Whey Acidic Protein Research Articles

Regulation on Aggregation Behavior and In Vitro Digestibility of Phytic Acid-Whey Protein Isolate Complexes: Effects of Heating, pH and Phytic Acid Levels.

The impact of heat treatment, pH and phytic acid (PA) concentration on the aggregation behavior and digestibility of whey protein isolate (WPI) was investigated. The experimental results indicated that below the isoelectric point of WPI, heat treatment and elevated PA levels significantly increased turbidity and particle size, leading to the aggregation of WPI molecules. No new chemical bonds were formed and the thermodynamic parameters ΔH < 0, ΔS > 0 and ΔG < 0 suggested that the interaction between PA and WPI was primarily a spontaneous electrostatic interaction driven by enthalpy. After the small intestine stage, increasing phytic acid levels resulted in a significant decrease in hydrolysis degree from 16.2 ± 1.5% (PA0) to 10.9 ± 1.4% (0.5% PA). Conversely, above isoelectric point of WPI, there was no significant correlation between the presence of PA and the aggregation behavior or digestion characteristics of WPI. These results were attributed to steric hindrance caused by PA-WPI condensates, which prevented protease binding to hydrolysis sites on WPI. In summary, the effect of PA on protein aggregation behavior and digestive characteristics was not simply dependent on its presence but largely on the aggregation degree of PA-WPI induced by heat treatment, pH and PA concentration. The findings obtained here suggested that phytic acid may be utilized as an agent to modulate the digestion characteristics of proteins according to production requirements. Additionally, the agglomerates formed by heating phytic acid and protein below the isoelectric point could also be utilized for nutrient delivery.

Versatile whey acidic protein four-disulfide core domain proteins: biology and role in diseases.

The Whey acidic protein four-disulfide core (WFDC) protein family consists of proteins with one or more WFDC domains which are ubiquitously expressed throughout the body of human and perform a wide range of functions, including antiprotease, antibacterial, and immunomodulatory functions. Aberrant expression of WFDC proteins is associated with human diseases. However, review on the WFDC protein family is limited and insufficient. Furthermore, a systematic summary of the underlying mechanisms of WFDC protein activity is lacking. In this review, we give a summary of the structural basis and molecular function of these proteins and review the immune regulatory mechanisms and signaling pathways of WFDC proteins in the development of certain diseases. Furthermore, we discuss the diagnostic and prognostic potential of multiple WFDC proteins in the aforementioned conditions, as well as their prospective use. At last, we also discuss the progress of WFDC protein in clinical trials and put forward some research difficulties and the directions of follow-up research. Our review highlights the functional diversity and clinical significance of WFDC proteins family, while providing potential targets for drug development and innovative therapeutic strategies, this review lays the foundation and direction for future research on WFDC proteins.

Enhancing the water dispersibility and intestinal targeting of pterostilbene using tannic acid-whey protein conjugates

In this study, three types of whey protein isolate (WPI)-tannic acid (TA) covalent conjugates (WPI-TA) were synthesized via alkaline treatment, free radical-induced method, and laccase induction. These conjugates were subsequently utilized in the antisolvent precipitation method to encapsulate pterostilbene (Pte). The structural and functional properties of these conjugates as carriers for intestinal-targeted Pte delivery were thoroughly evaluated. SDS-PAGE, total phenolic content measurement, grafting efficiency, and FT-IR analysis confirmed the successful formation of three distinct covalent conjugates. Among them, the laccase-induced WPI-TA conjugates exhibited the highest TA grafting efficiency. The particle size of this conjugate was measured at 115.25 nm with a PDI of 0.31. Upon encapsulation of Pte, this conjugate exhibited superior thermal stability, enhanced antioxidant properties, and sustained release within the gastrointestinal tract. The water solubility of Pte in these nanoparticles was found to be 361.19 times higher than that of free Pte, presenting a promising strategy for targeted intestinal delivery of Pte.

A sex-specific homologue of waprin is essential for embryonic development in the red flour beetle, Tribolium castaneum.

Waprin, a WAP (Whey acidic protein) domain-containing extracellular secretory protein, is widely known for its antibacterial properties. In this study, a waprin homologue (Tc_wapF) expressing in a female-specific manner was identified in Tribolium castaneum, through the analysis of sex-specific transcriptomes. Developmental- and tissue-specific profiling revealed the widespread expression of Tc_wapF in adult female tissues, particularly in the ovary, gut and fatbody. This female-specific expression of Tc_wapF is not regulated by the classical sex-determination cascade of T. castaneum, as we fail to get any attenuation in Tc_wapF transcript levels in Tcdsx and Tctra (key players of sex determination cascade of T. castaneum) knockdown females. RNA interference-mediated knockdown of Tc_wapF in females led to the non-hatching of eggs laid by these females, suggesting the crucial role of Tc_wapF in the embryonic development in T. castaneum. This is the first report on the identification of a sex-specific waprin homologue in an insect and its involvement in embryonic development. Future investigations on the functional conservation of insect waprins and their mechanistic role in embryonic development can be exploited for improving pest management strategies.

Effect of ultrafiltration of acid whey at pH 4.6 or 6.5 on whey concentrate composition and resultant powder properties

This study investigated the effect of modifying the mineral composition through adjustment of pH prior to ultrafiltration on the functionality of acid whey protein concentrate (WPC; pH 4.6). Increasing pH to 6.5 prior to ultrafiltration produced a concentrate with a significantly higher calcium and phosphorous content, compared to ultrafiltration at pH 4.6. WPC produced at pH 6.5 also had significantly more calcium‐phosphate precipitation following pasteurisation and evaporation and had a higher viscosity at 30% solids (300 s−1, 50°C). In addition, both reconstituted powders (5%, w/w, protein) had strong gelation properties following heat treatment at 90°C; however, the WPC produced at pH 6.5 had a significantly firmer gel strength, but such gels appeared to be more sensitive to breakage at increased frequency and oscillation strain. Overall, the pH prior to UF had a significant impact on the processing of acid whey and final product functionality.

The Study of Combination of Biodegradable Packaging and Biocoating with Lactic Acid Bacteria as a Green Alternative for Traditional Packaging in Gouda Cheese

Biodegradable packaging, both alone and in combination with acid whey protein coatings, has been used to pack fresh Gouda cheese to improve preservation prior ripening or storage. This study involved three key components: (i) the selection of biodegradable packaging (BP), (ii) the development of a plain liquid acid whey protein concentrate, pectin-based edible coating (BP + Ch + Coating), and (iii) the incorporation of at least 6 log10 CFU (colony forming units) mL−1 Lacticaseibacillus paracasei (BP + Ch + Coating + Lp) and Lactobacillus helveticus (BP + Ch + Coating + Lh) strains. The created compositions were compared with cheese packed in conventional polyethylene (PE) packaging to evaluate their overall synergy effect in reducing microbiological spoilage and influencing chemical parameters in Gouda cheese during 45 days of ripening and cold storage. The evaluation included microbiological analysis (total LAB, Enterobacteriaceae spp., and fungi CFU) and quality assessment of pH, moisture content, water activity, texture, and colour (CEI system) during ripening and shelf life. Although biodegradable packaging (BP) alone did not protect the cheese effectively compared to conventional packaging (EVA/PE/EPC/PVDC), the combination of biodegradable packaging with a coating (BP + Ch + Coating) showed protective properties against Enterobacteriaceae spp. and mould, maintaining moisture, pH, and colour during ripening and storage. Incorporation of L. helveticus (BP + Ch + Coating + Lh) into the coating efficiently decreased the growth of fungi.

The shell formation mechanism of Turbo argyrostomus based on ultrastructure and transcriptome analysis

The gold inner shell of Turbo argyrostomus is an important morphological classification characteristic in Gastropoda. However, the gene sets responsible for shell formation in gastropods remain poorly explored. In this study, we investigated the microstructure using scanning electron microscopy (SEM), hematoxylin-eosin (HE) and Alcian blue staining-periodic acid-Schiff (AB-PAS) staining. The SEM results illustrated that the T. argyrostomus shell exhibited a special “sandwich” microstructure. The results of histological observation demonstrated two major cell types: adipocytes and mucin cells. A total of 318 differentially expressed genes were identified between edge mantle and central mantle, among which whey acidic protein, N66, and nacre-like proteins, and Lam G and EGF domains may be related to shell microstructure. 22.39% − 25.20% of the mucin genes had biomineralization related domains, which supported for the relationship between mucins and shell formation. Moreover, this study revealed energy distribution differences between the edge mantle and central mantle. These results provide insights for further understanding of the biomineralization mechanism in Gastropoda.

WFDC2 is a potential prognostic and immunotherapy biomarker in lung adenocarcinoma.

The prognosis of lung adenocarcinoma (LUAD), which is the most common type of lung cancer, remains poor. Little is known about the function and mechanism of whey acidic protein four-disulfide core domain 2 (WFDC2) in LUAD. In this study, we used online databases to compare WFDC2 expression between LUAD and normal tissues, to analyze the relationship between WFDC2 and overall survival, and to investigate the potential roles of WFDC2. We found that WFDC2 protein and mRNA expression levels were significantly higher in LUAD tissue than in normal tissue, and high WFDC2 mRNA expression was associated with better overall survival. WFDC2 mRNA expression was correlated with the mutation status of TP53. The biological function of WFDC2 was associated with the cell cycle, and low WFDC2 mRNA expression was associated with an elevated tumor mutational burden and neoantigen levels. A negative relationship was observed between WFDC2 and immune gene expression, and high WFDC2 mRNA expression was found in patients with LUAD and low programed cell death 1 mRNA expression. We propose that WFDC2 is associated with clinical benefits of immune checkpoint inhibitors in LUAD.

Adipo-Epithelial Transdifferentiation in In Vitro Models of the Mammary Gland.

Subcutaneous adipocytes are crucial for mammary gland epithelial development during pregnancy. Our and others' previous data have suggested that adipo-epithelial transdifferentiation could play a key role in the mammary gland alveolar development. In this study, we tested whether adipo-epithelial transdifferentiation occurs in vitro. Data show that, under appropriate co-culture conditions with mammary epithelial organoids (MEOs), mature adipocytes lose their phenotype and acquire an epithelial one. Interestingly, even in the absence of MEOs, extracellular matrix and diffusible growth factors are able to promote adipo-epithelial transdifferentiation. Gene and protein expression studies indicate that transdifferentiating adipocytes exhibit some characteristics of milk-secreting alveolar glands, including significantly higher expression of milk proteins such as whey acidic protein and β-casein. Similar data were also obtained in cultured human multipotent adipose-derived stem cell adipocytes. A miRNA sequencing experiment on the supernatant highlighted mir200c, which has a well-established role in the mesenchymal-epithelial transition, as a potential player in this phenomenon. Collectively, our data show that adipo-epithelial transdifferentiation can be reproduced in in vitro models where this phenomenon can be investigated at the molecular level.

Effect of high-pressure homogenization on maize starch-stearic acid and maize starch-stearic acid-whey protein complexes

The maize starch (MS)-stearic acid (SA) and MS-SA-whey protein (WP) complexes were prepared using the high-pressure homogenization (HPH). Results from X-ray diffraction (XRD) showed that MS-SA complexes presented an increase in the long-range molecular order with increasing the homogenization pressure, and MS-SA-WP complexes showed only an increase with increasing the homogenization pressure from 10 to 50 MPa. Results from differential scanning calorimetry (DSC) and Raman spectroscopy showed that the amount of complexes and the short-range order of both MS-SA and MS-SA-WP complexes increased with increasing the homogenization pressure. The addition of WP to MS-SA altered significantly the structure and digestion of complexes. Results revealed that MS-SA-WP complexes have more ordered structure and lower digestion than the corresponding MS-SA complexes. The digestibility of all complexes decreased with increasing the homogenization pressure. There was a significant correlation between the digestibility and structural characteristics of complexes. Complexes with better structural stability have better anti-digestion properties. The obtained results are helpful in understanding the structure and digestibility of complexes prepared by HPH, which is valuable for controlling the quality and nutrition of starchy food.

Development of Fortified Biscuit with Acid and Sweet Whey Proteins and Its Effects on Body Weight, Indices of Body Composition and Antioxidant Capacity in Rats

The effect of sweet (SW) and acid whey (AW) proteins on body weight (BW) and composition (BC), and antioxidant capacity (AC) is uncertain. We dehydrated SW and AW proteins obtained from local white cheese and labneh production lines, developed biscuits containing these protein powders and evaluated biscuits' sensory attributes and proximate nutrient contents, and investigated their effects on BW, indices of BC, and AC in rats. Four types of biscuit were prepared: plain (PB), albumin (ALB), SWB, and AWB. Five isocaloric and isonitrogenous diets containing Albumin (control), PB, ALB, SWB, and AWB were used. Sixty adult male Sprague-Dawley rats were used, 10 were sacrificed at the start, and the remainder were randomly assigned to the five diet groups (10 rats/ group) and fed for four weeks. BW, food (FI) and water (WI) intake, food efficiency ratio (FER), serum AC, liver weight (LW), Lee index (LI), body mass index (BMI), length, and abdomen (AS) and chest (CC) circumferences were measured following standard protocols. AW was higher in ash and fat whereas lower in carbohydrates but similar to SW in protein and moisture contents. ALB had the highest protein content, AWB had the most increased fat and ash contents, and SWB had the highest moisture content. PB had the highest carbohydrates and the lowest ash contents among the other biscuits. PB and SWB were the best in all sensory attributes tested and the overall acceptability compared to other biscuits. The four biscuit diets kept similar BW, whereas the control diet kept the highest BMI, LI, and LW, which was also not significantly different from the initial group. Animals fed the PB diet had the most increased length, abdomen, AS, and CC, whereas these measurements were lowest in animals fed SWB and AWB diets which were not different from the control. Animals fed the AWB diet had the highest WI, but all other diets were not significantly different from the control diet in FI and FER. Serum AC was the highest in animals fed SWB and AWB diets that were insignificantly different from the initial group, whereas it was the lowest in PB and AWB diets. Given their sensory attributes and effects on AC and BW status, SW and AW may be used as protein substitutes in bakery products; however, further studies are warranted.&#x0D; Keywords: Body weight, Sweet whey, Acidic whey, Proximate nutrients, Body composition, Antioxidant capacity, Rat.

The effect of whey source on heat-induced aggregation of casein and whey protein mixtures of relevance to infant nutritional product formulation

Sweet and, to a lesser extent, acid whey protein ingredients can be used for the formulation of infant nutritional products. Unlike acid whey, sweet whey contains caseinomacropeptide (CMP), a heat-stable peptide liberated from κ-casein during cheese and rennet casein manufacture. Four protein systems-sweet whey (SW) and acid whey (AW), with or without standardization for CMP protein content-were added to skim milk (50/50, wt/wt) and unheated or heated to 85 or 110°C. These 12 samples were assessed for physicochemical stability in the presence of added calcium at pH 6.8. The effect of CMP content on the physicochemical properties of the protein systems was also assessed. Without preheat treatment, mixtures of AW and skim milk (SM) were more heat stable than SW and SM, demonstrating the effect of whey protein type on heat stability. Preheat treatment of the SW in the presence of SM significantly improved the heat stability of the resultant protein systems on subsequent heating. All of the protein systems had significantly lower heat stability with the addition of Ca, although the reduction was significantly smaller for the heated protein systems than the unheated controls. The findings can help identify heating parameters and ingredients for optimizing processing stability and physicochemical characteristics of nutritional beverages such as infant formulations.

High WFDC3 gene expression is associated with poor prognosis and reduced immune cells infiltration in pancreatic adenocarcinoma: A study using the TCGA database and bioinformatics analysis.

Whey-acidic-protein (WAP) four-disulfide core domain protein 3 (WFDC3) is one of the WAP family proteins. This protein family is associated with the development of solid tumors and affects the tumor immunological microenvironment. However, the prognostic value of WFDC3 in pancreatic adenocarcinoma (PAAD) and its effect on the tumor immune microenvironment is yet to be clarified. The Cancer Genome Atlas database and Genotype-Tissue Expression database were used to analyze the differential expression of WFDC3 between the tumor and adjacent tissues. The clinical significance of WFDC3 was analyzed in The Cancer Genome Atlas and International Cancer Genome Consortium database using WFDC3 transcripts and clinical information. In order to elucidate the underlying mechanisms, gene set enrichment analysis was conducted to determine potential activated pathways. Immune score evaluation and publicly available pharmacogenomics database [the Genomics of Drug Sensitivity in Cancer] were utilized to quantify immune cell infiltration and the effect on chemotherapeutic drug sensitivity. WFDC3 levels were higher in PAAD tissues than in normal pancreatic tissues. High levels of WFDC3 expression progressively increased as PAAD tumor stages progressed. Patients with elevated WFDC3 expression showed a poor prognosis. The gene set enrichment analysis analysis revealed that glutamate, arginine, and proline, and histidine metabolism levels were elevated in patients with a high WFDC3 expression phenotype. B, CD4+ T, and CD8+ T cell infiltration was diminished in PAAD tissues with elevated WFDC3 expression. According to pharmacogenomics, PAAD tissues with high WFDC3 expression are susceptible to gemcitabine. WFDC3 is highly expressed in PAAD, and patients with a high level of WFDC3 expression have a shorter overall survival time, indicating a poorer prognosis. High expression of WFDC3 may lead to the development of PAAD by affecting the amino acid metabolism and the tumor immunological microenvironment. WFDC3 may serve as a potential diagnostic and prognostic biomarker for PAAD patients.

Excretory/Secretory Products from Schistosoma japonicum Eggs Alleviate Ovalbumin-Induced Allergic Airway Inflammation.

Excretory/secretory products (ESPs) derived from helminths have been reported to effectively control allergic inflammation, which have better therapeutic prospects than live parasite infections. However, it remains unknown whether ESPs from schistosome eggs can protect against allergies, despite reports alleging that schistosome infection could alleviate disordered allergic inflammation. In the present study, we investigated the protective effects of ESPs from Schistosoma japonicum eggs (ESP-SJE) on asthmatic inflammation. Firstly, we successfully established an allergic airway inflammation model in mice by alum-adjuvanted ovalbumin (OVA) sensitization and challenge. ESP-SJE were administered intraperitoneally on days -1 and 13 (before sensitization), on day 20 (before challenge), and on days 21-24 (challenge phase). The results showed that ESP-SJE treatment significantly reduced the infiltration of inflammatory cells, especially eosinophils into the lung tissue, inhibited the production of the total and OVA-specific IgE during OVA-sensitized and -challenged phases, respectively, and suppressed the secretion of Th2-type inflammatory cytokines (IL-4). Additionally, ESP-SJE treatment significantly upregulated the regulatory T cells (Tregs) in the lung tissue during OVA challenge. Furthermore, using liquid chromatography-mass spectrometry analysis and Treg induction experiments in vitro, we might identify nine potential therapeutic proteins against allergic inflammation in ESP-SJE. The targets of these candidate proteins included glutathione S-transferase, egg protein CP422 precursor, tubulin alpha-2/alpha-4 chain, actin-2, T-complex protein 1 subunit beta, histone H₄, whey acidic protein core region, and molecular chaperone HtpG. Taken together, the results discussed herein demonstrated that ESP-SJE could significantly alleviate OVA-induced asthmatic inflammation in a murine model, which might be mediated by the upregulation of Treg in lung tissues that may be induced by the potential modulatory proteins. Therefore, potential proteins in ESP-SJE might be the best candidates to be tested for therapeutic application of asthma, thus pointing out to a possible new therapy for allergic airway inflammation.

Characterization of a crustin-like peptide involved in shrimp immune response to bacteria and Enterocytozoon hepatopenaei (EHP) infection in Palaemon carinicauda.

Crustins represent one type of antimicrobial peptides (AMPs) that are key components of the innate immune process of crustaceans. This study successfully identified a novel crustin-like peptide, EcCrustin2, in ridgetail white prawn, Palaemon carinicauda (formerly Exopalaemon carinicauda). EcCrustin2 was found to be 1082 bp in length, with a 378 bp open reading frame (ORF) encoding 125 amino acids. The deduced amino acid sequence of EcCrustin2 exhibited characteristics of crustins in crustacean, including a Cys-rich region at the N-terminus as well as a whey acidic protein domain at the C-terminus. Phylogenetic analysis revealed that the EcCrustin2 was first clustered with Type I crustins, then with other crustins. Expression of EcCrustin2 was mainly detected in immune tissues, including hemocytes, gill and stomach. The expression level of EcCrustin2 was also significantly up-regulated after being exposed to lipopolysaccharide (LPS), lipoteichoic acid (LTA), Vibrio parahaemolyticus and Staphylococcus aureus. EHP infection could also induce EcCrustin2 expression in P. carinicauda. Knockdown of EcCrustin2 with siRNA increased the mortality of V. parahaemolyticus challenged shrimp. Finally, the recombinant EcCrustin2 protein was obtained and demonstrated a wide spectrum of antibacterial activity in vitro. These results indicated that EcCrustin2 takes part in the immune response against bacteria and EHP infection.

Cancer stem cells as the source of tumor associated myoepithelial cells in the tumor microenvironment developing ductal carcinoma in situ

The heterogeneous cell population in the stromal microenvironment is considered to be attributed to the multiple sources from which the cells originate. Tumor associated myoepithelial cells (TAMEs) are one of the most important populations in the tumor microenvironment (TME) especially in breast cancer. On the other hand, cancer stem cells (CSCs) have previously been described to be the origin of tumor-associated cellular components in the TME. We prepared a cancer stem cell model converting mouse-induced pluripotent stem cells (miPSCs) in the presence of conditioned medium of breast cancer cell line MDA-MB-231 cells. The converted cells developed tumors progressing into invasive carcinoma with ductal carcinoma in situ (DCIS) like structure when transplanted into mouse mammary fat pads. The primary cultured cells from the tumor further exhibited markers of CSC such as Sox2, Oct3/4, - CD133 and EpCAM, and mammary gland-related TAME markers such as α-smooth muscle actin, cytokeratin 8, whey acidic protein, prolactin receptor and progesterone receptor as well. These results indicated that the CSCs could be an origin of TAMEs contributing to mammary gland epithelial cell differentiation and the progression to invasive carcinoma during tumor development. The gene expression profiles confirmed the enhanced signaling pathways of PI3K/AKT and MAPK, which have been demonstrated to be enriched in the CSC models, together with the estrogen receptor signaling which was peculiar to mammary gland-derived character.

Characterization and immune functional analysis of two new type I crustins in the oriental river prawn Macrobrachium nipponense

Crustins are cationic antimicrobial peptides that are categorized into five types and play a key role in the nonspecific immunity of crustaceans. In the present study, full-length cDNA sequences of two Macrobrachium nipponense crustin genes (MnCrustin1 and MnCrustin2) were cloned and identified. The full-length cDNAs of MnCrustin1 and MnCrustin2 were 715 and 881 bp, including 351 and 330 bp open reading frames (ORFs) encoding 117 and 110 amino acids, respectively. A signal peptide, a cysteine (Cys)-rich region and a whey acidic protein (WAP) domain existed in both MnCrustin1 and MnCrustin2. Phylogenetic analysis and multiple sequence alignment indicated that MnCrustin1 and MnCrustin2 belonged to the type I crustin family. The MnCrustin1 and MnCrustin2 mRNAs were expressed in various tissues of M. nipponense and were highly expressed in hemocytes and gills. Recombinant MnCrustin1 and MnCrustin2 inhibited the growth of both gram-positive and gram-negative bacteria in vitro. After Aeromonas hydrophila and white spot syndrome virus (WSSV) infection, MnCrustin1 and MnCrustin2 expression levels were elevated in the hepatopancreas, hemocytes and gills of M. nipponense. Tests of RNA interference (RNAi) and recombinant protein injection were employed to further study the roles of MnCrustin1 and MnCrustin2 in pathogen infection. The results indicated that silencing MnCrustin1 and MnCrustin2 accelerated the mortality rate of M. nipponense after challenge with A. hydrophila or WSSV, while recombinant MnCrustin1 and MnCrustin2 decreased the mortality rate under these conditions. These results suggest that MnCrustin1 and MnCrustin2 both play prominent roles in the immune response of M. nipponense to bacterial and viral infection and are important immune effectors of this prawn against pathogen infection.

The effects of polylactic acid-whey protein isolated bi-layer film incorporated with ZnO nanoparticles on the quality of common carp Cyprinus carpio

The effects of polylactic acid-whey protein isolated bi-layer film incorporated with ZnO nanoparticles on the quality of common carp Cyprinus carpio

Transcriptome analysis of germ cell changes in male Chinese mitten crabs (Eriocheir sinensis) induced by rhizocephalan parasite, Polyascus gregaria

The parasitism by Polyascus gregaria on Eriocheir sinensis induces feminization of the appearance of male crabs, misleading fishermen to bring them to the breeding ponds as female crabs to cultivate broodstock selection. However, there are few studies on whether P. gregaria feminizes the male germ cells, resulting in a decline in the fecundity of male crabs. Therefore, this study aims to clarify the changes in gene expression levels of male crab testes after being parasitized by P. gregaria through transcriptome sequencing to evaluate the change in fecundity. We selected parasitized and healthy male crabs from a pond culture for comparison of gene expression in germ cells. The results showed that, compared with healthy male crabs, there were 104 genes with significantly different expressions, of which 79 were up-regulated and 25 were down-regulated. These genes are mainly focused on the cytoskeleton pathway in cell components and cellular protein complex assembly in biological processes. Several spermatogenesis-related genes, such as Kazal-type protease inhibitor, which inhibits gelatinolytic activities of sperm proteases, and juvenile hormone esterase 6, which degrades methyl farnesoate, were up-regulated; while the down-regulated expression of certain heat shock proteins may lead to spermatogenic dysfunction. In addition, some immune-related genes, such as double whey acidic protein domain-containing protein and serine proteinase inhibitor 3, were significantly up-regulated. These results indicated that P. gregaria changed the development process and cell structure of male host germ cells to inhibit sperm proliferation and maturation, while multiple immune pathways in the hosts were activated to resist P. gregaria invasion.

WFDC12-overexpressing contributes to the development of atopic dermatitis via accelerating ALOX12/15 metabolism and PAF accumulation

Atopic dermatitis (AD) is a chronic inflammatory skin disease characterized by eczema-like skin lesions, dry skin, severe itching, and recurrent recurrence. The whey acidic protein four-disulfide core domain gene WFDC12 is highly expressed in skin tissue and up-regulated in the skin lesions of AD patients, but its role and relevant mechanism in AD pathogenesis have not been studied yet. In this study, we found that the expression of WFDC12 was closely related to clinical symptoms of AD and the severity of AD-like lesions induced by DNFB in transgenic mice. WFDC12-overexpressing in the epidermis might promote the migration of skin-presenting cells to lymph nodes and increase Th cell infiltration. Meanwhile, the number and ratio of immune cells and mRNA levels of cytokines were significantly upregulated in transgenic mice. In addition, we found that ALOX12/15 gene expression was upregulated in the arachidonic acid metabolism pathway, and the corresponding metabolite accumulation was increased. The activity of epidermal serine hydrolase decreased and the accumulation of platelet-activating factor (PAF) increased in the epidermis of transgenic mice. Collectively, our data demonstrate that WFDC12 may contribute to the exacerbation of AD-like symptoms in DNFB-induced mouse model by enhancing arachidonic acid metabolism and PAF accumulation and that WFDC12 may be a potential therapeutic target for human atopic dermatitis.