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BMSG-Biomedical Solutions Group,Inc.-Executive Summaries Houston Medical Center, Texas, USA- College of Medicine Houston-Chicago-Krakow-Poland, September-2016

1. Chondroitin sulphate for treatment of degenerative joint diseases

Chondroitin sulphate is a major constituent of cartilage, providing structure, holding water and nutrients, and allowing other molecules to move through cartilage. In degenerative joint disease, such as osteoarthritis, there is a loss of chondroitin sulphate as the cartilage erodes. The main goal of the project is preparation of chondroitin sulphate from bacterial source. Modulation of transgene expression regulation by introduction of additional regulatory elements and gene modifications is planned. Elaboration of the effective method of chondroitin sulphate purification produced in vivo is also planned. Genetic construct will be composed of specific promoter, modification making genetic construct unique for purpose of patenting and human chondroitin synthase gene.

For atherosclerosis, researchers have sometimes started therapy using very high amounts, such as 5 grams twice per day with meals, lowering the amount to 500 mg three times per day after a few months. Before taking such high amounts, people should consult a doctor. For osteoarthritis, a typical level is 400 mg three times per day. Oral chondroitin sulphate is rapidly absorbed in humans when it is dissolved in water prior to ingestion. Approximately 12% of chondroitin sulphate taken by mouth becomes available to the joint tissues from the blood. Taking into account large population of patients suffering from degenerative joint diseases chondroitin sulphate produced in bacteria may overtake large proportion of pharmaceutical market. Possible use of product on pharmaceutical market.
Last minute info: Gene construct for production in bacteria has been prepared.

2. Production of human growth hormone -HGH in milk of transgenic rabbits

There is strong demand for growth hormone and pharmaceutical applications expanded in last years. The substantial cost limits dramatically utilization of growth hormone. The primary reason for cost relates to complexity of recombinant DNA technology and inherited costs of this technology. One rabbit female can produce up to 10 liters of milk per year. Assuming that expected expression is about 1-10 g/l, a rabbit colony of few hundred animals can supply substantial amount of hormone. The scope of project involves creation of transgenic rabbits producing in mammary gland human growth hormone, achieving stability of hormone expression, high quality of hormone and availability in large quantities.

Mammary gland is considered as the best available bioreactor. Extensive studies have shown that it can serve as the source of a variety of complex recombinant proteins The transgene WAP:6xHishGH became stable integrated into the host genome, can be transmitted to the offspring and expressed. Mammary epithelial cells, which are the source of milk specific protein in the mammary gland, were confirmed to express the transgene.

There is strong demand for growth hormone and pharmaceutical applications expanded in last years. The substantial cost limits dramatically utilization of growth hormone.

The primary reason for cost relates to complexity of recombinant DNA technology and inherited costs of this technology. One rabbit female can produce up to 10 liters of milk per year.

Last minute info: Female rabbits producing growth hormone are available.

3. Transgenic pigs for xenotransplantation

There is a worldwide shortage of organs for clinical transplantation and many patients due to receive new organs die on the waiting list. It is reasonable to consider that organs from other species, may soon be engineered to minimize the risk of serious rejection and used as an alternative to human tissues, possibly ending organ shortages. For most purposes, xenotransplantation will use pigs as the source of tissues and organs. There are ethical, safety and husbandry difficulties in exploiting primates, whereas pigs have large litters and grow to the appropriate size. Xenotransplantation could provide a supply of cells, tissues and organs to treat a number of serious human diseases. The challenge in xenotransplantation is to find a way to persuade the human body's immune system to
accept an animal organ. Xenotransplantation comprises of several important steps from interdisciplinary areas. The first step involves preparation of competitive, inactivating and regulatory gene constructs. These gene constructs should allow (i) knock-out of 1,3 galactosyltransferase (1,3GT) gene, (ii) expression of complement proteins and (iii) regulate expression of proteins.

Potentially, xenotransplantation could provide enough healthy organs to offer transplantation to thousands of patients on waiting lists worldwide. The pig has been selected as the most suitable species for the development of xenotransplantation. The pig's organs are approximately the same size as human organs, both in infancy and adulthood. Additionally, pigs have been domesticated for many centuries. They breed relatively quickly with large litters, so a large number of life-saving organs could potentially be generated quickly when necessary. Generation of transgenic pigs could prevent hyperacute immunological response and make possible use for xenotransplantation.
Last minute info: Homozygous pigs with human 1,2-fucosyltraneserase gene were obtained. Three patent applications filed.

4. Somatic cloning of farm and companion animals

Somatic cloning has been appreciated as the most precise and reliable method of multiplication of desired animal’s features. One of the key applications of the method is in production of transgenic animals. The most promising approach is somatic cloning through nuclear transfer and embryo reconstruction. Methodology optimization for several species (goats, cattle, pigs, rabbits, domestic cats) has crucial importance. Somatic cloning procedure consists of several steps including establishment of somatic cell-lines, evaluation and selection of cells, nuclei transfer, culture of cloned embryos. The aim of project is to optimize each step of the procedure and tune it in the way that supports entire process. Ultimate goal of our project is to reach ability to offer somatic cloning of various farm and domestic animals as specific service with defined, stable and reliable success ratios. Novel method of “blastomere” cloning is developed. This new and unique approach confirmed by experiments performed in rabbits shall grant higher level of cloning safety and efficiency compared to methods developed so far.

Last minute info: Novel cloning procedure has been developed and checked on rabbits. Healthy offspring were born.

5. Flaxseed as a functional food. Bioflax®

Flax has historically been valued for its abundance of fat, which provides a unique mix of fatty acids. Flax is making its mark in the world’s food supply as a functional food. Functional foods deliver a health boost beyond what might be expected from their traditional nutrient content. Flax meets this description perfectly, being rich in alpha-linolenic acid, the essential omega-3 fatty acid, and phytochemicals, also providing dietary fiber and protein. Popularity of flax is steadily growing. Many flax products and ingredients for the animal and human food markets are under development. The scope of the project is to develop Bioflax, product based on flaxseed as a functional food and in the future to extend its function as preventive drug. Health-conscious consumers raise the demand for flax-enriched foods as they become more aware of the potential benefits of flax in reducing the risk of chronic diseases such as heart disease, stroke and cancer.Last minute info: Already on sale in Poland

6. Salvia officinalis and Escherichia coli transformation with Streptoccocus mutans genes – a new perspective in prophylaxis and treatment of dental caries

Dental caries is recognized as a social problem in the world. Streptoccocus mutans is strongly implicated as a causative organism of human and animal dental caries. Two major virulence factors of Streptococcus mutans have been implicated in the molecular pathogenesis of dental caries: antigen I/II and glucosyltransferase (GTF). Antigen I/II may function as an adhesin which mediates attachment of Streptococcus mutans to the tooth surface since it is essential for colonization in vivo and binds to salivary receptors in vitro. Glucosyltransferase enzyme (GTF) is considered as a second virulence factor characterizing Streptococcus mutans. Activity of GTF depends on synthesis water-insoluble glucans from sucrose. The glucans play an important role in dental plaque formation by facilitating the accumulation of bacteria on the teeth surfaces. Thus, antigen I/II and glucosyltransferase are the major candidates for a caries vaccine component. The aim of this project is to obtain vaccine against dental carries. Transformation of Salvia officinalis and Escherichia coli cells with Streptococcus

mutans genes (N-terminal and central fragments of antigen I/II, CAT and GLU regions of glucosyltransferase) may result in production of raw material for vaccinization.

Last minute info: Gene constructs for plant transformation are prepared. Application for GMO studies in progress. Estimated R&D cost U$ 400 000.

7. DNA bank of random Polish population

In last few years an important expansion of human DNA sampling and data collecting in order to exploit and study the genetic information collected was observed. The strategic importance of this activity for genetic research and its applications in biomedicine is very important. Human DNA, tissue or cell collections, Guthrie cards as well as databases, which are attached to such biological resources, are necessary for a wide range of purposes and these collections have been extensively exchanged for scientific purposes. However, the status of collections is not very well known and most laboratories that bank DNA have no written policies or agreements regarding this activity. Still, many DNA banks are concerned about how to obtain valid informed consent, safeguard the privacy of samples and data, and avoid potential misunderstandings with depositors. The rules that regulate access are still governed by practices that vary widely with the type of collection. The rules for exchange and sharing of information and material are not clear. The notion of return of benefits to the communities, which recognizes their contribution, is fairly recent.
Laboratory of Molecular Genetics was established in 1992. Since that time 20,000 DNA samples were deposited. DNA bank represents random sample of Polish population from whole country.

Last minute info: Ready for commercialization. Written consent for genetic analysis.

 

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