Establishment and validation of a 5-factor diagnostic model for obstructive and non-obstructive azoospermia based on routine clinical parameters.

Azoospermia is a serious leading male-factor cause of infertility in couples of childbearing age. The two main azoospermia types, obstructive (OA) and non-obstructive (NOA) azoospermia, differ in their treatment approaches. Therefore, their clinical diagnosis is extremely important, requiring an accurate, efficient, and easy-to-use diagnostic model. This retrospective observational study included 707 patients with azoospermia treated between 2017 and 2021, 498 with OA, and 209 with NOA. Hematological and seminal plasma parameters, hormone levels, and testicular volume were used in logistic regression analysis to evaluate and compare their diagnostic performance, results showed that the optimal diagnostic model is constructed by five variables including semen volume, semen pH, seminal plasma neutral α-glucosidase activity, follicle-stimulating hormone in the serum, and testicular volume, compared with follicle-stimulating hormone-based and testicular volume-based models. The 5-factor diagnostic model had an accuracy of 90.4%, sensitivity of 96.4%, positive predictive value of 90.6%, negative predictive value of 89.8%, and area under the curve of 0.931, all higher than in the other two models. However, its specificity (76.1%) was slightly lower than in the other models. Meantime, the internal 5-fold cross-validation results indicated that the 5-factor diagnostic model had a good clinical application value. This study established an accurate, efficient, and relatively accessible 5-factor diagnostic model for OA and NOA, providing a reference for clinical decision-making when selecting an appropriate treatment.

pH-induced interaction mechanism of zearalenone with zein: Binding characteristics, conformational structure and intermolecular forces.

Zein-bound zearalenone (ZEN) complexes are naturally existed in maize by their spontaneous interaction, which significantly impacts the risk assessment of ZEN. Additionally, the pH levels in processing could affect the binding or release of zein-bound ZEN. In this study, pH-induced interaction mechanism of ZEN with zein were studied. Results showed that the acid conditions increased the binding constant (Ka) from 3.46 to 10.0 × 104 L/mol, binding energy from -17.38 to -43.49 kJ mol-1. By increasing hydrophobic interaction and hydrogen bond of ZEN with zein, the binding of ZEN with zein was promoted, forming zein-bound ZEN. Whereas, alkaline conditions decreased the Ka to 1.45 × 104 L/mol and binding energy to 148.48 kJ mol-1, weakened ZEN-zein interaction and stretched zein molecules, resulting the release of ZEN from zein. This study could provide important theoretical basis for perfecting risk assessment and controlling zein-bound ZEN during processing.

Effect of modified starches on the quality of skins of glutinous rice dumplings.

This study aimed to investigate the influence of modified starches on the quality of skins of glutinous rice dumplings (SGRDs), including changes in textural properties, pasting parameters, microstructure, color, transparency, and sensory quality. The results showed that the addition of a single acetylated-modified cassava or potato starch or composite modified cassava and potato starch in a ratio of 2:1 can improve the quality of SGRDs. The springiness and lightness of SGRDs increased, and the transparency increased from 3.22 % to 6.18 %. The cooked samples had delicate mouth-feel, uniform color and luster, good transparency, no depression, and low weight loss and did not stick to the teeth. Moreover, the total consumer acceptability score increased from 60.67 to 89.33, indicating that these products were widely accepted by consumers. However, the addition of hydroxypropyl-modified cassava starch or its composite with other two modified starches had no apparent effect on the quality of SGRDs. In conclusion, the quality of SGRDs were significantly improved by the addition of single or composite acetylated-modified starches. This study provides a theoretical basis for improving the quality of SGRDs.

Emerging Zein-Bound Zearalenone in Maize: Thermal-Induced Mechanism of Binding or Releasing.

The emerging zein-bound zearalenone (ZEN) in maize could affect its nutrition and health. Besides, thermal processing could affect the zein-ZEN interaction, causing the binding or release of ZEN. To control the harm of zein-bound ZEN on the quality of maize, the thermal-induced mechanism of binding or releasing of zein-bound ZEN were studied. Results showed that thermal processing decreased the binding constant from 1.70 to 0.27 × 104 L mol-1, and binding energy from -78.41 to -32.51 kJ mol-1, with the decreased hydrogen bonds, hydrophobic, and electrostatic interactions of ZEN with Leu81 and Arg85, Val125, Ala129, and Gln132. Furthermore, thermal processing destroyed the interactions among zein molecules and caused the unwinding of zein, releasing the ZEN from the hydrophobic cavity of zein. This paper provided theoretic insights into the heat-induced binding/releasing mechanism of ZEN with zein, which helped to perfect the exposure risk evaluation of ZEN (including free and zein-bound ZEN) in maize-based products.

Comparison of the performance of two automatic cell morphology analyzers for peripheral-blood leukocyte morphology analysis: Mindray MC-100i and Sysmex DI-60.

INTRODUCTION: To compare the morphological classification ability of peripheral-blood leukocytes of the automatic cell morphology analyzers MC-100i and DI-60. METHODS: (1) MC-100i and DI-60 were used to analyze leukocytes in 432 venous blood samples collected from three tertiary hospitals across China. The preclassification results were compared with the results reported by senior morphological experts (postclassification results) to evaluate the accuracy, sensitivity, specificity, and consistency of leukocyte preclassification for both instruments. (2) In 200 of the 432 blood samples, morphological experts conducted manual microscopic examination for various types of leukocytes. The correlation between the MC-100i and DI-60 leukocyte postclassification results and the expert microscopist results were analyzed. RESULTS: (1) MC-100i preclassified leukocytes and nucleated red blood cells (RBCs). Compared with the postclassification results, the total leukocyte preclassification accuracy of MC-100i was 97.16%, while that of DI-60 was 87.24%. The sensitivity of MC-100i to abnormal cells (including blasts, promyelocytes, neutrophilic myelocytes, neutrophilic metamyelocytes, reactive lymphocytes, abnormal promyelocytes, plasma cells, abnormal lymphocytes and nucleated RBCs) was 90.24%, which was significantly higher than the 50.72% sensitivity of DI-60. (2) Comparing the postclassification results with manual microscopy, except for reactive lymphocytes and basophils, the MC-100i and DI-60 results had good correlations with various leukocyte types and nucleated RBCs (r > 0.85), and MC-100i was better than DI-60 in the recognition of basophils. CONCLUSION: Both MC-100i and DI-60 have good detection ability for five normal types of leukocytes in peripheral blood. MC-100i has significantly better detection sensitivity for abnormal cells in peripheral blood than DI-60.

Effect of oil phases on the stability of myofibrillar protein microgel particles stabilized Pickering emulsions: The leading role of viscosity.

The Pickering emulsion may be restricted in the foods owing to the unreasonable use of oils. Herein, the effect of different oil phases on the stability of myofibrillar protein microgel particles stabilized Pickering emulsions was investigated. Results showed sunflower oil Pickering emulsions with high stability have the smallest droplet size (-26.17 µm). While peanut oil Pickering emulsions have the largest droplet size (-77.00 µm) and poor emulsion stability. The fatty acid analysis showed sunflower oil had low content of saturated (15.68 %) and super-long-chain (0) fatty acids, while peanut oil had high content of saturated (23.67 %) and super-long-chain (9.02 %) fatty acids, leading to a difference in viscosity. Low viscosity was more conducive to dispersing oil droplets and inhibiting the floating and gathering of droplets, thus enhancing the emulsion stability. Therefore, the oil with low content of super-long-chain and saturated fatty acids could be suitable for preparing MMP Pickering emulsions.

Matrix-associated mycotoxins in foods, cereals and feedstuffs: A review on occurrence, detection, transformation and future challenges.

Matrix-associated mycotoxins that bind with macromolecular components through covalent or non-covalent interactions easily occur in various cereals, cereal-based products, and cereal-based feedstuff. They are "masked" by macro-components, causing the underestimation of total exposure risk of mycotoxins. Most of the current reports focus on the free and modified mycotoxins, while the matrix-associated forms are ignored but still can exert toxic effects after ingestion. In this paper, current researches and future prospects of matrix-associated mycotoxins are reviewed. Especially, a focus is set on the transformation of matrix-associated mycotoxins with their free forms during metabolism and food processing. Enzymes, temperature and pH levels during food processing can induce the interconversion of matrix-associated mycotoxins with free mycotoxins. Furthermore, the analytical methods targeted on matrix-associated mycotoxins are discussed. Due to the lack of efficient methods releasing the mycotoxins from matrix, the standard analytical methods has not developed so far. Also, we further analyzed the challenges of matrix-associated mycotoxins about variety, occurrence, toxicity and transformation, exposure assessment, which contributes to establish preventive measures to control their hazards for consumers. Overall, this overview is significant for perfecting risk assessment, as well as developing effective prevention and control actions to matrix-associated mycotoxins.

Study of Competitive Displacement of Curcumin on α-zearalenol Binding to Human Serum Albumin Complex Using Fluorescence Spectroscopy.

α-zearalenol (α-ZOL) is a mycotoxin with a strong estrogen effect that affects the synthesis and secretion of sex hormones and is transported to target organs through human serum albumin (HSA). Additionally, it has been reported that curcumin can also bind to HSA with high affinity at the same binding site as α-ZOL. Additionally, several studies reported that reducing the bound fraction of α-ZOL contributes to speeding up the elimination rate of α-ZOL to reduce its hazard to organs. Therefore, to explore the influence of a nutrition intervention with curcumin on α-ZOL effects, the competitive displacement of α-ZOL from HSA by curcumin was investigated using spectroscopic techniques, ultrafiltration techniques and HPLC methods. Results show that curcumin and α-ZOL share the same binding site (subdomain IIA) on HSA, and curcumin binds to HSA with a binding constant of 1.12 × 105 M-1, which is higher than that of α-ZOL (3.98 × 104 M-1). Ultrafiltration studies demonstrated that curcumin could displace α-ZOL from HSA to reduce α-ZOL's binding fraction. Synchronous fluorescence spectroscopy revealed that curcumin could reduce the hydrophobicity of the microenvironment of an HSA-α-ZOL complex. This study is of great significance for applying curcumin and other highly active foodborne components to interfere with the toxicokinetics of α-ZOL and reduce its risk of its exposure.

Temperature and pH levels: Key factors effecting hidden/free zearalenone during maize processing.

The productions of food, agricultural products and feed involve a variety processing conditions, which can affect the release/binding of zearalenone (ZEN) with macro-components in maize, resulting in the interconversion of hidden ZEN with free ZEN. This interconversion can cause the uncertainties and individual differences in the exposure assessment. To fully overcome this uncertainties of the potential risks for humans and animals, further research regarding the key factors and rule of interconversion is needed. This study was designed to analyze the key factors that can induce the interconversion of hidden ZEN with free ZEN during maize processing. Results showed the conversion of hidden ZEN into free ZEN during thermal and alkaline processing, while it showed the conversion of free ZEN into hidden ZEN during acid processing. Furthermore, the temperature and pH-induced interconversion rules were also applicable to the practical maize processing (tortilla production). During producing tortilla, when the processing conditions of 4% lime, cooking time (60 min) and cooking temperature (100 °C) were set, the tortilla products with the lower hidden ZEN hazard could be obtained. This study presented new insights into the risk assessment of maize products as the conversion between hidden ZEN and free ZEN during processing.

Zein structure and its hidden zearalenone: Effect of zein extraction methods.

Zein, the plant protein, has received great attention in the last years. However, hidden zearalenone (ZEN), the zein-bound ZEN present in zein would have a great hazard to humans and animals. To date, the fates of hidden ZEN under different extraction methods of zein have not been clarified. In this study, the effect of zein extraction methods on conversion of free ZEN with hidden ZEN and its corresponding mechanism were studied. Results showed that the acid extraction condition promoted the formation of hidden ZEN (up to 92.03%), which was closely related to increased hydrophobic cavity of zein. However, alkaline extraction condition caused the conversion of hidden ZEN (from 58.82% to 13.33%) into free ZEN (from 41.18% to 86.67%), which was attributed to the great denaturation of zein. This study is of great significance for controlling hidden ZEN during zein extraction and maize processing.

Preclinical evidence of synergism between atovaquone and chemotherapy by AMPK-dependent mitochondrial dysfunction.

Chemoresistance has been associated with increased reliance on mitochondrial functions in many cancers, including lung cancer. Atovaquone is an anti-malaria drug and mitochondrial inhibitor. In this work, we attempted to explore whether atovaquone can be repurposed for lung cancer treatment to overcome chemoresistance. We showed that atovaquone inhibited proliferation, colony formation and survival in non-small cell lung cancer cell (NSCLC) cells. Of note, the effective dose of atovaquone was clinically achievable. Combination index value indicated that atovaquone and carboplatin were synergistic in inhibiting NSCLC. The potent efficacy of atovaquone and its synergism with chemotherapeutic drug were also demonstrated in NSCLC xenograft mice model. Mechanism studies showed that the synergism between atovaquone and carboplatin was due to atovaquone's ability in disrupting mitochondrial functions via specifically inhibiting complex III induced oxygen consumption. Subsequently, atovaquone activated AMP-activated protein kinase (AMPK) and inhibited mammalian target of rapamycin (mTOR) signaling. AMPK inhibition reversed the anti-NSCLC activity of atovaquone, suggesting that the action of atovaquone is also dependent on AMPK. Our work suggests that atovaquone is an attractive candidate for NSCLC treatment. Our findings emphasize that inhibition of mitochondrial function is a promising therapeutic strategy to enhance NSCLC chemosensitivity.

Effect of temperature and pH on the conversion between free and hidden zearalenone in zein.

Food processing might induce the transformation of hidden ZEN (zein-bound ZEN) in maize. The objective of this study was to assess the effect of processing factors on free ZEN and hidden ZEN. After zein was treated under different temperature and pH, ZEN was quantified in samples before and after in vitro digestion. The ratios of hidden to total ZEN in zein are decreased from 54.25% to 40.74% after thermal treatment and from 54.25% to 0 after alkaline treatment, respectively. Conversely, acid treatment increased the ratio of hidden to total ZEN from 54.25% to 100%. Thus, it can be concluded that thermal or alkaline condition induced the conversion of hidden ZEN to free ZEN while acid condition promoted the ZEN-zein interactions to form the hidden ZEN. Overall, temperature and pH values played a vital role in the conversion of hidden ZEN during food processing.

Effect of drying methods on the solubility and amphiphilicity of room temperature soluble gelatin extracted by microwave-rapid freezing-thawing coupling.

The effect of three drying methods (hot air, freeze and spray drying) on the solubility and amphiphilicity of gelatin were investigated and compared. Results showed spray drying gelatin (SDG) and hot air drying gelatin (HDG) showed the lowest and best solubility, respectively. This phenomenon was attributed to the degree of subunits degradation and hydrophobicity. The HDG showed an obvious degradation during the hot air drying and displayed the strongest hydrophilicity, while SDG showed a slight degradation and strongest hydrophobicity. The results of wettability showed that SDG had a better amphiphilicity (92.48°) in comparison with HDG (57.7°) and freeze drying gelatin (VDG, 77.53°), which can effectively reduce the interfacial tension of gelatin, thus significantly improving the stability of foam and emulsion (p < 0.05). These results suggested the drying methods can adjust the amphiphilicity of gelatin, and the SDG displayed a better amphiphilicity, showing good potential applications in foam and emulsion.

Integrated multi-spectroscopic and molecular modeling techniques to study the formation mechanism of hidden zearalenone in maize.

Hidden mycotoxins have been reported to be "protected" by macromolecular substances to escape routine determination, but release to free mycotoxins under gastrointestinal conditions. Nowadays, the hidden zearalenone (ZEN) that binding with macromolecular zein has been found in maize. However, the binding mechanism of ZEN with zein in maize has not been clarified. In this study, the formation of ZEN-zein complex was investigated applying ultrafiltration, multi-spectroscopic and molecular modeling techniques. The steady-state and transient fluorescence analysis suggested the ZEN could interact with zein to form the complex driven by hydrophobic force and hydrogen bonds, which is in accordance with the molecular modeling studies. The conformational changes of zein induced by binding with ZEN were revealed by Fourier transform infrared spectroscopy (FTIR) and circular dichroism (CD). Elucidating the binding mechanism between zein and ZEN could help the development of detecting hidden ZEN and guarantee the safety of maize products.

Improved solubility and interface properties of pigskin gelatin by microwave irradiation.

In this study, the microwave irradiation as a green approach was applied to improve the properties (mainly solubility and interface properties) of pigskin gelatin. The results showed that the solubility of pigskin gelatin was improved obviously at room temperature (25 °C) due to the destruction of polymer subunits. Furthermore, the exposure of more hydrophobic groups in microwave-irradiated gelatin increased its hydrophobicity, consequently improving the amphiphilic property and the interfacial properties of gelatin. The results of interface behavior showed that the interfacial tension of microwave-irradiated gelatin was reduced obviously with the extension of irradiation time (0-30 min), which is more beneficial to adsorption of gelatin molecules at the interface, thus resulting in a significant increase of adsorption rate (AP) from 56.13% (0 min) to 91.87% (30 min). Correspondingly, the foaming and emulsifying properties of gelatin were also improved significantly (p < 0.05). This study would promote the development of food-grade foam and emulsion based on pigskin gelatin by adjusting solubility and interface properties.

Anisomycin sensitizes non-small-cell lung cancer cells to chemotherapeutic agents and epidermal growth factor receptor inhibitor via suppressing PI3K/Akt/mTOR.

The poor outcomes in advanced non-small-cell lung cancer (NSCLC) necessitate new treatments. Recent studies emphasize anisomycin as a promising anti-cancer drug candidate. In this work, we systematically investigated the efficacy of anisomycin alone and its combination with the standard-of-care drugs in NSCLC. We showed that anisomycin inhibited growth, migration, and survival in NSCLC cells regardless of genetic mutation status, and to a greater extent than in normal lung epithelial cells. Isobologram analysis showed that the combination of anisomycin with cisplatin, paclitaxel, or gefitinib was synergistic in NSCLC but not normal lung cells. We further demonstrated that anisomycin inhibited NSCLC growth in mice. The combination of anisomycin with cisplatin was more effective than cisplatin alone and completely arrested NSCLC growth throughout the whole duration of treatment. JNK and p38 MAPK were not required for anisomycin's action. In contrast, anisomycin inhibits PI3K/Akt/mTOR pathway. Overexpression of constitutively active Akt reversed the pro-apoptotic effect of anisomycin. Our work demonstrates the selective anti-NSCLC activity of anisomycin via suppressing PI3K/Akt/mTOR. Our findings provide preclinical evidence to initialize the clinical trial of using anisomycin to sensitize NSCLC to current therapy.

Inhibition of eIF4E signaling by ribavirin selectively targets lung cancer and angiogenesis.

Although the introduction of immune- and targeted-therapy has improved the clinical response and outcomes, lung cancer remains a therapeutic challenge. Developing new therapeutics is necessary to improve the treatment of lung cancer. Here, we show that ribavirin, a clinically available anti-viral drug, is an attractive candidate for lung cancer treatment. We show that ribavirin is active against a panel of lung cancer cell lines regardless of molecular and cellular heterogeneity. Notably, the effective concentrations of ribavirin are clinically achievable, display minimal toxicity to normal cells and synergistic effect with paclitaxel. Its potent efficacy and synergism with chemotherapy on cancer cell, and minimal toxicity on normal cells are observed in lung xenograft mouse model. Ribavirin is also an angiogenesis inhibitor as it inhibits capillary network formation, growth and survival of human lung tumor-associated endothelial cell (HLT-EC). The mechanism studies demonstrate that ribavirin acts on lung cancer cells via suppressing eIF4E and mTOR signaling, leading to the subsequent inhibition of eIF4E-mediated protein translation. Our work suggests that ribavirin has advantage than many anti-cancer agents by targeting both tumor cells and angiogenesis. Our work also highlights the therapeutic potential of ribavirin for the treatment of lung cancer.

A simple mesoporous silica nanoparticle-based fluorescence aptasensor for the detection of zearalenone in grain and cereal products.

Zearalenone (ZEN) is a type of estrogenic mycotoxin commonly occurring in cereals. The aim of this study was to design a simple, rapid, inexpensive and ultrasensitive fluorescence assay for the determination of ZEN. Here, amino-modified mesoporous silica nanoparticles (MSNs-NH2) were synthesized to be the positive charge-rich reactor. A 6-carboxy-fluorescein-labeled aptamer (aptamer-FAM) was designed as the signal probe, ZEN-capture probe and negative charge reactor. In the absence of ZEN, the negatively charged aptamer-FAM combined with the positively charged MSNs-NH2 in an electrostatic manner. In the presence of ZEN, the fluorescence intensity in the supernatant increased significantly because the aptamer-FAM could bind to ZEN instead of MSNs-NH2. Under the optimal experimental conditions, this assay exhibited excellent specificity, repeatability and a wide linearity range of 0.005-150 ng/mL, with a detection limit of 0.012 ng/mL. Additionally, it showed high recovery (83.3-101.5%) for the spiked samples. There was no statistically significant difference in the ZEN concentrations detected by the proposed assay and HPLC in naturally contaminated samples. Overall, this design provides a new strategy for the rapid, inexpensive and sensitive detection of ZEN, and it could be applied to develop fluorometric assays for different targets by the selection of appropriate aptamers. Graphical abstract.

A novel fluorescence aptasensor based on mesoporous silica nanoparticles for selective and sensitive detection of aflatoxin B1.

Based on the mesoporous silica nanoparticles (MSN), a novel, simple and label-free aptamer biosensor was designed for the detection of aflatoxin B1 (AFB1). Here, the aptamers were used as molecular recognition probes and "gated molecules" while Rh6G was loaded into the interior of the particles as the signal probe. In the absence of AFB1, the "gate" was closed to prevent the leakage of the signal probe because of the immobilization of aptamers on the surface of MSN-NH2. With the presence of AFB1, the "gate" could be opened to release the signal probe for the specifical binding of aptamers to AFB1. Our results showed that the fluorescence intensity was positively correlated with the concentration of AFB1 (0.5-50 ng mL-1), with the detection limit as low as 0.13 ng mL-1. What's more, this design provides a new approach for rapid, sensitive and selective detection based on aptamers and it could be applied to numerous other analytes if appropriate aptamers are available.

Fluorescence Spectroscopic Investigation of Competitive Interactions between Quercetin and Aflatoxin B1 for Binding to Human Serum Albumin.

Aflatoxin B1 (AFB1) is a highly toxic mycotoxin found worldwide in cereals, food, and animal feeds. AFB1 binds to human serum albumin (HSA) with high affinity. In previous experiments, it has been revealed that reducing the binding rate of AFB1 with HSA could speed up the elimination rate of AFB1. Therefore, we examined the ability of quercetin to compete with AFB1 for binding HSA by fluorescence spectroscopy, synchronous spectroscopy, ultrafiltration studies, etc. It was shown that AFB1 and quercetin bind to HSA in the same Sudlow site Ӏ (subdomain IIA), and the binding constant (Ka) of the quercetin-HSA complex is significantly stronger than the complex of AFB1-HSA. Our data in this experiment showed that quercetin is able to remove the AFB1 from HSA and reduce its bound fraction. This exploratory work may be of significance for studies in the future regarding decreasing its bound fraction and then increasing its elimination rate for detoxification. This exploratory study may initiate future epidemiological research designs to obtain further in vivo evidence of the long-term (potential protective) effects of competing substances on human patients.