Transcript Levels Research Articles

Proto-oncogene HRAS transcript level and overall survival in stage II and III colorectal cancer.

231 Background: Mutational landscape is prognostic in colorectal cancer (CRC). Driver mutations in the RAS proto-oncogenes such as KRAS and NRAS portend poor survival outcomes, whereas HRAS mutations are extremely rare and their prognostic value remains uncertain. The prognostic implication of RAS gene activity at the transcript level is also obscure. Here, we retrospectively review stage II and III CRC tumor RNA-Seq data and demonstrate that high transcript levels of HRAS are associated with superior overall survival (OS). Methods: Tumor RNA-Seq of 734 stage II and III CRC cases and their associated clinical data were retrieved from the ORIEN AVATAR database (Aster Insights). Transcript level was determined by batch-corrected TPM (transcripts per million) of each gene, and cohort median was used as a reference to determine “high” versus “low” in subgroup analyses. The p-value and hazard ratio (HR) derived from Kaplan-Meier survival analysis are log-rank. Results: The 4-year HR for death in patients with “very high” HRAS transcript level (top 5%) was 0.15 (95% CI 0.07-0.34, p=0.03) compared to the rest, and this superior OS was independent from TNM stage or peri-operative 5-FU treatment history. In subgroup analyses, “high” HRAS expression was associated with superior OS only in patients with concomitant “low” KRAS expression in the absence of pathologic KRAS mutations in codons 12, 13 and 61 – the 5-year HR in “low” KRAS group was 0.42 (95% CI 0.18-0.96, p=0.03) compared to “high” KRAS group. NRAS transcript level did not influence OS. When further sub-grouped based on primary disease location, this superior OS was present only in the right-sided primary disease group with 5-year HR of “low” KRAS group at 0.17 (95% CI 0.03-0.99, p=0.06) compared “high” KRAS group. Conclusions: Contrary to the notion that RAS family genes are proto-oncogenic, we demonstrate that high HRAS transcript levels are associated with superior OS in stage II and III CRC. The high HRAS-associated OS benefit was most pronounced in patients with right-sided primary expressing low KRAS transcript levels in the absence of pathologic KRAS mutations. We speculate that when expressed at high levels, HRAS may counteract proto-oncogene KRAS but not oncogene KRAS at the transcript level. The potential of HRAS as a prognostic biomarker should be explored further.

Characterization of SARS-CoV-2 Entry Genes in Skeletal Muscle and Impacts of InVitro Versus InVivo Infection.

COVID-19 has been associated with both respiratory (diaphragm) and non-respiratory (limb) muscle atrophy. It is unclear if SARS-CoV-2 infection of skeletal muscle plays a role in these changes. This study sought to: 1) determine if cells comprising skeletal muscle tissue, particularly myofibres, express the molecular components required for SARS-CoV-2 infection; 2) assess the capacity for direct SARS-CoV-2 infection and its impact on atrophy pathway genes in myogenic cells; and 3) in an animal model of COVID-19, examine the relationship between viral infection of skeletal muscle and myofibre atrophy within the diaphragm and limb muscles. We used in silico bioinformatics analysis of published human single cell RNA-seq datasets, as well as direct qPCR examination of human myotubes and diaphragm biopsies, to assess expression of key genes involved in SARS-CoV-2 cellular entry. InVitro, we determined the ability of SARS-CoV-2 to directly infect myogenic cells and employed qPCR to assess the impact on muscle atrophy pathway genes (ubiquitin-proteasome, autophagy). Invivo, the diaphragm and quadriceps of Roborovski hamsters with SARS-CoV-2 respiratory infection were examined at day 3 post-inoculation to evaluate the relationship between atrophy pathway and SARS-CoV-2 transcripts by qPCR, as well as histological measurements of myofibre morphology. Angiotensin converting enzyme 2 (ACE2), the primary receptor for SARS-CoV-2, as well as cooperating proteases (furin, cathepsins B and L), are expressed by myofibres. ACE2 expression was increased 5-fold (p = 0.01) in the diaphragms of mechanically ventilated human subjects compared to controls. InVitro, a time-dependent increase of SARS-CoV-2 transcript levels was observed in myotubes directly exposed to the virus (p = 0.002). This was associated with downregulation of the ubiquitin ligase MuRF1 (by 64%, p = 0.002) and the autophagy gene LC3B (by 31%, p = 0.009). In contrast, invivo infection led to upregulation of MuRF1 in quadriceps (23-fold, p = 0.0007) and autophagy genes in both quadriceps (5.2-fold for Gabarapl1, p = 0.03; 7-fold for p62, p = 0.0002) and diaphragm (2.2-fold for Gabarapl1, p = 0.03; 2.3-fold for p62, p = 0.057). In infected hamsters the diaphragm lacked viral transcripts but exhibited atrophy (48% decrease in myofibre area; p = 0.02), whereas the quadriceps lacked myofibre atrophy despite elevated viral transcripts in the muscle. Although myogenic cells express the genes required for SARS-CoV-2 entry and can be directly infected, there was no evident relationship between viral transcript levels and manifestations of atrophy, either invitro or invivo. Our results do not support direct myofibre infection by SARS-CoV-2 as a likely cause of atrophy in COVID-19.

Xanthine oxidase-lactoperoxidase system: Dose-dependent antibacterial effects and global gene expression changes in infant oral microbiota.

Xanthine oxidase (XO) and lactoperoxidase (LPO) are highly abundant enzymes in milk. Their substrates, xanthine and thiocyanate, are found in elevated amounts in infant saliva, leading to a proposed interaction between milk and saliva referred to as the XO-LPO system. This system is suggested to generate reactive oxygen and nitrogen species with potential antibacterial effects. The antibacterial activity of the XO-LPO system was assessed on bacteria cultured from the oral cavities of five infants, where a reduction in bacterial growth rate was observed at 40µg mL-1 of each enzyme and with complete inhibition achieved at 200µg mL-1. Gene expression analysis showed that XO-LPO treatment led to downregulation of several reactive oxygen species-related genes, suggesting a transient bacterial stress response. The study also observed downregulation of key glycolytic enzymes, indicating that XO-LPO treatment affects bacterial metabolism at transcriptional level, suggesting a possible mechanism of action for the XO-LPO system. Collectively, these findings offer new insights into the XO-LPO system, revealing novel aspects of the interaction between lactation and microbiome influence.

The inhibitory effects of nimotuzumab on CD276 expression and immune escape in head and neck squamous cell carcinoma: Insights into anticancer mechanisms.

CD276 has been identified as a novel immune checkpoint, and its overexpression is associated with immune evasion and poor prognosis in various tumors, including head and neck squamous cell carcinoma (HNSCC). Nimotuzumab, a humanized anti-epidermal growth factor receptor (EGFR) monoclonal antibody, has been approved for various solid tumors. However, it remains unclear whether its anticancer efficacy involves a reduction in CD276 expression. The purpose of this study was to investigate the regulatory effects and potential mechanisms of nimotuzumab on CD276 expression both in vitro and in vivo. In a coculture system, nimotuzumab showed inhibitory effects on TGF-β-induced upregulation of CD276 at both the transcriptional and protein levels in HNSCC cell lines. Mechanistic studies revealed that nimotuzumab primarily suppressed TGF-β-induced CD276 upregulation by blocking EGFR/MEK/ERK, which was further validated by MEK and ERK inhibitors. In xenograft and mice HNSCC models, nimotuzumab exerted antitumor effects accompanied by significantly reduced CD276 expression during tumor progression. Analysis of tumor-infiltrating lymphocytes (TILs) profiles indicated that nimotuzumab orchestrated the tumor immune microenvironment (TIME) by notably increasing the frequency of T lymphocytes, including cytotoxic T lymphocytes and helper T lymphocytes, as well as macrophage cells. However, no significant changes were observed in the populations of NK cells, DC cells, and neutrophils. These findings offer new insights into the anticancer mechanisms of nimotuzumab and its underlying synergy in combined treatments with immunotherapy for HNSCC.

LncRNA MALAT1 as a potential diagnostic and therapeutic target in kidney diseases.

Long non-coding RNA (lncRNA) metastasis-associated lung adenocarcinoma transcript1 (MALAT1) has emerged as a crucial biomarker and therapeutic target for kidney diseases, including acute kidney injury (AKI), chronic kidney disease (CKD), diabetic kidney disease (DKD), lupus nephritis (LN), and renal cell carcinoma (RCC). LncRNAs are non-coding RNAs that have more than 200 nucleotides that play a crucial role in gene regulation at the post-translational stage, transcriptional, and epigenetic levels. LncRNA MALAT1 regulates gene expression and modulates cellular functions such as proliferation, inflammation, apoptosis, and fibrosis, which are key pathophysiology of kidney diseases. Overexpression of lncRNA MALAT1 has been consistently observed in kidney tissue, correlating with the severity and progression of kidney disease. In AKI, lncRNA MALAT1 exacerbates inflammation and tissue damage, contributing to disease progression. In CKD and DKD, lncRNA MALAT1 is implicated in the regulation of fibrosis by modulating key pathways, including focal adhesion kinase (FAK), toll-like receptor 4 (TLR4), NOD-like receptor protein3 (NLRP3), and nuclear factor kappa B (NF-κB), play pivotal roles in promoting disease progression. In LN, lncRNA MALAT1 has been linked to immune regulation and kidney damage, while in RCC, its role in promoting tumor growth and metastasis has been well documented. Preclinical research has demonstrated that therapeutic strategies targeting lncRNA MALAT1, such as knockdown and knockout, can reduce inflammation and fibrosis while improving kidney function. The fundamental role of lncRNA MALAT1 in kidney disease progression is yet to be fully understood. However, lncRNA MALAT1 has shown promise as a biomarker and therapeutic target to mitigate kidney disease development. This review highlights the potential of lncRNAs MALAT1 as diagnostic biomarkers and therapeutic targets, offering insights into a comprehensive approach to managing kidney diseases in the future.

STAT3-related lncRNAs in colorectal cancer progression; Special focus on immune cell's evasion.

Colorectal cancer (CRC) is globally ranked as the third leading cause of cancer-related deaths in both men and women. There is an urgent need for novel biomarkers to facilitate early diagnosis and enhance patient care, thereby improving treatment response and reducing mortality rates. Signal transducer and activator of transcription 3 (STAT3) is essential for controlling the anti-tumor immune response since it is a hub for several oncogenic signaling pathways. In the tumor environment, STAT3 is widely overactivated in both malignant and non-cancerous cells. It is involved in suppressing the expression of critical immune activation regulators and encouraging the synthesis of immunosuppressive substances. Long noncoding RNAs (lncRNAs), a kind of non-coding RNA, are critical for CRC development, apoptosis, and metastasis because they influence important signaling pathways such as STAT3 signaling and contribute to gene regulation at the epigenetic, transcriptional, and post-transcriptional levels. Moreover, lncRNAs have a significant role in modifying the TME and control the expression of important immunological checkpoints, such as PD-L1. Therefore, a comprehensive understanding of the regulatory roles of lncRNAs is crucial for identifying diagnostic, prognostic, and predictive biomarkers for CRC. Thus, the objective of the present review study is to provide a comprehensive overview of the interaction between the STAT3 signaling pathway and various lncRNAs, as well as their implications for apoptosis, metastasis, and immune evasion in CRC.

Ferritin Hinders Ferroptosis in Non-Tumorous Diseases: Regulatory Mechanisms and Potential Consequences.

Ferritin, as an iron storage protein, has the potential to inhibit ferroptosis by reducing excess intracellular free iron concentrations and lipid reactive oxygen species (ROS). An insufficient amount of ferritin is one of the conditions that can lead to ferroptosis through the Fenton reaction mediated by ferrous iron. Consequently, upregulation of ferritin at the transcriptional or posttranscriptional level may inhibit ferroptosis. In this review, we have discussed the essential role of ferritin in ferroptosis and the regulatory mechanism of ferroptosis in ferritin-deficient individuals. The description of the regulatory factors governing ferritin and its properties in regulating ferroptosis as underlying mechanisms for the pathologies of diseases will allow potential therapeutic approaches to be developed.

Effects of chronic cold stress and thermal stress on growth performance, hepatic apoptosis, oxidative stress, immune response and gut microbiota of juvenile hybrid sturgeon (Acipenser baerii ♀ × A. schrenkii ♂)

The current study was conducted to investigate the effects of chronic cold stress and thermal stress on the growth performance, hepatic oxidative status, immune response, apoptosis and gut microbiota in juvenile hybrid sturgeon. The fish (initial mean weight: 21.4 ± 0.3 g) was reared at three temperatures (14 °C, 22 °C, and 30 °C) for 16 d, which were termed as low temperature group (LT), moderate temperature group (MT), and high temperature group (HT), respectively, and the second group was regarded as control group in this study. Each group was assigned randomly to three tanks with 15 fish per replica. The results indicated that cold stress resulted in a significant reduction of growth metrics and a significant increase of feed conversion ratio in fish compared with MT group. Interestingly, cold stress increased hepatocyte apoptosis revealed by TUNEL staining, along with nuclear disappearance in H&E-stained sections and elevated serum alanine aminotransferase (ALT) and aspartate aminotransferase (AST) levels. Transcriptional levels of apoptosis-related genes and toll-like receptor signaling pathway components were significantly up-regulated in liver under cold stress. Compared with control group, in terms of thermal stress, the growth performance and feed utilization of fish were declined to some extent compared with MT group. Moreover, high temperature significantly elevated hepatic productions of malondialdehyde and hydrogen peroxide, as well as increased activities of some antioxidant enzymes in liver. In addition, low and high temperature induce changes in the composition of gut microbiota. Overall, the results suggested that cold stress decelerated growth performance, induced hepatocyte apoptosis, and enhanced innate immunity in hybrid sturgeon to cope with additional stressors. Whereas, thermal stress resulted in hepatic oxidative stress in liver and the protective responses in the antioxidant enzymes in fish were activated. These results provided insights into the different physiological adaptation strategies in responsive to cold stress and thermal stress in this cold-water fish.

Morphological digital assessment and transcripts of gastric and duodenal visfatin in growing piglets fed with increasing amounts of polyphenols from olive mill waste extract.

Visfatin is an adipokine with mediatory effects on inflammation. It is expressed at low levels in the pig stomach, but its role in the gastrointestinal (GI) tract is not well understood. This study explored visfatin expression and localisation in the stomach and duodenum of piglets fed varying levels of polyphenols derived from olive mill waste extract, known for their antioxidant and immunomodulatory properties. Twenty-seven piglets were assigned to three dietary groups: control (commercial feed), low polyphenol (120 ppm), and high polyphenol (240 ppm) groups. After 14 days of feeding, samples from the glandular stomach and duodenum were collected from 13 piglets. Immunohistochemistry (IHC), digital image analysis (DIA) using QuPath software, and double-labelled immunofluorescence were performed to detect visfatin-positive cells and co-localise them with serotonin. Additionally, relative gene expression of visfatin was assessed via RT-qPCR. Visfatin-positive cells were identified in 5 out of 13 piglets, localised mainly in the basal portion of gastric and intestinal glands. The morphology of those cells was consistent with neuroendocrine cells and confirmed by co-localisation of visfatin and serotonin. No significant differences were found in cell positivity or morphology between dietary groups or between tissues. However, visfatin transcript levels increased with the dose of polyphenolic extract. These findings suggest that dietary polyphenols may modulate visfatin gene expression in the GI tract. The study also highlights the value of digital anatomy for enhancing the accuracy and reproducibility of anatomical research. Further studies are needed to elucidate the functional role of visfatin transcript and protein in the porcine GI tract.

Functional characterization of β-adrenergic-like octopamine receptors in planthopper reproduction and feeding.

As an essential biogenic amine in invertebrates, octopamine (OA) regulates multiple physiological and behavioral processes via binding to octopamine receptors (OARs). The small brown planthopper Laodelphax striatellus is an important agricultural pest. However, little is known about OARs in L. striatellus. Herein, three β-adrenergic-like OARs (LsOA2s) were cloned, including LsOA2B1, LsOA2B2, and LsOA2B3. LsOA2s were expressed at the highest levels in egg stage and brain. Knockdown of LsOA2s significantly decreased the transcript levels of vitellogenin (LsVg) and Vg receptor (LsVgR), and reduced LsVg protein levels. LsOA2B2 knockdown shortened the oviposition period and inhibited the fecundity, while silencing LsOA2B1 and LsOA2B3 did not affect the reproduction performance including the preoviposition period, oviposition period, and fecundity. Ovary dissection indicated that LsOA2B2 knockdown decreased the ovary area and detained eggs. In addition, LsOA2s silencing prolonged the hatching period and reduced the hatching rate, and shortened the egg length on the fifth day of development. LsOA2B2 silencing also reduced egg width. Furthermore, LsOA2s knockdown decreased honeydew excretions of adults. Overall, these results provide evidence that LsOA2s play important roles in the reproduction and feeding behaviors of L. striatellus, and offer a reference for the exploration of potential molecular targets to control planthoppers.

Molecular responses in the intestine of Atlantic salmon (Salmo salar) following light and diet stimulation of smoltification: Potential molecular markers for a seawater-ready smolt

The transfer to seawater (SW) represents a critical stage in the production of Atlantic salmon. The success of the transfer links with the optimal development of hypo-osmoregulatory capacities during smoltification. While various strategies are adopted in aquaculture to stimulate smoltification, considerable fish loss still occurs after transfer to sea cages. Therefore, we investigated the molecular responses in the anterior and posterior intestine of Atlantic salmon, following 1) a photoperiod treatment (24 h light (L):0 h dark (D) → 24 L:0D vs. 7 L:17D → 24 L:0D) and 2) dietary treatment (regular feed or feed enriched with a salt mix/tryptophan), combined with, or without a photoperiodic treatment in freshwater (FW), to evaluate how intestinal osmoregulatory mechanisms are modulated by these treatments, and to identify potential intestinal markers indicative of a SW-ready smolt. Using quantitative real-time PCR (qPCR), we investigated transcript levels of transporters and channels involved in ion movements through the enterocytes, tight junction components, and receptors (i.e., calcium-sensing receptor and prolactin receptor). The two intestinal regions showed different gene profiles and responsiveness towards the experimental treatments. In the anterior intestine, the exposure to short photoperiod (7 L:17D) upregulated Na+/K+ − ATPase subunit alpha 1c (nkaα1c), Na+/K+/2Cl− cotransporter 1 (nkcc1), Na+/K+/2Cl− cotransporter 2 (nkcc2), Cl−/HCO3− exchanger Slc26a6 (slc26a6), and cystic fibrosis transmembrane conductance regulator I (cftrI), in FW and SW. Also, Na+/K+ − ATPase subunit alpha 1b (nkaα1b), occludin (ocln), and prolactin receptor (prlr) were upregulated in FW and claudin 15 (cldn15) in SW groups exposed to this photoperiod. The posterior intestine was less responsive to the experimental treatments, although upregulation of nkcc1, nkcc2, slc26a6, and cftrI was observed in FW in the short photoperiod groups. Hence, our findings show that exposure to a winter signal in FW more effectively activates hypo-osmoregulatory mechanisms in the intestine of Atlantic salmon, where a coordinated and complementary role of the anterior and posterior intestine ensures optimal SW processing. Dietary treatment had a positive but more marginal effect on the regulation of the genes investigated, mainly enhancing the impact of short photoperiod when the two treatments were combined. Overall, we propose the apical Na+/K+/2Cl− cotransporter, nkcc2, and the apical Cl−/HCO3− exchanger, slc26a6, as potential FW molecular markers in the anterior intestine to assess “SW-readiness” in Atlantic salmon smolts.

Functional characterization and mechanism of the multidrug resistance transport potein YoeA in Bacillus subtilis.

Transport proteins are essential for bacterial resistance to antibiotics and toxins, but their mechanisms remain poorly understood in Bacillus subtilis. In the present study, overexpression of yoeA enhanced resistance to various antibiotics, with its expression induced by these antibiotics, especially penicillin and plipastatin. The ΔyoeA strain exhibited significant growth inhibition at 100μg/mL of plipastatin, while as high as 10,000μg/mL of iturin/surfactin are required to achieve comparable inhibition, suggesting a higher sensitivity of ΔyoeA to plipastatin. The transcript level of yoeA gene was increased 2.71-fold in response to plipastatin, significantly higher than the levels induced by surfactin and iturin. The ethidium bromide (EtBr) efflux activity of YoeA was inhibited by carbonyl cyanide chlorophenylhydrazone (CCCP) and enhanced by Na+. Molecular modeling studies revealed that cation-π interactions of Na+ with Y287 and Y434 residues in the C-terminal domain of YoeA contribute to its ion channel function, and Cu2+ can form coordination bonds with the N atoms of H278 and H421 residues on the C-terminal surface of YoeA, promoting plipastatin efflux in a dose-dependent manner. The present study characterized the main factors influencing YoeA's efflux activity, revealed its transport mechanism, and provided new insights into enhancing antimicrobial peptide production and controlling bacterial resistance.

A vaccine combining ORF132 and ORF25 expressed by Saccharomyces cerevisiae induces protective immunity in Carassius auratus gibelio against CyHV-2.

CyHV-2 is the pathogen of herpesvirus haematopoietic necrosis (HVHN), resulting in significant economic losses in crucian carp. Although multiple oral vaccines have been developed to prevent CyHV-2, they have not achieved ideal protective effects. To improve the protective effect of oral vaccine, a combination vaccine was conducted by mixing recombinant Saccharomyces cerevisiae displaying ORF132 or ORF25 on the cell surface in a 1:1 ratio. Oral immunization with this combined vaccine induced specific antibodies against ORF132 and ORF25 in fish serum, as well as increased the transcription levels of IgT, IgM, IL-1β, IFN-1, and TNF-α in the spleen, head-kidney, and hindgut. Importantly, the combined vaccine provided significant protection against CyHV-2 infection in crucian carp, resulting in a relative percent survival of 80.77%. Additionally, it suppressed viral load and alleviated pathological damage of CyHV-2-infected fish. The results suggest that the combined vaccine exhibits a better protective effect compared to the single-antigen vaccines, indicating it as a promising approach to prevent CyHV-2 outbreaks.

OsIAA7 enhances heat stress tolerance by inhibiting the activity of OsARF6 in rice.

Heat stress (HS) severely affects the growth and yield of rice, necessitating a clear understanding of the molecular mechanisms underlying HS tolerance. In this study, we report that the Aux/IAA family gene, OsIAA7, whose expression is induced by HS and positively regulates HS tolerance in rice (Oryza sativa L.). The osiaa7 mutant exhibits reduced HS tolerance, whereas overexpression of OsIAA7 enhances it. Our findings suggest that OsIAA7 contributes to HS tolerance by reducing hydrogen peroxide accumulation and cell death. Physiological analysis indicates that OsIAA7 influences the levels of malondialdehyde, catalase, and chlorophyll A concentration in plants under HS conditions. RNA-seq analysis suggests that OsIAA7 modulates the expression of heat-responsive genes, contributing to HS tolerance. Further, biochemical analyses demonstrate a physical interaction between OsIAA7 and OsARF6, with OsIAA7 inhibiting the activity of OsARF6. RT-qPCR results support the notion that the positive regulatory factor OsIAA7 and the negative regulatory factor OsARF6 control HS tolerance by regulating the transcript levels of OsTT1 and OsTT3.1. Together, our results reveal the role of OsIAA7 in controlling HS tolerance through the modulation of physiological processes and the inhibition of OsARF6 activity, suggesting that some Aux/IAA family genes play a role in heat tolerance in rice.

Allatotropin, DH31, and Proctolin reduce chill tolerance in the two-spotted cricket, Gryllus bimaculatus

The ability of insects to tolerate low temperatures, known as chill tolerance, contributes to their global distribution. However, the mechanisms underlying insect chill tolerance remain poorly understood. At low temperatures, insects enter chill coma, a reversible state of paralysis, owing to disrupted ion and water homeostasis. Upon returning to normal temperatures, insects reestablish ion and water homeostasis and recover the ability to move. In this study, we used the two-spotted cricket, Gryllus bimaculatus, as an experimental model and unveiled the roles of neuropeptides in regulating chill tolerance, typically evaluated by the time taken to recover from chill coma. Screening of 37 neuropeptides revealed that Allatotropin, DH31, and Proctolin inhibited chill coma recovery and decreased the survival rate under cold stress. RT-qPCR analyses revealed that the receptors for Allatotropin and DH31 were predominantly expressed in the hindgut. Injection of the three neuropeptides decreased both hemolymph mass and gut water content at low temperatures, most likely by increasing water excretion from the hindgut due to their effects on the rectum contraction. Additionally, Allatotropin and DH31 were produced by the terminal abdominal ganglion (TAG) innervating the hindgut since they were partly co-localized in the TAG, and their mature peptides were detected in the TAG-hindgut nerves. Moreover, the transcriptional levels of the neuropeptides in the TAG and receptors in the hindgut changed with cold exposure and rewarming. Based on these findings, we propose that Allatotropin, DH31, and Proctolin affect the physiological activities of the gut, probably the hindgut, to disrupt water homeostasis at low temperatures, thereby reducing chill tolerance in crickets.

Curcumin alleviates the aggressiveness of breast cancer through inhibiting cell adhesion mediated by TEAD4-fibronectin axis

Breast cancer (BC) ranks first among women in the world and metastasis is the main reason of cancer death. TEA domain transcription factor 4 (TEAD4) plays an important role in TEAD family members in the Hippo signaling pathway, and its pro-cancer effect is gradually being discovered, but little is known about the mechanism of TEAD4 regulating tumor adhesion and metastasis. Curcumin is an active polyphenol compound with anti-inflammatory and anti-tumor properties. However, its effects and the underlying mechanisms on BC metastasis remain unknown. Here, we show that curcumin reduced the abilities of migration and invasion of BC cells as well as lung metastasis of BC in nude mice by TEAD4 induced. TEAD4 could regulate the transcription level of adhesion molecule Fibronectin (FN1), which is an important component of extracellular matrix participating in tumor cell adhesion and migration processes, and the binding of TEAD4 to the FN1 promoter was suppressed by curcumin. Furthermore, the metastatic mobility was significantly reduced in FN1 knockout BC cells, and FN1 knockout can reverse metastatic potential of TEAD4 overexpressed cells. Our work demonstrates that TEAD4 could promote cell adhesion and migration by binding with FN1 promoter and suggested that TEAD4-FN1 axis in tumor microenvironment is expected to become a potential target for alleviating metastasis of BC of curcumin.Abbreviations: Breast cancer (BC); Bicinchoninic acid (BCA); Bovine Serum Albumin (BSA); Curcumin at 20 μM (Cur20) and 30 μM (Cur30); Dimethyl −sulfoxide (DMSO); Dulbecco’s Modified Eagle’s Medium (DMEM); Extracellular Matrix (ECM); Fetal bovine serum (FBS); Fibronectin (FN1); Immunohistochemical (IHC); Minimum Essential Medium (MEM); The negative control with infected blank lentivirus (NC); Phenylmethanesulfonyl fluoride (PMSF); Polyvinylidene difluoride (PVDF); Real-time quantitative reverse transcription (RT-qPCR); single guide RNA (sgRNA); short hairpin RNA (shRNA); Human TEAD4 shRNA (shTEAD4); Transcriptional coactivator with PDZ-binding motif (TAZ); TEA domain transcription factor 4 (TEAD4); TEAD4 overexpressed (TEAD4-OE); Vascular endothelial growth factor (VEGF); Yes-associated protein (YAP);

Anticancer and therapeutic efficacy of XPO1 inhibition in pancreatic ductal adenocarcinoma through DNA damage and modulation of miR-193b/KRAS/LAMC2/ERK/AKT signaling cascade.

Pancreatic ductal adenocarcinoma (PDAC) is one of the most aggressive and grave malignancies with confined and ineffective therapeutic options. XPO1 is a critical regulator of nuclear export and activation of tumor suppressor proteins. The present study evaluated the therapeutic potential and molecular mechanisms of XPO1 inhibition against PDAC. Firstly, we observed significant overexpression of XPO1 transcript in 179 PDAC patients than 171 normal pancreatic tissues in TCGA transcriptomic dataset. Higher XPO1 transcript levels displayed worse overall and disease-free survival. Further, we confirmed significant upregulation of XPO1 in a panel of PDAC cells. Eltanexor treatment resulted in significant inhibition of cell viability, clonogenic growth, migration, and epithelial-mesenchymal transition (EMT), along with the induction of cell cycle arrest. Mechanistically, eltanexor modulated the expression of key proteins including p21, p27, p53, cyclin B1, cyclin D1, c-Myc, N-cadherin, vimentin, E-cadherin associated with the cell viability, growth, cell cycle and EMT. Additionally, the eltanexor treatment resulted in marked increase in expression of γH2AX, and cleaved PARP, cleaved caspase-9 leading to induction of DNA damage and apoptosis of PDAC cells, respectively. Moreover, eltanexor treatment regulated the expression of key non-coding RNAs including miR193b, DINO, MALAT-1, H19, and SOX21-AS1 linked with tumorigenesis. Our results revealed a correlation among miR193b/KRAS/LAMC2, XPO1/KRAS, and LAMC2/KRAS. The findings also revealed that eltanexor treatment rescued the expression of miR193b which acts as a sponge for LAMC2 and KRAS resulting in the suppression of AKT/ERK downstream signaling cascade in PDAC. Interestingly, the combination of eltanexor with gemcitabine showed significant anticancer activity in PDAC cells. Altogether, our findings revealed the crucial role of XPO1 in modulating the expression of oncogenic proteins, ncRNAs, and DNA damage during PDAC progression as well as identified novel therapeutic miR-193b/KRAS/LAMC2/ERK/AKT axis.

Composition analysis and mechanism of Guizhi Fuling capsule in anti-cisplatin-resistant ovarian cancer.

Cisplatin is the main chemotherapy drug for advanced ovarian cancer, but drug resistance often occurs. The aim of this study is to explore the molecular mechanism by which Guizhi Fuling capsule inhibits cisplatin resistance in ovarian cancer. First, differences in cisplatin resistance, PA2G4 gene expression, migration, and invasion in A2780 cells and A2780/DDP cells were analyzed by qRT-PCR, scratch assay, transwell, immunofluorescence, and western blotting. Then, LC-MS/MS analysis of GFC chemical composition. qRT-PCR, scratch tests, transwell, pseudopodium formation, immunofluorescence, and western blotting were used to explore the mechanism by which GFC inhibited A2780/DDP cell migration and invasion. Finally, the anti-tumor efficacy of GFC was verified by in vivo experiments. A2780/DDP cells had a greater ability to migrate and invade compared to their parents. Cell viability experiments showed that the migration and invasion ability of A278/DDP cells were significantly inhibited with the increase of GFC concentration. qRT-PCR results showed that compared with the blank control group, cisplatin group and GFC group, the transcription level of PA2G4 gene in the combination treatment group was significantly reduced. We also found that GFC combined with cisplatin inhibited the PI3K/AKT/GSK-3β signaling pathway by targeting PA2G4 gene expression, inhibited the epithelial-mesenchymal transition signaling pathway, decreased cell adhesion and inhibited the formation of cell pseudopodias. GFC combined with cisplatin can target PA2G4 gene to regulate PI3K/AKT/GSK-3β Signaling pathway, inhibiting the invasion and migration of cisplatin resistant A2780/DDP cells in ovarian cancer.

Boosting 2G bioethanol production by overexpressing dehydrogenase genes in Klebsiella sp. SWET4adh1+adhE: ANN optimization, scale-up, and economic sustainability

The most significant obstacle in the production of 2G bioethanol lies in transforming the complex carbohydrates found in plant biomass into simpler, fermentable sugars. However, this challenge can be effectively addressed by developing highly efficient lignocellulolytic ethanologenic strains. The current study intended to improve the ethanol productivity of efficient cellulose and hemicellulose metabolizing Klebsiella sp. SWET4 by overexpressing its native ethanologenic genes (adh1, adhE, and nifJ). Among the recombinants, SWET4adh1+adhE exhibited the highest ethanol productivity from banana peel (BP) in facultative anaerobic conditions with a 7.76-fold increase compared to the native strain. Transcript level expression study using qPCR indicated 106.15- and 22.78-fold higher expression of adh1 and adhE gene, respectively at 56 h from BP, whereas, 4.80- and 2.08-fold higher concentrations of respective proteins were obtained after 24 h. ANN and GA-mediated optimization further enhanced biomass and ethanol production to 2.33 and 24.47 g/L, respectively. Hence the ethanol productivity by SWET4adh1+adhE was enhanced by 19.89-fold. The bioreactor kinetic study exhibited diauxic growth of SWET4adh1+adhE and achieved an impressive ethanol yield of 0.44 g/g of carbohydrates, surpassing many 2G bioethanol processes. Despite of single-product scheme, a minimum selling price of $2/kg of distillate was found to make the process economically feasible. The breakeven point of the process was found to be 30% of its total capacity. Techno-economic analysis underscores the feasibility and economic advantages of the current study eliminating the pretreatment steps, and highlighting the process’s innovation and viability in 2G bioethanol production. Elimination of the pretreatment process not only achieved economic sustainability but also significantly enhanced the renewability of bioethanol production endorsing direct biomass conversion to bioethanol.

Effects of exopolysaccharides form Lactobacillus plantarum KX041 on high fat diet-induced gut microbiota and inflammatory obesity.

Lactobacillus plantarum KX041 is a probiotics obtained from early screening, which can relieve inflammation and enrich the anti-obesity intestinal flora, and produce high yield of exopolysaccharides (EPS). The extraction, structure and physicochemical properties of EPS have been completed earlier. However, whether the functional activity of L. plantarum KX041 is related to the EPS is still unclear. In this study, combined in vivo and in vitro tests explored the effects of EPS on inflammatory obesity. In vitro tests, EPS relieased inflammation by inhibiting TLR4/p-NFκB, and inhibited adipocyte differentiation and reduced the transcriptional level of inflammation and lipid-synthesis genes (especially CD36 and peroxisome proliferators-activated receptors-γ PPARγ). In vivo tests demonstrated that EPS could alleviate weight gain, white adipocyte expansion, organ damage, insulin resistance, hyperlipidemia, and inflammatory response caused by high fat diet. Combined with in vitro tests, the transcription level of inflammation and lipid-synthesis genes in adipose tissue identified that IL-1β and PPARγ were the key genes for the effect of EPS on inflammatory obesity. Moreover, EPS increased the abundance of Akkermansia and Lachnospiraceae_NK4A136_group, and intestinal levels of isobutyrate, which is one of the key factors in alleviating inflammatory obesity. These insights may inform the development of functionalized dietary interventions in obesity.