Tissue Distribution Research Articles

Improving computational models of deep brain stimulation through experimental calibration.

Deep brain stimulation has become a well-established clinical tool to treat movement disorders. Nevertheless, the knowledge of processes initiated by the stimulation remains limited. To address this knowledge gap, computational models are developed to gain deeper insight. However, their predictive power remains constrained by model uncertainties and a lack of validation and calibration. Exemplified with rodent microelectrodes, we present a workflow for validating electrode model geometry using microscopy and impedance spectroscopy in vitro before implantation. We address uncertainties in the tissue distribution and dielectric properties and outline a concept for calibrating the computational model based on in vivo impedance spectroscopy measurements. The standard deviation of the volume of tissue activated across the 18 characterized electrodes was approximately 32.93%, underscoring the importance of electrode characterization. Thus, the workflow significantly enhances the model predictions' credibility of neural activation exemplified in a rodent model. Computational models are frequently employed without validation or calibration, relying instead on manufacturers' specifications. Our approach provides an accessible method to obtain a validated and calibrated electrode geometry, which significantly enhances the reliability of the computational model that relies on this electrode. By reducing the uncertainties of the model, the accuracy in predicting neural activation is increased. The entire workflow is realized in open-source software, making it adaptable for other use cases, such as deep brain stimulation in humans. Additionally, the framework allows for the integration of further experiments, enabling live updates and refinements to computational models.

Development and optimization of multivesicular gefitinib liposomal transdermal system employing lipoid S100 for breast cancer: pharmacokinetics, bioavailability, and skin irritation studies in Wistar rats

Abstract Background Conventional therapies in cancer treatment face challenges including drug resistance, lack of specificity, and severe adverse reactions. This study explores the potential of liposomal transdermal delivery systems as an alternative to current therapies with improved BA and PK. The objective of the study was to formulate gefitinib liposomes by thin film hydration technique (TFH) using lipoid S100. A central composite design (CCD) was used to develop and optimize GEF-LIP-TDDs and to analyze the optimum concentrations of the selected variables (phospholipid, cholesterol) in liposomal formation. The model fitting was performed using Design-Expert (Stat-Ease, Ver 13). The GEF liposomes were evaluated for %EE, mean particle size and PDI. The optimized liposomes were fabricated as a transdermal patch by mercury substrate method and evaluated for %drug content, in vitro diffusion, in vivo biodistribution (PK and BA), and skin irritation studies in female Albino Wistar rats. The stability of the optimized transdermal patch was also assessed for 3 months. Results The CCD model was significant with F-value of 37.97, P-value of 0.0500 and R 2 of 0.9644. The average vesicle size, PDI, and ZP of GEF-LIPs (F1–F13) were found to be between 112.8 to 373.7 nm, 0.186 to 0.510 and − 3.69 to − 82.2 mV, respectively. F3-GEF-LIP exhibited a mean vesicle size of 96.07 nm, ZP of − 46.06 mV, and a PDI of 0.423. F3-GEF-LIP demonstrated exceptional %EE (97.79) and sustained release effect (%CDR, 83.32) following a diffusion-controlled mechanism. TEM images confirmed liposomes of multivesicular type (MVV, < 100 nm). Importantly, optimized F3-GEF-LIP-TD showed no signs of edema in Wistar rats. The biodistribution of F3-GEF-LIP-TD was similar to pure GEF and was higher in the liver (p < 0.05). The BA of F3-GEF-LIP-TD was observed to be 74.05 ± 0.11% in comparison with oral GEF-LIP (65.25 ± 0.08%) and pure GEF (58.10 ± 0.17%). Conclusion TFH technique offers stable liposomes with high reproducibility. Our findings imply that GEF-LIP-TD provides enhanced BA and tissue distribution and can be considered as a substitution for orals or in combination for treating breast cancer. Lipoid S100 is a potential lipid for developing stable multivesicular nanoliposomes. Graphical abstract

Tissue distribution of naringenin conjugated metabolites following repeated dosing of naringin to rats

Tissue distribution of naringenin conjugated metabolites following repeated dosing of naringin to rats

Preclinical delayed toxicity studies of BCMA CAR T-cell injection in B-NDG mice with multiple myeloma

PurposeBased on the efficacy data from the previous study of B-cell maturation antigen (BCMA) chimeric antigen receptor (CAR) T-cell injection, we further examined the delayed toxicity for 8 weeks after a single dose of BCMA CAR T-cell injection to observe possible toxic reactions.MethodsB-NDG mice transplanted with multiple myeloma (MM) cells were given a single dose of BCMA CAR T-cell injection at two dosages or human normal T cells and then subjected to examinations including clinical signs, weight and food intake measurements, haematology, blood biochemical analysis, cytokine assay, T-lymphocyte subpopulation quantification and histopathology on days 28 and 56 after dosing. In addition, quantitative polymerase chain reaction (qPCR) was used to quantify DNA fragments in different tissues to assess the tissue distribution of CAR and provide a basis for its preclinical safety evaluation and clinical dosing.ResultsIn the delayed toxicity study, no mortality or significant toxic effects such as reductions in food intake, body weight, relevant biochemical parameters and target organ weights were observed in the BCMA CAR T-cell-treated groups. Compared to the model group, restorative changes in clinical signs and clinicopathology indicating therapeutic effects were seen in the BCMA CAR T-cell-treated groups. Human-derived cytokines interleukin-2 (IL-2), IL-4, IL-6, IL-12, IL-10, tumor necrosis factor α (TNF-α), and interferon-γ (IFN-γ) could be detected in all cancer cell–bearing mice by cytokine level measurement. IFN-γ levels showed a geometric increase due to the graft versus host disease (GVHD) response induced in the mice, while the levels of the other cytokines did not show significant changes. Histopathological examination indicated that the BCMA CAR T-cell treatment groups showed mixed cellular infiltration of human-derived T cells, cancer cells, and inflammatory cells in several target organs including the liver, spleen, lung, and kidney, and some of them showed mild tissue damage, but the number of the animals and the severity of damage were significantly less than those of the T-cell control group as well as the model group. The results of the tissue distribution study showed that BCMA CAR T cells were mainly concentrated in the kidney, lung, bone marrow and the related immune organs/tissues, and the distribution of BCMA CAR T cells was highly consistent with that of MM cells, suggesting that BCMA CAR T cells could follow the cancer cells during metastatic targeting of the tissues.ConclusionsThe present study demonstrated a low toxicity of BCMA CAR T-cell injection, with manageable side effects and good anticancer activity and without observable adverse effects. This study provides data to support future clinical studies of BCMA CAR T-cell injection for MM.

Abstract 4136412: Quantitative Cardiac Magnetic Resonance Standardized Signal Intensity Comparison in Dilated Cardiomyopathy versus Cardiac Sarcoidosis

Introduction/Background: Dilated cardiomyopathy (DCM) and cardiac sarcoidosis (CS) manifest unique late gadolinium enhancement (LGE) patterns on cardiac magnetic resonance (CMR), indicative of differing distributions of myocardial scar tissue. Nevertheless, these LGE patterns lack specificity, leading to significant overlap between the two conditions. Goals/Aims: This study seeks to introduce a novel quantitative method employing z-score analysis of LGE-CMR signal intensity to objectively evaluate and compare the spatial distribution of LGE intensity between DCM and CS. Methods/Approach: The retrospective cohort included 22 NICM patients (13 DCM, 9 CS) that underwent pre-procedural CMR prior to ventricular tachycardia (VT) ablation between November 2018 to May 2023. LGE images were delineated into sub-endocardial, mid-myocardial, and sub-epicardial layers, categorized into anterior, lateral, inferior, and septal walls based on the AHA 17 segment model for LV myocardial segmentation. CMR signal intensities were standardized as z-scores: z = (x−μ)/σ, where x is the MRI signal intensity for a specific myocardial segment and layer, and μ and σ are the mean and standard deviation across all myocardial segments and layers of the LV, to characterize regional intensity variations. Results/Data: Compared to patients with DCM, those with CS exhibited significantly higher CMR signal intensity z-scores in the septum (β=0.32, p=0.009), particularly in the endocardial third of the right ventricular side (β=0.56, p=0.001). A z-score greater than 0.40 in this area was associated with a CS diagnosis, with an area under the ROC curve of 0.692 in 5-fold cross-validation. Conclusions: Patients with CS exhibit higher affinity for contrast in the septum, particularly on the RV endocardium. Standardized analysis of CMR signal intensities provides a novel, quantitative method for distinguishing CS from DCM, with the former exhibiting higher CMR signal intensity z-scores in the septum.

Abstract 4147644: Regional Distribution and Overlap of Left Atrial Epicardial Adipose Tissue and Fibrosis

Background: Left atrial epicardial adipose tissue (LA-EAT) and fibrosis are implicated in the pathophysiology of the atrial fibrillation (AF) substrate. Hypothesis: The distribution of fibrosis and LA-EAT differs across various regions of the LA. Aims: Ascertain whether high fibrosis regions coincide with high EAT regions. Identify specific anatomical regions in which LA-EAT and fibrosis tend to co-localize. Methods: LA-EAT was imaged using MRI Dixon sequences and images were segmented axially using the software contouring tools. LGE-MRI was used for LA fibrosis quantification and segmentation. The regions of fibrosis and fat were then co-registered with the atrial geometry and projected onto the left atrial shell. A universal atrial coordinates (UAC) approach was then used to systematically delineate anatomical LA regions in a manner independent of inter-individual variabilities in scale and shape. Specifically, the LA was divided into 10 regions (left and right versions of: posterior-inferior, posterior-superior, roof, anterior-superior, and anterior-inferior). Levene’s test and ANOVA were used to check for any statistically significant variability. Results: Nineteen patients were recruited (94% male; median age 72[62-74]; BMI 32.1±6.3; LA volume index 71.0±21.5mL/m 2 ).LA fibrosis was highest in the left posterior superior segment, right anterior inferior, and left posterior inferior (39.3±16.2%,34.7±26.8%, and 25.7±20.9% respectively;F=1.9,p=0.15). LA-EAT was most prominent in the right anterior superior, left posterior inferior, and left anterior inferior segments (38.1±25.3%,35.6±18.4%,and 33.1±18.9% respectively; F=0.94, p= 0.39). Spatial co-localization of fat and fibrosis was highest in the left posterior inferior region (8.95±2.1%), followed by the left posterior superior, right anterior superior, and left anterior inferior region (7.9±2.1%,7.2±1.7%,and 6.3±1.6% respectively; F= 0.29, p=0.83 ). Conclusion: LA fibrosis and EAT have differing regional preponderances, impacting the AF substrate through infiltrative and paracrine effects. This fibro-fatty remodeling creates structural and conduction abnormalities, forming stable reentrant circuits and perpetuating AF by providing varied pathways and refractory periods.

Relationship between Cardiometabolic index and endometriosis in a US nationally representative sample: results from NHANES 1999-2006

BackgroundEndometriosis is an estrogen-dependent gynecological endocrine condition and a systemic inflammatory disease associated to improper lipid metabolism and increased cardiovascular risk. The Cardiometabolic Index (CMI) is a novel indicator representing visceral adipose tissue distribution and metabolic dysfunction, integrating lipid metabolism indicators and the waist-to-height ratio. While anomalies in lipid metabolism are often associated with (BMI) Body Mass Index, literature consistently shows a negative link between endometriosis and female BMI, and some studies have found that endometriosis is one of the few reproductive diseases not persistently positively correlated with obesity. Given the limitations of BMI, a comprehensive index like CMI is crucial for exploring the incidence of endometriosis. Currently, research on the correlation between CMI and endometriosis is lacking, prompting this study to investigate this association.ObjectiveTo investigate the association between the CMI and the risk of having endometriosis in a sample representing the entire U.S. population.Study designA cross-sectional analysis was conducted using data from four cycles of the National Health and Nutrition Examination Survey (NHANES) spanning the years 1999 to 2006. The study included individuals aged 20 to 54 with a documented history of endometriosis and complete CMI data. Logistic regression analysis, subgroup and interaction analyses, smooth curve fitting, and restricted cubic splines (RCS) were utilized to examine the association between CMI and endometriosis.ResultsThe study found that individuals with higher CMI had an increased probability of developing endometriosis. This relationship remained significant after adjusting for potential confounders such as age, ethnicity, Poverty Income Ratio (PIR), drinking, smoking, education level, and marital status. The fully adjusted model revealed a positive correlation between CMI and endometriosis (OR = 1.21; 95% CI, 1.04–1.40, p < 0.05). Subgroup and interaction analyses showed no significant effect modification by age, BMI, PIR, hypertension, drinking, smoking, or menarche age (all p-values for interaction > 0.05).ConclusionOur study shows a link between CMI and the chance of getting endometriosis.Due to the common occurrence of endometriosis and the lack of clarity surrounding their cause, more study is needed to confirm our results and find out if CMI could be used as a warning sign for endometriosis.

Ocular Tissue Distribution of Omidenepag Isopropyl in Rabbits and Cynomolgus Monkeys.

To evaluate the ocular distribution of omidenepag isopropyl (OMDI) and its active form omidenepag (OMD), an EP2 receptor agonist, after topical administration of OMDI into rabbit and monkey eyes, and to determine whether OMDI and OMD interact with target receptors or enzymes of other antiglaucoma agents. Both eyes of six rabbits and of 14 monkeys were topically instilled with 0.03% [14C]-OMDI. Rabbits were sacrificed after one to four hours, and ocular tissues were collected. Monkeys were sacrificed after 0.25 to 24 hours, and blood and ocular tissues were collected. Radioactivity was measured in each sample. The interactions of OMDI and OMD with the receptors and enzymes associated with the mechanisms of action of other antiglaucoma agents were evaluated. Most radioactivity applied to rabbit eyes was recovered as OMD from the cornea, aqueous humor, and iris-ciliary body. Similarly, high concentrations of radioactivity were observed in monkey cornea, bulbar/palpebral conjunctiva, and trabecular meshwork. OMD bound to EP2 receptors, but neither OMD nor OMDI bound to α2A, β1, and β2 adrenergic receptors or inhibited enzymatic activities of CA1 and CA2. OMD and OMDI had little or no effect on ROCK1 and ROCK2. OMDI rapidly permeates rabbit and monkey corneas and is converted to OMD, which distributes into anterior ocular tissues. Neither OMD nor OMDI interacted with the target receptors or enzymes of other antiglaucoma agents, suggesting that OMD interacts highly selectively with EP2 receptors. OMDI is a specific antiglaucoma agent that interacts selectively with ocular EP2 receptors.

Targeted Therapy of Antibody-Induced Autoimmune Arthritis Using Peptide-Guided Nanoparticles

Rheumatoid arthritis (RA) is an autoimmune disease characterized by chronic inflammation of the joints and it affects over 18 million people worldwide. Despite the availability of a variety of potent drugs for RA, over 30–40 percent of patients fail to achieve adequate remission, and many patients suffer from systemic adverse effects. Thus, there is an urgent need for a joint-targeted drug delivery system. Nanotechnology-based drug delivery methods offer a promising resource that is largely untapped for RA. Using the T cell-driven rat adjuvant-induced arthritis (AA) model of human RA, we developed a peptide-targeted liposomal drug delivery system for arthritis therapy. It was based on a novel joint-homing peptide ART-2 to guide liposomes entrapping dexamethasone (Dex) to arthritic joints of rats, and this approach was more effective in suppressing arthritis than the unpackaged (free) drug. To de-risk the translation of our innovative drug delivery technology to RA patients, we undertook the validation of ART-2-liposomal delivery in a genetically and mechanistically distinct arthritis model in mice, the collagen antibody-induced arthritis (CAIA) model. Using live imaging for tissue distribution of liposomes in vivo, immunohistochemistry of paws for cellular binding of ART-2, and liposomal Dex delivery, our results fully validated the key findings of the rat model, namely, preferential homing of peptide-functionalized liposomes to arthritic joints compared to healthy joints, and higher efficacy of liposomal Dex than free Dex. These results offer a proof-of-concept for the benefits of targeted drug delivery to the joints and its potential translation to RA patients.

Tumor growth and vascular redistribution contributes to the dosimetric preferential effect of microbeam radiotherapy: a Monte Carlo study

The radiobiological mechanisms behind the favorable response of tissues to microbeam radiation therapy (MRT) are not fully described yet. Among other factors, the differential action to tumor and normal tissue vasculature is considered to contribute to MRT efficacy. This computational study evaluates the relevance of tumor growth stage and associated vascular redistribution to this effect. A multiscale approach was employed with two simulation softwares: TOPAS and CompuCell3D. Segmentation images of the angioarchitecture of a non-bearing tumor mouse brain were used. The tumor vasculature at different tumor growth stages was obtained by simulating the tumor proliferation and spatial vascular redistribution. The radiation-induced damage to vascular cells and consequent change in oxygen perfusion were simulated for normal and tumor tissues. The multiscale model showed that oxygen perfusion to tissues and vessels decreased as a function of the tumor proliferation stage, and with the decrease in uniformity of the vasculature spatial distribution in the tumor tissue. This led to an increase in the fraction of hypoxic (up to 60%) and necrotic (10%) tumor cells at advanced tumor stages, whereas normal tissues remained normoxic. These results showed that tumor stage and spatial vascular distribution contribute to the preferential effect of MRT in tumors.

A genome engineered tool set for Drosophila TGF-β/BMP signaling studies.

Ligands of the TGF-β/BMP superfamily are critically involved in the regulation of growth, patterning and organogenesis and can act as long-range morphogens. Essential for understanding TGF-β/BMP signaling dynamics and regulation are tools that allow monitoring and manipulating pathway components expressed at physiological levels and endogenous spatiotemporal patterns. We used genome engineering to generate a comprehensive library of endogenously epitope- or fluorescently-tagged versions of receptors, co-receptors, transcription factors and key feedback regulators of the Drosophila BMP and Activin signaling pathways. We demonstrate that the generated alleles are biologically active and can be utilized for assessing tissue and subcellular distribution of the corresponding proteins. Further, we show that the genomic platforms can be used for in locus structure-function and cis-regulatory analyses. Finally, we present a complementary set of protein binder-based tools, which allow visualization as well as manipulation of the stability and subcellular localization of epitope-tagged proteins, providing new tools for the analysis of BMP signaling and beyond.

Bioaccumulation and tissue distribution of pharmaceuticals and their transformation products in fish along the pollution gradients of a wastewater-impacted river

A field study on the occurrence and distribution of forty-three pharmaceutically active compounds (PhACs) in water and fish samples from anthropogenically impacted section of the Sava River (Croatia) was performed to estimate the importance of bioaccumulation for the environmental risk assessment of PhACs. The study was performed using a highly specific LC-MS/MS method, tailored to include the most prominent PhACs from different therapeutic categories as well as their major metabolites and/or transformation products (TPs). The results revealed a widespread occurrence of PhAC residues both in water and fish samples with a large spatial variability reflecting the distance from the dominant wastewater discharges. The most prominent PhAC categories in less polluted upstream part of the river were common psychostimulants caffeine and cotinine, therapeutic opioids and cardiovascular drugs, while in the river section affected by the local municipal and industrial wastewater inputs, antibiotic drugs became clearly predominant, especially in fish tissue samples. The apparent bioconcentration factors (BCFs) of investigated PhACs varied over several orders of magnitude, from 0.02 ± 0.01 Lkg−1 for O-desmethyl tramadol in fish muscle to 784 ± 260 Lkg−1 for terbinafine in fish liver, indicating rather large differences in their bioconcentration potential and affinity to different tissues, with the tissue-specific BCFs increasing in the following order: muscle < gills < gonads < heart < liver < kidneys. The bioconcentration potential of most of the PhACs included in this study was only low to moderate however moderately high BCFs of certain PhACs (e.g. sertraline, terbinafine, loratadine, diazepam and azithromycin) in some tissues should be taken into consideration when assessing their potential environmental risks. Moreover, it was shown that BCFs could be strongly affected by biotransformation in fish. Risk prioritization based on risk quotient (RQ) and ToxPi index, revealed antibiotics, in particular azithromycin, and therapeutic psychoactive substances as the most hazardous pharmaceutical contaminants in the Sava River.

Depletion of conventional CD4+ T cells is required for robust priming and dissemination of tumor antigen-specific CD8+ T cells in the setting of anti-CD4 therapy

BackgroundOvercoming immune suppression is a major barrier to eliciting potent CD8+ T cell responses against cancer. Treatment with anti-CD4 monoclonal antibody is an effective means for eliminating CD4+Foxp3+ regulatory (Treg)...

Modulating versatile pathways using a cleavable PEG shell and EGFR-targeted nanoparticles to deliver CRISPR-Cas9 and docetaxel for triple-negative breast cancer inhibition.

Human antigen R (HuR), an RNA-binding protein, is implicated in regulating mRNA stability and translation in cancer, especially in triple-negative breast cancer (TNBC), a highly aggressive form. CRISPR/Cas9-mediated HuR knockout (HuR CRISPR) presents a promising genetic therapeutic approach, but it encounters transfection limitations. Docetaxel (DTX), an effective cytotoxic agent against metastatic breast cancer (BC), faces challenges related to vehicle-associated adverse events in DTX formulations. Therefore, we designed multifunctional nanoparticles with pH-sensitive PEG derivatives and targeting peptides to enable efficient HuR CRISPR and DTX delivery to human TNBC MDA-MB-231 cells and tumor-bearing mice. Our findings indicated that these nanoparticles displayed pH-responsive cytotoxicity, precise EGFR targeting, efficient tumor penetration, successful endosomal escape, and accurate nuclear and cytoplasmic localization. They also demonstrated the ability to spare normal cells and prevent hemolysis. Our study concurrently modulated multiple pathways, including EGFR, Wnt/β-catenin, MDR, and EMT, through the regulation of EGFR/PI3K/AKT, HuR/galectin-3/GSK-3β/β-catenin, and P-gp/MRPs/BCRP, as well as YAP1/TGF-β/ZEB1/Slug/MMPs. The combined treatment arrested the cell cycle at the G2 phase and inhibited EMT, effectively impeding tumor progression. Tissue distribution, biochemical assays, and histological staining revealed the enhanced safety profile of pH-responsive PEG- and peptide-modified nanoformulations in TNBC mice. The DTX-embedded and peptide-modified nanoparticles mitigated the side effects of DTX, enhanced cytotoxicity in TNBC MDA-MB-231 cells, and exhibited remarkable antitumor efficacy and safety in TNBC-bearing mice with HuR CRISPR deletion. Collectively, the combination therapy of DTX and CRISPR/Cas9 offers an effective platform for delivering antineoplastic agents and gene-editing systems to combat tumor resistance and progression in TNBC.

Characterization of the C5H11ORF96 gene in chickens: cloning, tissue distribution and investigation of its potential function in stress response regulation

Characterization of the C5H11ORF96 gene in chickens: cloning, tissue distribution and investigation of its potential function in stress response regulation

A comparative study for organophosphate triesters and diesters in mice via oral gavage exposure: Tissue distribution, excreta elimination, metabolites and toxicity

Organophosphate triesters (tri-OPEs) and diesters (di-OPEs) may threaten human health through dietary intake, whereas little information is available about their fate in mammals. Herein, mice exposure experiments were carried out through gavage with six tri-OPEs and six di-OPEs, respectively. The residual levels of di-OPEs in mice were generally higher than those of tri-OPEs. The residual di-OPEs mainly distributed in the liver and blood while the most tri-OPEs remained in stomach, indicating easier transfer and lower metabolism levels of di-OPEs. The accumulation of tri- and di-OPEs with large octanol–water partition coefficients and long carbon chain were observed in tissues and feces, implying that the elimination of these OPEs through fecal excretion is an important elimination pathway. A total of 86 OPE metabolites were found in murine urine and feces, 57 of which were identified for the first time. For tri-OPEs, carboxylated OPEs had higher peak intensities and fewer interference factors among the metabolites, which could serve as ideal biomarkers. The predicted oral median lethal doses of OPEs and corresponding metabolites showed an increased toxicity of some hydroxylated OPEs and di-OPEs, needing further attention. These results provided new insights and evidence on the fates and biomarkers of OPEs exposure for mammals.

Comprehensive mapping of somatotroph pituitary neuroendocrine tumour heterogeneity using spatial and single-cell transcriptomics.

Pituitary neuroendocrine tumours (PitNETs) are common intracranial tumours that are highly heterogeneous with unpredictable growth patterns. The driver genes and mechanisms that are crucial for tumour progression in somatotroph PitNETs are poorly understood. In this study, we performed integrative spatial transcriptomics (ST) and single-cell RNA sequencing (scRNA-seq) analysis on somatotroph tumours and normal pituitary samples to comprehensively characterize the differences in cellular characteristics. By analyzing combined copy number variations (CNVs), tumour tissues were divided into two regions, which included the CNVhigh and CNVlow areas. The protumour genes DLK1 and RCN1 were highly expressed in the CNVhigh area, which might be related to tumour progression and could be targeted for precision therapy. We also found that the transforming growth factor beta signalling pathway participated in tumour progression and identified heterogeneity in the expression profiles of key genes. We assessed the intertumoral and intratumoral heterogeneity in somatotroph PitNETs and emphasized the importance of individualized treatment. In summary, we visualized the cellular distribution and transcriptional differences in normal pituitary and somatotroph PitNETs by ST and scRNA-seq for the first time. This study provides a strong theoretical foundation to comprehensively understand the crucial mechanisms involved in tumour progression and develop new strategies to treat somatotroph PitNETs. The first-ever visualization of cellular distributions in normal and tumor pituitary tissues. The inter- and intra-tumoral transcriptomic heterogeneity of somatotroph PitNETs was comprehensively revealed. Identification of potential protumor factors and critical signaling pathways, opening new avenues for therapeutic intervention.

Interactions between glucagon like peptide 1 (GLP-1) and estrogens regulates lipid metabolism

Obesity, characterized by excessive fat accumulation in white adipose tissue (WAT), is linked to numerous health issues, including insulin resistance (IR), and type 2 diabetes mellitus (DM2). The distribution of adipose tissue differs by sex, with men typically exhibiting android adiposity and pre-menopausal women displaying gynecoid adiposity. After menopause, women have an increased risk of developing android-type obesity, IR, and DM2. Glucagon-like peptide 1 (GLP-1) receptor agonists (GLP-1RAs) are important in treating obesity and DM2 by regulating insulin secretion, impacting glucose and lipid metabolism. GLP-1Rs are found in various tissues including the pancreas, brain, and adipose tissue. Studies suggest GLP-1RAs and estrogen replacement therapies have similar effects on tissues like the liver, central nervous system, and WAT, probably by converging pathways involving protein kinasesTo investigate these interactions, female rats underwent ovariectomy (OVR) to promote a state of estrogen deficiency. After 20 days, the rats were euthanized and the tissues were incubated with 10 μM of liraglutide, a GLP-1RA. Results showed significant changes in metabolic parameters: OVR increased lipid catabolism in perirenal WAT and basal lipolysis in subcutaneous WAT, while liraglutide treatment enhanced stimulated lipolysis in subcutaneous WAT. Liver responses included increased stimulated lipolysis with liraglutide. Transcriptome analysis revealed distinct gene expression patterns in WAT of OVR rats and those treated with GLP-1RA, highlighting pathways related to lipid and glucose metabolism. Functional enrichment analysis showed estrogen’s pivotal role in these pathways, influencing genes involved in lipid metabolism regulation.Overall, the study underscores GLP-1RA acting directly on adipose tissues and highlights the complex interactions between GLP-1 and estrogen in regulating metabolism, suggesting potential synergistic therapeutic effects in treating metabolic disorders like obesity and DM2.

LITERATURE REVIEW ON ANALYTICAL METHODS FOR QUANTIFICATION OF SUVOREXANT

This review summarizes various analytical methods reported for the quantification of suvorexant, a novel dual orexin receptor antagonist used for the treatment of insomnia. The methods discussed include chromatographic techniques such as HPLC-MS/MS, GC-MS, and HPTLC. The review highlights the sensitivity, specificity, and linearity range of these methods. Additionally, the review explores the formulation and tissue distribution of suvorexant.

Synthesis, biological evaluation and mechanism study based on network pharmacology of amino acids esters of 20(S)-protopanaxadiol as novel anticancer agents

As one of the metabolites of ginseng, 20(S)-protopanaxadiol (PPD) is a compound with dammarane-type tetracyclic triterpene, which performs a wide range of anticancer activities. In this study, PPD was used as a lead. A series of compounds were synthesized respectively with 11 amino acids through esterification and were evaluated for their cytotoxicity against several cancer cell lines. One of the synthetic products (PL) exhibited potent inhibitory effect on Huh-7 cells relative to that of PPD in vitro. Subsequently, the Annexin V-FITC /PI staining assay was used to verify that PL induced apoptosis of Huh-7 cells in a dose-dependent manner. A UPLC-Q/TOF-MS analysis method was established and validated for assessing pharmacokinetic properties after the administration of PPD and PL in rats. The results showed that compared with PPD, T1/2of PL in rats was prolonged, and the peak time was delayed, resulting in broader tissue distribution of the compound in the body. In addition, the targets of PL against several cancers were predicted and analyzed via network pharmacology. Molecular docking simulations demonstrated that PL interacted with the active sites of the above targets. In conclusion, this study provided a theoretical basis for the development and clinical application of anti-tumor activity of PPD.