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Background Bovine lactoferrin is increasingly being used as an ingredient in infant formula manufacture to enhance nutritional efficacy through the provision of growth, immunoprotective, and antimicrobial factors to the neonate. Objective To evaluate method reproducibility of AOAC First Action Official Method   2021.07 for compliance with the performance requirements described in Standard Method Performance Requirement (SMPR®) 2020.005. Methods Eight laboratories participated in the analysis of blind-duplicate samples of seven nutritional products. Samples were diluted in buffer, and an optical biosensor immunoassay was used in a direct-assay format to quantitate bovine lactoferrin by its interaction with an immobilized anti-lactoferrin antibody. Quantitation was accomplished by the external standard technique with interpolation from a four-parameter calibration regression. Results After outliers were removed, precision as reproducibility was found to be within limits set in SMPR 2020.005 (≤ 9%) for six out of seven samples and all had acceptable Horwitz Ratio (HorRatR) values ranging from 1.0 to 2.1. Additionally, comparison with an alternative independent Stakeholder Panel on Infant Formula and Adult Nutritionals (SPIFAN) First Action method (heparin cleanup LC–UV), showed negligible difference between results. Conclusion The method described is suitable for the quantification of intact, undenatured bovine lactoferrin in powdered infant formulas. The SPIFAN Expert Review Panel evaluated the method and accompanying validation data from this multi-laboratory testing (MLT) study in July 2023 and recommended Official Method   2021.07 for adoption as a Final Action Official Method SM. Highlights A multi-laboratory validation study of an automated optical biosensor immunoassay for the determination of intact, undenatured bovine lactoferrin is described. [ABSTRACT FROM AUTHOR]

Lactoferrin is an antimicrobial glycoprotein that demonstrates a broad-spectrum of activity against a wide variety of clinical pathogens. This study investigated the potential of bovine lactoferrin (bLf) against multidrug resistant Staphylococcus capitis (S. capitis) strains. Growth curve analysis and time-kill curves demonstrated that at 750 µg ml−1 lactoferrin significantly inhibited (50.6%, P < 0.05) the growth of most isolates tested (90%), and this effect was based on a bacteriostatic mechanism. At the same concentration, bLf also significantly inhibited (30%, P < 0.05) biofilm formation in 40% of strains tested. Combinations of bLf with selected antibiotics were assessed for enhanced antimicrobial activity using growth curves. BLf combined with β-lactam antibiotics reduced the growth of S. capitis strains, however, the effects were not significant. BLf displays antimicrobial effects against multidrug resistant S. capitis isolates, but with strain-specific effects. [ABSTRACT FROM AUTHOR]

Background Infant formulas, and pediatric and adult nutritional products, are being fortified with bovine lactoferrin (bLF) due to its beneficial impacts on immune development and gut health. Lactoferrin supplementation into these products requires an analytical method to accurately quantify the concentrations of bLF to meet global regulatory and quality standards. Objective To develop and validate a lactoferrin method capable of meeting the AOAC INTERNATIONAL Standard Method Performance Requirements (SMPR®) 2020.005. Methods Powder formula samples are extracted using warm dibasic phosphate buffer, pH 8, then centrifuged at 4°C to remove insoluble proteins, fat, and other solids. The soluble fraction is further purified on a HiTrap heparin solid-phase extraction (SPE) column to isolate bLF from interferences. Samples are filtered, then analyzed by LC–UV using a protein BEH C4 analytical column and quantitated using an external calibrant. Results The LOQ (2 mg/100 g), repeatability (RSD: 2.0–4.8%), recovery (92.1–97.7%), and analytical range (4–193 mg/100 g) all meet the method requirements as stated in SMPR 2020.005 for lactoferrin. Conclusion The reported single-laboratory validation (SLV) results demonstrate the ability of this lactoferrin method to meet or exceed the method performance requirements to measure soluble, intact, non-denatured bLF in infant and adult nutritional powder formulas. Highlights The use of a heparin affinity column to isolate lactoferrin from bovine milk products combined with a selective analytical chromatographic column provides suitable analyte specificity without requiring proprietary equipment or reagents. [ABSTRACT FROM AUTHOR]

A key molecule for neutrophil degranulation is Rac2 guanosine triphosphatase. Neutrophils from Rac2 knockout mice (Rac2−/−) exhibit impaired primary granule exocytosis in response to cytochalasin B/f-Met-Leu-Phe, while secondary and tertiary granule release is unaffected. Coronin 1A, a protein involved in actin remodeling, is diminished in Rac2−/− neutrophils. However, primary granule exocytosis from Rac2−/− neutrophils has not been determined using more immunologically relevant stimuli. We sought to determine the role of Rac2 in degranulation and actin cytoskeleton rearrangement in response to immobilized immune complexes and relate this to intracellular coronin 1A localization. We used bone marrow neutrophils from wild-type and Rac2−/− mice stimulated with immobilized immune complexes. Secretion of primary (myeloperoxidase), secondary (lactoferrin), and tertiary granule (MMP-2 and MMP-9) products was evaluated. Subcellular colocalization of coronin 1A with actin and the primary granule marker CD63 was determined by deconvolution microscopy. We found major differences in myeloperoxidase, MMP-2, and MMP-9 but not lactoferrin release, along with diminished filopodia formation, CD63 polarization, and colocalization of coronin 1A with CD63 in immune complex–stimulated Rac2−/− bone marrow neutrophils. Rac2 and coronin 1A were found associated with granules in cytochalasin B/f-Met-Leu-Phe–activated human neutrophils. This report confirms a role for Rac2 in immunologically relevant stimulation of neutrophil granule exocytosis. Rac2 appears to attach to neutrophil granules, polarize CD63+ granules to the cell surface in a manner dependent on coronin 1A, and induce filopodia formation. Our studies provide insight into mechanisms of Rac2-mediated regulation of granule exocytosis. Activation of Rac2 guanosine triphosphatase is essential for exocytosis of primary and tertiary granules in response to immune complexes. [ABSTRACT FROM AUTHOR]

Keywords: colostrum; FcRn; lactoferrin; mammary; regulation EN colostrum FcRn lactoferrin mammary regulation 37 43 7 06/20/23 20230601 NES 230601 Implications Colostrum formation is directed by placental structure and hormone signals Quality colostrum is defined by the concentration of immunoglobulin Quality colostrum formation in ungulates (hoofed mammals) is dependent on the Fc receptor of the neonate (FcRn) While the FcRn binds IgG1, IgG2, and albumin, only IgG1 is transported to colostrum FcRn IgG release into colostrum is pH dependent and lactoferrin likely facilitates it release Introduction Members of the class mammalia are so named for their mammary glands. Another example of this variation is that ungulate species form colostrum with IgG in a period of 3-4 weeks before birth ending shortly after partum, while the rodent species continue colostrum production over the entire suckling period (21 d) with an IgA emphasis that gradually transitions to mature milk. Immunoglobulins present in cow colostrum include G, M, and A, but it is IgG subclass 1 (IgG1) that constitutes 65-80% of immunoglobulin in cow colostrum (Table 1). Thus, milk secretion and hence dilution of the pre-colostrum secretion starts at this time and provides liquid colostrum at first milking ([14]). [Extracted from the article]

Equine milk has 21% less calcium and 20% less phosphorus than bovine milk, but equine milk contains 66% more calcium and 60% more phosphorus than human milk. The essential amino acid composition in equine milk is similar to that of bovine and human milk; however, while equine milk contains more essential amino acids compared with cows' milk, human milk contains 10× greater amounts of essential amino acids than equine. Influence of stage of lactation and year season on composition of mares' colostrum and milk and method and time of storage on vitamin C content in mares' milk. Keywords: colostrum; horse; lactation; milk EN colostrum horse lactation milk 96 100 5 06/20/23 20230601 NES 230601 Implications Mare colostrum contains immunoglobulin G and bioactive factors critical to the health of the newborn foal. [Extracted from the article]

Background: Anemia is one of the main problems in hemodialysis patients, which is caused by inadequate production of erythropoietin. Functional iron deficiency (FID) anemia is a kind of anemia, which lack of functional iron therein leads to resistance and inappropriate response to erythropoietin in hemodialysis patients. Aim of the Work: The aim of the present study was to study the effect and safety of oral lactoferrin therapy for functional iron deficiency anemia in prevalent hemodialysis patients. Patients and Methods: This was Randomized clinical trial, was conducted at El Demerdash hemodialysis unit on 70 prevalent hemodialysis patients divided into 2 groups: (Group A): included 35 patients with functional iron deficiency anemia, received lactoferrin together with the recommended dose of ESA therapy for 3 months, (Group B): included 35 patients with functional iron deficiency anemia, received no lactoferrin apart from the recommended dose of ESA therapy for 3 months. Results: There was no statistically significant difference between the two studied groups (group A and group B) as regard sex, occupation, residence and age. There was no statistically significant difference between the two studied groups (group A and group B) as regard height, weight and BMI. There was no statistically significant difference between the two studied groups (group A and group B) as regard history. There was no statistically significant difference between the two studied groups (group A and group B) as regard hemoglobin and WBCs. There was no statistically significant difference between the two studied groups (group A and group B) as regard PLT and Reticulocyte %. There was no statistically significant difference between the two studied groups (group A and group B) as regard ferritin, serum iron and TSAT. There was highly statistically significant difference between the two studied groups (group A and group B) as regard KT/V and URR at 3 months. There was nostatistically significant difference between the two studied groups (group A and group B) as regard KT/V and URR at baseline. Conclusion: Oral lactoferrin could not be used as a substitute to totally replace IV ferrous sulphate in treating iron deficiency anemia in end stage renal disease (ESRD) on dialysis. Oral lacto-ferrin could not even control Hb level constant without dropping in patients with end stage renal disease (ESRD). Oral lactoferrin had some gastrointestinal effects on some patients which did not allow compliance on the medication. [ABSTRACT FROM AUTHOR]

Introduction: acute lung injury (ALI) is one of the most critical illnesses affecting the lung and is commonly associated with severe complications. management of ALI without major side effects remains a continuous challenge. Lactoferrin (LF) is an iron-binding glycoprotein having potential anti-inflammatory effects. Aim: the study was conducted to compare the therapeutic role of lactoferrin (LF) versus dexamethasone (DEXA) on the microscopic structure of the lung in a model of lipopolysaccharide (LP) induced acute lung injury. Material and Methods: thirty-five adult male albino rats weighing (180-200gm) were divided randomly into 4 groups. Group I (control group), Group II (LP group): rats received a single IP injection of Lipopolysaccharide (3mg/kg). Group III (LP+LF group): rats received LP as in group II followed one hour later by IP injection of lactoferrin (5mg/rat). Group IV (LP+DEXA group): rats received LP as in group II followed one hour later by IP injection of dexamethasone (5mg/kg). after 24 hours, lung specimens were proceeded for light microscopic examination, as well as Morphometrical and statistical analyses. Results: H&E-stained section of Group II showed marked inflammatory changes, interalveolar septa were significantly thickened and were heavily infiltrated with inflammatory cells and extravasated red blood cells were occasionally detected. Many alveoli appeared collapsed. The lining epithelium of bronchi appeared occasionally desquamated and peri-bronchial lymphoid aggregation were observed. An apparent increase in mucus production and significant increase in number of goblet cells were detected by PAS-stain. Orcein-stained section showed disruption of elastic fibers. immunohistochemical stain revealed a significantly increased reaction to iNOS antibodies. On the other hand, the lung structure in group III and group IV were much more improved, with group IV very similar to the control group. Conclusion: Both lactoferrin and dexamethasone showed therapeutic and anti-inflammatory effects in LP induced ALI. These effects were more prominent in dexamethasone than lactoferrin as detected by microscopic, morphometric, and statistical studies. [ABSTRACT FROM AUTHOR]

Background: Bovine lactoferrin is increasingly being used as an ingredient in infant formula manufacture to enhance nutritional efficacy through the provision of immunoprotective, growth, and antimicrobial factors to the neonate. Objective: To evaluate the analytical performance of an optical biosensor immunoassay for compliance with the method performance requirements described in SMPR 2020.005. Method: Following dilution of the sample in buffer, an automated, label-free, real-time optical biosensor immunoassay was used in a direct assay format to quantitate bovine lactoferrin by its interaction with an immobilized anti-lactoferrin antibody. Quantitation was accomplished by the external standard technique with interpolation from a 4-parameter calibration regression. Results: The analytical range (0-200 mg/hg), method detection limit (0.8 mg/hg), recovery (96.1-109.2%), and repeatability (1.0-5.3%) complied with the requirements given in the lactoferrin SMPR. The method was shown to be specific for native, intact lactoferrin; thermally denatured lactoferrin generated no measurable binding response. Conclusion: The method described is suitable for the quantification of intact, undenatured lactoferrin in milk products, infant formulas (bovine milk protein-based, soy protein-based, and amino acid-based), and adult nutritionals and has been demonstrated to meet the performance requirements defined in SMPR 2020.005. Highlights: A single-laboratory validation (SLV) of an automated biosensor immunoassay for the determination of intact, undenatured lactoferrin is described. [ABSTRACT FROM AUTHOR]

Transitory appearance of immune suppressive polymorphonuclear neutrophils (PMNs) defined as myeloid‐derived suppressor cells (PMNs‐MDSCs) in newborns is important for their protection from inflammation associated with newly established gut microbiota. Here, we report that inhibition of the type I IFN (IFN1) pathway played a major role in regulation of PMNs‐MDSCs‐suppressive activity during first weeks of life. Expression of the IFN1 receptor IFNAR1 was markedly lower in PMNs‐MDSCs. However, in newborn mice, down‐regulation of IFNAR1 was not sufficient to render PMNs immune suppressive. That also required the presence of a positive signal from lactoferrin via its receptor low‐density lipoprotein receptor‐related protein 2. The latter effect was mediated via NF‐κB activation, which was tempered by IFN1 in a manner that involved suppressor of cytokine signaling 3. Thus, we discovered a mechanism of tight regulation of immune suppressive PMNs‐MDSCs in newborns, which may be used in the development of therapies of neonatal pathologies. [ABSTRACT FROM AUTHOR]

Tooth decay is one of the most common diseases that humans face during their lifetime. Streptococcus mutans is one of the most critical factors of early tooth decay. These bacteria may produce biofilms in the mouth, which are more resistant to antimicrobial agents. Streptococcus mutans may also demineralize tooth enamel by producing lactic acid. In this study, the effect of nanoliposomes containing lactoferrin at three different concentrations (1·5, 3 and 6 mg ml−1) on the biofilm formed by S. mutans was investigated. Nanoliposomes were prepared using the thin‐layer hydration method. An active attachment model was used to evaluate biofilm and lactic acid production. The results showed that nano‐encapsulated lactoferrin could reduce CFU of biofilm more effectively than free lactoferrin. Lactoferrin‐containing nanoliposomes also significantly reduced lactic acid production by S. mutans. Therefore, nano‐encapsulated lactoferrin may be used along with other dental caries control methods to increase anti‐caries efficacy. [ABSTRACT FROM AUTHOR]

The significance of dairy in human health and nutrition is gaining significant momentum as consumers continue to desire wholesome, nutritious foods to fulfill their health and wellness needs. Bovine milk not only consists of all the essential nutrients required for growth and development, it also provides a broad range of bioactive components that play an important role in managing human homeostasis and immune function. In recent years, milk bioactives, including α-lactalbumin, lactoferrin, glycomacropeptide, milk fat globule membrane, and milk oligosaccharides, have been intensively studied because of their unique bioactivity and functionality. Challenges for the application of these bioactive components in food and pharmaceutical formulations are associated with their isolation and purification on an industrial scale and also with their physical and chemical instability during processing, storage, and digestion. These challenges can be overcome by advanced separation techniques and sophisticated nano- or micro-encapsulation technologies. Current knowledge about the chemistry, separation, and encapsulation technology of major bioactives derived from bovine milk and their application in the food industry is reviewed here. [ABSTRACT FROM AUTHOR]

Recent studies have demonstrated that the formation of an implantation chamber composed of a uterine crypt, an implantation-competent blastocyst, and uterine glands is a critical step in blastocyst implantation in mice. Leukemia inhibitory factor (LIF) activates signal transducer and activator of transcription 3 (STAT3) precursors via uterine LIF receptors (LIFRs), allowing successful blastocyst implantation. Our recent study revealed that the role of epithelial STAT3 is different from that of stromal STAT3. However, both are essential for blastocyst attachment, suggesting the different roles of epithelial and stromal LIFR in blastocyst implantation. However, how epithelial and stromal LIFR regulate the blastocyst implantation process remains unclear. To investigate the roles of LIFR in the uterine epithelium and stroma, we generated Lifr -floxed/lactoferrin (Ltf) -iCre (Lifr eKO) and Lifr- floxed/antimüllerian hormone receptor type 2 (Amhr2) -Cre (Lifr sKO) mice with deleted epithelial and stromal LIFR, respectively. Surprisingly, fertility and blastocyst implantation in the Lifr sKO mice were normal despite stromal STAT3 inactivation. In contrast, blastocyst attachment failed, and no implantation chambers were formed in the Lifr eKO mice with epithelial inactivation of STAT3. In addition, normal responsiveness to ovarian hormones was observed in the peri-implantation uteri of the Lifr eKO mice. These results indicate that the epithelial LIFR-STAT3 pathway initiates the formation of implantation chambers, leading to complete blastocyst attachment, and that stromal STAT3 regulates blastocyst attachment without stromal LIFR control. Thus, uterine epithelial LIFR is critical to implantation chamber formation and blastocyst attachment. [ABSTRACT FROM AUTHOR]

Background Human milk oligosaccharides (HMOs) and bioactive proteins likely benefit infant health, but information on these relations is sparse. Objectives We aimed to examine associations of milk content of HMOs and bioactive proteins with incidence and longitudinal prevalence of infant morbidity (any illness, fever, diarrhea, acute respiratory infection, and loss of appetite) and markers of inflammation [C-reactive protein (CRP) and α-1-acid glycoprotein (AGP)]. These are secondary analyses of a randomized controlled trial. Methods Breast milk samples at 6 mo postpartum (n   = 659) were analyzed to quantify absolute abundance of HMOs, relative abundance of fucosylated HMOs, sialylated HMOs, and 51 individual HMOs, and concentrations of 6 bioactive proteins (lactalbumin, lactoferrin, lysozyme, antitrypsin, IgA, and osteopontin). We examined associations of these constituents with infant morbidity from 6 to 7 and 6 to 12 mo, and CRP and AGP at 6 and 18 mo, considering maternal secretor status [presence or absence of the functional enzyme encoded by the fucosyltransferase 2 gene (FUT2) ] and adjusting for covariates and multiple hypothesis testing. Results In secretors there were positive associations between total HMOs and longitudinal prevalence of fever (P  = 0.032), between fucosylated HMOs and incidence of diarrhea (P  = 0.026), and between lactoferrin and elevated CRP at 18 mo (P  = 0.011). In nonsecretors, there were inverse associations between lactoferrin and incidence of fever (P   = 0.007), between osteopontin and longitudinal prevalence of lost appetite (P   = 0.038), and between fucosylated HMOs and incidence of diarrhea (P  = 0.025), lost appetite (P  = 0.019), and concentrations of AGP and CRP at 6 mo (P  = 0.001 and 0.010); and positive associations between total HMOs and incidence of lost appetite (P  = 0.024) and elevated CRP at 18 mo (P   = 0.026), between lactalbumin and incidence of diarrhea (P  = 0.006), and between lactoferrin and elevated CRP at 18 mo (P  = 0.015). Conclusions Certain HMOs and bioactive proteins were associated with infant morbidity and inflammation, particularly in nonsecretors. Further research is needed to elucidate the causality of these relations. This trial was registered at clinicaltrials.gov as NCT01239693. [ABSTRACT FROM AUTHOR]

Background Milk fat globule membrane (MFGM) and lactoferrin (LF) are human-milk bioactive components demonstrated to support gastrointestinal and immune development. Significantly fewer diarrhea and respiratory-associated adverse events through 18 mo of age were previously reported in healthy term infants fed a cow-milk–based infant formula with an added source of bovine MFGM and bovine LF through 12 mo of age. Objectives The aim was to compare microbiota and metabolite profiles in a subset of study participants. Methods Stool samples were collected at baseline (10–14 d of age) and day 120. Bacterial community profiling was performed via 16S rRNA gene sequencing and alpha and beta diversity were analyzed (QIIME 2). Differentially abundant taxa were determined using linear discriminant analysis effect size (LefSE) and visualized (Metacoder). Untargeted stool metabolites were analyzed (HPLC/MS) and expressed as the fold-change between group means (control to MFGM+LF ratio). Results Alpha diversity increased significantly in both groups from baseline to 4 mo. Subtle group differences in beta diversity were demonstrated at 4 mo (Jaccard distance; R 2 = 0.01, P  = 0.042). Specifically, Bacteroides uniformis and Bacteroides plebeius were more abundant in the MFGM+LF group at 4 mo. Metabolite profile differences for MFGM+LF versus control included lower fecal medium-chain fatty acids, deoxycarnitine, and glycochenodeoxycholate, and some higher fecal carbohydrates and steroids (P  < 0.05). After applying multiple test correction, the differences in stool metabolomics were not significant. Conclusions Addition of bovine MFGM and LF in infant formula was associated with subtle differences in stool microbiome and metabolome by 4 mo of age, including increased prevalence of Bacteroides species. Stool metabolite profiles may be consistent with altered microbial metabolism. This trial was registered at https://clinicaltrials.gov as NCT02274883. [ABSTRACT FROM AUTHOR]

The molecular mechanism of autophagy in Lactoferrin (LF) induced osteoblast differentiation is not fully demonstrated. In this study, alkaline phosphatase (ALP) activity, alizarin red S staining and ELISA were used to study N-terminal propeptide of type I procollagen (PINP) expression. mRFP-GFP-LC3 adenoviruses, mono-dansylcadaverine (MDC) staining, scanning electron microscopy, and western blot analysis was employed to probe the LF induced autophagy. The interaction between autophagy receptor Neighbor of Brca1 gene (Nbr1) and pp38 was studied. 3-methyladenine (3-MA) and chloroquine (CQ) could inhibit the activity of ALP, PINP and the autophagy in LF group. LF treatment could up-regulate and down-regulate the expressions of pp38 and Nbr1with a dose-dependent manner, respectively. LF could inhibit the recognition of pp38 and Nbr1. In addition, LF can prompt Nbr1-medicated autophagy and prevent pp38 degradation by autophagy. LF can induce Nbr1-mediated autophagy and inhibit pp38 entering into autophagy flux in the physiological process of osteoblast differentiation. Abbreviations: CQ:chloroquine；LF: Lactoferrin; 3-MA: 3-methyladenine; ALP: Alkaline phosphatase; ANOVA: Analysis of variance; CCK-8: Cell Counting Kit-8; LC3: Microtubule-associated protein light chain3; MDC: Monodansylcadaverine; Nbr1: neighbor of Brca1 gene; PINP: N-terminal propeptide of type I procollagen; PVDF: Polychlorotrifluoroethylene; pp38: phosphorylation p38; RAPA: Rapamycin; SDS: sodium dodecyl sulfate. LF can induce Nbr1-mediated autophagy and inhibit pp38 entering into autophagy flux in the physiological process of osteoblast differentiation. [ABSTRACT FROM AUTHOR]

The current study investigates anti-neoplastic and immunomodulatory activities of co-treatment based on bovine lactoferrin (bLF) and/or muramyl dipeptide (MDP) with or without cisplatin (Cis) in tumor-bearing mice. In the present study, bLF (100 mg/kg; orally) and MDP (0.5 mg/kg; subcutaneously) was administered alone or together.MDP or bLF was co-treated with Cis (1 mg/kg; intraperitoneally) in mice-bearing Ehrlich solid carcinoma. Tumor size, tumor mass proliferation, apoptosis using immunohistochemistry, the alteration in spleen cell proliferation, phenotype using f low cytometry and white blood cells total and differential counts were detected. Treatment with Cis or (bLF and MDP) significantly reduced tumor size, upregulated the pro-apoptotic p53 expression and downregulated the anti-apoptotic Bcl-2 and proliferative marker PCNA expression compared to non-treated tumor-bearing animals. Moreover, co-treatment of MDP and Cis significantly potentiated the reduction of the tumor size, downregulated the Bcl-2 and PCNA expression and upregulated the p53 expression compared to Cis-treated animals. While bLF and Cis co-treatment positively controlled PCNA and p53 expression compared to tumor-bearing animals, it significantly potentiated the reduction of the tumor size and downregulated the Bcl-2 expression compared to Cis-treated animals. Co-treatment of (bLF and MDP), (bLF and Cis) or (MDP and Cis) increased the spleen cell proliferation and altered the immunological profile of the CD3+CD4+, CD3+CD8+, CD3+CD4+CD69+, CD3+CD8+CD69+ and CD11b+Ly6G+ cells to achieve better immune response against tumor. In conclusion, co-treatments based on bLF and/or MDP are promising therapies against cancer, through their potency to control proliferation, enhance apoptosis and improve the immune status against tumor cells. [ABSTRACT FROM AUTHOR]

Lactoferrin (LF) is a sialylated iron-binding glycoprotein, occurring in several biological secretions like milk, saliva, and seminal fluids and is a major component of a mammalian innate immune system. It plays multiple protective roles against large group of microorganisms and performs anti-inflammatory and anti-cancer activities. The concentration of serum LF in gilt (primiparous sow) and their piglets remains unknown. We determined serum LF concentration in gilts during gestation and lactation to that of 19-d-old piglets, including sow-fed and formula-fed piglets using enzyme-linked immunosorbent assay (ELISA). We found that the concentration of serum LF in gilts varied during gestation (0.77 ± 0.10 µg/mL) and lactation (0.62 ± 0.11 µg/mL). The mean concentration of serum LF in gilts (0.72 ± 0.06 µg/mL) was significantly higher than that of piglets (0.42 ± 0.07 µg/mL, P = 0.004). Additionally, a marginal significant difference (P =0.06) was observed for serum LF concentration in sow-fed piglets (0.42 ± 0.03 µg/mL) at 19 d old compared to that of formula-fed piglets (0.33 ± 0.04 µg/mL) at 37 d old. This study provides noble information regarding the serum LF concentration in the healthy gilts and piglets and thereby the data can be used as a standard reference point for future studies on the role of LF in pig reproduction. [ABSTRACT FROM AUTHOR]

Lactoferrin modulates mucosal immunity and targets mechanisms contributing to inflammation during human immunodeficiency virus disease. A randomized placebo-controlled crossover clinical trial of recombinant human (rh) lactoferrin was conducted among 54 human immunodeficiency virus-infected participants with viral suppression. Outcomes were tolerability, inflammatory, and immunologic measures, and the intestinal microbiome. The median age was 51 years, and the median CD4+ cell count was 651/µL. Adherence and adverse events did not differ between rh-lactoferrin and placebo. There was no significant effect on plasma interleukin-6 or D-dimer levels, nor on monocyte/T-cell activation, mucosal integrity, or intestinal microbiota diversity. Oral administration of rh-lactoferrin was safe but did not reduce inflammation and immune activation. Clinical Trials Registration: NCT01830595. [ABSTRACT FROM AUTHOR]

Lactoferrin (LF) is a naturally occurring iron-binding glycoprotein with a variety of biological functions. It has increasing demand every year and huge market potential. In this study, we explored the feasibility of expressing human LF (hLF) in edible algae C. reinhardtii. A codon-optimized hLF gene was synthesized, inserted into pCAMBIA-1301C and transformed into C. reinhardtii SP strain. In total, 7 hLF-expressing clones were selected with clone 121 exhibiting the highest expression level. The hLF-containing algal extract significantly inhibited the growth of bacteria such as Escherichia coli and Klebsiella variicola. During acute toxicity experiment no acute toxicity was detected, especially on changes of the body weight and histopathology of organs. The recombinant hLF possessed a similar or modestly reduced stability compared to commercial hLF standard. Our data indicated that expression of hLF in C. reinhardtii is feasible and paved a way to commercial production of lactoferrin using edible Chlamydomonas expression system. Abbreviations: atrazine chlorohydrolase gene (atzA); bovine serum albumin (BSA); human LF (hLF); lactoferrin (LF); Luria-Bertani (LB); quantitative reverse transcriptase PCR (qRT-PCR) ; SDS polyacrylamide gel electrophoresis (SDS-PAGE); Tris-acetate phosphate (TAP); western blotting (WB) Expression of human lactoferrin in Chlamydomonas reinhardtii had significant antibacterial activity, stable physical-chemical property and no acute toxicity to mice. [ABSTRACT FROM AUTHOR]

Background: There is a need for a standardized method for quantification of lactoferrin in infant formulas, and manufacturers have started fortifying lactoferrin to mimic the higher levels found in human milk. A variety of current methods exist, but specificity and accuracy are challenging with the infant formula matrix. The use of signature peptides and MS is becoming more prevalent in the realm of analytical chemistry for quantification of proteins. Objective: The objective of this work was to develop and validate a method through a single-laboratory validation for quantification of lactoferrin in milk-based infant formula and begin to lay the foundation for a standardized method. Methods: The method presented uses signature peptides to quantify lactoferrin in milk-based infant formulas by ultra-high performance LC-tandem mass spectrometry (MS/MS). These peptides are produced through tryptic digestion, and fragments produced from these peptides through MS/MS allow the specific quantification using correlating isotopically labeled peptides. Results: The validation parameters were all met with precision RSDr ranging from 2.1 to 7.1 and intermediate RSDR ranging from 7.0 to 10.4 across different fortified milk-based infant formulas. Accuracy with certified reference material resulted in mean recoveries of 91.7–96.4%. Conclusions: The results from this study demonstrate the method is fit for purpose to support manufacturing specifications and nutritional labeling requirements. [ABSTRACT FROM AUTHOR]

Objectives: To evaluate the in vitro antimicrobial/antivirulence action of bovine lactoferrin and its ability to synergize with levofloxacin against resistant Helicobacter pylori strains and to analyse the effect of levofloxacin, amoxicillin and esomeprazole with and without bovine lactoferrin as the first-line treatment for H. pylori infection.Methods: The bovine lactoferrin antimicrobial/antivirulence effect was analysed in vitro by MIC/MBC determination and twitching motility against six clinical H. pylori strains and a reference strain. The synergism was evaluated using the chequerboard assay. The prospective therapeutic trial was carried out on two separate patient groups, one treated with esomeprazole/amoxicillin/levofloxacin and the other with esomeprazole/amoxicillin/levofloxacin/bovine lactoferrin. Treatment outcome was determined with the [13C]urea breath test.Results: In vitro, bovine lactoferrin inhibited the growth of 50% of strains at 10 mg/mL and expressed 50% bactericidal effect at 40 mg/mL. The combination of levofloxacin and bovine lactoferrin displayed a synergistic effect for all strains, with the best MIC reduction of 16- and 32-fold for levofloxacin and bovine lactoferrin, respectively. Bovine lactoferrin at one-fourth MIC reduced microbial motility significantly for all strains studied. In the in vivo study, 6 of 24 patients recruited had treatment failure recorded with esomeprazole/amoxicillin/levofloxacin (75% success, 95% CI 57.68%-92.32%), and in the group with esomeprazole/amoxicillin/levofloxacin/bovine lactoferrin, 2 out of 53 patients recruited had failure recorded (96.07% success, 95% CI 90.62%-101.38%).Conclusions: Bovine lactoferrin can be considered a novel potentiator for restoring susceptibility in resistant H. pylori strains. Bovine lactoferrin added to a triple therapy in first-line treatment potentiates the therapeutic effect. [ABSTRACT FROM AUTHOR]

Captive‐bred rhesus macaques of Indian origin represent one of the most important large animal models for infectious disease, solid organ transplantation, and stem cell research. There is a dearth of information defining hematopoietic development, including neutrophil leukocyte differentiation in this species using multicolor flow cytometry. In the current study, we sought to identify cell surface markers that delineate neutrophil progenitor populations with characteristic immunophenotypes. We defined four different postmitotic populations based on their CD11b and CD87 expression pattern, and further refined their immunophenotypes using CD32, CD64, lactoferrin, and myeloperoxidase as antigenic markers. The four subsets contained myelocyte, metamyelocyte, band, and segmented neutrophil populations. We compared our flow cytometry‐based classification with the classical nuclear morphology‐based classification. We found overlap of immunological phenotype between populations of different nuclear morphology and identified phenotypically different subsets within populations of similar nuclear morphology. We assessed the responsiveness of these populations to stimulatory signals, such as LPS, fMLP, or PMA, and demonstrated significant differences between human and rhesus macaque neutrophil progenitors. In this study, we provided evidence for species‐specific features of granulopoiesis that ultimately manifested in the divergent immunophenotypes of the fully differentiated segmented neutrophils of humans and rhesus macaques. Additionally, we found functional markers that can be used to accurately quantify neutrophil progenitors by flow cytometry. Although these markers do not coincide with the classical nuclear‐morphology‐based grading, they enable us to perform functional studies monitoring immunophenotypic markers. A flow cytometric protocol using species‐specific characteristics to define rhesus macaque neutrophil progenitors in the bone marrow. [ABSTRACT FROM AUTHOR]

Objectives: To determine the antimicrobial activity of ALX-009, a combination of bovine lactoferrin and hypothiocyanite, in sputum against Pseudomonas aeruginosa and Burkholderia cepacia complex (Bcc), key pathogens causing infection in the lungs of cystic fibrosis (CF) patients.Methods: The antimicrobial activity of ALX-009 against clinical respiratory P. aeruginosa isolates was determined by time-kill assay. Sputum from CF patients was treated with ALX-009, either alone or in combination with tobramycin, and the effect on P. aeruginosa, Bcc and total sputum density was determined.Results: Time-kill assay indicated that ALX-009 was bactericidal at 24 h against 4/4 P. aeruginosa isolates under aerobic conditions, and against 3/4 isolates under anaerobic conditions. ALX-009 was also bactericidal against P. aeruginosa in sputum samples at 6 h (n = 22/24 samples) and 24 h (n = 14/24 samples), and demonstrated significantly greater activity than tobramycin at both timepoints. Activity against Bcc in sputum samples (n = 9) was also demonstrated, but the magnitude of change in Bcc density was less than for P. aeruginosa. To determine the effect of treating sputum with two doses of ALX-009, similar to current regimens for inhaled antibiotics, aliquots of a further 10 sputum samples positive for P. aeruginosa were treated with one (t = 0 h) or two doses (t = 0 h, t = 12 h) of ALX-009; treatment with two doses resulted in bactericidal activity in 7/10 samples at 34 h compared with only 3/10 samples when treatment was with one dose.Conclusions: ALX-009 demonstrates promise as a novel antimicrobial that could be used to decrease P. aeruginosa density in the lungs of people with CF. [ABSTRACT FROM AUTHOR]

Exposing Holstein dry cows to short-day photoperiod (SDPP) of 8 hours per day can improve milk production post-calving by 3.2 kg per day, compared with cows exposed to long-day photoperiod (LDPP) of 16 hours per day. Based on seasonal variations in colostrum quality and quantity, we hypothesized that altered photoperiod during the dry period also impacts colostrum production in dairy cows. Our objective was to study the effect of LD and SD during the dry period on colostrum production in Holstein and Jersey cows. Holstein and Jersey cows (n = 33) dried off 60 days before their due date were exposed to either SD (n = 9 Holstein, n = 8 Jersey) or LD (n = 8 Holstein, n = 8 Jersey) photoperiod at 18.3 °C until calving. Within 1 hour of calving, colostrum production was measured (kg), Brix score was determined and a milk sample was collected to evaluate composition (fat, protein, other solids, lactose, lactoferrin, somatic cell count) by DHIA and immunoglobulin content (IgA, IgG, IgG1, IgM) via ELISA. Data were analyzed with the PROC MIXED procedure of SAS 9.4 with fixed effects of breed and photoperiod treatment. Replicate, animal age, number of days of treatment, season of calving, inbreeding percentage and previous lactation production were included as random effects. Only the percentage of IgG made up by IgG1 differed by treatment, being greater for SDPP cows (79.6% ± 4.6%) than LDPP cows (68.0% ± 3.1%; P < 0.05). Jerseys had greater Brix score (27.8% ± 0.7% vs. 23.4% ± 0.7%), colostral protein (9.3% ± 0.3% vs. 7.6% ± 0.3%) and IgA (5.7 ± 0.7 vs. 3.2 ± 0.5 g/L; P < 0.01), and a tendency for fat and IgM to be greater as well (P < 0.10). These data indicate that measures of colostrum quantity and quality differ by breed but are largely unaffected by photoperiod treatment. [ABSTRACT FROM AUTHOR]

Abstract: Ascitic neutrophils from cirrhotic patients with spontaneous bacterial peritonitis (SBP) exhibit an impaired oxidative burst that could facilitate bacterial infection. However, the influence of the cell‐free ascitic fluid of these patients on neutrophil function has not been investigated. To analyze this influence, we determined the ascitic levels of cytokines, resistin, and lactoferrin and their association with neutrophil function, disease severity score, and SBP resolution. We analyzed NETosis induction by microscopy and oxidative burst by the flow cytometry of healthy neutrophils cultured in ascitic fluid from cirrhotic patients with sterile ascites (SA) and with SBP before and after antibiotic treatment. Resistin, IL‐6, IL‐1 receptor antagonist, IL‐1β, and lactoferrin levels were measured in ascitic fluids and supernatants of cultured neutrophils and PBMCs by ELISA. Upon stimulation, healthy neutrophils cultured in SBP ascitic fluid produced lower NETosis and oxidative burst than those cultured in SA. Ascitic resistin levels were negatively correlated with NETosis, oxidative burst, and ascitic glucose levels; and positively correlated with the model for end‐stage liver disease score. After an E. coli or TNF‐α stimulus, neutrophils were the major resistin producers. Resistin indirectly reduced the oxidative burst of neutrophils and directly reduced the inflammatory phenotype of monocytes and TNF‐α production. Bacterial‐induced resistin production can down‐regulate the inflammatory response of macrophages and neutrophil function in ascitic fluid. Consequently, this down‐regulation may jeopardize the elimination of bacteria that translocate to ascitic fluid in patients with cirrhosis. [ABSTRACT FROM AUTHOR]

Summary: Bioactive components of human milk, such as human lactoferrin (hLF), play an essential role in gut microbiome homeostasis and protection against neonatal inflammatory diseases. Neonatal intestinal macrophages display a proinflammatory profile that might contribute to inflammatory mucosal injury. Therefore, the aim of the study was to investigate the immunomodulatory effects of hLF on differentiation and activation of monocyte‐derived macrophages (moMϕ). Monocytes isolated from umbilical cord blood of term neonates and peripheral blood of healthy adults were differentiated in the absence or presence of hLF, and differentiation, apoptosis and phagocytosis were evaluated. Cytokine production, Toll‐like receptor (TLR) signalling and activation marker expression were investigated upon activation with lipopolysaccharide (LPS) and lipoteichoic acid (LTA) challenge. We demonstrate that hLF‐differentiated moMϕ exhibit decreased TLR‐4 expression, TLR signalling, proinflammatory cytokine secretion and intracellular tumour necrosis factor (TNF)‐α production. Investigation of differentiation markers, morphology and induction of apoptosis showed no alteration in lactoferrin‐differentiated moMϕ. Taken together, hLF promote anergic/anti‐inflammatory effects by TLR expression and pathway interference, resulting in a diminished proinflammatory moMϕ phenotype. The anergic/anti‐inflammatory properties of hLF might contribute to the prevention of harmful TLR‐mediated inflammatory disorders in the developing gut of premature infants. [ABSTRACT FROM AUTHOR]

This study aimed to establish an event-specific multiplex PCR system using microsatellite markers and fluorescently labeled primers to detect six different genetically modified (GM) animal lines, including human lactoferrin GM cattle, human lysozyme GM cattle, human α-lactalbumin GM cattle, myostatin knockout pigs, phytase GM pigs, and ω-3 fatty acid desaturase gene GM pigs. Four different microsatellite loci for species identification, along with six GM animal-specific fragments, were selected as targets for primer design. The capillary gel electrophoresis results of multiplex PCR showed that the target fragments were amplified successfully. This high-throughput multiplex PCR detection system can be applied for the inspection and quarantine of GM animals. [ABSTRACT FROM AUTHOR]

Background and Aims: Inflammatory bowel disease [IBD] is characterised by a disruption of immune homeostasis, which is tightly regulated to protect against harmful pathogens yet not react to commensal antigens. Animal studies indicate that regulatory T cells [Treg] modulate the immune response to prevent IBD development. Lactoferrin [LF] is an endogenous anti-inflammatory pleiotropic protein secreted at high concentrations in colostrum and at mucosal sites. However, the effect of LF on specific T lymphocyte populations has not been studied. Here, we identify a novel mechanism by which a recombinant human LF, VEN-120, regulates T cell populations in health and disease. Methods: Two murine models of intestinal inflammation, the dextran sodium sulphate colitis model and the TNFΔARE/+ model of ileitis, were used to study the anti-inflammatory and T cell modulating ability of VEN-120. Flow cytometry was used to evaluate T cell populations within the lamina propria and mesenteric lymph nodes, and to evaluate the effect of VEN-120 on CD4+ T cells in vitro. Results: VEN-120 reduced inflammation in both models of IBD, accompanied by increased Tregs in the intestinal lamina propria. Treatment of CD4+ T cells in vitro resulted in an upregulation of Treg genes and skewing towards a Treg population. This in vitro T cell skewing translated to an increase of Treg homing to the intestinal lamina propria and associated lymph tissue in healthy mice. Conclusions: These data provide a novel immunological mechanism by which VEN-120 modulates T cells to restrict inflammatory T cell-driven disease. [ABSTRACT FROM AUTHOR]

The pathogenic Neisseria provide textbook examples of phase variation: the high frequency, random and reversible switching of gene expression. Typically, phase variable gene expression is observed in genes required for the expression of surface proteins and carbohydrate structures. All Neisseria gonorrhoeae and N. meningitidis strains also express phase variable DNA methyltransferases that are components of DNA restriction-modification systems. Phase variation of these DNA methyltransferases (Mod) alters global DNA methylation patterns. The change in DNA methylation due to phase variation events alters expression of a regulon of genes, called a phasevarion, and results in differentiation of the population into cells with two distinct phenotypes. For example, in N. meningitidis switching of the modA11 phasevarion alters expression of immunogenic outer membrane proteins such as lactoferrin-binding protein, and also modulates sensitivity to ceftazidime and ciprofloxacin. The modD1 phasevarion is associated with hypervirulent meningococcal clonal complexes. In N. gonorrhoeae, modA13 phasevarion switching generates differentiation into cells that display enhanced biofilm formation and enhanced intracellular survival. Phasevarions are ubiquitous in pathogenic Neisseria and modulate expression of numerous genes. These systems have the potential to impact all studies on vaccine development and pathobiology in the pathogenic Neisseria. [ABSTRACT FROM AUTHOR]

Background: Faecal calprotectin is considered to be a valid test for ruling out inflammatory bowel disease (IBD) in children with chronic gastrointestinal symptoms in specialist care. In contrast, faecal lactoferrin has higher specificity. The recent availability of both as point-of-care tests (POCTs) makes them attractive for use in primary care.Objective: To evaluate the test characteristics of calprotectin and lactoferrin POCTs for diagnosing IBD in symptomatic children.Methods: We defined two prospective cohorts of children with chronic gastrointestinal symptoms: (i) children presenting to primary care (primary care cohort); (ii) children referred for specialist care (referred cohort). Baseline POCT results were compared with the outcome of either endoscopic assessment or 12 months follow-up. Clinicians were blinded to the POCT results.Results: In the primary care cohort, none of the 114 children had IBD, and the calprotectin and lactoferrin POCTs had specificities of 0.95 (0.89-0.98) and 0.98 (0.93-0.99), respectively. In the referred cohort, 17 of the 90 children had IBD: the sensitivity of POCT calprotectin and POCT lactoferrin were both 0.94 (0.72-0.99); and the specificity was 0.93 (0.84-0.97) and 0.99 (0.92-1.00), respectively. The POCT calprotectin could reduce the referral rate by 76% and POCT lactoferrin by 81%, while missing one child with IBD (6%).Conclusion: A diagnostic test strategy in primary care using a simple POCT calprotectin or lactoferrin has the potential to reduce the need for referral for further diagnostic work-up in specialist care, with a low risk of missing a child with IBD. [ABSTRACT FROM AUTHOR]

Central nervous system (CNS) relapse is a devastating complication that occurs in about 5% of diffuse large B-cell lymphoma (DLBCL) patients. Currently, there are no predictive biological markers. We wanted to study potential biomarkers of CNS tropism that play a role in adhesion, migration and/or in the regulation of inflammatory responses. The expression levels of ITGA10, CD44, PTEN, cadherin-11, CDH12, N-cadherin, P-cadherin, lactoferrin and E-cadherin were studied with IHC and IEM. GEP was performed to see whether found expressional changes are regulated at DNA/RNA level. IHC included 96 samples of primary CNS lymphoma (PCNSL), secondary CNS lymphoma (sCNSL) and systemic DLBCL (sDLBCL). IEM included two PCNSL, one sCNSL, one sDLBCL and one reactive lymph node samples. GEP was performed on two DLBCL samples, one with and one without CNS relapse. CNS disease was associated with enhanced expression of cytoplasmic and membranous ITGA10 and nuclear PTEN (P < 0.0005, P = 0.002, P = 0.024, respectively). sCNSL presented decreased membranous CD44 and nuclear and cytoplasmic cadherin-11 expressions (P = 0.001, P = 0.006, P = 0.048, respectively). In PCNSL lactoferrin expression was upregulated (P < 0.0005). IEM results were mainly supportive of the IHC results. In GEP CD44, cadherin-11, lactoferrin and E-cadherin were under-expressed in CNS disease. Our results are in line with previous studies, where gene expressions in extracellular matrix and adhesion-related pathways are altered in CNS lymphoma. This study gives new information on the DLBCL CNS tropism. If further verified, these markers might become useful in predicting CNS relapses. [ABSTRACT FROM AUTHOR]

Lactoferrin (LF) is produced by exocrine glands including salivary gland, and has various functions including infection defense. However, the transfer of LF from peripheral organs into the brain remains unclear. To clarify the kinetics of salivary LF (sLF), we investigated the consequences of sialoadenectomy and bovine LF (bLF) sublingual administration in rats. The salivary glands were removed from male Wistar rats, and we measured rat LF levels in the blood and brain at 1 week post-surgery. We also examined the transfer of LF into the organs of the rats after sublingual administration of bLF. Rat LF levels in the blood and brain were significantly reduced by sialoadenectomy. Sublingual bLF administration significantly increased bLF levels in the brain, which then decreased over time. These results indicate that LF is transferred from the sublingual mucosa to the brain, in which favorable effects of sLF on brain will be expected via the sublingual mucosa. LF levels in the brain significantly decreased by sialoadenectomy (A). LF was also transferred from the sublingual mucosa to the brain (B). [ABSTRACT FROM PUBLISHER]

In the cervicovaginal microenvironment, lactobacilli are known to protect against genital infections and, amongst the host defence compounds, lactoferrin has recently acquired importance for its anti-microbial and anti-inflammatory properties. An abnormal genital microenvironment facilitates the acquisition of pathogens like Chlamydia trachomatis, the leading cause of bacterial sexually transmitted infections worldwide. The aim of our study is to investigate the effects of Lactobacillus crispatus, Lactobacillus brevis and bovine lactoferrin on chlamydial infection, in order to shed light on the complex interplay between host defence mechanisms and C. trachomatis. We have also evaluated the effect of these defence factors to modulate the chlamydia-mediated inflammatory state. To this purpose, we have determined the infectivity and progeny production of C. trachomatis as well as interleukin-8 and interleukin-6 synthesis. The main result of our study is that the combination of L. brevis and bovine lactoferrin is the most effective in inhibiting the early phases (adhesion and invasion) of C. trachomatis infection of cervical epithelial cells and in decreasing the levels of both cytokines. In conclusion, the interaction between L. brevis and lactoferrin seems to play a role in the protection against C. trachomatis, reducing the infection and regulating the immunomodulatory activity, thus decreasing the risk of severe complications. [ABSTRACT FROM AUTHOR]

To better understand underlying causes of lower rotavirus vaccine effectiveness in low-middle income countries (LMICs), we measured innate antiviral factors in Nicaraguan mothers' milk and immune response to the first dose of the pentavalent rotavirus vaccine in corresponding infants. No relationship was found between concentrations of innate factors and rotavirus vaccine response. [ABSTRACT FROM AUTHOR]

Background: Moraxella catarrhalis is an established pathogen that is causing substantial infections to both children and adults. However, so far there is no effective vaccine to halt the spread of these infections.Methods: Immunoinformatics tools were used to predict M. catarrhalis epitopes that could offer immunoprotection among major proportions of human populations worldwide. Mice were immunized with the best 3 peptides and then challenged with M. catarrhalis in the pulmonary clearance model. Finally, antibodies against these epitopes were detected in humans.Results: Immunoinformatics analyses identified 44 epitopes that are predicted to be good major histocompatibility complex class II binders and at the same time show high population coverage worldwide. After intraperitoneal immunization of mice with the best 3 peptides, peptide A, derived from lactoferrin-binding protein A, showed superior activity in immunogenicity and in clearing M. catarrhalis from mouse lungs. Higher clearance was obtained by combining intraperitoneal and intranasal immunization. In the serum samples from children with otitis media infected with M. catarrhalis, antibody levels against peptide A were significantly lower than in samples from children without otitis media.Conclusions: Peptide A is the first promising peptide-based vaccine against M. catarrhalis Immunoinformatics predicts that it should have a global protection around the world. [ABSTRACT FROM AUTHOR]

Objectives: Clostridium difficile infection (CDI) is a global healthcare problem. Recent evidence suggests that the availability of iron may be important for C. difficile growth. This study evaluated the comparative effects of iron-depleted (1% Fe(3+) saturated) bovine apo-lactoferrin (apo-bLf) and iron-saturated (85% Fe(3+) saturated) bovine holo-lactoferrin (holo-bLf) in a human in vitro gut model that simulates CDI.Methods: Two parallel triple-stage chemostat gut models were inoculated with pooled human faeces and spiked with C. difficile spores (strain 027 210, PCR ribotype 027). Holo- or apo-bLf was instilled (5 mg/mL, once daily) for 35 days. After 7 days, clindamycin was instilled (33.9 mg/L, four times daily) to induce simulated CDI. Indigenous microflora populations, C. difficile total counts and spores, cytotoxin titres, short chain fatty acid concentrations, biometal concentrations, lactoferrin concentration and iron content of lactoferrin were monitored daily.Results: In the apo-bLf model, germination of C. difficile spores occurred 6 days post instillation of clindamycin, followed by rapid vegetative cell proliferation and detectable toxin production. By contrast, in the holo-bLf model, only a modest vegetative cell population was observed until 16 days post antibiotic administration. Notably, no toxin was detected in this model. In separate batch culture experiments, holo-bLf prevented C. difficile vegetative cell growth and toxin production, whereas apo-bLf and iron alone did not.Conclusions: Holo-bLf, but not apo-bLf, delayed C. difficile growth and prevented toxin production in a human gut model of CDI. This inhibitory effect may be iron independent. These observations suggest that bLf in its iron-saturated state could be used as a novel preventative or treatment strategy for CDI. [ABSTRACT FROM AUTHOR]

Previously, we found that enteric lactoferrin (eLF) could reduce the visceral fat accumulation known to associate strongly with metabolic syndrome symptoms and consequently with an increased risk of atherosclerosis. In this study, the atherosclerosis-preventive potential of LF was assessed in a high-fat and high-cholesterol diet (HFCD)-induced hypercholesterolemia and atherosclerosis model using Microminipig™. Eight-week orally administered eLF remarkably reduced the HFCD-induced serum total and low-density lipoprotein cholesterol levels but not high-density lipoprotein cholesterol levels. A histological analysis of 15 arteries revealed that eLF systemically inhibited the development of atherosclerotic lesions. Pathway analysis using identified genes that characterized eLF administration in liver revealed significant changes in the steroid biosynthesis pathway (ssc00100) and all affected genes in this pathway were upregulated, suggesting that cholesterol synthesis inhibited by HFCD was recovered by eLF. In summary, eLF could potentially prevent the hypercholesterolemia and atherosclerosis through protecting homeostasis from HFCD-induced dysfunction of cholesterol metabolism. [ABSTRACT FROM PUBLISHER]

Lactoferrin (LTF), a multifunctional glycoprotein of the transferrin family mainly found in exotic secretions in mammals, is an important defense molecule against not only microbial invasion but also tumors. It folds into two globular domains (N- and C-lobes) each containing an iron-binding site. The cationic antimicrobial peptide in N-lobe is known to exert anti-tumor effect via a non-receptor-mediated pathway. However, whether LTF C-lobe also contributes to its anti-tumor activity remains to be investigated. In this study, a human LTF fragment (amino acid residues 343–682) covering the C-lobe was expressed with a histidine tag inE. coliand the purified polypeptide refolded through a series of buffer changing procedure. The resultant recombinant protein caused significant growth arrest of breast carcinoma cells MDA-MB-231 in a dose- and time-dependent manner, evidently via induction of apoptosis of the cell. Our data suggest a positive role for the C-lobe of human LTF in controlling tumors in vitro. Recombinant human lactoferrin C-lobe exhibited significant inhibitory effect on the growth of MDA-MB-231 cells and morphological changes in a dose and time-dependent manner. [ABSTRACT FROM PUBLISHER]

The objectives of this study were to determine if newborn calves receiving supplemental lactoferrin (LF) had improved IgG uptake and if supplemental LF enhanced intestinal development through estimation of xylose uptake. Twenty-four newborn Holstein bull calves were randomly assigned to 1 of 2 treatments: 0 or 1 g/d of supplemental LF. Calves were fed pooled maternal colostrum from 9 cows in 2 feedings: at birth and 12 h later. Calves consumed in excess of 200 g of IgG. Blood samples were taken before colostrum feeding (0 h) and at 12, 18, and 24 h after birth. Blood samples were analyzed for IgG concentration. On d 2 of life, calves were fed milk replacer with the added LF and 0.5 g/kg of BW xylose to determine if supplemental LF affected intestinal development. Blood was sampled at 0, 0.5, 1, 2, 3, 4, 6, 8, and 12 h after the xylose dose. All calves attained passive transfer and supplemental LF did not affect IgG uptake (P ≥ 0.36) or apparent efficiency of absorption of IgG (P = 0.49). Lactoferrin did not enhance rate of absorption at any time point (P ≥ 0.36). There were no differences in xylose (P = 0.28) or glucose (P = 0.27) area under the curve values in calves supplemented with either 0 or 1 g/d LF. Lactoferrin did not enhance IgG uptake during the first 24 h or intestinal development in calves on the second day of life. [ABSTRACT FROM AUTHOR]

Infant formula is designed to provide the human infant with a sole source of nutrition and it is intended to imitate breast milk. In recent years, advances in the science of infant nutrition have led to an increasing number of novel ingredients that are supplemented into infant formula. As the list of these nutritionally important nutrients is lengthy, this review summarizes contemporary analytical methods that have been applied to a representative selection (lutein, carnitine, choline, nucleotides, inositol, taurine, sialic acid, gangliosides, triacylglycerides, oligosaccharides, α-lactalbumin, and lactoferrin). [ABSTRACT FROM AUTHOR]

Intake of poor-quality colostrum reduces thriftiness and survival of dairy calves and cost dairy producers in 2017 $70 per head for each calf receiving poor quality colostrum. Studies suggest levels of lactoferrin and IgA within good colostrum are 1.5 - 5 mg/ml and 5.36 mg/ml respectively. Today, producers are recommended to expose cows to short-day photoperiod (SDPP) during the dry period as this has been shown to increase milk production during the next lactation. However, the impact of SDPP on colostrum quality is unknown. The objective of this study was to determine the effect of day length during the entire dry period on the colostrum quality. We measured the levels of lactoferrin and IgA from both Jersey and Holstein dry cows (n=33) exposed to 8 hours of light for SDPP (n=9 Holstein, n=8 Jersey) or 16 hours of light for long-day photoperiod (LDPP) (n=8 Holstein, n=8 Jersey) using enzyme-linked immunosorbent assays (Bethyl Laboratories). Colostrum samples were collected from the cows, skimmed and frozen at -20oC until assayed. Data were analyzed with SAS 9.4. We found that the levels of lactoferrin and IgA did not differ between breed and photoperiod treatment (P = 0.57, P = 0.46, respectively). The average for lactoferrin in SDPP animals was 1.41 mg/mL and 1.55 mg/mL for LDPP. The average for IgA in SDPP animals was 5.08 mg/mL and 3.88 mg/mL for LDPP. Based on these data we suggest that dry period photoperiod exposure does not impact colostral IgA or lactoferrin levels. [ABSTRACT FROM AUTHOR]

The objective of this study was to determine if feed restriction (FR) followed by feeding a high concentrate (HC) diet would induce leaky gut syndrome (LGS) in cattle. Changes in circulatory and fecal biomarkers were utilized to assess LGS. Individually penned Holstein heifers (n=16; BW=343.2 ± 3.2 kg) were used in a completely randomized design with animal as the experimental unit. Cattle were fed a forage-based diet (71.0% hay, 12.0% cracked corn, 17.0% DDGS) twice daily ad libitum from day 0-14; from day 15-17, the daily ration was restricted by 60%. One of two HC diets (n=8 each) were fed ad libitum from day 18-30: HC75=17.0% hay, 70.0% cracked corn, 13.0% DDGS; HC65=26.5% hay, 60.0% cracked corn, 13.5% DDGS. Blood and fecal samples were collected prior to a.m. feed delivery (d -1, 0, 14, 15, 18, 20, 24, 30) and analyzed for serum amyloid-A (SAA), haptoglobin (Hp), lipopolysaccharide-binding protein (LBP), fecal lactoferrin, and fecal pH. Variables were analyzed using a mixed model with repeated measures; study day, treatment and the interaction were fixed effects. Following FR, SAA was significantly increased from day 14 to 24 (27,406.20 ± 8,917.00 ng/mL; P = 0.046). Similarly, LBP was significantly increased from day 14 to 20 (353.71 ± 44.40 ng/mL; P < 0.0001). Fecal lactoferrin increased significantly from day 18 to 20 (293.9 ± 57.54 ng/g; P = 0.0001). Fecal pH decreased significantly (1.48 ± 0.09; P < 0.0001) from day 14 to 30 and from day 18 to 30 (1.57 ± 0.08; P < 0.0001) reaching its nadir on day 30 (5.35). No meaningful changes were observed for Hp. Based on these data, FR followed by feeding a HC diet effectively experimentally induced LGS as indicated by observed changes in SAA, LBP, fecal lactoferrin and fecal pH. [ABSTRACT FROM AUTHOR]

Background: Sepsis is a clinical syndrome caused by many infections. Septic patients may suffer from Gut failure resulting in severe complications. To improve Gut function, the effect of lactofferin supplementation has been studied. Objective: To test the effect of lactoferrin supplementation on improving gut barrier function by using plasma citrulline level as a marker of functional enterocytic mass. Patients and Methods: This double armed single blinded therapeutic clinical trial study was conducted on 40 children admitted to (PICUs) of Ain Shams University Hospitals. By diagnosis sepsis and gut failure, twenty of them weren’t supplemented with LF (group 1) and the other twenty were supplemented (group 2). All critically ill septic patients diagnosed with acute gut failure admitted from October 2018 to 2019 were subjected to citrulline level measurement by (ELISA). Results: We found that citrulline concentration was significantly lower in patients (9.52 nmol/L) than control group (122 nmol/L) and lowered in non survivors (5.13 nmol/L) than survivors (30.68 nmol/L) with p value of 0.027 and 0.034 respectively and a cut off value ≤ 7.82 mmol/L. There was no significant difference between the groups 1 and 2 in treatment of sepsis with gut failure. Conclusion: (1) Citrulline level can be used as a predictor of intestinal failure severity and mortality. (2) Lactoferrin supplementation was not beneficial in pediatric septic patient with gut failure and did not improve outcome or prognosis of patients. [ABSTRACT FROM AUTHOR]

Aims To determine the role of human lactoferrin ( hLF) in protecting the oral cavities of mice against Candida albicans infection in lactoferrin knockout ( LFKO−/−) mice was compared to wild-type ( WT) mice. We also aim to determine the protective role of hLF in LFKO−/− mice. Methods and Results Antibiotic-treated immunosuppressed mice were inoculated with C. albicans (or sham infection) by oral swab and evaluated for the severity of infection after 7 days of infection. To determine the protective role of hLF, we added 0·3% solution of hLF to the drinking water given to some of the mice. CFU count, scoring of lesions and microscopic observations were carried out to determine the severity of infection. LFKO−/−I mice showed a 2 log ( P = 0·001) higher CFUs of C. albicans in the oral cavity compared to the WT mice infected with C. albicans ( WTI). LFKO−/−I mice given hLF had a 3 log ( P = 0·001) reduction in CFUs in the oral cavity compared to untreated LFKO−/−I mice. The severity of infection, observed by light microscopy, revealed that the tongue of the LFKO−/−I mice showed more white patches compared to WTI and LFKO−/−I + hLF mice. Scanning electron microscopic observations revealed that more filiform papillae were destroyed in LFKO−/−I mice when compared to WTI or LFKO−/−I + hLF mice. Conclusions Human LF is important in protecting mice from oral C. albicans infection. Administered hLF may be used to prevent C. albicans infection. Significance and Impact of the Study Human LF, a multifunctional iron-binding glycoprotein can be used as a therapeutic active ingredient in oral healthcare products against C. albicans. [ABSTRACT FROM AUTHOR]

Lactoferrin has been widely studied over the last 70 years, and its role in diverse biological functions is now well known and generally accepted by the scientific community. Usually, alterations of the lactoferrin gene in cells are associated with an increased incidence of cancer. Several studies suggest that exogenous treatment with lactoferrin and its derivatives can efficiently inhibit the growth of tumors and reduce susceptibility to cancer. None of these studies, however, reported a consistent outcome with regard to the mechanisms underlying the anticancer effects of lactoferrin. In this review, the association of lactoferrin with cancer is thoroughly discussed, from lactoferrin gene expression to the potential use of lactoferrin in cancer therapy. Lactoferrin cytotoxicity against several cancers is reported to occur in distinct ways under different conditions, namely by cell membrane disruption, apoptosis induction, cell cycle arrest, and cell immunoreaction. Based on these mechanisms, new strategies to improve the anticancer effects of the lactoferrin protein and/or its derivatives are proposed. The potential for lactoferrin in the field of cancer research (including as a chemotherapeutic agent in cancer therapy) is also discussed. [ABSTRACT FROM AUTHOR]

The article focuses on a cross-linking between psoriasis, inflammation and renal function impairment as moderate-to-severe psoriasis presents an increased risk chronic kidney disease (CKD). Topics discussed include psoriasis may cause hypertension and diabetes, evaluation of relationship between serum creatinine and some inflammatory markers and Interleukin(IL)-6, tumour necrosis factor (TNF)-alpha and and lactoferrin evaluated by enzyme immunoassay.

Recent advances in perinatal and neonatal intensive care have resulted in significant improvements in the survival of preterm extremely low-birthweight ( PELBW) infants; however, extrauterine growth restriction ( EUGR) and undernutrition occur frequently during hospitalization and are associated with adverse outcomes, including bronchopulmonary dysplasia, sepsis, and neurodevelopmental impairment. Early optimal parenteral nutrition with adequate amino acids and lipids, especially long-chain polyunsaturated fatty acids, has been shown to decrease the incidence of EUGR, bronchopulmonary dysplasia, necrotizing enterocolitis, sepsis, and retinopathy of prematurity in animal models and clinical trials. In PELBW infants, breast milk and probiotics have been shown to reduce the incidence of necrotizing enterocolitis, and lactoferrin has been demonstrated to prevent late-onset sepsis. Thus, early administration of optimal postnatal parenteral and enteral nutrients can help prevent neurodevelopmental impairment caused by EUGR, necrotizing enterocolitis, sepsis, bronchopulmonary dysplasia, and retinopathy of prematurity, and recent evidence indicates such treatment is feasible. [ABSTRACT FROM AUTHOR]

ABSTRACT Lactoferrin (LF) has been established as a potent anabolic factor for bone health both in vivo and in vitro. However, the molecular mechanisms underlying LF's action are still largely unknown. Here, we explore the signaling pathways that mediate LF's beneficial effect on osteoblast differentiation. In primary osteoblast and preosteoblast MC3T3-E1, LF promoted alkaline phosphatase (ALP) activity, osteocalcin (OCN) secretion, and mineralization. Along with this enhanced osteogenic differentiation, activation of p38 mitogen-activated protein kinase (MAPK) was detected in LF-treated MC3T3-E1 cells. Downregulating p38 with selective inhibitor SB203580 or p38α small interfering RNA (siRNA) attenuated the effect of LF on osteogenesis. Furthermore, knockdown of p38α significantly decreased LF-induced Runt-related transcription factor 2 (Runx2) phosphorylation. According to previous studies and our results, we speculated that LF-induced osteoblast proliferation and differentiation were two relatively separate processes controlled by extracellular signal-regulated kinase 1/2 (ERK1/2) and p38 pathways, respectively. Besides p38 MAPK activation, protein kinase A (PKA) was also activated in MC3T3-E1 cells. PKA inhibitor H89 significantly inhibited LF-induced p38 activation, ALP activity, and OCN secretion, indicating that PKA possibly acted as an upstream kinase of p38. In order to further identify the role of LF's receptor low-density lipoprotein receptor-related protein 1 (LRP1), we constructed LRP1 stable-knockdown MC3T3-E1 cells. Neither LRP1 antagonist receptor associated protein (RAP), nor LRP1 knockdown approach could attenuate the LF-induced osteogenesis, implying that LF stimulated osteoblast differentiation via an LRP1-independent pathway. Taken together, the present work indicated that LF stimulated MC3T3-E1 preosteoblast differentiation mainly through LRP1-independent PKA and p38 signaling pathways. These results provided the first evidence of the signaling mechanisms of LF's effect on osteoblast differentiation. © 2014 American Society for Bone and Mineral Research. [ABSTRACT FROM AUTHOR]

Most secreted and cell membrane proteins in mammals are glycosylated. Many of these glycoproteins are also prevalent in milk and play key roles in the biomodulatory properties of milk and ultimately in determining milk's nutritional quality. Although a significant amount of information exists on the types and roles of free oligosaccharides in milk, very little is known about the glycans associated with milk glycoproteins, in particular, the biological properties that are linked to their presence. The main glycoproteins found in bovine milk are lactoferrin, the immunoglobulins, glycomacropeptide, a glycopeptide derived from κ-casein, and the glycoproteins of the milk fat globule membrane. Here, we review the glycoproteins present in bovine milk, the information currently available on their glycosylation and the biological significance of their oligosaccharide chains. [ABSTRACT FROM AUTHOR]