Hen Egg Research Articles

Genotype and dietary supplementation of ginger (Zingibar officinale) rhizome powder affect egg quality traits of layer chickens

This study evaluated the effects of ginger (Z. officinale) rhizome powder on egg qualities of aged commercial and native hens using 160 Shaver Brown (SB) and Nigerian heavy ecotype native hens (HEN). Experimental groups per genotype were control group or basal diet group, and ginger rhizome powder supplemented group (20 g/kg, ginger group). For each genotype, forty (40) hens were assigned to the control group and 120 hens to the ginger group. Each experimental group had four (4) replicates containing 10 hens each for the control groups, and 30 hens each for the ginger groups. The hens were housed individually in laying cages and fed the experimental diets at 125 g/bird/day. Water was given ad libitum and photoperiod was 12 h light and darkness. 80 eggs from SB and 40 eggs from HEN control groups, and 80 eggs/genotype from the ginger groups were used to evaluate egg qualities while 10 shell samples were used to evaluate shell mineral contents for each group. In addition, within genotypes, eggs belonging to the ginger groups were grouped into egg weight and egg shape index grades and assessed for egg qualities. SB eggs had significantly (p ≤ 0.05) higher weight (EW), length (EL), diameter (ED), surface area (ESA), volume (EV), and shape index (ESI); yolk, albumen, and shell weights (YW, AW and SW); albumen height, diameter, and ratio (AH, AD, and AR); shell volume (SV), compression fracture force (SCFF), and calcium content but lower egg specific gravity (ESG), yolk ratio (YR), yolk/albumen ratio (Y/A ratio), shell Zinc, and Phosphorus contents compared to HEN eggs. Ginger groups had higher EW, EL, ESA, EV, YW, YD, YR, AH, AI, Y/A ratio, and shell Zn but lower ESG, YH and YI compared to control groups. SB-ginger group had highest (p ≤ 0.000) AD while HEN-ginger group had highest (p ≤ 0.05) shell Zinc, and Phosphorus contents. EW grades differed significantly in EW, ESA, ESG, ESI and SCFF in the two genotypes while ESI grades differed significantly in ESI and Y/A ratio in SB but in ESI, YI and Haugh unit in HEN. In conclusion, genotype, diet, genotype x diet, EW and ESI grades influenced egg quality traits. These factors are important for egg quality evaluation and genetic improvement.

Development and Evaluation of Different Electrospun Cysteamine-Loaded Nanofibrous Webs: A Promising Option for Treating a Rare Lysosomal Storage Disorder.

Nanofibers can be utilized to overcome the challenges faced by conventional ophthalmic formulations. This study aimed to develop and characterize cysteamine (Cys)-loaded nanofiber-based ophthalmic inserts (OIs) as a potential candidate for the treatment of ophthalmic cystinosis using water-soluble polyvinyl alcohol (PVA)/poloxamer 407 (PO-407) and water-insoluble tetraethoxysilane (TEOS)/PVA nanofibers. Plain and Cys-loaded fibers in different proportions were prepared by the electrospinning method and studied for their morphological, physicochemical, release study, cytocompatibility effects, and stability study. The fiber formation was confirmed by scanning electron microscopy, while Fourier transform infrared spectra showed the most critical peaks for the Cys and the excipients. The release of the Cys was fast from the two polymeric matrices (≤20 min). The release from TEOS/PVA nanofibers is characterized by Case II transport (0.75 < β < 1), while the release from PVA/PO-407 nanofibers follows Fickian diffusion (β < 0.75). The cytocompatibility of compositions was confirmed by hen eggs tested on the chorioallantoic membrane (HET-CAM) of chick embryos. All formulations remained stable under stress conditions (40 ± 2 °C, 75 ± 5% relative humidity) regarding morphology and physicochemical characteristics. The developed nanofibrous mats could be an excellent alternative to available Cys drops, with better stability and convenience of self-administration as OIs.

Modification of the Protein Amino Acid Content in Hen Eggs as a Consequence of Different Concentrations of Lupine and Soy in Feed.

The effect of the diet modification (soybean and lupine addition) on the content of protein and amino acids (AA) in eggs was studied. Both the sampling day and the diet influenced the total protein content. In albumen, the lowest protein content (10.6%) was noted after administering a diet containing 25% lupine; in the same egg the yolk contained the most proteins (16.7%). In the content of nonessential AA (NAA) in egg yolks, differences were noted only for cysteine, with its the highest content in the yolks of the control group. The stable content of essential yolk amino acids (EAA) was observed only for isoleucine, leucine, tryptophan and phenylalanine. The highest contents of EAA and NAA were recorded in the yolks of the control group (~47 and ~53 g/100 g of protein, respectively) and in the group with 25% additions of lupine (~42 and ~51 g/100 g of protein, respectively). AA with constant content in the tested albumens were methionine, tryptophan and alanine. The highest content of EAA (>~42 g/100 g of protein) and NAA (>~62 g/100 g of protein) were determined in albumen of eggs determined in the group with at least 20% additions of lupine. The highest content of EAA for humans delivered eggs from groups 4-6 (with the addition of soy into the diet ≤5%). The protein sources used in the hen diet significantly influenced the content of protein and individual AA in the produced eggs.

Salt induced slowdown of kinetics and dynamics during thermal gelation of egg-yolk.

We investigated the effect of the NaCl concentration (0.3-2M) on the structure and dynamics of hen egg yolk at room temperature and during thermal gelation at temperatures in the range of 66-90 °C utilizing low-dose x-ray photon correlation spectroscopy in ultra-small angle x-ray scattering geometry. With an increase in the salt concentration, we observe progressive structural and dynamic changes at room temperature, indicating the disruption of yolk components such as yolk-granules and yolk-plasma proteins. Temperature- and salt-dependent structural and dynamic investigations suggest a delay in the gel formation and aggregation of yolk low-density lipoproteins with increasing ionic strength. However, the time-temperature superposition relationship observed in all samples suggests an identical mechanism underlying protein aggregation-gelation with a temperature-dependent reaction rate. The sol-gel transition time extracted from kinetic and dynamic information follows Arrhenius's behavior, and the activation energy (460kJ/mol) is found to be independent of the salt concentration.

Kaempferia galanga L. alleviates the expression of COX-2 and NF-kappaB-p65 in the oral mucosa ulcer of Wistar rats and exhibits no irritation toward the vascular membrane of chicken eggs and the skin of albino rabbits

The global incidence of recurrent aphthous stomatitis in 2018 reached 5–66 % of the population, while in Indonesia 8 %. Moreover, the prevalence of oral mucosal fibrosis and recurrent aphthous stomatitis among male doctors and nurses in China was 21.24 % and 24.27 %, respectively. Our previous study has shown that the ethanol extract of Kaempferia galanga L. rhizome (EKGR) revealed an accelerated wound-healing effect in the oral mucosa ulcer of Wistar rats. This study aims to explore the effects of EKGR on the expression of NF-kappaB-p65 and COX-2 in the tongue tissue of male Wistar rats by Western blot analysis and immunohistochemistry technique, its safety towards the vascular membrane of the egg chorioallantoic membrane, and its single-dose application on the skin of male rabbits. The rats were randomly assigned into 7 groups: the normal control; the negative control; the positive control (treated with triamcinolone acetonide); and 4 treatment groups of EKGR (0.5 %; 1 %; 2 %; 4 %). Western blot and immunohistochemistry methods were used to measure the expression of NF-kappaB-p65 and COX-2. The hen's egg test-chorioallantoic membrane assay was employed to predict the safety of EKGR towards the vascular membrane. Moreover, the effect of 200 mg/kg BW EKGR application on the dorsal skin of male albino rabbits was also evaluated. EKGR inhibits the expression of NF-kappaB-p65 and COX-2 as proven by WB and IHC results. In the HET-CAM assay, all concentrations of EKGR do not induce irritation responses, which elicits the safety of EKGR. The administration of EKGR causes mild irritation to the dorsal skin of male rabbits but does not induce erythema and edema, no significant changes in BW, no toxic effects on organ macroscopic examination or histopathology, and does not induce abnormalities in the hematological profile of male albino rabbits. EKGR has confirmed its anti-inflammatory activity by suppressing the expression of COX-2 and NF-kappaB-p65 in the oral mucosa ulcer of Wistar rats. EKGR is safe as it does not exhibit irritating potential and harmful effects.

Trajectories of egg sensitization in childhood: Two birth cohorts in Asia and Europe.

Hen's egg exposure through impaired skin barrier is considered a major mechanism of sensitization to eggs. However, the impact of filaggrin (FLG) gene loss-of-function mutations on the natural history of egg sensitization lacks consensus among studies. To evaluate the association between the natural course of egg sensitization and FLG mutations. We used Japanese and the UK birth cohorts (CHIBA and MAAS) to identify the longitudinal patterns of egg sensitization until mid-school age and examined the relationship between the identified patterns and FLG mutations. Sensitization was assessed using egg white-specific IgE levels or skin prick tests (SPTs). Egg allergy was confirmed by parental reports and sensitization. Latent class growth analysis identified longitudinal patterns. Three similar patterns of egg sensitization (persistent, early-onset remitting, and no/low grade classes) were identified in both cohorts, with differing prevalence estimates. The proportion of children with egg allergy in the persistent class at 7 or 8 years of age was 23% (CHIBA) and 20% (MAAS). Consistently in both cohorts, FLG mutations were significantly associated only with the persistent class. Children with FLG mutations had an approximately four-fold increased risk of being in the persistent sensitization class (RRRs: 4.3, 95%C.I. (1.2-16.0), p = .03 in CHIBA; 4.3 (1.3-14.7), p = .02 in MAAS). FLG loss-of-function mutations are associated with persistent egg sensitization in both Japanese and European ethnicities, and the mutations might be a potential biomarker for identifying the risk of persistent egg sensitization/allergy in early infancy. Future studies should incorporate oral food challenges to confirm this relationship.

Effects of black soldier fly (Hermetia illucens) larvae meal on production performance, egg quality, and physiological properties in laying hens: A meta-analysis.

The primary components of fat and protein in chicken diets are fishmeal and soybean; however, due to limited supply and high costs, several efforts have been made to utilize alternative feedstuffs. The potential of black soldier fly larvae (BSFL) as a substitute for fat and protein has been extensively studied, but the findings are not consistent. This study used a meta-analysis approach to investigate the integrated efficacy of BSFL supplementation on laying hen production performance, egg quality, and physiological properties. The articles were retrieved from PubMed, Scopus, ScienceDirect, Cochrane Library, and ProQuest. The retrieved references were examined for potential inclusion. The relevant findings of the included studies were then extracted. Fixed-effects, standard mean difference, 95% confidence intervals, and heterogeneity models were analyzed using the Review Manager website version (Cochrane Collaboration, UK). A total of 24 papers from 17 different nations across five continents have been selected for meta-analysis out of the 3621 articles that were reviewed. The current meta-analysis demonstrated that providing BSFL meals significantly favored feed efficiency, haugh units, albumen quality, eggshell quality, serum glucose, and lipid levels. In addition, significant trends in alanine transaminase, alkaline phosphatase, magnesium, phosphorus, chlorine, and iron levels were observed in blood urea nitrogen, uric acid, creatinine, lactate dehydrogenase, creatine kinase, glutathione peroxidase, and malondialdehyde. On the other hand, it was revealed that there was no favorable effect on weight gain, laying, yolk quality, and hematological profile. The meta-analysis confirmed that BSFL meals can be utilized to optimize feed efficiency, haugh units, albumen, eggshell quality, liver, renal, and cellular physiology of laying hens, although they did not significantly increase body weight gain, laying production, and hematological profiles.

Evaluation of black soldier fly larvae and protease supplementation of Lohmann Brown-lite laying hens performance and egg quality

Abstract Feed costs represent up to 60-70% of the total cost of poultry production, with protein sources being the most expensive feed component. Black soldier fly larvae meal (BSFLM) shows promise as a sustainable protein source for use in animal diets. This research was conducted to determine the effects of including up to 13% defatted BSFLM and a supplementary protease-based enzyme (Concentrase-P; CBS Bio Platforms, Inc., Calgary AB, Canada) on laying hen performance and egg quality. To test this objective, 180 Lohmann Brown-Lite hens (52 weeks of age) were fed experimental diets containing 0%, 6.5%, or 13% BSFLM, with or without Concentrase-P (3 × 2 factorial arrangement of treatments). The treatments were fed over 20 weeks. Results indicated reduced feed intake and body weight in laying hens fed a 13% dietary inclusion level of BSFLM (), while other performance traits (egg production and FCR) were similar among treatments () and met industry performance standards. Shell weight and egg breaking strength did not significantly change with the dietary inclusion level of BSFLM or enzyme inclusion (), although a significant decrease in egg weight and albumen height was noted (), as well as a significant decrease in albumen height for the Concentrase-P treatments (). Yolk colours were significantly lighter and redder in birds fed diets containing high levels of BSFLM (). Results support the use of BSFLM at dietary inclusion levels up to 13% in laying hen diets. Concentrase-P does not appear to be required in combination with BSFLM in terms of growth and laying performance, although potential benefits to other physiological factors are worth consideration.

Insulin-Mimetic Activity of Herbal Extracts Identified with Large-Scale Total Internal Reflection Fluorescence Microscopy.

Diabetes mellitus is a spreading global pandemic. Type 2 diabetes mellitus (T2DM) is the predominant form of diabetes, in which a reduction in blood glucose uptake is caused by impaired glucose transporter 4 (GLUT4) translocation to the plasma membrane in adipose and muscle cells. Antihyperglycemic drugs play a pivotal role in ameliorating diabetes symptoms but often are associated with side effects. Hence, novel antidiabetic compounds and nutraceutical candidates are urgently needed. Phytogenic therapy can support the prevention and amelioration of impaired glucose homeostasis. Using total internal reflection fluorescence microscopy (TIRFM), 772 plant extracts of an open-access plant extract library were screened for their GLUT4 translocation activation potential, resulting in 9% positive hits. Based on commercial interest and TIRFM assay-based GLUT4 translocation activation, some of these extracts were selected, and their blood glucose-reducing effects in ovo were investigated using a modified hen's egg test (Gluc-HET). To identify the active plant part, some of the available candidate plants were prepared in-house from blossoms, leaves, stems, or roots and tested. Acacia catechu (catechu), Pulmonaria officinalis (lungwort), Mentha spicata (spearmint), and Saponaria officinalis (common soapwort) revealed their potentials as antidiabetic nutraceuticals, with common soapwort containing GLUT4 translocation-activating saponarin.

Engineered Lysozyme: An Eco‐Friendly Bio‐Mechanical Energy Harvester

Eco‐friendly and antimicrobial globular protein lysozyme is widely produced for several commercial applications. Interestingly, it can also be able to convert mechanical and thermal energy into electricity due to its piezo‐ and pyroelectric nature. Here, we demonstrate engineering of lysozyme into piezoelectric devices that can exploit the potential of lysozyme as environmentally friendly, biocompatible material for mechanical energy harvesting and sensorics, especially in micropowered electronic applications. Noteworthy that this flexible, shape adaptive devices made of crystalline lysozyme obtained from hen egg white exhibited a longitudinal piezoelectric charge coefficient (d ~ 2.7 pC N−1) and piezoelectric voltage coefficient (g ~ 76.24 mV m N−1) which are comparable to those of quartz (~2.3 pC N−1 and 50 mV m N−1). Simple finger tapping on bio‐organic energy harvester (BEH) made of lysozyme produced up to 350 mV peak‐to‐peak voltage, and a maximum instantaneous power output of 2.2 nW cm−2. We also demonstrated that the BEH could be used for self‐powered motion sensing for real‐time monitoring of different body functions. These results pave the way toward self‐powered, autonomous, environmental‐friendly bio‐organic devices for flexible energy harvesting, storage, and in wearable healthcare monitoring.

Accidental allergic reactions to immediate-type food allergens in Japanese children: A single-center study.

Few studies have assessed the nature of accidental allergic reactions (AAR). We assessed the prevalence and risk factors for AAR in Japanese children. This study included children with immediate-type hen's egg (HE), cow's milk (CM), wheat, or peanut allergy who developed allergic reactions within at least 2 years and were followed up regularly at a single national allergy center in Japan. From January to December 2020, low-dose reactivity was defined as allergic reactions to ≤250, ≤102, ≤53, or ≤ 133 mg of HE, CM, wheat, or peanut protein, respectively. The annualized AAR rate showed the number of reactions per patient per year (95% confidence interval). AAR risk factors were analyzed using multiple logistic regression. Of the 1096 participants, 609, 457, 138, and 90 had HE, CM, wheat, and peanut allergies, respectively. The median (interquartile range) age was 5.0 (2.3-8.6) years, 39% had completely eliminated allergenic food, and 24% had low-dose reactivity. The annualized AAR rate was 0.130 (0.109-0.153) in all sub-cohorts. Moderate and severe symptoms occurred in 50% and 0.7%, respectively, of children who experienced AAR. Multiple logistic regression revealed that low-dose reactivity was a significant risk factor for AAR in the overall and CM cohorts, respectively (p < .001 and p = .036). In this single-center study in Japan, the annualized AAR rate was relatively low during the COVID-19 pandemic; however, half of the participants with AAR had moderate to severe symptoms. Especially in the case of low-dose reactivity, children would require careful AAR risk management.

Screening for emerging chemical risks in the food chain (SCREENER)

Abstract In the European Union, all chemical substances that are manufactured, imported or used in tonnages equal or higher than one tonne per year must be registered under the REACH regulation. This regulatory framework requires the collection and dissemination of detailed information on the properties, uses, and potential health and environmental impacts of chemicals. However, some of these chemicals may pose emerging risks in the food chain, necessitating the development of approaches to identify and assess such risks. The SCREENER project addressed this critical issue by analysing food items for the presence of chemicals that may pose unrecognised hazards. Building on the findings from the previous REACH 1 and REACH 2 projects, which identified a suspect list of 212 chemicals, qualitative multi‐residue high‐resolution mass spectrometry (HRMS) methods have been developed. Diverse food items such as wheat flour, kale, carrots, potatoes, peas, strawberries, oranges, meat, hen's eggs, and trout were pooled from 3 samples each, while cow's milk, and other fish types (salmon and herring) were pooled from 4 samples each to form a single sample. This pooling procedure resulted in 194 analytical samples, which were screened for the presence of the chemicals in the suspect list. Additionally, non‐target analysis (NTA) was performed on the same samples, focusing on detecting halogenated compounds. The aim was to detect substances potentially introduced into the food chain unintentionally through industrial and anthropogenic activities, which were previously unrecognised in the food chain. In the final stage of the project, 15 chemicals were further prioritized for identification and quantitative analysis. Quantitative methods were developed and then applied to the same 194 samples, allowing for the identification and quantification of those chemicals. Subsequently, hazard characterization, exposure assessment, and risk characterisation were conducted. A preliminary characterization of the potential risk posed by the chemicals found in the samples during quantitative confirmatory analysis indicated no risk to human health in all but three cases where additional analysis on occurrence and detailed evaluation of the hazard can be considered as a possible follow‐up action in the future.

The risk of acquiring avian influenza from commercial poultry products and hen eggs: A qualitative assessment

High pathogenicity and low pathogenicity avian influenza (HPAI and LPAI) viruses primarily infect birds, but they can also cause illness in other species, including humans. Some avian influenza (AI) strains can cause fatality rates of over 50 % in human infections. In October 2021, there was a substantial increase in the number of AI infections reported in birds in the UK. Given concerns that more infected and/or contaminated poultry products might reach retail, a risk assessment was performed to ensure that advice relating to the handling and consumption of these products remained appropriate.The products considered in this risk assessment were commercial chicken and turkey products, farmed duck and geese products, and table eggs. The risk pathway included the likelihood animals or eggs from an infected flock would be sent for further processing, whether the resulting products would be released to retail after inspection, viral persistence during distribution and storage, and the ability of AI viruses to infect humans via the gastrointestinal route. The risk from any AI virus, not just the A(H5N1) strain that began circulating in 2021, was considered. Data was obtained from literature searches and FSA surveys.The risk assessment determined that the likelihood of human infection with AI from poultry products for the UK population from handling and consuming commercial chicken or turkey products was negligible with low uncertainty, and for farmed duck and geese products was very low with medium uncertainty. The likelihood of infection for people in the UK from handling and consuming hen table eggs was very low with low uncertainty. The uncertainty rankings relate to the differing amounts of data available for each group of poultry products. The severity of illness in humans from AI infection was considered high with medium uncertainty. The conclusions of this risk assessment for UK consumers largely reflected advice and assessments from other countries and previous UK assessments. Given this, current guidance for handling and consuming poultry products was considered appropriate despite the increase in infections in birds during the 2021/22 and 2022/23 avian flu seasons. Since AI viruses were considered generally, these risk characterisations may need to be revisited based on evidence specific to a circulating virus to support risk management decisions.

Physical Quality Parameters and Activity of Albumen Lysozyme of Eggs From Traditional Hens Bred in Poland Compared With Commercial Hybrids

Abstract The variation in egg quality parameters of traditional/native hen breeds with good freshness and antibacterial properties of egg albumen may determine the attractiveness of these eggs for modern consumers looking for high quality products. The objective of this study was to evaluate external and internal physical quality parameters and enzymatic activity of lysozyme of eggs from three traditional hens bred in Poland and to compare these egg quality parameters to those obtained from commercial hybrids kept under the same management conditions in extensive farming production system. The study was carried out on eggs collected from 4 genetic groups of hens, i.e. Polish Liliputy Bantams (PLB), native Polish Crested Chickens (PCr, CP-22 strain), Gold Laced Polish Chickens (GLP) and from Hy–Line Brown hybrids (HLB). In total 135 PLB, 75 PCr, 75 GLP and 75 HLB hens were kept on litter (3 pens) and fed commercial feed. The eggs (n = 33/genotype/age) were collected at the 33 and 55 weeks of age. The study concluded that with the age of the hens, there was an increase (P&lt;0.05) in the weight of the egg and the proportion of its main fractions, as well as a decrease (P&lt;0.05) in the quality parameters of the albumen and the eggshell. However the eggs from traditional breeds retain good parameters of albumen quality even at the end of production period. The eggs of traditional hens were characterized by higher (P&lt;0.05) yolk proportion and its color and albumen lysozyme parameters, and a lower (P&lt;0.05) eggshell and albumen height and Haugh unit score in relation to commercial hybrids. The highest (P&lt;0.05) content of lysozyme and enzymatic activity of lysozyme were found in eggs from Gold Laced Polish Chickens. Thus, eggs obtained from traditional hens may meet requirements of modern consumers because of specific physical characteristics, good albumen lysozyme activity and freshness parameters. Particularly noteworthy are Polish Liliputy Bantams eggs, characterized by the low weight and a high proportion of more intensely colored yolk and good Haugh unit score at the end of production period.

The volatile constituents of rose, saffron and cardamom suppress the fibrillation of tau and not HEWL through the formation of non-toxic tau oligomers

The formation of fibrillar structures of tau is correlated with taupathies including Alzheimer's disease (AD). This study has aimed to find ways to prevent tau fibril formation. Here, we used dietary compounds including cinnamon (CN), damask rose (Rose), saffron (Saf) and green cardamom (Car), to evaluate the effects of their volatile constituents, on hen egg white lysozyme (HEWL), as a model protein (commonly used for fibrillation studies), as well as the brain-related tau protein. The study was done using different spectroscopic techniques as well as SDS-PAGE, AFM and MTT assay. While the results suggested that the volatile constituents were unable to prevent HEWL fibril formation, most of the dietary compounds, in particular Saf, Rose and Car, were able to interfere with the mature fibril formation, by either maintaining the native form of tau or resulting in the entrapment of non-toxic oligomeric forms of tau. Moreover, the neurotoxicity analysis of tau samples on neuroblastoma SH-SY5Y cells indicated that tau treated with Saf, Rose and Car were the least toxic. Overall, the findings indicate that the potential therapeutic impacts of the volatile constituents of Rose, Car and in particular Saf, may demonstrate significant efficacy in addressing neurodegenerative diseases such as AD.

Real-time observation of a metal complex-driven reaction intermediate using a porous protein crystal and serial femtosecond crystallography

Determining short-lived intermediate structures in chemical reactions is challenging. Although ultrafast spectroscopic methods can detect the formation of transient intermediates, real-space structures cannot be determined directly from such studies. Time-resolved serial femtosecond crystallography (TR-SFX) has recently proven to be a powerful method for capturing molecular changes in proteins on femtosecond timescales. However, the methodology has been mostly applied to natural proteins/enzymes and limited to reactions promoted by synthetic molecules due to structure determination challenges. This work demonstrates the applicability of TR-SFX for investigations of chemical reaction mechanisms of synthetic metal complexes. We fix a light-induced CO-releasing Mn(CO)3 reaction center in porous hen egg white lysozyme (HEWL) microcrystals. By controlling light exposure and time, we capture the real-time formation of Mn-carbonyl intermediates during the CO release reaction. The asymmetric protein environment is found to influence the order of CO release. The experimentally-observed reaction path agrees with quantum mechanical calculations. Therefore, our demonstration offers a new approach to visualize atomic-level reactions of small molecules using TR-SFX with real-space structure determination. This advance holds the potential to facilitate design of artificial metalloenzymes with precise mechanisms, empowering design, control and development of innovative reactions.

Effect of Bile Acids Supplementation in Fatty Liver Hemorrhagic Syndrome, Production Performance, Physiological and Quality Characteristics of Laying Hen Eggs.

This study aimed to investigate the effects of bile acids (BAs) supplementation on fatty liver hemorrhagic syndrome (FLHS), production performance, and physiological and quality characteristics of laying hen eggs. Sixty Sanhuang laying hens, aged 28 weeks, were randomly allocated to six dietary treatments over a 4-week period, including the control (CON) group (feeding basal diet), the high-fat diet (HFD)-treated group (basal diet containing 10% soybean oil), and HFD supplemented with 0.01% and 0.02% of chenodeoxycholic acid (CDCA) or hyodeoxycholic acid (HDCA) groups. Production performance, egg quality, liver morphology, serum biochemical indexes, antioxidant capacity, proinflammatory cytokines, and intestinal microbiota were evaluated. The average body weight in 0.01% CDCA was larger than in the HFD group (p < 0.05). Eggshell Thickness in the CON group was greater than in the HFD, 0.01% CDCA, and HDCA groups (p < 0.05). Albumen height in the 0.02% HDCA group was higher than the HFD group (p < 0.05). Eggshell weight in the HFD group was less than the CON group (p < 0.05). Haugh unit (HU) in the HDCA group was larger than the HFD group (p < 0.05). Albumen weight in the 0.02% HDCA group was greater than the CON and HFD groups (p < 0.05). In the HFD group, the levels of triglyceride (TG), total cholesterol (TC), and low-density lipo-protein cholesterol (LDL-C) were surpassing the other groups (p < 0.05). The levels of catalase (CAT) and total superoxide dismutase (T-SOD) in the HFD group was smaller than the other groups (p < 0.05). The level of malondialdehyde (MDA) in the HFD group was higher than in the other groups (p < 0.05). Tumor necrosis factor-α (TNF-α) levels were larger in the HFD group than in the other groups (p < 0.05). The 16S rRNA sequencing analysis indicated significant variations in the relative abundance of specific bacterial populations among the different treatment groups. The treatment and CON groups exhibited a higher presence of bacteria that inhibit host energy absorption or promote intestinal health such as Firmicutes, Bacteroidetes, and Ruminococcus, whereas the HFD group showed an increased prevalence of potentially pathogenic or deleterious bacteria, such as Desulfovibrio spp. In conclusion, the supplementation of BAs in poultry feed has been demonstrated to effectively mitigate the detrimental effects of FLHS in laying hens. This intervention regulates lipid metabolism, bolsters antioxidant defenses, reduces inflammation, and modulates the gut microbiota, offering a novel perspective on the application of BAs in the poultry industry.

Antioxidant Extracts from Greek and Spanish Olive Leaves: Antimicrobial, Anticancer and Antiangiogenic Effects.

Olea europaea L. is the most valuable species of the Olea type, and its products offer a wide range of therapeutical uses. The olive tree has been extensively studied for its nourishing qualities, and the "Mediterranean diet", which includes virgin olive oil as a key dietary component, is strongly associated with a reduced risk of cardiovascular disease and various malignancies. Olive leaves, a by-product in the olive harvesting process, are valued as a resource for developing novel phytomedicines. For this purpose, two ethanolic extracts obtained from Olivae folium from Spain (OFS) and Greece (OFG) were investigated. Our findings contribute to a wider characterization of olive leaves. Both extracts displayed important amounts of phenolic compounds and pentacyclic triterpenes, OFG having higher concentrations of both polyphenols, such as oleuropein and lutein, as well as triterpenes, such as oleanolic acid and maslinic acid. The antioxidant capacity is similar for the two extracts, albeit slightly higher for OFG, possibly due to metal polyphenol complexes with antioxidant activity. The extracts elicited an antimicrobial effect at higher doses, especially against Gram-positive bacteria, such as Streptococcus pyogenes. The extract with lower inorganic content and higher content of polyphenols and triterpenic acids induced a strong anti-radical capacity, a selective cytotoxic effect, as well as antimigratory potential on A375 melanoma cells and antiangiogenic potential on the CAM. No irritability and a good tolerability were noted after evaluating the extracts on the in vivo Hen's Egg Test-Chorioallantoic Membrane (HET-CAM). Therefore, the present data are suggestive for the possible use of the two types of olive leaf products as high-antioxidant extracts, potentially impacting the healthcare system through their use as antimicrobial agents and as anticancer and anti-invasion treatments for melanoma.

A comprehensive in-vitro/in-vivo screening toolbox for the elucidation of glucose homeostasis modulating properties of plant extracts (from roots) and its bioactives.

Plant extracts are increasingly recognized for their potential in modulating (postprandial) blood glucose levels. In this context, root extracts are of particular interest due to their high concentrations and often unique spectrum of plant bioactives. To identify new plant species with potential glucose-lowering activity, simple and robust methodologies are often required. For this narrative review, literature was sourced from scientific databases (primarily PubMed) in the period from June 2022 to January 2024. The regulatory targets of glucose homeostasis that could be modulated by bioactive plant compounds were used as search terms, either alone or in combination with the keyword "root extract". As a result, we present a comprehensive methodological toolbox for studying the glucose homeostasis modulating properties of plant extracts and its constituents. The described assays encompass in-vitro investigations involving enzyme inhibition (α-amylase, α-glucosidase, dipeptidyl peptidase 4), assessment of sodium-dependent glucose transporter 1 activity, and evaluation of glucose transporter 4 translocation. Furthermore, we describe a patch-clamp technique to assess the impact of extracts on KATP channels. While validating in-vitro findings in living organisms is imperative, we introduce two screenable in-vivo models (the hen's egg test and Drosophila melanogaster). Given that evaluation of the bioactivity of plant extracts in rodents and humans represents the current gold standard, we include approaches addressing this aspect. In summary, this review offers a systematic guide for screening plant extracts regarding their influence on key regulatory elements of glucose homeostasis, culminating in the assessment of their potential efficacy in-vivo. Moreover, application of the presented toolbox might contribute to further close the knowledge gap on the precise mechanisms of action of plant-derived compounds.

Reactive Species Risk Assessment Using Optimized HET-CAM Safety Evaluation of Feed Gas-Modified Gas Plasma Technology and Anticancer Drugs.

Clinical therapies, including dermatology and oncology, require safe application. In vitro experiments allow only limited conclusions about in vivo effects, while animal studies in, e.g., rodents have ethical constraints at a large scale. Chicken embryos lack pain reception until day 15 postfertilization, making the in ovo model a suitable alternative to in vivo safety assessment. In addition, the hen's egg test on chorioallantoic membrane assay allows irritation potential analysis for topical treatments, but standardized analysis has been limited so far. Medical gas plasma is a topical, routine, approved dermatology treatment. Recent work suggests the potential of this technology in oncology. Its main mode of action is the release of various reactive species simultaneously. Intriguingly, varying plasma feed gas compositions generates customized reactive species profiles previously shown to be optimized for specific applications, such as skin cancer treatment. To support clinical implications, we developed a novel chicken embryo CAM scoring and study scheme and employed the model to analyze 16 different plasma feed gas settings generated by the atmospheric pressure plasmajet kINPen, along with common anticancer drugs (e.g., cisplatin) and physiological mediators (e.g., VEGF). Extensive gas- and liquid-phase plasma reactive species profiling was done and was found to have a surprisingly low correlation with irritation potential parameters. Despite markedly different reactive species patterns, feed gas-modulated kINPen plasma was equally tolerated compared to standard argon plasma. CAM irritation with gas plasmas but not anticancer agents was reversed 48 h after treatment, underlining the only temporary tissue effects of medical gas plasma. Our results indicate a safe therapeutic application of reactive species.