Lactoferrin Concentrations Research Articles

Interleukin-6 is associated with tryptophan metabolism and signs of depression in individuals with carbohydrate malabsorption.

The aim of the present study was to investigate possible associations between interleukin-6 (IL-6), interferon-gamma (INF-γ), tumor necrosis factor-alpha (TNF-α), lactoferrin and lipopolysaccharide binding protein (LBP) with TRP metabolism and signs of depression in a large cohort of outpatients referred for carbohydrate malabsorption testing. Serum concentrations of IL-6, INF-γ, TNF-α, lactoferrin, LBP, tryptophan (TRP), kynurenine (KYN) and kynuric acid were determined in 250 adults referred for lactose and fructose malabsorption testing. All participants filled out the Beck Depression Inventory (BDI). Serum IL-6 levels were positively correlated with the BDI score (p = 0.001, ρ = 0.205) and indicators of TRP metabolism (KYN/TRP ratio, KYN) (P-values < 0.05, ρ = 0.176 and 0.136). Ninety-five individuals with a BDI score > 13 showed significantly higher IL-6 serum levels (1.7 [1.0 - 2.8] vs. 1.1 [0.8 - 1.7] pg/mL, p < 0.001) compared to 115 individuals with a BDI score ≤ 13. LBP showed a positive correlation with the KYN/TRP ratio (p = 0.005, ρ = 0.177). IL-6 and LBP were associated with indicators of TRP metabolism. IL-6 was found to be linked to signs of depression. Individuals with the presence of depressive symptoms showed higher serum IL-6 levels compared to individuals without depressive symptoms.

Plasma lactoferrin levels in newborn infants with early-onset neonatal sepsis

Objective: To determinate if lactoferrin (LF) could be optimal biomarker; the LF levels in plasma were compared between neonates with or without early-onset neonatal sepsis (EOS). Methods: From January to December of 2019 we conducted an analytic cross-sectional study, eighty-nine patients were collected: 34 newborns with EOS and 55 newborns without EOS from neonatology unit from a tertiary care hospital. The diagnosis was made with clinical parameters and sepsis biomarkers. Plasma concentrations of LF were measured by Enzyme-Linked Immunosorbent Assay (ELISA). Results: The LF median was 3.7ng/ml to the newborn group with EOS while 44.8ng/ml was to newborn without EOS (p: 0.000). The best LF cut-off point in newborns with EOS was 20.55ng/ml, with sensitivity of 73.5%, specificity 92.7%, positive predictive value 86.2%, and negative predictive value 85%, positive likelihood ratio 10.11, and negative likelihood ratio 0.28. Conclusions: These findings indicate that levels of LF in plasma could be an effective and useful biomarker to diagnose of EOS.

A comparison of 4 different machine learning algorithms to predict lactoferrin content in bovine milk from mid-infrared spectra

Lactoferrin (LF) is a glycoprotein naturally present in milk. Its content varies throughout lactation, but also with mastitis; therefore it is a potential additional indicator of udder health beyond somatic cell count. Condequently, there is an interest in quantifying this biomolecule routinely. First prediction equations proposed in the literature to predict the content in milk using milk mid-infrared spectrometry were built using partial least square regression (PLSR) due to the limited size of the data set. Thanks to a large data set, the current study aimed to test 4 different machine learning algorithms using a large data set comprising 6,619 records collected across different herds, breeds, and countries. The first algorithm was a PLSR, as used in past investigations. The second and third algorithms used partial least square (PLS) factors combined with a linear and polynomial support vector regression (PLS + SVR). The fourth algorithm also used PLS factors, but included in an artificial neural network with 1 hidden layer (PLS + ANN). The training and validation sets comprised 5,541 and 836 records, respectively. Even if the calibration prediction performances were the best for PLS + polynomial SVR, their validation prediction performances were the worst. The 3 other algorithms had similar validation performances. Indeed, the validation root mean squared error (RMSE) ranged between 162.17 and 166.75 mg/L of milk. However, the lower standard deviation of cross-validation RMSE and the better normality of the residual distribution observed for PLS + ANN suggest that this modeling was more suitable to predict the LF content in milk from milk mid-infrared spectra (R2v = 0.60 and validation RMSE = 162.17 mg/L of milk). This PLS +ANN model was then applied to almost 6 million spectral records. The predicted LF showed the expected relationships with milk yield, somatic cell score, somatic cell count, and stage of lactation. The model tended to underestimate high LF values (higher than 600 mg/L of milk). However, if the prediction threshold was set to 500 mg/L, 82% of samples from the validation having a content of LF higher than 600 mg/L were detected. Future research should aim to increase the number of those extremely high LF records in the calibration set.

Translocation of intrauterine-infused bacterial lipopolysaccharides to the mammary gland in dexamethasone-treated goats.

Our previous study showed that intrauterine-infused lipopolysaccharide (LPS) can be translocated to the mammary gland to induce weak inflammation. This study aimed to determine whether dexamethasone treatment facilitated the translocation of LPS from the uterus to the mammary gland to induce a heavy inflammatory response. Sixteen goats were divided into control and LPS groups, subjected to daily dexamethasone administration before saline or LPS infusion. Milk and blood samples were collected before and after LPS infusion to determine the milk yield and somatic cell count (SCC) and blood leucocyte count (BLC), cytokines, antimicrobial peptides and serum amyloid A (SAA) concentrations. Mammary gland tissues were collected from two goats before and 24hr after LPS infusion for immunohistochemical analysis of LPS. The mean SCC in the LPS group was significantly higher, whereas the milk yield was significantly lower than that in the control group after LPS infusion. The mean BLC in the LPS group was significantly lower than in the control group after LPS infusion. Furthermore, milk concentrations of IL-1β, S100A8 and lactoferrin were higher in the LPS group than in the control group after infusion. LPS was detected in the connective tissues and inner alveolar spaces of the mammary glands 24hr after LPS infusion. We concluded that dexamethasone administration facilitated the translocation of intrauterine-infused LPS to the mammary gland, where it induced an inflammatory response. Therefore, LPS translocated from other organs, such as the uterus, can induce heavy inflammation in the mammary gland under immunosuppressive conditions.

Lactoferrin Concentration in Human Tears and Ocular Diseases: A Meta-Analysis.

PurposeTo evaluate the potential of lactoferrin (Lf) as a diagnostic biomarker for ocular diseases using a meta-analytic approach.MethodsAll original studies reporting an estimate of the average Lf concentration in healthy subjects and those affected by ocular diseases were searched up to March 2020. The DerSimonian and Laird method was used to calculate the random effects pooled mean difference and the corresponding 95% confidence interval (CI) in Lf concentration between healthy subjects and those affected by dry eye (DE), Sjögren syndrome (SS), and diabetic retinopathy, separately. The presence of between-study heterogeneity was evaluated using the Cochran's Q test and the I2 index. Stratified analyses were performed to assess potential sources of heterogeneity and influence and cumulative analyses to evaluate the robustness of the results obtained. Publication bias was also evaluated using funnel plot and the Egger's test.ResultsThe pooled mean differences in Lf concentrations between healthy subjects and those with DE, Sjögren syndrome, and diabetic retinopathy were respectively 0.62 (95% CI, 0.35–0.89) for DE, 3.78 (95% CI, −6.64 to 14.17), and 0.19 (95% CI, −4.00 to 4.39). Regarding DE, the stratified analysis showed that geographical area (P value Q test < 0.0001) and sample size (P < 0.0005) were sources of heterogeneity. Moreover, no study substantially influenced the results obtained and the pooled mean difference became statistically significant after a sample size of 220. Publication bias may affect the results of DE.ConclusionsThe results of the current meta-analysis suggest that Lf level in tears is a good candidate as dry eye syndrome diagnostic biomarker.

The effect of freeze-drying and storage on lysozyme activity, lactoferrin content, superoxide dismutase activity, total antioxidant capacity and fatty acid profile of freeze-dried human milk

Pooled human milk samples were freeze-dried and stored for 6 weeks at a temperature of 5 °C and 25 °C. Freeze-drying decreased the water content of milk by 86.5%, and the obtained lyophilizate was readily soluble in water. The freeze-drying process did not affect superoxide dismutase (SOD) activity, fatty acid (FA) profile or lactoferrin (LF) content, but it decreased total antioxidant capacity (TAC) of human milk by 22.1% and induced a minor increase in lysozyme (LZ) activity, by approximately 9.8%. Storage of freeze-dried milk did not show significant influence on TAC, LF, FA and LZ levels, while after six weeks of storage SOD activity decreased by around 27% relative to the level noted immediately after lyophilization. These findings and the remaining state of knowledge imply, that freeze-drying can be a useful method of human milk storage. Highlights Lyophilization does not significantly affect the level of SOD and LF of human milk. In freeze-dried milk, bioactive substances: TAC level, LF, FA and LZ, remain stable even at room temperature. During storage of freeze-dried milk SOD activity decreases significantly.

Effects of exercise in hot and humid conditions and bovine colostrum on salivary immune markers.

The purpose of this study was to examine the effects of exercise in a hot and humid environment on salivary lactoferrin and lysozyme. A secondary aim was to quantify the effects of 14-day bovine colostrum (BC) supplementation on salivary lactoferrin and lysozyme at rest and following exercise in hot and humid conditions. Using a randomized, double-blind, and counterbalanced design, ten males (20±2 years, VO2max 55.8±3.7mLkg-1min-1, 11.8±2.7% body fat) ran for 46±7.7minat 95% of ventilatory threshold in a 40°C and 50% RH environment after 14-days of supplementation with either BC or placebo. Saliva was collected pre, post, 1-h, and 4-h post exercise, and was analyzed for lactoferrin and lysozyme using ELISA. There was an immediate increase in the concentration and secretion rate of lactoferrin and lysozyme (p<0.05) with exercise, but BC had no effect (p>0.05). Saliva flow rate was not different between conditions [(PLA: pre: 0.54±0.3, post: 0.44±0.3, 1-h: 0.67±0.3, 4-h: 1.0±0.4mLmin-1); (BC: pre: 0.58±0.2, post: 0.37±0.1, 1-h: 0.63±0.2, 4-h: 0.83±0.4mLmin-1)]. There were no differences in thermoregulatory markers (core temperature or physiological strain index) between BC and placebo trials. Interestingly, exercise-induced heat stress did not impair mucosal immune parameters, instead participants showed a transient increase in salivary lactoferrin and lysozyme. Further, 14-day BC supplementation had no effect on mucosal immunity at any time point.

The Effect of Prolonged Freezing and Holder Pasteurization on the Macronutrient and Bioactive Protein Compositions of Human Milk.

Introduction: Human donor milk is widely used to feed premature and sick newborns when the milk of their own mothers is insufficient. All treatment processes involving human milk affect its composition. The aim of this study was to assess changes in the macronutrients and bioactive protein (lactoferrin and lysozyme) content in human milk caused by freezing and Holder pasteurization. Materials and Methods: Milk samples were collected from 42 mothers 14-16 days after delivery. Each sample was divided into two parts and tested twice for macronutrient content, once upon being freshly collected and again after freezing at -40°C, thawing and Holder pasteurization. The lysozyme and lactoferrin concentrations were first determined in the unpasteurized thawed human milk after it was stored frozen at -80°C for up to 10 months and again after Holder pasteurization. The macronutrient concentrations were determined by midinfrared spectrophotometry, and enzyme-linked immunosorbent assay was used to measure the lysozyme and lactoferrin concentrations. Results: Freezing and Holder pasteurization had no significant effects on the macronutrient concentrations. The mean lactoferrin content before and after pasteurization was 2.5 ± 1.07 and 0.03 ± 0.03 mg/mL, respectively (p < 0.001), and the lysozyme content was 19.57 ± 20.11 and 12.62 ± 14.14 μg/mL, respectively (p = 0.007). Conclusions: Freezing and Holder pasteurization did not decrease the nutritional value of human milk but caused considerable loss of lactoferrin and lysozyme. New methods for treating human milk are needed that ensure the destruction of pathogenic microorganisms while retaining the biological and nutritional value of the milk. The Clinical Trial Registration number: NCT04382989.

Sensitive immunoassays based on specific monoclonal IgG for determination of bovine lactoferrin in cow milk samples

Lactoferrin (LF), a bioactive multifunctional protein of the transferrin family, is found mainly in the secretions of all mammals, especially in milk. In the present study, a hybridoma cell (LF8) secreting IgG against bovine LF was screened, and the purified LF8 mAb showed high specificity and affinity to bovine LF. The linear range of ic-ELISA to detect LF was 9.76 ~ 625 ng/mL, with a limit of detection (LOD) of 0.01 ng/mL. The average recovery of intra- and inter-assay were (104.45 ± 4.12)% and (107.13 ± 4.72)%, respectively. The LOD of colloidal gold- and AuNFs-based strip by naked eye were 9.7 and 2.4 ng/mL, respectively, and the detection time was less than 10 min without any samples pretreatment and expensive equipment. The developed ELISA and lateral flow immunosensors based on specific IgG could be used directly for rapid detection of the bovine LF content in cow milk samples.

Lactoferrin is a dynamic protein in human melioidosis and is a TLR4-dependent driver of TNF-α release in Burkholderia thailandensis infection in vitro.

Melioidosis is an often-severe tropical infection caused by Burkholderia pseudomallei (Bp) with high associated morbidity and mortality. Burkholderia thailandensis (Bt) is a closely related surrogate that does not require BSL-3 conditions for study. Lactoferrin is an iron-binding glycoprotein that can modulate the innate inflammatory response. Here we investigated the impact of lactoferrin on the host immune response in melioidosis. Lactoferrin concentrations were measured in plasma from patients with melioidosis and following ex vivo stimulation of blood from healthy individuals. Bt growth was quantified in liquid media in the presence of purified and recombinant human lactoferrin. Differentiated THP-1 cells and human blood monocytes were infected with Bt in the presence of purified and recombinant human lactoferrin, and bacterial intracellular replication and cytokine responses (tumor necrosis factor-α (TNF-α), interleukin-1β and interferon-γ) were measured. In a cohort of 49 melioidosis patients, non-survivors to 28 days had significantly higher plasma lactoferrin concentrations compared to survivors (median (interquartile range (IQR)): 326 ng/ml (230–748) vs 144 ng/ml (99–277), p<0.001). In blood stimulated with heat-killed Bp, plasma lactoferrin concentration significantly increased compared to unstimulated blood (median (IQR): 424 ng/ml (349–479) vs 130 ng/ml (91–214), respectively; p<0.001). Neither purified nor recombinant human lactoferrin impaired growth of Bt in media. Lactoferrin significantly increased TNF-α production by differentiated THP-1 cells and blood monocytes after Bt infection. This phenotype was largely abrogated when Toll-like receptor 4 (TLR4) was blocked with a monoclonal antibody. In sum, lactoferrin is produced by blood cells after exposure to Bp and lactoferrin concentrations are higher in 28-day survivors in melioidosis. Lactoferrin induces proinflammatory cytokine production after Bt infection that may be TLR4 dependent.

Comparative effect of immunomodulators on the contents of hydrolase inhibitors and lactoferrin in community-acquired pneumonia in adults

The purpose of our study was to examine the effect of immunomodulators (broncho-vaxom, immunovac-VP4 vaccine and polyoxidonium) upon the kinetics of serum hydrolase inhibitors and lactoferrin in the treatment of community-acquired pneumonia (CAP). The study included 71 CAP patients at the age of 18 to 70 years. The patients were divided into 4 groups: Group I (15 people) was a control group treated with basic antibacterial and symptomatic therapy, according to the standard treatment regimen, without use of immunomodulators; the patients from group II (19 patients) were additionally administered bronchovaxom (the drug was prescribed upon admission: 1 course over 30 days, then 2 rounds for 10 days each, with an interval in 20 days); group III (20 cases) contained the patients who additionally received polyoxidonium (the drug was prescribed from the 1st day of hospitalization, 6 mg daily i/m for 3 days, then 10 injections over 10 days); group IV (17 cases): Immunovac-VP4 vaccine was administered orally 4 ml and intranasally 2 drops on days 1, 4, 7, 10, 13, 19, 25, 31, along with antibacterial and symptomatic therapy. This vaccine consists of antigens from opportunistic microorganisms (a multicomponent mixture of water-soluble antigens of S. aureus, K. pneumoniae, P. vulgaris, E. coli). Serum concentrations of α2-macroglobulin and α1-antitrypsin hydrolase inhibitors were determined by the method of quantitative immunoelectrophoresis using the research test systems; lactoferrin (LF) levels were evaluated by enzyme-linked immunosorbent assay using commercial test systems. These indicators were studied in blood serum before treatment, on the 2nd, 13th, and 60th days of observation. It was shown that administration of immune modulators combined with antibiotic therapy in patients with CAP can affect the kinetics of acute phase inflammatory proteins. The effect of broncho-vaxom corresponds to the classical pathway of the response to inflammatory process, i.e., activation of complex of positive acute phase reactants (α1-antitrypsin and lactoferrin) and inhibition (blocking) of negative acute phase reactants (α2macroglobulin). Polyoxidonium has a noticeable effect only upon neutrophils secreting lactoferrin. ImmunovacVP4 promotes only short-term secretion of this protein. One may assume that activation of hydrolase inhibitors and lactoferrin after use of immunotropic drugs enhances clinical effect of therapy, with decreased severity and duration of symptoms, as well as lower exacerbation risk of chronic diseases and lesser volume of drug intake. Antibacterial therapy does not significantly affect the kinetics of acute phase inflammation reactants in patients with CAP. Administration of immunomodulators in combination with standard basic therapy may affects the inflammatory process to different degree, thus, in turn, leading to improved prognosis in this disease.

Detection of lactoferrin in bovine and goat milk by enzyme-linked immunosorbent assay

Lactoferrin (LF) has been proposed as a multifaceted functional ingredient of food. A competitive enzyme-linked immunosorbent assay (ELISA) was established to determine the LF in milk by using bovine LF antiserum and bovine LF-Biotin conjugate. The LF concentration in cow milk samples could be determined by this ELISA. However, the LF concentration in goat milk could not be measured using this method. Thus, the goat LF ELISA was established using goat LF-Biotin conjugate to measure the LF concentration in goat milk. Milk samples were collected for measurement of their LF concentrations. The mean LF level for the individual cow milk samples with the somatic cell count (SCC) below 1 × 105, 1 to 2.5 × 105 or 2.5 × 105 to 5 × 105 cells/mL was found to be about 176, 466 or 742 μg/mL, respectively. On the other hand, the LF level was about 200 μg/mL in goat bulk milks with the methylene blue reduction test (MBRT) time more than 5 hr. In conclusion, both LF ELISA methods developed in this study could be successfully used to determine the LF concentration in goat and cow milk.

Lactoferrin and lactoferricin B reduce adhesion and biofilm formation in the intestinal symbionts Bacteroides fragilis and Bacteroides thetaiotaomicron

Several factors affect the composition of species that inhabit our intestinal tract, including mode of delivery, genetics and nutrition. Antimicrobial peptides and proteins secreted in the gastrointestinal tract are powerful tools against bacteria. Lactoferrin (LF) inhibits the growth of several bacterial species, such as Enterobacteriaceae, but may stimulate probiotic bacteria. Activity of LF against gut symbiotic species of the Bacteroides genus could give us insights on how these species colonize the gut. We investigated the effects of the antimicrobial protein lactoferrin and its derived peptide, lactoferricin B on two species of strict anaerobes, opportunistic pathogens that cause diseases in both adults and children, commonly found in the microbiota of the human gastrointestinal tract, Bacteroides fragilis and B. thetaiotaomicron., In vitro biofilm formation and binding to laminin were strongly inhibited by a low concentration of lactoferrin (12.5 μg/ml). Conversely, the growth of the strains in a micro-dilution assay in minimal media with different iron sources was not affected by physiological concentrations (2 mg/ml) of apo-lactoferrin or holo-lactoferrin. The combination of lactoferrin with antibiotics in synergism assays was also negative. The lactoferricin B fragment was also unable to inhibit growth in a similar test with concentrations of up to 32 μg/ml. Resistance to lactoferrin could confer an advantage to these species, even when high amount of this protein is present in the gastrointestinal tract. However, colonization is hampered by the binding and biofilm inhibitiory effect of lactoferrin, which may explain the low prevalence of Bacteroides in healthy babies. Resistance to this antimicrobial protein may help understand the success of these opportunistic pathogens during infection in the peritoneum.

Purification of lactoferrin from sweet whey using ultrafiltration followed by expanded bed chromatography

Lactoferrin is an important protein found in sweet whey that has several biological activities, such as nutritional, antimicrobial, antiviral, anti-inflammatory, antioxidant, anticarcinogenic and immunomodulation. However, it is found in very low concentration in whey and its capture and purification is a challenge. This study proposes a simple strategy in which two operations for the separation and purification of lactoferrin are integrated. Sweet whey containing 0.12 mg/mL of lactoferrin was subjected to microfiltration for clarification, followed by ultrafiltration and diafiltration in order to reduce in 10 times the total volume with simultaneous concentration and pre-purification of lactoferrin to 1.1 mg/mL. In the second operation, the protein was purified by a preparative chromatography based on cationic exchange expanded bed chromatography (EBC). Adsorption isotherms were determined which allowed establishing the conditions for the chromatographic step. Hydrodynamic studies of the expanded bed and its dynamic binding capacity were also accomplished. The final concentration of the purified lactoferrin was 17.4 mg/mL, resulting in a purity of 92.7% and recovery of 87.0%.

Anticancer Effect of Bovine Lactoferrin on Breast Cancer Cell Line MCF7 and the Evaluation of Bax and Bak Genes Expression Involved in Apoptosis

Background: Lactoferrin is a glycoprotein with antimicrobial, antioxidant, immune-modulating, antiviral, and most importantly anticancer properties. In the present study, the effect of lactoferrin on breast cancer cell growth and the expression of Bax and Bak genes are evaluated. Materials and Methods: MCF7 cells were cultured in a 96-well plate with 1×105 cells in each well. Different lactoferrin concentrations of 0, 50, 300, 600, and 800 μg/mL were added to each well in three replicates and the well was incubated for 24 hours. After treatment, cell survival was measured using the MTT assay. To determine the level of expression of Bax and Bak genes, the cells were treated with lactoferrin concentrations of 0, 50, and 800 μg/mL in 2 replicates for 24 hours. Then RNA extraction was performed and cDNA was synthesized immediately and the expression of the genes in the presence of beta-actin reference gene and cyber-green fluorescence color was investigated with real-time reactions. Results: The cells viability in lactoferrin concentrations of 0, 50, 300, 500 and 800 μg/μL were 100%, 94%, 83%, 62%, and 32%, respectively. The expression level of the Bax gene at a concentration of 50 μg increased by 2.71 times and in 800 μg concentration decreased by 0.88 times. Also, the expression level of the Bak gene at concentrations of 50 and 800 μg increased by 1.23 and 1.0 fold, respectively. Statistical analysis of the data indicated that the expression levels of two genes at a concentration of 50 μg/mL of lactoferrin significantly increased (P&lt;0.01), compared to the control. The significance level in this study was set at &lt; 0.05. Conclusion: In this study, lactoferrin showed a growth inhibitory effect on breast cancer cells and increased the expression of Bax and Bak genes involved in apoptosis at a concentration of 50 µg/mL.

MODERN DIAGNOSTICS METHODS OF NON-SPECIFIC PROTECTION OF PERIODONTAL TISSUES IN RESIDENTS OF THE INDUSTRIAL REGION

The aim of the article is to study the peculiarities of antimicrobial immunity of the oral cavity of the industrial region’s inhabitants in order to optimize special preventive programs of major dental diseases.&#x0D; The object and research methods. 178 patients was examined from 21 to 50 years old, who turned to the University Dental Center of Zaporizhzhia State Medical University. The study group consisted of 126 patients with generalized periodontitis of the initial (8), I (32), II (68) and III (18) degree of chronic course, complicated by harmful factors of manufacturing. The comparison group consisted of 32 patients with periodontitis (5), I (10), II (11) and III (6) degrees who did not work in adverse conditions. The control group consisted of 20 relatively healthy individuals aged from 19 to 25 years without signs of generalized periodontitis.&#x0D; To study the factors of local immunity of the oral cavity, oral fluid was collected from each examined person, which was obtained without stimulation, spitting into sterile tubes. Then the oral fluid was centrifuged for 15 minutes at 8,000 rpm. The supernatant part of the oral liquid was poured into plastic tubes and stored at 30°C. The quantitative determination of markers in the oral fluid was performed by standard enzyme immunoassay kits according to the instructions of the "Lactoferrin-strip" ("VectorBest") manufacturers. The result was expressed in mµg / ml. The content of cathelicidin LL-37 was determined by enzyme-linked immunosorbent assay using NycultBiotechhuman LL-37 ELISA (Netherlands) firm reagent kit-manufacturers. The result was expressed in µg / ml. Statistica 13.0 licensed number JPZ804I382130ARCN10-J was used to process the results.&#x0D; The results of the study and their discussion. The hygienic condition in the examined patients of the study group was assessed as unsatisfactory: from 2.24 ± 0.5 to 2.99 ± 0.47 points depending on the age and length of service at the enterprise, but the PI and SBI values were increased.&#x0D; It was found, that the lactoferrin level in the oral fluid was higher in the all patients with periodontitis, than the same index in comparison with healthy control group and consistently increased with increasing severity of the process. In the patients’ study group, working in hazardous conditions of production, the lactoferrin content has increased in the oral fluid relative to the level of the healthy patients was more pronounced in comparison with the patients, suffering of periodontal tissue disease, but do not work in harmful manufacturing conditions. Increased lactoferrin in the oral fluid in the steelmaking workers can be considered as a means of compensation, that provides protection of the oral mucosa from colonization of microorganisms.&#x0D; In the steel industry workers, a statistically significant cathelicidin content decrease in the oral fluid was observed compared to healthy control patients’ group. In the clinical group of the patients without adverse factors of production, the cathelicidin’s concentration in the oral fluid was also reduced.&#x0D; The increasing level of lactoferrin in the oral fluid in parallel with the cathelicidin’s decrease in saliva are markers of the inflammatory phase, as well as the destructive phase of connective tissue.&#x0D; Conclusions. Thus, our observations have established a direct relationship between the lactoferrin’s concentration in the oral fluid and the severity of periodontal tissue diseases and the feedback between the cathelicidin’s content in the oral liquid and the activity of the pathological process.&#x0D; The determination of lactoferrin and cathelicidin in the oral fluid of the patients with harmful manufacturing factors allow us to identify the signs of pathological process in the oral cavity.

Regulation of the microecological state of the lower parts of the female reproductive tract with vitamins

Aim. To study the changes in the microecology of the vagina under the influence of ascorbic acid, α-tocopherol, and retinol acetate.Materials and methods. The number of vaginal lactobacilli and opportunistic bacteria, the concentrations of IL1β, raIL1, INFγ, TGFβ1, lactoferrin and sIgA in vaginal fluids of women with deficiency of ascorbic acid (n = 35) or retinol acetate (n = 35) or α-tocopherol (n = 35) were assessed before and after intravaginal application of these vitamins. The obtained data were compared with those of healthy women without vitamin deficiency (n = 15).Results. An association between deficiency of α-tocopherol, retinol acetate and ascorbic acid with a deficiency of lactoflora, a high frequency of bacterial vaginosis and candidiasis, and proinflammatory changes of the parameters of mucosal immunity has been found. A normalizing effect of all vitamins on the lactoflora of the vagina was identified. The severity of this effect decreased in the following order: α-tocopherol → retinol acetate → ascorbic acid. Retinol acetate caused the increase, while ascorbate and tocopherol caused the reduction in the number of opportunistic microorganisms. The effect of ascorbate was pronounced, but short-term, while the effect of tocopherol was less pronounced, but long-lasting. The use of vitamins affected the condition of mucosal immunity in different directions: ascorbic acid caused an increase in pro-inflammatory changes; on the contrary, α-tocopherol and retinol acetate caused anti-inflammatory changes.Conclusion. Our data on the effects of α-tocopherol, retinol acetate, and ascorbic acid on the microecological state of the vaginal biotope suggest that these vitamins may be considered as regulatory factors for both host cells and microsymbionts.

Lactoferrin levels in cervical fluid from in vitro fertilization (IVF) patients - correlation with IVF parameters.

Since our previous results suggest that lactoferrin (LF) might have roles in the reproductive process and that its levels might change in the female tract as a response to various factors, the aim of this investigation was to assess whether LF levels in cervical secretions correlate with reproductive parameters from invitro fertilization (IVF) patients. Cervical fluid samples were obtained from 34 women under 40years old enrolled for assisted reproduction techniques, and LF concentration was measured. The mean total protein concentration in all cervical fluid samples was 842.8± 116.9µg/mL. The mean concentration of LF was 0.73± 0.06ng LF/µg of total proteins. We observed that higher LF levels in cervical fluid correlated with lower IVF rates when all patients were analyzed; this negative correlation was also sustained when only patients ≥35 years were studied. The mean LF concentration in cervical fluid was significantly lower among patients with normal IVF rates than in those with values 50% or less. Using a LF cutoff value of 0.83ng/μg of total proteins, the study revealed a significant association between the LF levels below 0.83ng/µg of total proteins and IVF rates above 50%. LF levels in cervical mucus could potentially be used as a marker of fertilization outcome.

Differential impact of Holder and High Temperature Short Time pasteurization on the dynamic in vitro digestion of human milk in a preterm newborn model

The high-temperature short-time (HTST, 72 °C, 15 s) pasteurization of human milk (HM) has been proposed as an alternative to the Holder method (HoP, 62.5 °C, 30 min), to increase the preservation of bioactive compounds. We have investigated the impact of HTST and HoP pasteurization on the gastrointestinal kinetics of human milk, using a dynamic in vitro system in a preterm newborn model. An increased protein aggregation on the surface of fat globules following pasteurization, albeit to a lesser extent in HTST than in HoP, was observed. Despite relevant differences in the undigested milk samples, both pasteurization methods led to similar proteolytic patterns, while raw HM presented a higher native lactoferrin content throughout digestion. The slightly decreased amino acid release following HoP, with respect to HTST and raw HM, indicated that peptidomic analysis, which is currently underway, might provide interesting insights on the differential digestive kinetics of differently pasteurized HM.

Milk lactoferrin concentration of primiparous and multiparous sows during lactation

Lactoferrin (LF), a sialylated iron-binding glycoprotein, has numerous vital physiological functions including immunomodulation and protection against a large group of microorganisms, improving neurodevelopment, health, growth performance, and milk production. Lactoferrin occurs in human milk at a higher concentration compared with bovine milk, but little information is available on LF concentrations in porcine milk and the effects of sow parity on milk LF concentration. The objective of this study was to quantify the LF concentration in porcine milk and to compare that concentration between gilts and sows during lactation. We also investigated the effect of genetic background and litter size of the female pig on the LF concentration of porcine milk. The milk from 30 gilts and 35 sows was collected at 3 stages of lactation, namely colostrum, transition, and mature milk. Standard and experimental samples were analyzed by ultra-high performance liquid chromatography using a diode array UV detector. The following findings were reported: (1) porcine milk contained significant levels of LF with the highest concentration in colostrum, which decreased by ∼62% and ∼67% in transitional and mature milk, respectively; (2) mature gilt milk contained a 22% higher concentration of LF compared with sow milk, which was statistically significant; (3) breed line had an overall significant effect on the LF content of porcine milk; however, when the breed was considered, no significant difference was observed; and (4) LF concentration of porcine milk was not significantly influenced by the litter size. The presence of LF in a higher concentration in porcine milk suggests that LF is an important constituent of pig milk that might contribute to the optimum growth and development of piglets.