MikeLigalig.com

Singing Lessons: How to Break Away From Bad Habits

Believe it or not, the hardest thing to do is to break away from bad habits. Only those who succeed in doing this will become better singers. I've had calls from musicians (voice trained by me) complaining their vocal performance on stage had gone downhill and that they want to come back for more lessons. When I dig deep, it quickly becomes apparent that they've gone back to their old ways; I see my OWN student competing in a reality TV singing competition and doing NOTHING I taught them to do. It's like they never went for vocal training at all. The whole shebang: bad pitching, yelling and screaming their way around a song-I mean the whole flaming caboodle LIVE on TV. Unbelievable! I've seen my own students fall apart in the recording studio. Everything they've learned goes suddenly out the window. They even forget to BREATHE before a note. How bad habits die hard, hey? This article will investigate the perils of regressing into your old singing habits and how to break the cycle.

Source: Singing Lessons: How to Break Away From Bad Habits

De-Clutter to Energize Yourself and Your Life

If you feeling stuck in some area of your life and want to release old baggage that's weighing you down, it is likely a good time to de-clutter. The act of de-cluttering is a great way to let go and release stale energy so that you can welcome new opportunities and experiences into your life. Letting go is also a fundamental step in honoring your personal growth. There are 3 areas that you can de-clutter to energize yourself and your life.

Source: De-Clutter to Energize Yourself and Your Life

Silence Your Inner Critic and Go for Your Dreams

It is time for you to silence your inner critic and go for your dreams. Your dreams are meant to be achieved, and you were born to succeed. It is your moment to dream big and achieve big. You deserve it.

Source: Silence Your Inner Critic and Go for Your Dreams

Fulfill Your Purpose As a Life Coach

Fulfilling your purpose as a life coach can change your life and the lives of others. You have what it takes to be an amazing life coach, and you deserve to do what you love to do. If you are being led to life coaching now is your time to pursue your purpose.

Source: Fulfill Your Purpose As a Life Coach

Coaching - A Significant Program For Managers

This is an article identifying some of the reasons why it is important for an organization to consider coaching for career in their training programs. It points out how coaching can be a helpful tool in increasing performance in the overall.

Source: Coaching - A Significant Program For Managers

Serve the One You're With

Recently, several of the yoga coaches I've trained were talking about how their class and client numbers are down. I shared a few tips with them about how to bring those numbers back up coming from a dharmic place of service. If that's also been a challenge for you, this might help.

Source: Serve the One You're With

Coaching Corporate Executives to Maintain the Start-Up Entrepreneur Mentality

In late October of 2012 Microsoft came out with its new tablet computer called The Surface RT. It is a tablet which has a touchscreen, but also a Windows operating system which allows you to use Microsoft office and Word programs. In fact, you can use the full version of the Microsoft Word program. When Bill Gates was interviewed, he suggested that not only was the Surface RT going to be a huge success, but also that Steve Ballmer was going to start operating the company with more of a startup mentality.

Source: Coaching Corporate Executives to Maintain the Start-Up Entrepreneur Mentality

Coaching Controversial Radio Hosts

If you want to be great in radio, you need to be controversial and hard-hitting, but you also need to know when to stop. You have to be careful with what you say, and maintain a little bit of political correctness. That doesn't mean you can't rile up your audience a little bit, and let the callers calling in do all the talking with all the harsh words. If you do that you still get your point across without making statements that might be used out of context later on to destroy your career, reputation, or get you fired from the network because they can't risk losing sponsors or advertisers. Welcome to the world of real-world politics I say.

Source: Coaching Controversial Radio Hosts

Finding the Right Women's Coach For Your Needs

There are a lot of women's coaches to choose from. All of them have their own experiences, backgrounds, and training that shaped their own lives and career in order to help others. It is key to consider these factors as you choose a women's coach for your needs.

Source: Finding the Right Women's Coach For Your Needs

What It Takes to Achieve GREATNESS

It's not enough to be the best... when you have the ability to be GREAT. From a societal perspective, there is this stereo-typical notion of what amounts to greatness in an individual. If you were to ask any reasonably informed person to name a number of great individuals they might mention people like; Winston Churchill, Nelson Mandela, Mother Teresa, Tommy Douglas, Terry Fox, Bill Gates. You will not get any argument from anyone with those choices?

Source: What It Takes to Achieve GREATNESS

Creating Legendary Leadership With The Enneagram

Do you want to trailblaze your leadership effectiveness and results? This month I'm writing about Legendary Leadership, how understanding the leadership style of each Enneagram type creates transformative, conscious, empowered and inspired leadership at work.

Source: Creating Legendary Leadership With The Enneagram

Turning Weaknesses Into Strengths

Dealing with the question: "What is your main weakness?" at interview can be tricky, but with the proper mindset and preparation you can use it to show your strengths...

Source: Turning Weaknesses Into Strengths

Practicing Compassion And Mindfulness

How to plant seeds of mindfulness and compassion that you hope will sprout in women's minds. This is how the journey to wellness begins to take over their toxicity.

Source: Practicing Compassion And Mindfulness

It's Not Whom You Coach, It's The Way That You Coach Them

Perhaps, you noted that catchy branding jingle that HP was running for its printers and peripherals in the fall of 2012. It goes something like this; "it's not what you, it's the way that you do it," and it does stick in your head, plus they were playing it all day long for weeks on end on CNBC financial news during the day. Well, the other day I was perusing some online coaching articles and there was someone selling their coaching services to help coaches learn how to coach and make money doing it.

Source: It's Not Whom You Coach, It's The Way That You Coach Them

How to Coach Clients in Humanitarian Organizations

The other day, someone had contacted me and wanted me to help them get set up in their own nonprofit group which would be engaged in humanitarian efforts. I tried to talk them out of it because I've learned a lot and have had a lot experience with that over the years. It's not as easy as it looks, and yes, while everyone does want to save the world, sometimes the world doesn't want to save itself. Worse, there are often gigantic arguments between various political groups, and humanitarian organizations on how to go about what is necessary to help the people in those regions of the world.

Source: How to Coach Clients in Humanitarian Organizations

How You Can Benefit From a Woman's Coach

A woman's coach exists to motivate and help women improve their business and personal lives. As women advance in business, they have found coaching to be an integral part of their success.

Source: How You Can Benefit From a Woman's Coach

3 Reasons Women Coaches Struggle to Achieve Their Money Goals

If you are a woman coach struggling to achieve your money goals what needs to change? If you are struggling you don't have to be. You have the inner power to shift the direction of your money flow.

Source: 3 Reasons Women Coaches Struggle to Achieve Their Money Goals

Are You Busy, Really Busy, or Are You Abundant?

I had an epiphany the other day during my run. I was thinking about all the things that I 'had' to get done that day. While going through my list in my head I heard a familiar tape playing in the back of my mind; "I'm so busy!

Source: Are You Busy, Really Busy, or Are You Abundant?

How to Refocus and Change Your Attitude

Attitude is a small thing but it can change your life if you get it going in a more positive light. See, a lot of people walk around mad at the world and angry and bitter and going in the same direction everyday.

Source: How to Refocus and Change Your Attitude

How to Deliver the Feedback Element of Your Coaching Program

Email: If you are delivering the feedback element of your coaching program via email, create an email account you will use exclusively for your coaching clients. Give that email address to your clients, let them know when you are available for email questions (for example, Monday through Thursday from 9-4), and simply respond to their emails as they come in.

Source: How to Deliver the Feedback Element of Your Coaching Program

How to Create Coaching Program Lessons

There are 2 primary ways to create coaching program lessons. One way is to create them ahead of time, much as you would create an information product to sell. For example, if you are going to launch a 13 week coaching program you would create 13 coaching lessons ahead of time. Another way is to create the lessons as your clients will require them. For example, if you have a 13 week coaching program, you create the first lesson the first week, the second lesson the second week, and so on. If you create lessons as you need them in this way, the first time you offer coaching, you can use the now-created lessons for future classes in your coaching program.

Source: How to Create Coaching Program Lessons

11 Forgotten Laws - Review

Have you ever wondered why the Law of Attraction doesn't work for you or only works sometimes but not all of the time? The answer is because the Law of Attraction has many other laws that were never mentioned in the movie The Secret. In fact, there are 11 Forgotten Laws that the movie never even talks about. These missing or forgotten laws hold the key to your success.

Source: 11 Forgotten Laws - Review

How and Why to Avoid Temptation

Due to the nature of their work, therapists and attorneys often find themselves alone with their patients/clients. In theory, that shouldn't present ethical issues, but in reality, human nature can come into play and on occasion, it can be tempting to initiate more personal relationships.

Source: How and Why to Avoid Temptation

4 Steps on How to Apply The 11 Forgotten Laws

The 11 Forgotten Laws is a highly acclaimed self improvement program. Find out 4 tips you can learn from The 11 Forgotten Laws to help you apply the Law of Attraction and manifest more success.

Source: 4 Steps on How to Apply The 11 Forgotten Laws

Laser Hair Removal Easy And Safe

There are many methods to remove unwanted hair. None are completely permanent, but laser hair removal is the most effective treatment available for the removal of unwanted facial and body hair.

Source: Laser Hair Removal Easy And Safe

Law of Attraction - Why It Does Not Work For You

The law of attraction seems easy for some while others are having a hard time with it. Others wonder why it works for others and not to them. The reason for is simple. You become impatient and you do not truly believe that you will get what you want.

Source: Law of Attraction - Why It Does Not Work For You

Ways of Attracting Luck in Life

People have different perceptions about luck but one thing is for sure, luck is real. That is the only way to explain why some people are consistently lucky unlike others. Many times you may have been thinking of how to attract luck, how to influence the outcome of our day to day activities so that they may favor you.

Source: Ways of Attracting Luck in Life

Watch Me - I'll Figure It Out!

This is an inspiring experience from my client, Aimee, which reads like a how-to manual for getting what you want.? As you read Aimee's experience, notice how she focuses on her desire with clarity and determination.

Source: Watch Me - I'll Figure It Out!

The Secret - What Is It?

A simple introduction to what 'The Secret' is and how you can start to make it work for you. Thought power is everything, don't underestimate it!

Source: The Secret - What Is It?

Law Of Attraction: Resist Insecurities For A Real Life

A little insecurity is considered normal but to experience insecurities to the extent that they cripple you from coping with problems or situations can be potentially dangerous. Insecurities forces you to operate from a place that is imaginative only in your mind. Rather than feeling powerless, you should know that you can turn the law of attraction to your advantage. It is never too late to change your attitude in life.

Source: Law Of Attraction: Resist Insecurities For A Real Life

How Brainwave Entrainment Can Help in Manifestation

Looking to manifestation as a way of attracting the life you desire? Brainwave entrainment can be used to assist you in this process.

Source: How Brainwave Entrainment Can Help in Manifestation

7 Falsehoods of "The Secret"

If the popular movie and book "The Secret" would work as advertised, most people watching it would be millionaires by now. That hasn't happened. Learn why the principles in "the Secret" lead to failure and how to remedy them.

Source: 7 Falsehoods of "The Secret"

Luck And The Single Chance

I would like to begin this article with a realistic thought: There is not any such thing as luck, and since there is such thing as temporary failure and ultimate success, there is always more than a single chance to succeed in existence. Let me explain.

Source: Luck And The Single Chance

How Gratitude Ignites the Law of Attraction

The law of attraction states "like attracts like". What better or faster way is there to attract all that you desire than the state of gratitude? (hint: none:) Read on to found out how to instantly feel more grateful and be a magnet to all your heart's desires.

Source: How Gratitude Ignites the Law of Attraction

Why Karma Works

The law of cause and effect or attraction works like this: It is the most basic law in existence without matter to any other law anyone else would make in any way because it underpins everything from the most basic physics to the most complex political law. That is reality whether we like it or not.

Source: Why Karma Works

Sinking Thoughts Then Rising Thoughts, The Power Of The Mind

Life rises and sinks according to the deepest nature of our thoughts and actions, in that order. With that said, I will fully explain myself.

Source: Sinking Thoughts Then Rising Thoughts, The Power Of The Mind

The Law of Attraction Put to the Test

The author describes the construction of a rigorous test to prove to his own satisfaction whether there could be any validity to the law of attraction. This test had to be sufficiently specific to avoid the trap of poor experiment design while also being plausible and difficult to obtain. The result was a multi-year test worth reading to see if you agree with the author's end conclusions.

Source: The Law of Attraction Put to the Test

My Healing With the Law of Attraction

In 2007 I was diagnosed with Meniere's Disease - an inner ear disorder that causes violent attacks of tinnitus, which is so loud that I was virtually deaf in the affected ear. The tinnitus then causes dizziness, which results in vomiting. Attacks are unpredictable and can come at any time anywhere.

Source: My Healing With the Law of Attraction

The Law of Attraction: The Secret to Raising Your Vibration and Manifesting Your Dreams

Discovering my spirituality many years ago - I remember my best friend at the time explaining to me, be around the places you want to live, draw a description of the house you want to live and find pictures of the person who looks like the one you want to be with. It's about being around, feeling and giving clear intentions of what you want to attract into your life.

Source: The Law of Attraction: The Secret to Raising Your Vibration and Manifesting Your Dreams

Try To Be Friendly Towards Everyone

There's only one caveat about walking around and smiling at everyone. Like so many other things in life, everything's fine in moderation. When you pass someone in a corridor, just a quick nod and a smile are quite sufficient.

Source: Try To Be Friendly Towards Everyone

What Woman Really See When They Look At Men

Whether we will admit it or not, the majority of men dress to attract the opposite sex. It is a fact that much of the stuff we do in public is more bait than anything else. Since this is the case, it would behoove men to be aware of how alluring their bait is to their prey. So what is it that women are actually seeing when they look at men? Are men winning women over with their fashion sense or is something much deeper going on?

Source: What Woman Really See When They Look At Men

Curbing Self Sabotage

Often we are our own worst enemies when it comes to achieving our desires. When we get close to achieving them we often sabotage ourselves by falling into the trap of believing our doubts about ourselves and our abilities.

Source: Curbing Self Sabotage

The Close Sibling Of A Good Thought

I will not mince words. The close brother or sister of a good thought is a productive action. Now, to spend the rest of this article explaining my point of view.

Source: The Close Sibling Of A Good Thought

The Benefits of Letting Go of Negative Emotion

Once you have committed to a path to your greatest desire and you are doing something every day to practice, or develop a new skill, it is important to just let go of all of the limiting thoughts that are only serving to get in the way of your Dream, or distract you from moving forward. Here are tips to letting go, why it's important and how to make a new positive state of mind work for your greatest benefit.

Source: The Benefits of Letting Go of Negative Emotion

Empathy and Emotional Intelligence

Being empathetic is simply putting yourself in the other person's shoes. It is "the ability to feel what someone else is feeling." Empathy and Emotional Intelligence work together to produce long-lasting relationships.

Source: Empathy and Emotional Intelligence

Become Debt Free With The Law Of Attraction

In these difficult economic times many people are experiencing financial difficulties. The threat to jobs caused by companies downsizing and laying people off is also cause for concern with a lot of people. Some people have such large debts and the financial burden is so great that they do not see a way out. However, by studying the Law of Attraction you will learn and understand that you are the master of your own destiny. You will learn that you have the power to become debt free and bring about the lifestyle of your dreams. You do not have to wait for external forces to dictate what happens in your life.

Source: Become Debt Free With The Law Of Attraction

What Makes One Attractive To Others

Have you ever let go of your body at one point in life? if you let go, you no longer watch what you eat, you can no longer fit into your clothes like you once did, you start to feel like you are looking less and less attractive to the opposite sex. Well the remedy is to go back to the basics.

Source: What Makes One Attractive To Others

The Secret to Belief

On a scale of one to ten, what is your level of belief in yourself? Many people think that they are positive because they consciously choose positive words and say, "I believe". But true belief is a feeling.

Source: The Secret to Belief

Create Your Life On Purpose: Balancing Action With LOA II - Winning the Waiting Game

An illuminative series that aims to equip readers with the insights they need, and tips for the practical steps of faith they can take, to move forward into their dream life. This series will illustrate from my own personal life experience how I developed a confidence in and a working knowledge of spiritual wisdom, understanding and science (also known as the Law Of Attraction) and am progressively creating the life of my dreams.

Source: Create Your Life On Purpose: Balancing Action With LOA II - Winning the Waiting Game

Congratulations - It's a Mouse!

The magic behind wondering what positive outcome will result from a negative experience is the understanding that you have the power to attract either positive or negative circumstances from any situation. In fact, focusing on the blessing in disguise actually brings the blessing.

Source: Congratulations - It's a Mouse!

The Deeper Secret to the Law of Attraction

The Law of Attraction means that we are creating our own reality with our consistent thoughts. In other words, what we experience in life depends on what we project on many levels. Of course, this is just the tip of the iceberg.

Source: The Deeper Secret to the Law of Attraction

Attract Abundance By Getting Rid Of "Need"

Getting rid of the "needy" feelings opens the doors to so much. It allows your energy to flow freely through you, which will attract much abundance to you. This is very common with thoughts towards relationships and money.

Source: Attract Abundance By Getting Rid Of "Need"

How To Dress And Get Noticed

The clothes make the man statement has been used for decades. But it is never more true than in this century. People from all walks of life have been given audience in places and with people who they otherwise would have never gotten the opportunity had it not been for their outward appearance. This platform has been used to rise to the level of dignitaries as well as those doing an expose where they needed to get into less than desirable environments by using clothes to make themselves look down trodden or streetwise.

Source: How To Dress And Get Noticed

Create Your Life On Purpose: Balancing Action With the Law of Attraction

This is an introductory article for an illuminative series that aims to equip readers with the insights they need, and tips for the practical steps of faith they can take, to move forward into their dream life. This series will illustrate from the author's personal life experience how she developed a confidence in and a working knowledge of spiritual wisdom, understanding and science (also known as the Law Of Attraction) and is progressively creating the life of her dreams.

Source: Create Your Life On Purpose: Balancing Action With the Law of Attraction

What Comes Around - Goes Around

All energies that are sent out will boomerang back to the sender. What has already come around will go around and vice-versa. Why does karma happen?

Source: What Comes Around - Goes Around

The Law Of Attraction - How Does It Affect Us?

The law of attraction is fun to learn and use because you are always watching and waiting expectantly for your dreams to become a reality. You can use this law to create the future you want. The law of attraction is working in your life right now, whether you know it or not. You attract the things that are happening around you. When you become aware of the law of attraction and how it works, you can use it to attract what you want.

Source: The Law Of Attraction - How Does It Affect Us?

The Law of Attraction and Your Home Business

The Law of Attraction. Your home business. They seem to be at completely opposite ends of one's life. But they're not. When implemented, The Law of Attraction will bring greater sales, happier customers, and a greater sense of purpose to you and your home business.

Source: The Law of Attraction and Your Home Business

Creating a More Popular and Likeable Personality

This article teaches you how to attract people. It also points out great ways to be a nice person.

Source: Creating a More Popular and Likeable Personality

Law Of Attraction: Accelerate It With Good Old Exercise

Everyone exercise as part of a healthy lifestyle. We all know that exercise is good for boosting our metabolism, losing weight and maintaining strong bones. However, does anyone know that exercise can be a great accelerator for the law of attraction?

Source: Law Of Attraction: Accelerate It With Good Old Exercise

Pain, Any Sort of Death And What I Am Convinced Is Beyond Both Conditions

To get beyond pain, death and any sort of loss, you have to stop it from being any sort of deterrent to your rational actions in your life, present or future. You can do nothing about the past, but learn from it to do better in the present and future.

Source: Pain, Any Sort of Death And What I Am Convinced Is Beyond Both Conditions

Is Indoor Tanning Safe? 4 Things You Must Know

Indoor tanning is the alternative to the sun where thousands of people flock to the tanning salons to get that natural golden tan. They use tanning beds and booths equipped with several fluorescent bulbs. The bulbs emit UVA rays which are equivalent to the sun. Tanners love indoor tanning because they don't sweat or burn and can get a tan in as little as five minutes. More people are using indoor tanning because of too much over exposure to the sun's rays. People often wonder if indoor tanning is safe.

Source: Is Indoor Tanning Safe? 4 Things You Must Know

Why Too Much Sun Causes Health Problems

The sun throws off ultraviolet rays which can damage your skin causing premature aging and skin cancer. These UV rays penetrate the skin causing our cells to die turning to tumors and other types of cancer. Too much sun causes many health problems from overexposure to the UV rays damaging your DNA, posing other threats such as diabetes, and the fatal Melanoma skin cancer.

Source: Why Too Much Sun Causes Health Problems

What Are The Benefits Of Indoor Tanning?

Indoor tanning is the best way to get a fast tan safely. Thousands of sun tanners have switched because of the great benefits, such as getting a natural tan in five to fifteen minutes, no sweating or getting dehydrated from thirst. There are risks associated with both types of tanning, but indoor tanning has lower risks compared to sun tanning. You can tan all year around regardless of the weather but you can only tan in the sun during the summer.

Source: What Are The Benefits Of Indoor Tanning?

Is Indoor Tanning A Bad Health Risk?

People often ask if indoor tanning is a bad health risk, and the answer is no if you listen to the instructor, protect your skin with lotion and wear tanning goggles to protect your eyes. The rules for indoor tanning are the same as sun tanning; do not stay in the bed or booth too long or you'll get overexposed to the UV rays. You're only in the booth for five minutes and the bed for twelve to fifteen minutes.

Source: Is Indoor Tanning A Bad Health Risk?

How To Get A Tan And Protect Your Skin Against Premature Aging

You can get an indoor or outdoor tan and protect your skin against premature aging by using lotions keeping your skin soft and youthful looking. It takes a few minutes to apply the lotion saving you years of wrinkled skin before your years. If you don't use a good SPF 15 tanning lotion when sun tanning, your skin will dry out, flake and sunburn. It causes premature aging along with staying in the sun too long.

Source: How To Get A Tan And Protect Your Skin Against Premature Aging

How Tanning Lotion Helps Protect Against Skin Cancer

Tanning lotion helps protect against skin cancer by blocking the ultraviolet rays that penetrate all layers of the skin. 95% of UVA rays are emitted from the sun giving you a tan by increasing the melatonin in your body. 5% of UVB rays emitted from the sun causes sunburn and changes your DNA. The UV rays damage your skin cells causing premature aging and skin cancer. There are different types of skin cancer but Melanoma is fatal and can't be treated.

Source: How Tanning Lotion Helps Protect Against Skin Cancer

Why Indoor Tanning Is Safer Than Sun Tanning

Indoor tanning is safer than sun tanning because the sun's UV rays are not consistent, sometimes hotter than other times. It takes hours in the sun to get a nice tan, where you can get the same tan from five to twenty minutes with indoor tanning. The sun has UVA and UVB rays where the UVA rays give you the tan, but the UVB rays damages your DNA. Indoor tanning only uses UVA rays. It's easier to get exposed to these rays from the sun because it's so much hotter and you sweat, sunburn and can have a heatstroke.

Source: Why Indoor Tanning Is Safer Than Sun Tanning

Are You Protecting Your Skin From Sun Tanning?

Many people are not aware of the dangers of sun tanning getting exposed to the UV rays. They don't think about protecting their skin with tanning lotion to block these harmful rays. They cause skin cancer, heat strokes and premature aging. There are different types of skin cancer but Melanoma is fatal and many people die every year from it. Overexposure to the sun can cause a heatstroke that drains the moisture out of your body and if not treated quickly, you can die from it.

Source: Are You Protecting Your Skin From Sun Tanning?

The Difference in Tanning Lotions and Which Ones to Use

People who don't use tanning lotion are at risk for skin cancer and premature aging. It is imperative that you apply a good UV block lotion all over your skin before tanning indoors or outdoors. It helps to moisturize your skin keeping it from drying out and will give you a glowing natural tan. Years ago people used butter when sun tanning. It burned the skin and did not protect them against the ravage damages from the UV rays.

Source: The Difference in Tanning Lotions and Which Ones to Use

How To Get A Natural Spray Tan

More people are leaving the sun for indoor spray tanning. You can get a beautiful tan in just five minutes, where the sun takes hours. Spray tanning has become the best way to get a natural tan fast and easy. Most indoor tanners are using this method more than any other way of tanning. When you go to the tanning salon, you step in a private booth, turn on the nozzle and you're done in five minutes.

Source: How To Get A Natural Spray Tan

How To Attract Women Fast?

Hey man, what's up? Are you upset because you are single for a while and you just can't seem to find a nice lady? Damn man, don be so sad, as there are so many ways out there through which you will be able to find your sweetheart, that you will be blown away.

Source: How To Attract Women Fast?

A Simple Genetic Test for Kidney Disease

This technology relates to methods of diagnosing a predisposition to diseases that cause chronic kidney disease (CKD) and end-stage kidney disease (ESKD). Variations in a gene, non-muscle myosin IIA (MYH9), are associated with 79% of the risk of focal segmental glomerulosclerosis (FSGS), the disease that causes ESKD, in African Americans with HIV, and 56% of African Americans as a whole. The variants are also associated with a 2-3 fold increased risk for ESKD associated with hypertension. The variations are also present among European Americans, although they are less common.

A simple genetic screening test has been developed that identifies single nucleotide polymorphisms (SNP) and haplotypes in the non-muscle myosin gene MYH9. These variants confer genetic risk for the following kidney diseases: FSGS, collapsing glomerulopathy, HIV-associated nephropathy, hypertensive kidney disease, sickle cell nephropathy, lupus nephropathy, and possibly other kidney diseases.

Source: A Simple Genetic Test for Kidney Disease

Gene Signature for Predicting Solid Tumors Patient Prognosis

A method of predicting the prognosis of a patient diagnosed with hepatocellular carcinoma (HCC) or breast cancer by detecting expression of one or more cancer-associated genes and a method of identifying an agent for use in treating HCC.

A progressive sequence of somatic mutations and epigenetic changes of oncogenes or tumor suppressor genes are believed to cause tumor development. However, high genomic instability in tumors causes the accumulation of genomic aberrations that do not contribute to tumor progression. Therefore, it is important to distinguish between ?driver? mutations that are functionally important and ?passenger? mutations that do not provide a selective advantage to the tumor cells.

The current invention describes a driver gene signature for predicting survival in patients with solid malignancies, including HCC and breast cancer.  The gene signature includes 10 cancer-associated genes. The NIH researchers also found that these cancer-associated genes can be divided into two different groups. While a decrease in DNA copy number or mRNA expression of one group of genes is associated with poor prognosis in HCC tumors and breast cancer, such a decrease in the other group is associated with good prognosis. The inventors have also demonstrated that at least four of these cancer-associated genes are functional tumor suppressor genes. Thus, these genes may be molecular targets of HCC and breast cancer.

Source: Gene Signature for Predicting Solid Tumors Patient Prognosis

Novel Biomarkers for Alcohol-Induced Liver Disease

Alcohol-induced liver disease (ALD) is reversible if identified in the early stages, but early diagnosis is difficult with existing tools. One problem associated with developing a new diagnostic tool is the genetic background associated heterogeneity in physiological responses to chronic alcohol consumption.  The inventors of the present technology have solved this problem and have discovered background-independent novel biomarkers for ALD.  In the current studies, the inventors generated two genetically distinct lines of PPARalpha-null mice and evaluated the levels of urine metabolites after alcohol exposure.  The inventors have identified indole-3-lactic acid and phenyllactic acid as putative biomarkers for ALD.  Indole-3-lactic acid and phenyllactic acid levels were significantly elevated in both lines of PPARalpha-null mice after two to three months of alcohol administration.  The inventors had identified indole-3-lactic acid and phenyllactic acid to be background independent markers for ALD.

Source: Novel Biomarkers for Alcohol-Induced Liver Disease

Mouse-Derived NY-ESO-1 T Cell Receptor for Use in Immunotherapy

Scientists at the National Institutes of Health have developed a T cell receptor (TCR) derived from mouse T cells (i.e. murine TCR) that can be expressed in human T cells to recognize the cancer testis antigen (CTA), NY-ESO-1, with high specificity. This anti-NY-ESO-1 TCR has murine variable regions that recognize the NY-ESO-1 epitope and murine constant regions. The inventors performed in vitro studies comparing this murine NY-ESO-1 TCR with a previously developed human NY-ESO-1 TCR counterpart, which yielded promising clinical outcomes in patients with a variety of cancers. The murine TCR functioned similarly to the human counterpart in their ability to recognize and react to NY-ESO-1 tumor targets.

NY-ESO-1 is a CTA, which is expressed only on tumor cells and germline cells of the testis and placenta. CTAs are ideal targets for developing cancer immunotherapeutics, such as anti-CTA TCRs, because these TCRs are expected to target cancer cells without harming normal tissues and thereby minimize the harsh side effects associated with other types of cancer treatment. NY-ESO-1 is expressed on a wide variety of cancers, including but not limited to breast, lung, prostate, thyroid, and ovarian cancers, melanoma, and synovial sarcomas.  Thus, this technology should be applicable in adoptive cell transfer therapies for many types of cancer.

Source: Mouse-Derived NY-ESO-1 T Cell Receptor for Use in Immunotherapy

Exposing T cells to Fas Ligand (FasL)-Fas Receptor (FasR) Antagonists Withholds Differentiation and Increases Expansion Making T cells More Suitable for Use in Cancer Immunotherapy

NIH scientists have developed methods to make a better immunotherapy by exposing T cells to Fas ligand (FasL) or Fas receptor (FasR) antagonists and agonists. Researchers have found that FasL-FasR antagonists suppress T cell differentiation leaving them in a na?ve state. These T cells are a more ideal cell type for adoptive cell transfer therapies because they have not exhausted their effector functions and demonstrate greater proliferation, enhanced persistence and survival, and better activity against their target antigen when infused in vivo to treat cancer. Also, the prevention of T cell differentiation/effector function in vivo has implications for autoimmune diseases and syndromes. FasL-FasR agonists enhance T cell differentiation towards more effector-like cells. Enhancing the differentiation of T cells is expected to be useful in treating cell proliferation disorders, such as leukemias, lymphomas, or Wiskott-Aldrich syndrome.

FasL (or cluster of differentiation 95L) is a transmembrane protein in the tumor necrosis factor (TNF) family. FasR (or apoptosis antigen 1, CD95, or TNF receptor superfamily member 6) is a transmembrane protein belonging to the TNF receptor/nerve growth factor receptor superfamily. Normally, when FasL binds to FasR, a cell death signal is triggered in the cell.  Antagonists of FasL-FasR interaction may include caspase inhibitors, mutated FasL/FasR, RNAi, or FasL/FasR antibodies. Agonists may include FasL/FasR encoding nucleotides.

Source: Exposing T cells to Fas Ligand (FasL)-Fas Receptor (FasR) Antagonists Withholds Differentiation and Increases Expansion Making T cells More Suitable for Use in Cancer Immunotherapy

Nitric Oxide-Based Therapeutics for Lung Cancer

The inventors have demonstrated a potent tumoristatic activity of JS-36-25, a diazeniumdiolate prodrug, in both lung cancer cells in vitro and as xenografts in mice.  JS-36-25 treatment led to 85% reduction of tumor growth in vivo.  The tumoristatic potency of the compound correlated well with the level of endogenous reactive oxygen species (ROS) in the cancer cells.  Thus, in addition to potent tumoristatic activity when administered alone, this compound is predicted to have a strong synergy with therapeutics that act through generation of ROS, such as bortezomib, doxorubicin, as well as high-energy radiation.

Source: Nitric Oxide-Based Therapeutics for Lung Cancer

Alpha-Glucosidase Chaperones and Inhibitors for Treatment of Pompe Disease and Type 2 Diabetes

Scientists at the NIH have discovered small molecules that can act as chaperones and correct the misfolding of mutated alpha-glucosidase enzyme.  Pompe disease is caused by deficiency or dysfunction of alpha-glucosidase.  The only FDA-approved treatment of Pompe disease is enzyme replacement, which is expensive and elicits an immune reaction in most patients that limits clinical utility.

NIH scientists also discovered small molecule inhibitors of alpha glucosidase enzyme.  Alpha glucosidase converts carbohydrates into monosaccharides.  Inhibition of this conversion is useful for type 2 diabetes.  Three FDA-approved inhibitors of alpha glucosidase exist but all have low efficacy:side effect ratios.

Source: Alpha-Glucosidase Chaperones and Inhibitors for Treatment of Pompe Disease and Type 2 Diabetes

Poly ADP-Ribose Polymerase Inhibitor/ Nitric Oxide Donor Dual Prodrugs as Anti-cancer Agents

Scientists at NIH have developed a hybrid prodrug molecule with enhanced biological activity as an anti-cancer agent. Novel cancer therapeutic strategies are in high demand. Diazeniumdiolate-based nitric oxide (NO)-releasing prodrugs are a growing class of promising anticancer agents.  Poly (ADP-ribose) polymerase (PARP) inhibitors have also emerged as another promising class of therapeutic compounds for cancer.  The two-component prodrug is expected to simultaneously deliver DNA damaging agent (NO release) with an inhibitor of DNA repair (PARP inhibitor) to a cancer cell.  The prodrugs are activated by glutathione/glutathione S-transferase (GSH/GST) and release cytotoxic NO and a PARP inhibitor in the target cancer cell.  The high levels of GSH/GST are often a feature of cancer cells.  The compound is predicted to have strong synergy with other anticancer therapeutics.

Source: Poly ADP-Ribose Polymerase Inhibitor/ Nitric Oxide Donor Dual Prodrugs as Anti-cancer Agents

Nitric Oxide-Releasing Polyvinylpyrrolidone-Based Polymers for Wound Healing and Related Applications

Novel nitric oxide-releasing polyvinylpyrrolidone-based polymers, their compositions, and use in treating wounds. The disclosed  polymers appear to be stable, biocompatible and bioabsorbable, while providing for extended nitric oxide release at therapeutic levels. The  invention also encompasses medical devices, such as wound dressings and bandages, which include the polymers and are capable of releasing nitric oxide when in use.  These devices may be used to treat a wound, various infections, and dermatological conditions.

The therapeutic efficacy of nitric oxide has been demonstrated for many indications, including wound healing.  As wounds are deficient in nitric oxide, its application has been shown to have beneficial effects on wound healing by promoting angiogenesis and tissue remodeling.

Source: Nitric Oxide-Releasing Polyvinylpyrrolidone-Based Polymers for Wound Healing and Related Applications

Diagnostic Test and Therapeutic Target for Sjogren's Syndrome

Sj?gren's syndrome is an autoimmune disease that attacks salivary glands resulting in chronic dry mouth and dry eyes.  Currently, there is no single diagnostic test to confirm the presence of Sj?gren's.  Physicians presently reach diagnosis after conducting a series of blood and functional tests for tear and salivary production. Diagnosis is further complicated as Sj?gren's symptoms frequently mimic those of other autoimmune diseases (e.g., lupus, rheumatoid arthritis, etc.) and is often overlooked as dryness associated with medications being taken by the patient.

Researchers at NIDCR have identified overexpression of a growth factor, bone morphogenetic protein 6 (BMP6), in patients with Sj?gren's.  By detecting BMP6 expression and/or activity, this invention potentially presents a singular confirmation to diagnose those suffering and those at risk for developing Sj?gren's.  BMP6 also presents a potential therapeutic target for Sj?gren's, a disease for which there is presently no cure.

Researchers have also discovered unique expression profiles for two other genes (XIST and MECP2) in male Sj?gren's patients.  Detecting aberrant expression and/or activity of these genes also offer a potential singular test for diagnosing Sj?gren's in male subjects.

Source: Diagnostic Test and Therapeutic Target for Sjogren's Syndrome

Axon Regeneration After Brain or Spinal Cord Injury

The invention is directed to modification of a particular sugar by the enzyme arylsulfatase B (ARSB), which results in axon regeneration.

Following traumatic brain or spinal cord injury, glial scars prevent regeneration of axons.  Chondroitin sulfate proteoglycans (CSPGs) are major components of glial scars.  CSPGs are made of a protein core containing glycosaminoglycan (GAG) sugar side chains, which, when sulfated, are responsible for the inhibitory activity of glial scars.  Specifically, NIH researchers have shown that the 4-sulfate unit on a certain sugar on GAG is responsible for inhibiting axon regrowth and, when the 4-sulfate unit is reduced, axon regrowth is observed.  Moreover, removal of this 4-sulfate unit by the ARSB enzyme promotes axon regrowth.

As a potential therapy for spinal cord injuries, researchers developed a vector expressing ARSB and demonstrated that this vector promotes axon regeneration when injected into the spinal cord of a mouse.

Source: Axon Regeneration After Brain or Spinal Cord Injury

Gag-Based DNA Vaccines Against HIV

Novel DNA vaccine constructs against HIV that express highly conserved elements (CE) within the HIV Gag protein and elicit strong, cross-clade cellular and humoral responses.  The DNA vaccine vectors were engineered to express CEs for protection against different clades of HIV and prevention of immunodominance, two issues associated with current HIV vaccine candidates.

In vivo studies of Rhesus macaques vaccinated with variants of these constructs expressing seven highly CEs covering >99 of all known Gag sequences elicited strong, cellular T-cell and humoral antibody immune responses.   The Gag-specific antibody responses were high titer and cross-clade.  Cross-clade protection is important given the sequence diversity of HIV as is the absence of immunodominant epitopes that generate antibodies which are not protective against HIV.

Source: Gag-Based DNA Vaccines Against HIV

Methods for Measuring Adrenomedullin and Monitoring and Treating Adrenomedullin-mediated Diseases, Such as Diabetes and Cancer

The technology includes methods for utilizing purified adrenomedullin (AM)-binding proteins, or functional portions thereof, to diagnose, treat, and monitor AM-related diseases such as diabetes and cancer. Antibodies and small-molecule antagonists, which can down-regulate the function of AM, Complement Factor-H (CFH), and the AM-CFH complex, have also been isolated.

AM is a ubiquitously-expressed peptide that functions as a universal autocrine growth factor. AM drives cell proliferation, acts as a vasodilator, can protect cells against oxidative stress in hypoxic injury, and acts as a dose-dependent inhibitor of insulin secretion. Methods for measuring in vivo levels of AM accurately and regulating the activity of available AM may be critically important in diagnosis and treatment of many conditions, such as heart disease, pulmonary disease, cirrhosis, cancer, diabetes, sepsis, and inflammation.

This technology centers on the observation that AM binds to CFH in vivo. Without a means to determine the amount of AM that is bound to CFH, measurements of AM are inaccurate. Furthermore, therapies focused on the AM-CFH complex may have advantages over therapies focused on AM alone.

Source: Methods for Measuring Adrenomedullin and Monitoring and Treating Adrenomedullin-mediated Diseases, Such as Diabetes and Cancer

Use of PAMP (proadrenomedullin N-terminal 20 peptide) and PAMP Inhibitors for the Treatment of Cancer, Cardiovascular Disease, and Other Angiogenesis-related Diseases

This technology details the use of PAMP or PAMP derivatives as a means to induce angiogenesis in tissue, as well as the use of PAMP inhibitors to inhibit angiogenesis.

PAMP (Proadrenomedullin N-terminal 20 peptide) is a 20 amino-acid molecule originating from the post-translational processing of pre-proadrenomedullin. PAMP is known as a potent hypotensive and vasodilatory agent; however, in addition to these properties, the inventors have discovered that PAMP also functions as a potent angiogenic factor. The inventors have also shown that an inhibitory fragment of PAMP, PAMP (12-20), is able to delay tumor growth in xenograft models of tumor progression.  The ability to promote angiogenesis can be used as a means to increase vascularization in specific tissue areas or to treat patients with ischemic diseases. In contrast, the ability to inhibit this process can be used to limit growth of solid tumors and as a therapy for retinopathies, endometriosis, or arthritis.

Source: Use of PAMP (proadrenomedullin N-terminal 20 peptide) and PAMP Inhibitors for the Treatment of Cancer, Cardiovascular Disease, and Other Angiogenesis-related Diseases

Topical Antibiotic with Immune Stimulating Oligodeoxynucleotide Molecules to Speed Wound Healing

The present technology provides a mean of improving the activity of topical antibiotics. Currently available topical antibiotic formulations effectively eliminate bacteria at a wound site.  But in eliminating bacteria in the wound, such antibiotics also eliminate the  molecular signals present in bacterial DNA that stimulate to immune system's wound healing processes.   Without these signals the rate of wound healing is diminished.  It would be desirable for topical antibiotics to remove infectious bacteria but also provide the immune stimulating signals needed to promote and accelerate healing.  The present formulation accomplishes these goals by supplementing the antibiotic formulation with immunostimulatory oligodeoxynucleotides (ODN).  These ODN express the CpG motifs present in bacterial DNA and safely mimic the immune stimulation induced by bacterial DNA.  The formulation may be applied directly to a wide variety of wounds to  skin (such as traumatic, burn, or surgical wound), or the eyes (such as corneal abrasions) to effectively eliminate infection and stimulate rapid healing of the wound.

Source: Topical Antibiotic with Immune Stimulating Oligodeoxynucleotide Molecules to Speed Wound Healing

Selective Treatment of Cancer, HIV, Other RNA Viruses and Genetically Related Diseases Using Therapeutic RNA Switches

Targeted therapy in cancer or viral infections is a challenge because the disease state manifests itself mainly through differences in the cell interior, for example in the form of the presence of a certain RNAs or proteins in the cytoplasm.

The technology consists of designed RNA switches that activate the RNA interference pathway only in the presence of a trigger RNA or DNA to which they bind, in order to knock down a chosen gene that is not necessarily related to the initial trigger.

This new approach can lead to a new type of drug that has the unique feature of selectively causing a biochemical effect (such as apoptosis) in cells that are infected by RNA viruses (such as HIV), as well as cancer cells. The RNA switch concept can be expanded to selectively treat other genetically related diseases.

Source: Selective Treatment of Cancer, HIV, Other RNA Viruses and Genetically Related Diseases Using Therapeutic RNA Switches

Activation of Therapeutic Functionalities with Chimeric RNA/DNA Nanoparticles for Treatment of Cancer, Viruses and Other Diseases

A new strategy based on RNA/DNA hybrid nanoparticles, which can be generally used for triggering multiple functionalities inside diseased cells is presented. Individually, each of the hybrids is functionally inactive and functional representation can only be activated by the re-association of at least two cognate hybrids simultaneously present in the same cell. Overall, this novel approach allows (i) the triggered release of therapeutic siRNAs or miRNAs inside the diseased cells, (ii) activation of other split functionalities (e.g. FRET, different aptamers, rybozymes, split proteins) intracellularly, (iii) higher control over targeting specificity (e.g. if two hybrids are decorated with two different tissue specific recognition moieties), (iv) biosensing and tracking of the delivery and re-association of these hybrids in real-time inside cells, (v) increasing the number of functionalities by introducing a branched hybrid structure, (vi) introduction of additional functionalities without direct interference of siRNA processivity, (vii) increasing the retention time in biological fluids by fine-tuning chemical stability through substituting the DNA strands with chemical analogs (e.g. LNA, PNA, etc.), (viii) conditional release of all functionalities.

Source: Activation of Therapeutic Functionalities with Chimeric RNA/DNA Nanoparticles for Treatment of Cancer, Viruses and Other Diseases

Treatment for Ichthyosiform Skin Diseases

A synthetic composition that contains the transglutaminase 1 (TGase I) enzyme and a lipid vesicle, which can be used to provide ameliorative therapy for inherited autosomal recessive ichthyoses (ARI). Icthyoses are rare inherited skin disorders that result in extensive scaling of the skin. Because this abnormality can affect heat and fluid transfer through the skin, individuals with this disease may have an increased risk for dehydration and skin infections. Each year, more than 16,000 babies are born with some form of ichthyosis. Ichthyosis affects people of all ages, races and gender. Currently, there is no cure for this disease and the only treatments available alleviate symptoms without affecting the disease itself. ARI are often caused by defects in lipid barrier function in the skin and are the result of genetic errors of either protein or lipid synthesis. One such disease, termed lamellar ichthyosis, is caused by genetic inactivation of the (TGase I) gene. The TGase I enzyme is essential for maintaining proper skin cornification, which protects skin cells against water loss and infection. Rather than simply treating the disease symptoms superficially, this technology provides a platform for treating the underlying cause of disease, namely the absence of TGase I function.

Source: Treatment for Ichthyosiform Skin Diseases

Treatment of Tuberculosis ? Adjuvant Therapies to Increase the Efficiency of Antibiotic Treatments

There is growing evidence that resistance to Mycobacterium tuberculosis infection is governed in large part by the regulation of host cell death.  Lipid mediators called eicosanoids are thought to play a central role in this process.  The subject invention is a novel method of enhancing the efficacy of antibiotic treatments for Mycobacterium tuberculosis infection by co-administering an inhibitor of 5-lipoxygenase and a COX-2 dependent prostaglandin.  Inhibition of 5-lipoxygenase and treatment with prostaglandin E2 results in alteration of the eicosanoid balance.   The synergistic effects of altering the eicosanoid balance and treatment with antibiotics is believed to result in more efficient reduction of the bacterial burden and thus, the period of antibiotic administration and antibiotic dosage could potentially be reduced.  In vivo data from mouse models can be provided upon request.

Source: Treatment of Tuberculosis ? Adjuvant Therapies to Increase the Efficiency of Antibiotic Treatments

A New Class, Now in Session: the First HLA Class II Restricted T Cell Receptor that Recognizes the Cancer Testis Antigen, MAGE-A3, Developed for Cancer Immunotherapy

NIH scientists have developed T cell receptors (TCRs) against the melanoma antigen family A3 (MAGE-A3) tumor antigen in the context of major histocompatibility complex (MHC) class II molecule HLA-DP-beta1*04. They are the first HLA class II restricted MAGE-A3 TCRs developed for use in adoptive immunotherapy. Previously developed MAGE-A3 TCRs are HLA class I restricted and generate CD8+ T cell responses to mediate tumor regression in some patients with MAGE-A3+ tumors. Other patients may not respond due to a lack of CD4+ T cells participation. Cancer immunotherapy with these new HLA class II TCRs could yield a robust and effective CD4+ T cell immune response that selectively targets MAGE-A3 expressing tumors without generating toxicity against healthy cells.

MAGE-A3 is a cancer testis antigen expressed on many types of cancer cells that blocks the functions of tumor suppressor proteins to mediate tumor growth and spreading. MAGE-A3 is not expressed on normal cells other than non-MHC expressing germ cells of the testis, which do not generate an immune response. Thus, MAGE-A3 represents an ideal target for cancer immunotherapies that are predicted to generate fewer toxic side effects than current standard cancer treatments.

Source: A New Class, Now in Session: the First HLA Class II Restricted T Cell Receptor that Recognizes the Cancer Testis Antigen, MAGE-A3, Developed for Cancer Immunotherapy

Novel Small Molecule Agonists of the Relaxin Receptor as Potential Therapy for Heart Failure and Fibrosis

The present invention is directed to novel small molecule agonists of the mammalian relaxin family receptor 1 (RXFP1), including human RXFP1.  Activation of RXFP1 induces: 1) vasodilation due to up-regulation of the endothelin system; 2) extracellular matrix remodeling; 3) moderation of inflammation by reducing levels of inflammatory cytokines; and 4) angiogenesis.  Small molecule agonists of RXFP1 may be useful in treating acute heart failure (AHF), scleroderma, fibrosis, other conditions associated with the biology of relaxin, and in improving reproductive health and wound healing.  These compounds are the first and only small molecule agonists of RXFP1.

Source: Novel Small Molecule Agonists of the Relaxin Receptor as Potential Therapy for Heart Failure and Fibrosis

Adeno-Associated Virus Gene Therapy for Diabetes and Obesity

This invention is directed to adeno-associated virus (AAV) vector delivery of exendin-4 (Ex-4) to salivary glands as treatment for diabetes and obesity. Ex-4 is a potent and long-acting agonist of the receptor for glucagon-like peptide 1 (GLP-1).  Scientists at NIDCR have shown that AAV-mediated delivery of Ex-4 resulted in improved glucose homeostasis and weight profile in two rat models of obesity and type 2 diabetes. Further, AAV-mediated delivery of Ex-4 to rat salivary glands resulted in localized and sustained expression of Ex-4 that was biologically active and well tolerated.

AAV-mediated delivery of Ex-4 is superior to administering GLP-1 analogs in that AAV-Ex-4 expression is more stable and longer acting. Like GLP-1 analogs, Ex-4 expression also potentially provides beneficial effects like reduced hypoglycemia, appetite suppression, and potential weight loss.

Source: Adeno-Associated Virus Gene Therapy for Diabetes and Obesity

Small Molecule MRS5474 with Anticonvulsant Activity for Treatment of Epilepsy

Adenosine modulates many physiological processes by activating specific adenosine receptors. These adenosine receptors play a critical role in the regulation of cellular signaling and are broadly distributed throughout the body. Thus, the ability to modulate adenosine receptor-mediated signaling is an attractive therapeutic strategy for a broad range of diseases. This technology relates to a group of compounds that display high affinity and specificity for the A1 adenosine receptor subtype.

One of the compounds, MRS5474, displays anticonvulsant activity in the 6 Hz animal model of clonic seizures. In the minimal behavioral toxicity test using the rotarod,  no toxicity (zero out of eight mice) was observed at all doses tested up to 30 mg/kg, the highest dose tested, which was nearly completely protective (seven out of eight animals) in the 6 Hz model. MRS 5474 also tested well in the corneal kindled mouse model to examine its effect on focal seizures.

Source: Small Molecule MRS5474 with Anticonvulsant Activity for Treatment of Epilepsy

Glucocorticoid-induced TNFR Family-Related Receptor Ligand (GITRL) Antibodies for Diagnosis and Treatment of Immune System Disorders

This technology provides novel antibodies and methods for diagnostics and treatment of disorders arising from dysregulation of the immune system using antibodies directed against glucocorticoid-induced tumor necrosis factor receptor family-related receptor ligand (GITRL). Also available are hybridomas producing anti-mouse GITRL monoclonal antibodies (clone 5F1).

Glucocorticoid-induced TNFR family-related receptor (GITR, also known as TNFRSF18) is expressed on the surface of responder T cells (CD4+CD25- or CD8+CD25- T cells).  Upon activation of the immune response, GITR is up-regulated and binds to its ligand, GITRL (also known as TNFSF18), which enhances the immune response.  The inventors have developed anti-GITRL monoclonal antibodies that block the interaction between GITR and GITRL, and have demonstrated in in vitro experiments that administration of these blocking antibodies can suppress the immune response. These antibodies may be useful for treatment of immune system disorders such as multiple sclerosis, rheumatoid arthritis, and other inflammatory diseases.

Source: Glucocorticoid-induced TNFR Family-Related Receptor Ligand (GITRL) Antibodies for Diagnosis and Treatment of Immune System Disorders

A Novel Immortalized Human Adrenal Cell Line with Inactive Protein Kinase A for Studies on cAMP Signaling and Endocrine Tumorigenesis

The first known immortalized cell line with a naturally-occurring inactivating mutation in PRKAR1A, the regulatory subunit type 1A (R1alpha) of protein kinase A (PKA), which is associated with tumor formation.

PKA isozyme balance is critical for the control of cAMP signaling and related cell cycle and proliferation changes. Aberrant cAMP signaling has been linked to adrenocortical and other, mostly endocrine, tumors. Inactivating mutations in the PRKAR1A gene are a known cause of Carney Complex - an autosomal dominant multiple neoplasia syndrome associated with skin, heart, and other myxomas and a variety of endocrine tumors.

Source: A Novel Immortalized Human Adrenal Cell Line with Inactive Protein Kinase A for Studies on cAMP Signaling and Endocrine Tumorigenesis

Cell Lines Expressing Nuclear and/or Mitochondrial RNase H1

RNase H1 has been shown to remove RNA/DNA hybrids and either too much or too little enzyme can lead to undesirable effects such as deletions of DNA. The gene encoding RNase H1 in mammalian cells produces two forms of the protein.  One is targeted to the nucleus of the cell and the other to the mitochondrial organelle. To study the effects of expression as well as to understand the regulation of the frequency with which each form is made, NIH investigators constructed cells derived from HEK293 cells where expression of each or both forms is/are expressed only after addition of doxycycline as a small molecule inducer compound. The set of cell lines could be important in the process of analysis of RNA/DNA hybrids as each cell line expresses different amounts of each form.

Source: Cell Lines Expressing Nuclear and/or Mitochondrial RNase H1

Cyclodextrins as Therapeutics for Lysosomal Storage Disorders

Cyclodextrins (CD), alone or in combination with other agents (e.g., vitamin E), as therapeutics for the treatment of lysosomal storage disorders (LSDs) caused by the accumulation of non-cholesterol lipids.

CDs are sugar molecules in a ring form. The alpha-CD (6 sugars), beta-CD (7 sugars) and gamma-CD (8 sugars) are commonly used cyclodextrins. The hydroxypropyl-beta cyclodextrin (HPbCD) has been approved for pharmaceutical use. Recent reports show that beta-cyclodextrin including HPbCD and beta-methyl-cyclodextrin reduced cholesterol accumulation and neuronal cell loss in the mouse model of NPC1 disease.

NCATS investigators found that CD (alpha-, beta- and gamma-CDs) increased intracellular Ca2+ and lysosomal exocytosis in both wild type cells and cells with Wolman disease, and reduced the size of enlarged lysosomes in six patient cell lines with LSDs. Further, CD in combination with tocopherol synergistically/additively reduced cholesterol accumulation in cells of NPC and Wolman diseases. Based on these results, they propose treatment of LSDs with cyclodextrins (such as alpha and gamma forms) alone or in combination with Vitamin E and its analogues for better efficacy and less side effects.

Source: Cyclodextrins as Therapeutics for Lysosomal Storage Disorders

Improved Transposase Compositions for Whole Genome Sequencing

The invention provides improved transposase enzymes engineered to exhibit reduced sequence biases, and to operate more efficiently than wildtype transposases.

Scientists at NIDDK and John Hopkins University jointly developed mutant transposases that are superior to wildtype transposases in whole genome sequencing applications.  Transposases facilitate the cleavage of certain DNA segments, called transposons, at specific sites within a genome and their subsequent insertions at random sites.  Addition of transposases and labeled transposons to whole genome preparations allow for one-pot, simultaneous fragmentation and identification of targeted DNA sequences.

Mutations introduced by the inventors facilitate formation of dimeric enzyme complexes with enhanced activity and stability.  These modifications result in more efficient fragmentation and tagging of genomic DNA.

Source: Improved Transposase Compositions for Whole Genome Sequencing

Improved Monoclonal Antibodies Against Neuregulin 2

The invention provides highly selective monoclonal antibodies against the extracellular domain (ECD) or intracellular domain (ICD) of neuregulin-2, a ligand for the ErbB receptors in adult human brain.  Neuregulins regulate a diverse array of neurological process in the central nervous system and are implicated in schizophrenia and other psychiatric disorders. However, an understanding of the specific role of neuregulin 2 has been hindered by a lack of specific antibodies useful in immunoblotting and immunohistology studies.  Commercially available antibodies do not perform as well in these applications when compared to the invention antibodies. A mouse monoclonal antibody directed to the ECD is available for licensing (clone 8D11, HHS Ref. No. E-192-2012), and rabbit antibodies directed to the ICD are also available (clone 11-11, HHS Ref. No. E-193-2012; clone 15-10, HHS Ref. No. E-189-2012; and clone 9-2, HHS Ref. No. E-188-2012). Antibodies from clones 8D11 and 11-11 have been validated for immunohistology and antibodies from clones 15-10 and 9-2 have been validated for Western blotting using brain tissue from wild-type and neuregulin 2 deficient mice.

Source: Improved Monoclonal Antibodies Against Neuregulin 2