Lung cancer (LC) is one of the most common cancers and has the highest mortality rate. The risk of developing LC increases more than 7-fold in patients with chronic respiratory diseases, such as pulmonary fibrosis, leading to a worse prognosis and making treatment more complicated. Our hypothesis is that the fibrotic tumor microenvironment and cell-cell communication modulate tumor cell growth and modify their transcriptome We recreated tumor growth <italic>in vitro</italic> by culturing spheroids with non-small cell lung cancer cell lines (H23-adenocarcinoma, EBC1-squamous) alone or in multicellular spheroids co-cultured with human lung fibroblasts and macrophages under fibrotic (TGF-β, PDGF, FGF2, IL-4 and IL-13) and non-fibrotic conditions. We measured cell proliferation by flow cytometry, gene expression by bulk-RNA-sequencing and qPCR, and measured cell-cell interactions using microscopy. We observed that tumoral cells from mono- and multicellular spheroids under fibrotic conditions showed decreased proliferation in both LC lines. Fibroblasts in EBC1 co-culture and macrophages in H23 coculture reversed these effects. Transcriptomic data analysis revealed alterations due to fibrotic conditions in genes related to cell cycle and immune response such as CD73 that was significantly increased in both LC lines or PD-L1 that was upregulated in monocellular spheroids but decreased in the multicellular. Our data highlight the impact of the fibrotic microenvironment on tumoral cell functionality and its response to potential immunotherapies. For this reason, it is necessary to further study the altered pathways in tumor cells to identify new therapeutic options for this type of patients.

&lt;div&gt;Abstract&lt;p&gt;&lt;b&gt;Background:&lt;/b&gt; Previously, we showed that gene suppression commonly occurs across chromosome 2q14.2 in colorectal cancer, through a process of long-range epigenetic silencing (LRES), involving a combination of DNA methylation and repressive histone modifications. We now investigate whether LRES also occurs in prostate cancer across this 4-Mb region and whether differential DNA methylation of 2q14.2 genes could provide a regional panel of prostate cancer biomarkers.&lt;/p&gt;&lt;p&gt;&lt;b&gt;Methods:&lt;/b&gt; We used highly sensitive DNA methylation headloop PCR assays that can detect 10 to 25 pg of methylated DNA with a specificity of at least 1:1,000, and chromatin immunoprecipitation assays to investigate regional epigenetic remodeling across 2q14.2 in prostate cancer, in a cohort of 195 primary prostate tumors and 90 matched normal controls.&lt;/p&gt;&lt;p&gt;&lt;b&gt;Results:&lt;/b&gt; Prostate cancer cells exhibit concordant deacetylation and methylation of histone H3 Lysine 9 (H3K9Ac and H3K9me2, respectively), and localized DNA hypermethylation of &lt;i&gt;EN1&lt;/i&gt;, &lt;i&gt;SCTR&lt;/i&gt;, and &lt;i&gt;INHBB&lt;/i&gt; and corresponding loss of H3K27me3. &lt;i&gt;EN1&lt;/i&gt; and &lt;i&gt;SCTR&lt;/i&gt; were frequently methylated (65% and 53%, respectively), whereas &lt;i&gt;INHBB&lt;/i&gt; was less frequently methylated.&lt;/p&gt;&lt;p&gt;&lt;b&gt;Conclusions:&lt;/b&gt; Consistent with LRES in colorectal cancer, we found regional epigenetic remodeling across 2q14.2 in prostate cancer. Concordant methylation of &lt;i&gt;EN1&lt;/i&gt; and &lt;i&gt;SCTR&lt;/i&gt; was able to differentiate cancer from normal (&lt;i&gt;P&lt;/i&gt; &lt; 0.0001) and improved the diagnostic specificity of &lt;i&gt;GSTP1&lt;/i&gt; methylation for prostate cancer detection by 26%.&lt;/p&gt;&lt;p&gt;&lt;b&gt;Impact:&lt;/b&gt; For the first time we show that DNA methylation of &lt;i&gt;EN1&lt;/i&gt; and &lt;i&gt;SCTR&lt;/i&gt; promoters provide potential novel biomarkers for prostate cancer detection and in combination with &lt;i&gt;GSTP1&lt;/i&gt; methylation can add increased specificity and sensitivity to improve diagnostic potential. &lt;i&gt;Cancer Epidemiol Biomarkers Prev; 20(1); 148–59. ©2011 AACR&lt;/i&gt;.&lt;/p&gt;&lt;/div&gt;

&lt;div&gt;Abstract&lt;p&gt;Gene amplification is one of the most frequent manifestations of genomic instability in human tumors and plays an important role in tumor progression and acquisition of drug resistance. To better understand the factors involved in acquired resistance to cytotoxic drugs via gene amplification, we have analyzed the structure and dynamics of dihydrofolate reductase (&lt;i&gt;DHFR&lt;/i&gt;) gene amplification in HT29 cells treated with methotrexate (MTX). Analysis of the &lt;i&gt;DHFR&lt;/i&gt; gene amplification process shows that the amplicon exhibits a complex structure that is consistently reproduced in independent treatments. The cytogenetic manifestation of the amplification in advanced stages of the treatment may be in the form of double minutes or as a homogeneously stained region. To get insights into the mechanisms of resistance, we have also investigated the sensitization to MTX of MTX-resistant cells after drug withdrawal and reexposure to MTX. Passive loss of the DHFR amplicon by withdrawal of the drug results in MTX-sensitive cells exhibiting a substantial reduction of their capacity or even an incapacity to generate resistance when submitted to a second cycle of MTX treatment. On a second round of drug administration, the resistant cells generate a different amplicon structure, suggesting that the formation of the amplicon as in the first cycle of treatment is not feasible. These results indicate that &lt;i&gt;DHFR&lt;/i&gt; gene amplification is a “wear and tear” process in HT29 cells and that MTX-resistant cells may become responsive to a second round of treatment if left untreated during a sufficient period of time. [Mol Cancer Ther 2009;8(2):424–32]&lt;/p&gt;&lt;/div&gt;

Wilson disease is a rare inherited autosomal recessive disorder. As a consequence of genetic alterations in the ATP7B gene, copper begins to accumulate in the body, particularly in the liver and brain. Affected persons are prone to develop liver cancer and severe psychiatric and neurological symptoms. Clinically, the development of corneal Kayser-Fleischer rings and low ceruloplasmin concentrations (&lt;20 mg/dL) are indicative of Wilson disease. However, the detection of elevated hepatic copper content (&gt;250 µg/g dry weight) alone is still considered as the best but not exclusive diagnostic test for Wilson disease. Presently, specific copper stains (e.g., rhodanine) or indirect staining for copper-associated proteins (e.g., orcein) are widely used to histochemically visualize hepatic copper deposits. However, these procedures only detect lysosomal copper, while cytosolic copper is not detectable. Similarly, elemental analysis in scanning electron microscope with energy dispersive X-ray analysis (EDX) often leads to false negative results and inconsistencies. Here, we tested the diagnostic potential of laser ablation inductively-coupled mass spectrometry (LA-ICP-MS) that allows quantitative analysis of multiple elements. Comparative studies were performed in wild type and the Atp7b null mouse model. We propose LA-ICP-MS as a versatile and powerful method for the accurate determination of hepatic copper in people with Wilson disease with high spatial resolution.

OBJECTIVE: Anti-Mullerian hormone (AMH) together with luteinizing hormone (LH) and follicle-stimulating hormone (FSH) plays crucial roles in gonadal functions. However, the possible effects of GnRH on AMH via the hypothalamic-pituitary-gonadal (HPG) axis remain unexplored. We aimed to explore the changes in AMH levels after bolus GnRH stimulation and understand the relationship of AMH with FSH and LH in healthy subjects. METHODS: Thirty-one prepubertal children (15 males/16 females) and 78 adults (36 males/42 females) were included. We collected basal (0 minute) samples for determining levels of hormones. After GnRH treatment at a dose of 2.5 μg/kg body weight (maximum of 100 μg/kg body weight) intravenously, blood was collected at 30 minutes intervals for 120 minutes. Serum LH, FSH and AMH were measured by electrochemiluminometric assays. RESULTS: After injection of GnRH, AMH levels were significantly decreased in 30 minutes (P < 0.001) in all groups with parallel increase of FSH and LH. In the second 30 minutes, all hormones levels reversed. There was also a moderate correlation between AMH and FSH (r = -0.430, P < 0.001). CONCLUSIONS: GnRH lowers serum AMH levels, which have a negative correlation with the increase in gonadotrophins. These data pinpoint GnRH as an important factor of the AMH regulation, leading new opportunities for the understanding of AMH role in reproductive function and dysfunction.

Isolation barriers were examined by making all possible crosses including reciprocals among arrowleaf clover ( Trifolium vesiculosum Savi), T. mutabile Portenschl. and T. leiocalycinum Boiss. &amp; Spruner. No evidence of sterility was indicated by cross seed set. All F 1 cross seed germinated readily but yellow seedlings were produced by crosses of T. mutabile and T. leiocalycinum with T. vesiculosum . F 1 plants from reciprocal crosses of these species were normal green. The yellow condition disappeared as plants matured. Most F 1 plants from crosses between accessions produced less stainable pollen than plants from crosses within accessions. All species had 16 somatic chromosomes and chromosome associations at metaphase I of meiosis were similar for all parents and F 1 progenies. We concluded that T. mutabile and T. leiocalycinum are quite closely related and are slightly differentiated from T. vesiculosum . Therefore, the three taxa must be isolated under field conditions to prevent crossing.

Activation of hepatic stellate cells (HSCs) and their trans-differentiation towards collagen-secreting myofibroblasts (MFB) promote liver fibrosis progression. During chronic liver disease, resting HSCs become activated by inflammatory and injury signals. However, HSCs/MFB not only produce collagen, but also secrete cytokines, participate in metabolism, and have biomechanical properties. We herein aimed to characterize the heterogeneity of these liver mesenchymal cells by single cell RNA sequencing. In vivo resting HSCs or activated MFB were isolated from C57BL6/J mice challenged by carbon tetrachloride (CCl4) intraperitoneally for 3 weeks to induce liver fibrosis and compared to in vitro cultivated MFB. While resting HSCs formed a homogenous population characterized by high platelet derived growth factor receptor β (PDGFRβ) expression, in vivo and in vitro activated MFB split into heterogeneous populations, characterized by α-smooth muscle actin (α-SMA), collagens, or immunological markers. S100 calcium binding protein A6 (S100A6) was a universal marker of activated MFB on both the gene and protein expression level. Compared to the heterogeneity of in vivo MFB, MFB in vitro sequentially and only transiently expressed marker genes, such as chemokines, during culture activation. Taken together, our data demonstrate the heterogeneity of HSCs and MFB, indicating the existence of functionally relevant subsets in hepatic fibrosis.

Crosses were made between 2X and 4X red clover ( Trifolium pratense L.) plants under field cages by honey bees ( Apis mellifera L.) to investigate the feasibility of producing autotetraploids by the mechanism of unreduced gametes. Clones from three cultivars and one experimental strain were used as female parents (diploids) and plants from a nitrous oxide‐derived tetraploid population of the cultivar ‘Kenstar’ were used as the male parents. Diploid parents yielding low amounts of seed produced a high percentage of tetraploids, but those yielding large amounts of seed produced very few tetraploids. High seed set apparently via pseudo‐self‐compatibility or 2X‐2X crosses from pollen contamination, interfered with the production of seeds via unreduced gametes in some cages. The frequency of tetraploids derived from diploid female parents in other cages where monoploid pollen was apparently excluded ranged from one per 1500 to 11000 flowers. About half of the apparent tetraploids were aneuploids that could not be distinguished from euploids except by chromosome examination. The frequency of univalents, bivalents, and multivalents at metaphase I of the unreduced gamete population was similar to that of the nitrous oxide‐derived population. In comparison to the colchicine‐derived material, there were more univalents and bivalents and fewer multivalents at metaphase I of the unreduced gamete population. Further research on the exact mechanism of nitrous oxide doubling and a comparison of the three methods is necessary.

The present study investigates the dual impact of microbial biofertilizers on the phenotypic performance and rhizosphere microbiome composition in an onion crop. A pot experiment was conducted with seven treatments of microbial inoculants, such as Azotobacter, Azospirillum, Piriformospora indica , phosphate solubilizing bacteria (PSB), and control treatments with and without chemical fertilizers. The growth, physiological, and biochemical traits of onion were assessed alongside rhizospheric soil microbiome profiling using 16S rRNA metagenomic sequencing. Significant enhancement in plant height, leaf number, leaf area, chlorophyll content, photosynthetic rate, and antioxidant enzyme activity with low leaf temperature was observed in plants inoculated with Azotobacter and Azospirillum. Notably, the Azotobacter treatment yielded a significant enhancement in the bulb phenol content. Rhizosphere metagenomic analysis revealed 17 dominant phyla, with Actinobacteria (25.3%), Proteobacteria (22.2%), Firmicutes (12.8%), and Chloroflexi (11.02%) comprising over 70% of the total microbiome. Alpha and beta diversity metrics indicated that microbial inoculation, especially with Azospirillum and PSB, enriched the soil microbial community structure. Distinct clustering and correlations with specific microbial taxa such as Candidatus Nitrososphaera and Pseudomonas were observed in response to individual biofertilizer treatments. This study highlights the potential of biofertilizers not only in enhancing onion growth and development but also in modulating beneficial rhizosphere microbial communities. Integrating biofertilizers into onion production systems could reduce the dependency on chemical fertilizers and promote sustainable crop management.

OPINION article Front. Oncol., 11 August 2020Sec. Molecular and Cellular Oncology Volume 10 - 2020 | https://doi.org/10.3389/fonc.2020.01329

The onion cultivation sector in India is grappling with severe challenges, including acute water scarcity, labor shortages, and a sluggish adoption of sustainable agronomic practices. These issues collectively exacerbate the sector’s vulnerability to climate change and production uncertainties. This study critically evaluated the efficacy of an advanced irrigation management practice raised bed with drip irrigation (RBWD) and examined the socio-economic determinants influencing its adoption. Conducted in Maharashtra’s Ghod River basin during the 2022–2023 cropping seasons, the research encompassed 480 farmers equally divided between RBWD adopters and traditional flat bed with flood irrigation (FBWF) practitioners. The findings revealed that RBWD adoption led to significant water savings, averaging 24.52% during the late monsoon and 28.79% in the post-monsoon season while doubling water productivity compared to FBWF systems. Moreover, the implementation of RBWD resulted in a substantial increase in onion bulb yield, ranging from 13.70% to 23.61%, and enhanced bulb quality, particularly in the proportion of marketable bulbs. Family income, land holding, extension contacts and the information source use were the key determinants behind the adoption of the RBWD method. However, significant obstacles such as the high initial investment required for drip irrigation systems and the technical expertise needed for their operation continue to hinder wider adoption. These findings underscore a pressing need to address these barriers through policy interventions and targeted extension services.

OPINION article Front. Microbiol., 26 August 2020Sec. Virology Volume 11 - 2020 | https://doi.org/10.3389/fmicb.2020.01905