Abstract

Insulin-dependent diabetes mellitus (IDDM) occurs as a consequence of autoimmune destruction of the insulin-producing pancreatic -cells. Although progress has been made in the field of islet transplantation, an appealing alternative strategy for -cell replacement therapy for IDDM is to target insulin expression to non-islet cells. We have recently generated transgenic nonobese diabetic (NOD) mice in which insulin gene expression was targeted to proopiomelanocortin (POMC)-expressing cells of the intermediate lobe (IL) of the pituitary. We have shown that POMC-expressing IL pituitary cells secreted large amounts of mature insulin, similar to islet -cells. However, in contrast to the insulinproducing islet -cells, the insulin-producing IL pituitary cells were not attacked by the immune system. Remarkably, transplantation of small amounts of the transgenic IL tissues into diabetic NOD mice resulted in the restoration of near-normogylcemia and the complete reversal of diabetic symptoms. In separate experiments, IL allografts showed enhanced viability and were highly vascularized, compared with similarly transplanted islet allografts. These features are highly advantageous in the transplantation setting and demonstrate the considerable potential of these nonislet cell types for insulin-gene delivery in IDDM. Key words Insulin gene expression - Intermediate pituitary - Diabetes - Transgenic NOD mice Received: 20 December 1996 / Accepted in revised form: 24 December 1996

(http://www.springerlink.com/content/55lwraccd9qlxjat/)

Despite many triumphs, a significant limitation of the usefulness of many of the available Bcell lines for the study of insulin secretion are either inappropriate or lack of responsiveness to glucose. Commonly employed cell lines generated prior to the 1990s following X-ray irradiation (RINm5F cells) or simian virus 40 B-cell transformation (HIT-T15 cells and BTC) fall into this category. More recent success has been achieved with the generation of INS-1 cells and MIN6 cells, but the production of these cell lines owes much to good fortune, dedication and hard work. In the present era, molecular biology techniques provide the opportunity to engineer novel pancreatic B-cell lines which possess many attributes of normal insulinsecreting cells. This review describes the electrofusion of normal NEDH rat pancreatic B-cells with immortal RINm5F cells to create three new glucose-responsive clonal insulin-secreting cells, designated BRIN-BG5, BRIN-BG7 and BRIN-BD11. These cell lines exhibit up to four-fold insulin-secretory responses to depolarization with 25 mmol/l K+, 7.68 mmol/l Ca2+, 10 mmol/l L-alanine, and activation of protein kinase C or adenylate cyclase with 10 nmol/l phorbol-12myristate-13-acetate or 25 mol/l forskolin, respectively. The maximal insulin-secretory response of both BRIN-BG5 and BRIN-BG7 cells to glucose occurred at 8.4 mmol/l (1.9- and 1.8-fold increases, respectively). In contrast, 4.216.7 mmol/l glucose evoked a stepwise 2- to

3-fold of insulin release from BRIN-BD11 cells. The superior glucose responsiveness of BRINBD11 cells compared with BRIN-BG5 or BRIN-BG7 cells was associated with increased expression of GLUT-2 and a greater contribution of glucokinase to total glucose phosphorylating enzyme activity. Furthermore, BRIN-BD11 cells also showed appropriate responses to a diverse range of modulators of pancreatic B-cell function, including amino acids, neurotransmitters and sulphonylurea drugs. Collectively these observations indicate that genetic modification of insulin-secreting cells by electrofusion (or transfection with cDNA) offers a new avenue for generation of useful clonal glucose-responsive pancreatic Bcell lines for studies of insulin secretion and transplantation in insulin-dependent diabetes mellitus. (http://www.springerlink.com/content/8cwf1t3ujkgwk1ag/ )

Improved Wool Production in Transgenic Sheep Expressing Insulin-like Growth Factor 1 Sami Damak1, Hung-yi Su1, ,*, Nigel P. Jay2 & David W. Bullock1, ,2, ,*

Abstract Transgenic sheep were produced by pronuclear microinjection with a mouse ultrahigh-sulfur keratin promoter linked to an ovine insulin-like growth factor 1 (IGF1) cDNA. Five transgenic lambs resulted from the microinjection of 591 embryos; one male and one female showed IGF1 expression in the skin. A progeny test of the ram was carried out by matings to 43 non-transgenic ewes. Of 85 lambs born, 43 (50.6%) were transgenic. At yearling shearing (approximately 14 months of age), clean fleece weight was on average 6.2% greater in transgenic animals than in their non-transgenic half-sibs, with a greater effect in males (9.2%) than females (3.4%). Transgenics showed a small but significant increase in bulk, but male transgenics had a lower staple strength than female transgenics and nontransgenics which did not differ significantly. There were no significant differences in fiber diameter, medullation, and hogget body weight. To our knowledge this is the first reported improvement in a production trait by genetic engineering of a farm animal without adverse effects on health or reproduction. (http://www.nature.com/nbt/journal/v14/n2/abs/nbt0296-185.html)

Methods :
An o-Toluidine Method for Body-Fluid Glucose Determination
1. Kurt M. Dubowski1 + Author Affiliations 1.
1

Clinical Chemistry and Toxicology Laboratories, University of Florida Teaching Hospital and Clinics, Gainesville, Fla.

Abstract
o-Toluidine, 6% (v/v) in glacial acetic acid, is used to determine glucose in biologic material after deproteinization with 3% (w/v) trichloracetic acid. A stable green color develops after heating at 100 for 10 min., and the absorbance is determined at 630 or 635 m. The reagent is stable for many months at room temperature, and the reaction follows Beer's Law over a very wide range of concentrations. The development of the procedure is discussed, as is the specificity of the method for glucose.

(http://www.clinchem.org/content/8/3/215.short )

Principles of gn

Pancreatic endoderm derived from human embryonic stem cells generates glucose-responsive insulin-secreting cells in vivo
Evert Kroon1, Laura A Martinson1, Kuniko Kadoya1, Anne G Bang1, Olivia G Kelly1, Susan Eliazer1, Holly Young1, Mike Richardson1, Nora G Smart1, Justine Cunningham1, Alan D Agulnick1, Kevin A D'Amour1, Melissa K Carpenter1 & Emmanuel E Baetge1

Abstract
Development of a cell therapy for diabetes would be greatly aided by a renewable supply of human -cells. Here we show that pancreatic endoderm derived from human embryonic stem (hES) cells efficiently generates glucose-responsive endocrine cells after implantation into mice. Upon glucose stimulation of the implanted mice, human insulin and C-peptide are detected in sera at levels similar to those of mice transplanted with ~3,000 human islets. Moreover, the insulin-expressing cells generated after engraftment exhibit many properties of functional -cells, including expression of critical -cell transcription factors, appropriate processing of proinsulin and the presence of mature endocrine secretory granules. Finally,

in a test of therapeutic potential, we demonstrate that implantation of hES cellderived pancreatic endoderm protects against streptozotocin-induced hyperglycemia. Together, these data provide definitive evidence that hES cells are competent to generate glucoseresponsive, insulin-secreting cells.

(http://www.nature.com/nbt/journal/v26/n4/full/nbt1393.html )

Adntages eg :
Title Advantages of genetically modified canola: a Canadian perspective. Authors Devine, M. D.; Buth, J. L. Book chapter; Conference paper The BCPC Conference: Weeds, 2001, Volume 1 and Volume 2. Proceedings of an international conference held at the Brighton Hilton Metropole Hotel, Brighton, UK, 12-15 November 2001 2001 pp. 367-372 Conference Title The BCPC Conference: Weeds, 2001, Volume 1 and Volume 2. Proceedings of an international conference held at the Brighton Hilton Metropole Hotel, Brighton, UK, 12-15 November 2001. ISBN 1-901396-61-4 Record Number 20023047831

Abstract
Herbicide-tolerant (HT) rape (Brassica napus var. oleifera) was introduced commercially in Canada in 1995. Since then, canola varieties or hybrids resistant to glufosinate, glyphosate, imidazolinones or bromoxynil have been introduced. These HT varieties/hybrids have grown in market share such that they now occupy ~70% of the seeded area in western Canada. Growers have adopted HT rape for several reasons, including easier and better weed control, higher seed yield, and higher financial returns and profits, based primarily on the higher yield, reduced dockage and lower herbicide costs. Significant benefits also include reduced herbicide and fuel use. Although growing HT rape does involve critical management decisions, including how best to fit these cultivars into typical crop rotations and management of crop volunteers in subsequent years, their widespread adoption, sustained after 6 years, strongly suggests a net benefit to producers.

Smart and geneticallyengineered biomaterials and drug delivery systems

Jindich Kopeek

Department of Pharmaceutics and Pharmaceutical Chemistry, University of Utah, Salt Lake City, UT 84112, USA

Received 9 January 2003. Revised 9 June 2003. Accepted 10 June 2003. Available online 10 September 2003.

http://dx.doi.org/10.1016/S0928-0987(03)00164-7, How to Cite or Link Using DOI Cited by in Scopus (143)

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Abstract
The design, synthesis, and properties of novel stimuli-sensitive and geneticallyengineered biomaterials and drug delivery systems are reviewed. Two approaches to their engineering are presented. One approach is to improve the traditional methods of synthesis, as demonstrated by the example of controlled copolymerization of -amino acid N-carboxyanhydrides. The other approach, discussed in more detail, uses genetic engineering methods. The design of hybrid hydrogel systems whose components derive from at least two distinct classes of molecules, e.g., synthetic macromolecules and protein domains, is assessed. The design of self-assembling block copolymers is discussed in detail. Finally, the pharmaceutics related applications of these materials are presented. http://www.sciencedirect.com/science/article/pii/S0928098703001647 (

http://books.google.com.qa/books?hl=ar&lr=&id=SqEObv4UBqkC&oi=fnd&pg=PR11&dq=genetic+engineering+an d+insulin+research&ots=45g-WUeJtj&sig=So6fieslF2KpUq8ugcaPUUGbyQ&redir_esc=y#v=onepage&q=genetic%20engineering%20and%20insulin%20research&f=f alse