During my early training in the laboratory of George E. Palade, I became convinced that the development or invention of new technologies provided the opportunity to make quantum leaps in scientific discovery. As a result of this belief I devoted substantial time to the invention of new techniques and strategies, which allowed my laboratory to make a number of seminal discoveries in the fields of protein sorting and trafficking, regulated secretion, regulation of gene expression, synthesis of full-length genes, prevention and early treatment of envelop viral infections and treatment of pulmonary diseases including Cystic Fibrosis. Further details on these inventions are shown below under nonpatented and patented categories:
TWO DIMENSIONAL GEL ELECTROPHORESIS
MICROSOMAL MEMBRANE TRANSLOCATION MODELS
PURIFIED RNase INHIBITOR
FLEX TECHNOLOGY FOR SYNTHESIS OF FULL-LENGTH GENE LIBRARIES
BICARBONATE TREATMENT FOR CYSTIC FIBROSIS
A METHOD FOR DIAGNOSING PANCREATITIS
TOPICAL PREVENTION AND EARLY TREATMENT OF VIRAL INFECTIONS
A method for preparing pancreatic tissue for structural (electron microscopy) and in-vitro functional studies in a manner that minimizes tissue damage and maximizes functional performance. This procedure is used beneficially with pancreatic tissue taken from guinea pig, rat or mouse. The procedure involves using a syringe and a needle to inject Krebs Ringer Bicarbonate (KRB) solution into the interstitial space thereby distending the tissue and revealing the smallest lobular structures perceived by the human eye. Then, using a pair of ophthalmic scissors, these lobular structures may be removed from the duct system and incubated in KRB under in vitro physiological conditions. Normally 5-10 lobules are incubated in a single Erlenmeyer flask at 37 degrees Celsius and in the presence of 95% oxygen and 5% carbon dioxide.
In experiments that conduct pulse-chase studies that rely on rapid transfer of lobules from one flask to another or rapid washing conditions, a piece of nylon mesh cut into a 2 cm circle, may be used to attach the lobules to a single mesh disk in each incubation flask. The connective tissue surrounding the lobules attach spontaneously to the nylon mesh. Following attachment of the lobules to the nylon mesh, which occurs immediately during shaking in a temperature-controlled water-filled incubator, a pair of forceps may be used to pick up the group of lobules for washing or transferring manipulations without introducing injury to the tissue itself.
Slide Presentation - Pancreatic Lobules
Ultilizing enzymatic digestion with collagenase and vigorous shearing forces, pancreatic lobules can be further reduced to pancreatic acini. However, there is an important functional difference between lobules and acini. In pancreatic lobules the acinar lumen is sequestered from the incubation medium allowing use of ductal inhibitors to manipulate the fluid, electrolyte and pH conditions of the acinar lumen. In pancreatic acini the conditions of the acinar lumen can be directly controlled by the conditions of the incubation medium. These functional differences allowed Dr. Freedman and myself to discover the role of bicarbonate secretion from duct cells in regulating the dissolution of secretory enzymes during exocytic release and the cellular uptake of exocytic membranes (secretory granule membranes inserted into the APM) by activated endocytosis at the APM.
Due to the fragile nature of tissue in the mouse pancreas, pancreatic acini obtained after treating with collagenase and shear forces, show relatively poor performance with high background levels of secretion. Thus, it is optimal to use pancreatic lobules when studying the mouse pancreas in vitro. Further, pancreatic lobules taken from mouse pancreas act more like acini in that the conditions of the acinar lumen appear to be directly controlled by the incubation medium. This observation suggests that small pancreatic ductile segments are severed during the tissue distension procedure.Back To Top
TWO DIMENSIONAL GEL ELECTROPHORESIS:
Two Dimensional Gel Electrophoresis separating proteins by charge and size - 2D separation of proteins in a slab gel utilizing isoelectric focusing in the first dimension and SDS gel electrophoresis in the second dimension allowed separation of proteins by charge and size, respectively. This procedure, developed independently by Dr. George Scheele and Dr. Patrick O'Farrell, allowed complex mixtures of proteins (up to 2000) to be separated by gel electrophoresis for the first time. Accordingly, it ushered in the era of "proteomics" twenty years before that term was coined. Invention of this technique allowed my laboratory to study the following biological processes for the first time:
- Kinetics of movement of the entire cadre of secretory enzymes through sequential intracellular compartments during the biogenesis and secretion of pancreatic enzymes.
- Kinetic of release of all secretory enzymes from the acinar cell during conditions of rest and secretagogue stimulation.
- Folding of nascent pancreatic enzymes following their sequestration into the RER lumen.
- Kinetics of activation of pancreatic proteases leading to the autoactivation of pancreatic juice, a pathological process believed to play a role in the pathogenesis of acute and chronic pancreatitis in humans.
- Gene and protein expression studies of families of pancreatic enzymes as a function of changes in ratios of nutritional substrates in the diet and as a function of secretagogue stimulation.
Slide Presentation - 2D gel ElectrophoresisBack To Top
MICROSOMAL MEMBRANE TRANSLOCATION MODELS:
Microsomal membrane models that allowed elucidation of N-terminal signal sequences responsible for the sequestration of secretory enzymes in the RER. The membrane model that proved to show the greatest success was the dog pancreas microsomes for the following reasons:
Compared to other organs, pancreatic microsomal membranes show the highest polypeptide chain translocation efficiencies.
Dog pancreas was superior to pancreas from other species because of low RNase levels (0.5 ug RNase per gm of tissue) and extraordinarily high RNase inhibitor levels (200:1 molar excess over RNase) in the cytosol.
Slide Presentation - Membrane Model for Protein Targeting & RER Sequestration in the Signal Peptide Hypothesis for Translocation of Presecretory Proteins into the RERBack To Top
PURIFIED RNase INHIBITOR:
A method by which purified RNase inhibitor is added to in-vitro translation systems to protect against contaminating RNase found in the translation components and the membrane fractions.
Slide Presentation - Purified RNase InhibitorBack To Top
FLEX TECHNOLOGY FOR SYNTHESIS OF FULL-LENGTH GENE LIBRARIES:
FLEX Technology for synthesizing full-length cDNA libraries - This patented technology utilizes poly-T priming followed by a reversible tailing procedure at the 5' end of single-strand cDNA to increase the representation of full-length double-stranded cDNAs from control rates of 10-20% to FLEX rates of 50-70%. This patent formed the basis for formation of Alphagene, Inc., a genomics company formed in 1993 and based in Boston , MA
- Scheele, G. and Fukuoka , S.-I. Synthesis of full-length, double-stranded DNA from a single-stranded linear DNA template. Patent no. 5,162,209, issued on 11-10-92.
- Scheele, G. Synthesis of full-length double-stranded DNA from a single-stranded linear DNA template. Patent no. 5,643,766, issued on 7-1-97.
PDF File - Flex Description
PDF File - Flex ProcedureBack To Top
BICARBONATE TREATMENT FOR CYSTIC FIBROSIS:
While conducting scientific research at Harvard Medical School , my laboratory determined the ultimate biochemical deficiency in Cystic Fibrosis and published the mechanism of this deficiency in the exocrine pancreas in a number of scientific publications. Detailed information on these discoveries appear in this website under "Scientist" and "Pancreatic Dysfunction in Cystic Fibrosis is Due to Impairments in Bicarbonate Secretion".
- Scheele, G. Treatment of pulmonary conditions associated with insufficient secretion of surfactant. Patent no. 5,863,563, issued on 1-26-99.
- This patent is held by Dr. George A. Scheele, the inventor.
Slide Presentation - Pancreatic Dysfunction in Cystic FibrosisBack To Top
TOPICAL PREVENTION AND EARLY TREATMENT OF VIRAL INFECTIONS :
Under the leadership of Dr. George A. Scheele, La Jolla Biosciences, LLC, has filed for three patents in the United States , with corresponding international filings, which claim the use of topical beta cyclodextrins for viricidal treatment of envelope viruses on topical surfaces of the body thereby preventing viral entry into the body. Drs. James E.K. Hildreth (Johns Hopkins Medical Institutions) and George A. Scheele (President, La Jolla Biosciences, LLC) are listed as co-inventors in the following three patent applications:
1. Prophylaxis, Systemic & Topical Treatments, Vaccines:
- U.S. Provisional Patent Application Serial No. 60,400,333. Compositions & Methods for Treating & Preventing Infection. Filed July 22, 2002 (6 applications)
- • U.S. Patent Application Serial No. 10/625,090, entitled Compositions & Methods for Treating & Preventing Infection. Filed July 22, 2003.
- PCT Application No. PCT/US03/22877 (WIPO), filed July 22, 2003.
SUMMARY OF CLAIMS:
- Broad protection covering use of cholesterol sequestering agents to prevent or treat infection by envelop viruses, including HIV, HTLV, Herpes, Hepatitis, Influenza & Pox viruses
- Oral & IV therapeutics
- Oral, IV, ID, IM & SubQ vaccines
- Topical treatment of viral diseases (Herpes and Influenza) on all epithelial surfaces.
- Protection for (myco-) bacterial, fungus, protozoan and non-envelope viruses (prevention of intracellular uptake)
- Extracorporeal treatment of envelope viruses.
2. Protection/Decontamination/Sanitation Formulations for the Health Care Industry:
- U.S. Provisional Patent Application Serial No. 60/402,318 - Compositions and Methods for Preventing Infection. Filed August 8, 2002 (3 applications)
- U.S. Patent Application Serial No. 10/637,793, entitled: Compositions & Methods for Treating & Preventing Infection. Filed on August 8, 2003
- PCT Application No. US03/24704 (WIPO), filed August 8, 2003.
SUMMARY OF CLAIMS:
- Broad protection covering use of cholesterol sequestering agents to reduce or prevent dermal transmission of envelope viruses, including HIV, HTLV, Herpes, Hepatitis, Influenza, SARS and Pox viruses.
- Protect Medical workers, blood handlers and soldiers.
- Gloves with an interior surface coated by BCD
- Treating environmental surfaces
- Uses of lotions, sprays and powders containing BCD.
Protection Technologies & Formulations for Blood Products:
- U.S. Provisional Patent Application Serial No. 60/412,399 - Compositions and Methods for Protecting Blood Products. Submission Date: September 20, 2002 (2 applications)
- U.S. Patent Application Serial No. 10/667,727, entitled: Compositions & Methods for Treating & Preventing Infection. Filed on September 22, 2003.
- PCT Application No. US03/29629 (WIPO), filed September 22, 2003.
SUMMARY OF CLAIMS:
- Addition of cholesterol-sequestering agents to whole blood, plasma, packed red blood cells, platelet concentrates, gamma globulin, purified plasma factors, albumin and factors VIII, IX and VIIA.
- Render viral-infected cells incapable of production of viable or functional viral particles
- Inhibit cells from uptake of envelop and non-envelope viruses
- Destroy free viral particles
- Filtration of 2-HP-BCD treated white blood cells
- Application of 2-HP-BCD to the blood circulation, cervical and vaginal canals to prevent envelope and non-envelope virus infection during childbirth to prevent maternal-to-fetal transmission of viruses.
Disclaimer :This product is not meant to diagnose, treat, cure, or prevent any disease by the use of pharmaceutical drugs. As a dietary supplement, with all natural ingredients, considered to be “generally regarded as safe” (GRAS), Factor4 is intended only to improve health and wellness in the organs and tissues throughout the body. Comments and notations, derived from testimonials and limited human studies, have not been evaluated by the FDA. All results stated in this white paper and on our website(s) are actual results from real customers or individuals included in our clinical trials. Individual results may vary depending on personal goals and use.