Nitric Oxide Services, LLC

10/09/2025

Thrilled to share our latest publication in Nitric Oxide:
“A Versatile Vanadium(III)-Based Chemiluminescence Protocol for Nitric Oxide Metabolite Quantification and NO Release Kinetics.”
🔗 https://pubmed.ncbi.nlm.nih.gov/41062015/

Nitric oxide (NO) is a critical signaling molecule at the intersection of cardiovascular, neurological, and immune health. Yet, its fleeting nature has long challenged efforts to measure it accurately in biological systems. Our work presents a robust, accessible, and cost-effective vanadium(III)-based chemiluminescence method that quantifies nitrate and nitrite — stable metabolites of NO — in biological fluids such as serum, urine, and cerebrospinal fluid.

By coupling this with headspace gas analysis, the method enables precise determination of NO release kinetics and half-life, allowing researchers and clinicians to better evaluate NO bioavailability and donor compound performance. The implications are broad — from monitoring endothelial function and sepsis progression to evaluating therapeutic efficacy and disease prevention strategies in personalized and translational medicine.

This work builds on decades of nitric oxide research and aims to make accurate NO metabolite analysis more routine across both diagnostic and research laboratories.
I extend sincere thanks to my co-authors Dr. Annette M. Sysel (Bauer Research Foundation, Inc.) and Dr. Michael J. Dunphy (Walsh UniversityWalsh University) for their collaboration and technical rigor.

Special appreciation to Alan and Ryan Kramme of Ace Glass Incorporated, Ken Thompson of Midwest Process Controls, and Dr. Daniel J. Smith of The University of Akron, whose foundational work inspired key refinements in this protocol.

Together, this represents a step forward toward making nitric oxide quantification more accessible — empowering researchers and clinicians to better prevent, monitor, and treat diseases rooted in NO imbalance.

Nitric oxide (NO) is a central mediator of vascular, neurological, and immune functions, and its dysregulation is implicated in a broad spectrum of diseases. Despite its significance, direct measurement of NO in human samples is limited by its transient nature and rapid conversion to nitrate and nit...

10/09/2025

Thrilled to share our latest publication in Nitric Oxide:

“A Versatile Vanadium(III)-Based Chemiluminescence Protocol for Nitric Oxide Metabolite Quantification and NO Release Kinetics.”

🔗 https://pubmed.ncbi.nlm.nih.gov/41062015/

Nitric oxide (NO) is a critical signaling molecule at the intersection of cardiovascular, neurological, and immune health. Yet, its fleeting nature has long challenged efforts to measure it accurately in biological systems. Our work presents a robust, accessible, and cost-effective vanadium(III)-based chemiluminescence method that quantifies nitrate and nitrite — stable metabolites of NO — in biological fluids such as serum, urine, and cerebrospinal fluid.

By coupling this with headspace gas analysis, the method enables precise determination of NO release kinetics and half-life, allowing researchers and clinicians to better evaluate NO bioavailability and donor compound performance. The implications are broad — from monitoring endothelial function and sepsis progression to evaluating therapeutic efficacy and disease prevention strategies in personalized and translational medicine.

This work builds on decades of nitric oxide research and aims to make accurate NO metabolite analysis more routine across both diagnostic and research laboratories.

I extend sincere thanks to my co-authors Dr. Annette M. Sysel (Bauer Research Foundation, Inc.) and Dr. Michael J. Dunphy (Walsh UniversityWalsh University) for their collaboration and technical rigor.

Special appreciation to Alan and Ryan Kramme of Ace Glass Incorporated, Ken Thompson of Midwest Process Controls, and Dr. Daniel J. Smith of The University of Akron, whose foundational work inspired key refinements in this protocol.

Together, this represents a step forward toward making nitric oxide quantification more accessible — empowering researchers and clinicians to better prevent, monitor, and treat diseases rooted in NO imbalance.

Nitric oxide (NO) is a central mediator of vascular, neurological, and immune functions, and its dysregulation is implicated in a broad spectrum of diseases. Despite its significance, direct measurement of NO in human samples is limited by its transient nature and rapid conversion to nitrate and nit...

10/07/2025

🚀 New Publication in Nitric Oxide (Pre-Proof) 🚀

I’m excited to share our latest paper from the combined efforts of Nitric Oxide Services, LLC, Bauer Research Foundation, Inc. and Walsh University:

“A Versatile Vanadium(III)-Based Chemiluminescence Protocol for Nitric Oxide Metabolite Quantification and NO Release Kinetics”

🔗 https://www.sciencedirect.com/science/article/abs/pii/S1089860325000874
@
This protocol finally provides researchers in the nitric oxide field with a true, step-by-step guide to the gold-standard NO analysis once limited to specialized NO laboratories.

✅ Standardized and ICH M10-aligned, this method is a game changer—a validated, cost-effective alternative to the widely used Griess assay.

✅ Enables reproducible quantification of NO metabolites in serum, urine, and cerebrospinal fluid.

✅ Supports direct measurement of NO release kinetics and compound half-life.

✅ Expands the role of nitric oxide as a biomarker in cardiology, neurology, oncology, and asthma control—empowering its use to guide clinical decision-making.

This standardized approach opens the door for broader clinical and translational adoption of NO metabolite analysis, uniting research and diagnostics under one reproducible platform.

08/29/2025

🌟 Publication Announcement 🌟

I’m excited to share that our new research has been published in Anticancer Research! https://ar.iiarjournals.org/content/45/9/3629

Our team including Annette M. Sysel, DVM, MS, Michael Dunphy, Ph.D. and myself... investigated SCC5, a silver-based compound, as a potential alternative to traditional platinum chemotherapy drugs (such as cisplatin). Platinum drugs have saved countless lives, but they are often limited by serious side effects. In contrast, SCC5 showed:

✅ Strong activity against several types of cancer cells (melanoma, colon, ovarian, prostate, and more)

✅ A significantly better safety profile in preclinical testing

✅ Evidence of triggering natural cell death pathways in cancer cells

This work highlights how silver, long known for its healing properties, may play an important role in the future of cancer therapy.

💡 A huge thank you to the reviewers and editorial team for their thoughtful insights, and to the institutions that made this work possible:

Nitric Oxide Services, LLC (Akron, OH)

Bauer Research Foundation(Akron, OH)

Walsh University (North Canton, OH)

And a special tribute to The University of Akron especially Dr. Wiley J. Youngs, whose pioneering work in silver carbene chemistry paved the way for this study.

We are only at the beginning, but this is a promising step forward in developing safer, more effective cancer treatments.

🔗 Read more here: https://ar.iiarjournals.org/content/45/9/3629

Background/Aim: Platinum-based chemotherapeutics remain the standard of care for many solid tumors; however, they have dose-limiting toxicities. Silver(I) N -heterocyclic carbene complexes represent a novel class of metallodrugs with potential anticancer activity and improved tolerability. This stud...

07/14/2025

Congratulations to Dr. Brian Leyshon - Abbott- and Team!

Thrilled to see the late-breaking abstract presented at The European Society for Clinical Nutrition and Metabolism (ESPEN) - : “A Novel Lipid Technology Improves Vitamin D3 Absorption in Healthy Adults”

This innovative study—led by Dr. Brian Leyshon and colleagues from Abbott Laboratories, Walsh University, Nitric Oxide Services, LLC, and the University of Dayton—demonstrates that a new lipid-based formulation (MDGPL) significantly enhances the absorption of vitamin D3 in healthy adults. Notably, the MDGPL technology improved absorption by up to 54% at standard dietary doses, leading to meaningful increases in serum 25(OH)D3 levels.

Given that vitamin D insufficiency remains a global challenge, these findings offer promising solutions for improving nutritional status and health outcomes.

Proud to be part of this collaborative effort and excited to see the impact of these advances on clinical nutrition!

Presented at: ESPEN 2025, Late-Breaking Abstract Session

Authors: Brian J. Leyshon, Steven R. Hertzler, Kelsey F. Scanlon, Michael J. Dunphy, Qi Yao, Joseph A. Bauer, Joseph I. Harper, Lindsay K. Schultz



Link to ESPEN Programme & Abstract:

Explore the ESPEN 2025 Congress Main Programme for a comprehensive overview of cutting-edge clinical nutrition and metabolism sessions.

07/05/2025

🚀 New Publication Alert! 🚀

Looking back on 25+ years since earning my doctorate in biochemistry, if I had to pick one assay that has truly defined my career, it would be the Sulforhodamine B (SRB) assay. This robust, versatile method verified the anticancer activity of a compound I synthesized—nitrosylcobalamin—and laid the foundation for a landmark publication in the JNCI: Journal of the National Cancer Institute.

The SRB assay has been at the core of my teaching and mentoring, helping train countless students at both university and high school levels. It has allowed me to share the excitement of anticancer research with students, educators, and colleagues alike. Special recognition to Dr. Amy Heston, who trained several outstanding students at Walsh University, leading to published studies on the effects of copper and heavy metals on various human cancer cell lines.

I'm proud to share that we have now published a detailed, step-by-step guide to the SRB assay, making it accessible to educators and research labs everywhere. Our hope is that this protocol will empower more teams to screen both natural and synthetic compounds for anticancer activity, expanding the arsenal in the fight against cancer.

Check out our new publication here:
🔗 ScienceDirect: A Versatile Vanadium(III)-Based Chemiluminescence Protocol for Nitric Oxide Metabolite Quantification and NO Release Kinetics https://www.sciencedirect.com/science/article/pii/S2215016125003140?via%3Dihub

Let’s inspire the next generation of cancer researchers and educators—one assay at a time!

04/01/2025

Welcome to Nitric Oxide Services, LLC (NOS) — Your CLIA-Certified Partner in Nitric Oxide Science

Nitric Oxide Services, LLC (NOS) is a premier consulting and research organization dedicated to advancing the science and application of nitric oxide and related biochemical pathways in health, medicine, and exercise science. Our mission is to help clients optimize physiological function, improve clinical outcomes, and drive innovation through nitric oxide-based solutions.

As a CLIA-certified laboratory (CLIA # 36D2321191), we meet the highest federal standards for laboratory quality and compliance. This certification enhances our ability to deliver validated, high-precision nitric oxide (NOx: nitrate and nitrite) biomarker testing, supporting both clinical trials and product development with data you can trust.

Backed by a multidisciplinary team of seasoned researchers, clinicians, and consultants with decades of combined experience, NOS operates at the forefront of nitric oxide science and translational research.

Our laboratory is equipped with:

State-of-the-art analytical instrumentation

Tissue culture facilities

Specialized chemical reactors

CLIA-compliant biomarker testing services

In addition, we have privileged access to the Comparative Medicine Unit (CMU) — a 41,000-square-foot, fully accredited preclinical research facility. The CMU maintains full compliance with USDA regulations and holds an Animal Welfare Assurance from the Office of Laboratory Animal Welfare (OLAW), supporting advanced preclinical studies.

At NOS, we integrate cutting-edge research, regulatory expertise, and clinical trial support to advance the boundaries of nitric oxide science. Whether you are developing medical devices, supplements, or therapeutic interventions, we are here to accelerate your innovation with trusted data and proven results.

At Nitric Oxide Services, we specialize in comprehensive validation of medical devices, supplements, nitric oxide precursors and donors especially within the realm of applications in health and medicine including exercise science. We have worked successfully to advance client projects in the areas o...

04/01/2025

📣 We are officially CLIA-Certified!

Nitric Oxide Services, LLC has been awarded a CLIA Certificate (CLIA # 36D2321191) — a critical milestone that underscores our commitment to delivering high-quality, compliant, and reliable laboratory testing services.

Why does this matter for you?

✔️ CLIA certification ensures that our laboratory meets rigorous federal quality standards for accuracy, reliability, and timeliness of results — giving you confidence in the data that supports your clinical trial.

✔️ We specialize in nitrate, nitrite (NOx), and nitric oxide-based biomarker testing — essential endpoints for trials focused on cardiovascular health, exercise physiology, metabolism, and beyond.

✔️ Our team is ready to support sponsors, CROs, and investigators with customized testing solutions designed to strengthen your research outcomes.

Whether you are launching a new clinical trial or enhancing an existing one, we are here to provide validated, regulatory-compliant assays to meet your needs.

🔗 Contact us today to learn how Nitric Oxide Services, LLC can bring added value to your clinical research.

https://www.nitricoxidelab.com/

At Nitric Oxide Services, we specialize in comprehensive validation of medical devices, supplements, nitric oxide precursors and donors especially within the realm of applications in health and medicine including exercise science. We have worked successfully to advance client projects in the areas o...

04/01/2025

⚖️ FDA’s LDT Rule Struck Down — What Happens Next for Diagnostic Innovation?

The FDA recently lost its attempt to regulate Lab-Developed Tests (LDTs) as medical devices after a federal judge vacated the rule.

For laboratories which already validate tests under ICH M10 and meet CLIA requirements, this ruling highlights a deeper issue:

Are we adding regulations for the sake of control, or are we solving a real problem?

In this blog, I offer a balanced view of: ✅ Why the rule was made
✅ Why the court rejected it
✅ The future of LDTs, innovation, and patient access

This moment matters — for labs, clinicians, and most importantly, patients.

https://thebiochemistknowsbest.com/f/fdas-now-vacated-rule-on-ldts-unnecessary-fix

FDA's Now-Vacated Rule on LDTs: Unnecessary Fix for a Problem That Didn’t Exist?

03/28/2025

Over 25 years ago, I synthesized and patented a molecule called Nitrosylcobalamin (NO-Cbl)—a vitamin B12 analog designed to deliver nitric oxide (NO) therapeutically. Inspired by groundbreaking work by Brouwer et al. (Blood, 1996) and Greenberg et al. (JPET, 1995), who observed natural formation of NO-cobalamin interactions in the body, I sought to create a stable and bioactive compound that harnessed these biochemical interactions.

Today, I'm humbled to see over 20 peer-reviewed studies on NO-Cbl listed in PubMed, the latest of which was just published:

https://pubmed.ncbi.nlm.nih.gov/40150986/

Yet, the journey hasn't been easy. Nitrosylcobalamin sparked controversy among traditional vitamin B12 researchers, especially when I collaborated with renowned B12 expert Dr. Bellur Seetharam. Instead of welcoming innovative approaches, some sought to undermine this new direction—even pushing unsuccessfully to rename the molecule inaccurately to "nitroxylcobalamin."

I've often thought of sharing these behind-the-scenes stories in academia, where collaboration sometimes gives way to rivalry and politics. My experience is not unique; many researchers face similar resistance when challenging established paradigms.

However, seeing my work validated and expanded upon by other researchers gives me renewed energy and hope. I'll continue working diligently to translate these findings into clinical breakthroughs to ultimately improve patient outcomes.

To fellow innovators facing similar battles: keep pushing forward—the science, and those who will benefit from it, depend on your perseverance.

Variations in IF binding between NO-Cbl and OH-Cbl may be attributed to the saturation of binding sites or recognition issues specific to plant-derived IF. This study underscores the potential of NO-Cbl as a targeted therapeutic agent capable of leveraging natural cobalamin uptake pathways. These re...

03/18/2025

We're excited to announce the publication of our latest research study in Frontiers in Bioscience (Elite Edition)!

Our study, titled "Evaluation of the Binding Affinity of Nitrosylcobalamin to Intrinsic Factor as a Predictive Model for Cobalamin Binding Protein Interactions: A Comparative Study with Hydroxocobalamin", explores the binding dynamics of nitrosylcobalamin (NO-Cbl), a promising vitamin B12 analog with potential therapeutic applications, especially in cancer treatment.

🔬 Key Highlights:

We utilized Surface Plasmon Resonance (SPR) to analyze the binding affinity of NO-Cbl and hydroxocobalamin (OH-Cbl) to recombinant human intrinsic factor (IF) derived from plants.
Both compounds demonstrated high binding affinities in the picomolar range, showcasing their potential for targeted therapeutic delivery.

Our data revealed nuanced differences in binding stability and kinetics, important considerations for developing targeted therapeutics leveraging natural cobalamin uptake pathways.

🌱 This work underscores the effectiveness of plant-derived IF as a reliable and scalable model for predicting the biological interactions of cobalamin analogs.

📖 Read the full open-access article here: https://www.imrpress.com/journal/FBE/17/1/10.31083/FBE26810

We're grateful for the collaborative efforts and insightful reviews that helped strengthen this study. Special thanks to our colleagues at Cleveland Clinic and Cobento Biotech for their invaluable contributions.

Let's connect and discuss how NO-based therapies can revolutionize targeted treatment strategies!

Background: Intrinsic factor (IF) is a glycoprotein crucial for cobalamin (vitamin B12) absorption in the human body. This study aimed to evaluate the binding affinity of nitrosylcobalamin (NO-Cbl), a cobalamin analog, to recombinant human IF derived from plants, using hydroxocobalamin (OH-Cbl) as a...