Traditional cooking methods decreased the allergenicity of egg proteins.

Egg allergy is one of the most common food allergies globally. This study aimed to assess the impact of four traditional cooking methods on the allergenicity of egg proteins using a comprehensive strategy, including simulated gastrointestinal digestion in vitro, serology experiments, a rat basophilic leukemia (RBL)-2H3 cell degranulation model, and a passive cutaneous anaphylaxis (PCA) mice model, and the structure changes were detected by circular dichroism (CD) spectra and ultraviolet (UV) spectra. The results showed that the processed egg proteins were more readily digested compared to raw egg proteins. The serological experiments revealed a significant reduction in immunoglobulin E binding of egg proteins after thermal treatments (p < 0.05), particularly after frying. Subsequently, the RBL-2H3 cell degranulation experiment demonstrated a marked decrease in the level of egg allergens-induced ß-hexosaminidase release after cooking (p < 0.05). Moreover, the results from the PCA mice model indicated that the increase in vascular permeability was effectively relieved in the treated groups, especially in frying group (p < 0.05). Additionally, the α-helix and ß-turn contents of processed egg proteins were significantly decreased (p < 0.05) compared with native egg proteins. The UV spectra findings showed that all cooking treatments caused significant alterations in the tertiary structure, and fluorescence analysis indicated that cooking decreased the surface hydrophobicity of egg proteins. In conclusion, four traditional cooking methods reduced the allergenicity of egg proteins, particularly frying, and this reduction was associated with structural changes that could contribute to the destruction or masking of epitopes of egg allergens. PRACTICAL APPLICATION: Egg allergy has a serious impact on public health, and there is no ideal treatment method at present. This study demonstrated that four traditional cooking methods (boiling, steaming, baking, and frying) reduced the allergenicity of egg proteins, especially frying, and the results will provide a basis for the development of hypoallergenic egg products.

Anti-allergic effect of vitamin C through inhibiting degranulation and regulating TH 1/TH 2 cell polarization.

BACKGROUND: Food allergy has become a global public health problem. This study aimed to explore the possible anti-allergic effect of vitamin C (VC). A rat basophilic leukemia (RBL)-2H3 cell degranulation model was used to assess the effect of VC on degranulation in vitro, and an ovalbumin (OVA)-induced BALB/c mouse allergy model was used to assess the anti-allergy effect of VC in vivo. RESULTS: In vitro, VC significantly attenuated the release of ß-hexosaminidase, tryptase and histamine, and also reduced cytokine production (interleukins 4 and 6, tumor necrosis factor α) significantly (P < 0.05), with the inhibitory effect demonstrating a positive correlation with VC dose. In vivo, compared with the OVA group, the levels of serum immunoglobulins E and G1 of the VC low-dose (VCL) group (50 mg kg-1 ) and high-dose (VCH) group (200 mg·kg-1 ) were significantly reduced (P < 0.05). Furthermore, the plasma histamine level was also significantly decreased (P < 0.05). Moreover, TH 2 cell polarization in mice of the VCL and VCH groups was significantly inhibited (P < 0.05), promoting the TH 1/TH 2 cell polarization balance. Additionally, VC treatment enhanced the expression of CD80 (P < 0.05) in spleen and small intestine tissues, while significantly inhibiting the expression of CD86 (P < 0.05); notably, high-dose VC treatment was more effective. CONCLUSION: VC exerted an anti-allergic effect through inhibiting degranulation and regulating TH 1/TH 2 cell polarization balance. © 2024 Society of Chemical Industry.

Development and validation a nomogram for predicting new-onset postoperative atrial fibrillation following pulmonary resection.

BACKGROUND: The new-onset postoperative atrial fibrillation (NOPAF) following pulmonary resection is a common clinical concern. The aim of this study was to construct a nomogram to intuitively predict the risk of NOPAF and offered protective treatments. METHODS: Patients who underwent pulmonary resection between January 2018 and December 2020 were consecutively enrolled. Forward stepwise multivariable logistic regression analyses were used to screen independent predictors, and a derived nomogram model was built. The model performance was evaluated in terms of calibration, discrimination and clinical utility and validated with bootstrap resampling. RESULTS: A total of 3583 patients who met the research criteria were recruited for this study. The incidence of NOPAF was 1.507% (54/3583). A nomogram, composed of five independent predictors, namely age, admission heart rate, extent of resection, laterality, percent maximum ventilation volume per minute (%MVV), was constructed. The concordance index (C-index) was 0.811. The nomogram showed substantial discriminative ability, with an area under the receiver operating characteristic curve of 0.811 (95% CI 0.758-0.864). Moreover, the model shows prominent calibration performance and higher net clinical benefits. CONCLUSION: We developed a novel nomogram that can predict the risk of NOPAF following pulmonary resection, which may assist clinicians predict the individual probability of NOPAF and perform available prophylaxis. By using bootstrap resampling for validation, the optimal discrimination and calibration were demonstrated, indicating that the nomogram may have clinical practicality.

Assessment of the effect of glycation on the allergenicity of sesame proteins.

Sesame allergy is a growing concern worldwide. In this study, sesame proteins was glycated with glucose, galactose, lactose and sucrose respectively, and the allergenicity of different glycated sesame proteins were assessed by a comprehensive strategy, including simulated gastrointestinal digestion in vitro, a BALB/c mice model, a rat basophilic leukemia (RBL)-2H3 cell degranulation model and a serological experiment. Firstly, simulated gastrointestinal digestion in vitro showed that glycated sesame proteins were more easily to digest than raw sesame. Subsequently, the allergenicity of sesame proteins was assessed in vivo by detecting the allergic indexes of mice, and results showed that the levels of total immunoglobulin E (IgE) and histamine were reduced in glycated sesame proteins treated mice. Meanwhile, the Th2 cytokines (IL-4, IL-5, and IL-13) were downregulated significantly, demonstrating that sesame allergy was relieved in glycated sesame treated mice. Thirdly, the RBL-2H3 cell degranulation model results showed that the release of ß-hexosaminidase and histamine were decreased to different degrees in glycated sesame proteins treated groups. Notably, the monosaccharide glycated sesame proteins exhibited lower allergenicity both in vivo and in vitro. Furthermore, the study also analyzed the structure alteration of sesame proteins, and the results showed that the secondary structure of glycated sesame proteins were changed (the content of α-helix and ß-sheet were reduced), and the tertiary structure of sesame proteins after glycation modification was also changed (microenvironment around aromatic amino acids was altered). Besides, the surface hydrophobicity of glycated sesame proteins was also reduced except sucrose glycated sesame proteins. In conclusion, this study demonstrated that glycation reduced the allergenicity of sesame proteins effectively, especially glycation with monosaccharides, and the allergenicity reduction might be related to structural changes. The results will provide a new reference for developing hypoallergenic sesame products.

Study on the anti-inflammatory effect of stachyose by inhibiting TLR4/NF-κB signalling pathway in vitro and in vivo.

This study aimed to explore the anti-inflammatory effect of stachyose, a tetrasaccharide extracted from Stachys sieboldii Miq. A lipopolysaccharide (LPS)-stimulated RAW264.7 macrophages model and a dextran sodium sulfate (DSS)-induced ulcerative colitis BALB/C mice model was used to assess the anti-inflammatory effect of stachyose both in vitro and in vivo. The levels of nitric oxide (NO) and cytokines (interleukin-1ß, interleukin-6 and tumour necrosis factor-α) were detected using enzyme-linked immunosorbent assay methods; moreover, haematoxylin-eosin staining was used to observe changes in intestinal morphology of mice. In addition, the possible mechanisms were explored by reverse transcription-polymerase chain reaction and western blot. Results showed that stachyose and four other oligosaccharides (galacto-oligosaccharides, xylo-oligosaccharides, inulin and resistant dextrin) inhibited NO secretion and the production of pro-inflammatory cytokines in LPS-stimulated RAW264.7 macrophages in a dose-dependent manner, whereas stachyose was most effective in vitro. In mice, different doses of stachyose significantly alleviated the symptoms of DSS-induced ulcerative colitis and stachyose also significantly inhibited the production of inflammatory cytokines and myeloperoxidase in vivo. In addition, our findings illustrated that stachyose inhibited expression of toll-like receptor 4 (TLR4) and suppressed the phosphorylation of nuclear factor (NF)-κB p65 both in vitro and in vivo. Taken together, results demonstrated that stachyose exerted anti-inflammatory effect through inhibition of the TLR4/NF-κB signalling pathway.

Sequence analysis of digestion-resistant peptides may be an efficient strategy for studying the linear epitopes of Jug r 1, the major walnut allergen.

Jug r 1, the major allergen of walnut, triggers severe allergic reactions through epitopes. Hence, research on the efficient strategy for analyzing the linear epitopes of Jug r 1 are necessary. In this work, bioinformatics analysis was used to predict the linear epitopes of Jug r 1. Overlapping peptide synthesis was used to map linear epitopes. In vitro simulated gastrointestinal digestion and HPLC-MS/MS were used to identify digestion-resistant peptides. The results showed that six predicted linear epitopes were AA28-35, AA42-49, AA55-62, AA65-73, AA97-104, and AA109-121. AA16-30 and AA125-139 were identified by the sera of walnut allergic patients. Five digestion-resistant peptides were AA19-33, AA40-45, AA54-74, AA96-106, and AA117-137. The predicted results only included one of the linear epitopes identified by sera, while the digestion-resistant peptides covered all. Therefore, the digestion-resistant property of food allergens may be a promising direction for studying the linear epitopes of Jug r 1.

Almond (Prunus dulcis) Allergen Pru du 8, the First Member of a New Family of Food Allergens.

An almond allergen with two known short peptide sequences was reported as the almond 2S albumin but was later suspected to be almond vicilin. However, this allergen was not designated by the World Health Organization/International Union of Immunological Societies. This study aimed to determine the true identity of this elusive almond allergen. cDNAs were synthesized from total RNA of the Nonpareil almond. The complete sequence of the previously reported almond allergen was determined from its coding sequence. The deduced protein was produced recombinantly and was confirmed to be a food allergen by testing with 18 almond-allergic sera. The allergen is a potential cysteine-rich antimicrobial protein with characteristic C[X]3C-[X]10-12-C[X]3C motifs of the hairpinin antimicrobial protein. This first member of a novel family of food allergens was named Pru du 8. The signature motif of the hairpinin antimicrobial protein can be found in the N-terminal region of some vicilin allergens (e.g., Ara h 1). It can also be found in the signal peptide of other vicilin allergens (e.g., Car i 2). In many species, however, vicilins do not contain such a motif, indicating that the presence of the signature motifs of the hairpinin antimicrobial protein in vicilins might be a result of translocation during evolution.

[Clinical application of pelvic floor reconstruction in extralevator abdominoperineal excision for low rectal cancer].

OBJECTIVE: To investigate the safety and clinical significance of pelvic floor reconstruction in extralevator abdominoperineal excision(ELAPE) for advanced low rectal cancer. METHODS: The clinical efficacy was retrospectively analyzed in 30 patients with low rectal cancer who underwent ELAPE from January 2013 to December 2016 in Nanjing Drum Tower Hospital, the affiliated Hospital of Nanjing University Medical School. There were 21 male patients and 9 female, with an average age of 61.7 years old. We used 13*15 cm Biodesign biologic meshes(Cook, China) for the reconstruction and the procedure involved soaking in saline solution for 5 minutes and fixation of the mesh to the cut edges of the levators by non-absorbable 2-0 sutures. A perineal drain was used and was removed when drainage was minimal. Potassium permanganate was used for hip bath after removing the stitches. The surgical procedure, postoperative complications, prognosis and follow-up of all these patients were documented. RESULTS: The operations of all patients were completed successfully. ELAPE could remove more para cancer tissues in the distant rectum. There was no rectum perforation, and the circumferential resection margins of all specimens were proved to be negative. During the follow-up of 21 months, only 2 patients suffered incision infection and healed uneventfully after strengthening the dressing. No one developed perineal breakdown, bulge or intestinal obstruction, as well as local recurrence and pelvic floor hernia. There was also no complication related to mesh. The average hospitalization time was 10 days (9-15 days). CONCLUSIONS: The ELAPE could render a low occurrence of intraoperative perforations and circumferential resection margins. Reconstruction of pelvic floor with biologic meshmight lower the complication incidences associated with the perineal region.

Extralevator abdominoperineal excision for rectal cancer with biological mesh for pelvic floor reconstruction.

GOAL: To share our experience of extra-levator abdominoperineal excision (ELAPE) for low rectal cancer, focusing on perineal repair with biological mesh. METHODS: We retrospectively analyzed medical records of all patients with low rectal cancer who underwent the ELAPE procedure using biological mesh for perineal repair at the Gastrointestinal Surgery of Nanjing Drum Power Hospital between January 2013 and September 2015. All patients were closely followed up to now. RESULTS: A total of 17 patients underwent ELAPE for low rectal cancer was screened. Of these, 15 patients had primary rectal cancer, 1 had local recurrent rectal cancer, and 1 had malignant melanoma. All patients underwent ELAPE successfully without intestinal perforation and got stage I healing in perineum wound without incision infection, dehiscence, cystocele perinealis, urethral dysfunction or intestinal obstruction. Perineum wound hematoma developed in just one patient and had successful percutaneous drainage in one week. During the follow-up, there was no recurrence, perineal hernia, sexual dysfunction, urinary retention, or bowel obstruction. Two patients described slight pain in the sacrococcygeal region without special handling. CONCLUSION: ELAPE is applicable to low rectal cancer. Biological mesh reconstruction of perineal defect seems to be safe and effective, with high patient compliance.

Inhibition effect of blunting Notch signaling on food allergy through improving TH1/TH2 balance in mice.

BACKGROUND: Notch signaling regulates proliferation, differentiation, and function of dendritic cells, T cells, and mast cells, as well as many other immune cells, which act as important parts in food allergy, Notch signaling may play an important role in food allergy. OBJECTIVE: To investigate the role of Notch signaling in IgE-mediated food allergy. METHODS: An ovalbumin-induced food allergy mouse model was built (cholera toxin as adjuvant) and Notch signaling was blunted by FLI-06 and MW167, which inhibited Notch receptor-expressing phase and the γ-secretase-affecting phase, respectively. Then food allergy indicators, including levels of serum antibodies, cytokines, and degranulation, were examined. Meanwhile, clinical features, such as vascular permeability changes, intestinal permeability changes, body temperature changes, and symptoms, were also observed. RESULTS: After blunting Notch signaling, the levels of serum ovalbumin specific IgE and IgG1 were decreased significantly, suggesting that blunting Notch signaling inhibited antibody responses. The levels of TH1 cytokines (interferon-γ) were increased significantly, whereas the levels of TH2 cytokines (interleukin-4, -5, and -13) were decreased significantly, suggesting TH2 polarization was suppressed after blunting Notch signaling. The expression of T-bet was significantly increased, whereas the expression of Gata-3 was significantly reduced in both messenger RNA and protein levels, indicating TH2 polarization was inhibited and TH1 polarization was enhanced after blunting Notch signaling. Moreover, allergic clinical features of mice were alleviated after blunting Notch signaling. CONCLUSION: Food allergy was inhibited by blunting Notch signaling through suppressing TH2 polarization, enhancing TH1 cell differentiation and promoting TH1/TH2 balance in mice. Notch signaling plays a key role in IgE-mediated food allergy.

[Analysis of clinicopathological features and prognosis in 30 cases with multifocal gastric cancer].

OBJECTIVE: To study the clinicopathological features and prognosis of multifocal gastric cancer. METHODS: Clinicopathological data of 30 cases with multifocal gastric cancer from January 2003 to March 2014 in our department were retrospectively analyzed. Random selection of 100 cases of single focal gastric cancer patients admitted in the same period was used as control group. Clinicopathological features and prognosis were compared between two groups. RESULTS: Six(20.0%) multifocal gastric cancer patients had 3 or more focuses with different differentiation degrees at the same time. The age of multifocal gastric cancer patients was younger than that of single focal gastric cancer [(56.8±16.4) year vs. (63.3±10.8) year, P<0.05]. The TNM stage of multifocal gastric cancer was mainly stage I((73.3%, 22/30), and the TNM stage of single focal gastric cancer was mainly stage III((64.0%, 64/100). As compared to single focal gastric cancer group, multifocal gastric cancer group had smaller tumors, lower ratio of nerve invasion and lymphatic vascular invasion(all P<0.01). Five-year survival rate was higher in multifocal gastric cancer group as compared to single focal gastric cancer group(76.0% vs. 48.8%, P<0.05). The prognosis of multifocal gastric cancer patients was associated with tumor size, nerve invasion, vascular invasion, depth of tumor invasion, lymphatic metastasis and TNM staging(all P<0.05), which was similar to single focal gastric cancer patients. Differences of 5-year survival rate under various clinicopathological conditions were not significant between two groups(all P>0.05). CONCLUSION: Multifocal gastric cancer patients have earlier staging tumor and overall advantage as compared to those with single focal gastric cancer.

Different immunology mechanisms of Phellinus igniarius in inhibiting growth of liver cancer and melanoma cells.

To assess inhibition mechanisms of a Phellinus igniarius (PI) extract on cancer, C57BL/6 mice were orally treated with PI extractive after or before implanting H22 (hepatocellular carcinoma ) or B16 (melanoma) cells. Mice were orally gavaged with different doses of PI for 36 days 24h after introduction of H22 or B16 cells. Mice in another group were orally treated as above daily for 42 days and implanted with H22 cells on day 7. Then the T lymphocyte, antibody, cytokine, LAK, NK cell activity in spleen, tumor cell apoptosis status and tumor inhibition in related organs, as well as the expression of iNOS and PCNA in tumor tissue were examined. The PI extract could improve animal immunity as well as inhibit cancer cell growth and metastasis with a dose-response relationship. Notably, PI's regulation with the two kinds of tumor appeared to occur in different ways, since the antibody profile and tumor metastasis demonstrated variation between animals implanted with hepatocellular carcinoma and melanoma cells.