Why Should Prednisolone Suppress Hepatic Ischemia-Reperfusion Injury?

Hepatic ischemia-reperfusion injury is a serious complication but unavoidable problem in liver surgery including liver transplantation and hepatic resection. The most important consequence of this pathological process is multiple organ failure with a high mortality rate. Steroid therapy suppresses liver injury by a variety of mechanisms, including increased tissue blood flow and suppression of oxygen free radicals, arachidonic acid derivatives, lysosomal proteases (cathepsins) and cytokine production. However, the exact intracellular mechanisms of steroid action on hepatic ischemia-reperfusion injury remains unknown.

A research article published in the World Journal of Gastroenterology addresses this question. This research team was led by Prof. Meng Wang from Eastern Hepatobiliary Surgery Hospital of Shanghai.

The hepatic ischemia-reperfusion injury model was performed through clamped the left lateral and median lobes of rat liver (68%) for 60 minutes and followed by 120 minutes reperfusion. Prednisolone was administered at 1.0, 3.0, or 10 mg/kg at 30 min before ischemia. In addition to biochemical and microscopic analyses, activation of calpain mu was determined using specific antibodies against the intermediate (activated) form of calpain. Degradation of talin was also studied by Western blotting.

They found that in the control and prednisolone (1.0 mg/kg) groups, serum aspartate transaminase (AST) and alanine transaminase (ALT) level were elevated, and cell membrane bleb formation was observed after 120 min of reperfusion. Moreover, calpain mu activation and talin degradation were detected. Infusion of prednisolone at 3.0 or 10 mg/kg significantly suppressed serum AST and ALT, and prevented cell membrane bleb formation. At 10 mg/kg, prednisolone markedly suppressed calpain mu activation and talin degradation.

Their results indicate that prednisolone can suppress ischemia-reperfusion injury of the rat liver. Its cytoprotective effect is closely associated with the suppression of calpain mu activation and talin degradation.

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Reference: Wang M, Shen F, Shi LH, Xi T, Li XF, Chen X, Wu MC. Protective effect of prednisolone on ischemia-induced liver injury rats. World J Gastroenterol 2008;14(27):4332-4337
wjgnet/1007-9327/14/4332.asp

Correspondence to: Meng Wang, MD, Ph.D, Eastern Hepatobiliary Surgery Hospital, Second Military Medical University, 225, Changhai Road, Shanghai 200438, China.

About World Journal of Gastroenterology

World Journal of Gastroenterology (WJG), a leading international journal in gastroenterology and hepatology, has established a reputation for publishing first class research on esophageal cancer, gastric cancer, liver cancer, viral hepatitis, colorectal cancer, and H pylori infection and provides a forum for both clinicians and scientists. WJG has been indexed and abstracted in Current Contents/Clinical Medicine, Science Citation Index Expanded (also known as SciSearch) and Journal Citation Reports/Science Edition, Index Medicus, MEDLINE and PubMed, Chemical Abstracts, EMBASE/Excerpta Medica, Abstracts Journals, Nature Clinical Practice Gastroenterology and Hepatology, CAB Abstracts and Global Health. ISI JCR 2003-2000 IF: 3.318, 2.532, 1.445 and 0.993. WJG is a weekly journal published by WJG Press. The publication dates are the 7th, 14th, 21st, and 28th day of every month. WJG is supported by The National Natural Science Foundation of China, No. 30224801 and No. 30424812, and was founded with the name of China National Journal of New Gastroenterology on October 1, 1995, and renamed WJG on January 25, 1998.

About The WJG Press

The WJG Press mainly publishes World Journal of Gastroenterology.

Source: Lai-Fu Li

World Journal of Gastroenterology

Unigene Reports Positive Results On A Novel Peptide For Reducing Food Consumption in animals

Unigene Laboratories, Inc. will present data this week regarding a novel peptide that has been shown to reduce food consumption in animals. The poster, entitled “Reduction in Food Consumption and Weight in Dogs by Oral Delivery of a Novel Anorexigenic Peptide,” will be presented at the Keystone Symposium Conference on “Obesity: Novel Aspects of the Regulation of Body Weight” in Alberta, Canada.

The peptide, currently designated UGL269, is an analog of a natural peptide hormone, and was designed by Unigene scientists. It was tested against two other peptides in a placebo-controlled crossover study and demonstrated significant reductions in food intake and body weight when orally administered to dogs. The ability of UGL269 to reduce food intake was superior to that of other peptides reported to decrease food intake, including an analog of peptide PYY.

In the studies, oral administration of UGL269 resulted in a decrease in food intake by up to 50% and a significant decrease in body weight. Oral administration of an analog of peptide PYY resulted in a smaller reduction in food intake and minimal change in weight. Cessation of treatment with UGL269 resulted in a resumption of food intake to pre-dosing levels. Administration of placebo capsules had no effect on food intake.

“Therapies that modify feeding behavior and result in weight loss represent a significant unmet medical need,” commented Dr. Warren Levy, President and CEO of Unigene. “UGL269, which can be manufactured and orally delivered using our patented Secrapep® and Enteripep® technologies respectively, may offer a new, orally administered approach for weight loss through appetite reduction.”

About Unigene

Unigene Laboratories, Inc. is a biopharmaceutical company focusing on the oral and nasal delivery of large-market peptide drugs. Due to the size of the worldwide osteoporosis market, Unigene is targeting its initial efforts on developing calcitonin and PTH-based therapies. Fortical®, Unigene’s nasal calcitonin product for the treatment of postmenopausal osteoporosis, received FDA approval and was launched in August 2005. Unigene has licensed the U.S. rights for Fortical® to Upsher-Smith Laboratories, worldwide rights for its oral PTH technology to GlaxoSmithKline and worldwide rights for its calcitonin manufacturing technology to Novartis. Unigene’s patented oral delivery technology has successfully delivered, in preclinical and/or clinical trials, various peptides including calcitonin, PTH and insulin. Unigene’s patented manufacturing technology is designed to cost-effectively produce peptides in quantities sufficient to support their worldwide commercialization as oral or nasal therapeutics. For more information about Unigene, call (973) 265-1100 or visit www.unigene. For information about Fortical, visit fortical.

Safe Harbor statements under the Private Securities Litigation Reform Act of 1995: This press release contains forward-looking statements regarding us and our business, financial condition, results of operations and prospects. Such forward-looking statements include those which express plans, anticipation, intent, contingency, goals, targets or future development and/or otherwise are not statements of historical fact. We have based these forward-looking statements on our current expectations and projections about future events and they are subject to risks and uncertainties known and unknown which could cause actual results and developments to differ materially from those expressed or implied in such statements. These forward-looking statements include statements about the following: general economic and business conditions, our financial condition, competition, our dependence on other companies to commercialize, manufacture and sell products using our technologies, the ability of our products to gain market acceptance and increase market share, the uncertainty of results of animal and human testing, the risk of product liability and liability for human trials, our dependence on patents and other proprietary rights, dependence on key management officials, the availability and cost of capital, the availability of qualified personnel, changes in, or the failure to comply with, governmental regulations, the failure to obtain regulatory approvals for our products and other risk factors discussed in our Securities and Exchange Commission filings. Words such as “anticipates,” “expects,” “intends,” “plans,” “predicts,” “believes,” “seeks,” “estimates,” “may,” “will,” “should,” “would,” “potential,” “continue,” and variations of these words (or negatives of these words) or similar expressions, are intended to identify forward-looking statements. In addition, any statements that refer to expectations, projections, or other characterizations of future events or circumstances, including any underlying assumptions, are forward-looking statements. These forward-looking statements are not guarantees of future performance and are subject to certain risks, uncertainties, and assumptions that are difficult to predict. Therefore, our actual results could differ materially and adversely from those expressed in any forward-looking statements as a result of various risk factors.

Unigene Laboratories, Inc.

Options In Prostate Cancer Surgery: Perineal Prostatectomy In The Age Of Minimally Invasive Surgery

UroToday – In the age of minimally invasive surgery, laparoscopic and robotic assisted laparoscopic prostatectomy has been associated with a substantial increase in operative costs without a significant improvement in outcomes to date. As a result, there has been renewed interest in one of the original approaches to prostate cancer. In this presentation we compare the MUSC experience with perineal prostatectomy (RPP) to published results using laparoscopic and robot-assisted laparoscopic prostatectomy (LRP, RALP).

Our experience with over 300 consecutive perineal prostatectomies is presented and operative technique reviewed in detail. Issues such as hospital stay, intra-operative blood loss, length of catheterization, complications, both long and short term, and outcomes will be reviewed. Selective indications for a perineal prostatectomy will be defined and discussed.

The mean operative time for RPP was less than that for LRP or RALP. Estimated blood loss was equivalent for RPP and RALP and less than that for LRP. Length of catheterization in the RPP group was longer but was based on a care map and has been modified to 7 days without problems in the last 75 pts. Length of hospital stay was equivalent for all techniques and hospital costs were significantly less for RPP vs. the alternatives.

A detailed discussion of the current evidence and its quality concerning the various approaches will be undertaken and the audience should have a clear understanding of the current issues at the end of the presentation.

Presented by: Thomas Keane, MD, at the Masters in Urology Meeting – July 31, 2008 – August 2, 2008 – Elbow Beach Resort, Bermuda

UroToday – the only urology website with original content global urology key opinion leaders actively engaged in clinical practice.

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Copyright © 2008 – UroToday

Healthy People With Elevated Levels Of Uric Acid Are At Risk Of Developing Kidney Disease

Elevated uric acid levels in the blood indicate an increased risk of new-onset kidney disease, according to a study appearing in the December 2008 issue of the Journal of the American Society Nephrology (JASN). The results suggest that it may be appropriate to prescribe uric acid-lowering drugs, such as allopurinol and probenecid, to these otherwise healthy individuals.

High levels of uric acid in the blood, called hyperuricaemia, can be caused by a diet high in purines or by impaired excretion by the kidneys. Studies have linked hyperuricaemia to kidney failure, hypertension, and cardiovascular disease, but most of these studies have been conducted in people with metabolic abnormalities.

Rudolf Obermayr, MD, of Sozialmedizinisches Zentrum Ost der Stadt Wien, in Vienna, Austria, and his colleagues wished to determine the risks of elevated levels of uric acid in healthy individuals, particularly as they relate to kidney disease.

The investigators recruited 21,475 healthy volunteers from the general Viennese population, performing an initial examination and following up with three examinations over an average of seven years. Volunteers were categorized as having normal blood levels of uric acid, slightly elevated levels, or elevated levels. During follow-up examinations, the researchers assessed patients’ glomerular filtration rates, noting that low rates indicate decreased kidney function.

Dr. Obermayr’s team found that with increasing uric acid groups, glomerular filtration rates decreased. After eliminating all other potential risk factors that might contribute to kidney disease, the researchers determined that individuals in the slightly elevated uric acid group were 1.26 times as likely to develop kidney disease as those in the low uric acid group. The odds of developing kidney disease among volunteers in the elevated uric acid group were 1.63 times greater than that of individuals in the low uric acid group.

The risk of developing new-onset kidney disease was more evident in women than in men. It also was particularly high in individuals with hypertension. Dr. Obermayr noted that this finding may be important from a public health viewpoint, because the prevalence of prehypertension and hypertension in adults is approximately 60%, and the prevalence of hyperuricaemia is approximately 17%.

According to Dr. Obermayr, this study indicates that clinical trials assessing the potential of uric acid-lowering drugs for preventing kidney disease should be initiated. These agents, such as allopurinol and probenecid, are available worldwide and are safe and inexpensive.

This study was financially supported by the Medizinische Forschungsgesellschaft Donaustadt and by grants from the Austrian Science Fund (FWF P-18325) and the Austrian Academy of Science (OELZELT EST370/04).

The article, entitled “Elevated Uric Acid Levels Increase the Risk for New-Onset Kidney Disease,” appeared online at jasn.asnjournals on Wednesday, September 17, 2008, and in the December 2008 print issue of JASN.

ASN is a not-for-profit organization of 11,000 physicians and scientists dedicated to the study of nephrology and committed to providing a forum for the promulgation of information regarding the latest research and clinical findings on kidney disease. ASN publishes JASN, the Clinical Journal of the American Society of Nephrology (CJASN), and the Nephrology Self-Assessment Program (NephSAP). In January 2009, the Society will launch ASN Kidney News, a newsmagazine for nephrologists, scientists, allied health professionals, and staff.

Journal of the American Society Nephrology (JASN)

New Method Decodes Cell Movements, Accurately Predicts How Cells Will Divide

Researchers at Rensselaer Polytechnic Institute have discovered a new method for predicting – with up to 99 percent accuracy – the fate of stem cells.

Using advanced computer vision technology to detect subtle cell movements that are impossible to discern with the human eye, Professor Badri Roysam and his former student Andrew Cohen ’89 can successfully forecast how a stem cell will split and what key characteristics the daughter cells will exhibit.

By allowing the isolation of cells with specific capabilities, this discovery could one day lead to effective methods for growing stem cells on a large scale for therapeutic use.

“If you have many cells in a culture, they all look the same. But our new method senses all sorts of tiny differences in the shapes and movements of the cells, and uses these cues to predict what kind of cells it will divide into,” said Roysam, professor of electrical, computer, and systems engineering at Rensselaer. “We believe this method will be beneficial for one day taking cells from a patient, and then growing large amounts of the kind of cells that patient is in need of. This could enable many new and exciting types of medical treatments using stem cells.”

Results of the study, titled “Computational prediction of neural progenitor cell fates,” were published recently in the journal Nature Methods.

In order to achieve successful stem cell-based therapies, researchers require access to large amounts of specific cells. This has proven difficult, as there are currently no methods for controlling or manipulating the division of bulk quantities of cells. When stem cells or progenitor cells divide via mitosis, the resulting daughter cells can be self-renewing or terminal. A self-renewing cell will go on to split into two daughter cells, while a terminally differentiated cell is fated to be a specific, specialized cell type. Researchers want the ability to influence this division in order to produce large volumes of the correct type of cells.

Roysam and Cohen tracked the development of rat retinal progenitor cells cultured in their collaborator’s laboratory at McGill University. The computer system they developed took images of the cells every five minutes, and employed algorithmic information theoretic prediction (AITP) to observe the behavior of the cells, analyze the behavior, and discern whether each individual cell is fated to split into self-replicating or terminal daughter cells. This process occurs in real time, so researchers know the fate of cells before they actually divide.

The researchers predicted with 99 percent accuracy if the rat retinal progenitor cells would split into self-renewing or specialized cells, and predicted with 87 percent accuracy certain characteristics of the specialized cells.

“Our results suggest that stem cells display subtle dynamic patterns that can be sensed computationally to predict the outcome of their next division using AITP,” Roysam said. “In theory, AITP can be used to analyze nearly any type of cell, and could lead to advances in many different fields.”

Roysam said prototyping and development of the system leveraged the processing power of Rensselaer’s supercomputer, the Computational Center for Nanotechnology Innovations (CCNI).

Co-authors of the paper are Michel Cayouette and Francisco Gomes of the Cellular Neurobiology Research Unit at the Institut de Recherces Cliniques de Monteal; and Roysam’s former student Cohen, now an assistant professor of electrical engineering and computer science at the University of Wisconsin, Milwaukee.

This project was supported in part by the U.S. National Science Foundation Center for Subsurface Sensing and Imaging Systems, the Canadian Institutes of Health Research, and the Foundation Fighting Blindness-Canada.

Source:
Michael Mullaney
Rensselaer Polytechnic Institute

Electronic Cigarette Distributors Warned By FDA Regarding Unsubstantiated Claims

Warning letters were sent to five different electronic cigarette distributors by the FDA (Food and Drug Administration) for making unsubstantiated claims, as well as poor manufacturing practices. The FDA also informed the Electronic Cigarette Association that it means to regulate electronic cigarettes and related products “in a manner consistent with its mission of protecting the public health.”

A drug product is required to demonstrate its safety to the FDA, it also needs to prove it is effective for its intended use. Manufacturing methods have to be shown to be adequate so that the strength, quality and purity of the product is preserved, the Agency added.

In the letter to the Electronic Cigarette Association, the FDA wrote:

FDA invites electronic cigarette firms to work in cooperation with the agency toward the goal of assuring that electronic cigarettes sold in the United States are lawfully marketed.

Electronic cigarettes are subject to FDA regulation as drugs, the Agency has determined. It explains in its warning letters that under FDCA (Federal Food, Drug, and Cosmetic Act), if a company makes a claim to treat or mitigate a disease, such as nicotine addiction, it is required to prove its safety and effectiveness.

All five electronic cigarette distributing companies make this claim, but with no relevant evidence that their products help users give up smoking.

Warning letters were sent to the following companies:

E-CigaretteDirect LLC
Ruyan America Inc.
Gamucci America (Smokey Bayou Inc.)
E-Cig Technology Inc.
Johnson’s Creek Enterprises LLC.

Some firms received warning letters for other reasons too:

E-Cig Technology sells drugs in unapproved liquid forms, such as tadalafil, the active ingredient for the erectile dysfunction drug Cyalis, as well as rimonabant, a weight loss drug which has not been approved in the USA yet. Rimonabant is the active ingredient in the following brand named weight loss drugs, Acomplia, Bethin, Monaslim, Remonabent, Riobant, Slimona, Rimoslim, Zimulti, and Riomont. These liquid medications are designed to refill e-cigarette cartridges to that they may be inhaled through vapour.
Johnson Creek Enterprises, distributor of Smoke Juice, a liquid solution used to refill empty e-cigarette cartridges, is cited by the FDA for several faults in its manufacturing process, including not establishing quality control and testing procedures required under the FDCA.

What are e-cigarettes?
E-cigarettes, also known as personal vaporizers, or electronic cigarettes, are battery-powered devices that look like cigarettes which provide inhaled doses of nicotine in a vaporized solution – some may also provide non-nicotine vaporized solutions. They are sold as alternatives to smoked tobacco products, such as pipes, cigars and cigarettes.

E-cigarettes are said to provide a flavor and physical sensation similar to what a tobacco smoker experiences, but without the smoke or combustion (you don’t light it up).

Some e-cigarettes look just like cigarettes, while others look like ballpoint pens, and even pipes.

The majority of e-cigarettes are reusable and have replaceable and refillable parts. Some of them are disposable.

E-cigarettes in Europe – electronic cigarettes were brought to Europe in April 2006, and officially launched in Austria. Estimates place the number of regular e-cigarette users in Europe at over half-a-million.

E-cigarettes in the USA – classed as a drug by the FDA, and subject to market approval before sale under the FDCA. In January 2010 a federal judge overturned the FDA classification. However, an appeals court overruled the judge’s classification in March 2010. Several online e-cigarette companies have emerged online in America.

Sources: FDA, Wikipedia.

Doctors Use Patient’s Own Stem Cells To Grow Facial Bone In Groundbreaking Procedure

In a first-of-its kind procedure, physicians have used stem cells taken from the fat tissue of a 14-year-old boy and combined them with growth protein and donor tissue to grow viable cheek bones in the teen.

The new procedure dramatically improves the options surgeons have for repairing bone deficiencies caused by traumatic injuries such as those from car accidents or soldiers wounded in battle or by disease and genetic conditions, according to Jesse Taylor, M.D., a surgeon and researcher in the Division of Craniofacial and Pediatric Plastic Surgery at Cincinnati Children’s Hospital Medical Center. An estimated 7 million people in the United States have defects in bone continuity so severe that repair is difficult.

“We think this will benefit millions of people who, through traumatic injury or disease, have significant bone defects,” Dr. Taylor explained. “The current methods we have like borrowing bone from another part of the body, or implanting cadaver bone or something artificial are reasonable alternatives, but far less than perfect.”

Because the body rejects or absorbs implanted donor material, many reconstructive surgeries can have high failure rates. In procedures where bone is borrowed from one part of the body to replace another, the corrective surgery itself can be disfiguring to the person doctors are trying to help.

The new procedure avoids these problems because it uses the patient’s own cells, Dr. Taylor explained. His team developed the procedure based in part on scientific research conducted in pigs at Cincinnati Children’s. The operation is the first to blend and refine several techniques used or under study in surgical practice for repairing bone deficiencies.

The teenage recipient of the surgery, performed on May 28, has a rare genetic condition known as Treacher Collins syndrome, which includes underdeveloped or missing cheek bones. In this case the teenage patient, Brad Guilkey of Cincinnati, did not have developed zygomatic bones on either side of his face. The zygomatic bones form the prominence of the cheek and part of the outer rim of the eye socket.

The missing bones affected the active teenager’s appearance, but more importantly put his eyes at increased risk of injury, Dr. Taylor said. The bones are supposed to surround most of the lower and side areas of the eye sockets, with a portion protruding toward the ear at the cranial base.

“This bone is critical structurally and acts as a shock absorber for the face, protecting the eyes and other critical structures in the event of facial impact,” explained Dr. Taylor. “This young man is extremely active, he loves to play basketball and baseball, and growing new bone in this area of his craniofacial structure is critically important for him.”

Dr. Taylor said the procedure has been successful and, more than four months after the surgery, computer tomography (CT) scans show the teenager’s cheek bones have filled in normally with viable bone. The new bone structure enhances his appearance and improves protection for his eyes. Additional touchup surgery to the teenager’s eye lids is under consideration to address a slight downward slant, also characteristic of Treacher Collins syndrome.

During the day-long operation, surgeons used a section of donor bone to craft what essentially were mineral-based scaffolding implants (known as allografts), which also served as a growth guide for the new bone. Surgeons drilled holes in the allografts, which then were filled with mesenchymal stem cells taken from the patient’s abdominal fat. Also injected into the allografts was a growth protein called bone morphogenic protein-2 (BMP-2) that instructs the stem cells to become bone cells called osteoblasts.

One of nature’s roles for mesenchymal stem cells is to become cell types for a variety of different tissues in the body – including connective tissue and bone – giving the body a ready reserve of replacement cells as older cells die. In the surgery, and in the earlier lab experiments involving pigs, the doctors used BMP-2 to jumpstart nature’s normal process of transforming these malleable stem cells.

“We only need to use a fairly small amount of bone morphogenic protein to serve as a cue to tell the mesenchymal stem cells that they’re going to become bone,” explained Donna Jones, Ph.D., a researcher at Cincinnati Children’s and part of the scientific team that conducted experiments leading to the procedure. “The actual molecular mechanisms BMP-2 uses to do this are not well understood, but once we use BMP-2 to start the process, the body’s own biological processes take over and it produces its own BMP-2 to continue the transformation.”

Particularly critical to that process is wrapping the donor allograft bone in a thin membrane of tissue that coats bone surfaces called periosteum. The periosteum used in this surgery was taken from the patient’s thigh. Periosteum is important to the body’s normal production of BMP-2, and just as vital to providing a blood supply to nourish new bone formation.

Drs. Taylor, Jones and their fellow researchers are conducting ongoing studies into growing mandible bones in pigs. In a research paper being prepared for peer-review journal publication, they explain the use of the procedure to grow viable, dense bone in the animals and the duplication of results numerous times. The researchers worked with pigs because the porcine immune system is very similar to that of humans, making the animals a good model for simulating engineered bone growth in people.

Peer-review presentations of results from aspects of the study results have occurred at national re-constructive surgery conferences including the American Association of Plastic Surgeons and the Plastic Surgery Research Council and received with great enthusiasm, said Christopher Runyan, M.D., Ph.D., a member of the research team at Cincinnati Children’s.

The team also plans additional research projects to test the procedure’s ability to engineer bones of different lengths and sizes. Drs. Taylor and Jones said the technology may have the potential to grow almost any bone in the human body. As for Brad, now 15, and his mother, Christine, they’re just happy Brad can play sports and participate in other activities without having to worry about a lack of facial bone making him more susceptible to serious eye injury.

“Until we had the CT scans before surgery, we had no idea that Brad was missing the bones that protect his eyes, and that’s very dangerous,” said Christine. “I was nervous about the procedure, but we’re glad we did it and amazed with the results. The people at Cincinnati Children’s do a great job of explaining things to you and we have a lot of trust in the doctors and staff.”

About Cincinnati Children’s

Cincinnati Children’s Hospital Medical Center is one of 10 children’s hospitals in the United States to make the Honor Roll in U.S. News and World Reports 2009-10 America’s Best Children’s Hospitals issue. It is #1 ranked for digestive disorders and is also highly ranked for its expertise in respiratory diseases, cancer, neonatal care, heart care, neurosurgery, diabetes, orthopedics, kidney disorders and urology. One of the three largest children’s hospitals in the U.S., Cincinnati Children’s is affiliated with the University of Cincinnati College of Medicine and is one of the top two recipients of pediatric research grants from the National Institutes of Health.

President Barack Obama in June 2009 cited Cincinnati Children’s as an “island of excellence” in health care. For its achievements in transforming health care, Cincinnati Children’s is one of six U.S. hospitals since 2002 to be awarded the American Hospital Association-McKesson Quest for Quality Prize for leadership and innovation in quality, safety and commitment to patient care. The hospital is a national and international referral center for complex cases.

Source: Cincinnati Children’s Hospital Medical Center

Preventing Graft-Versus-Host Disease After Bone Marrow Transplant — Without Toxicity

Unless the donor is an identical twin, patients undergoing bone-marrow transplant (also known as hematopoietic stem cell transplant, or HSCT) must first receive powerful chemotherapy drugs to wipe out their immune system and prevent their bodies from rejecting the donated cells. Research from Children’s Hospital Boston and the Dana-Farber Cancer Institute has helped demonstrate that this punishing regimen increases the risk of graft-versus-host disease (GVHD), in which the donor’s cells mount an immune response against the patient. But the most recent findings also suggest that the risk for GVHD can be reduced by replacing a natural antibiotic protein, known as bactericidal/permeability increasing protein (BPI), which is depleted when patients undergo chemotherapy.

Now, a multicenter study is about to test this idea in HSCT patients, using a manufactured form of BPI known as rBPI21 (XOMA Ltd.) Unlike other treatments to prevent GVHD, BPI does not suppress the immune system and has shown virtually no toxicity.

Researchers Ofer Levy, MD, PhD, of Children’s Hospital Boston, and Eva Guinan, MD, of Children’s Hospital Boston and Dana-Farber Cancer Institute, presented their most recent findings and discussed the new clinical trial at the American Society of Hematology (ASH) Annual Meeting in Orlando, Fla. (abstract # 2856).

The new trial is the culmination of over five years of collaborative research by Levy and Guinan in human patients. “Many basic and translational studies, including our own, have provided a strong rationale for a trial of BPI in patients undergoing hematopoietic stem cell transplants,” says Levy. “Replenishing a natural host defense factor that is deficient due to chemotherapy makes theoretical and practical sense, and we hope that bringing our bench work to patients will reduce the complications they suffer.”

GVHD occurs when immune cells from donor attack the recipient, and can lead to multiple organ failure and death. It strikes some 30-60 percent of transplant patients, depending on how closely matched the donor is, and is kept in check only by eliminating otherwise useful donor immune cells or by using powerful immune-suppressing drugs.

Studies in mice had shown that the chemotherapy regimens used in HSCT not only wipe out white blood cells (with the intended effect of suppressing the immune system), but also damage the intestinal lining. This breach of the lining allows endotoxin, which is produced by bacteria living in the intestines, to enter the bloodstream. The endotoxin, in turn, provokes an inflammatory response that mobilizes donor immune cells, helping to trigger GVHD.

Levy, in Children’s Division of Infectious Diseases, had long been studying BPI, which naturally blocks and neutralizes endotoxin.(1) BPI is found in neutrophils, the very white blood cells that are virtually wiped out by pre-transplant chemotherapy. Studies in mice had shown that blocking endotoxin reduces the incidence of GVHD after chemotherapy and HSCT.(2)

Intrigued by these findings, Levy and Guinan began to study endotoxin and BPI in human patients undergoing HSCT with pre-transplant chemotherapy. In 2003 they showed, in a study of 57 children, that patients’ blood endotoxin levels rise markedly within a week of the transplant.(3) And now, in a study of 30 patient:donor pairs to be presented at the ASH meeting, they show that patients undergoing HSCT also have a sharp drop in BPI levels – just as their endotoxin levels are rising – and that BPI deficiency is associated with a greater likelihood of GVHD.

“BPI is markedly deficient – 100 to 1000-fold lower – in our transplant patients,” says Guinan, associate director of the Center for Clinical and Translational Research at Dana-Farber. “If we can replenish this host defense factor, we might be able to moderate the damaging effects of GVHD.”

The multicenter clinical trial, expected to begin within the next few months, will test rBPI21 (opebacan, NEUPREX® [Nasdaq: XOMA]). rBPI21 has been in phase I, II, and III human trials, with evidence of benefit in children and adolescents with serious meningococcal infections, but has not yet been approved by the Food and Drug Administration.

Levy and Guinan will first conduct a small safety trial, gradually increasing the amount of BPI given and the duration of treatment. If BPI appears safe, they will quickly mount a randomized, controlled trial in 30 to 40 patients who are undergoing HSCT for cancer or blood diseases. Children’s/Dana-Farber will be the lead center, with four to five additional pediatric and adult sites at prominent medical centers around the country.

“Our ultimate goal is to reduce the downstream complications of stem-cell transplant,” says Guinan. “BPI would make these transplants significantly less toxic.”

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The study presented at the ASH meeting was supported by XOMA and private donations to Guinan’s laboratory.

Founded in 1869 as a 20-bed hospital for children, Children’s Hospital Boston today is the nation’s leading pediatric medical center, the largest provider of health care to Massachusetts children, and the primary pediatric teaching hospital of Harvard Medical School. In addition to 347 pediatric and adolescent inpatient beds and comprehensive outpatient programs, Children’s houses the world’s largest research enterprise based at a pediatric medical center, where its discoveries benefit both children and adults. More than 500 scientists, including eight members of the National Academy of Sciences, 11 members of the Institute of Medicine and 10 members of the Howard Hughes Medical Institute comprise Children’s research community. For more information about the hospital visit: childrenshospital/newsroom.

Dana-Farber Cancer Institute (dana-farber/) is a principal teaching affiliate of the Harvard Medical School and is among the leading cancer research and care centers in the United States. It is a founding member of the Dana-Farber/Harvard Cancer Center (DF/HCC), designated a comprehensive cancer center by the National Cancer Institute.

References

(1) Levy O. A neutrophil-derived anti-infective molecule: bactericidal/permeability-increasing protein. Antimicrob Agents Chemother. 2000 Nov; 44(11):2925-31.

(2) Cooke KR, Gerbitz A, Crawford JM, Teshima T, Hill GR, Tesolin A, Rossignol DP, Ferrara JL. LPS antagonism reduces graft-versus-host disease and preserves graft-versus-leukemia activity after experimental bone marrow transplantation. J Clin Invest 2001 Jun; 107(12):1581-9.

(3) Levy O, Teixeira-Pinto A, White ML, Carroll SF, Lehmann L, Wypij D, Guinan E. Endotoxemia and elevation of lipopolysaccharide-binding protein after hematopoietic stem cell transplantation. Pediatr Infect Dis J. 2003 Nov; 22(11):978-81.

Contact: Anna Gonski

Children’s Hospital Boston

Mechanical ‘Artificial Hearts’ Can Return Heart To Normal And Remove Need For Heart Transplant

Mechanical ‘artificial hearts’ can be used to return severely failing hearts to their normal function, potentially removing the need for heart transplantation, according to new research.

The mechanical devices, known as Left Ventricular Assist Devices (LVADs), are currently used in patients with very severe heart failure whilst they await transplantation. The new study, published in the New England Journal of Medicine, shows that using an LVAD combined with certain drug therapies can shrink the enlarged heart and enable it to function normally once the LVAD is removed.

For the study, researchers from Imperial College London and the Royal Brompton and Harefield NHS Trust gave the full combination therapy to 15 severely ill patients. Of these 15, 11 recovered. Of these, 88 percent were free from recurrence of heart disease five years later. Their quality of life was measured as being at nearly normal.

Dr Emma Birks, from the Heart Science Centre at Imperial and the Royal Brompton and Harefield NHS Trust, and lead author of the study, said: “Donor heart transplant has for many years been the gold standard in the treatment of those with severe heart failure. It has proven greatly successful but is not without its shortcomings – particularly the shortage of donor hearts and the risk of organ rejection.

“This therapy has the potential to ease the pressure on the waiting list while also offering patients a better alternative to a donor heart – their own, healthy heart,” she added.

Professor Sir Magdi Yacoub, from the Heart Science Centre at Imperial and the Royal Brompton and Harefield NHS Trust, said: “We are impressed by the dramatic, sustained improvement in the condition of these severely ill patients and we believe that this is due to the additive effects of the particular combination therapy used. The improvement observed was far greater than what has been reported to date for any other therapy in patients with severe, but less advanced, forms of heart failure.

“The study also highlights the fact that ‘end stage’ heart failure can be reversed and that the heart has the capacity to regenerate itself. It therefore stimulates the search for other strategies and more therapeutic targets in this expanding field of regenerative therapy,” he added.

LVADs are currently mainly used in those patients awaiting heart transplant, whose heart failure is very severe. The researchers are hopeful that the technique used in this study could also be used to restore heart function amongst heart patients who are not awaiting transplants.

LVADs work by being connected to the left ventricle of the heart, either directly or by a tube. They remove oxygen rich blood from the left ventricle and take the blood to a mechanical pump. The mechanical pump then pumps the oxygen rich blood into another tube which is connected to the aorta. Once blood is in the aorta, it can be transported to the rest of the body.

Patients were treated with drugs which encourage reverse remodelling of the heart, prevent atrophy and prevent the heart from shrinking beyond its desirable size. The drugs used were lisinopril, carvedilol, spironolactone and losartan in the first stage of treatment and bisoprolol and clenbuterol in the second.

The next step for the researchers is a larger multi-centre trial named the Harefield Recovery Protocol (HARP) study, which is envisaged to start soon on both sides of the atlantic. The researchers are also continuing their molecular and cellular research and studying the mechanistic and therapeutic targets which have made the recovery observed in this study possible.

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The research was funded by Thoratec, the Royal Brompton and Harefield Charitable Trustees, the British Heart Foundation, and the Magdi Yacoub Institute.

Contact: Laura Gallagher

Imperial College London

Genetically Modified Stem Cells May Have Therapeutic Application To X-SCID

A study demonstrating therapeutic utilization of human stem cells, including potential application to X-linked Severe Combined Immunodeficiency (XSCID or “Bubble-Boy” disease) was presented at the 10th Annual Meeting of the American Society of Gene Therapy (ASGT) in Seattle.

X-linked Severe Combined Immunodeficiency (XSCID) is a rare genetic disease caused by defects in an essential immune hormone receptor called the common gamma chain (Оіc), found on the surface of lymphocytes and other immune blood cells. Patients with XCID have immune cells that lack this receptor and both fail to develop properly, as well as do not function normally to protect patients against infections.

A team of scientists led by Dr. Luigi Naldini from the San Raffaele Telethon Institute for Gene Therapy (HSR-TIGET), Milan and Sangamo BioSciences Inc., Richmond, CA, are developing a novel approach to genetically modify embryonic and tissue stem cells for therapeutic use. This approach can potentially be applied to many diseases, including X-SCID.

Using technology developed by Sangamo, the scientists showed gene correction or precise addition of a gene in human stem cells at unprecedented levels.

“This is a significant advance for the potential therapeutic use of stem cells,” says Dr. Angelo Lombardo, lead author of the study. “Stem cells are the body’s natural resource for regeneration and repair and the ability to efficiently insert a therapeutic gene in a predetermined location or to correct a mutated gene in a patient’s stem cells may enable us to provide a long term solution for many genetic diseases. The powerful combination of Sangamo’s ZFN™ technology and our highly efficient viral delivery platform was the key factor for obtaining unprecedented gene modification efficiency in these therapeutically important cells”.

Using this approach, an inherited mutation in a gene can be directly corrected or modified in a cell or a new copy of a gene can be added into the genome at a specific and predetermined site. This is a significant advance over traditional approaches that rely on the random insertion of a new copy of a gene to cells to restore the missing function of an inherited mutant gene. In fact, despite the successful outcome of gene therapy reported in many SCID patients, random insertion of genes into the cellular genome may in some circumstances lead to serious adverse effects due to altered expression of the therapeutic gene or its neighboring genes at the insertion site.

In contrast, Dr. Naldini’s study demonstrates that ZFN-mediated gene editing can be used to correct mutations in the IL-2R gamma gene, the defective gene in X-linked SCID, and to add a therapeutic gene to a pre-determined ‘safe-harbor’ site in the genome of both human hematopoietic progenitors and human ES cells with high efficiency. This ‘safe-harbor site’ was selected by the investigators for its capacity to allow efficient expression of the therapeutic gene and to tolerate an insertion event without adverse effects.

ZFN-mediated gene editing overcomes some major limitations of standard gene therapy approaches and potentially provides a unique means to treat disease with HSCs and ES cells in a safe and effective manner.

The scientists used novel designer proteins, named zinc finger DNA-binding protein nucleases (ZFN), that were developed at Sangamo BioSciences. ZFN can be engineered to specifically cut DNA at a chosen site.

Dr. Naldini’s team exploited the high infectious capacity of lentiviral vectors derived from HIV to express ZFN and provide the template DNA for gene correction in target cells. When they introduced ZFN into target cells with an appropriately designed donor DNA molecule that encodes the correct gene sequence, the DNA break was repaired in a natural process by the cell’s DNA-repair machinery leading to correction of the mutation or the precise insertion of a novel gene into the target site. Using the new delivery system, high rates of gene editing were obtained in a panel of human cells, including hematopoietic and embryonic stem cells.

The American Society of Gene Therapy is a professional non-profit medical and scientific organization dedicated to the understanding, development and application of gene and related cell and nucleic acid therapies and the promotion of professional and public education in the field. For more information, visit asgt.

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