Biological Characteristics Research Articles

Evaluation of Cytotoxic Effects of Purified Mercury in Human Gingival Fibroblasts—In vitro Study

ABSTRACT Background: Since the introduction of amalgam for tooth fillings, there have been concerns that mercury toxicity could pose unacceptable health risks. Rasa shastra is an ancient medical discipline that focuses on the utilization of metals and minerals for the treatment of diseases. Nevertheless, these minerals cannot be directly administered to the human body in their natural state due to their potential adverse effects. Hence, for medicinal purposes, these metals and minerals need to undergo purification (Shodhana) to eliminate impurities and modify their physical, chemical, and biological characteristics. Methodology: Human gingival fibroblasts (HGF) were exposed to commercially available mercury (CA-Hg) and ayurvedically purified mercury (AP-Hg) at concentrations of 6.25 μM, 12.5 μM, 25 μM and 50 μM. The unexposed HGF cultured in basal media was considered a control. All the samples were cultured for 24 hours and 48 hours, and the cytotoxicity was analyzed by MTT assay. Results: Cell viability between the control and experimental groups varied at 24 hours, however, the results were not statistically significant (p>0.05). At 48 hours, cell viability was higher in the AP-Hg group as compared to the CA-Hg group at the concentration of 6.25 μM, and the difference was statistically significant (p<0.05). The cell proliferation assay results demonstrated a statistically significant difference in the mean optical density values (p<0.05) between CA-Hg and AP-Hg at 12.50 μM, 25 μM, and 50, μM concentrations observed at 24 hours. At 48 hours, a statistically significant difference in the mean OD values (p<0.05) between CA-Hg and AP-Hg at all four concentrations was observed. Conclusion: AP-Hg at a concentration of 6.25 μM demonstrated higher cell viability at 48 hours. Further, the cell proliferation rate was also higher for AP-Hg at all concentrations at 24 and 48 hours. These results indicated a less cytotoxic effect of AP-Hg than CA-Hg in HGF and hence could be employed for dental amalgam preparations.

Spanioneura fonscolombii Förster (Hemiptera: Psylloidea) a New Jumping Plant-Lice for Turkish Fauna

Aim of study: To investigate the distribution area, some biological characteristics and host plant relationship of Spanioneura fonscolombii Förster (Hemiptera: Psylloidea) species, a psyllid we recently found its distribution in Turkey, for the first time within this study. 
 Material and methods: We collected adults and nymphs of the insect from the host plants during the visits to the study area on different dates. The insect was recognised as a psyllid species. Some adult samples were sent to the expert Dr. Burckhard for species identification. The psyllid damage on host plants of Buxus balearica was identified for the first time as a new host for Spanioneura fonscolombii. 
 Main results: 1- We identified Spaineura fonscolombii species distribution in Turkey for the first time, 2- Population size was highly dense in the study area based on initial observation, 3- New host plant, Buxus balearica, for the pest species was identified for the first time, 4- Overwintering properties of the pest species was determined. 
 Research highlihts: First record of the pest in Turkey. Determination of the most Southern and Eastern, probably so far known, distribution of the pest species in the world. Determination of a new host plant for the pest.

MTOR Inhibition Prolongs Survival and Has Beneficial Effects on Heart Function After Onset of Lamin A/C Gene Mutation Cardiomyopathy in Mice.

Mutations in LMNA encoding nuclear envelope proteins lamin A/C cause dilated cardiomyopathy. Activation of the AKT/mTOR (RAC-α serine/threonine-protein kinase/mammalian target of rapamycin) pathway is implicated as a potential pathophysiologic mechanism. The aim of this study was to assess whether pharmacological inhibition of mTOR signaling has beneficial effects on heart function and prolongs survival in a mouse model of the disease, after onset of heart failure. We treated male LmnaH222P/H222P mice, after the onset of heart failure, with placebo or either of 2 orally bioavailable mTOR inhibitors: everolimus or NV-20494, a rapamycin analog highly selective against mTORC1. We examined left ventricular remodeling, and the cell biological, biochemical, and histopathologic features of cardiomyopathy, potential drug toxicity, and survival. Everolimus treatment (n=17) significantly reduced left ventricular dilatation and increased contractility on echocardiography, with a 7% (P=0.018) reduction in left ventricular end-diastolic diameter and a 39% (P=0.0159) increase fractional shortening compared with placebo (n=17) after 6 weeks of treatment. NV-20494 treatment (n=15) yielded similar but more modest and nonsignificant changes. Neither drug prevented the development of cardiac fibrosis. Drug treatment reactivated suppressed autophagy and inhibited mTORC1 signaling in the heart, although everolimus was more potent. With regards to drug toxicity, everolimus alone led to a modest degree of glucose intolerance during glucose challenge. Everolimus (n=20) and NV-20494 (n=20) significantly prolonged median survival in LmnaH222P/H222P mice, by 9% (P=0.0348) and 11% (P=0.0206), respectively, compared with placebo (n=20). These results suggest that mTOR inhibitors may be beneficial in patients with cardiomyopathy caused by LMNA mutations and that further study is warranted.

Response of bacterial communities, enzyme activities and dynamic changes of soil organic nitrogen fractions to six-year different application levels of biochar retention in Northeast China

Composition and availability of soil organic nitrogen play a key role in soil N retention and supply. The application of traditional chemical fertilizers can improve soil production and crop yields. However, it also leads to low N utilization efficiency and soil solidification. Previous studies have shown that fertilizers added with biochar can improve soil fertility and facilitate soil nutrient transformation by regulating soil biological indicators. However, whether biochar used with fertilizers can promote nutrient cycling in the long term, which remains to be tested. Therefore, this study aims to explore the long-term effects of biochar with different levels used with fertilizers on soil fertility, N supply potential of organic N, and microbial communities, thus, providing a theoretical reference for the rational management of biochar applications. In this study, a field experiment was carried out from 2013 to 2018 assess dynamic changes in organic N fractions and diversity and richness of microbial communities under the impacts of chemical fertilizers combined with different levels of biochar. Biochar with no amendment (CK), biochar with 15.75 t ha−1 (BC1), biochar with 31.5 t ha−1 (BC2), and biochar with 47.25 t ha−1 (BC3) are four biochar levels that were tested in this study. The results show that compared with CK, those other three biochar levels can increase soil organic matter, C/N, and pH values. Acid-hydrolysable N (AHN) makes a major contribution to organic N in soil. Biochar retention for six years has significantly affected AHN levels, as well as its four fractions, namely, amino acid N (AAN), acid-hydrolysable ammonium N (AN), amino sugar N (ASN), and hydrolysable unknown N (HUN). Some vital biological characteristics, with which each AHN fraction is primarily regulated, can be altered with the application of biochar. That is, AAN is mainly controlled by enriched Verrucomicrobia, Cyanobacteria, Glucosylbacteria, and Nitrospira, and reduced soil urease, while AN is mainly controlled by enriched Verrucomicrobia, Bacteroidetes, Gemmatimonadetes, Acidobacteria, and Proteobacteria. ASN is regulated by enriched Acidobacteria and depleted soil protease, while HUN is regulated by Gemmatimonadetes and enriched soil nitrate reductase. It has been found that microbial and soil enzyme activities are involved in N cycle processes, indicating that the formation of AHN fractions is closely associated with biochar-induced specific microbial and enzyme activities. This study has provided new insights into the influences of biochar with different levels on distribution of soil organic N fractions, and shifts in soil enzyme activities and microbial communities.

A Review on Effect of Organic Conditioner on Physico-chemical and Microbiological Properties of Soil

The unbalanced use of chemical fertilizers has caused adverse effects on the environment and human health, which has raised the focus on environmentally friendly agricultural techniques for sustainable farming globally. The article investigates the effects of organic conditioners on the physical, chemical, and microbiological characteristics of soil, with focus on liquid organic conditioners such Panchagavya, Jeevamruth, and Beejamruth. In order to purify agricultural fields, the necessity of organic agriculture methods that are both commercially and environmentally feasible is emphasized. The advantages of applying Soil conditioners are explained, including increased crop production, quality, affordability, and safety for the environment in addition to increases in soil health, fertility, and pest resistance. An overview of the basic principles of liquid organic manures is provided, with a focus on their ability to release nutrients, stimulate microbial activity, and facilitate plant absorption. The review discusses how organic conditioners might improve the biological characteristics of soil, such as the presence of chemicals that promote plant growth and helpful microbes. Liquid organic manure adoption is essential for healthy ecosystems, sustainable agriculture, and increased soil fertility and productivity. Because liquid organic manures may be applied in a variety of ways and used as soil drenches, foliar sprays, and seedling treatments, they are a crucial part of environmentally friendly farming methods. It is advised to combine liquid and solid organic manures to reduce negative effects and guarantee a robust and fruitful farming environment.

Discovery of a Novel Pseudo-Natural Product Aurora Kinase Inhibitor Chemotype through Morphological Profiling.

The pseudo-natural product (pseudo-NP) concept aims to combine NP fragments in arrangements that are not accessible through known biosynthetic pathways. The resulting compounds retain the biological relevance of NPs but are not yet linked to bioactivities and may therefore be best evaluated by unbiased screening methods resulting in the identification of unexpected or unprecedented bioactivities. Herein, various NP fragments are combined with a tricyclic core connectivity via interrupted Fischer indole and indole dearomatization reactions to provide a collection of highly three-dimensional pseudo-NPs. Target hypothesis generation by morphological profiling via the cell painting assay guides the identification of an unprecedented chemotype for Aurora kinase inhibition with both its relatively highly 3D structure and its physicochemical properties being very different from known inhibitors. Biochemical and cell biological characterization indicate that the phenotype identified by the cell painting assay corresponds to the inhibition of Aurora kinase B.

Correlation of Soil Chemical, Biological, and Physical Characteristics on Productivity of Maize in Cibugel District, Sumedang Regency

Maize as one of the national food commodities, experiences an increase in demand every year. Both internal and external factors influenced the growth of corn plants. Internal factors originated from the plant, such as inherited genetic traits, while external factors came from the environment, including soil properties and climate. External factors that affected plant growth included biological, physical, and chemical soil characteristics. This research aimed to understand the interrelation between soil acidity (pH), phosphorus availability (P), cation exchange capacity (CEC), nitrogen-fixing bacteria, earthworm population, carbon-nitrogen ratio (C/N), and soil texture on the productivity of maize in Cibugel District and how these factors were interconnected to support plant growth. The research employed a descriptive survey and comparative method, with soil samples taken from 18 soil samples from six villages in the Cibugel Subdistrict, Sumedang Regency. The correlation analysis results showed a significant positive relationship between cation exchange capacity and corn productivity and a significant negative relationship between available phosphorus and corn productivity. This implied that increasing cation exchange capacity could enhance productivity while increasing available phosphorus could decrease productivity. The research aided in identifying soil characteristics influencing high land productivity in the area. 

Conserved methylation signatures associate with the tumor immune microenvironment and immunotherapy response

BackgroundAberrant DNA methylation is a major characteristic of cancer genomes. It remains unclear which biological processes determine epigenetic reprogramming and how these processes influence the variants in the cancer methylome, which can further impact cancer phenotypes.MethodsWe performed pairwise permutations of 381,900 loci in 569 paired DNA methylation profiles of cancer tissue and matched normal tissue from The Cancer Genome Atlas (TCGA) and defined conserved differentially methylated positions (DMPs) based on the resulting null distribution. Then, we derived independent methylation signatures from 2,465 cancer-only methylation profiles from the TCGA and 241 cell line-based methylation profiles from the Genomics of Drug Sensitivity in Cancer (GDSC) cohort using nonnegative matrix factorization (NMF). We correlated DNA methylation signatures with various clinical and biological features, including age, survival, cancer stage, tumor immune microenvironment factors, and immunotherapy response. We inferred the determinant genes of these methylation signatures by integrating genomic and transcriptomic data and evaluated the impact of these signatures on cancer phenotypes in independent bulk and single-cell RNA/methylome cohorts.ResultsWe identified 7,364 differentially methylated positions (2,969 Hyper-DMPs and 4,395 Hypo-DMPs) in nine cancer types from the TCGA. We subsequently retrieved three highly conserved, independent methylation signatures (Hyper-MS1, Hypo-MS1, and Hypo-MS4) from cancer tissues and cell lines based on these Hyper and Hypo-DMPs. Our data suggested that Hypo-MS4 activity predicts poor survival and is associated with immunotherapy response and distant tumor metastasis, and Hypo-MS4 activity is related to TP53 mutation and FOXA1 binding specificity. In addition, we demonstrated a correlation between the activities of Hypo-MS4 in cancer cells and the fractions of regulatory CD4 + T cells with the expression levels of immunological genes in the tumor immune microenvironment.ConclusionsOur findings demonstrated that the methylation signatures of distinct biological processes are associated with immune activity in the cancer microenvironment and predict immunotherapy response.

Spatio-temporal connectivity and dispersal seasonal patterns in the Adriatic Sea using a retention clock approach

Hydrodynamic features play a key role in determining the dispersal and connectivity of fish populations, especially in highly energetic areas determined by currents, river flow, and meteorologically induced fluctuations. Understanding how species interact with these physical processes is essential for managing vulnerable populations and identifying areas that require effective conservation efforts. This study examines the hydrodynamics that regulate connectivity in the Adriatic Sea, a shallow and semi-enclosed basin that is widely recognized as one of the most important areas in the Mediterranean Sea for protection. A high-resolution hydrodynamic model coupled with a lagrangian tracking module serves as the numerical tool. Lagrangian particles, representing eggs and larvae with typical biological characteristics of generic marine organisms inhabiting the region, are released throughout the basin at different times during a test year to identify the most likely pathways of individual dispersal. The temporal component of connectivity is highlighted using a previously developed retention clock matrix over different larval durations. Seasonality is a critical factor in dispersal, with greater variability and reduced efficiency in winter compared to summer. The potential implications of the results for improved assessment and management of high value marine species in the basin are discussed.

Computational limits to the legibility of the imaged human brain

Our knowledge of the organisation of the human brain at the population-level is yet to translate into power to predict functional differences at the individual-level, limiting clinical applications and casting doubt on the generalisability of inferred mechanisms. It remains unknown whether the difficulty arises from the absence of individuating biological patterns within the brain, or from limited power to access them with the models and compute at our disposal.Here we comprehensively investigate the resolvability of such patterns with data and compute at unprecedented scale. Across 23 810 unique participants from UK Biobank, we systematically evaluate the predictability of 25 individual biological characteristics, from all available combinations of structural and functional neuroimaging data. Over 4526 GPU*hours of computation, we train, optimize, and evaluate out-of-sample 700 individual predictive models, including fully-connected feed-forward neural networks of demographic, psychological, serological, chronic disease, and functional connectivity characteristics, and both uni- and multi-modal 3D convolutional neural network models of macro- and micro-structural brain imaging.We find a marked discrepancy between the high predictability of sex (balanced accuracy 99.7%), age (mean absolute error 2.048 years, R2 0.859), and weight (mean absolute error 2.609Kg, R2 0.625), for which we set new state-of-the-art performance, and the surprisingly low predictability of other characteristics. Neither structural nor functional imaging predicted an individual's psychology better than the coincidence of common chronic disease (p < 0.05). Serology predicted chronic disease (p < 0.05) and was best predicted by it (p < 0.001), followed by structural neuroimaging (p < 0.05).Our findings suggest either more informative imaging or more powerful models will be needed to decipher individual level characteristics from the human brain. We make our models and code openly available.

Concomitant investigation of crustacean amphipods lipidome and metabolome during the molting cycle by Zeno SWATH data-independent acquisition coupled with electron activated dissociation and machine learning

BackgroundDIA (Data-Independent Acquisition) is a powerful technique in Liquid Chromatography coupled with high-resolution tandem Mass Spectrometry (LC-MS/MS) initially developed for proteomics studies and recently emerging in metabolomics and lipidomics. It provides a comprehensive and unbiased coverage of molecules with improved reproducibility and quantitative accuracy compared to Data-Dependent Acquisition (DDA). Combined with the Zeno trap and Electron-Activated Dissociation (EAD), DIA enhances data quality and structural elucidation compared to conventional fragmentation under CID. These tools were applied to study the lipidome and metabolome of the freshwater amphipod Gammarus fossarum, successfully discriminating stages and highlighting significant biological features. Despite being underused, DIA, along with the Zeno trap and EAD, holds great potential for advancing research in the omics field. ResultsDIA combined with the Zeno trap enhances detection reproducibility compared to conventional DDA, improving fragmentation spectra quality and putative identifications. LC coupled with Zeno-SWATH-DIA methods were used to characterize molecular changes in reproductive cycle of female gammarids. Multivariate data analysis including Principal Component Analysis and Partial Least Square Discriminant Analysis successfully identified significant features. EAD fragmentation helped to identify unknown features and to confirm their molecular structure using fragmentation spectra database annotation or machine learning. EAD database matching accurately annotated five glycerophospholipids, including the position of double bonds on fatty acid chain moieties. SIRIUS database predicted structures of unknown features based on experimental fragmentation spectra to compensate for database incompleteness. SignificanceReproducible detection of features and confident identification of putative compounds are pivotal stages within analytical pipelines. The DIA approach combined with Zeno pulsing enhances detection sensitivity and targeted fragmentation with EAD in positive polarity provides orthogonal fragmentation information. In our study, Zeno-DIA and EAD thereby facilitated a comprehensive and insightful exploration of pertinent biological molecules associated with the reproductive cycle of gammarids. The developed methodology holds great promises for identifying informative biomarkers on the health status of an environmental sentinel species.

The Role of Interleukin 23/17 Axis in Psoriasis Management: A Comprehensive Review of Clinical Trials.

Psoriasis pathogenesis is influenced by genetic factors and characterized by a complex interplay between genetic predisposition and various environmental triggers. These triggers set off metabolic processes involving inflammation, cell signaling, immune response dysregulation, and antigen presentation. Several types of innate and adaptive immune cells are involved in psoriasis. Among the cytokine cascade which leads to psoriasis development, the interleukin (IL)-23/Th17 axis, especially IL-17 production, emerges as crucial. Recognizing the pivotal role of this axis has facilitated the development of selective and effective biological drugs, such as anti-IL17 and anti-IL23 monoclonal antibodies. These drugs aim to achieve the complete or near-complete disappearance of psoriatic lesions, as indicated by PASI100 and PASI90 responses, respectively. In this context, the aim of our review was to delve into the functioning of the IL-23/Th17 axis, its dysregulation in psoriasis pathogenesis, and the therapeutic potential of its inhibition. Currently, 4 anti-IL17 (secukinumab, ixekizumab, bimekizumab and brodalumab) and 3 anti-IL23 (guselkumab, risankizumab and tildrakizumab) have been approved. All these drugs showed high levels of effectiveness in both clinical trials and real-life experiences, with an excellent profile in terms of safety. Certainly, furthers studies will allow for better characterization of biologics' profile, in order to administer the right drug for the right patients at the right moment.

The role of immuno-metabolic depression features in the effects of light therapy in patients with depression and type 2 diabetes mellitus: A randomized controlled trial

ObjectiveImmuno-metabolic depression (IMD) is proposed to be a form of depression encompassing atypical, energy-related symptoms (AES), low-grade inflammation and metabolic dysregulations. Light therapy may alleviate AES by modulating inflammatory and metabolic pathways. We investigated whether light therapy improves clinical and biological IMD features and whether effects of light therapy on AES or depressive symptom severity are moderated by baseline IMD features. Associations between changes in symptoms and biomarkers were explored. MethodsIn secondary analyses, clinical trial data was used from 77 individuals with depression and type 2 diabetes mellitus (T2DM) randomized to four weeks of light therapy or placebo. AES severity and depressive symptom severity were based on the Inventory of Depressive Symptomatology. Biomarkers included 73 metabolites (Nightingale) summarized in three principal components and CRP, IL-6, TNF-α, INF-γ. Linear regression analyses were performed. ResultsLight therapy had no effect on AES severity, inflammatory markers and metabolite principle components versus placebo. None of these baseline features moderated the effects of light therapy on AES severity. Only a principle component reflecting metabolites implicated in glucose homeostasis moderated the effects of light therapy on depressive symptom severity (βinteraction = 0.65, P = 0.001, FDR = 0.003). Changes in AES were not associated with changes in biomarkers. ConclusionFindings do not support the efficacy of light therapy in reducing IMD features in patients with depression and T2DM. We find limited evidence that light therapy is a more beneficial depression treatment among those with more IMD features. Changes in clinical and biological IMD features did not align over four-weeks' time.Trial registration: The Netherlands Trial Register (NTR) NTR4942.

Unraveling the Key step in the Aroma Puzzle: Insights into Alcohol Acyltransferases in Strawberries

Alcohol acyltransferases (AATs) play a crucial role in catalyzing the transfer of acyl groups, contributing to the diverse aroma of fruits, including strawberries. In this research we identified nine AAT genes in strawberries through a comprehensive analysis involving phylogenetics, gene structure, conserved motifs, and structural protein model examinations. The study used the 'Camarosa' strawberry genome database, and experiments were conducted with fruits harvested at different developmental and ripening stages. The transcriptional analysis revealed differential expression patterns among the AAT genes during fruit ripening, with only four genes (SAAT, FaAAT2, FaAAT7, and FaAAT9) showing increased transcript accumulation correlated with total AAT enzyme activity. Additionally, the study employed in silico methods, including sequence alignment, phylogenetic analysis, and structural modeling, to gain insights into the AAT protein model structures with increase expression pattern during fruit ripening. The four modeled AAT proteins exhibited structural similarities, including conserved catalytic sites and solvent channels. Furthermore, the research investigated the interaction of AAT proteins with different substrates, highlighting the enzymes' promiscuity in substrate preferences. The study contributes with valuable information to unveil AAT gene family members in strawberries, providing scientific background for further exploration of their biological characteristics and their role in aroma biosynthesis during fruit ripening.

Analyzing Pooled Microarray Gene Expression Data to Uncover Common Pathways in Periodontitis and Oral Squamous Cell Carcinoma from the Gene Expression Omnibus

ABSTRACT Periodontitis and oral squamous cell carcinoma (OSCC) are prevalent oral diseases with distinct etiologies, yet they share certain molecular and biological characteristics. Gene expression datasets from the gene expression omnibus (GEO) repository (GSE30784 for OSCC and GSE10334 for periodontitis) were analyzed. Data preprocessing and differential gene expression analysis using GEO2R identified common differentially expressed genes (DEGs), and FunRich software facilitated the construction of a protein-protein interaction (PPI) network on the STRING database. Cytoscape, coupled with the CytoHubba plugin, identified Cluster of Differentiation 19 (CD19) and Von Willebrand Factor (VWF) as the top hub genes, with Complement C3 (C3) also highly ranked. Functional enrichment analysis highlighted pathways such as the B-cell receptor signaling pathway, complement and coagulation cascades, and hematopoietic cell lineage. Additionally, miRNet analysis identified key miRNAs, including hsa-mir-26a-5p, hsa-mir-129-2-3p, and hsa-mir-27a-3p, associated with these pathways. These findings suggested an association of molecular mechanisms between periodontitis and OSCC, with identified hub genes and miRNAs potentially serving as therapeutic targets.

Optically induced dielectrophoresis for continuous separation and purification of C. vulgaris and H. pluvialis

Marine microalgae are widely present in the natural environment, exhibiting a significant economic value. However, during the inoculation and cultivation process of microalgae, the introduction of unwanted algae is bound to trigger nutrient competition, leading to a decrease in the growth rate of microalgae and consequently impacting their economic value in production. To address this issue, this study integrates the optically induced dielectrophoresis (ODEP) manipulation technology based on the continuous flow in a microfluidic system. A two-stage cell filter, utilizing two virtual optical spots, is designed and manufactured. Leveraging the size differences between microalgae, continuous separation and purification of mixed samples containing Chlorella vulgaris and Haematococcus pluvialis are achieved within microchannels. Additionally, optimal ODEP manipulation conditions for mixed algal liquid samples, comprising C. vulgaris and H. pluvialis, are demonstrated, including appropriate alternating current voltage (6 V), alternating current frequency (100 kHz), light spot width (40 μm), and sample flow rate (0.9 μl/min). Analysis of mixed liquid samples collected at the chip's outlet reveals a reduction in the proportion of H. pluvialis from 37.5% to 1.2% after separation. In summary, this study proposes an ODEP microfluidic system capable of continuously separating and purifying microalgae with different biological characteristics, showcasing its potential as an alternative to traditional labor-intensive microalgae separation techniques.

Prognostic marker of immunohistochemistry-based somatostatin receptors 2 and 5 H-scores in patients with pancreatic neuroendocrine neoplasms

ObjectivePancreatic neuroendocrine neoplasms (pNENs) are histologically classified as well-differentiated, poorly-differentiated, or mixed neuroendocrine-non-neuroendocrine neoplasms. There are unresectable pNENs owing to metastases or invasion in not only functional pNENs but also non-functional. However, the exact origin of pNENs has not been elucidated. This study aims to characterize the molecular biology of pNENs based on clinical information and histopathological analysis and identify prognostic biomarkers. MethodsWe investigated the relationship between the biological characteristics and immunostaining of pathological tissues in 75 patients. Staining density was evaluated on a 4-point scale from 0 to 3, and the percentage of tumor cells was calculated and scored from 0 to 300 (H-score). We performed receiver operating characteristic (ROC) curve analysis of the H-score. Progression-free survival and overall survival analyses were performed based on the Kaplan–Meier curves. ResultsThe H-score showed that patients who died of pNEN had high Ki-67 and low somatostatin receptor (SSTR) 2 levels, and those who relapsed had high Ki-67 and low SSTR5 levels. The ROC showed that the SSTR2 H-score > 80.25 was associated with lower mortality, which was further confirmed by Kaplan–Meier curves [hazard ratio (HR): 6.039, 95 % confidence interval (CI): 1.233–29.59, P = 0.0006). SSTR5 H-score > 93.9 had less recurrence, which was confirmed using Kaplan–Meier curves (HR: 3.321, 95 % CI: 1.426–7.734, P = 0.0336). ConclusionKi-67 > 4.95 is associated with a significantly increased risk of death. Quantification of SSTR2 and SSTR5 immunostaining using the H-score may serve as prognostic markers.

[Advancements and controversies in conversion therapy for biliary tract malignant tumors with insufficient future liver remnant].

Due to the unique location and aggressive tumor biology,hilar cholangiocarcinoma,intrahepatic cholangiocarcinoma,and gallbladder cancer often present with obstructive jaundice and require extensive liver resection,also exhibit high rates of recurrence and metastasis after radical excision. Therefore,surgeons should make treatment decisions based on the biliary anatomy of patients and the biological characteristics of tumors as it significantly affects patient's prognosis. Treatment strategy should be made to ensure the successful implementation of radical resection for biliary tract malignant tumors while maximizing the survival benefits of patients. Firstly,conversion of liver function by relieving jaundice technology and conversion of tumor biological characteristics through systematic therapy,followed by the conversion of future liver remnant. Currently,there are still controversies surrounding indications,methods,standards of relieving jaundice,and treatment plans,cycles,evaluation of therapeutic effects for systematic conversion therapy,and the standards and techniques of conversion therapy for future liver remnant.This article discusses these issues through literature analysis and the author's experience in the hope of resonating with colleagues.

INHBA regulates Hippo signaling to confer 5-FU chemoresistance mediated by cellular senescence in colon cancer cells

Colon cancer has become a global public health challenge, and 5-Fluorouracil (5-FU) chemoresistance is a major obstacle in its treatment. Chemoresistance can be mediated by therapy-induced cellular senescence. This study intended to investigate mechanisms of INHBA (inhibin A) in 5-FU resistance mediated by cellular senescence in colon cancer. Bioinformatics analysis of INHBA expression in colon cancer tissues, survival analysis, and correlation analysis of cellular senescence markers were performed. The effects of INHBA on the biological characteristics and 5-FU resistance of colon cancer cells were examined through loss/gain-of-function and molecular assays. Finally, a xenograft mouse model was built to validate the mechanism of INHBA in vivo. INHBA was upregulated in colon cancer and was significantly positively correlated with cellular senescence markers uncoupling protein 2 (UCP-2), matrix metalloproteinase-1 (MMP-1), dense and erect panicle 1 (DEP1), and p21. Cellular senescence in colon cancer mediated 5-FU resistance. Downregulation of INHBA expression enhanced 5-FU sensitivity in colon cancer cells, inhibited cell proliferation, promoted apoptosis, increased the proportion of cells in G0/G1 phase, and it resulted in a lower proportion of senescent cells and lower levels of the cellular senescence markers interleukin 6 (IL-6) and interleukin 8 (IL-8). Analysis of whether to use the pathway inhibitor Verteporfin proved that INHBA facilitated colon cancer cell senescence and enhanced 5-FU chemoresistance via inactivation of Hippo signaling pathway, and consistent results were obtained in vivo. Collectively, INHBA conferred 5-FU chemoresistance mediated by cellular senescence in colon cancer cells through negative regulation of Hippo signaling.

Multifunctional Platform for Future Applications in Cell and Tissue Engineering based on Silicate Phosphate Hydroxyapatite Co-doped with Li+, Eu3+ and Gd3+ ions

Although hydroxyapatite has established itself as a promising biomaterial for the simultaneous diagnosis and treatment of bone defects, several physicochemical properties can be further refined to achieve a specialized theranostic platform for cell and tissue engineering. In this context, we synthesized silicate-substituted hydroxyapatites co-doped with Li+, Eu3+, and Gd3+ ions (abbr. as Si-HAp-LEG) using a microwave-assisted hydrothermal method. The aim was to create a combination of dopants to stimulate bone regeneration (Li+) while enabling bioimaging (Eu3+ and Gd3+). Structural and morphological assessments included, e.g., XRPD (X-ray Powder Diffraction), HR-TEM (High-resolution Transmission Electron Microscopy) imaging, and BET (Brauner-Emmet-Teller) analysis. Unit-cell parameters were estimated via Rietveld refinement. The luminescence properties (emission, excitation spectra, and emission kinetics) were recorded depending on the varying Li+ concentration. The cytocompatibility of biomaterials was tested using hBMSCs (multipotent human bone-marrow mesenchymal stromal cells). In vitro assays evaluated biomaterials’ effects on morphology, metabolic activity (Alamar blue assay), and transcriptomic activity (RT-qPCR). The results indicate that co-doping with Li+, Eu3+, and Gd3+ ions introduces distortions while preserving the crystal structure of hydroxyapatite. Additionally, the cells maintained proper morphology and ultrastructure, internalizing biomaterials without significant cytotoxicity. The biomaterials also triggered the expression of key transcriptional factors. Si-HAp-LEG biomaterials, particularly LEG-222 and LEG-221, exhibit desired physicochemical and biological features, rendering them highly bioactive and promising for cell and tissue engineering applications.