Immunomodulatory Effects of Modified Bovine Colostrum, Whey, and Their Combination with Other Natural Products: Effects on Human Peripheral Blood Mononuclear Cells.

Background: Many natural products have immunomodulatory properties. However, the mechanism of immunomodulatory activities are poorly understood. Objectives: This study evaluated the influence of bovine colostrum products, a whey product, or their combinations with other natural products on human peripheral blood mononuclear cells' (PBMC) ability to produce cytokines upon activation. Methods: PBMCs were pretreated with ultrafiltered colostrum, nano-filtered bovine colostrum, egg yolk extract, a botanical blend, colostrum + egg yolk extract, colostrum + egg yolk + botanical blend, and fermented whey and then stimulated with lipopolysaccharide or phytohemagglutinin. Cytokine production was measured by the Luminex assay. Results: All study products demonstrated immunomodulatory properties by regulating cytokines production by activated PBMCs. Ultrafiltered colostrum alone displayed the highest immune stimulatory activity. It stimulated proinflammatory cytokine production by lipopolysaccharide-activated PBMCs and suppressed cytokine production by phytohemagglutinin-activated cells. Other study products mainly suppressed cytokine release by both cell types. The immunomodulatory properties depended upon the dose of the products used in the study. Conclusions: All tested products modulated innate and adaptive immune cell activities. Most of the products demonstrated anti-inflammatory properties, except ultrafiltered colostrum, which stimulated the lipopolysaccharide-activated PBMC production of inflammatory cytokines. These products can be potentially used to support overall immune health.

In Vitro Assessment of Gold Nanoparticles on Telomerase Activity and Telomere Length in Human Fibroblasts.

Telomerase activity coincides with lengthening of the ends of chromosomes known as telomeres. Telomere length is used as a marker for cellular aging. Telomeres shorten over time as cells divide, and certain bioactive compounds such as gold nanoparticles (AuNPs) may slow the shortening of telomeres by increasing telomerase activity. The objective of the present study is to assess the effect of AuNPs on telomerase activity and telomere length in human fibroblasts. Telomerase activity was measured using enzyme-linked immunosorbent assay (ELISA) in primary human lung fibroblasts (IMR90) and using quantitative PCR-based telomeric repeat amplification protocol (Q-TRAP) in primary human dermal fibroblasts, neonatal (HDFn). Telomere length was determined by Telomere Analysis Technology (TAT®)assay in HDFn. In IMR90, all AuNP treatments showed significant increases in telomerase activity when compared to earlier passages. HDFn treated with AuNPs at 0 ppm, 0.05 ppm, 0.5 ppm, or 5 ppm did not show significant differences in telomerase activity compared to the control group. Significant differences in telomere length in HDFn were observed at 2 weeks of 0.05 and 0.5 ppm AuNPs under oxidative culture conditions as compared to the control group. The study showed preliminary evidence that AuNPs may increase telomerase activity and decelerate the shortening of telomeres in human fibroblasts, suggesting its potential anti-aging effects, which warrants further investigation.

Safety and Efficacy of a Multivitamin, Multimineral, Bovine Colostrum-Containing Supplement: An Open-label Pilot Intervention Trial in Healthy Adult Women and Men.

Objective: This study was undertaken to explore the effects of a bovine colostrum-containing multivitamin multimineral (MVM) supplement on healthy, adult women and men by determining blood chemistries and health parameters via serum and saliva sampling and measuring each subject's physical characteristics over a 12-week interval. Participants: Fifty participants were screened for the study, after which twenty participants were determined eligible to enter the study. Thirteen participants (6 women and 7 men, average age 30.9 years, average BMI 27.3 kg/m2) completed the whole study. Results: MVM did not significantly impact serum alanine aminotransferase (ALT), aspartate aminotransferase (AST), gamma-glutamyl transpeptidase (GGT) levels. The MVM significantly improved serum folate levels (48.3% increase at Week 12: 23.56 ± 5.75 ng/mL versus Week 0: 15.88 ± 3.40 ng/mL, P = .0001). MVM improved serum levels of vitamin B12 (21.3% increase at Week 12: 789.38 ± 313.23 pg/mL versus Week 0: 650.54 ± 228.02 pg/mL, P = .0690) and 25-hydroxy-vitamin D levels (9.1% increase at Week 12: 32.22 ± 5.81 ng/mL versus Week 0: 29.54 ± 11.30 ng/mL, P = .3570). The salivary IgA levels showed a significant increase at Week 4 (249.85 ± 95.63 ng/ml), Week 8 (271.65 ± 133.52 ng/ml), and Week 12 (279.88 ± 128.19 ng/ml) compared to Week 0 (177.57 ± 74.81 ng/ml). Conclusions: This study shows that MVM has a good safety and tolerability profile and can be used as a daily nutritional supplement safely. MVM may improve serum levels of vitamin D, folate, vitamin B12, and, possibly, other blood markers. The study showed that MVM may improve secretory IgA levels, a major component of oral immunity. These findings suggest an overall improvement in several aspects of health and need to be confirmed in a larger, placebo-controlled study.

A Clinical Study to Evaluate the Safety and Efficacy of Oral Administration of Microscopic Dose Gold Nanoparticle (AuNP) on Knee Joint Health and Function in Arthritis Patients.

The purpose of this clinical study was to determine whether gold nanoparticle (AuNP) supplementation at a dosage of 0.34 mg elemental gold per day can improve knee joint health, function, and quality of life for arthritis patients. A total of 51 participants (24 male and 27 female, age 62.1 ± 13.1) were followed for 20 weeks through a three-phase longitudinal study. Both subjective and objective parameters were used to measure changes in joint health and function, as well as quality of life. The study found patients' Knee injury and Osteoarthritis Outcome Score (KOOS) improved with statistical significance. It was reported that 71.42% of the cohort experienced improvements in their perceived knee pain and 61.22% with improvements in knee stiffness. Majority of objective measurements such as pain with range of motion and specific exercises requiring proper knee health and function did not show statistically significant improvement but did show a positive improving trend in support of AuNP supplement. Study cohort showed statistically significant improvements in two specific exercises: sit-to-stand and single-leg squat. By the end of the study, 70% of the study cohort indicated that they would continue to take the supplement even after the study concluded. Though the study has limitations and is not definitely conclusive, it was the first clinical study to show that oral micro-dosage of AuNP as low as 0.34 mg daily is safe and effective for both rheumatoid arthritis and osteoarthritis patients. This study opened way for the use of AuNP in both clinical and daily settings to improve joint health and function for both average and athletic users.

Influence of 3D Printing Parameters on the Mechanical Stability of PCL Scaffolds and the Proliferation Behavior of Bone Cells.

Introduction The use of scaffolds in tissue engineering is becoming increasingly important as solutions need to be found for the problem of preserving human tissue, such as bone or cartilage. In this work, scaffolds were printed from the biomaterial known as polycaprolactone (PCL) on a 3D Bioplotter. Both the external and internal geometry were varied to investigate their influence on mechanical stability and biocompatibility. Materials and Methods: An Envisiontec 3D Bioplotter was used to fabricate the scaffolds. First, square scaffolds were printed with variations in the strand width and strand spacing. Then, the filling structure was varied: either lines, waves, and honeycombs were used. This was followed by variation in the outer shape, produced as either a square, hexagon, octagon, or circle. Finally, the internal and external geometry was varied. To improve interaction with the cells, the printed PCL scaffolds were coated with type-I collagen. MG-63 cells were then cultured on the scaffolds and various tests were performed to investigate the biocompatibility of the scaffolds. Results: With increasing strand thickness and strand spacing, the compressive strengths decreased from 86.18 + 2.34 MPa (200 µm) to 46.38 + 0.52 MPa (600 µm). The circle was the outer shape with the highest compressive strength of 76.07 + 1.49 MPa, compared to the octagon, which had the lowest value of 52.96 ± 0.98 MPa. Varying the external shape (toward roundness) geometry, as well as the filling configuration, resulted in the highest values of compressive strength for the round specimens with honeycomb filling, which had a value of 91.4 + 1.4 MPa. In the biocompatibility tests, the round specimens with honeycomb filling also showed the highest cell count per mm2, with 1591 ± 239 live cells/mm2 after 10 days and the highest value in cell proliferation, but with minimal cytotoxic effects (9.19 ± 2.47% after 3 days).

Method development for the analysis of PBMC-mediated killing of K562 cells by bovine colostrum.

BACKGROUND: Colostrum is the first milk that supplies newborns with immune supporting peptides. Due to its heterogeneous and variable characteristics, standardized assays for assessment of its biological activities are a challenge. The current set of studies were aimed to investigate the immune activity of bovine colostrum blends as well as develop a method to assess variability across different lots. METHODS: Immune activity of a bovine colostrum blend was evaluated by their ability to enhance PBMC (peripheral blood mononuclear cell)-mediated killing of K562 cells. K562 cell killing was assessed by flow cytometry using DAPI. PBMCs derived from multiple healthy donors were initially investigated. Frozen PBMC aliquots from one of the highest responders were used for subsequent studies. Different doses and lots of product were assessed. Incubation time was also explored. RESULTS: Bovine colostrum blend similarly reduced K562 cells number and these effects were often greater than the IL-2 positive control. Despite consistent efficacy at enhancing PBMCs-mediated K562 killing, the degree of the effect was significantly variable across different lots. These biological effects were largely dependent on the solubility of the product. CONCLUSION: Assessment of PBMC-mediated killing of K562 cells by flow cytometry using DAPI can be a reliable method for measuring immune activity of bovine colostrum when the material is well-dissolved into solution and the same biological sample from a single donor is used.

Enhancing Skin Health: By Oral Administration of Natural Compounds and Minerals with Implications to the Dermal Microbiome.

The history of cosmetics goes back to early Egyptian times for hygiene and health benefits while the history of topical applications that provide a medicinal treatment to combat dermal aging is relatively new. For example, the term cosmeceutical was first coined by Albert Kligman in 1984 to describe topical products that afford both cosmetic and therapeutic benefits. However, beauty comes from the inside. Therefore, for some time scientists have considered how nutrition reflects healthy skin and the aging process. The more recent link between nutrition and skin aging began in earnest around the year 2000 with the demonstrated increase in peer-reviewed scientific journal reports on this topic that included biochemical and molecular mechanisms of action. Thus, the application of: (a) topical administration from outside into the skin and (b) inside by oral consumption of nutritionals to the outer skin layers is now common place and many journal reports exhibit significant improvement for both on a variety of dermal parameters. Therefore, this review covers, where applicable, the history, chemical structure, and sources such as biological and biomedical properties in the skin along with animal and clinical data on the oral applications of: (a) collagen, (b) ceramide, (c) ß-carotene, (d) astaxanthin, (e) coenzyme Q10, (f) colostrum, (g) zinc, and (h) selenium in their mode of action or function in improving dermal health by various quantified endpoints. Lastly, the importance of the human skin microbiome is briefly discussed in reference to the genomics, measurement, and factors influencing its expression and how it may alter the immune system, various dermal disorders, and potentially be involved in chemoprevention.

Thermogenic Blend Alone or in Combination with Whey Protein Supplement Stimulates Fat Metabolism and Improves Body Composition in Mice.

BACKGROUND: Certain food ingredients promote thermogenesis and fat loss. Similarly, whey protein improves body composition. Due to this potential synergistic effect, a blend of thermogenic food ingredients containing African mango, citrus fruit extract, Coleus forskohlii, dihydrocapsiate, and red pepper was tested alone and in combination with a whey protein supplement for its effects on body composition in sedentary mice during high-fat diet. OBJECTIVE: The objective of this study was to evaluate the interaction of thermogenic foods on improving body composition during consumption of an unhealthy diet. MATERIALS AND METHODS: C57BL/6J young adult male mice (n = 12) were placed on a 60% high-fat diet for 4 weeks and subsequently randomly assigned to receive daily dosing by oral gavage of vehicle, the novel blend alone or with whey protein supplement for another 4 weeks. Body composition, thermal imaging of brown adipose tissue (BAT), mitochondrial BAT uncoupling protein 1 (UCP1), and plasma levels of leptin were assessed. RESULTS: Novel blend alone and in combination with protein supplement attenuated body weight gain, fat, and increased surface BAT temperature in comparison to vehicle control and to baseline (P < 0.5). The combination of novel blend and whey protein supplement also significantly increased UCP1 protein expression in BAT mitochondria in comparison to vehicle control and novel blend alone (P < 0.5). CONCLUSIONS: These data indicate that this novel blend stimulates thermogenesis and attenuates the gain in body weight and fat in response to high-fat diet in mice and these effects were improved when administered in combination with whey protein supplement. SUMMARY: 30 days oral administration to mice of a novel blend containing African mango seed extract, citrus fruits extract, Coleus forskohlii root extract, dihydrocapsiate and red pepper fruit extract reduced body weight and fat gain in response to high-fat diet without impairing muscle mass.The novel blend stimulated thermogenesis as shown by the increased thermal imaging and UCP1 protein expression in brown adipose tissue, indicating that improvement in body composition potentially occurred due to a fat-burning effect.The positive effects on body weight, fat, and thermogenesis were improved when the novel blend was administered in combination with a whey protein supplement suggesting that protein provides a synergistic fat-burning effect. Abbreviations Used: BAT: Brown adipose tissue, UCP1: Uncoupling protein 1, DEXA: Dual-energy X-ray absorptiometry.

Comparison of antimicrobial activities of natural essential oils and synthetic fragrances against selected environmental pathogens.

Plant essential oils (EOs) are known to inhibit the growth of bacteria and fungi. Whether these antimicrobial effects are comparable to synthetic household products is less clear. Furthermore, limited research is available on the potential additive effect of blending EOs. In this investigation, a new EO blend containing orange, patchouli, peppermint, and clary sage was compared to its individual single oils and to three household products-air freshener, liquid soap, and body spray-for their ability to inhibit the growth of Staphylococcus aureus, Streptococcus pneumoniae, Pseudonomas aeruginosa, and Aspergillus brasiliensis in the disc-diffusion assay. The new EO blend significantly inhibited the growth of the four microorganisms. The zones of inhibition of new EO blend were greater than the air freshener and similar to the liquid soap and body spray, with the exception of Str. pneumoniae in which the body spray provided greater inhibitory zone. The new EO blend and the single oils, with the exception of peppermint, equally inhibited the growth of S. aureus and Str. pneumoniae suggesting no additive effect. P. aeruginosa and A. brasiliensis showed variable susceptibility to all EOs except for no susceptibility to orange and limonene. No difference was found between (-) and (+)-limonene; whereas, (+)-menthol showed greater effect than (-)-menthol. In conclusion, blending the EO of orange, patchouli, peppermint, and clary sage was beneficial in inhibiting the growth of S. aureus, Str. pneumoniae, P. aeruginosa, and A. brasiliensis providing a natural antimicrobial fragrance option over synthetics fragrances used in soaps, body sprays, and air fresheners.

A small-molecule inhibitor of PIM kinases as a potential treatment for urothelial carcinomas.

The proto-oncogene proviral integration site for moloney murine leukemia virus (PIM) kinases (PIM-1, PIM-2, and PIM-3) are serine/threonine kinases that are involved in a number of signaling pathways important to cancer cells. PIM kinases act in downstream effector functions as inhibitors of apoptosis and as positive regulators of G1-S phase progression through the cell cycle. PIM kinases are upregulated in multiple cancer indications, including lymphoma, leukemia, multiple myeloma, and prostate, gastric, and head and neck cancers. Overexpression of one or more PIM family members in patient tumors frequently correlates with poor prognosis. The aim of this investigation was to evaluate PIM expression in low- and high-grade urothelial carcinoma and to assess the role PIM function in disease progression and their potential to serve as molecular targets for therapy. One hundred thirty-seven cases of urothelial carcinoma were included in this study of surgical biopsy and resection specimens. High levels of expression of all three PIM family members were observed in both noninvasive and invasive urothelial carcinomas. The second-generation PIM inhibitor, TP-3654, displays submicromolar activity in pharmacodynamic biomarker modulation, cell proliferation studies, and colony formation assays using the UM-UC-3 bladder cancer cell line. TP-3654 displays favorable human ether-à-go-go-related gene and cytochrome P450 inhibition profiles compared with the first-generation PIM inhibitor, SGI-1776, and exhibits oral bioavailability. In vivo xenograft studies using a bladder cancer cell line show that PIM kinase inhibition can reduce tumor growth, suggesting that PIM kinase inhibitors may be active in human urothelial carcinomas.

Pharmacologic activation of PKM2 slows lung tumor xenograft growth.

Inactivation of the M2 form of pyruvate kinase (PKM2) in cancer cells is associated with increased tumorigenicity. To test the hypothesis that tumor growth may be inhibited through the PKM2 pathway, we generated a series of small-molecule PKM2 activators. The compounds exhibited low nanomolar activity in both biochemical and cell-based PKM2 activity assays. These compounds did not affect the growth of cancer cell lines under normal conditions in vitro, but strongly inhibited the proliferation of multiple lung cancer cell lines when serine was absent from the cell culture media. In addition, PKM2 activators inhibited the growth of an aggressive lung adenocarcinoma xenograft. These findings show that PKM2 activation by small molecules influences the growth of cancer cells in vitro and in vivo, and suggest that such compounds may augment cancer therapies.

Discovery of Novel Putative Inhibitors of UDP-GlcNAc 2-Epimerase as Potent Antibacterial Agents.

We present the discovery and optimization of a novel series of inhibitors of bacterial UDP-N-acetylglucosamine 2-epimerase (called 2-epimerase in this paper). Starting from virtual screening hits, the activity of various inhibitory molecules was optimized using a combination of structure-based and rational design approaches. We successfully designed and identified a 2-epimerase inhibitor (compound 12-ES-Na, that we named Epimerox) which blocked the growth of methicillin-resistant Staphylococcus aureus (MRSA) at 3.9 µM MIC (minimum inhibitory concentration) and showed potent broad-range activity against all Gram-positive bacteria that were tested. Additionally a microplate coupled assay was performed to further confirm that the 2-epimerase inhibition of Epimerox was through a target-specific mechanism. Furthermore, Epimerox demonstrated in vivo efficacy and had a pharmacokinetic profile that is consonant with it being developed into a promising new antibiotic agent for treatment of infections caused by Gram-positive bacteria.

S110, a 5-Aza-2'-deoxycytidine-containing dinucleotide, is an effective DNA methylation inhibitor in vivo and can reduce tumor growth.

Methylation of CpG islands in promoter regions is often associated with gene silencing and aberrant DNA methylation occurs in most cancers, leading to the silencing of some tumor suppressor genes. Reversal of this abnormal hypermethylation by DNA methylation inhibitors is effective in reactivating methylation-silenced tumor suppressor genes both in vitro and in vivo. Several DNA methylation inhibitors have been well studied; the most potent among them is 5-aza-2'-deoxycytidine (5-Aza-CdR), which can induce myelosuppression in patients. S110 is a dinucleotide consisting of 5-Aza-CdR followed by a deoxyguanosine, which we previously showed to be effective in vitro as a DNA methylation inhibitor while being less prone to deamination by cytidine deaminase, making it a promising alternative to 5-Aza-CdR. Here, we show that S110 is better tolerated than 5-Aza-CdR in mice and is as effective in vivo in inducing p16 expression, reducing DNA methylation at the p16 promoter region, and retarding tumor growth in human xenograft. We also show that S110 is effective by both i.p. and s.c. deliveries. S110 therefore is a promising new agent that acts similarly to 5-Aza-CdR and has better stability and less toxicity.

In vivo transscleral iontophoresis of amikacin to rabbit eyes.

The objectives of these studies were to determine the amount and distribution of the aminoglycoside antibiotic amikacin delivered to rabbit eyes following transscleral iontophoresis and to determine the inter-study reproducibility of delivery over three identical studies. New Zealand White rabbits (N = 6 per dose group) were treated with a 200-mg/mL amikacin solution at 0, 2, 3 or 4 mA of (+) DC current for 20 minutes. Amikacin concentrations in eye tissues were highest with the 4-mA treatment. Concentrations for all three studies at this current were approximately 5.4, 40, 41, 343, and 92 mcg/g in the vitreous humor, anterior segment, non-treated hemisphere of the sclera, treated hemisphere of the sclera, and retina/choroid, respectively. These values were approximately 27, 50, 40, 10, and 13 fold greater than in the 0-mA control group and are well above the in vitro minimum inhibitory concentrations (MICs) for this drug. Inter-study reproducibility (measured as %CV) depended on the tissue type and treatment group and ranged from 8% for the retina/choroid to 51% for the anterior segment in the 4-mA group. Pretreatment with topical proparacaine hydrochloride local anesthetic did not affect amikacin delivery and total drug delivered was not affected by delivery time for the same total charge administered. Therapeutically relevant amounts of amikacin were delivered into eye tissues in a reproducible and controllable manner.