Six Weeks of Supplementation with Bovine Colostrum Effectively Reduces URTIs Symptoms Frequency and Gravity for Up to 20 Weeks in Pre-School Children.

Bovine colostrum is considered to provide anti-infective protection. Here, we present the first randomized controlled trial (RCT) aimed at assessing the preventive use of colostrum against upper respiratory tract infections (URTIs) in healthy pre-school children. We analyzed 57 children-35 in the colostrum (COL-dried bovine colostrum) and 22 in the placebo (PBO-dried whey) group, who received these substances as follows: first 15 days 2 × 500 mg and then 30 days 1 × 500 mg. The reporting on the children's health status, specifically on the frequency and gravity of URTI symptoms and abdominal side effects, was performed via an online survey. The influence of colostrum on the frequency of days with URTI symptoms remained significant until the 20th week of observation and reached 31% of median reduction. The median reduction reached 37% when the gravity of symptoms was analyzed. When we grouped symptomatic days into episodes of second gravity level, the reduction in their frequency was even larger (50%) and lasted until the end of the trial (21 weeks). No significant side effects, especially abdominal, were reported during the trial. Colostrum supplementation in pre-school children is well tolerated, safe and provides protection from frequency of URTIs and their gravity.

Moderate Dose Bovine Colostrum Supplementation in Prevention of Upper Respiratory Tract Infections in Medical University Students: A Randomized, Triple Blind, Placebo-Controlled Trial.

Colostrum supplementation has been confirmed to protect from upper respiratory tract infections (URTIs) in athletes. Our trial was designed to find out whether other young adults who have potentially been exposed to increased risk of developing URTIs can also benefit. Homogenous population of medical (MED) students (at risk) and health science (HSci) peers were supplemented with a relatively low dose (0.5-1.0 g/day) of bovine colostrum (COL) or placebo (PBO) over 45 days and then once again over 7 days starting at day 87. The trial lasted 107 days. Subjects were monitored solely by them filling out online daily questionnaires containing questions about frequency and severity of URTIs symptoms, well-being, and potential gastrointestinal side-effects. A significant level of protection from URTIs was observed as expressed by dropping frequency of symptomatic days in COL vs. PBO group among MED vs. HSci students. The same effect was also recorded for severity of symptoms, as well as general well-being perception. Overall, it can be concluded that although young healthy people seem to have sufficient defenses from URTIs, COL supplementation can provide significant support in such protection among those at higher infectious risk because of exposure to a heavy workload and increased contact with infectious agents.

Response of Skin-Derived and Metastatic Human Malignant Melanoma Cell Lines to Thymoquinone and Thymoquinone-Loaded Liposomes.

Thymoquinone has been proved to be effective against neoplasms, including skin cancer. Its high lipophilicity, however, may limit its potential use as a drug. Melanoma remains the deadliest of all skin cancers worldwide, due to its high heterogeneity, depending on the stage of the disease. Our goal was to compare the anti-cancer activity of free thymoquinone and thymoquinone-loaded liposomes on two melanoma cell lines that originated from different stages of this cancer: skin-derived A375 and metastatic WM9. We evaluated the proapoptotic effects of free thymoquinone by flow cytometry and Western blot, and its mitotoxicity by means of JC-1 assay. Additionally, we compared the cytotoxicity of free thymoquinone and thymoquinone in liposomes by WST-1 assay. Our results revealed a higher antiproliferative effect of TQ in WM9 cells, whereas its higher proapoptotic activity was observed in the A375 cell line. Moreover, the thymoquinone-loaded liposome was proved to exert stronger cytotoxic effect on both cell lines studied than free thymoquinone. Differences in the response of melanoma cells derived from different stages of the disease to thymoquinone, as well as their different responses to free and carrier-delivered thymoquinone, are essential for the development of new anti-melanoma therapies. However, further research is required to fully understand them.

Identified in blood diet-related methylation changes stratify liver biopsies of NAFLD patients according to fibrosis grade.

BACKGROUND: High caloric diet and lack of physical activity are considered main causes of NAFLD, and a change in the diet is still the only effective treatment of this disease. However, molecular mechanism of the effectiveness of diet change in treatment of NAFLD is poorly understood. We aimed to assess the involvement of epigenetic mechanisms of gene expression regulation in treatment of NAFLD. Eighteen participants with medium- to high-grade steatosis were recruited and trained to follow the Mediterranean diet modified to include fibre supplements. At three timepoints (baseline, after 30 and 60 days), we evaluated adherence to the diet and measured a number of physiological parameters such as anthropometry, blood and stool biochemistry, liver steatosis and stiffness. We also collected whole blood samples for genome-wide methylation profiling and histone acetylation assessment. RESULTS: The diet change resulted in a decrease in liver steatosis along with statistically significant, but a minor change in BMI and weight of our study participants. The epigenetic profiling of blood cells identified significant genome-wide changes of methylation and acetylation with the former not involving regions directly regulating gene expression. Most importantly, we were able to show that identified blood methylation changes occur also in liver cells of NAFLD patients and the machine learning-based classifier that we build on those methylation changes was able to predict the stage of liver fibrosis with ROC AUC = 0.9834. CONCLUSION: Methylomes of blood cells from NAFLD patients display a number of changes that are most likely a consequence of unhealthy diet, and the diet change appears to reverse those epigenetic changes. Moreover, the methylation status at CpG sites undergoing diet-related methylation change in blood cells stratifies liver biopsies from NAFLD patients according to fibrosis grade.

Post-Delivery Milking Delay Influence on the Effect of Oral Supplementation with Bovine Colostrum as Measured with Intestinal Permeability Test.

Background and objective: The health supplement bovine colostrum reportedly improves immunity and regulates intestinal homeostasis. Reliable assessment methods are needed to ensure the satisfactory biological activity of all marketed colostrum products. Of the well-established effects of colostrum use, the restoration of appropriate intestinal permeability assessed with the lactulose/mannitol (L/M) differential sugar absorption test upon supplementation with colostrum has been consistently observed. Milking time after delivery is one of the factors that influences the composition of bovine colostrum, which causes a rapid decrease in bioactive components. Materials and methods: We use the L/M test to evaluate the intestinal permeability reduction upon supplementation with colostrum (2 × 500 mg) harvested at various times after delivery (2, 24, and 72 h) or a placebo (whey). In our randomized, double-blind placebo-controlled (DBPC) trial, 31 healthy athletes were divided into four groups and assessed at baseline and after the intervention. Results: The trial revealed that only colostrum collected after 2 h and 24 h caused a significant reduction of intestinal permeability. The comparison of post-intervention vs. baseline Δ values produced statistically significant results for 2 h colostrum versus the placebo and 72 h colostrum groups. Conclusions: We conclude that the change of bovine colostrum composition over the first three days of lactation is accompanied by a decrease in its biological activity as measured with the L/M test. This test may offer a biological quality measure for colostrum.

Assessing the Association of Elevated Zonulin Concentration in Stool with Increased Intestinal Permeability in Active Professional Athletes.

BACKGROUND AND OBJECTIVES: The causative factors or conditions leading to increased intestinal permeability (IIP) have only been partly elucidated, suggesting excessive zonulin release to be a key factor among them. Likewise, it is known that athletic activity predisposes individuals towards the development of IIP; however, little is understood about the nature of this phenomenon. We decided to test the actual coincidence between IIP and increased stool zonulin (ISZ) in actively training athletes. Materials and Methods: We compared intestinal permeability tested with lactulose/mannitol differential absorption (lactulose/mannitol (L/M) test) and zonulin concentration in stool in 20 professional athletes (PRO), 9 amateur athletes (AMA), and 9 non-athletes (CTR). Results: The results confirmed that professional athletic activity showed significant positive association with intestinal permeability. ISZ was observed exclusively in athletes (CTR vs. AMA vs. PRO, respectively, 0% vs. 22% vs. 55%), and its prevalence was significantly higher in PRO than CTR. When we divided the participants into four categories related to exceeding the upper reference limits for both tested parameters (ISZ + or - and IIP + or -), significant differences were found between CTR and PRO; however, no significant differences were found between CTR and AMA or AMA and PRO. Conclusions: Our trial confirmed previous findings that professional athletic activity predisposes individuals to IIP. We also demonstrated that although ISZ was associated with intense training, there was no statistically significant association between ISZ and IIP in the tested group of professional athletes, which suggests the existence of additional mechanisms causing IIP.

Oral Supplementation with Bovine Colostrum Decreases Intestinal Permeability and Stool Concentrations of Zonulin in Athletes.

Increased intestinal permeability has been implicated in various pathologies, has various causes, and can develop during vigorous athletic training. Colostrum bovinum is a natural supplement with a wide range of supposed positive health effects, including reduction of intestine permeability. We assessed influence of colostrum supplementation on intestinal permeability related parameters in a group of 16 athletes during peak training for competition. This double-blind placebo-controlled study compared supplementation for 20 days with 500 mg of colostrum bovinum or placebo (whey). Gut permeability status was assayed by differential absorption of lactulose and mannitol (L/M test) and stool zonulin concentration. Baseline L/M tests found that six of the participants (75%) in the colostrum group had increased intestinal permeability. After supplementation, the test values were within the normal range and were significantly lower than at baseline. The colostrum group Δ values produced by comparing the post-intervention and baseline results were also significantly lower than the placebo group Δ values. The differences in stool zonulin concentration were smaller than those in the L/M test, but were significant when the Δ values due to intervention were compared between the colostrum group and the placebo group. Colostrum bovinum supplementation was safe and effective in decreasing of intestinal permeability in this series of athletes at increased risk of its elevation.

Mixed chimerism and transplant tolerance are not effectively induced in C3a-deficient mice.

Mixed chimerism, a phenomenon involved in the development of specific alloantigen tolerance, could be achieved through the transplantation of hematopoietic stem cells into properly prepared recipients. Because the C3a complement component modulates hematopoietic cell trafficking after transplantation, in the present study, we investigated the influence of the C3a deficiency on mixed chimerism and alloantigen tolerance induction. To induce mixed chimerism, C57BL/6J (wild-type strain; H-2K(b); I-E(-)) and B6.129S4-C3(tm1Crr)/J (C3a-deficient) mice were exposed to 3 G total body irradiation (day -1). Subsequently, these mice were treated with CD8-blocking (day -2) and CD40L-blocking (days 0 and 4) antibodies, followed by transplantation with 20 × 10(6) Balb/c (H-2K(d); I-E(+)) bone marrow cells (day 0). The degree of mixed chimerism in peripheral blood leukocytes was measured several times during the 20-week experiment. The tolerance to Balb/c mouse antigens was assessed based on the number of lymphocytes expressing Vß5 and Vß11 T-cell receptor and on skin-graft (day 0) acceptance. Applying our experimental model, mixed chimerism and alloantigen tolerance were effectively induced in C57BL/6J (wild-type) mice, but not in C3a(-/-) animals. The present study is, to our knowledge, the first to demonstrate that C3a is vital for achieving stable mixed chimerism and related to this induction of transplant tolerance.

Different strategies of mixed chimerism induction may determine stem/progenitor cell populations in recipient mice.

Mixed chimerism has been suggested to induce tolerance to transplanted alloantigens. As the precise influence of mixed chimerism induction on the host organism has still not been fully elucidated, the aim of the present study was to explore this phenomenon in relation to the stem cell compartment. The experiment was performed on B6.SJL-Ptprc(a)Pep3(b) mice. Mixed chimerism induction protocols involved 3 Gy TBI (Day -1 of the experiment), injection of 20-30 × 10(6) Balb C bone marrow cells (Day 0), and administration of blocking antibodies against CD40L (Day 0 and Day 4), anti-CD8 (Day -2) with/without anti-NK1.1 (Day -3). Selected groups of mice were also treated with cyclophosphamid (175 mg/kg) on Day 2. The presence of mixed chimerism was assessed in peripheral blood, bone marrow, and spleen, as well as in various subpopulations of leukocytes (CD4(+), CD8(+), CD45/B220(+), Gr-1(+), lin(-)/Sca-1(+)/c-kit(-), lin(-)/Sca-1(+)/c-kit(+), lin(-)/Sca-1(-)/c-kit(+)). Furthermore, the percentage of stem/progenitor cells (lin(-)/Sca-1(+)/c-kit(-), lin(-)/Sca-1(+)/c-kit(+), lin(-)/Sca-1(-)/c-kit(+), VSEL, HSC) was analysed for the first time in bone marrow and peripheral blood of chimeric mice. The range of mixed chimerism differed significantly among various cell populations: it was lowest in CD8-positive cells and lin(-)/Sca-1(+)/c-kit(-) cells, and highest in granulocytes. The induction of mixed chimerism revealed a significant impact on the stem/progenitor cell frequency in recipient mice, providing potential therapeutic insights into the long-term immunologic tolerance observed in chimeric mice. Collectively, these findings contribute to further optimization of mixed chimerism induction protocols and might help in the introduction of this phenomenon into clinical practice.