NIH Program Project Grant (P01 DK 43785): Ischemia-Reperfusion Injury
Founded: 1991 with 5-year grant for $4.7 million
Renewed: 1996 with 5-year grant ($6.4 million); renewed 2003 with 5 year grant ($8.0 million)
Full Time: 23
Part Time: 0
Budget: $19.1 million

Director: Dr. Neil Granger, Ph.D
Boyd Professor & Head, Department of Molecular and Cellular Physiology
LSU Health Sciences Center
1501 Kings Highway, P.O. Box 33932
Shreveport, LA 71130

Telephone: 318/675-6011 FAX: 318/675-6005 E-Mail: dgrang@lsuhsc.edu

Principal Investigators (Projects): D. Neil Granger, Ph.D.; Ronald Korthuis, Ph.D.; Steven Alexander, Ph.D.; Tak Yee Aw, Ph.D.; Matthew Grisham, Ph.D.

Principal Investigators (Cores): D. Neil Granger, Ph.D.;Tak Yee Aw, Ph.D.; Lynn Harrison, Ph.D.; Hugh V. Price, Ph.D.

Research Areas:

The inflammatory process is essential for survival, playing an important role in both health and disease. In recent years, there has been an explosion of knowledge concerning the immune system and its involvement in the inflammatory process. The identity, biological actions, and underlying molecular mechanisms of several classes of inflammatory mediators have been defined, and the contributions of different circulating, vascular and extravascular cell populations to the inflammatory response are now well appreciated. A major conceptual outgrowth of the intensive effort to define the inflammatory process is the realization that vascular endothelial cells play a pivotal role in the coordination and regulation of immune responses that are elicited in inflamed tissue. This critical role for the endothelial cell reflects both its ability to respond to inflammatory mediators (eg, cytokines, oxidants) as well as its ability, when activated by these mediators, to orchestrate the recruitment of circulating leukocytes into inflamed tissue by creating a favorable environment for their trafficking, capture, and extravasation. Other consequences of the endothelial cell activation that accompanies an inflammatory response include an impaired ability of arterioles to regulate blood flow, increased fluid filtration across capillaries, and enhanced plasma protein extravasation in postcapillary venules. Collectively, these microvascular responses sustain the leukocyte infiltration, hyperemia and interstitial edema that are considered to be the cardinal features of inflammation.

Inflammatory processes have been implicated in a variety of human diseases, including the more traditionally defined chronic inflammatory diseases, such as arthritis and inflammatory bowel disease, as well as systemic cardiovascular diseases (atherosclerosis) and disorders (circulatory shock) for which the involvement of an inflammatory response has only recently been recognized. The diversity of the list of human diseases that involve inflammation is matched by an equally diverse list of models that are used to mimic the inflammatory process in experimental animals. Chemical irritants, immune activators, cytokines, and other exogenous agents have been widely used to elicit acute and/or chronic inflammatory responses in animal models. Other models have exploited the intrinsic ability of tissues to generate inflammatory mediators following a period of oxidative, ischemic or hypoxic stress, thereby negating the need to expose tissues to exogenous agents.

An experimental insult that has been widely employed to elicit and study the mechanisms underlying the microvascular and tissue responses to inflammation is ischemia and reperfusion (I/R). While tissue hypoxia and acidosis are generally regarded as the major factors that mediate the pathological alterations associated with ischemia per se, an inflammatory process has been invoked to explain the vascular dysfunction and tissue injury produced by I/R. The intensity of this inflammatory reaction in some postischemic tissues (eg, intestine) can be so severe that the reperfusion-related response is also manifested in distant organs. The remote effects of I/R are most frequently observed in the lung, liver, and cardiovascular system, and can result in the development of the systemic inflammatory response syndrome (SIRS) and multiple organ dysfunction syndrome (MODS). Although I/R induced inflammatory responses and the accompanying microvascular dysfunction have been described in most organ systems, the splanchnic circulation appears to be particularly vulnerable to the deleterious effects of such an insult. Hence, it is not surprising that the literature is replete with reports that implicate I/R in the pathogenesis of a variety of digestive diseases (eg, duodenal ulcers, inflammatory bowel disease) and conditions (liver transplantation) that are characterized by profound changes in tissue perfusion and/or an intense inflammatory response. While it remains unclear whether splanchnic I/R injury is amenable to therapeutic intervention, this physiologically relevant stimulus continues to offer unique insights into the intrinsic ability of tissues to initiate and sustain an inflammatory response. Hence, a better understanding of the processes involved in I/R injury may therefore ultimately provide us with information about how to protect tissues against the deleterious consequences of ischemic episodes as well as acute inflammatory events not associated with an ischemic insult. The extensive experimental characterization of the contribution of different adhesion molecules to leukocyte recruitment as well as the role of reactive oxygen metabolites in mediating the increased vascular permeability in intestinal I/R are two examples of the novel insights that have emerged from this model of inflammation. Over the past 10 years, the investigators participating in this Program Project Grant (PPG) have contributed significantly to the existing body of knowledge in the area of I/R injury and have taken a leadership role in proposing and testing novel concepts that relate to the molecular mechanisms that underlie the inflammatory and microvascular responses elicited by I/R.

Reactive metabolites of both oxygen (eg, superoxide) and nitrogen (eg, nitric oxide) have been implicated in the pathogenesis of most forms of inflammation. These reactive nitrogen oxide species (RNOS) appear to contribute to the inflammatory process by: 1) serving as signalling molecules, 2) activating nuclear transcription factors for both pro- and anti-inflammatory proteins, and 3) mediating cell necrosis and apoptosis. While nitric oxide (NO) and superoxide per se are often ascribed anti- and pro-inflammatory roles, respectively, the products of their chemical interaction (RNOS) can yield either phenotype (anti- or pro-inflammatory), depending on whether there is net oxidation or nitrosation of specific molecular targets that regulate the inflammatory response. These observations have led to the concept that a critical determinant of the inflammatory (pro- or anti-) phenotype assumed by tissues is the existing balance between reactive oxygen species (ROS) and NO. In no inflammatory state has this NO/ROS balance concept received more attention than in I/R, wherein an imbalance between NO and ROS is believed to produce the pro-inflammatory phenotype that is characteristic of postischemic tissues. Despite the existing body of evidence that supports a role for NO and ROS in the pathogenesis of I/R injury, no effort has been made to systematically assess the relative importance of ROS and NO in I/R-induced inflammatory responses and to determine the biochemical and molecular basis for this NO/ROS-mediated response.

The overall objectives of this PPG are: 1) to define the biochemical and cellular responses that ultimately lead to the organ dysfunction that occurs following reperfusion of ischemic tissues, and 2) to elucidate the molecular mechanisms underlying these responses. This PPG includes a spectrum of projects that will focus on the critical role of endothelial cells and inflammatory cells in the pathogenesis of I/R injury in intestine and liver. A multidisciplinary approach will be used to elucidate the biochemical, molecular, structural, and physiological responses of the microvasculature to ischemia and reperfusion. The work outlined in this PPG is a natural outgrowth of the research interests and ongoing collaborative efforts of eleven investigators with active programs in the areas of endothelial cell biology, nitric oxide/oxygen radical biochemistry, inflammation, microcirculation, and gastroenteric biology. The 5 highly focused and strongly inter-related projects in this Program rely on 4 core units (1 administrative & 3 scientific) which provide state-of-the art technical support that will help to ensure that the scientific objectives of each project are met.

Areas of Expertise:

This Program formalized and extended existing collaborative research efforts among 11 highly productive scientists at LSUHSC-S with active, well-funded programs in the areas of endothelial cell biology, inflammation, microcirculation and gastroenteric biology. The funding of this PPG allows investigators with diverse backgrounds and expertise to focus on multiple aspects of a single problem, the pathophysiology of intestinal I/R, thereby facilitating the formulation and testing of new hypotheses relative to this question. This coordinated effort involves a multidisciplinary approach aims at elucidating the mechanisms responsible for intestinal ischemia/reperfusion injury at the molecular, cellular, single microvessel and organ levels. The ultimate goal of the PPG is to obtain the fundamental knowledge that is needed to diagnose and treat ischemic disorders of the bowel.

Special Capabilities and Facilities:

Of the over 400 PPGs funded by the National Institutes of Health, this is the only Program which focuses on the mechanisms of ischemia/reperfusion injury. Another unique aspect of this Program relates to the multidisciplinary, interdepartmental approach that brings to bear the scientific expertise of a group of investigators with diverse backgrounds, research techniques and experience to focus on a single problem. The Program also supports 4 core facilities including the Administrative, Cell Culture and Imaging, Biochemistry and Molecular Biology and an Induced Mutant Mouse Core that function to provide support for, as well as to integrate and standardize the activities of the Program.

Research Equipment:

A wide variety of state-of-the-art equipment is available for studying the cellular and molecular mechanisms of ischemic disorders. Of particular note are the Confocal Imaging Microscope and Ratio Fluorescence Spectrometer facilities.

Keywords:

Ischemia/reperfusion
Endothelial cells
Cardiovascular Disease
Cell culture
Molecular biology

LIIPAT - Section for Vascular Endothelial Cells (Haraldsen Group)
Vascular endothelial cells line the inside of blood vessels. Once activated, they follow different programs, either getting sticky and recruiting leukocytes to sites of inflammation, or leaving the vessels themselves to build new blood levels. We focus on molecular mechanisms of endothelial cells differentiation with special emphasis on properties that enable leukocyte recruitment in immunity and inflammation, as well as those that enable vascular morphogenesis.


The current model of leukocyte adhesion to the vascular wall is viewed as a multi-step adhesion pathway. Chemokines presented at the endothelial cell surface enable the transition of leukocyte behaviour from rolling to firm adhesion, and are the focus of interest because they attract different subclasses of leukocytes. At sites of acute injury, leukocyte recruitment can occur rapidly, partly explained by the fact that endothelial cells store and rapidly release preformed chemokines to the vascular surface upon exposure to histamine or thrombin. We are currently studying the sorting profile of different chemokines in endothelial cells and the molecular mechanisms of their sorting.

To understand the biology of endothelial cells in different vascular segments and organs we compare the gene expression profile of endothelial cells of different tissue origins, having identified the first nuclear factor of a specialized subtype of vascular endothelial cells (NF-HEV), the characterization of which is ongoing.

In a different approach to understanding of vascular differentiation, we also developed a model for the adoptive transfer of endothelial cells. This technique is based on transferring single human endothelial cells to subcutaneous gels in immunodeficient mice in which they assemble to complex, functional vessels that make xenogeneic contact with the murine vascular system. Importantly, phenotypic properties that are lost in vitro are partly regained upon transfer, and by means of local, osmotic pump-delivery of soluble mediators to these gels, our method is a powerful tool for exploring the regulation of endothelial differentiation through direct experimental manipulation in vivo .







--------------------------------------------------------------------------------
Editors: Editorial board, patologi-post@labmed.uio.no
Document created: 06.11.2001, modified: 10.02.2005, certified: 06.11.2001
Get in touch with the University of Oslo

Dear Vicc,

I have been back on my COQ10 for the last couple weeks.

http://ajpheart.physiology.org/cgi/c...ct/278/4/H1084

I turly am in 100% agreement that the Chambers need people very compatent. As well I think we need complete Pulmonary testing. Each one of us are oxygen levels could be different. I feel a T. Vascular surgeon should also be consulted.

I have heard of a couple people going into remission with this HBOT treatment. But why only a couple?

Not only have I heard the nerves and blood work together but I have also been told the lung and heart work together as well.

As far as Drexel University, I have seened Maleki MD. He is very knowledgeable and has a heart truly in the right place. He did not promise me remission, but pain control. He also uses Ketamine. I was in his office for about a hour he charged me barely anything.

Vicc, have you talked to any T. Vascular surgeons on this therapy? Because some that I spoke with are very aware that RSD has a vascular component.

We are all bonded by this RSD stuff. I know in your heart you want to see all of us cured. I thank you for that from the bottom of my heart. Big Hugs, Roz

Hi Everyone,

Just another article. Please read the whole thing. What I have copied is about 1/3 down.

http://www.emedicine.com/neuro/topic602.htm

IL-6 in clinical stroke

Several clinical studies in patients with stroke have found an association between CSF and plasma IL-6 levels and infarct size and neurologic outcome. Some of these studies have yielded conflicting results.

Fassbender et al (1994) found that plasma levels of IL-6 showed a significant increase within the first few hours following stroke onset, peaking at 10 hours. This increased level of IL-6 significantly correlated with the volume of the brain lesion and was associated with a poor functional neurologic outcome. In contrast, serum levels of IL-1b and TNFa did not increase.

Different results were obtained by Tarkowski et al (1995). In this study, CSF levels of IL-6 were elevated significantly by day 2 and were correlated with the volume of infarct observed; serum levels of IL-6 did not correlate with the size of brain lesion. This study did not find any measurable levels of plasma IL-1b. Finally, Kim et al (1996) measured IL-6 levels in 29 patients with acute stroke. They found the level of IL-6 to be highest (49±16 pg/mL) at 24 hours after onset, remaining high (14±4 pg/mL) for at least 7 days.

The role of cytokines in clinical stroke also has been investigated. Plasma levels in vivo of IL-6 and the naturally occurring IL-1ra were measured using ELISA in 50 patients with acute stroke (4±2 days after onset) and in 20 age-matched healthy controls.

Levels of both IL-6 and IL-1ra were significantly higher in the stroke population (4.6±4.2 pg/mL and 354±270 pg/mL, respectively) than in controls (1.0±0.9 pg/mL and 139±113 pg/mL). The levels of both cytokines were significantly higher in patients who had infarcts measuring more than 3 cm on CT scans. We also found that the level of IL-6 is elevated not only in the acute stroke period, but it remains elevated for up to a year (compared to controls) upon longitudinal follow-up (Coull, 1993).

The acute IL-6 elevation and its ability to predict neurologic recovery from stroke has been investigated. In this study, plasma levels of IL-6, fibrinogen, and albumin along with white blood cell (WBC) count were measured within 4±2 days of onset in 131 patients with stroke. Standard clinical predictors of outcome also were obtained, including neurologic assessments and head CT infarct sizes.

Peak levels of IL-6 were correlated significantly (r=0.18) with 6-month outcome as assessed by the Glasgow Outcome Scale. Taken together, the acute-phase response variables strongly predicted 6-month recovery (R2=0.31) and were nearly as strong a predictor as the standard clinical predictors (R2=0.38). Recent clinical studies have confirmed these findings and found that IL-6 values strongly correlated with C reactive protein (Smith et al, 2004). Acalovschi et al (2003) recently confirmed the robust elevation of serum IL-6 post clinical stroke but found that the soluble receptors (antagonists) for IL-6 were downregulated. These clinical results suggest that the acute-phase response, in particular IL-6, is strongly associated with acute ischemic stroke; the acute-phase response also appears to be correlated with initial infarct size and degree of long-term recovery.

Cytokines - Conclusions

Taken together, the aforementioned observations strongly suggest that the

Roz -

A friend of mine just sent me the full text of "Dietary coenzyme Q10 supplement renders swine hearts resistant to ischemia-reperfusion injury." At the bottom of the first page there's the following notation:

The costs of publication of this article were defrayed in part by the payment of page charges. The article must therefore be hereby
marked ‘‘advertisement’’ in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.

Do you have any idea what that means? Is it something other than a peer reviewed article, notwithstanding the fact that it was published by the American Physiological Society?

Mike


P.S. If anyone wants a copy of this by email, drop me a pm by no later than 9:00 a.m. Pacific time on December 28th with your email address and I'll be glad to send it on. Otherwise it will have to wait until I get back from a 12 -day meditation retreat.

Hi Everyone,
Here is the Full Article on "Dietary coenzyme Q10 supplement renders swine hearts resistant to ischemia-reperfusion injury." If anyone has any opinions on this I would appreciate it. Big Hugs, Roz

http://ajpheart.physiology.org/cgi/c...ll/278/4/H1084

Looks like if they catch it quickly, and start the treatment there is a good chance of not having any damage to the heart. This would help the thousands of people that suffer heart attacks, and strokes every year.

Im not sure how it would help people that already have the damage, or people suffering with RSD. Unless it would somehow help the damage from spread?

Great news though, and I hope it gets to helping people soon!

Hi Roz,

It would have been nice to see these articles posted while I was still trying to teach everyone what ischemia-reperfusion injury (IRI)
is, and using the words found in your posts; they would have shown that I at least knew what words to use and how to spell them.

It is impossilbe to understand RSD without understanding IRI, but it is possible to understand explanations of what IRI does, and to see the connection between that and what takes place in RSD; all you need do is trust that the person explaining IRI is scrupulously accurate.

In order to show that I'm being scrupulously accurate, all future major posts I make will cite research that anyone can read, and offer copies of that research to anyone who requests a copy. Everyone is also free to search IRI on the Internet and reach their own conclusions.

About the pig heart research: IRI was discovered after open-heart surgeries led to high patient mortality following apparently successful surgeries. Finding an answer was critical, so lots of researchers jumped onto the problem, and found the answer.

Once they found out what was killing patients, the search focused on how to prevent IRI. Researchers found that simply applying vitamin E to the heart-wound itself resulted in a dramatic reduction of post-surgical IRI. Vitamin E is an antioxidant and this has been confirmed many times as the mechanism that prevents IRI.

Since then, researchers began looking in two directions:

They began to investigate whether IRI occured in other internal organs following surgeries. They found that it does in nearly every organ in the body. More recently, they have found that IRI is found in skeletal muscle; so far they have only investigated this disorder as something that follows a surgical intervention (transplant, surgical flaps, etc).

They have not looked into the possibility that non-surgical trauma (blunt force trauma) can cause IRI because they have no reason to look; they have never heard of an instance in which this sort of trauma caused IRI. A vascular surgeon did begin to look at the possibility that RSD is actually IRI, but he is underfunded and his preliminary findings ignored by the RSD experts. I read what he wrote and began investigating IRI. I became convinced that this is an IRI, and since then have been writing about what I learned.

The second direction IRI research took was to find more effective ways to prevent IRI; vitamin E works much of the time, but it can't be applied until the end of the surgery. The pig heart study is focused on finding a medical intervention that begins this preventive process earlier. I don't believe that preventive measures against IRI will be of much use in RSD, because this disease is so rare there is no point in trying to prevent it; it won't happen to the vast majority of the population anyway.

Mike, I had planned to let our debate end with your reply; I said what I wanted to say, you replied, let everyone decide for themselves. I would have done exactly that except you misrepresented my words twice, and while others may not have noticed, I did.

Your misrepresentations did not involve questioning my accuracy in reporting medical facts, but they did distort what I said about doc S.

You wrote: To suggest without any evidence that Dr. Schwartzman was/is receiving kickbacks from these hospitalizations is, in my opinion, not only outrageous but actionable.

I said that S is a fraud who is making enormous profits from a questionable procedure. I did not suggest that he is getting kickbacks. I don't know who gets what, but I know that $25,000.00 for a relatively inexpensive procedure is too damn much money.

You wrote: On the other hand, if your point is that no effective work can be done so long as one man or group of men lives, then I don't know what to say.

I wrote that doc S and his cohorts were barriers to understanding the real nature and etiology of RSD, that he's old and should die soon and this will remove a significant barrier. I also wrote that I didn't plan to sit idly by waiting for his death; that I felt the barrier he presents is not impossible to overcome, and if the truth emerges despite him, he would be irrelevant.

But now that I'm here, there are a couple of other comments I'd like to make. The enterepenurial doctors are a straw man; they have nothing to do with the main thrust of my post, which was that there is no evidence of traumatic nerve damage in RSD (CRPS-I or CRPS-II). This forum does not, and should not, limit the introduction of extraneous ideas, but what does this discussion have to do with what I was talking about?

I would like to see you challenge the major points that I raised here; present evidence showing that RSD is the result of nerve damage. I don't know why you offered Oaklander or Albrecht, since neither of them claimed their findings showed a neurological etiology for this disease. If you can't do that, perhaps you could argue that I was wrong in saying the SNS view has been discredited; or maybe refute my reasoning in rejecting central sensitization as applicable to RSD.

I could go on, but I've previously presented research showing why sympathetic blocks affect normal -- not abnormal -- SNS functioning, and we all know that blocks don't substantially affect the future course of RSD; it gets worse with or without them.

Hoping for a more focused discussion related to the actual material found in my original post...Vic

i am new to this forum, and in fact gave up forums and support groups years ago, for lack of new knowledge .... i am impressed with the conversation on this thread. i was an icu nurse before rsd, and am sad to say i had never heard of it. since rsd, i have read it all ... and it all says nothing. this is the first lively, and well researched conversation i have seen in all my almost 11 years of rsd ... thanks for the banter and the work done here.
i look forward to continued reading and you got me going, i'm alreadt looking up iri.

Hi,
Co Q 10 is not new to me. I have taken it for years. Prior to that I did a lot of research and I have done a lot of CEU's for renewal of my nursing license on supplements. I continue to study it / them esp since I read a survey that doctors (mostly cardiologist) who answered the survey question
"What would You take if you could only take ONE daily supplement?"
Almost all said Ubiquinone (sp?) or Co Q10.

My husband's cardiologist recommended it as well as pharmaceutical grade fish oil. Both of these are expensive because you need to take pure fish oil w/o contaminates and the Co Q10 needs to be of the highly absorbable type. Bouth are touted as useful in neuroligical diseases. I've done my own research over the years and never saw Co Q fail, helping my friend to the point during congestive heart failure to reverse it and he was able to climb stairs. It is said it will provide oxygen to tissues. I know it stops the color changes on days I remember to take it, esp every 12 hrs.
I can't speak of the man whose research I follow because he sells the stuff.
I've never told anyone about it except suffering friends, until right here, right now. As we age, levels in our blood decline. To me that is one answer to why our hearts "sometimes" may weaken as we age even tho one may not have had a heart attack. “It is unthinkable for me to practice good cardiology without the help of Coenzyme Q10. And, for the thousands of people with cardiac conditions so severe that they need a heart transplant, CoQ10 may be a suitable alternative that not only enhances the quality of life, but extends survival as well.”
S.S., MD, FACC, Cardiologist & Author, The Coenzyme Q10 Phenomenon
“Energy is life, and CoQ is a crucial component of the energy cycle and therefore of life itself”
Emile Bliznakov, MD Researcher and Gerald Hunt, co-authors, The Miracle Nutrient: Coenzyme Q10
If you'd like more info please PM me
Hope