Transferrin Family Research Articles

Regulation of LPS-Induced Inflammatory Responses in Bovine Mammary Epithelial Cells via TLR4-Mediated NF-κB and MAPK Signaling Pathways by Lactoferrin.

Lactoferrin (LF), a member of the transferrin family, is widely present in mammalian milk and other secretions, exhibiting anti-inflammatory, antibacterial, and anti-infective properties. Although the biological functions of LF have been extensively studied, there are few reports on its effects and molecular mechanisms concerning bovine mastitis caused by bacterial infection. This study used bovine mammary epithelial cells (BMECs) cultured in vitro as the research model. An inflammatory injury model was established by stimulating BMECs with LPS to investigate whether LF at different concentrations (10, 50, 100, and 200 μg·mL-1) could inhibit the inflammatory response before and after the onset of inflammation. The expression of inflammatory cytokines IL-1β, IL-6, IL-8, and TNF-α at both the gene and protein levels was detected using RT-qPCR and ELISA. Western blotting was employed to evaluate the phosphorylation levels in the inflammatory signaling pathways MAPK/P38/ERK and NF-κB/P65, while RT-qPCR was used to examine the impact on TLR4 receptor gene expression. The results display that pretreatment with LF prior to LPS-induced inflammation in BMECs reduced the expression of inflammatory cytokines IL-1β, IL-6, IL-8, and TNF-α at both the gene and protein levels (p < 0.05). LF also inhibited the phosphorylation of NF-κB/P65 and MAPK/P38/ERK signaling pathways and downregulated TLR4 receptor gene expression (p < 0.05). However, when LF was added after the onset of LPS-induced inflammation, inflammatory cytokine expression and phosphorylation levels in the NF-κB/P65 and MAPK/P38/ERK pathways remained elevated, along with high expression of the TLR4 receptor gene (p < 0.05). These findings show that LF can antagonize LPS-induced inflammatory responses in BMECs and reduce cytokine expression, exhibiting anti-inflammatory effects when administered before inflammation. Conversely, when LF is added post-inflammation, it appears to enhance cytokine expression, potentially promoting the recruitment of more cells or factors to resolve inflammation rapidly. Both effects are mediated through the TLR4 receptor and the NF-κB/P65 and MAPK/P38/ERK signaling pathways.

Impact of ovotransferrin on the membrane integrity of Salmonella Enteritidis under egg-white conditions.

Eggs can mediate foodborne disease resulting in salmonellosis outbreaks that are most commonly caused by Salmonella enterica serovar Enteritidis. Ovotransferrin is a prominent egg-white antimicrobial glycoprotein belonging to the transferrin family, members of which exhibit powerful iron-chelating activity. However, several studies have also described the ability of transferrin proteins to disrupt bacterial membranes. This study aimed to investigate the antimicrobial activity of ovotransferrin toward S. Enteritidis membranes at 30°C under egg-white conditions. This aim was supported by the deployment of a synthetic medium designed to mimic egg-white (matching the ionic composition and pH). The ability of ovotransferrin to induce bacterial membrane permeabilization in S. Enteritidis was investigated by measuring substrate accessibility to periplasmic β-lactamase and cytosolic β-galactosidase. The depolarization of the inner membrane of S. Enteritidis was measured using a fluorescence probe [DiSC3(5)]. The results show that ovotransferrin induces permeabilization of the outer membrane but not the inner membrane whereas egg white permeabilizes both membranes. In addition, the dissipation of the proton motive force by egg white was found to involve a contribution by ovotransferrin since this protein provoked inner-membrane depolarization. It can thus be concluded that ovotransferrin exerts a membranes perturbation activity on S. Enteritidis under egg-white conditions, in addition to its well-known iron-chelation activity.

Oral administration of egg white ovotransferrin prevents osteoporosis in ovariectomized rats

Ovotransferrin, an iron-binding glycoprotein, accounting for approximately 12 % of egg white protein, is a member of transferrin family. Our previous studies showed that ovotransferrin stimulates the proliferation and differentiation of osteoblasts, while inhibits osteoclastogenesis and resorption activity. The work aims to study the efficacy of orally administered ovotransferrin on the prevention of osteoporosis using ovariectomized (OVX) Sprague-Dawley rats. Oral administration of ovotransferrin showed no negative effect on body weight, food intake and organ weight. After 12-week treatment, feeding ovotransferrin at a dose of 1 % (1 g ovotransferrin/100 g diet) prevented OVX-induced bone loss and maintained relatively high bone mineral density and integrated bone microarchitecture. The serum concentration of biomarkers indicating bone formation was increased in ovotransferrin administration groups, while the bone resorption biomarkers were decreased. Ovotransferrin feeding also decreased the production of serum cytokine TNF-α and IL-6, which are two stimulators for osteoclast differentiation. In addition to its direct regulatory role on bone turnover, ovotransferrin supplementation might benefit osteoporosis prevention by inhibiting adipogenesis, and regulating immune response. Our results suggested the potential application of ovotransferrin as a functional food ingredient on the prevention of osteoporosis.

The Iron Binding Ability Maps the Fate of Food-Derived Transferrins: A Review.

As the demand for lactoferrin increases, the search for cost-effective alternative proteins becomes increasingly important. Attention naturally turns to other members of the transferrin family such as ovotransferrin. The iron-binding abilities of these proteins influence their characteristics, although the underlying mechanisms remain unclear. This overview systematically summarizes the effects of the iron-binding ability on the fate of food-derived transferrins (lactoferrin and ovotransferrin) and their potential applications. The findings indicate that iron-binding ability significantly influences the structure of food-derived transferrins, particularly their tertiary structure. Changes in structure influence their physicochemical properties, which, in turn, lead to different behaviors in response to environmental variations. Thus, these proteins exhibit distinct digestive characteristics by the time they reach the small intestine, ultimately performing varied physiological functions in vivo. Consequently, food-derived transferrins with different iron-binding states may find diverse applications. Understanding this capability is essential for developing food-derived transferrins and driving innovation in lactoferrin-related industries.

Construction of Iron-Scavenging Hydrogel via Thiol-Ene Click Chemistry for Antibiotic-Free Treatment of Bacterial Wound Infection.

Bacteria, especially drug-resistant strains, can quickly cause wound infections, leading to delayed healing and fatal risk in clinics. With the growing need for alternative antibacterial approaches that rely less on antibiotics or eliminate their use altogether, a novel antibacterial hydrogel named Ovtgel is developed. Ovtgel is formulated by chemically crosslinking thiol-modified ovotransferrin (Ovt), a member of the transferrin family found in egg white, with olefin-modified agarose through thiol-ene click chemistry. Ovt is designed to sequester ferric ions essential for bacterial survival and protect wound tissues from damages caused by the reactive oxygen species (ROS) generated in Fenton reactions. Experimental data have shown that Ovtgel significantly enhances wound healing by inhibiting bacterial growth and shielding tissues from ROS-induced harms. Unlike traditional antibiotics, Ovtgel targets essential trace elements required for bacterial survival in the host environment, preventing the development of drug resistance in pathogenic bacteria. Ovtgel exhibits excellent biocompatibility due to the homology of Ovt to mammalian transferrin. This hydrogel has the potential to serve as an effective antibiotic-free solution for combating bacterial infections.

Engineering a transport pathway to boost extracellular production of human lactoferrin in Komagataella phaffii

Human lactoferrin (HLF), a pivotal iron-binding glycoprotein belonging to the transferrin family, is the primary immune protein in breast milk. Currently, recombinant HLF expression in microorganisms has attracted widespread attention. Through genome integration and copy number optimization, this study established a Komagataella phaffii cell factory that achieved an HLF titer of 137.6 mg/L. However, HLF accumulated extensively within the cells. Subsequently, several endogenous signal peptides were screened and hybridized with mating factor α, resulting in a 1.56-fold increase in extracellular HLF titers compared to the control. Additionally, protein kinase II overexpression facilitated vesicle trafficking, further increasing extracellular HLF to a titer of 304.6 mg/L in shake flasks and 1.7 g/L in a 3-L bioreactor. Notably, the intracellular/extracellular HLF concentration ratio decreased from 1.69:1 to 1.01:1. Further enhancements were made through ion optimization and an exponential methanol-feeding strategy, resulting in an extracellular HLF titer of 3.1 g/L. To the best of our knowledge, this is the first study to report considerably improved extracellular HLF production through optimization of the transport and secretion pathways and provide insights into the expression of other secretary human proteins in K. phaffii.

Hemochromatosis: Ferroptosis, ROS, Gut Microbiome, and Clinical Challenges with Alcohol as Confounding Variable.

Hemochromatosis represents clinically one of the most important genetic storage diseases of the liver caused by iron overload, which is to be differentiated from hepatic iron overload due to excessive iron release from erythrocytes in patients with genetic hemolytic disorders. This disorder is under recent mechanistic discussion regarding ferroptosis, reactive oxygen species (ROS), the gut microbiome, and alcohol abuse as a risk factor, which are all topics of this review article. Triggered by released intracellular free iron from ferritin via the autophagic process of ferritinophagy, ferroptosis is involved in hemochromatosis as a specific form of iron-dependent regulated cell death. This develops in the course of mitochondrial injury associated with additional iron accumulation, followed by excessive production of ROS and lipid peroxidation. A low fecal iron content during therapeutic iron depletion reduces colonic inflammation and oxidative stress. In clinical terms, iron is an essential trace element required for human health. Humans cannot synthesize iron and must take it up from iron-containing foods and beverages. Under physiological conditions, healthy individuals allow for iron homeostasis by restricting the extent of intestinal iron depending on realistic demand, avoiding uptake of iron in excess. For this condition, the human body has no chance to adequately compensate through removal. In patients with hemochromatosis, the molecular finetuning of intestinal iron uptake is set off due to mutations in the high-FE2+ (HFE) genes that lead to a lack of hepcidin or resistance on the part of ferroportin to hepcidin binding. This is the major mechanism for the increased iron stores in the body. Hepcidin is a liver-derived peptide, which impairs the release of iron from enterocytes and macrophages by interacting with ferroportin. As a result, iron accumulates in various organs including the liver, which is severely injured and causes the clinically important hemochromatosis. This diagnosis is difficult to establish due to uncharacteristic features. Among these are asthenia, joint pain, arthritis, chondrocalcinosis, diabetes mellitus, hypopituitarism, hypogonadotropic hypogonadism, and cardiopathy. Diagnosis is initially suspected by increased serum levels of ferritin, a non-specific parameter also elevated in inflammatory diseases that must be excluded to be on the safer diagnostic side. Diagnosis is facilitated if ferritin is combined with elevated fasting transferrin saturation, genetic testing, and family screening. Various diagnostic attempts were published as algorithms. However, none of these were based on evidence or quantitative results derived from scored key features as opposed to other known complex diseases. Among these are autoimmune hepatitis (AIH) or drug-induced liver injury (DILI). For both diseases, the scored diagnostic algorithms are used in line with artificial intelligence (AI) principles to ascertain the diagnosis. The first-line therapy of hemochromatosis involves regular and life-long phlebotomy to remove iron from the blood, which improves the prognosis and may prevent the development of end-stage liver disease such as cirrhosis and hepatocellular carcinoma. Liver transplantation is rarely performed, confined to acute liver failure. In conclusion, ferroptosis, ROS, the gut microbiome, and concomitant alcohol abuse play a major contributing role in the development and clinical course of genetic hemochromatosis, which requires early diagnosis and therapy initiation through phlebotomy as a first-line treatment.

The effectiveness of oral bovine lactoferrin compared to iron supplementation in patients with a low hemoglobin profile: A systematic review and meta-analysis of randomized clinical trials

BackgroundPatients with a low serum blood hemoglobin concentration suffer from a pathologic state that contributes significantly to morbidity and mortality figures worldwide. Oral iron supplementation, the most common method of treatment, is reported to have poor patient adherence, due to its unwanted side effects. Lactoferrin is a globular glycoprotein of the transferrin family that has shown promising results in patients with a low hemoglobin profile. This systematic review and meta-analysis of randomized clinical trials explore its effect on blood hemoglobin compared to conventional iron preparations.MethodsWe followed the PRISMA Guidelines for reporting systematic reviews and meta-analyses. A systematic search was conducted in electronic databases (PubMed, CINAHL, Scopus, and Cochrane) from inception to June 2022. Meta-analysis was performed on studies where the primary outcome was the mean Hb concentration, comparing lactoferrin to ferrous sulfate subgroups. We assessed the methodological quality of the trials using the Jadad scoring scale.ResultsNineteen trials published between 2006 and 2022 met the eligibility criteria. It has been found that the levels of Hb concentration in different populations with varying health conditions undergo a moderate to significant change after treatment with all types of trialed interventions, including both iron and lactoferrin treatment, in both the intervention group and the comparison group. Most of the studies report that LF showed a statistically significant increase in Hb concentration levels, compared to those in the iron group. The meta-analysis included seven trials comparing the effectiveness of lactoferrin to ferrous sulfate for patients with low Hb concentration. The analysis showed a statistically significant increase in Hb levels in the oral bovine lactoferrin group compared to ferrous sulfate (SMD -0.81, 95% CI: -1.21, -0.42, p < 0.0001, I2 = 95.8%, P heterogeneity < 0.001).ConclusionsLactoferrin is an effective intervention at doses of 100–250 ng/day, for patients with a low Hb concentration. As a safer option and with high compliance evidence, lactoferrin can serve as an iron replacement treatment for patients who may be experiencing adverse side effects due to iron intake.

Quantum chemical calculations on calcium oxalate and dolichin A and their binding efficacy to lactoferrin: An &lt;i&gt;in silico&lt;/i&gt; study using DFT, molecular docking, and molecular dynamics simulations

&lt;abstract&gt; &lt;p&gt;Lactoferrin, a member of the transferrin family, is one of the promoter proteins for calcium oxalate-type kidney stone formation. It exhibits a remarkable ability to interact with metals and oxalate ions. The prevalence of calcium oxalate in kidney stones was confirmed by the Fourier transform infrared spectra. The quantum chemical properties of calcium oxalate and dolichin A calculated by density functional theory and time-dependent density functional theory indicate their potential for hydrogen bonding and nonbonding interactions with the receptor proteins. From molecular docking analysis, the binding free energy of dolichin A was −7.78 kcal/mol, which was the best of twenty-four phytochemicals from &lt;italic&gt;Macrotyloma uniflorum&lt;/italic&gt;, and that of calcium oxalate was −3.86 kcal/mol to lactoferrin. Furthermore, dolichin A having favorable physicochemical and pharmacokinetic properties offers post molecular dynamics molecular mechanics generalized Born surface area free energy of −17.61 ± 4.03 kcal/mol, indicating the strong binding interactions, and, therefore, it acts as a potential inhibitor of the lactoferrin.&lt;/p&gt; &lt;/abstract&gt;

The Multifaceted Roles of Bovine Lactoferrin: Molecular Structure, Isolation Methods, Analytical Characteristics, and Biological Properties.

Bovine lactoferrin (bLF) is widely known as an iron-binding glycoprotein from the transferrin family. The bLF molecule exhibits a broad spectrum of biological activity, including iron delivery, antimicrobial, antiviral, immunomodulatory, antioxidant, antitumor, and prebiotic functions, thereby making it one of the most valuable representatives for biomedical applications. Remarkably, LF functionality might completely differ in dependence on the iron saturation state and glycosylation patterns. Recently, a violently growing demand for bLF production has been observed, mostly for infant formulas, dietary supplements, and functional food formulations. Unfortunately, one of the reasons that inhibit the development of the bLF market and widespread protein implementation is related to its negligible amount in both major sources─colostrum and mature milk. This study provides a comprehensive overview of the significance of bLF research by delineating the key structural characteristics of the protein and elucidating their impact on its physicochemical and biological properties. Progress in the development of optimal isolation techniques for bLF is critically assessed, alongside the challenges that arise during its production. Furthermore, this paper presents a curated list of the most relevant instrumental techniques for the characterization of bLF. Lastly, it discusses the prospective applications and future directions for bLF-based formulations, highlighting their potential in various fields.

Perspective: A proposal on solutions of modern supply chain construction for lactoferrin

Lactoferrin is an iron-binding glycoprotein of the transferrin family that is found in most bodily fluids of mammals and has a variety of biological and beneficial functions, with great importance in health enhancement as a supplement for humans and other animals. More than 300 t of lactoferrin were produced in 2021, and this number is expected to grow yearly by 10% to 12%, to over 580 t in 2030. With new and important functions of lactoferrin being revealed and studied, focus on its industrial production and application is increasing accordingly. However, lactoferrin is mainly sourced from cheese whey or skim milk by cation-exchange column chromatography, which is a costly and low-quality method. A potential solution for lactoferrin global supply chain construction is proposed in this article as a complement to traditional routes of purification from whey or skim milk. The large-scale production of lactoferrin, mainly by recombinant yeast, mammal, and grain systems, as well as the market niche and product design, are discussed.

Recombinant Human Lactoferrin Augments Epirubicin Chemotherapy in Solid Ehrlich Carcinoma Bearing Mice.

Lactoferrin (LF) is a member of the transferrin family, which is known for its immunomodulatory properties. LF has been widely used as an anticancer medication in various cancers including breast cancer. The current study aimed to examine the molecular mechanisms underlying the therapeutic potential of recombinant human lactoferrin (rhLF), either alone or combined with epirubicin (EPI), in mice bearing solid Ehrlich carcinoma (SEC). SEC-bearing female mice (n=40) were divided into 4 equal groups. Mice were given rhLF orally (100mg/kg/mouse) daily and/or EPI i.p (8mg/kg/mouse). The experiment lasted 14 days, after which samples were collected to measure IL-18 and phosphorylated c-Jun N-terminal kinase (p-JNK) by ELISA and p53 gene expression by real-time PCR. Administration of rhLF, either alone or combined with EPI, markedly decreased the tumor volume and increased tumor inhibition rate as well as survival rate compared to either tumor control group or EPI-mono treated group. In addition, co-administration of rhLF and EPI increased the level of activated JNKs and expression of p53 in tumor tissues compared to the tumor, control group, exhibiting their pro-apoptotic properties. Moreover, the combined treatment with rhLF and EPI elevated IL-18 level in the intestinal mucosa compared to other experimental groups with a possible immune-enhancing effect. Recombinant human lactoferrin exhibited potential anticancer and immune-enhancing properties in mice with breast cancer. Co-treatment with rhLF and EPI proved to be a promising strategy in cancer treatment.

Lactoferrin Modulates Induction of Transcription Factor c-Fos in Neuronal Cultures

Lactoferrin (Lf) is a multifunctional protein from the transferrin family. Of particular interest is the ability of Lf to affect a wide range of neuronal processes by modulating the expression of genes involved in long-term neuroplasticity. The expression of the immediate early gene c-fos that is rapidly activated in response to external influences, and its product, transcription factor c-Fos, is widely used as a marker of long-term neuronal plasticity. The present study aims to examine the effect of human Lf on the induction of transcription factor c-Fos in the primary mouse neuronal cultures after stimulation and to determine the cellular localization of human Lf and its colocalization with induced c-Fos protein. Primary dissociated cultures of hippocampal cells were obtained from the brains of newborn C57BL/6 mice (P0-P1). On day 7 of culturing, human Lf was added to the medium. After 24 h (day 8 in culture), c-Fos protein was induced in cells by triple application of 50 mM KCl. c-Fos content was analyzed using the immunofluorescent method 2 h after stimulation. Stimulation promoted exogenous Lf translocation into the nuclei of cultured neuronal cells, which correlated with increased induction of transcription factor c-Fos and was accompanied by nuclear colocalization of these proteins. These results attest to the potential of Lf as a modulator of neuronal processes and open up new prospects in studying the mechanisms of the regulatory effects of lactoferrin on cell function.

Lactoferrin and Nanotechnology: The Potential for Cancer Treatment.

Lactoferrin (Lf)-a glycoprotein of the transferrin family-has been investigated as a promising molecule with diverse applications, including infection inhibition, anti-inflammation, antioxidant properties and immune modulation. Along with that, Lf was found to inhibit the growth of cancerous tumors. Owing to unique properties such as iron-binding and positive charge, Lf could interrupt the cancer cell membrane or influence the apoptosis pathway. In addition, being a common mammalian excretion, Lf offers is promising in terms of targeting delivery or the diagnosis of cancer. Recently, nanotechnology significantly enhanced the therapeutic index of natural glycoproteins such as Lf. Therefore, in the context of this review, the understanding of Lf is summarized and followed by different strategies of nano-preparation, including inorganic nanoparticles, lipid-based nanoparticles and polymer-based nanoparticles in cancer management. At the end of the study, the potential future applications are discussed to pave the way for translating Lf into actual usage.

Antimicrobial Effects of Lactoferrin against Helicobacter pylori Infection.

Helicobacter (H.) pylori is the primary causative agent of various gastroduodenal diseases. H. pylori is an adapted microorganism that has evolved to survive in the acidic conditions of the human stomach, possessing a natural strategy for colonizing harsh environments. Despite the implementation of various eradication regimens worldwide, the eradication rate of H. pylori has decreased to less than 80% in recent years due to the emergence of antibiotic-resistant strains. This has posed a significant challenge in treating H. pylori infection, as antibiotic resistance and side effects have become increasingly problematic. Lactoferrin, a member of the transferrin family, is an iron-binding protein with antioxidant, antibacterial, antiviral, and anti-inflammatory properties that promote human health. The concentrations of lactoferrin in the gastric juice and mucosa significantly increase during H. pylori infection and are strongly correlated with the severity of gastric mucosal inflammation. Numerous researchers have studied the antimicrobial properties of lactoferrin both in vitro and in vivo. In addition, recent studies have investigated the addition of oral lactoferrin supplementation to H. pylori eradication therapy, even though monotherapy with lactoferrin does not eradicate the microorganism. In this article, we reviewed the survival strategy of H. pylori to evade the antimicrobial activity of human lactoferrin and explore the potential of lactoferrin in H. pylori eradication therapy.

Nutraceutical and Health-Promoting Potential of Lactoferrin, an Iron-Binding Protein in Human and Animal: Current Knowledge.

Lactoferrin is a natural cationic iron-binding glycoprotein of the transferrin family found in bovine milk and other exocrine secretions, including lacrimal fluid, saliva, and bile. Lactoferrin has been investigated for its numerous powerful influences, including anticancer, anti-inflammatory, anti-oxidant, anti-osteoporotic, antifungal, antibacterial, antiviral, immunomodulatory, hepatoprotective, and other beneficial health effects. Lactoferrin demonstrated several nutraceutical and pharmaceutical potentials and have a significant impact on improving the health of humans and animals. Lactoferrin plays a critical role in keeping the normal physiological homeostasis associated with the development of pathological disorders. The current review highlights the medicinal value, nutraceutical role, therapeutic application, and outstanding favorable health sides of lactoferrin, which would benefit from more exploration of this glycoprotein for the design of effective medicines, drugs, and pharmaceuticals for safeguarding different health issues in animals and humans.

Apo-form of recombinant human lactoferrin changes the genome-wide DNA methylation level and the chromatin compaction degree in neuroblastoma cell line IMR-32

BACKGROUND: Neuroblastoma is one of the most common extracranial solid tumors in childhood. At present, epigenetic disorders play a significant role in neoplasms development. Since epigenetic changes in the cell are quite dynamic and reversible, epigenome-modulating exogenous agents can be used in epigenetic targeted therapy for various types of tumors. Therefore, the identification of these agents is still significant. Lactoferrin is one such potential molecule from the transferrin family. Currently, the anti-tumor properties of lactoferrin have been identified, but its effect on the epigenome of cells of various tumors types, particularly on neuroblastomas, is practically unknown.&#x0D; AIM: To study the effect of the exogenous recombinant human apolactoferrin on the viability and epigenomic status of IMR-32 neuroblastoma cells.&#x0D; MATERIALS AND METHODS: We studied human IMR-32 neuroblastoma cells after 72 hours of exposure to 8 doses of recombinant human apolactoferrin: 0.1, 0.5, 1, 5, 10, 50, 100 and 500 g/ml. The level of genome-wide DNA methylation and the degree of chromatin compaction in IMR-32 cells were quantified using commercial kits 5-mC DNA ELISA Kit, Global DNA Methylation LINE-1 Kit, as well as enzymatic hydrolysis of MspI / HpaII and DNaseI.&#x0D; RESULTS: The recombinant apolactoferrin reduces the viability of IMR-32 and, depending on the dose, differentially affects the level of genome-wide DNA methylation (СpG dinucleotides, CCGG sites, LINE-1 repeats) and the degree of chromatin compaction. At the same time, a complex picture of the epigenomic cellular response to the effect of apo-lactoferrin was observed (nonlinear nonmonotonic dose-effect relationship).&#x0D; CONCLUSIONS: We assumed that apolactoferrin modulates gene activity through epigenetic mechanisms, in particular, by changing the DNA methylation pattern and affecting the chromatin structure, which may be one of the molecular mechanisms of its anti-tumor effect.

Human lactoferrin modulates gene expression of the cytokine IL4 and the receptor TLR4 in the rat spleen under stress and upon the lipopolysaccharide administration

ABSTRACT Lactoferrin is a multifunctional glycoprotein of the transferrin family with a molecular mass of about 80 kDa. We studied the effect of human lactoferrin on stress- and lipopolysaccharide-induced changes in blood corticosterone levels, as well as on the gene expression of the cytokine IL4 and the pattern-recognition receptor TLR4 in rat splenocytes. Stress in rats was modeled by swimming in cold water (1-4°C) for 2 min. Lactoferrin and lipopolysaccharide (LPS) were administered intraperitoneally before the stress exposure. Corticosterone level in the plasma was determined by enzyme immunoassay, and changes in gene expression were assessed by real-time polymerase chain reaction (PCR) with reverse transcription. We showed that preventive intraperitoneal administration of lactoferrin reduced the stress and LPS-induced increase in the gene expression of both IL4 and TLR4 in rat splenocytes but did not change the concentration of corticosterone in the blood.

Rapid and sensitive determination of lactoferrin in milk powder by boronate affinity amplified dynamic light scattering immunosensor

Lactoferrin (Lf) is a bioactive multifunctional glycoprotein that belongs to the transferrin family. As an important nutritional fortifier, Lf is one of the star nutrients in milk powder. Thus, the rapid and accurate quantitative detection of Lf content in milk powder is crucial to the evaluation of its quality. However, most reported immunoassay methods for Lf quantitation suffer from cumbersome sample pretreatment or lengthy testing time and do not meet the requirements for rapid Lf detection in milk powder. Herein, we report a novel method combining dynamic light scattering (DLS) immunosensing and boronate affinity recognition for the rapid and sensitive detection of Lf. Owing to the selective binding between a boronate ligand and the cis diol, the developed DLS immunosensor quantitatively detected Lf with saving one antibody, greatly reducing the need for matching antibodies, which are needed in traditional sandwich immunoassay for biomolecular detection. In addition, the DLS immunosensor exhibited high selectivity and sensitivity to Lf, with a detection limit of 1 ng/mL within ∼15 min through coupling with magnetic nanoparticle-assisted immunomagnetic separation and DLS signal transduction.

Lactoferrin: from the structure to the functional orchestration of iron homeostasis.

Iron is by far the most widespread and essential transition metal, possessing crucial biological functions for living systems. Despite chemical advantages, iron biology has forced organisms to face with some issues: ferric iron insolubility and ferrous-driven formation of toxic radicals. For these reasons, acquisition and transport of iron constitutes a formidable challenge for cells and organisms, which need to maintain adequate iron concentrations within a narrow range, allowing biological processes without triggering toxic effects. Higher organisms have evolved extracellular carrier proteins to acquire, transport and manage iron. In recent years, a renewed interest in iron biology has highlighted the role of iron-proteins dysregulation in the onset and/or exacerbation of different pathological conditions. However, to date, no resolutive therapy for iron disorders has been found. In this review, we outline the efficacy of Lactoferrin, a member of the transferrin family mainly secreted by exocrine glands and neutrophils, as a new emerging orchestrator of iron metabolism and homeostasis, able to counteract iron disorders associated to different pathologies, including iron deficiency and anemia of inflammation in blood, Parkinson and Alzheimer diseases in the brain and cystic fibrosis in the lung.