Developed and Validated Capillary Isotachophoresis Method for the Rapid Determining Organic Acids in Children's Saliva.

One of the current challenges facing researchers is the search for alternative biological material, as opposed to routinely and invasively collected (such as blood), as the analysis of the former would provide information about the state of human health, allowing for the diagnosis of diseases in their early stages. With the search for disease biomarkers in alternative materials, the development of newer analytical solutions has been observed. This study aims to develop a reliable analytical method using the capillary isotachophoresis technique for the determination of organic acids in children's saliva, the presence/elevation of which can be used in the future for diagnostic purposes. Organic acids such as formic, lactic, acetic, propionic, and butyric acid, were determined in the saliva of healthy children without carious lesions. The limit of quantification determined in the validation process was found to vary from 0.05 to 1.56 mg/L, the recoveries at the two levels were determined to vary between 90% and 110% for level I, while for level II the corresponding values of 75% and 106% were found; the presentation, expressed as relative standard deviation values (RSD), did not exceed 5%. The parameters determined while validating the results method indicated that the obtained are reliable. The Red-Green-Blue (RGB) additive color model was used for the evaluation of the method. This comparative analysis allowed us to define the color of the method, which expresses whether it meets the given assumptions and requirements. According to the RGB model, the isotachophoresis method developed requires less reagent input, shorter sample preparation times, and results with lower energy consumption. Thus, the subject procedure may provide an alternative, routine tool for determining organic acids in human saliva, to be applied in the diagnosing of diseases of various etiological origins.

The Importance of Nutritional Aspects in the Assessment of Inflammation and Intestinal Barrier in Patients with Inflammatory Bowel Disease.

Intestinal inflammation in inflammatory bowel disease (IBD) is closely linked to nutrition. This study aimed to evaluate associations between nutritional, inflammatory, and intestinal barrier parameters in patients with IBD. We assessed nutritional status, fecal short-chain fatty acid profile, serum cytokine levels, and mRNA expression of enzymes and tight junction proteins in intestinal biopsies obtained from 35 patients, including 11 patients with inactive IBD, 18 patients with active IBD, and six controls. Patients with active IBD were characterized by hypoalbuminemia, fluctuations in body weight, and restriction of fiber-containing foods. In addition, they had significantly reduced levels of isovaleric acid and tended to have lower levels of butyric, acetic, and propionic acids. Patients with active IBD had higher mRNA expression of peroxisome proliferator-activated receptor γ and inducible nitric oxide synthase, and lower mRNA expression of claudin-2 and zonula occludens-1, compared with patients with inactive IBD. Moreover, patients with a body mass index (BMI) of ≥25 kg/m2 had higher median tumor necrosis factor-α levels that those with a lower BMI. We comprehensively evaluated inflammatory parameters in relation to IBD activity and nutritional status. The discrepancies between proinflammatory and anti-inflammatory parameters depending on IBD activity may be related to nutritional factors, including diet and abnormal body weight.

Application of the Clustering Technique to Multiple Nutritional Factors Related to Inflammation and Disease Progression in Patients with Inflammatory Bowel Disease.

Diet and nutritional status affect intestinal inflammation in patients with inflammatory bowel disease (IBD). The aim of this study was to use a cluster analysis to assess structural similarity between different groups of parameters including short-chain fatty acid (SCFA) levels in stool as well as hematological and inflammatory parameters (such as serum C-reactive protein (CRP) and proinflammatory and anti-inflammatory cytokines). We also assessed similarity between IBD patients in terms of various biochemical features of disease activity and nutritional status. A total of 48 participants were enrolled, including 36 patients with IBD and 12 controls. We identified four main meaningful clusters of parameters. The first cluster included all SCFAs with strong mutual correlations. The second cluster contained red blood cell parameters and albumin levels. The third cluster included proinflammatory parameters such as tumor necrosis factor-α, CRP, platelets, and phosphoric, succinic, and lactic acids. The final cluster revealed an association between zonulin and interleukins IL-10, IL-17, and IL-22. Moreover, we observed an inverse correlation between IL-6 and body mass index. Our findings suggest a link between nutritional status, diet, and inflammatory parameters in patients with IBD, which contribute to a better adjustment of the nutritional treatment.

Association between fecal levels of Short-Chain Fatty Acids and serum Pro- and Anti-Inflammatory Cytokines in patients with Inflammatory Bowel Disease.

INTRODUCTION: Inflammatory bowel disease (IBD) represents a group of chronic inflammatory disorders characterized by dysbiosis and altered short-chain fatty acid (SCFA) level. The association between individual SCFA levels and cytokine levels is unknown. OBJECTIVES: We aimed to determine the fecal SCFA levels in patients with IBD in relation to disease severity and the serum levels of pro- and anti-inflammatory cytokines. PATIENTS AND METHODS: The study included 61 patients with IBD (inactive, 22; active, 39) and 16 controls. Fecal levels of organic acids (acetic, lactic, propionic, butyric, isovaleric, isobutyric, and valeric), serum levels of tumor necrosis factor-alpha (TNF-α), interleukin-10 (IL-10), IL-17, and IL-22, complete blood count and C-reactive protein (CRP) were measured. RESULTS: Patients with active IBD had reduced butyric, acetic, valeric, and isovaleric acid levels and elevated lactic acid levels in stool. Hemoglobin levels were positively correlated with the levels of acetic and butyric acids (R = 0.266 and R = 0.346, respectively; P <0.05). In addition, CRP levels were inversely correlated with butyric acid levels (R = -0.573; P <0.05). Higher serum TNF-α levels were observed in patients with active IBD compared with controls (6.64 pg/ml vs 2.05 pg/ml, P <0.05). No relationship was noted between the SCFA profile and cytokine levels. CONCLUSIONS: The study showed that determination of SCFA levels can be used to evaluate the activity of IBD. The relationship between individual SCFA and cytokine levels seems to be complex and requires further studies.

Determination of Essential Minerals and Trace Elements in Edible Sprouts from Different Botanical Families-Application of Chemometric Analysis.

BACKGROUND: elemental deficiency may result in the malfunctioning of human organisms. Sprouts, with their attractive looks and well-established popularity, may be considered as alternative sources of elements in the diet. Moreover, the uptake of micro- and macronutrients from sprouts is better when compared to other vegetable sources. The aim of the study was to determine and compare the level of the selected essential minerals and trace elements in 25 sprouts from different botanical families, to preselect the richest species of high importance for human diets. METHODS: the Cu, Zn, Mn, Fe, Mg, Ca determinations were performed using atomic absorption spectrometry with flame atomization and iodine by the colorimetric method. RESULTS: beetroot sprouts had the highest levels of Zn, Fe, and Mg, while onion sprouts were the richest in Mn and Ca, among all of the tested sprouts. Sprouts of the Brassicaceae family were generally richer in Ca, Mg, and Zn than sprouts from the Fabaceae family. Results allow preselection of the most perspective sprouts as possible dietary sources of essential minerals and trace elements. For rucola, leeks, onions, and beetroot sprouts, the data on minerals and trace element compositions were performed for the first time.

Association between Fecal Short-Chain Fatty Acid Levels, Diet, and Body Mass Index in Patients with Inflammatory Bowel Disease.

Disturbances in the production of bacterial metabolites in the intestine have been reported in diseases associated with dysbiosis, such as inflammatory bowel diseases (IBDs) that include two conditions: Crohn disease (CD) and ulcerative colitis (UC). Short-chain fatty acids (SCFAs) are the main dietary-fiber-derived bacterial metabolites associated with the course of intestinal inflammation. In this study, we assessed the relationship between body mass index (BMI), the type of diet used, and changes in fecal SCFA levels in patients with IBD. We performed nutritional assessments using a nutritional questionnaire and determined fecal SCFA levels in 43 patients with UC, 18 patients with CD, and 16 controls. Our results revealed that subjects with a BMI > 24.99 kg/m2 had higher levels of isobutyric acid, whereas those with a BMI < 18.5 kg/m2 had lower level of butyric, isovaleric, and propionic acids. Furthermore, we observed higher levels of valeric acid in controls than in IBD patients. We did not reveal a relationship between a specific SCFA and the type of diet, but eating habits appear to be related to the observed changes in the SCFA profile depending on BMI. In conclusion, we demonstrated that BMI is associated with SCFA levels in patients with IBD.

Fecal Levels of Lactic, Succinic and Short-Chain Fatty Acids in Patients with Ulcerative Colitis and Crohn Disease: A Pilot Study.

Intestinal dysbiosis plays a crucial role in the development of inflammatory bowel disease (IBD), including Crohn's disease (CD) and ulcerative colitis (UC). The importance of bacterial metabolites, such as short-chain fatty acids (SCFAs), succinic and lactic acids, as well as environmental factors that affect their production in the course of IBD, remains unclear. The aim of this study was to evaluate a profile of organic acids in the stool of CD and UC patients with different disease activity. We assessed a correlation between used medications, patient's diet, and SCFA levels. A total of 35 adult patients were included in the study. We did not observe significant differences in the levels of organic acids between the CD and UC groups, irrespective of disease activity, and a control group. However, propionic acid levels were higher in IBD patients who received trimebutine vs. those who did not (p = 0.031). Higher isobutyric acid levels were observed in patients treated with biological drugs compared with those without such treatment (p = 0.014). No significant correlations were found between organic acid levels and the frequency of dietary fiber consumption. Our results reveal a new link between medication use and organic acid levels in patients with IBD.

Phosphorus Solubilization by Bacillus Species.

Microbial solubilization applies the natural ability of a microorganism to liberate phosphorus from unavailable structures. The main mechanism recognized to be responsible for the solubilization of phosphorus is the production of different types of organic acids. Three kinds of Bacillus species and three types of raw materials (poultry bones, fish bones, and ash) were tested for solubilization. The following parameters were compared for all discussed cases: pH, specific growth rate, solubilization factor, released phosphorus concentration, and total and individual concentration of organic acids. Utilization of ash brought about the highest specific and maximum specific growth rates. A decrease in pH was observed in most of the discussed cases with the exception of fish bones. At the same time, fish bones had the highest concentration of released P2O5 and the highest total concentration of produced organic acids (gluconic, lactic, acetic, succinic, and propionic) in all discussed cases. The tested Bacillus species produced the mentioned acids with the exception of B. megaterium, where propionic acid was not present. The lactic and acetic acids were those produced in the highest amount. The kind of raw materials and type of Bacillus species used in solubilization had a strong influence on the kind of organic acids that were detected in the broth culture and its total concentration, which had a direct influence on the amount of released phosphorus. The combination of Bacillus megaterium with the fish bones at 5 g/L is proposed as the pair that gives the highest concentration of released phosphorus (483 ± 5 mg/L).

Link between Food Energy Density and Body Weight Changes in Obese Adults.

Regulating the energy density of food could be used as a novel approach for successful body weight reduction in clinical practice. The aim of this study was to conduct a systemic review of the literature on the relationship between food energy density and body weight changes in obese adults to obtain solid evidence supporting this approach. The search process was based on the selection of publications in the English language listed in public databases. A meta-analysis was performed to combine individual study results. Thirteen experimental and observational studies were identified and included in the final analysis. The analyzed populations consist of 3628 individuals aged 18 to 66 years. The studies varied greatly in terms of study populations, study design and applied dietary approaches. The meta-analysis revealed a significant association between low energy density foods and body weight reduction, i.e., -0.53 kg when low energy density foods were eaten (95% CI: -0.88, -0.19). In conclusions, this study adds evidence which supports the energy density of food as a simple but effective measure to manage weight in the obese with the aim of weight reduction.

Procedure optimization for extracting short-chain fatty acids from human faeces.

Short-chain fatty acids play an important role in the physiology and metabolism of the colon. Disturbed balance of such compounds in human gut can significantly contribute to etiological factors of various gastrointestinal disorders and it may also increase the risk of developing cancer or cardiovascular diseases. The aim of the study was to select the optimal parameters for acetic, propionic and butyric acids extraction from stool samples. The experimental conditions were optimized with respect to the solvent sample shacking time, sample ultrasounds (Ultrasound-Assisted Extraction, UAE) exposure time and the number of extractions from the particulate stool samples. The screening of experimental parameters was conducted with fractional factorial design of experiments, namely 3(3-1). The optimal conditions for UAE were found, namely ultrasound digestion time of 40 min (at 35°C), shaking time of 4 min, and the three subsequent extractions.

Cornelian cherry (cornus MAS L.) juice as a potential source of biological compounds.

Cornelian cherry (Cornus mas L.), apple (Malus domestica Borkh. cultivars Jonagold and Red Boskoop), pear (Pyrus communis cv. Conference), and plum (Prunus domestica L. cv. Common Plum) juices were assayed for their quality, organic acids, polyphenol content, and antioxidant activity (determined as a ferric-reducing antioxidant power [FRAP]). Cornelian cherry juice revealed the highest titratable acidity-2.58 g malic acid/100 g. The highest polyphenol content was also noted for Cornelian cherry-45.6 mg gallic acid (GA)/g. The medium level of polyphenols was proved for Jonagold apple (22.8 mg GA1/g). The lowest level was recorded for Common Plum (9.60 mg GA/g), followed by Conference pear and Red Boskoop apple (12.3 and 21.3 mg GA/g, respectively). The FRAP values were 1.97 mmol Fe/L for Common Plum juice, 2.37 mmol Fe/L for Conference pear juice, and 3.92 mmol Fe/L for both Red Boskoop and Jonagold apple juices. However, the obtained data indicated outstanding antioxidant properties for Cornelian cherry juice. In this case, FRAP reached 23.5 mmol Fe/L. The calculated correlation coefficient FRAP versus polyphenols indicates that the antioxidant status for Cornelian cherry is not only correlated with polyphenols but also with other biological compounds. The obtained results indicated that Cornelian cherry is a valuable source of substances with a high antioxidant activity.