Alanine, a potential amino acid biomarker of pediatric sepsis: a pilot study in PICU.

Sepsis is characterized by a metabolic disorder of amino acid occurs in the early stage; however, the profile of serum amino acids and their alterations associated with the onset of sepsis remain unclear. Thus, our objective is to identify the specific kinds of amino acids as diagnostic biomarkers in pediatric patients with sepsis. Serum samples were collected from patients with sepsis admitted to the pediatric intensive care unit (PICU) between January 2019 and December 2019 on the 1st, 3rd and 7th day following admission. Demographic and laboratory variables were also retrieved from the medical records specified times. Serum amino acid concentrations were detected by UPLC-MS/MS system. PLS-DA (VIP > 1.0) and Kruskal-Wallis test (p < 0.05) were employed to identify potential biomarkers. Spearman's rank correlation analysis was conducted to find the potential association between amino acid levels and clinical features. The diagnostic utility for pediatric sepsis was assessed using receiver operating characteristic (ROC) curve analysis. Most of amino acid contents in serum were significantly decreased in patients with sepsis, but approached normal levels by the seventh day post-diagnosis. Threonine (THR), lysine (LYS), valine (VAL) and alanine (ALA) emerged as potential biomarkers related for sepsis occurrence, though they were not associated with PELOD/PELOD-2 scores. Moreover, alterations in serum THR, LYS and ALA were linked to complications of brain injury, and serum ALA levels were also related to sepsis-associated acute kidney injury. Further analysis revealed that ALA was significantly correlated with the Glasgow score, serum lactate and glucose levels, C-reactive protein (CRP), and other indicators for liver or kidney dysfunction. Notably, the area under the ROC curve (AUC) for ALA in distinguishing sepsis from healthy controls was 0.977 (95% CI: 0.925-1.000). The serum amino acid profile of children with sepsis is significantly altered compared to that of healthy controls. Notably, ALA shows promise as a potential biomarker for the early diagnosis in septic children.

Acute effects of oral mesna administration on the full amino acid profile and 3-methylhistidine: secondary results from the CYLOB dose-finding study.

Plasma total cysteine (tCys) is strongly associated with fat mass in humans. Mesna lowers plasma tCys in a dose-dependent manner, but it is not known whether it interferes with metabolism of other amino acids or protein. In this Phase-1 study, we show that a single dose of mesna administered at 400, 800, 1200 or 1600 mg to 6-7 individuals per dose only slightly affects amino acid profiles, with increases in plasma valine across dose levels. There were no effects of mesna on 3-methylhistidine, a marker of protein breakdown.

Effect of source and frequency of rumen-protected protein supplementation on mammary gland amino acid metabolism and nitrogen balance of dairy cattle.

The AA profile of MP affects mammary gland metabolism and milk N efficiency of dairy cattle. Further, the frequency of dietary protein supplementation may influence N partitioning leading to reduced N excretion. This study investigated the effect of source and frequency of rumen-protected (RP) protein supplementation on apparent total-tract digestibility, milk production, mammary gland AA metabolism, and N balance of dairy cattle. Twenty-eight Holstein-Friesian cows (2.3 ± 0.9 lactations; 93 ± 27 DIM; mean ± SD) were used in a randomized complete block design and fed a basal TMR consisting of 41% corn silage, 32% grass silage, and 27% concentrate (DM basis) and formulated to meet 100% and 95% of net energy and MP requirements, respectively. Cows were adapted to the basal TMR in a freestall barn for 7 d, moved to individual tiestalls for 13 d of adaptation to dietary treatments, and then moved into climate respiration chambers for a 4-d measurement period. Treatments consisted of the basal TMR (CON; 159 g CP/kg DM) or the basal TMR including 1 of 3 iso-MP supplements: (1) 315-g mixture of RP soybean meal and RP rapeseed meal fed daily (ST-RPSR), (2) 384-g mixture of RP His, RP Lys, and RP Met fed daily (ST-RPAA), and (3) 768-g mixture of RP His, RP Lys, and RP Met fed every other day (OS-RPAA). The basal TMR with the addition of treatment supplements was designed to deliver 100% of required MP over a 48-h period. The mixture of His, Lys, and Met was formulated to deliver digestible AA in amounts relative to their concentration in casein. Compared with ST-RPSR, ST-RPAA increased milk protein and fat concentration, increased the arterial concentration of total His, Lys, and Met (HLM), decreased mammary clearance of HLM, and increased clearance of Phe, Leu, and Tyr (tendency for Leu and Tyr). Rumen-protected protein source did not affect N balance, but the marginal use efficiency (efficiency of transfer of RP protein supplement into milk protein) of ST-RPAA (67%) was higher than that of ST-RPSR (17%). Milk protein concentration decreased with OS-RPAA compared with ST-RPAA. Arterial concentration of HLM increased on the nonsupplemented day compared with the supplemented day with OS-RPAA, and there was no difference in arterial HLM concentration across days with ST-RPAA. Mammary uptake of HLM tended to increase on the nonsupplemented day compared with the supplemented day with OS-RPAA. Supplementation frequency of RP AA did not affect N balance or overall milk N efficiency, but the marginal use efficiency of OS-RPAA (49%) was lower compared with ST-RPAA. Overall, mammary glands responded to an increased supply of His, Lys, and Met by reducing efflux of other EAA when RP His, RP Lys, and RP Met were supplemented compared with RP plant proteins. Mammary glands increased sequestration of EAA (primarily HLM) on the nonsupplemented day with OS-RPAA, but supplementing RP AA according to a 24-h oscillating pattern did not increase N efficiency over static supplementation.

Quality Assessment of Reconstructed Cow, Camel and Mare Milk Powders by Near-Infrared Spectroscopy and Chemometrics.

Milk powders are becoming a major attraction for many industrial applications due to their nutritional and functional properties. Different types of powdered milk, each with their own distinct chemical compositions, can have different functionalities. Consequently, the development of rapid monitoring methods is becoming an urgent task to explore and expand their applicability. Lately, there is growing emphasis on the potential of near-infrared spectroscopy (NIRS) as a rapid technique for the quality assessment of dairy products. In the present work, we explored the potential of NIRS coupled with chemometrics for the prediction of the main functional and chemical properties of three types of milk powders, as well as their important processing parameters. Mare, camel and cow milk powders were prepared at different concentrations (5%, 10% and 12%) and temperatures (25 °C, 40 °C and 65 °C), and then their main physicochemical attributes and NIRS spectra were analyzed. Overall, high accuracy in both recognition and prediction based on type, concentration and temperature was achieved by NIRS-based models, and the quantification of quality attributes (pH, viscosity, dry matter content, fat content, conductivity and individual amino acid content) also resulted in high accuracy in the models. R2CV and R2pr values ranging from 0.8 to 0.99 and 0.7 to 0.98, respectively, were obtained by using PLSR models. However, SVR models achieved higher R2CV and R2pr values, ranging from 0.91 to 0.99 and 0.80 to 0.99, respectively.

Nutritional and physicochemical characteristics of Asiatic hard clam powder prepared by different cook-drying processes: a comparative study.

BACKGROUND: Asiatic hard clam (Meretrix meretrix) is an underutilized bivalve resource. This study discusses dried clam powders prepared from this resource to enhance its utilization and improve nutritional security in protein-deficient populations. Dried clam powder was prepared from Asiatic hard clam and the effects of different pre-cooking methods (boil-dried clam powder, BDCP; steam-dried clam powder, SDCP; and microwave-dried clam powder, MDCP) on nutritional (proximate composition, amino acid profiling, mineral profiling, fatty acid profiling) and physicochemical qualities were investigated. RESULTS: Different pre-cooking methods significantly influenced the characteristics of the clam powder. The MDCP sample showed the highest concentration of amino acids, polyunsaturated fatty acids, Na, K, Ca and Mg content compared to BDCP and SDCP. The boiling process led to a loss of nutritional quality in terms of amino acids and macrominerals. The MDCP displayed the highest solubility in water (30.10%) but its oil and water absorption characteristics were the lowest among all the samples. Boil-cooked clam powder displayed the highest oil binding (2.03 mL g-1 protein) capacity. Boiling and steaming processes resulted in malondialdehyde generation compared to microwaving. Different pre-cooking processes did not influence the colour attributes significantly, but the control sample prepared without pre-cooking (CCP) had a significantly lower L* value (32.34), resulting in a darker product. In vitro digestibility of the clam powder varied in the order MDCP > SDCP > BDCP > CCP. CONCLUSION: The study demonstrated that nutritionally rich protein powder can be prepared from Asiatic hard clam. Based on the results, microwave pre-cooking is considered the best pre-cooking method to preserve the nutritional qualities of clam powder. © 2024 Society of Chemical Industry.

A specific plasma amino acid profile in the Insulin2Q104del Kuma mice at the diabetic state and reversal from hyperglycemia.

The metabolites in the plasma serve as potential biomarkers of disease. We previously established an early-onset diabetes mouse model, Ins2+/Q104del Kuma mice, under a severe immune-deficient (Rag-2/Jak3 double-deficient in BALB/c) background. Here, we revealed the differences in plasma amino acid profiles between Kuma and the wild-type mice. We observed an early reduction in glucogenic and ketogenic amino acids, a late increase in branched-chain amino acids (BCAAs) and succinyl CoA-related amino acids, and a trend of increasing ketogenic amino acids in Kuma mice than in the wild-type mice. Kuma mice exhibited hyperglucagonemia at high blood glucose, leading to perturbations in plasma amino acid profiles. The reversal of blood glucose by islet transplantation normalized the increases of the BCAAs and several aspects of the altered metabolic profiles in Kuma mice. Our results indicate that the Kuma mice are a unique animal model to study the link between plasma amino acid profile and the progression of diabetes for monitoring the therapeutic effects.

Common alterations in plasma free amino acid profiles and gut microbiota-derived tryptophan metabolites of five types of cancer patients.

Amino acids not only play a vital role in the synthesis of biological molecules such as proteins in cancer malignant cells, they are also essential metabolites for immune cell activation and antitumor effects in the tumor microenvironment. The abnormal changes in amino acid metabolism are closely related to the occurrence and development of tumors and immunity. Intestinal microorganisms play an essential role in amino acid metabolism, and tryptophan and its intestinal microbial metabolites are typical representatives. However, it is known that the cyclic amino acid profile is affected by specific cancer types, so relevant studies mainly focus on one type of cancer and rarely study different cancer forms at the same time. The objective of this study was to examine the PFAA profile of five cancer patients and the characteristics of tryptophan intestinal microbial metabolites to determine whether there are general amino acid changes across tumors. Plasma samples were collected from esophageal (n = 53), lung (n = 73), colorectal (n = 94), gastric (n = 55), breast cancer (n = 25), and healthy control (HC) (n = 139) subjects. PFAA profile and tryptophan metabolites were measured, and their perioperative changes were examined using high-performance liquid chromatography. Univariate analysis revealed significant differences between cancer patients and HC. Furthermore, multivariate analysis discriminated cancer patients from HC. Regression diagnosis models were established for each cancer group using differential amino acids from univariate analysis. Receiver-operating characteristic analysis was applied to evaluate these diagnosis models. Finally, GABA, arginine, tryptophan, taurine, glutamic acid, and melatonin showed common alterations across all types of cancer patients. Metabolic pathway analysis shows that the most significant enrichment pathways were tryptophan, arginine, and proline metabolism. This study provides evidence that common alterations of the metabolites mentioned above suggest their role in the pathogenesis of each cancer patient. It was suggested that multivariate models based on PFAA profiles and tryptophan metabolites might be applicable in the screening of cancer patients.

Plasma free amino acid profiles are associated with serum high molecular weight adiponectin levels in Japanese medical check-up population without type 2 diabetes mellitus.

To prevent the progression of type 2 diabetes mellitus (T2DM), early detection and intervention are important. Several studies have already shown that the serum adiponectin level could be useful for evaluating the future risk of T2DM. Recently, plasma free amino acid (PFAA) concentrations have also emerged as potential biomarkers that predict the future onset of T2DM. In this study, we aimed to further characterise PFAA profiles by elucidating the association with the serum high molecular weight (HMW) adiponectin level in this cross-sectional study. A total of 1000 Japanese subjects who underwent medical check-ups were enrolled, and their plasma concentrations of 21 amino acids and clinical parameters were measured. The subjects without T2DM were divided into quartiles (Q1-4) by serum HMW adiponectin level, and the association with between PFAA concentrations was analysed. Concentrations of glutamate, alanine, proline, tyrosine, histidine, methionine, lysine, branched-chain amino acids (BCAAs) and tryptophan varied significantly according to the adiponectin quartile. Furthermore, serum adiponectin levels showed significant inverse correlations with these amino acids. The change in the PFAA profile in the group with the lowest adiponectin concentrations (Q1) was similar to that of T2DM patients. Although both adiponectin levels and PFAA concentrations are known to be altered by the accumulation of visceral fat and insulin resistance, the levels of glutamate, BCAA, lysine and tryptophan remain significantly associated with adiponectin level after adjustment for age, body mass index and homeostasis model assessment of insulin resistance, showing the direct association between PFAA concentrations and the serum HMW adiponectin level. Registration number: University Hospital Medical Information Network Clinical Trials Registry (UMIN-CTR) UMIN000029920, registered on Nov 13th 2017 (prospectively registered).

The nutritional composition and cell size of microbial biomass for food applications are defined by the growth conditions.

BACKGROUND: It is increasingly recognized that conventional food production systems are not able to meet the globally increasing protein needs, resulting in overexploitation and depletion of resources, and environmental degradation. In this context, microbial biomass has emerged as a promising sustainable protein alternative. Nevertheless, often no consideration is given on the fact that the cultivation conditions affect the composition of microbial cells, and hence their quality and nutritional value. Apart from the properties and nutritional quality of the produced microbial food (ingredient), this can also impact its sustainability. To qualitatively assess these aspects, here, we investigated the link between substrate availability, growth rate, cell composition and size of Cupriavidus necator and Komagataella phaffii. RESULTS: Biomass with decreased nucleic acid and increased protein content was produced at low growth rates. Conversely, high rates resulted in larger cells, which could enable more efficient biomass harvesting. The proteome allocation varied across the different growth rates, with more ribosomal proteins at higher rates, which could potentially affect the techno-functional properties of the biomass. Considering the distinct amino acid profiles established for the different cellular components, variations in their abundance impacts the product quality leading to higher cysteine and phenylalanine content at low growth rates. Therefore, we hint that costly external amino acid supplementations that are often required to meet the nutritional needs could be avoided by carefully applying conditions that enable targeted growth rates. CONCLUSION: In summary, we demonstrate tradeoffs between nutritional quality and production rate, and we discuss the microbial biomass properties that vary according to the growth conditions.

Optimization of Heterotrophic Culture Conditions for the Microalgae Euglena gracilis to Produce Proteins.

Euglena gracilis is one of the few permitted edible microalgae. Considering consumer acceptance, E. gracilis grown heterotrophically with yellow appearances have wider food industrial applications such as producing meat analogs than green cells. However, there is much room to improve the protein content of heterotrophic culture cells. In this study, the effects of nitrogen sources, temperature, initial pH, and C/N ratios on the protein production of E. gracilis were evaluated under heterotrophic cultivation. These results indicated that ammonium sulfate was the optimal nitrogen source for protein production. The protein content of E. gracilis cultured by ammonium sulfate increased by 113% and 44.7% compared with that cultured by yeast extract and monosodium glutamate, respectively. The manipulation of the low C/N ratio further improved E. gracilis protein content to 66.10% (w/w), which was 1.6-fold of that in the C/N = 25 group. Additionally, amino acid analysis revealed that the nitrogen-to-protein conversion factor (NTP) could be affected by nitrogen sources. A superior essential amino acid index (EAAI) of 1.62 and a balanced amino acid profile further confirmed the high nutritional value of E. gracilis protein fed by ammonium sulfate. This study highlighted the vast potency of heterotrophic cultured E. gracilis as an alternative dietary protein source.

Amino Acid Profiling of Follicular Fluid in Assisted Reproduction Reveals Important Roles of Several Amino Acids in Patients with Insulin Resistance.

The global prevalence of insulin resistance (IR) is increasing continuously, influencing metabolic parameters and fertility. The metabolic changes due to IR can alter the molecular composition of plasma and other body fluids. Follicular fluid (FF) is derived mainly from plasma, and it is a critical microenvironment for the developing oocytes. It contains various metabolites and amino acids, and the quality of the oocytes is linked at least partially to amino acid metabolism. Our goal was to quantitatively determine the amino acid (AA) profile of FF in IVF patients and to compare IR and non-insulin resistance (NIR) groups to investigate the AA changes in their FF. Using UHPLC-based methods, we quantified the main 20 amino acids from human FF samples in the IR and NIR groups. Several amino acids (aspartate, glycine, glutamate, and cysteine) differed significantly (p < 0.05 or less) between the two groups. The most significant alterations between the IR and NIR groups were related to the glutathione metabolic pathway involving glycine, serine, and threonine. Since insulin resistance alters the amino acid composition of the FF, the oocytes may undergo metabolism-induced changes resulting in poor oocyte quality and less fertility in the insulin resistance groups.

Solid-state fermentation improves the quality of chrysanthemum waste as an alternative feed ingredient.

Chrysanthemum waste (CW) is an agricultural and industrial by-product produced during chrysanthemum harvesting, drying, preservation, and deep processing. Although it is nutritious, most CW is discarded, wasting resources and contributing to serious environmental problems. This work explored a solid-state fermentation (SSF) strategy to improve CW quality for use as an alternative feed ingredient. Orthogonal experiment showed that the optimal conditions for fermented chrysanthemum waste (FCW) were: CW to cornmeal mass ratio of 9:1, Pediococcus cellaris + Candida tropicalis + Bacillus amyloliquefaciens proportions of 2:2:1, inoculation amount of 6%, and fermentation time of 10 d. Compared with the control group, FCW significantly increased the contents of crude protein, ether extract, crude fiber, acid detergent fiber, neutral detergent fiber, ash, calcium, phosphorus, and total flavonoids (p < 0.01), and significantly decreased pH and saponin content (p < 0.01). SSF improved the free and hydrolyzed amino acid profiles of FCW, increased the content of flavor amino acids, and improved the amino acid composition of FCW protein. Overall, SSF improved CW nutritional quality. FCW shows potential use as a feed ingredient, and SSF helps reduce the waste of chrysanthemum processing.

Antioxidant potential and amino acid profile of ultrafiltration derived peptide fractions of spent hen meat protein hydrolysate.

Spent hen meat is considered as a category of waste generated by the poultry sector which can lead to serious environmental concerns if not disposed and utilized properly. In this work, spent hen meat was hydrolysed by 2% Flavourzyme (6.5 pH, 55 °C) followed by ultrafiltration to produce three peptide fractions with molecular weights > 10 kDa, 5-10 kDa and < 5 kDa. These fractions were evaluated for antioxidant potential, SDS PAGE and amino acid profile. The SDS PAGE profile demonstrated bands in the low molecular weight (< 10 kDa) region. Peptide fractions of < 5 kDa exhibited highest antioxidant activity and, essential as well as hydrophobic amino acid composition than whole hydrolysate and other peptide fractions. Incorporation of the identified hydrolysate fraction in food could improve its shelf stability while serving as a preventive component against human degenerative diseases.

Comparison of Physicochemical and Structural Properties of Acid-Soluble and Pepsin-Soluble Collagens from Blacktip Reef Shark Skin.

Fish collagen has been widely used in tissue engineering (TE) applications as an implant, which is generally transplanted into target tissue with stem cells for better regeneration ability. In this case, the success rate of this research depends on the fundamental components of fish collagen such as amino acid composition, structural and rheological properties. Therefore, researchers have been trying to find an innovative raw material from marine origins for tissue engineering applications. Based on this concept, collagens such as acid-soluble (ASC) and pepsin-soluble (PSC) were extracted from a new type of cartilaginous fish, the blacktip reef shark, for the first time, and were further investigated for physicochemical, protein pattern, microstructural and peptide mapping. The study results confirmed that the extracted collagens resemble the protein pattern of type-I collagen comprising the α1, α2, ß and γ chains. The hydrophobic amino acids were dominant in both collagens with glycine and hydroxyproline as major amino acids. From the FTIR spectra, α helix (27.72 and 26.32%), ß-sheet (22.24 and 23.35%), ß-turn (21.34 and 22.08%), triple helix (14.11 and 14.13%) and random coil (14.59 and 14.12%) structures of ASC and PSC were confirmed, respectively. Collagens retained their triple helical and secondary structure well. Both collagens had maximum solubility at 3% NaCl and pH 4, and had absorbance maxima at 234 nm, respectively. The peptide mapping was almost similar for ASC and PSC at pH 2, generating peptides ranging from 15 to 200 kDa, with 23 kDa as a major peptide fragment. The microstructural analysis confirmed the homogenous fibrillar nature of collagens with more interconnected networks. Overall, the preset study concluded that collagen can be extracted more efficiently without disturbing the secondary structure by pepsin treatment. Therefore, the blacktip reef shark skin could serve as a potential source for collagen extraction for the pharmaceutical and biomedical applications.

Plasma essential amino acid concentration and profile are associated with performance of lactating dairy cows as revealed through meta-analysis and hierarchical clustering.

Our aim was to explore whether changes in plasma essential AA (EAA) concentration ([EAA]p) or profile (defined here as the molar proportion of individual [EAA]p relative to the total [EAA]p) may serve as an indicator of the EAA status of a cow. We undertook a meta-analysis with the objectives to determine if different plasma EAA profiles exist among cows and to explore the association of [EAA]p or the profile of EAA with lactating cow performance and measures of N utilization. We hypothesized the existence of differences in [EAA]p and different plasma EAA profile for cows with greater milk output, feed efficiency, and greater N use efficiency (NUE; milk true protein-N:N intake) compared with cows with lower milk output, feed efficiency, and lower NUE. The data set included 22 feeding trials and 96 dietary treatments. First, a mixed-effect model analysis was used to predict [EAA]p in response to the categorical fixed effect of EAA, continuous fixed effect of National Research Council model-predicted metabolizable protein (MP) supply, continuous fixed effect of body weight, the fixed effect of EAA and MP supply interaction, the fixed effect of EAA and body weight interaction, and the random effect of study. Then, residuals of the model were standardized based on Z-score and clustered using the hierarchical method (Euclidean distance and Ward's minimum variance method) resulting in 2 clusters. Finally, a fixed-effect model was used to evaluate the significance with which clusters were associated with [EAA]p, cow performance, feed efficiency, and NUE. The total concentration of [EAA]p was lower (784 vs. 983 µM) and the concentration of each EAA was on average 22 µM lower for cows in cluster 1 compared with cluster 2 with the smallest and greatest difference found for Met (4 µM) and Val (59 µM), respectively. The percentage difference in [EAA]p was the smallest for Thr (-5.3%) and the greatest for Leu (-37.1%). There was no difference between clusters for Arg, His, and Met molar proportions; however, cows in cluster 1 had a lower molar proportion of Leu and a tendency for lower molar proportion of Val compared with cows in cluster 2. Additionally, cows in cluster 1 had greater molar proportions of Ile, Lys, and Thr and a tendency for greater molar proportion of Phe compared with cows in cluster 2. The fixed-effect model analysis indicated that cows in cluster 1 had higher milk energy output (+3.2 Mcal/d), true protein yield (+87 g/d) and fat yield (+236 g/d), feed efficiency (milk Mcal:dry matter intake; +8% unit), and a tendency for greater MP efficiency (Milk true protein/MP supply; +2.3% unit) than cows in cluster 2. These results suggested greater use of EAA by the mammary gland (as reflected by greater milk protein synthesis) and lower hepatic catabolism of AA (as reflected by a tendency to greater MP efficiency) in cows of cluster 1 compared with cluster 2. Our findings should be evaluated further, including whether the relative molar proportions of plasma EAA might serve as a holistic indicator of the EAA status of cows as related to their productivity, feed efficiency and N utilization.

Lactational performance of dairy cows in response to supplementing N-acetyl-l-methionine as source of rumen-protected methionine.

The objective of this experiment was to evaluate the effects of supplementing a rumen-protected source of Met, N-acetyl-l-methionine (NALM), on lactational performance and nitrogen metabolism in early- to mid-lactation dairy cows. Sixty multiparous Holstein dairy cows in early lactation (27 ± 4.3 d in milk, SD) were assigned to 4 treatments in a randomized complete block design. Cows were blocked by actual milk yield. Treatments were as follows: (1) no NALM (control); (2) 15 g/d of NALM (NALM15); (3) 30 g/d of NALM (NALM30); and (4) 45 g/d of NALM (NALM45). Diets were formulated using a Cornell Net Carbohydrate and Protein System (CNCPS) v.6.5 model software to meet or exceed nutritional requirements of lactating dairy cows producing 42 kg/d of milk and to undersupply metabolizable Met (control) or supply incremental amounts of NALM. The digestible Met (dMet) supply for control, NALM15, NALM30, and NALM45 were 54.7, 59.8, 64.7, and 72.2 g/d, respectively. The supply of dMet was 88, 94, 104, and 115% of dMet requirement for control, NALM15, NALM30, and NALM45, respectively. Milk yield data were collected, dry matter intake (DMI) was measured daily, and milk samples were collected twice per week for 22 wk. Blood, ruminal fluid, urine, and fecal samples were collected during the covariate period and during wk 4, 8, and 16. Data were analyzed using the GLIMMIX procedure of SAS (SAS Institute) using covariates in the model for all variables except body weight. Linear, quadratic, and cubic contrasts were also tested. Treatments did not affect DMI, milk yield, and milk component concentration and yield; however, feed efficiency expressed as milk yield per DMI and 3.5% fat-corrected milk per DMI were quadratically affected, with greater response observed for NALM15 and NALM30 compared with control. Acetate proportion linearly increased, whereas propionate proportion linearly decreased with NALM supplementation. Blood urea nitrogen linearly decreased with NALM supplementation. Total plasma essential AA concentrations were quadratically affected, as greater values were observed for control and NALM45 than other treatments. Plasma Met concentration was quadratically affected as lower levels were observed with NALM15, whereas Met concentrations increased with NALM45 compared with control. Nitrogen utilization efficiency and apparent total-tract nutrient digestibility were not affected by treatment. Supplementation of NALM at 15 or 30 g/head per day resulted in the greatest improvements in feed efficiency without affecting N metabolism of early- to mid-lactation dairy cows.

Chemical Composition, Nutritional Value, and Acceptance of Nut Bars with the Addition of Edible Insect Powder.

Six types of nut-based bars with the addition of edible insect flour were obtained. Flours made from three different insects (Tenebrio molitor L., Acheta domesticus L., Alphitobius diaperinus P.) were used at two different additive levels (15% and 30%) in relation to the weight of the nuts. The addition of insect flour significantly increased protein content and the insoluble fraction of dietary fiber. The largest amount of these compounds was found in bars with 30% cricket flour, 15.51 g/100 g and 6.04 g/100 g, respectively, in comparison to standard bars, 10.78 g/100 g and 3.14 g/100 g, respectively. The greatest consumer acceptance was found in relation to bars with buffalo worm flour. The overall acceptance of these bars was 6.26-6.28 points compared to 6.48 for standard bars. Bars and raw materials were characterized by the high biological value of the protein. Cis linoleic acid dominated among unsaturated fatty acids. The percentage of this compound was in the range of 69.56%, for bars with a 30% addition of buffalo worm flour, to 73.88%, for bars with 15% cricket flour. Instrumental analysis of taste and smell compounds showed the presence of compounds such as 3-methylbutanoic acid, hexanal, and 2,3-pentanedione.

Chemical Profiling and Nutritional Evaluation of Bee Pollen, Bee Bread, and Royal Jelly and Their Role in Functional Fermented Dairy Products.

Honeybee products, as multicomponent substances, have been a focus of great interest. The present work aimed to perform the nutritional and chemical profiling and biochemical characterization of bee pollen (BP), bee bread (BB), and royal jelly (RJ) and study their applications in the fortification of functional fermented dairy products. Their effects on starter cultures and the physicochemical and sensorial quality of products were monitored. A molecular networking analysis identified a total of 46 compounds in the three bee products that could be potential medicines, including flavonoids, fatty acids, and peptides. BB showed the highest protein and sugar contents (22.57 and 26.78 g/100 g), which cover 45.14 and 53.56% of their daily values (DVs), with considerable amounts of the essential amino acids threonine and lysine (59.50 and 42.03%). BP, BB, and RJ can be considered sources of iron, as 100 g can cover 141, 198.5, and 94.94% of DV%, respectively. BP was revealed to have the highest phenolic and flavonoid contents (105.68 and 43.91 µg/g) and showed a synergetic effect when mixed with RJ, resulting in increased antioxidant activity, while BB showed a synergetic effect when mixed with RJ in terms of both antioxidant and proteolytic powers (IC50 7.54, 11.55, 12.15, 12.50, and 12.65 cP compared to the control (10.55 cP)), reflecting their organoleptic properties and highlighting these health-oriented products as promising natural products for human health care.

Misdiagnosis and Mistreatment of a Rare Case of Intracranial Oncogenic Osteomalacia with an Altered Amino Acid Profile.

Background. Oncogenic osteomalacia (OO), also known as tumor-induced osteomalacia (TIO), is a rare paraneoplastic syndrome caused by mesechymal tumors secreting fibroblast growth factor 23 (FGF23). Common in middle age, these tumors are often disclosed by progressive generalized bone pain and muscle weakness, along with an altered biochemical profile. Despite its characteristic presentation, the disease is often underrecognized with delayed onset of surgical or pharmacological intervention that can have serious repercussions on the patients' health and quality of life. Case presentation. We describe the case of a 65-year-old Caucasian man presenting TIO with intracranial and spinal localizations and Fanconi-like aminoaciduria. The condition was misdiagnosed and mistreated for three years, leading to loss of self-sufficiency and depression. Following proper identification, the spinal mass was excised with complete remission of the functional symptoms. As it was not possible to remove the intracranial lesion, the patient was treated conservatively with calcitriol and phosphorous supplements that granted good metabolic control up to the time of a recent follow-up visit (at 5 years). Conclusions. The finding of an altered amino acid profile, not usually reported in these cases, should prompt clinicians to a wider usage of these molecules as suitable candidates for metabolic diseases. In addition to providing central information, they are easy to obtain and inexpensive to analyze. Such determination could help to speed up the diagnostic process, as a long-lasting history of misdiagnosis and mistreatments can lead primarily to clinical worsening, but also to the use of expensive, useless medications with side effects that contribute to poor patient health.

Effect of Moringa oleifera seed flour on the rheological, physico-sensory, protein digestibility and fatty acid profile of cookies.

In the present study, debittered Moringa Oleifera seed flour (DDMF) rich in protein, vitamins, minerals and balanced amino acid and fatty acid profile was used to develop functional cookies. DDMF was incorporated at 25, 50, 75 and 100% levels and studied their effect on flour rheological, physicochemical, micro-structural, sensory and nutritional properties of cookies. The results revealed that the addition of an increasing amount of DDMF from 0 to 100% increased water absorption (59.5-77%) by farinograph study; cookie dough hardness (89.2-284.7 N); decreased pasting temperature (60.2-30.1 °C) and peak viscosity (696-9 BU) by amylograph study. SEM studies of cookies indicated that, in control cookies, starch granules are completely gelatinized and enmeshed in the gluten protein matrix, whereas, in 50% DDMF incorporated cookies, partially gelatinized starch granules are seen embedded in a weak protein matrix. Sensory evaluation showed that incorporating DDMF, up to 50% of cookies had clean mouthfeel without any residue formation and were highly acceptable; however, beyond that limit, they became brittle. The addition of 50% DDMF increased cookies' in-vitro protein digestibility, mineral contents, and fatty acids content. Thus, the nutritional quality of cookies concerning quantity and quality of protein and fat could be enhanced by incorporating DDMF.