The repertoire of testicular extracellular vesicle cargoes and their involvement in inter-compartmental communication associated with spermatogenesis.

BACKGROUND: Spermatogenesis is regulated by a complex network of intercellular communication processes. Extracellular vesicles (EVs) are one of the important mediators in intercellular communication. Previous reports have demonstrated the involvement of EVs from the epididymis and prostate in sperm maturation and function. However, the presence of EVs in the testis and their potential involvement in spermatogenesis has not been explored. Here, we have established a testis dissociation protocol that allows the isolation and characterization of testicular EVs. RESULTS: We show that testicular EVs are specifically and efficiently taken up by somatic cells and germ cells, including the spermatozoa in the interstitial space and the seminiferous tubule compartments. We profiled the proteome of testicular EVs and probed the cell types that release them, revealing the potential contributions from the Leydig cells and testicular macrophages. Moreover, we sequenced the small RNA cargoes of testicular EVs and identified sets of small non-coding RNAs that were overlooked in the testis transcriptome. Selected miRNA candidates in testicular EVs were found in sperm RNA payload and demonstrated specific resistance towards ribonuclease A independent of the vesicle membrane. Small molecule inhibition of EV secretion perturbed spermatogenesis via inter-compartmental communication. CONCLUSIONS: Together, our study provides a valuable resource on the repertoire of cargoes carried by testicular EVs and uncovers a physiological function of testicular EVs in inter-compartmental communication associated to spermatogenesis.

One-Component Multichannel Sensor Array for Rapid Identification of Bacteria.

Bacterial infections routinely cause serious problems to public health. To mitigate the impact of bacterial infections, sensing systems are urgently required for the detection and subsequent epidemiological control of pathogenic organisms. Most conventional approaches are time-consuming and highly instrument- and professional operator-dependent. Here, we developed a novel one-component multichannel array constructed with complex systems made from three modified polyethyleneimine as well as negatively charged graphene oxide, which provided an information-rich multimode response to successfully identify 10 bacteria within minutes via electrostatic interactions and hydrophobic interactions. Furthermore, the concentration of bacteria (from OD600 = 0.025 to 1) and the ratio of mixed bacteria were successfully achieved with our smart sensing system. Our designed sensor array also exhibited huge potential in biological samples, such as in urine (OD600 = 0.125, 94% accuracy). The way to construct a sensor array with minimal sensor element with abundant signal outputs tremendously saves cost and time, providing a powerful tool for the diagnosis and assessment of bacterial infections in the clinic.

First-principles insights into hydrogen trapping in interstitial-vacancy complexes in vanadium carbide.

Hydrogen trapping is a key factor in designing advanced vanadium alloys and steels, where the influence of carbon vacancies is still elusive. Herein we have investigated the effect of carbon vacancies on the hydrogen trapping of defect-complexes in vanadium carbide using first-principles calculations. When a carbon vacancy is present, the second nearest neighboring trigonal interstitial is a stable hydrogen trapping site. A C vacancy enhances the hydrogen trapping ability by reducing the chemical and mechanical effects on H atom solution energy. Electronic structure analysis shows that C vacancies increase the charge density and the Bader atomic volume, leading to a lower H atom solution energy. The strength of the V-H bond is predominant in determining the hydrogen trapping ability in the presence of a C vacancy, in contrast to that of a C-H bond when the C vacancy is absent.

Correction: First-principles insights into hydrogen trapping in interstitial-vacancy complexes in vanadium carbide.

Correction for 'First-principles insights into hydrogen trapping in interstitial-vacancy complexes in vanadium carbide' by Shuai Tang et al., Phys. Chem. Chem. Phys., 2022, DOI: https://doi.org/10.1039/d2cp02425j.

Design of self-assembly dipeptide hydrogels and machine learning via their chemical features.

Hydrogels that are self-assembled by peptides have attracted great interest for biomedical applications. However, the link between chemical structures of peptides and their corresponding hydrogel properties is still unclear. Here, we showed a combinational approach to generate a structurally diverse hydrogel library with more than 2,000 peptides and evaluated their corresponding properties. We used a quantitative structure-property relationship to calculate their chemical features reflecting the topological and physicochemical properties, and applied machine learning to predict the self-assembly behavior. We observed that the stiffness of hydrogels is correlated with the diameter and cross-linking degree of the nanofiber. Importantly, we demonstrated that the hydrogels support cell proliferation in culture, suggesting the biocompatibility of the hydrogel. The combinatorial hydrogel library and the machine learning approach we developed linked the chemical structures with their self-assembly behavior and can accelerate the design of novel peptide structures for biomedical use.

Microfluidic Fabrication of Colloidal Nanomaterials-Encapsulated Microcapsules for Biomolecular Sensing.

Implantable sensors that detect biomarkers in vivo are critical for early disease diagnostics. Although many colloidal nanomaterials have been developed into optical sensors to detect biomolecules in vitro, their application in vivo as implantable sensors is hindered by potential migration or clearance from the implantation site. One potential solution is incorporating colloidal nanosensors in hydrogel scaffold prior to implantation. However, direct contact between the nanosensors and hydrogel matrix has the potential to disrupt sensor performance. Here, we develop a hollow-microcapsule-based sensing platform that protects colloidal nanosensors from direct contact with hydrogel matrix. Using microfluidics, colloidal nanosensors were encapsulated in polyethylene glycol microcapsules with liquid cores. The microcapsules selectively trap the nanosensors within the core while allowing free diffusion of smaller molecules such as glucose and heparin. Glucose-responsive quantum dots or gold nanorods or heparin-responsive gold nanorods were each encapsulated. Microcapsules loaded with these sensors showed responsive optical signals in the presence of target biomolecules (glucose or heparin). Furthermore, these microcapsules can be immobilized into biocompatible hydrogel as implantable devices for biomolecular sensing. This technique offers new opportunities to extend the utility of colloidal nanosensors from solution-based detection to implantable device-based detection.

Feasibility of four wastes to remove heavy metals from contaminated soils.

Soil washing is one of the permanent techniques to remove heavy metals, and washing agent is a key influence factor for this technique, but there is still lack of high-efficiency, eco-friendly, and inexpensive agents. In this study, four wastes including pineapple peel (PP), soybean straw (SS), broad bean straw (BBS) and tea residue (TR) were employed to remove cadmium (Cd), lead (Pb) and zinc (Zn) in contaminated soils. The Fourier transform infrared spectroscopy (FTIR) analysis indicated that hydroxyl, carboxyl, amine, carbonyl and amide groups were involved in the interaction with metal ions by complexation or ion exchange. We then investigated the influences of various conditions including washing solution concentration, pH, and washing time. The metal removal efficiencies with these agents increased as the concentration augmented from 5 to 80 g L-1, decreased or presented an asymmetric V-shaped curve with increasing pH from 2.5 to 7.5, and fit intraparticle diffusion or Elovich model with washing time increasing. PP has the highest removals for Cd (90.1%), Pb (18.6%), and Zn (15.2%) in soil A, and 85.8, 24.8, and 69.4% in soil B, respectively. The relatively high metal removal was mainly attributed to effective removal of the exchangeable and acid soluble fractions. Moreover, single washing not only lowered the potential ecological risk of the heavy metals, but moderated the effects on soil chemical properties. Therefore, PP was a feasible washing agent to remediate soils contaminated by heavy metals.

Reactive superhydrophobic surface and its photoinduced disulfide-ene and thiol-ene (bio)functionalization.

Reactive superhydrophobic surfaces are highly promising for biotechnological, analytical, sensor, or diagnostic applications but are difficult to realize due to their chemical inertness. In this communication, we report on a photoactive, inscribable, nonwettable, and transparent surface (PAINTS), prepared by polycondensation of trichlorovinylsilane to form thin transparent reactive porous nanofilament on a solid substrate. The PAINTS shows superhydrophobicity and can be conveniently functionalized with the photoclick thiol-ene reaction. In addition, we show for the first time that the PAINTS bearing vinyl groups can be easily modified with disulfides under UV irradiation. The effect of superhydrophobicity of PAINTS on the formation of high-resolution surface patterns has been investigated. The developed reactive superhydrophobic coating can find applications for surface biofunctionalization using abundant thiol or disulfide bearing biomolecules, such as peptides, proteins, or antibodies.

UV-Induced Tetrazole-Thiol Reaction for Polymer Conjugation and Surface Functionalization.

A UV-induced 1,3-dipolar nucleophilic addition of tetrazoles to thiols is described. Under UV irradiation the reaction proceeds rapidly at room temperature, with high yields, without a catalyst, and in both polar protic and aprotic solvents, including water. This UV-induced tetrazole-thiol reaction was successfully applied for the synthesis of small molecules, protein modification, and rapid and facile polymer-polymer conjugation. The reaction has also been demonstrated for the formation of micropatterns by site-selective surface functionalization. Superhydrophobic-hydrophilic micropatterns were successfully created by sequential modifications of a tetrazole-modified porous polymer surface with hydrophobic and hydrophilic thiols. A biotin-functionalized surface could be fabricated in aqueous solutions under long-wavelength UV irradiation.

Direct UV-induced functionalization of surface hydroxy groups by thiol-ol chemistry.

A novel UV-initiated surface modification method for the direct functionalization of surface hydroxy groups with thiol-containing molecules (termed "thiol-ol" modification) is described. This method is based on the oxidative conjugation of thiols to hydroxy groups. We demonstrate that different thiol-containing molecules, such as fluorophores, thiol-terminated poly(ethylene glycol) (PEG-SH), and a cysteine-containing peptide, can be attached onto the surface of porous poly(2-hydroxyethyl methacrylate-co-ethylene dimethacrylate). Direct functionalization of other hydroxy-group-bearing surfaces, fabrication of micropatterns, and double patterning have been also demonstrated using the thiol-ol method.

Synthetic BZLF1-targeted transcriptional activator for efficient lytic induction therapy against EBV-associated epithelial cancers.

The unique virus-cell interaction in Epstein-Barr virus (EBV)-associated malignancies implies targeting the viral latent-lytic switch is a promising therapeutic strategy. However, the lack of specific and efficient therapeutic agents to induce lytic cycle in these cancers is a major challenge facing clinical implementation. We develop a synthetic transcriptional activator that specifically activates endogenous BZLF1 and efficiently induces lytic reactivation in EBV-positive cancer cells. A lipid nanoparticle encapsulating nucleoside-modified mRNA which encodes a BZLF1-specific transcriptional activator (mTZ3-LNP) is synthesized for EBV-targeted therapy. Compared with conventional chemical inducers, mTZ3-LNP more efficiently activates EBV lytic gene expression in EBV-associated epithelial cancers. Here we show the potency and safety of treatment with mTZ3-LNP to suppress tumor growth in EBV-positive cancer models. The combination of mTZ3-LNP and ganciclovir yields highly selective cytotoxic effects of mRNA-based lytic induction therapy against EBV-positive tumor cells, indicating the potential of mRNA nanomedicine in the treatment of EBV-associated epithelial cancers.

Combinatorial synthesis and high-throughput screening of alkyl amines for nonviral gene delivery.

Efficient delivery of plasmid DNA and siRNA into cells is essential for biological and biomedical research. Although significant efforts have been made to develop efficient nonviral vectors, such as cationic lipids and polymers, most of the vectors require multistep synthesis, which complicates both fast structural optimizations and combinatorial synthesis of such vectors. Here, we present a facile, single-step method based on an alkylation of amines, allowing for the fast parallel synthesis of libraries of cationic lipid-like molecules (lipidoids). We exploited the method to synthesize 200 lipidoids, which were screened for their transfection efficiency in HEK293T cells. The screen resulted in about 2% of new lipidoids capable of efficient cell transfection similar or higher than the efficiency of Lipofectamine 2000. In addition, we observed an enhancement of cellular transfection by combining single- with double-chain lipidoids, which was attributed to the different roles of the single- and double-tailed lipids in the mixed liposomes.

CRISPR-Cas12-Based Diagnostic Applications in Infectious and Zoonotic Diseases.

Rapid detection of infectious and zoonotic diseases is very important for pathogen identification and infection control. Molecular diagnostic assays are well-known for high accuracy and sensitivity; however, conventional methods such as real-time PCR may require professional instruments and operations, preventing their wide applications in scenarios including animal quarantine. The recently developed CRISPR diagnostic (CRISPR-Dx) methods, employing the trans-cleavage activities of either Cas12 (e.g., HOLMES) or Cas13 (e.g., SHERLOCK), have shown great potential in rapid and convenient nucleic acid detection. Guided by specially designed CRISPR RNA (crRNA), Cas12 binds target DNA sequences and trans-cleaves ssDNA reporters, generating detectable signals, while Cas13 recognizes target ssRNA and trans-cleaves ssRNA reporters. To achieve high detection sensitivity, both HOLMES and SHERLOCK systems can be combined with pre-amplification procedures including both PCR and isothermal amplifications. Here, we present the employment of the HOLMESv2 method for convenient detection of the infectious and zoonotic diseases. Specifically, target nucleic acid is first amplified by LAMP or RT-LAMP, and the products are then detected by the thermophilic Cas12b. In addition, Cas12b reaction can be combined with LAMP amplification to achieve one-pot reaction systems. In this chapter, we provide a step-by-step description of the HOLMESv2-mediated rapid and sensitive detection of Japanese encephalitis virus (JEV), an RNA pathogen as an example.

A simple array integrating machine learning for identification of flavonoids in red wines.

Bioactive flavonoids, the major ingredients of red wines, have been proven to prevent atherosclerosis and cardiovascular disease due to their anti-inflammatory and anti-oxidant activity. However, flavonoids have proven challenging to identify, even when multiple approaches are combined. Hereby, a simple array was constructed to detect flavonoids by employing phenylboronic acid modified perylene diimide derivatives (PDIs). Through multiple non-specific interactions (hydrophilic, hydrophobic, charged, aromatic, hydrogen-bonded and reversible covalent interactions) with flavonoids, the fluorescence of PDIs can be modulated, and variations in intensity can be used to create fingerprints of flavonoids. This array successfully discriminated 14 flavonoids of diverse structures and concentrations with 100% accuracy, based on patterns in fluorescence intensity modulation, via optimized machine learning algorithms. As a result, this array demonstrated the parallel detection of 8 different types and origins of red wines with a high accuracy, revealing the excellent potential of the sensor array in food mixtures detection.

Printable superhydrophilic-superhydrophobic micropatterns based on supported lipid layers.

A new and simple method for creating superhydrophilic micropatterns on a superhydrophobic surface is demonstrated. The method is based on printing an "ink", an ethanol solution of a phospholipid, onto a porous superhydrophobic surface and, thus, is compatible with a variety of commonly available printing techniques.

Effect of IMB Model Combined with Spousal Support Breastfeeding Intervention on PBSES Score and Breastfeeding Rate of Primipara with Chronic Hepatitis B Virus Infection.

Objective: To analyze the effects of the info-motivation-behavior skills (IMB) model combined with spousal support breastfeeding intervention on breastfeeding Self-Efficacy Scale (PBSES) scores and breastfeeding rate of primiparas with chronic hepatitis B virus (HBV) infection. Methods: Seventy-four first-term pregnant women and 74 of their spouses were selected as the traditional control group by the convenience sampling method from July to September 2021 in obstetrics department of Shenzhen Third People's Hospital. 74 pregnant women with their first child and 74 spouses who had their first child checked during October to December 2021 were classified as the IMB model group. The traditional control group was applied with conventional intervention management mode, and the IMB model group was applied with intervention management mode based on IMB theory on the basis of the traditional control group. The self-efficacy scores of breastfeeding before and after intervention during pregnancy and during hospitalization were compared between the two groups, and the self-efficacy scores of paternal support for breastfeeding were compared. The exclusive breastfeeding rate of infants within 6 months and the maternal breastfeeding knowledge level of the two groups were compared, and the correlation between maternal breastfeeding self-efficacy score and feeding knowledge level was analyzed. Results: After pregnancy intervention, PBSES and FBSES-SF scores were significantly increased in both groups, and scores of scales in the IMB model group increased significantly than the traditional control group (all P < 0.05). The BSES-SF and FBSES-SF scores of the IMB model group increased significantly than the traditional control group at 3 days after delivery and at discharge (P < 0.05), and the scores of each scale at discharge in both groups increased significantly than those at 3 days after delivery (P < 0.05). The rate of exclusive breastfeeding in the IMB model group was 94.59% (70/74), and that in the traditional control group was 78.38% (58/74). There was a significant difference (χ2 = 8.325, P = 0.004). At discharge, the score of maternal breastfeeding knowledge increased significantly in both groups, and the score of the IMB model group increased significantly than that of the traditional control group (all P < 0.05). Pearson correlation coefficient was used to analyze the correlation between PBSES score, FBSES-SF score, and maternal feeding knowledge level, which showed positive correlation (all P < 0.05). Conclusion: The self-efficacy of prenatal breastfeeding in pregnant women with HBV is low, and the application of the IMB model combined with the intervention mode of spy-supported breastfeeding has positive effects on the improvement of maternal breastfeeding efficiency, breastfeeding health knowledge level, and postpartum breastfeeding rate, which is worthy of clinical promotion and application.

Machine Learning-Assisted Sensor Array Based on Poly(amidoamine) (PAMAM) Dendrimers for Diagnosing Alzheimer's Disease.

Alzheimer's disease (AD) is the most prevalent neurodegenerative disorder, and the early diagnosis of AD remains challenging. Here we have developed a fluorescent sensor array composed of three modified polyamidoamine dendrimers. Proteins of various properties were differentiated via this array with 100% accuracy, proving the rationality of the array's design. The mechanism of the fluorescence response was discussed. Furthermore, the robust three-element array enables parallel detection of multiple Aß40/Aß42 aggregates (0.5 µM) in diverse interferents, serum media, and cerebrospinal fluid (CSF) with high accuracy, through machine learning algorithms, demonstrating the tremendous potential of the sensor array in Alzheimer's disease diagnosis.

Associations between Abnormal Eating Styles and Irritable Bowel Syndrome: A Cross-Sectional Study among Medical School Students.

(1) Background: Few studies have investigated the association between eating styles and IBS. This study aimed to explore the association between abnormal eating styles and irritable bowel syndrome (IBS). (2) Methods: This cross-sectional study investigated students in China Medical University and Shenyang Medical College. Eating styles were evaluated by the Dutch Eating Behavior Questionnaire (DEBQ), and IBS was diagnosed according to Rome III criteria. Logistic regression was used to estimate odds ratios (ORs) and 95% confidence intervals (CIs). (3) Results: A total of 335 students were diagnosed with IBS. Students with the total scores in tertile 2 and 3 had 1.29 times and 2.75 times higher risk of IBS than students with the total scores in tertile 1, respectively. Simultaneously, the risk of IBS in the tertile 3 of external eating, emotional eating, and restraint eating trends was 3.87 times, 2.71 times, and 3.82 times higher than that of tertile 1, respectively. (4) Conclusions: this study showed that a high score in both total eating styles and each eating style was associated with the odds of having IBS and suggested that the psychological factors behind eating styles may play a critical role in controlling the IBS.

Recent Improvements in CRISPR-Based Amplification-Free Pathogen Detection.

Molecular diagnostic (MDx) methods directly detect target nucleic acid sequences and are therefore an important approach for precise diagnosis of pathogen infection. In comparison with traditional MDx techniques such as PCR, the recently developed CRISPR-based diagnostic technologies, which employ the single-stranded nucleic acid trans-cleavage activities of either Cas12 or Cas13, show merits in both sensitivity and specificity and therefore have great potential in both pathogen detection and beyond. With more and more efforts in improving both the CRISPR trans-cleavage efficiencies and the signal detection sensitivities, CRISPR-based direct detection of target nucleic acids without preamplification can be a possibility. Here in this mini-review, we summarize recent research progresses of amplification-free CRISPR-Dx systems and explore the potential changes they will lead to pathogen diagnosis. In addition, discussion of the challenges for both detection sensitivity and cost of the amplification-free systems will also be covered.

TB-QUICK: CRISPR-Cas12b-assisted rapid and sensitive detection of Mycobacterium tuberculosis.

OBJECTIVES: Tuberculosis (TB) remains one of the public health problems worldwide. Rapid, sensitive and cost-effective diagnosis of Mycobacterium tuberculosis (M.tb) is critical for TB control. METHODS: We developed a novel M.tb DNA detection platform (nominated as TB-QUICK) which combined loop-mediated isothermal amplification (LAMP) and CRISPR-Cas12b detection. TB-QUICK was performed on pulmonary or plasma samples collected from 138 pulmonary TB (PTB) patients, 21 non-TB patients and 61 close contacts to TB patients. Acid-fast bacillus (AFB) smear, M.tb culture and GeneXpert MTB/RIF (Xpert) assays were routinely conducted in parallel. RESULTS: By targeting M.tb IS6110, TB-QUICK platform could detect as low as 1.3 copy/µL M.tb DNA within 2 h. In pulmonary TB samples, TB-QUICK exhibited improved overall sensitivity of 86.8% over M.tb culture (66.7%) and Xpert (70.4%), with the specificity of 95.2%. More significantly, TB-QUICK exhibited a superior sensitivity in AFB-negative samples (80.5%) compared to Xpert (57.1%) and M.tb culture (46.2%). In the detection of plasma M.tb DNA by TB-QUICK, 41.2% sensitivity for AFB-positive and 31.7% for AFB-negative patients were achieved. CONCLUSION: In conclusion, TB-QUICK exhibits rapidity and sensitivity for M.tb DNA detection with the superiority in smear-negative paucibacillary TB patients. The clinical application of TB-QUICK in TB diagnosis needs to be further validated in larger cohort.