Lina Cui

Demonstrates Good Selectivity and High Sensitivity for the Protease Within 10 Minutes

This fluorogenic probe rapidly detects cathepsin L activity with high sensitivity and selectivity. Cathepsin L is a lysosomal protease upregulated in many diseases and forms of cancer. Proteases are involved in the development and progression of cancer; inhibiting activity of cathepsin L may slow down cancer or disease progression.1 Direct measurement of the enzyme’s activity is necessary for developing pharmacologic inhibitors and assessing clinical samples for diagnostic purposes. However, available assays have various limitations. Standard fluorogenic probes have high background fluorescence that makes it difficult to ascertain true enzyme activity. These probes also can detect homologous enzymes such as cathepsin B and V by mistake, decreasing specificity to cathepsin L.

Researchers at the University of Florida have developed a fluorogenic probe that is highly selective to cathepsin L and not homologous enzymes. The probe’s structure results in less fluorescent background signal for better sensitivity to enzyme activity. Finally, activation of this fluorescent probe by cathepsin L is rapid, producing optimal results within 10 minutes.

Application

Fluorogenic probe for rapid, selective detection of cathepsin L enzymatic activity

Advantages

• Highly selective to cathepsin L activity, avoiding off-target detection of homologous enzymes in the cathepsin family
• Emits less fluorescent background signal, improving cathepsin L sensitivity for more accurate measurements
• Activates rapidly in the presence of cathepsin L, detecting enzymatic activity within 10 minutes

Technology

This fluorescent probe is chemically designed to favor the known substrate preferences of cathepsin L and not any of its homologous enzymes. Kinetic assays compared the responses of this probe and available probes to isolated cathepsin L, cathepsin B, and cathepsin V. The results indicate this newly synthesized probe has high selectivity to cathepsin L and low background fluorescence relative to available probes.

Single-Step Fluorescence Assay Facilitates High-Throughput Screening of Heparanase Inhibitors

This fluorogenic probe tests for the presence of heparanase and facilitates rapid screening for heparanase inhibitors to aid the development of cancer therapeutics. Heparanase is an enzyme overexpressed in many carcinomas, sarcomas, hematological malignancies, and inflammatory conditions. Upregulation of heparanase expression correlates with increased tumor size, metastasis, and poor prognosis. Detecting heparanase in the body may inform cancer therapies. The cancer diagnostics market should reach $250 billion by 2026. Common heparanase assays either lack sensitivity and/or selectivity, or require multiple steps with multiple reagents, or work at unfavorable conditions.

Researchers at the University of Florida have developed an ultrasensitive and selective heparanase small-molecule fluorogenic probe. This one-step assay is sensitive only to heparanase, and its derivatives developed in the same lab allows for use with medical imaging systems such as MRI or PET scans.

Application

Ultrasensitive heparanase small molecule fluorogenic probe to detect heparanase and screen for its inhibitors

Advantages

• Reliably detects heparanase, helping screen for it during the development of cancer therapeutics
• High sensitivity, accurately detecting even low levels of heparanase
• High selectivity, inhibiting interference from other similar biomolecules
• Assay provides simple, rapid, one-step heparanase fluorescence detection

Technology

The fluorogenic heparanase probe is comprised of two easily divisible parts, a recognition unit, and a fluorophore molecule linked by a glycosidic bond to the reducing end of the recognition unit. When heparanase breaks the glycosidic bond, the probe releases the fluorophore, which produces fluorescence. This fluorescence can be seen with a fluorescence microscope or a regular plate reader. The recognition unit is a disaccharide based on heparanase’s bond cleavage patterns. An electron-withdrawing group increases fluorescence, allowing the assay to indicate heparanase activity.

Contains Inorganic Nitrates and L-arginine to Facilitate Nitric Oxide Production

This mouthwash contains inorganic nitrates and L-arginine to facilitate nitric oxide (NO) production, enabling blood pressure reduction and regulation. Primary hypertension affects 34% of U.S. adults 20 years and older. While several factors, including demographic and environmental, are associated with primary hypertension, medical conditions such as periodontal diseases have recently been linked to elevated blood pressure levels.

Oral rinses are extensively used to prevent and treat caries, gingivitis, and periodontal disease. Oral antiseptic mouthwash and chlorhexidine gluconate (CHX) effectively reduce bacterial species associated with oral inflammation. However, the antibacterial effect of CHX is also associated with various detrimental effects on oral health, such as an increase in oral acidic condition, and, as a result, a change in saliva’s composition and the production of nitric oxide (NO), an important signaling molecule for regulating blood pressure and preserving vascular homeostasis. Consequently, using antiseptic mouthwash can negatively affect salivary and systemic NO levels, leading to increased blood pressure. This highlights a need for a novel oral hygiene product that enhances NO production, thereby enabling blood pressure reduction and regulation while supporting oral health. This is particularly important for patients with significant cardiovascular diseases who are hospitalized and have limited dexterity to exercise good oral hygiene.

Researchers at the University of Florida have developed a mouth rescue rinse containing nitric oxide (NO) precursors, such as inorganic nitrates and L-arginine, for blood pressure reduction and regulation while maintaining oral health. Featuring a novel composition, these precursors promote NO production. Rinsing with elevated levels of NO has proven to lower blood pressure in both normotensive and hypertensive individuals, preserving vascular homeostasis.

Application

Mouth rescue rinse containing nitric oxide precursors to reduce and regulate blood pressure while supporting oral health

Advantages

• Enhances oral nitric oxide production, reducing the cardiovascular risks associated with periodontal disease
• Contains L-arginine and inorganic nitrates, facilitating nitric oxide production for blood pressure regulation
• Comprises nitric oxide precursors, reducing blood pressure increases resulting from the use of antiseptic oral rinse by 50-90%
• Enables rinsing with elevated levels of nitric oxide, lowering blood pressure by 3-10 points in both normotensive and hypertensive individuals
• Provides a versatile formulation platform, supporting the development of multiple product formats, including toothpaste, gels, solutions, and tablets

Technology

This mouthwash facilitates blood pressure reduction and regulation through its novel composition. The formulation contains NO precursors, such as inorganic nitrates and L-arginine, supporting increased NO production. NO regulates blood pressure by relaxing and widening blood vessels, thereby reducing pressure within the arteries. Furthermore, when used before and/or after traditional chlorhexidine mouthwash, this formulation can eliminate the rise in blood pressure caused by the antiseptic oral rinse.