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OUR STUDIES

Functional Assurance and Evidence-based Results

Potency, or bio-functional status, of an active principle is dependent on molecular separation (isolation) and structural stability of a bioactive compound. Protein engineering to map the three-dimensional structures of bioactive molecules is a specialized field. bioQuad® Life Sciences is a leading world expert in protein engineering. The first activation of any protein structure outside an in vivo system was first reported by Dr. Naidu and his team in the year 2000 with ‘Activated Lactoferrin’ – a research finding that made global headlines. The purity (certificate of analysis) and homogeneity of a biochemical does not necessarily assure the potency of such molecule. Bioactive molecules ‘unfold and die’; therefore, their stability data and expiry date (best before use) is disclosed for nutritional ingredients. Our laboratory routinely performs biological testing of active ingredients and their finished products to ensure their functional performance. ‘Quality Assurance’ is an important GMP protocol, but we also believe that ‘Functional Assurance’ (which is not a standard norm practiced in the health supplement industry) is much more critical to ensure the biological performance (potency) of the product.

Lactoferrin Treated for Contaminant Reduction (LF-TCR)

patented 3 tier filtration process of milk lactoferrin

Lactoferrin is a milk protein credited with an impressive list of health benefits. Separation of lactoferrin from other ingredients of milk requires several complex steps of protein engineering. Despite high purity, proteins isolated from such processes may harbor microbial and endotoxin contaminants that could compromise lactoferrin functionality. A novel treatment for contaminant reduction (TCR) to enhance the protein quality during commercial-scale lactoferrin production has been developed. Lactoferrin / LF-(TCR), based on this cutting-edge protein technology, contains ultra-clean lactoferrin that could offer the highest standards of microbiological quality and functional assurance for nutraceutical applications.

Intestinal Maintenance

Background - The human intestinal tract (consisting of small and large intestines) is a long (30') semi-permeable biological tube that separates the true inside of the body from the outside. Its major functions include to i) absorb digested food (nutrients, energy, water), ii) excrete waste material (feces) and iii) keep out pathogens (serve as a defense barrier). The lining of the intestine is covered with microvilli (tiny projections) that exponentially increase the surface area of the intestinal mucosa. The integrity and density of microvilli is vital for optimal absorption of nutrients.

human body showing intestinal lining

Study Objective - To preserve the integrity and density of the intestinal microvilli, which is vital for efficient nutrient absorption.

intestinal cell lining preserved by lactoferrin TCR

Study Results - In this CaCO2 cell culture experiment, intestinal epithelia cells were grown as monolayers in vitro. The monolayers were visualized under phase-contrast microscopy for integrity and morphology. Normal viable cell monolayer appear confluent, compact, round and bright. We observed the cell morphology over 24 hours. There were four cell culture groups - CONTROL (cells without any treatment); and cells treated with i) whey LF, ii) milk LF and iii) milk LF-(TCR). After 24 hours, as shown - the CONTROLS showed cell death (apoptosis with dark brown shade). Monolayers treated with whey and milk LF actually showed both apoptosis and cytotoxicity resulting from protein exposure (dark murky color). In contrast, monolayers treated with LF-(TCR) showed confluent cell growth with healthy structural morphology (bright color with no discoloration).

Reference: Naidu AS (2005) Ultra-cleansing of lactoferrin: Nutraceutical implications. Eur J Nutraceut Funct Foods 16 (March/April):7-13.

Host Defense

Background - The human immune system has three lines of host defense. The 1st line (or surface ‘frontline’) of defense consists of non-specific physical and chemical barriers that readily defend the body. The 2nd line of defense is a non-specific immune response that involves macrophages, cytokines, interferons, neutrophils and complement cascade. The 3rd line of defense is specific resistance that relies on specific antibodies produced by the body to recognize, fight and eliminate foreign microbes. The interplay between these three lines of host defense supports a healthy immune system.

immune cells engulfing microbes

Study Objective - To compare the effect of various lactoferrin preparations on bacterial growth-multiplication.

Study Results - In this experiment, we compared the effect of different LF preparations on growth-multiplication of bacteria. Green-fluorescent protein (GFP)-labelled bacteria were exposed to 1% LF containing broth solutions for 24 h. Broth without any LF served as positive growth CONTROL. The intensity of GFP (measured by florescent microscopic imaging) reflected the extent of bacterial growth-multiplication. Thus, higher the bacterial growth-multiplication, brighter the green color as visualized in the CONTROL broth (without LF). Whey LF and milk LF showed a slight decrease in the green fluorescence. In contrast, the LF-(TCR) has inhibited the growth-multiplication of the bacteria - resulting in no enhancement of the green fluorescence. 

Reference: Naidu AS (2005) Ultra-cleansing of lactoferrin: Nutraceutical implications. Eur J Nutraceut Funct Foods 16 (March/April):7-13.

Ribonuclease-Enriched Lactoferrin (RELF)

Bone Turnover

Background - The bone is a dynamic tissue that undergoes a continuous remodeling process through repeated cycles of destruction and rebuilding. In healthy adults, the amount of new bone formation approximately balances the amount of bone resorption. As age increases, and due to factors such as diet, lifestyle, medications, etc., the balance shifts to favor bone resorption. A reduction in angiogenesis could also contribute to this imbalance. Maintaining healthy bone turnover is essential to reinforce skeletal structure and optimize its function.

hip bone with bone matrix background

Study Objective - Examine the effect of RNase-enriched lactoferrin (R-ELF) supplement on healthy bone turnover

bone resorption decreased and bone formation increased by ribonuclease-enriched lactoferrin

Study Results - A total of 35 healthy, post-menopausal women, aged 45 to 60 years, were randomized into placebo or R-ELF supplement group. Over a six-month period, the bone status was monitored by assessing bone resorption markers, serum N-telopeptides (sNTx) and urine deoxypyridinoline (uDpd) cross-links; and serum bone formation markers, bone-specific alkaline phosphatase (SBAP) and osteocalcin (sOC). R-ELF supplmentation demonstrated a decrease in uDpD levels by 14% (19% increase in placebo) and sNTx maintained at 25% of the baseline (41% for placebo), while serum BAP and OC levels showed a 45% and 16% elevation (25% and 5% for placebo). R-ELF supplementation showed positive effects on healthy bone turnover.

Reference: Bharadwaj S, Tezus Naidu AG, Betagiri GV, Prasadarao NV, Naidu AS (2009) Milk ribonuclease-enriched lactoferrin induces positive effects on bone turnover markers. Osteo Int 20(9):1603-1611.

Joint Health

Background - Bone and joints work together to enable movement, similar to a joist and lever system. For optimal function, the joints have to preserve structure, maintain lubrication (i.e. synovial fluid) and remove debris (normal by-product of wear-and-tear). The body relies on the immune system to protect the joints by continuously monitoring its status; when there is excessive wear-and-tear, the immune system triggers the release of several cytokines to indicate a 'high alert' state. Optimal joint health can only be achieved when these processes are addressed and in balance.

human body showing joint inflammation process

Study Objective - Evaluate the effect of RNase-enriched lactoferrin (R-ELF) supplement on joint health (by measuring circulating cytokine levels).

pro-inflammation decreased and anti-inflammation increased by ribonuclease-enriched lactoferrin

Study Results - A six-month, randomized clinical study was conducted on 35 healthy, post-menopausal women, aged 45 to 60 years. They were randomized into placebo or R-ELF supplement group; the R-ELF group was supplemented with R-ELF (2x 125 mg/day) and calcium (100% RDA), while the placebo group received only the calcium supplement. Serum levels of receptor activator for NF-κB ligand (RANKL), C-reactive protein (CRP) and various other cytokines were determined by ELISA. IL-6 and TNF-α decreased significantly (-44% and -10% respectively) while IL-10 increased (140%) with R-ELF supplementation. RANKL and CRP were modestly reduced (-50%) relative to the placebo, while RANKL elevated initially. R-ELF supplementation showed beneficial effects on joint health.

Reference: Bharadwaj S, Tezus Naidu AG, Betagiri GV, Prasadarao NV, Naidu AS (2010) Milk ribonuclease-enriched lactoferrin supplementation in women. J Infl Res 59(11):971-978.

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