Does pasteurization inactivate bird flu virus in milk?

Recently, an outbreak of highly pathogenic avian influenza A (H5N1), which carries the clade 2.3.4.4b hemagglutinin (HA) gene and has been prevalent among North American bird populations since the winter of 2021, was reported in dairy cows in the United States. As of 24 May 2024, the virus has affected 63 dairy herds across nine states and has resulted in two human infections. The virus causes unusual symptoms in dairy cows, including an unexpected drop in milk production, and thick colostrum-like milk. Notably, The US Food and Drug Administration reported that around 20% of tested retail milk samples contained H5N1 viruses, with a higher percentage of positive results from regions with infected cattle herds. Data are scant regarding how effectively pasteurization inactivates the H5N1 virus in milk. Therefore, in this study, we evaluated the thermal stability of the H5 clade 2.3.4.4b viruses, along with one human H3N2 virus and other influenza subtype viruses, including H1, H3, H7, H9, and H10 subtype viruses. We also assessed the effectiveness of pasteurization in inactivating these viruses. We found that the avian H3 virus exhibits the highest thermal stability, whereas the H5N1 viruses that belong to clade 2.3.4.4b display moderate thermal stability. Importantly, our data provide direct evidence that the standard pasteurization methods used by dairy companies are effective in inactivating all tested subtypes of influenza viruses in raw milk. Our findings indicate that thermally pasteurized milk products do not pose a safety risk to consumers.

Development of a Specific Aptamer-Modified Nano-System to Treat Esophageal Squamous Cell Carcinoma.

Esophageal squamous cell carcinoma (ESCC) is a prevalent gastrointestinal cancer characterized by high mortality and an unfavorable prognosis. While combination therapies involving surgery, chemotherapy, and radiation therapy are advancing, targeted therapy for ESCC remains underdeveloped. As a result, the overall five-year survival rate for ESCC is still below 20%. Herein, ESCC-specific DNA aptamers and an innovative aptamer-modified nano-system is introduced for targeted drug and gene delivery to effectively inhibit ESCC. The EA1 ssDNA aptamer, which binds robustly to ESCC cells with high specificity and affinity, is identified using cell-based systematic evolution of ligands by exponential enrichment (cell-SELEX). An EA1-modified nano-system is developed using a natural egg yolk lipid nanovector (EA1-EYLNs-PTX/siEFNA1) that concurrently loads paclitaxel (PTX) and a small interfering RNA of Ephrin A1 (EFNA1). This combination counters ESCC's proliferation, migration, invasion, and lung metastasis. Notably, EFNA1 is overexpressed in ESCC tumors with lung metastasis and has an inverse correlation with ESCC patient prognosis. The EA1-EYLNs-PTX/siEFNA1 nano-system offers effective drug delivery and tumor targeting, resulting in significantly improved therapeutic efficacy against ESCC tumors. These insights suggest that aptamer-modified nano-systems can deliver drugs and genes with superior tumor-targeting, potentially revolutionizing targeted therapy in ESCC.

Integrated bioinformatics analysis of microarray data from non-small cell lung cancer.

Non-small cell lung cancer (NSCLC), with its high mortality rate, lack of early diagnostic markers and prevention of distant metastases are the main challenges in treatment. To identify potential miRNAs and key genes in NSCLC to find new biomarkers and target gene therapies. The GSE102286, GSE56036, GSE25508, GSE53882, GSE29248 and GSE101929 datasets were obtained from the Gene Expression Omnibus (GEO) database and screened for differentially co-expressed miRNAs (DE-miRNAs) and lncRNAs (DElncRs) by GEO2R and R software package. Pathway enrichment analysis of DE-miRNAs-target genes was performed by String and Funrich database to construct protein-protein interaction (PPI) and competing endogenous RNA (ceRNA) network and visualized with Cytoscape software. Nineteen co-expressed DE-miRNAs were screened from five datasets. The 7683 predicted up- and down-regulated DE-miRNAs-target genes were significantly concentrated in cancer-related pathways. The top 10 hub nodes in the PPI were identified as hub genes, such as MYC, EGFR, HSP90AA1 and TP53, MYC, and ACTB. By constructing miRNA-hub gene networks, hsa-miR-21, hsa-miR-141, hsa-miR-200b and hsa-miR-30a, hsa-miR-30d, hsa-miR-145 may regulate most hub genes and hsa-miR-141, hsa-miR-200, hsa-miR-145 had higher levels in the miRNA and ceRNA regulatory networks, respectively. In conclusion, the identification of hsa-miR-21, hsa-miR-141, hsa-miR-200b hsa-miR-30a, hsa-miR-30d and hsa-miR-145 provides a new theoretical basis for understanding the development of NSCLC.

Genomic Sequencing as a First-Tier Screening Test and Outcomes of Newborn Screening.

Importance: Newborn screening via biochemical tests is in use worldwide. The availability of genetic sequencing has allowed rapid screening for a substantial number of monogenic disorders. However, the outcomes of this strategy have not been evaluated in a general newborn population. Objective: To evaluate the outcomes of applying gene panel sequencing as a first-tier newborn screening test. Design, Setting, and Participants: This cohort study included newborns who were prospectively recruited from 8 screening centers in China between February 21 and December 31, 2021. Neonates with positive results were followed up before July 5, 2022. Exposures: All participants were concurrently screened using dried blood spots. The screen consisted of biochemical screening tests and a targeted gene panel sequencing test for 128 conditions. The biochemical and genomic tests could both detect 43 of the conditions, whereas the other 85 conditions were screened solely by the gene panel. Main Outcomes and Measures: The primary outcomes were the number of patients detected by gene panel sequencing but undetected by the biochemical test. Results: This study prospectively recruited 29 601 newborns (15 357 [51.2%] male). The mean (SD) gestational age was 39.0 (1.5) weeks, and the mean (SD) birth weight was 3273 (457) g. The gene panel sequencing screened 813 infants (2.7%; 95% CI, 2.6%-2.9%) as positive. By the date of follow-up, 402 infants (1.4%; 95% CI, 1.2%-1.5%) had been diagnosed, indicating the positive predictive value was 50.4% (95% CI, 50.0%-53.9%). The gene panel sequencing identified 59 patients undetected by biochemical tests, including 20 patients affected by biochemically and genetically screened disorders and 39 patients affected by solely genetically screened disorders, which translates into 1 out of every 500 newborns (95% CI, 1/385-1/625) benefiting from the implementation of gene panels as a first-tier screening test. Conclusions and Relevance: In this cohort study, the use of gene panel sequencing in a general newborn population as a first-tier screening test improved the detection capability of traditional screening, providing an evidence-based suggestion that it could be considered as a crucial method for first-tier screening.

QTL Mapping and Genome-Wide Association Study Reveal Genetic Loci and Candidate Genes Related to Soluble Solids Content in Melon.

Melon (Cucumis melo L.) is an economically important Cucurbitaceae crop grown around the globe. The sweetness of melon is a significant factor in fruit quality and consumer appeal, and the soluble solids content (SSC) is a key index of melon sweetness. In this study, 146 recombinant inbred lines (RILs) derived from two oriental melon materials with different levels of sweetness containing 1427 bin markers, and 213 melon accessions containing 1,681,775 single nucleotide polymorphism (SNP) markers were used to identify genomic regions influencing SSC. Linkage mapping detected 10 quantitative trait loci (QTLs) distributed on six chromosomes, seven of which were overlapped with the reported QTLs. A total of 211 significant SNPs were identified by genome-wide association study (GWAS), 138 of which overlapped with the reported QTLs. Two new stable, co-localized regions on chromosome 3 were identified by QTL mapping and GWAS across multiple environments, which explained large phenotypic variance. Five candidate genes related to SSC were identified by QTL mapping, GWAS, and qRT-PCR, two of which were involved in hydrolysis of raffinose and sucrose located in the new stable loci. The other three candidate genes were involved in raffinose synthesis, sugar transport, and production of substrate for sugar synthesis. The genomic regions and candidate genes will be helpful for molecular breeding programs and elucidating the mechanisms of sugar accumulation.

Nanoliter atmospheric pressure photoionization-mass spectrometry for direct bioanalysis of polycyclic aromatic hydrocarbons.

Polycyclic aromatic hydrocarbons (PAHs) are a class of low-polarity environmental contaminants that have severe carcinogenic effects and have drawn worldwide attention. However, there remain challenges for current mass spectrometric ionization techniques in the analysis of low-polarity compounds in small-volume biosamples, such as single cells. In this work, we developed a nanoliter atmospheric pressure photoionization (nano-APPI) source and optimized its parameters for the detection of PAHs in small-volume samples. We evaluated the ionization performance of the source in direct and auxiliary gas-assisted photoionization modes and analyzed different PAH compounds as well as spiked biosamples. By combining the advantages of nano-electrospray ionization (nano-ESI) and atmospheric pressure photoionization (APPI), our newly developed nano-APPI source achieved high sensitivity for the analysis of PAHs down to the fmol level. Compared to conventional atmospheric pressure chemical ionization (APCI), the detection limit of PAHs was increased by 1-2 orders of magnitude. By optimizing various parameters, we achieved highly efficient ionization of PAHs, effective analysis of PAHs in mixed components, and sensitive detection of low-abundance PAHs in single-cell samples. Our optimized nano-APPI source was successfully applied for the sensitive analysis of PAHs in complex biological samples. Based on our study, we believe that nano-APPI holds great promise for toxicological studies on complex biological samples. The present work has implications for improving the detection sensitivity of low-polarity environmental contaminants and advancing the field of MS-based analysis of small-volume biosamples.

Investigation on the binary ionization choices for large conjugated amines during electrospray ionization.

RATIONALE: Generally, amines form protonated cations ([M + H]+ ) in positive polarity during electrospray ionization (ESI). However, it was found that large conjugated amines (LCAs) had binary ionization choices of generating either radical cations (M•+ ) or [M + H]+ during ESI. Investigation on the mechanism would further our understanding of ESI. METHODS: In this work, the binary ionization behavior of LCAs was reported and studied. Internal factors (functional groups and sizes of conjugated systems) and external factors (solvent type, flow rate, and electrode position) were systematically investigated and discussed. RESULTS: For the internal factors, electron-donating groups and large conjugated structures of LCAs were conducive to the generation of M•+ . For the external factors, aprotic solvent, higher flow rate, and shorter distance from the electrode to the spray cone facilitated the formation of M•+ but hampered the generation of [M + H]+ . CONCLUSION: The present study illustrated that the formations of M•+ and [M + H]+ for LCAs were two independent processes. The M•+ cations of LCAs were formed on the surface of the electrode through electrochemical oxidation, whereas the [M + H]+ cations were generated following the typical ESI evolution process. By regulating the external factors, the ionization results of LCAs could be well modulated.

Melatonin regulates trophoblast pyroptosis, invasion and migration in preeclampsia by inhibiting HtrA1 transcription through the microRNA-520c-3p/SETD7 axis.

OBJECTIVE: Melatonin has an inhibitory effect on preeclampsia (PE). This study was launched to explore the way that melatonin regulated trophoblast migration, invasion, and pyroptosis in PE and to provide new ideas for the diagnosis and treatment of PE. METHODS: Expression levels of melatonin receptors (MT1 and MT2), microRNA (miR)-520c-3p, SETD7, and HtrA1 in placental tissues and HTR8/SVneo cells were measured by RT-qPCR and Western blot. Scratch, Transwell, and Western blot assays were performed to detect migration, invasion, and pyroptosis of hypoxia/reoxygenation (H/R)-treated HTR8/SVneo cells. Dual-luciferase reporter assay was utilized to verify the targeting relationship between miR-520c-3p and SETD7. ChIP experiment was conducted to detect the enrichment of H3K4me3 and SETD7 in HtrA1 promoter. RESULTS: Low expression of MT1, MT2, and miR-520c-3p and high expression of SETD7 and HtrA1 were observed in the placental tissues of PE patients and H/R-treated HTR8/Svneo cells. A high concentration of melatonin promoted migration and invasion and inhibited pyroptosis of PE cell models. Knockdown of miR-520c-3p, overexpression of SETD7, or overexpression of HtrA1 impaired migration and invasion and accelerated pyroptosis of H/R-treated HTR8/SVneo cells, but these outcomes could be reversed by treatment with 1000 µM melatonin. miR-520c-3p targeted SETD7 which promoted histone methylation in the promoter region of HtrA1. CONCLUSION: Melatonin may inhibit HtrA1 transcription through the miR-520c-3p/SETD7 axis to promote trophoblast invasion and migration and reduce trophoblast pyroptosis in PE.

Contrast-enhanced ultrasound targeted versus conventional ultrasound guided systematic prostate biopsy for the accurate diagnosis of prostate cancer: A meta-analysis.

BACKGROUND: Conventional transrectal ultrasonography (TRUS) guided prostate biopsy is the standard method for accurate diagnosis of prostate cancer (PCa). However, the limitations of this technique in terms of missed diagnosis cannot be ignored. Based on previous studies, contrast-enhanced ultrasound (CEUS) may be able to more distinctly detect malignant lesions with increased microvessels. Therefore, to evaluate the diagnostic efficiency and clinical application prospects of CEUS-guided prostate biopsy for patients with suspected PCa, we performed a meta-analysis comparing CEUS-targeted with TRUS-guided systematic biopsy. METHODS: A systematic search of PubMed, Web of Science, Embase and CNKI was performed up to March, 2022 for the relevant published studies. After data extraction and quality assessment, meta-analysis was performed using the RevMan 5.3 software. RESULTS: The results showed that the overall sensitivity was higher for CEUS targeted biopsy than systematic biopsy (P = .03), so was the accuracy (P = .03). However, significant heterogeneity and inconsistent results from certain subgroup analyses challenged the validity of the results. Meanwhile, CEUS yielded a much higher sensitivity in patients with prostate specific antigen (PSA) level of 4 to 10 ng/mL (P = .007). On the other hand, the positive rate of each core (P < .001) and the detection rate of clinically significant PCa (P = .006) were significantly improved using CEUS. CONCLUSION: CEUS showed the advantage of a higher detection rate of clinically significant PCa, which might provide more specific indications for subsequent treatment. More feasible, real-time data are required to confirm our findings.

Ammonium Bicarbonate Significantly Accelerates the Microdroplet Reactions of Amines with Carbon Dioxide.

The reactions between amines and carbon dioxide (CO2) are among the most commonly used and important carbon fixation reactions at present. Microdroplets generated by electrospray ionization (ESI) have been proved to increase the conversion ratio (RC) of amines. In this work, we confirmed that the presence of ammonium bicarbonate (NH4HCO3) in ESI microdroplets significantly increased the RC of amines. The RC went up remarkably with the increase in the concentration of NH4HCO3 from 0.5 to 20 mM. The RC of N,N-dibutyl-1,3-propanediamine (DBPA) reached 93.7% under 20 mM NH4HCO3, which was significantly higher than previous reports. The rise in RC became insignificant when the concentration of NH4HCO3 was increased beyond 20 mM. Further investigations were made on the mechanism of the phenomenon. According to the results, it was suggested that NH4HCO3 decomposed into CO2 and formed microbubbles within the microdroplets of ESI. The microbubbles acted as direct internal CO2 sources. The conversion reactions occurred at the liquid-gas interface. The formation of CO2 microbubbles remarkably increased the total area of the interface, thus promoting the conversion reactions. 13C-labeled experiments confirmed that NH4HCO3 acted as an internal CO2 source. Factors that influenced the RC of the reaction were optimized. Pure water was proved to be the optimal solvent. Lower temperature of the mass spectrometer's entrance capillary was beneficial to the stabilization of the product carbamic acids. The sample flow rate of ESI was crucial to the RC. It determined the initial sizes of the microdroplet. Lower flow rates ensured higher RC of amines. The present work implied that NH4HCO3 could be a superior medium for CO2 capture and utilization. It might offer an alternative choice for future CO2 conversion research studies. In addition, our study also provided evidence that NH4HCO3 decomposed and generated microbubbles in the droplets during ESI. Attention should be paid to this when using NH4HCO3 as an additive in mass spectrometry-based analysis.

Strong Acid Anions Significantly Increasing the Charge State of Proteins during Electrospray Ionization.

Regulation of protein's charge state in electrospray is of great importance to the analysis of proteins. Different methods have been developed so far to increase the charge state of proteins. In this work, we investigated the influence of different anions on the charge state of proteins. Both strong acid anions and weak acid anions were taken into consideration. The results showed that the presence of 5 mM strong acid anions in acidic solutions could significantly increase the charge state of proteins. In comparison, weak acid anions with the same concentration in solution had little impact on the charge state of proteins. The species of the cations in the samples had very limited influence on the charge state. The presence of a certain amount of acid in sample solution was critical to the effect of strong acid anions. Almost no increase of the charge state was observed when no acid was added to the samples. However, remarkable increase of the charge state of myoglobin (Mb) was observed when 0.001% (v/v) acetic acid (HAc) was added to the sample together with 5 mM sodium chloride (NaCl). A higher concentration of acid in samples would further enhance the effect of strong acid anions on the increase of the charge state. Further investigations into the mechanism revealed that the effect of the strong acid anions on the charge state of proteins was based on the unfolding of the protein molecules during electrospray ionization (ESI). The interactions among H+, anions, and protein molecules were so strong that it caused the unfolding of protein molecules and resulted in the increasing of proteins' charge states. The key factor that made strong acid anions and weak acid anions different in the results was the hydrolysis of the weak acid anions in acidic solutions. The present work furthers our understanding about electrospray, as well as the regulation of protein charge state. The presence of strong acid anions in acidic solutions can significantly influence the charge state of proteins in electrospray. Attention should be paid to this when regulating the charge state of proteins.

Preparation, Characterization, and Performance Control of Nanographitic Films.

Using methane as a carbon source, low-dimensional carbon nanomaterials were obtained in this work. The films were deposited directly on glass substrates by radio frequency plasma-enhanced chemical vapor deposition (RF-PECVD). The configuration and compositions of this nanographite films were identified by X-ray photoelectron spectroscopy (XPS) as carbon in sp2 bonding form. Raman spectral characterization verified the configuration of the films to be hexatomic ring of carbon atoms. As a result, they were found to be nanographite films (NGFs). Also, the atomic force microscopy (AFM) topography and Raman spectra of different areas demonstrated the diversity of the films at the nano scale. The high light-transmitting and electron mobility indicated that the NGFs possessed excellent optic-electronic properties and could be used as good photoelectrical function materials. Furthermore, the physical and chemical growth mechanism of NGFs were analyzed by PECVD. NGFs could be obtained in a controlled process by modulating the growth conditions. In this work, the complicated transfer process commonly used for optoelectronic devices could be avoided. Also, by growing the films directly on a glass substrate, the quality degradation of the film was not a problem. This work can further promote the development of next-generation electronic or optoelectronic function materials, especially for their application in transparent conductive electrode fields.

Hepatic expression of cannabinoid receptors CB1 and CB2 correlate with fibrogenesis in patients with chronic hepatitis B.

BACKGROUND: The endocannabinoid system is involved in the pathogenesis of liver fibrosis. However, most of the findings in this area have come from experimental studies in animal models or clinical trials on chronic hepatitis C. The roles of cannabinoid receptor 1 (CB1) and cannabinoid receptor 2 (CB2) in hepatofibrosis in patients with chronic hepatitis B (CHB) have not been studied fully. This study aimed to explore the relationship between liver fibrosis and the expression of CB1 and CB2 in patients with CHB. METHODS: Eighty liver biopsy specimens from patients with CHB (52 male, 28 female) were analyzed in this study. Fibrosis was staged on a scale of 1 to 4 (F1 to F4, with F4 defining cirrhosis). There were 20 samples for each fibrosis stage. The expression of hepatic alpha-smooth muscle actin (α-SMA), CB1, and CB2 was detected by immunohistochemistry. RESULTS: Hepatic CB1 and CB2 were expressed in all patients with CHB. The degree of fibrosis was significantly associated with the increased expression of CB1 and CB2 in CHB. Furthermore a significant increase in cells positive for both CB1 and CB2 was detected in stage 3 and stage 4 disease compared to stage 1 and stage 2 disease. There was a strong positive association between CB1 expression and α-SMA expression. Moreover, double immunofluorescence staining for CB1 and α-SMA demonstrated that activated hepatic stellate cells (HSCs) express CB1. CONCLUSIONS: The hepatic expression of CB1 and CB2 plays an important role during the progression of fibrosis induced by CHB. Endogenous activation of CB1 receptors in patients with CHB enhances fibrogenesis by direct effect on activated HSCs.

Fatty Acid-Binding Protein 4 mediates apoptosis via endoplasmic reticulum stress in mesangial cells of diabetic nephropathy.

Type 2 diabetes is characterized by hyperglycemia and deregulated lipid metabolism with increased plasma non-esterified fatty acids (NEFA). Apoptosis of glomerular cells is a hallmark in diabetic glomerulosclerosis. Fatty acid-binding protein 4 (FABP4), a carrier protein for fatty acids, has been linked to diabetes and diabetic nephropathy (DN). Here we aimed to investigate the link between FABP4 and apoptosis in diabetic glomerulosclerosis. We first evaluated the presence of FABP4 and ER stress markers as well as apoptosis-related proteins in renal biopsies of patients with DN. Then we used FABP4 inhibitor BMS309403 or siRNA to further investigate the role of FABP4 in ER stress and apoptosis induced by NEFA or high glucose in cultured human mesangial cells (HMCs). We found FABP4 was expressed mainly in glomerular mesangial cells of the human renal biopsies and the glomerular FABP4 was increased in renal biopsies of DN. The up-regulation of FABP4 was accompanied with increased glucose-regulated protein 78 (GRP78) and Caspase-12 as well as down-regulated B-cell CLL/lymphoma 2 (Bcl-2) in glomeruli. Along with the induction of FABP4 and apoptosis, GRP78 and its three sensors as well as C/EBP homologous protein (CHOP) and Caspase-12 were induced in HMCs treated with NEFA or high glucose and these responses were attenuated or even abrogated by treating with FABP4 inhibitor or FABP4 siRNA. Ultrastructure observation confirmed the lipotoxicity of oleic acid by showing the morphological damage in HMCs. Our data suggest that FABP4 in glomerular mesangial cells is up-regulated in DN and FABP4 mediates apoptosis via the ER stress in HMCs.

[Ecological engineering experiment for Jinshan Lake in Zhenjiang base on techniques of immobilized nitrogen cycling bacteria].

Nitrogen cycling bacteria, including ammonifying, nitrobacteria, nitrosobacteria and denitrifying bacteria were screened, carrier was made and immobilized nitrogen cycling bacteria (INCB) was prepared. The results demonstrated that ammonifying, nitrobacteria, nitrosobacteria and denitrifying bacteria were increased markedly in the experimental areas and root zone of aquatic plants by releasing of INCB. The results also showed that the average removal efficiencies for total N (TN), NH4(+) -N, NO3(-) -N and NO2(-) -N were 44.70%, 67.17%, 31.79% and 74.21%, respectively. Furthermore, NH4(+) -N, total N (TN) reached the National Standard II and IV for surface water, respectively. With INCB, local lake water quality could improve. Therefore, the technique of INCB could play an important role for remedying and rehabilitating in desertification water.