte Precursor Cells: Expansion and Differentiation
Culture of adipose tissue precursor cells allows gaining insight into the sequential processes involved in adipocyte development. Furthermore, the secretory properties associated with these cellular changes can be studied. Although clonal cell lines are valuable tools for the identification of mechanisms associated with proliferation or differentiation such models do not necessarily represent the complexity of adipose tissue physiology. Primary cell culture systems may be closer to physiology and circumvent some of these restrictions. One advantage is that phenotypic properties of the tissue donor such as gender, age, or b...
Source: Springer protocols feed by Cell Biology - November 11, 2011 Category: Cytology Source Type: news

Culture of Isolated Human Adipocytes and Isolated Adipose Tissue
Adipose tissue (AT) is no longer considered merely as insulation or padding for human organs. It is an endocrine organ in its own right, which includes composite cells with the ability to differentiate into multiple cell lines. In fact, there is increasing evidence to support the theory that the causation of obesity and its associated metabolic disorders originate at the cellular or tissue level. Adipocyte dysfunction and chronic inflammatory states are able to modulate triglyceride storage and mobilization directly through cytokine and adipokine release. Significant variability exists between adipocyte isolation and cultu...
Source: Springer protocols feed by Cell Biology - November 11, 2011 Category: Cytology Source Type: news

Glomerular Epithelial and Mesangial Cell Culture and Characterization
Culture of isolated kidney glomerular cells has been employed for almost four decades as a tool to dissect pathophysiological effects of individual cell types in renal disease. This chapter aims to highlight in detail the available techniques to isolate, culture, and characterize human glomerular epithelial and mesangial cells. To establish primary culture of these cells, glomeruli are isolated from the cortex of kidney by differential sieving and cellular outgrowths from cultured glomeruli further subcultured in appropriately coated tissue culture plates/flasks. Methods used for characterization of isolated glomerular mes...
Source: Springer protocols feed by Cell Biology - November 11, 2011 Category: Cytology Source Type: news

Primary Culture of Human Renal Proximal Tubule Epithelial Cells and Interstitial Fibroblasts
Renal physiology and pathology are complex systems that are best studied in whole living organisms. This, however, is often restricted by our desire to limit the number of animal experiments undertaken and to replace them with relevant in vitro models that can be used as surrogates for the system under test. Primary culture cells are derived directly from the relevant tissue and therefore correlate more closely with the system under examination. Although the tissue of origin is not always readily available for culture and cells may quickly change their phenotype after only a few passages, they can be used in many circumsta...
Source: Springer protocols feed by Cell Biology - November 11, 2011 Category: Cytology Source Type: news

Isolation, Characterization, and Culture of Normal Human Intestinal Crypt and Villus Cells
The intestinal epithelium is a highly dynamic tissue undergoing constant and rapid renewal. It consists of a functional villus compartment responsible for terminal digestion and nutrient absorption and a progenitor cell compartment located in the crypts that produce new cells. The mechanisms regulating cell proliferation in the crypt, their migration, and differentiation are still incompletely understood. Until recently, normal human intestinal cell models allowing the study of these mechanisms have been lacking. In our laboratory, using fetal human intestines obtained at mid-gestation, we have generated the first normal h...
Source: Springer protocols feed by Cell Biology - November 11, 2011 Category: Cytology Source Type: news

Isolation and Functional Studies of Human Fetal Gastric Epithelium in Primary Culture
Our understanding of gastric epithelial physiology in man is limited by the absence of normal or appropriate cancer cell lines that could serve as an in vitro model. Research mostly relied on primary culture of gastric epithelial cells of animal species, enriched with surface mucous cells, and devoid of glandular zymogenic chief cells. We successfully applied a new nonenzymatic procedure using Matrisperse Cell Recovery Solution to dissociate the entire epithelium from human fetal stomach. Cultures were generated by seeding multicellular aggregates prepared by mechanical fragmentation. We further demonstrate that this simpl...
Source: Springer protocols feed by Cell Biology - November 11, 2011 Category: Cytology Source Type: news

Mass Spectrometry-Based Chemoproteomic Approaches
The term “chemical proteomics” refers to a research area at the interface of chemistry, biochemistry, and cell biology that focuses on studying the mechanism of action of bioactive small molecule compounds, which comprises the mapping of their target proteins and their impact on protein expression and posttranslational modifications in target cells or tissues of interest on a proteome-wide level. For this purpose, a large arsenal of approaches has emerged in recent years, many of which employing quantitative mass spectrometry. This review briefly summarizes major experiment types employed in current chemical proteomics research.
Source: Springer protocols feed by Protein Science - November 11, 2011 Category: Biochemistry Source Type: news

Two 4-year end-of-study analyses evaluating Cervarix against grade 3 or greater cervical intraepithelial neoplasia and other non-vaccine oncogenic HPV types
Source: Lancet Oncology Area: News The Lancet Oncology has featured two 4-year end-of-study analyses early online in which the efficacy of the HPV-16/18 adjuvanted vaccine was evaluated for: . CIN3+ and adenocarcinoma in situ (AIS) in the PApilloma TRIal against Cancer In young Adults (PATRICIA trial), and . Outcomes of other non-vaccine oncogenic HPV types [Cervical intraepithelial neoplasia grade 2 or greater (CIN2+) is the surrogate endpoint used in licensure trials of human papillomavirus (HPV) vaccines. Vaccine efficacy against CIN3+, the immediate precursor to invasive cervical canc...
Source: NeLM - News - November 9, 2011 Category: Drugs & Pharmacology Source Type: news

Critical Protein Prevents Secondary Damage After Stroke
One of two proteins that regulate nerve cells and assist in overall brain function may be the key to preventing long-term damage as a result of a stroke, the leading cause of disability and third leading cause of death in the United States. In a recent study published in the Journal of Neuroscience, Bonnie Firestein, professor of cell biology and neuroscience, in the School of Arts and Sciences, says the new research indicates that increased production of two proteins - cypin and PSD-95 - results in very different outcomes...
Source: Health News from Medical News Today - November 4, 2011 Category: Consumer Health News Tags: Stroke Source Type: news

Improved Understanding Of Batten Disease Offers A Potential Treatment
Waste management is a big issue anywhere, but at the cellular level it can be a matter of life and death. A Weizmann Institute study, published in the Journal of Cell Biology, has revealed what causes a molecular waste container in the cell to overflow in Batten disease, a rare but fatal neurodegenerative disorder that begins in childhood. The findings may form the basis for a therapy for this disorder. In Batten disease, an insoluble yellow pigment accumulates in the brain's neurons, causing these cells to degenerate and ultimately die...
Source: Health News from Medical News Today - November 4, 2011 Category: Consumer Health News Tags: Neurology / Neuroscience Source Type: news

Frozen Section Library: Lymph Nodes
series:Frozen Section LibraryThe Frozen Section Library series provides concise, user-friendly, site specific handbooks that are well illustrated and highlight the pitfalls, artifacts and differential diagnosis issues that arise in the hurried frozen section scenario. Frozen Section Library: Lymph Nodes highlights the usefulness of techniques such as touch preparation cytology and fine needle aspiration (FNA) cytology, as ...
Source: Springer Medicine titles - November 3, 2011 Category: Databases & Libraries Tags: Pathology Source Type: news

HPV Self-Collected Testing Promising for Developing NationsHPV Self-Collected Testing Promising for Developing Nations
Self-sampling for human papillomavirus (HPV) detected more high-grade lesions and cervical cancers than cytology among low socioeconomic women in Mexico. Medscape Medical News
Source: Medscape Today Headlines - November 3, 2011 Category: Consumer Health News Tags: Hematology-Oncology News Source Type: news

Gene Mutation Associated With Rare Eye Disease Also Contributes To Bladder Cancer Growth
Research conducted by Dr. Jayne S. Weiss, Professor and Chair of Ophthalmology at LSU Health Sciences Center New Orleans, and colleagues has found that a defect in a gene involved in a rare disease of the cornea also contributes to the progression of invasive bladder cancer. The findings are published in the November 2011 issue of the peer-reviewed journal, DNA and Cell Biology. It is the featured research of the issue, selected for the cover. Because earlier studies, including Dr...
Source: Health News from Medical News Today - November 3, 2011 Category: Consumer Health News Tags: Cancer / Oncology Source Type: news