Target Tissues Research Articles

Current Updates of CRISPR/Cas System and Anti-CRISPR Proteins: Innovative Applications to Improve the Genome Editing Strategies.

The Clustered Regularly Interspaced Short Palindromic Repeats (CRISPR)/CRISPR-associated sequence (CRISPR/Cas) system is a cutting-edge genome-editing tool employed to explore the functions of normal and disease-related genes. The CRISPR/Cas system has a remarkable diversity in the composition and architecture of genomic loci and Cas protein sequences. Owing to its excellent efficiency and specificity, this system adds an outstanding dimension to biomedical research on genetic manipulation of eukaryotic cells. However, safe, efficient, and specific delivery of this system to target cells and tissues and their off-target effects are considered critical bottlenecks for the therapeutic applications. Recently discovered anti-CRISPR proteins (Acr) play a significant role in limiting the effects of this system. Acrs are relatively small proteins that are highly specific to CRISPR variants and exhibit remarkable structural diversity. The in silico approaches, crystallography, and cryo-electron microscopy play significant roles in elucidating the mechanisms of action of Acrs. Acrs block the CRISPR/Cas system mainly by employing four mechanisms: CRISPR/Cas complex assembly interruption, target-binding interference, target cleavage prevention, and degradation of cyclic oligonucleotide signaling molecules. Engineered CRISPR/Cas systems are frequently used in gene therapy, diagnostics, and functional genomics. Understanding the molecular mechanisms underlying Acr action may help in the safe and effective use of CRISPR/Cas tools for genetic modification, particularly in the context of medicine. Thus, attempts to regulate prokaryotic CRISPR/Cas surveillance complexes will advance the development of antimicrobial drugs and treatment of human diseases. In this review, recent updates on CRISPR/Cas systems, especially CRISPR/Cas9 and Acrs, and their novel mechanistic insights are elaborated. In addition, the role of Acrs in the novel applications of CRISPP/Cas biotechnology for precise genome editing and other applications is discussed.

Clinical Pharmacology Of Selective Oestrogen Receptor Modulators In The Prevention Of Breast Cancer, Osteoporosis, Coronary Heart Disease, Alzheimer's Disease, Use In Hormone Replacement Therapy, Cholesterol Metabolism

Oestrogen is the key regulatory hormone for the development and maintenance of reproductive functions and affects various physiological systems, including the skeletal, cardiovascular, and central nervous systems. oestrogen replacement therapy (ERT) is highly effective in postmenopausal women, addressing menopausal symptoms and providing beneficial effects on osteoporosis and coronary heart disease. However, ERT is associated with significant side effects, such as an increased risk of breast cancer. To mitigate these risks, selective oestrogen receptor modulators (SERMs) have been developed. SERMs are a growing class of synthetic nonsteroidal compounds that act as oestrogen receptor agonists or antagonists in a tissue-specific manner. They can selectively activate or inhibit oestrogen receptors in different tissues throughout the body. SERMs are used to treat a variety of conditions, including ER-positive breast cancer, osteoporosis, coronary heart disease, Alzheimer’s disease, and cholesterol metabolism disorders in postmenopausal women. Specific SERMs are employed for different conditions: raloxifene for osteoporosis, tamoxifen and toremifene for breast cancer, clomiphene for the induction of ovulation, and ormeloxifene for oral contraception. Arzoxifene is used for altering cholesterol metabolism. Each SERM forms a unique complex with oestrogen receptors, influencing target tissues and resulting in a tissue-selective pharmacological profile. This unique mechanism of action translates into specific safety and efficacy profiles for each SERM. The surprising pharmacology of SERMs has sparked interest in developing new selective medications, allowing for precise treatment of various diseases. This article reviews the pharmacological properties of SERMs and explains how they differ from oestrogen in their actions.

Nanoparticle-based Gene Therapy for Neurodegenerative Disorders.

Neurological disorders present a formidable challenge in modern medicine due to the intricate obstacles set for the brain and the multipart nature of genetic interventions. This review article delves into the promising realm of nanoparticle-based gene therapy as an innovative approach to addressing the intricacies of neurological disorders. Nanoparticles (NPs) provide a multipurpose podium for the conveyance of therapeutic genes, offering unique properties such as precise targeting, enhanced stability, and the potential to bypass blood-brain barrier (BBB) restrictions. This comprehensive exploration reviews the current state of nanoparticle-mediated gene therapy in neurological disorders, highlighting recent advancements and breakthroughs. The discussion encompasses the synthesis of nanoparticles from various materials and their conjugation to therapeutic genes, emphasizing the flexibility in design that contributes to specific tissue targeting. The abstract also addresses the low immunogenicity of these nanoparticles and their stability in circulation, critical factors for successful gene delivery. While the potential of NP-based gene therapy for neurological disorders is vast, challenges and gaps in knowledge persist. The lack of extensive clinical trials leaves questions about safety and potential side effects unanswered. Therefore, this abstract emphasizes the need for further research to validate the therapeutic applications of NP-mediated gene therapy and to address nanosafety concerns. In conclusion, nanoparticle-based gene therapy emerges as a promising avenue in the pursuit of effective treatments for neurological disorders. This abstract advocates for continued research efforts to bridge existing knowledge gaps, unlocking the full potential of this innovative approach and paving the way for transformative solutions in the realm of neurological health.

Anticancer behavior of cyclometallated iridium(III)-tributyltin(IV) carboxylate schiff base complexes with aggregation-induced emission

Cyclometallated iridium(III) and organotin(IV) carboxylate complexes have shown potential application value in the field of anticancer. However, the widespread aggregation-caused quenching (ACQ) effect of these complexes is not conducive to the exploration of their targeting and anticancer mechanism, and the idea of aggregation-induced emission (AIE) effect can effectively solve this problem. Then, AIE-activated cyclometallated iridium(III)-tributyltin(IV) carboxylate Schiff base complexes were designed and prepared in this study. Complexes exhibited AIE effect in highly concentrated solution or aggregative state, which facilitated the investigation of subcellular tissue targeting (mitochondria) and cell morphology. Compared with cyclometallated iridium(III) complex and tributyltin(IV) carboxylate monomers, these complexes showed the better in-vitro anti-proliferative activity toward A549 cells, confirming the favorable synergistic anticancer activity. Even for A549/DDP (cisplatin-resistance) cells, these complexes also exhibited the better activity. In addition, complexes showed a mitochondrial apoptotic pathway. Therefore, cyclometallated iridium(III)-tributyltin(IV) carboxylate Schiff base complexes can be used as the potential substitutes for platinum-based drugs and gain further application.

Intercellular Adhesion Disorders In Tumorigenesis

The review discusses the problem of adhesion impairment in the course of tumorigenesis and aging. We hypothesize that impairment of homophilic intercellular adhesion in the target tissue results in developing conditions, which are favorable for malignancy, invasion, and metastases. Like a phoenix vanishing during the initiation of a primary tumor by breaking contacts between identical cells, adhesion molecules reappear with a new quality (the phoenix rising mechanism), thereby causing invasive and metastatic behavior of tumor cells. Due to this, primary tumor cells acquire motility and the ability to form metastases, which are the cause of most cancer deaths. At the same time, the provision of adhesive bonds between cancer cells and immune effector cells can also be controlled by one of the main neurotransmitters, dopamine (DA). The discovery of peripheral DA in lymphocytes gave grounds to the assumption that DA is involved in the infiltration of tumor leukocytes. DA receptors are found on cells of the adaptive (specific) immune response: T and B lymphocytes. Direct communication between brain DA and peripheral DA is crucial in modulating immune function. Peripheral DA mediates differentiation, binding to tumor cells, and cytotoxicity of CD8+ T cells. The review also confirms the need for the development of adhesion pharmaceutical agents. The disruption of intercellular adhesion in the target tissue and the general deficiency of immune surveillance can be controlled by central mechanisms involving brain DA, which is capable of regulating the active phase of immune responses against the tumor by means of adhesive interactions in the immune system, interfering with the process and thereby interrupting the development of a malignant neoplasm initiated by a local mutation in the tissue. The concept reveals the stress mechanism of cancer etiology and creates prospects for new methods of diagnostics, prevention and treatment of tumors, which can become another step towards solving the problem of malignant neoplasms.

Duck circovirus regulates the expression of duck CLDN2 protein by activating the MAPK-ERK pathway to affect its adhesion and infection.

Although duck circovirus (DuCV) poses a widespread infection and a serious hazard to the duck industry, the molecular mechanisms underlying DuCV infection and transmission remain elusive. We initially demonstrated vertical transmission of DuCV through female breeding ducks by simulating natural infection. Furthermore, a differentially expressed membrane protein CLDN2 was identified on the DuCV-infected oviduct of female ducks, and its extracellular loop structural domains EL1 and EL2 were identified as the interaction sites of DuCV Cap proteins. Moreover, the binding of DuCV Cap to CLDN2 triggered the intracellular MAPK-ERK pathway and activated the downstream transcription factor SP5. Importantly, we demonstrated that intracellular Cap also interacts with SP5, leading to upregulation of CLDN2 transcription and facilitating enhanced adherence of DuCV to target tissue, thereby promoting viral infection and transmission. Our study sheds light on the molecular mechanisms underlying vertical transmission of DuCV, highlighting CLDN2 as a promising target for drug development against DuCV infection.

Evaluation of indirect indices in the insulin resistance assessment in patients with different body mass index

Introduction: Insulin resistance (IR) is a complex pathophysiological condition with multifactorial etiology characterized by a reduced responsiveness of target tissues to insulin (INS). Indirect indices based on mathematical models and derived from laboratory parameters have become increasingly popular in the past two decades. In this study, we evaluated their ability to predict IR in a population with different body mass index (BMI). Methods: The matched case–control study was conducted in 2021 and 2022. Secondary data from 129 subjects were obtained from medical records, including demographic characteristics, anthropometric measurements, and biochemical laboratory test results. The studied group consisted of 91 subjects with a suspected diagnosis of IR who were further categorized according to BMI, while control group consisted from 38 age- and gender-matched subjects. Six widely used indirect indices were calculated: Homeostatic Model Assessment for IR (HOMA-IR), quantitative insulin sensitivity check index (QUICKI), McAuley index (MCAi), metabolic score for IR (METS-IR), triglyceride to glucose index (TyG), and TyG to BMI (TyG-BMI). Results: Significant differences between the subgroups were found in the mean values for HOMA-IR, TyG, TyG-BMI, and METS- IR, while the control group had the highest mean values for the indirect indices QUICKI and MCAi (p < 0.001). HOMA-IR, TyG, and TyG-BMI showed statistical significance in predicting IR regardless of BMI (p < 0.05). In the obese group, TyG-BMI had good predictive power for discriminating IR (area under the curve [AUC] = 0.820), with a sensitivity and specificity of 84.1% and 87.7%, respectively. HOMA-IR showed moderate predictive power to discriminate IR in the obese group (AUC = 0.720), with a sensitivity and specificity of 70.4% and 89.1%, respectively. Conclusion: As IR is a multifactorial disease, indirect indices combining laboratory and anthropometric data can significantly help in predicting and mitigating complications.

Cone Beam Computed Tomography-Guided Navigation Bronchoscopy with Augmented Fluoroscopy for the Diagnosis of Peripheral Pulmonary Nodules: A Step-by-Step Guide.

Cone beam computed tomography-guided navigation bronchoscopy (CBCT-NB) with augmented fluoroscopy (AF) guidance represents a minimally invasive endobronchial technique for diagnosing small, peripheral pulmonary lesions. This approach is characterized by its high diagnostic accuracy and low complication risk. Current pilot trials are exploring the application of localized therapies using this innovative approach. This report aims to provide a detailed procedural guide for performing CBCT-NB with AF guidance as the only tool for navigation and image guided biopsy. We outline the procedural steps involved in the CBCT-NB procedure for diagnosing peripheral pulmonary lesions, supported by specific intra-procedural clinical video footage. The steps include (1) preprocedural considerations, (2) a detailed procedural workflow encompassing navigation to the target lesion, (3) position confirmation and tissue acquisition, and (4) postprocedural follow-up. CBCT-NB with AF guidance is a safe and precise stand-alone navigation modality that offers high-resolution real-time 3D imaging, enhancing the diagnosis and potential treatment of peripheral pulmonary nodules.

ROS-responsive nanomicelles encapsulating celastrol ameliorate pressure overload-induced cardiac hypertrophy by regulating the NF-κB signaling pathway

Celastrol (CEL) belongs to the group of non-steroidal immunosuppressants with the potential to improve cardiac hypertrophy (CH). However, the poor biocompatibility and low bioavailability of CEL limit its in vivo application. This study was aimed to develop a targeted drug delivery system that can efficiently and safely deliver CEL to target tissues, providing a research basis for the application of CEL in CH therapy. A novel ROS-sensitive drug-loaded nanomicelle, dodecanoic acid (DA)-phenylboronic acid pinacol ester-dextran polymer encapsulating CEL (DBD@CEL), was synthesized using chemical synthesis. Then, the morphology, particle size, drug-loaded content, and ROS-responsive release behavior of DBD@CEL were studied. Pharmacokinetics and biocompatibility were evaluated using healthy mice. Finally, the ability and mechanism of DBD@CEL in improving CH in vivo were investigated using a mouse CH model. DBD@CEL was successfully prepared with a drug loading of 18.9%. It exhibited excellent stability with an average particle size of 110.0 ± 1.7 nm. Within 48 h, DBD@CEL released only 19.4% in the absence of H2O2, while in the presence of 1 mM H2O2, the release rate increased to 71.5%. Biocompatibility studies indicated that DBD@CEL did not cause blood cell hemolysis, had no impact on normal organs, and did not result in abnormal blood biochemical indicators, demonstrating excellent biocompatibility. In vivo studies revealed that DBD@CEL regulated the activation of NF-κB signaling, inhibits pyroptosis and oxidative stress, and thereby ameliorates CH. The ROS-responsive DBD@CEL nanodrug delivery system enhances the therapeutic activity of CEL for CH, providing a promising drug delivery system for the clinical treatment of CH.

The impact of host microRNAs on the development of conserved mutations of SARS-CoV-2.

SARS-CoV-2, the virus responsible for the COVID-19 pandemic, has undergone various genetic alterations due to evolutionary pressures exerted by host cells, including intracellular antiviral mechanisms such as targeting by human microRNAs (miRNAs). This study investigates the impact of miRNAs hsa-miR-3132 and hsa-miR-4650 on the viral genome. Sequence alignment revealed conserved mutations in the binding sites of these miRNAs in adapted strains compared to the original Wuhan-Hu-1 strain, leading to their deletion. Despite modest expression of these miRNAs in SARS-CoV-2 target tissues, their efficacy against mutant strains is reduced due to the loss of binding sites. Structural analysis indicates that the mutant genome is more stable than the Wuhan-Hu-1 genome. Luciferase and virus titration assays demonstrate that hsa-miR-3132 and hsa-miR-4650 effectively target the Nsp3 gene in the Wuhan-Hu-1 strain but not in mutant strains lacking their binding sites. These findings suggest that the observed mutations help the virus evade selective pressure from human miRNAs, contributing to its adaptation.

Nanomedicine in Bladder Cancer Therapy.

Bladder cancer (BC) is one of the most common malignant neoplasms of the genitourinary system. Traditional BC therapies include chemotherapy, targeted therapy, and immunotherapy. However, limitations such as lack of specificity, cytotoxicity, and multidrug resistance pose serious challenges to the benefits of BC therapies. Consequently, current studies focus on the search for new therapeutic solutions. In recent years, there has been a growing interest in using nanotechnology in the treatment of both non-invasive (NMIBC) and invasive bladder cancer (MIBC). Nanotechnology is based on the use of both organic molecules (chitosan, liposomes) and inorganic molecules (superparamagnetic iron oxide nanoparticles) as carriers of active substances. The main aim of such molecules is the targeted transport and prolonged retention of the drug in the target tissue, which increases the therapeutic efficacy of the active substance. This review discusses the numerous types of nanoparticles (including chitosan, polymeric nanoparticles, liposomes, and protein nanoparticles), targeting mechanisms, and approved nanotherapeutics with oncological implications in cancer treatment. We also present nanoformulation applications in phototherapy, gene therapy, and immunotherapy. Moreover, we summarise the current perspectives, advantages, and challenges in clinical translation.

Characterizing Temperature-Dependent Acoustic and Thermal Tissue Properties for High-Intensity Focused Ultrasound Computational Modeling

High-intensity focused ultrasound (HIFU) thermal therapies utilize concentrated sound waves to ablate diseased tissue at precise locations within the body. Computational simulations of HIFU can assist clinicians by predicting the death of target tissues, identifying sensitive healthy tissues that risk thermal damage, and optimizing acoustic power delivery to minimize treatment times and maximize treatment efficacy. Accurate simulations require accurate inputs, and many computational solvers neglect property changes induced by tissue heating during treatment. Additionally, temperature-dependent tissue property data in the literature are relatively scarce. This study presents methodology for characterizing temperature-dependent acoustic and thermal properties in ex vivo porcine muscle tissue. From 20 – 50 °C, speed of sound is found to increase from approximately 1580 – 1620 m/s. The acoustic attenuation coefficient increases for 20 – 50 °C from 0.09 – 0.24 Np/cm at 0.5 MHz and 0.16 – 0.37 Np/cm at 1.6 MHz. Thermal conductivity and thermal diffusivity increase from 0.52 – 0.55 W/m °C and 0.147 – 0.158 mm2/s, respectively, over 20 – 60 °C. Specific heat capacity increases from approximately 3500 – 3800 J/kg °C, over 20 – 80 °C. Each property is consistent with data found in the literature, extends the literature to a larger temperature range, and, for acoustic properties, extends to unique frequencies. Temperature-dependent predictive models are also developed for each of the five properties. This study’s property measurement methodologies can be used to characterize other biological tissues, and the predictive models developed herein will facilitate future efforts in temperature-dependent modeling and uncertainty quantification of HIFU thermal therapies.

Radiofrequency Improves Facial Fine Lines by Thermal Effect: Damage or Just Stimulation?

Radiofrequency (RF) has been widely used for rejuvenation treatments, but without knowledge of the effective temperature of the target tissues or guidance on treatment parameters, it often leads to adverse reactions and pain. The aim of this study was to demonstrate that thermal stimulation can produce facial rejuvenation effects and to determine the optimal bipolar RF treatment parameters for treating facial fine lines. A bipolar RF device combined with CORE Technology was used in this study. Exvivo studies were conducted on both miniature swine and human skin, utilizing thermographic thermometry and histological analysis. Invivo swine studies were conducted using histological analysis and electron microscopy. A clinical trial was conducted, and the results were evaluated using the Alexiades Comprehensive Grading Scale (ACGS) and Global Aesthetic Improvement Scale (GAIS) scales. The bipolar RF technology can produce a significant effect by thermally stimulating collagen within just 2 weeks without causing thermal damage. A clinical trial involving 46 patients showed a noticeable rejuvenating effect of the bipolar RF device, especially on fine wrinkles. This study confirmed that thermal stimulation, rather than thermal damage, is sufficient to achieve rejuvenation effects. The study also found a range of bipolar RF treatment parameters that are both safe and effective for facial fine lines.

A Review on Proniosomes as a Novel Drug Delivery System.

Proniosomes are desiccated versions of surfactant-coated, water-soluble carrier particles. Before usage, on agitation in heated aqueous solution, they are rehydrated in a matter of minutes to create niosomal dispersion. In storage and transit, proniosomes maintain their physical stability. The drug contained within the proniosomes’ vesicular structure increases the drug’s potency, improves its absorption into the target tissue, and lowers its toxicity. Niosomes are technically more promising than liposomes as drug carriers since they are more chemically stable and don’t have many of the glitches of liposomes, like high cost and problems with phospholipid purity. Focusing on proniosome preparation, characterization, and application in targeted drug delivery, this study highlights their general approaches.

Recent Advancements in Drug Delivery System in Lung Cancer

Eliminating the fact that the use of free pharmaceuticals in conventional dosage forms usually involves difficulties in hitting the target site at the appropriate dose after or during a correct time period, medication targeting specific organs and tissues became one of the century's most important initiatives. Thus, finding new drug delivery strategies and modes of action are lines of battle. Difficulties are still encountered with radiation therapy to accurately track the movements of lung tumors while treatments are being admitted. The development of MRI-linac hybrid system prototypes offers the possibility of real-time, ionization-free tumor imaging. This study assesses how well lung tumor tracking algorithms function on five healthy participants' cine-MRI sagittal images. The targets were vascular structures that could be seen. Lung cancer is the leading cause of mortality in the world. This is one of the heterogeneous diseases that include, at the basic level, two main subtypes: small cell lung cancer and non-small cell lung cancer. Less than 20% of patients with LC survive for 5 years on average despite recent improvements in treatment. The effectiveness of existing therapeutic techniques is impaired because of severe off-target effects and innate or acquired medication resistance. The two major types of lung cancers are SCLC and NSCLC; among these, the former is the most frequently diagnosed. Lung cancers account for the highest mortality rate due to cancer. The application of nanomedicines can partially overcome the failures that are associated with the anticancer therapies against NSCLC. One of the nanoparticle subfields that hold out much promise for better delivery while ensuring stability and sufficient bioavailability of administered anticancer medications is nanomedicine.

Packaging of alphavirus-based self-amplifying mRNA yields replication-competent virus through a mechanism of aberrant homologous RNA recombination.

Messenger (m)RNA has taken center stage in vaccine development, gene therapy, and cancer immunotherapy. A next-generation of mRNA is the self-amplifying (sa)mRNA, which induces broad and long-lasting immunity at a lower dose which provides better clinical outcomes in conjunction with fewer adverse effects. SamRNA, also known as "replicon" RNA, encodes the replication machinery of an alphavirus together with an antigen. Efficient delivery of replicon RNA to target tissues can be accomplished by packaging the replicon RNA in virus-like replicon particles (VRPs) via co-transfection of producer cells with defective helper RNA(s) encoding the alphavirus structural proteins. During the manufacture of VRPs, however, there is a potential risk of RNA recombination, which may lead to the formation of replication-competent virus (RCV). To investigate the factors influencing the unwanted RCV formation, we evaluated how sequence homology orchestrates alphavirus RNA recombination. Several combinations of complementing alphavirus replicon and helper RNAs varying in length of sequences overlap were co-transfected in mammalian cells. The culture fluid was serially passaged to detect RCV. Nanopore sequencing of cells after the first passage in combination with amplicon-based Sanger sequencing of RCV in the culture fluid after four passages led to the detection of RNA recombination. RCV was generated between replicon and helper RNAs with sequence homology in either the non-structural or structural genes, whereas RNAs without overlapping gene regions did not generate RCV. Remarkably, no sequence overlap was detected at the recombination junction sites in the RCV genome, suggesting a mechanism of "aberrant homologous RNA recombination." Accordingly, we conclude that the alphavirus RNA recombination process leading to the formation of RCV is homology-assisted and can be prevented by avoiding sequence homology between replicon and helper RNAs.IMPORTANCEThere is a growing interest in the use of self-amplifying (sa)mRNA vectors for next-generation vaccine development, gene therapy, and cancer immunotherapy. The delivery of samRNA in the form of virus-like replicon particles (VRPs) enables efficient delivery of samRNA to target tissue. The production of these VRPs, however, suffers from contamination with replication-competent virus (RCV) that is thought to arise from recombination events between samRNA and helper RNAs for VRP packaging. The presence of RCV in samRNA in the clinical product is undesirable as alphaviruses may cause serious disease in humans. However, the underlying recombination mechanism leading to RCV is currently unknown. In our work, we demonstrate a detailed evaluation of the recombination sites, which indicates that RCV is formed through an unusual mechanism of "aberrant homologous RNA recombination." The results are useful for researchers in the field of RNA vaccine manufacture and delivery.

Minor changes to circulating steroid hormones in female rats after perinatal exposure to diethylstilbestrol or ketoconazole

Current chemical test strategies lack sensitive markers for detecting female reproductive toxicity caused by endocrine disrupting chemicals (EDCs). In search of a potentially sensitive readout, the steroidogenic disrupting effects of the well-known EDCs ketoconazole (KTZ) and diethylstilbestrol (DES) were investigated in vitro and on circulating steroid hormones in perinatally exposed female Sprague-Dawley rats. Twenty-one steroid hormones were analysed using LC-MS/MS in plasma from female rat offspring at postnatal day (PD) 6, 14, 22, 42 and 90. Most circulating steroid hormone levels increased with age except for estrone (E1), estradiol (E2) and backdoor pathway androsterone (ANDROST), which decreased after PD 22. Perinatal exposure to DES did not affect circulating steroid hormone levels at any dose or age compared to controls. KTZ exposure resulted in dose-dependent increase of corticosterone (CORTICO) at PD 6 and PD 14, with statistical significance only at PD 14. In the in vitro gold standard H295R steroidogenesis assay, twenty-one steroid hormones were measured instead of only T and E2. DES had subtle effects on steroidogenesis, whereas KTZ decreased most steroid hormones, but increased CORTICO, progesterone (P4), estriol (E3) initially (around 0.1–1 µM) before decreasing. Our data suggests that circulating steroidomic profiling may not be a sensitive readout for EDC-induced female reproductive toxicity. Further studies are needed to associate H295R assay steroidomic profiles with in vivo profiles, especially in target tissues such as adrenals or gonads. Expanding the H295R steroidogenic assay to include a comprehensive steroidomic profile may enhance its regulatory applicability.

Micelle-encapsulated IR783 for enhanced photothermal therapy in mouse breast cancer

BackgroundPhotothermal therapy, an emerging cancer treatment, selectively eliminates lesions using photothermal compounds that convert light into heat. IR783, a near-infrared fluorescent heptamethine cyanine dye, has been used to achieve selective hyperthermic effects in target tissues via near-infrared irradiation. To implement IR783 as a photothermal agent, IR783 biodistribution must be calibrated to achieve a constant and uniform concentration in target cells. Accordingly, we developed micelle-encapsulated IR783 (IR783 micelles) and evaluated their effectiveness as photothermal drugs. MethodsIn vitro, the photothermic effects of free IR783 and IR783 micelle solutions induced by near-infrared light irradiation were analyzed. Additionally, we investigated the mechanism of cell death mediated by photothermal therapy using free IR783 and IR783 micelles in mouse breast cancer (EMT6) cells. In vivo, the efficacy of photothermal therapy with both free IR783 and IR783 micelles was examined in EMT6-bearing mice. ResultsIn vitro, the temperature of free and micelle-encapsulated IR783 solutions increased after near-infrared irradiation. Near-infrared irradiation with free IR783 and IR783 micelles induced cytotoxicity in cancer cells by generating heat. In vivo, IR783 micelles elicited more preferential tumor tissue uptake and enhanced the antitumor effects of photothermal therapy at a lower light dose relative to free IR783. ConclusionsOverall, these results suggest that IR783 micelles could accumulate in mouse breast cancer tissues and exhibit enhanced antitumor effects when used as a photothermal therapy, with superior effects obtained at 2.1 W/cm2 (252 J/cm2) compared with that of free IR783.

Maternal diet during pregnancy and adaptive changes in the maternal and fetal pancreas have implications for future metabolic health.

Fetal and neonatal development is a critical period for the establishment of the future metabolic health and disease risk of an individual. Both maternal undernutrition and overnutrition can result in abnormal fetal organ development resulting in inappropriate birth size, child and adult obesity, and increased risk of Type 2 diabetes and cardiovascular diseases. Inappropriate adaptive changes to the maternal pancreas, placental function, and the development of the fetal pancreas in response to nutritional stress during pregnancy are major contributors to a risk trajectory in the offspring. This interconnected maternal-placental-fetal metabolic axis is driven by endocrine signals in response to the availability of nutritional metabolites and can result in cellular stress and premature aging in fetal tissues and the inappropriate expression of key genes involved in metabolic control as a result of long-lasting epigenetic changes. Such changes result is insufficient pancreatic beta-cell mass and function, reduced insulin sensitivity in target tissues such as liver and white adipose and altered development of hypothalamic satiety centres and in basal glucocorticoid levels. Whilst interventions in the obese mother such as dieting and increased exercise, or treatment with insulin or metformin in mothers who develop gestational diabetes, can improve metabolic control and reduce the risk of a large-for-gestational age infant, their effectiveness in changing the adverse metabolic trajectory in the child is as yet unclear.

A DNA-free and genotype-independent CRISPR/Cas9 system in soybean

Abstract Here, we report a smart genome editing system for soybean (Glycine max) using the in planta bombardment-ribonucleoprotein (iPB-RNP) method without introducing foreign DNA or requiring traditional tissue culture processes such as embryogenesis and organogenesis. Shoot apical meristem (SAM) of embryonic axes was used as the target tissue for genome editing because the SAM in soybean mature seeds has stem cells and specific cell layers that develop germ cells during the reproductive growth stage. In the iPB-RNP method, the RNP complex of the CRISPR/Cas9 system was directly delivered into SAM stem cells via particle bombardment, and genome-edited plants were generated from these SAMs. Soybean allergenic gene Gly m Bd 30K was targeted in this study. Many E0 (the first generation of genome-edited) plants in this experiment harbored mutant alleles at the targeted locus. Editing frequency of inducing mutations transmissible to the E1 generation was approximately 0.4 to 4.6 % of all E0 plants utilized in various soybean varieties. Furthermore, simultaneous mutagenesis by iPB-RNP method was also successfully performed at other loci. Our results offer a practical approach for both plant regeneration and DNA-free genome editing achieved by delivering RNP into the SAM of dicotyledonous plants.