Warren symposium follows legacy of geneticist giant

If we want to understand how the brain creates memories, and how genetic disorders distort the brain’s machinery, then the fragile X gene is an ideal place to start. That’s why the Stephen T. Warren Memorial Symposium, taking place November 28-29 at Emory, will be a significant event for those interested in neuroscience and genetics. Stephen T. Warren, 1953-2021 Warren, the founding chair of Emory’s Department of Human Genetics, led an international team that discovered Read more

Mutations in V-ATPase proton pump implicated in epilepsy syndrome

Why and how disrupting V-ATPase function leads to epilepsy, researchers are just starting to figure Read more

Tracing the start of COVID-19 in GA

At a time when COVID-19 appears to be receding in much of Georgia, it’s worth revisiting the start of the pandemic in early 2020. Emory virologist Anne Piantadosi and colleagues have a paper in Viral Evolution on the earliest SARS-CoV-2 genetic sequences detected in Georgia. Analyzing relationships between those virus sequences and samples from other states and countries can give us an idea about where the first COVID-19 infections in Georgia came from. We can draw Read more
•••

News

Making “death receptor” anticancer drugs live up to their name

Cancer cells have an array of built-in self-destruct buttons called death receptors. A drug that targets death receptors sounds like a promising concept, and death receptor-targeting drugs have been under development by several biotech companies. Unfortunately, so far results in clinical trials have been disappointing, because cancer cells appear to develop resistance pathways.

Death receptor-targeting drugs under development include: drozitumab, mapatumumab, lexatumumab, AMG655, dulanermin.

Winship Cancer Institute researcher Shi-Yong Sun, PhD and colleagues have a paper in Journal of Biological Chemistry that may help pick the tumors that are most likely to be vulnerable to death receptor-targeting drugs. This could help clinical researchers identify potential successes ahead of time and maximize chances of a good response for patients.

Postdoctoral fellow Youtake Oh is the first author. Winship deputy director Fadlo Khuri, MD and Taofeek Owonikoko, MD, PhD, co-chair of Winship’s clinical and translational research committee, are co-authors. Khuri’s 2010 presentation on death receptor drugs and lung cancer is available here (PDF).

Sun’s team shows that mutations in the cancer-driving genes Ras and B-Raf both induce cancer cells to make more of one of the death receptors (death receptor 5). In addition, they show that cancer cells with mutations in Ras or B-Raf tend to be more vulnerable to drugs that target death receptor 5.

Shi-Yong Sun, PhD

These mutations are known to be more common in some types of cancer. For example, roughly half of melanomas have mutations in B-Raf. Vemurafenib, a drug that inhibits mutated B-Raf, was approved in August 2011 for the treatment of melanoma. K-ras mutations are similarly abundant in lung cancer.

The selection and targeting of tumors via their specific mutations is a growing trend. Sun says lung, colon and pancreatic cancer are all cancer types where Ras and Raf mutations are common enough to become useful biomarkers. In lung cancer, Sun’s team’s results could be especially welcome news because, as a 2009 review concluded:

Recent studies indicate that patients with mutant KRAS tumors fail to benefit from adjuvant chemotherapy, and their disease does not respond to EGFR inhibitors. There is a dire need for therapies specifically for patients with KRAS mutant NSCLC.

 

Posted on January 10, 2012 by Quinn Eastman in Cancer Leave a comment

New molecular target in dystonia

Emory researchers led by pharmacologist Ellen Hess have identified a new molecular target in dystonia. Their findings, recently published in the Journal of Pharmacology and Experimental Therapeutics, could help doctors find drugs for treating the movement disorder.

Ellen Hess, PhD

Dystonia gives sufferers involuntary muscle contractions that cause rigid, twisting movements and abnormal postures. It is the third most common movement disorder, after tremor and Parkinson’s disease. Neurologists can sometimes use drugs to address the symptoms of dystonia but there is no cure.

A 2008 review by Hess (PDF) concludes that compared with other neurological disorders, “our understanding of the biology and potential treatments for dystonia is in its infancy.” Still, scientists have known for a while that the cerebellum, a region of the brain that regulates movement, is involved.

“We focused on the cerebellum because studies in patients with dystonia often show that this part of the brain is more active, when examined by MRI,” Hess says. “The abnormal overactivity of the cerebellum is seen in patients with all different types of dystonia, so it seems to be a common hotspot. Our goal was to understand what might be causing the overactivity in mice because if we can stop the overactivity, we might be able to stop the dystonia.”

Hess and her colleagues discovered that drugs that stimulate AMPA receptors induce dystonia when introduced into the mouse cerebellum. Their results suggest that drugs that act in reverse, blocking AMPA receptors, could be used to treat dystonia.

Postdoctoral fellow Xueliang Fan is the first author of the paper. Emory neurologist H.A. Jinnah, director of a NIH-supported network of clinical research sites focusing on dystonia, is a co-author.

AMPA receptors are a subset of glutamate receptors, a large group of “receiver dishes” for excitatory signals in the brain. Fan performed a variety of experiments to show that AMPA receptor activity plays a specific role in generating dystonia. For example, drugs that affect other types of glutamate receptors did not induce dystonia. AMPA receptor blockers can also reduce dystonia in a genetic model, the “tottering” mouse.

Although pharmaceutical companies have already been testing AMPA receptor blockers as potential antiseizure drugs, caution is in order. AMPA receptor stimulators/ enhancers (or “ampakines”) have been identified as potential enhancers of learning and memory, so AMPA receptor blockers may interfere with those processes.

“Our results suggest that reducing AMPA receptor activity could help alleviate dystonia but we still have a lot of work to do before we know whether blocking AMPA receptor activity in patients will really help,” Hess says. “Since there aren’t many drugs that act at AMPA receptors, one of our goals is to identify drugs that change the ‘downstream’ effects of AMPA receptor activation. For example, we may be able to find other drug classes that change neuronal activity in the same way that AMPA receptor blockade changes activity.”

Posted on January 6, 2012 by Quinn Eastman in Neuro Leave a comment

mTOR inhibitors gaining favor for breast cancer treatment

This week, breast cancer researchers have been reporting encouraging clinical trial results with the drug everolimus at the San Antonio Breast Cancer Symposium. Everolimus is a mTOR inhibitor, first approved by the FDA for treatment of kidney cancer and then for post-transplant control of the immune system.

Ruth O’Regan, MD, director of the Translational Breast Cancer Research Program at Winship Cancer Institute, has led clinical studies of everolimus in breast cancer and has championed the strategy of combining mTOR inhibitors with current treatments for breast cancer.

She recently explained the rationale to the NCI Cancer Bulletin:

She views the combination therapy as a potential alternative to chemotherapy for treating ER-positive advanced breast cancer when hormonal therapies have stopped working.

When resistance to hormonal therapies occurs, Dr. O’Regan explained, additional signaling pathways become activated. Unlike chemotherapy, which targets rapidly dividing cells, mTOR inhibitors are an example of the kind of treatment that may block growth-promoting signaling pathways.

Currently, Winship researchers are examining a combination involving everolimus and the EGFR inhibitor lapatinib for “triple-negative” breast cancer, a particularly aggressive and difficult-to-treat variety.

Posted on December 9, 2011 by Quinn Eastman in Cancer Leave a comment

DNA copying problems explain muscular dystrophy mutations

Geneticist Madhuri Hegde and her colleagues have a paper in the journal Genome Researchthat addresses the question: where do copy number variations come from?

Madhuri Hegde, PhD

Copy number variations (CNVs), which are deletions or duplications of small parts of the genome, have been the subject of genetic research for a long time. But only in the last few years has it become clear that copy number variations are where the action is for complex diseases such as autism and schizophrenia. Geneticists studying these diseases are shifting their focus from short, common mutations (often, single nucleotide polymorphisms or SNPs) to looking at rarer variants such as CNVs. A 2009 discussion of this trend with Steve Warren and Brad Pearce can be found here.

Hegde is the Scientific Director of the Department of Human Genetics’ clinical laboratory. Postdoctoral fellow Arun Ankala is the first author. In the new paper, Ankala and Hegde examine rearrangements in patients’ genomes that arose in 62 clinical cases of Duchenne’s muscular dystrophy and several other diseases. Mutations in the DMD gene are responsible for Duchenne’s muscular dystrophy.

The pattern of the rearrangement hints what events took place in the cell beforehand, and hint that a problem took place during replication of the DNA. The signature is a tandem duplication of a short segment next to a large deletion, indicating how the DNA was repaired.

The authors note that the DMD locus is especially prone to these types of problems because it is much larger than other gene loci. The gene is actually the longest human gene known on the DNA level, covering 2.4 megabases (0.08 percent of the genome.)

Replication origins are where the DNA copying machinery in the cell starts unwinding and copying the DNA. Bacterial circular chromosomes have just one replication origin. In contrast, humans have thousands of replication origins spread across our chromosomes. In the discussion, the authors suggest that DNA copying problems may also explain duplications and historically embedded rearrangements of the genome.

Posted on December 1, 2011 by Quinn Eastman in Neuro Leave a comment

Scientists identify trigger for glowing plankton

Have you ever waded or paddled through ocean water in dim light, and found that your actions caused the water to light up?

Susan Smith, PhD

Single-celled plankton called dinoflagellates are responsible for this phenomenon. Almost 40 years ago, scientists studying bioluminescence (light emitted by living things) proposed a mechanism by which physical deformation of the cell could lead to a trigger of the flash.

Susan M.E. Smith, a research assistant professor in David Lambeth’s laboratory in Emory’s Department of Pathology and Laboratory Medicine, recently was first author on a paper in PNAS identifying a molecule that scientists have long believed to be the key to this mechanism. The paper is the result of a collaboration with Tom DeCoursey’s laboratory at Rush University in Chicago.

The mechanism for the trigger, first envisioned by co-author Woody Hastings, works like this. It is known that acidic conditions activate luciferase, the enzyme that generates the light. Part of the dinoflagellate cell, the vacuole, is about as acidic as orange juice. Normally the acidity within the vacuole is kept separate from the luciferase, which is found in pockets on the outside of the vacuole called scintillons.

Proton channels are needed to trigger bioluminescence. Illustration courtesy of the National Science Foundation, which supported Smith's research

Now something is needed to let acidity (that is, protons) pass from the vacuole to the scintillons. That something is a proton channel: a protein that acts as a gate in the membrane, opening in response to electrical changes in the cell. Smith and her collaborators identified a proton channel called kHV1 that has unique properties: it lets protons flow in the right direction for the trigger to work! They studied kHV1 by inserting the dinoflagellate gene that encodes it into mammalian cells and probing its electrochemical properties, which are distinct from other proton channels.

The authors write: “Whereas other proton channels apparently evolved to extrude acid from cells, kHV1 seems to be optimized to enable proton influx.”

The gene they found actually comes from a type of dinoflagellate that does not flash: K. veneficum, which feeds on algae and sometimes forms harmful blooms that kill fish. They propose that it uses acid influx to aid in capturing or digesting its prey.

“Hastings’ prediction led us to look for this kind of channel, we found it in a related organism, and it had the right properties to fit the prediction,” Smith says, and adds that her team has since found a similar gene in flashing dinoflagellates. She says studying the proton channel may give clues to ways to control harmful dinoflagellates, as well as help scientists understand how plankton respond to greater ocean acidity.

Proton channels are found in humans too. In fact, the same kind of molecule that triggers plankton flashing in the ocean helps human white blood cells produce a bacteria-killing burst of bleach. They are also involved in allergic reactions and in sperm maturation.

Smith is co-author on a paper that is in the journal Nature this week, exploring the selectivity of the human version of kHV1. Smith says that her interest in proton channels grew out of her work on Nox enzymes (which produce the bacteria-killing bleach) with Lambeth.

“I got interested in the proton channel because its function is necessary for peak Nox performance in human phagocytes. We started a little side project on the human proton channel that kind of blossomed,” she says. Her collaboration with DeCoursey uses “evolutionary information to get at the function of these channels in general.”

Posted on October 25, 2011 by Quinn Eastman in Uncategorized 2 Comments

COX-2 and epilepsy: it’s complicated

How much is the development of epilepsy like arthritis?

More than you might expect. Inflammation, or the overactivation of the immune system, appears to be involved in both. In addition, for both diseases, inhibiting the enzyme COX-2 initially looked like a promising approach.

Ray Dingledine, PhD

COX-2 (cyclooxygenase 2) is a target of traditional non-steroid anti-inflammatory drugs like aspirin and ibuprofen, as well as more selective drugs such as Celebrex. With arthritis, selectively inhibiting COX-2 relieves pain and inflammation, but turns out to have the side effect of increasing the risk of heart attack and stroke.

In the development of epilepsy, inhibiting COX-2 turns out to be complicated as well. Ray Dingledine, chair of pharmacology at Emory, and colleagues have a new paper showing that COX-2 has both protective and harmful effects in mice after status epilepticus, depending on the timing and what cells the enzyme comes from. Status epilepticus is a period of continuous seizures leading to neurodegeneration, used as a model for the development of epilepsy.

Postdoc Geidy Serrano, now at the Banner Sun Health Research Institute in Arizona, is first author of the paper in Journal of Neuroscience. She and Dingledine were able to dissect COX-2’s effects because they engineered mice to have a deletion of the COX-2 gene, but only in some parts of the brain.
They show that deleting COX-2 in the brain reduces the level of inflammatory molecules produced by neurons, but this is the reverse effect of deleting it all over the body or inhibiting the enzyme with drugs.

Four days after status epilepticus, fewer neurons are damaged (bright green) in the neuronal COX-2 knockout mice.

Dingledine identified two take-home messages from the paper:
First, COX-2 itself is probably not a good target for antiepileptic therapy, and it may be better to go downstream, to prostaglandin receptors like EP2.
Second, the timing of intervention will be important, because the same enzyme has opposing actions a few hours after status epilepticus compared to a couple days later.

More of Dingledine’s thinking about inflammation in the development of epilepsy can be found in a recent review.

Posted on October 19, 2011 by Quinn Eastman in Neuro Leave a comment

Drug discovery: shifting from brain growth factors to insulin

Earlier this year, the FDA put limitations on some anti-diabetic drugs because of their cardiovascular risks. The prevalence of diabetes in the United States continues to increase and is now above 8 percent of the population, so the need for effective therapies remains strong.

Keqiang Ye, PhD

Pathologist Keqiang Ye and colleagues have a paper in the Journal of Biological Chemistry describing their identification of a compound that mimics the action of insulin. This could be the starting point for developing new anti-diabetes drugs.

The new research is an extension of the Ye laboratory’s work on TrkA and TrkB, which are important for the response of neurons to growth factors. Ye and Sung-Wuk Jang, a remarkably productive postdoc who is now an assistant professor at Korea University, developed an assay that allowed them to screen drug libraries for compounds that directly activate TrkA and TrkB. This led them to find a family of growth-factor-mimicking compounds that could treat conditions such as Parkinson’s disease, depression and stroke.

Since TrkA/B and the insulin receptor are basically the same kind of molecule — receptor tyrosine kinases– and use some of the same cellular circuitry, Ye and Jang’s assay could also be used with the insulin receptor. Kunyan He and Chi-Bun Chan are the first two authors on the new paper. They report that the compound DDN can make cells more sensitive to insulin and improve their ability to take up glucose. They show that DDN (5,8-diacetyloxy-2,3-dichloro-1,4- naphthoquinone) can lower blood sugar, both in standard laboratory mice and in obese mice that serve as a model for type II diabetes.

Ye reports that he and his colleagues are working with medicinal chemists to identify related compounds that may have improved efficacy and potency.

“I hope in the near future we may have something that could replace insulin for treating diabetes orally,” he says.

Posted on October 7, 2011 by Quinn Eastman in Uncategorized Leave a comment

Redirecting beta-amyloid production in Alzheimer’s

Pharmacologist Thomas Kukar is exploring a strategy to subtly redirect the enzyme that produces beta-amyloid, which makes up the plaques appearing in the brains of Alzheimer’s patients.

Thomas Kukar, PhD

Preventing beta-amyloid production could be an ideal way to head off Alzheimer’s, but the reason why a subtle approach is necessary was illustrated last year by disappointing results from a phase III clinical trial. The experimental drug semagacestat was designed to block the enzyme gamma-secretase, which “chomps” on the amyloid precursor protein (APP), usually producing an innocuous fragment but sometimes producing toxic beta-amyloid.

Gamma-secretase also is involved in processing a bunch of other vital proteins, such as Notch, central to an important developmental signaling pathway. Scientists suspect that this is one of the reasons why trial participants who received semagacestat did worse on cognitive/daily function measures than controls and saw an increase in skin cancer, leading watchdogs to halt the study.

While a postdoc at Mayo Clinic Jacksonville and working with Todd Golde and Edward Koo, Kukar identified compounds – gamma-secretase modulators or GSM’s — that may offer an alternative.

“We are looking at a strategy that’s different from global gamma-secretase inhibition,” he says. “The approach is: don’t inhibit the enzyme overall, but instead modify its activity so that it makes less toxic products.”

Gamma-secretase chomps on amyloid precursor protein, and how it does so determines whether toxic beta-amyloid is produced. It also processes several other proteins important for brain function.

This line of inquiry started when it was discovered that some anti-inflammatory drugs also could reduce beta-amyloid production. Then, the crosslinkable probes Kukar was using to identify which part of the gamma-secretase fish was doing the chomping ended up binding the bait (APP). This suggested that drugs might be able to change how the enzyme acts on one protein, APP, but not others.

Now an assistant professor at Emory, he is examining in greater detail how gamma-secretase modulators work. Two recent papers he co-authored in Journal of Biological Chemistry show 1) how the proteins that gamma-secretase chews up are “anchored” in the membrane and 2) how selective GSM’s can be on amyloid precursor protein.

Although clinical studies of a “first generation” GSM, tarenflurbil, were also stopped after negative results, Kukar says GSM’s still haven’t really been tested adequately, since researchers do not know if the drugs are really having an effect on beta-amyloid levels in the brain. Newer compounds coming through the pharmaceutical pipeline are more potent and more able to get into the brain. While looking for more potent GSM’s is critical, Kukar says it’s equally as important to understand how gamma-secretase works to understand its biology.

Posted on September 16, 2011 by Quinn Eastman in Neuro Leave a comment

Blue pill or red pill? Brains need both for memory consolidation

In the 1999 film The Matrix, the character Neo is offered a choice between a blue pill (to forget) and a red pill (to remember). If only neuroscience was that simple! It may be that neurons need both red and blue, possibly an elaborate dance of molecules, for a fragile memory to lodge itself in the brain.

Neuroscientists Kimberly Maguschak and Kerry Ressler provide a glimpse into this process with their recent paper in the Journal of Neuroscience.

Ressler is both a psychiatrist and a Howard Hughes Medical Institute-supported researcher with a laboratory at Yerkes National Primate Research Center. Maguschak completed her doctorate at Emory and is now a postdoc with Guoping Feng at MIT.

The research is a follow-up on their work probing the role of beta-catenin in fear memory formation. We previously described this protein as acting “like a Velcro strap”, attaching cells’ internal skeletons to proteins on their external membranes that help them adhere to other cells. If brain cells need to change shape and form new connections for memories to be consolidated, we can see how this kind of molecule would be important.

Beta-catenin is also central to a signaling circuit that maintains stem cells and prods an embryo to separate into front and back or top and bottom. This circuit is called “Wnt” (the name is a fusion of the fruit fly gene wingless and a cancer-promoting gene discovered in mice, originally called Int-1).

Maguschak and Ressler wanted to assess the role Wnt signals play in learning and memory. The model system was the same as in their previous work: if mice are electrically shocked just after they hear a certain tone, they gradually learn to fear that tone, and they show that fear by freezing.

Kerry Ressler, MD, PhD

Maguschak saw that in the amygdala, a part of the brain important for fear responses, Wnt genes are turned down during the learning process temporarily but then come back on. If the mice only hear the tone or only get the shock, the genes’ activities don’t change significantly.

She then introduced proteins that perturb Wnt signaling directly into the amygdala. Extra Wnt injected before training, while it didn’t stop the mice from learning to fear the tone, made that training less likely to “stick.” Two days later, the mice that received Wnt didn’t seem to fear the tone as much.

Here’s the possibly confusing part: a Wnt inhibitor also impaired fear memory consolidation. In effect, both blue and red pills actually interfered with how well memories endured. The authors suggest this is because Wnt signals have to be turned down during fear memory formation but then turned back up so those memories can solidify. The Wnt signals seem to go along with the adhesive interactions of beta-catenin. It looks like beta-catenin’s stickiness also needs to be tuned down and then back up.

The off-then-on-again requirement Maguschak and Ressler observe is reminiscent of results from cell biologist James Zheng’s lab. He and his colleagues saw that the actin cytoskeleton needed to be weakened and then stabilized during long-term potentiation, an enhancement of connections between neurons thought to lie behind learning and memory.

Several laboratories have identified potential drugs that modify beta-catenin/Wnt. These new results suggest that the timing of when and how to use such drugs to enhance memory may critically important to consider, Ressler says.

“To interfere with memory formation after trauma or enhance memory formation in people with dementia, researchers will clearly need to attend to the full complexity of the dynamics of synaptic plasticity and memory,” he says.

A nifty link to an animation of Wnt signaling

 

Posted on September 14, 2011 by Quinn Eastman in Neuro Leave a comment

Personalized Medicine Day in Georgia

Governor Nathan Deal was joined by Ambassador Andrew Young, Georgia State Representative Calvin Smyre and Leroy Hood, founder of the Institute of Systems Biology, in formally proclaiming September 1, 2011 Personalized Medicine Awareness Day in the State of Georgia.

Georgia Governor Nathan Deal presents Morehouse School of Medicine’s Dean and Executive Vice President, Valerie Montgomery Rice, MD, with a state proclamation declaring Sept. 1, 2011 Personalized Medicine Awareness Day in Georgia.

The event at Morehouse School of Medicine (MSM) was sponsored by Georgia Bio; the Atlanta Clinical & Translational Science Institute (ACTSI, which is funded by the NIH and led by Emory University with partners MSM and Georgia Tech); and Iverson Genetics, Inc.

“The collaboration within the ACTSI between these three research universities is an important undertaking and an example of how it should be done,” remarked Governor Deal as he kicked off the day’s program.

A visionary in the personalized medicine field, Dr. Hood developed the DNA gene sequencer and synthesizer and the protein synthesizer and sequencer – four instruments that paved the way for the successful mapping of the human genome.

During his keynote address he proposed a revolution in medicine.  P4 Medicine – Predictive, Preventive, Personalized and Participatory – is a proactive (instead of a reactive) approach to medicine. The paradigm change will drive radical changes in science.

For P4 medicine to succeed, a cross-disciplinary culture with team science and new approaches to educating scientists, as is done through the ACTSI, has to take place. Dr. Hood predicts the human genome will be part of individual medical records in 10 years.

Leroy Hood, MD, PhD

“The vision of P4 medicine is that each patient will be surrounded by a virtual cloud of billions of data points. Advances in science and technology will reduce this enormous data dimensionality to simple hypotheses about human health and disease,” says Hood.

“The ultimate outcome is to create individualized patient disease models that are predictive and actionable. The shift to P4 Medicine will also require societal changes.”

Personalized Medicine Awareness Day celebrated the first-of-its-kind personalized medicine study, approved by the Centers for Medicare and Medicaid Services. The study will determine the utility of genetic testing in calculating doses and reducing the incidence of adverse events associated with the initiation of Warfarin therapy. Warfarin is the world’s leading anti-blood clotting drug.

Researchers hope the study will provide data to demonstrate that individualizing treatment can improve patient safety and reduce healthcare costs, says Dean Sproles, CEO of Iverson Genetics, Inc., which is collaborating in the study with MSM and the ACTSI.

Governor Deal congratulated the ACTSI for leading the landmark Warfarin study with Iverson and is “proud that Georgia will be leading the effort.”

The Warfarin Study is led by ACTSI Senior Co-Principal Investigator Elizabeth Ofili, MD, MPH, director of the Clinical Research Center, chief of cardiology and associate dean for clinical research at MSM, and will engage 50 sites across the country and 7,000 participants. The first participant was recently enrolled at Grady Memorial Hospital.

“This study should help us understand how to use each patient’s genetic information to deliver a safer and more effective dose,” says Ofili.

Sproles noted, “The study is evidence of the growing role of genetics in helping doctors to develop optimal individual treatments for their patients.”

A panel including Emory medical leaders David Stephens, Fred Sanfilippo and Kenneth Brigham discussed and addressed questions like how to communicate ‘big science’ to the individual, how to move genetic testing to medical outcomes and who owns genome data.

“Personalized Medicine is the future,” stated Governor Deal. The presence of Governor Deal, Ambassador Young and Representative Smyre is a sign that policymakers are beginning to recognize that personalized medicine is not just a vision for better healthcare; it has the power to improve health and reduce healthcare costs.

Posted on September 9, 2011 by admin in Uncategorized 1 Comment
« Previous   1 2 3 4 5 6 7 8 9 10 ... 23 24   Next »