GeneAgent: Self-verification Language Agent for Gene Set Knowledge Discovery using Domain Databases.

Gene set knowledge discovery is essential for advancing human functional genomics. Recent studies have shown promising performance by harnessing the power of Large Language Models (LLMs) on this task. Nonetheless, their results are subject to several limitations common in LLMs such as hallucinations. In response, we present GeneAgent, a first-of-its-kind language agent featuring self-verification capability. It autonomously interacts with various biological databases and leverages relevant domain knowledge to improve accuracy and reduce hallucination occurrences. Benchmarking on 1,106 gene sets from different sources, GeneAgent consistently outperforms standard GPT-4 by a significant margin. Moreover, a detailed manual review confirms the effectiveness of the self-verification module in minimizing hallucinations and generating more reliable analytical narratives. To demonstrate its practical utility, we apply GeneAgent to seven novel gene sets derived from mouse B2905 melanoma cell lines, with expert evaluations showing that GeneAgent offers novel insights into gene functions and subsequently expedites knowledge discovery.

Detection of abdominopelvic lymph nodes in multi-parametric MRI.

Reliable localization of lymph nodes (LNs) in multi-parametric MRI (mpMRI) studies plays a major role in the assessment of lymphadenopathy and staging of metastatic disease. Radiologists routinely measure the nodal size in order to distinguish benign from malignant nodes, which require subsequent cancer staging. However, identification of lymph nodes is a cumbersome task due to their myriad appearances in mpMRI studies. Multiple sequences are acquired in mpMRI studies, including T2 fat suppressed (T2FS) and diffusion weighted imaging (DWI) sequences among others; consequently, the sizing of LNs is rendered challenging due to the variety of signal intensities in these sequences. Furthermore, radiologists can miss potentially metastatic LNs during a busy clinical day. To lighten these imaging and workflow challenges, we propose a computer-aided detection (CAD) pipeline to detect both benign and malignant LNs in the body for their subsequent measurement. We employed the recently proposed Dynamic Head (DyHead) neural network to detect LNs in mpMRI studies that were acquired using a variety of scanners and exam protocols. The T2FS and DWI series were co-registered, and a selective augmentation technique called Intra-Label LISA (ILL) was used to blend the two volumes with the interpolation factor drawn from a Beta distribution. In this way, ILL diversified the samples that the model encountered during the training phase, while the requirement for both sequences to be present at test time was nullified. Our results showed a mean average precision (mAP) of 53.5% and a sensitivity of ∼78% with ILL at 4 FP/vol. This corresponded to an improvement of ≥10% in mAP and ≥12% in sensitivity at 4FP (p ¡ 0.05) respectively over current LN detection approaches evaluated on the same dataset. We also established the out-of-distribution robustness of the DyHead model by training it on data acquired by a Siemens Aera scanner and testing it on data from the Siemens Verio, Siemens Biograph mMR, and Philips Achieva scanners. Our pilot work represents an important first step towards automated detection, segmentation, and classification of lymph nodes in mpMRI.

Universal detection and segmentation of lymph nodes in multi-parametric MRI.

PURPOSE: Reliable measurement of lymph nodes (LNs) in multi-parametric MRI (mpMRI) studies of the body plays a major role in the assessment of lymphadenopathy and staging of metastatic disease. Previous approaches do not adequately exploit the complementary sequences in mpMRI to universally detect and segment lymph nodes, and they have shown fairly limited performance. METHODS: We propose a computer-aided detection and segmentation pipeline to leverage the T2 fat-suppressed (T2FS) and diffusion-weighted imaging (DWI) series from a mpMRI study. The T2FS and DWI series in 38 studies (38 patients) were co-registered and blended together using a selective data augmentation technique, such that traits of both series were visible in the same volume. A mask RCNN model was subsequently trained for universal detection and segmentation of 3D LNs. RESULTS: Experiments on 18 test mpMRI studies revealed that the proposed pipeline achieved a precision of [Formula: see text]%, sensitivity of [Formula: see text]% at 4 false positives (FP) per volume, and dice score of [Formula: see text]%. This represented an improvement of [Formula: see text]% in precision, [Formula: see text]% in sensitivity at 4 FP/volume, and [Formula: see text]% in dice score, respectively, over current approaches evaluated on the same dataset. CONCLUSION: Our pipeline universally detected and segmented both metastatic and non-metastatic nodes in mpMRI studies. At test time, the input data used by the trained model could either be the T2FS series alone or a blend of co-registered T2FS and DWI series. Contrary to prior work, this eliminated the reliance on both the T2FS and DWI series in a mpMRI study.

Universal lymph node detection in T2 MRI using neural networks.

PURPOSE: Identification of lymph nodes (LNs) that are suspicious for metastasis in T2 Magnetic Resonance Imaging (MRI) is critical for assessment of lymphadenopathy. Prior work on LN detection has been limited to specific anatomical regions of the body (pelvis, rectum). Therefore, an approach to universally detect both benign and metastatic nodes in T2 MRI studies of the body is highly desirable. METHODS: We developed a Computer Aided Detection (CAD) pipeline to universally identify LN in T2 MRI. First, we trained various neural networks for detecting LN: Faster RCNN with and without Hard Negative Example Mining (HNEM), FCOS, FoveaBox, VFNet, and Detection Transformer (DETR). Next, we show that VFNet with Adaptive Training Sample Selection (ATSS) outperformed Faster RCNN with HNEM. Finally, we ensembled models that surpassed a 45% mAP threshold. RESULTS: Experiments on 122 test studies revealed that VFNet achieved a 51.1% mAP and 78.7% recall at 4 false positives (FP) per volume, while the one-stage model ensemble achieved a mAP of 52.3% and sensitivity of 78.7% at 4FP. We found that VFNet and the one-stage model ensemble can be interchangeably used in the CAD pipeline. CONCLUSION: Our CAD pipeline universally detected both benign and metastatic nodes in T2 MRI studies, resulting in a sensitivity improvement of [Formula: see text]14% over the current LN detection approaches (sensitivity of 78.7% at 4 FP vs. 64.6% at 5 FP per volume).

Reticular Pseudodrusen: The Third Macular Risk Feature for Progression to Late Age-Related Macular Degeneration: Age-Related Eye Disease Study 2 Report 30.

PURPOSE: To analyze reticular pseudodrusen (RPD) as an independent risk factor for progression to late age-related macular degeneration (AMD), alongside traditional macular risk factors (soft drusen and pigmentary abnormalities) considered simultaneously. DESIGN: Post hoc analysis of 2 clinical trial cohorts: Age-Related Eye Disease Study (AREDS) and AREDS2. PARTICIPANTS: Eyes with no late AMD at baseline in AREDS (6959 eyes, 3780 participants) and AREDS2 (3355 eyes, 2056 participants). METHODS: Color fundus photographs (CFPs) from annual visits were graded for soft drusen, pigmentary abnormalities, and late AMD. Presence of RPD was from grading of fundus autofluorescence images (AREDS2) and deep learning grading of CFPs (AREDS). Proportional hazards regression analyses were performed, considering AREDS AMD severity scales (modified simplified severity scale [person] and 9-step scale [eye]) and RPD presence simultaneously. MAIN OUTCOME MEASURES: Progression to late AMD, geographic atrophy (GA), and neovascular AMD. RESULTS: In AREDS, for late AMD analyses by person, in a model considering the simplified severity scale simultaneously, RPD presence was associated with a higher risk of progression: hazard ratio (HR), 2.15 (95% confidence interval [CI], 1.75-2.64). However, the risk associated with RPD presence differed at different severity scale levels: HR, 3.23 (95% CI, 1.60-6.51), HR, 3.81 (95% CI, 2.38-6.10), HR, 2.28 (95% CI, 1.59-3.27), and HR, 1.64 (95% CI, 1.20-2.24), at levels 0-1, 2, 3, and 4, respectively. Considering the 9-step scale (by eye), RPD presence was associated with higher risk: HR, 2.54 (95% CI, 2.07-3.13). The HRs were 5.11 (95% CI, 3.93-6.66) at levels 1-6 and 1.78 (95% CI, 1.43-2.22) at levels 7 and 8. In AREDS2, by person, RPD presence was not associated with higher risk: HR, 1.18 (95% CI, 0.90-1.56); by eye, it was HR, 1.57 (95% CI, 1.31-1.89). In both cohorts, RPD presence carried a higher risk for GA than neovascular AMD. CONCLUSIONS: Reticular pseudodrusen represent an important risk factor for progression to late AMD, particularly GA. However, the added risk varies markedly by severity level, with highly increased risk at lower/moderate levels and less increased risk at higher levels. Reticular pseudodrusen status should be included in updated AMD classification systems, risk calculators, and clinical trials.

DDX04+ Stem Cells in the Ovaries of Postmenopausal Women: Existence and Differentiation Potential.

Ovarian aging is a pacemaker with multiple organ dysfunction. Recently, stem cells with the ability to generate new oocytes have been identified, which provides the possibility of stem cell therapy for ovarian aging. Several studies have revealed the existence of stem cells in the human postmenopausal ovary. In this study, we describe a new method using magnetic-activated cell sorting combined with differential adhesion to isolate DDX4+ stem cells from ovaries of postmenopausal women and show that the cells exhibit similar gene expression profiles and growth characteristics with primitive germ cells. Furthermore, the DDX4+ stem cells could enter the meiosis stage and differentiation into oocytes. The RNA-seq data of the differentiated oocytes shows that mitochondrial metabolism may play an important role in the oogenesis process of the DDX4+ stem cells. Through using the human ovarian cortical fragments transplantation model, we indicated that the GFP-DDX4+ stem cells differentiated into some GFP positive oocyte-like structure in vivo. Our study provided a new method for the isolation of DDX4+ stem cells from the ovaries of postmenopausal women and confirmed the ability of these stem cells to differentiate into oocytes.

Machine Learning Approach to Facilitate Knowledge Synthesis at the Intersection of Liver Cancer, Epidemiology, and Health Disparities Research.

PURPOSE: Liver cancer is a global challenge, and disparities exist across multiple domains and throughout the disease continuum. However, liver cancer's global epidemiology and etiology are shifting, and the literature is rapidly evolving, presenting a challenge to the synthesis of knowledge needed to identify areas of research needs and to develop research agendas focusing on disparities. Machine learning (ML) techniques can be used to semiautomate the literature review process and improve efficiency. In this study, we detail our approach and provide practical benchmarks for the development of a ML approach to classify literature and extract data at the intersection of three fields: liver cancer, health disparities, and epidemiology. METHODS: We performed a six-phase process including: training (I), validating (II), confirming (III), and performing error analysis (IV) for a ML classifier. We then developed an extraction model (V) and applied it (VI) to the liver cancer literature identified through PubMed. We present precision, recall, F1, and accuracy metrics for the classifier and extraction models as appropriate for each phase of the process. We also provide the results for the application of our extraction model. RESULTS: With limited training data, we achieved a high degree of accuracy for both our classifier and for the extraction model for liver cancer disparities research literature performed using epidemiologic methods. The disparities concept was the most challenging to accurately classify, and concepts that appeared infrequently in our data set were the most difficult to extract. CONCLUSION: We provide a roadmap for using ML to classify and extract comprehensive information on multidisciplinary literature. Our technique can be adapted and modified for other cancers or diseases where disparities persist.

Absolute film separation of dyes/salts and emulsions with a superhigh water permeance through graded nanofluidic channels.

The development of multifunctional films with a high permeability has been of great concern for effective separation of complex aqueous contaminants, especially in the face of zero or near-zero release regulations. Inspired by the natural structure of sandy soils, polydopamine-wrapped/connected polypyrrole sub-micron spheres (PPSM) were closely packed onto a polypyrrole-coated bacterial cellulose (PBC) support, by which a new two-layered PBC/PPSM composite film formed with graded nanofluidic channels. Interestingly, after being soaked in complex water environments of ethanol, acids, bases, heat, cold and high salinity, or else bended/folded for more than 10 times, the structure and performance of this film still stayed the same, validating its high structural stability and flexibility. Even in a high salinity environment over seawater, this PBC/PPSM film exhibits a dye-separation capacity of almost 100% with a surprisingly superhigh water permeance over one thousand L h-1 m-2 bar-1, one or two magnitudes higher than that of the related films reported in the literature. Meanwhile, the ability for effective oil-water-separation was also validated. Besides the superhydrophilicity and underwater superoleophobicity, the synapse-like-structure-induced graded nanofluidic channels are also proposed to play a key role for rendering such an outstandingly comprehensive performance of the film by greatly overcoming fluid resistance and reducing permeation viscosity.

Woolly hair nevus caused by somatic mutation and Costello syndrome caused by germline mutation in HRAS: Consider parental mosaicism in prenatal counseling.

Germline mutations in HRAS cause Costello syndrome (CS), while mosaic mutations in HRAS show a variability of phenotypes, ranging from mild features such as keratinocytic epidermal nevus (KEN), sebaceous nevus (SN), woolly hair nevus (WHN) with KEN, to severe manifestations of CS with cutis laxa. We report two individuals. The first was a 2-year-old boy with woolly hair nevus (WHN) without any other cutaneous involvement, in whom somatic HRAS mutation (c.34G>A; p.Gly12Ser) was identified in his affected scalp and hair follicle specimens. This is the first reported WHN type 1 (no cutaneous involvement) patient caused by somatic HRAS mutation. The other individual was a 12-year-old girl with CS caused by germline HRAS mutation (c.34G>A), that manifested with coarse face, palmoplantar keratoderma, deep palmar and plantar creases, hyperpigmented patches, asymmetry and deformity of lower limbs, atopic dermatitis, as well as mental retardation. Of note, a linear hyperpigmented plaque was observed in her father's lumbosacral region. Although the father refused to provide semen and skin tissue for further examination, this reminds us of possible mosaicism in parents of individuals with germline de novo HRAS mutation and underlines the importance of parental evaluation for prenatal counseling.

Free-standing hybrid film for separation of dye pollutant with self-cleaning ability under visible light.

The development of low cost and environmental-friendly materials has long been an ambition for effective removal of dye pollutants in complex water environments. In this study, a free-standing separation film of bacterial cellulose reinforced/functionalized by graphitic phase carbon nitride is developed by a facile suction filtration strategy, of which the former is precoated by polypyrrole, and the latter is pre-doped by oxygen to endow the as-obtained film an enhanced photocatalytic performance and self-cleaning ability. The as-obtained film exhibits a high tensile stress of 51.8 ± 1.1 MPa, and a high resistance to cold, heat, acid and alkali. For typical dyes of methylene blue and rhodamine B, a high dye rejection rate of 99.9% at 138 L/m2•h•bar feed flux is obtained by the as-obtained film. Even at a salt concentration higher than 5%, it still maintained high dye removal rates and achieves effective separation of dye and salt. Simultaneously, a high dye photocatalytic degradation of the composite films rates up to 98% in only 90 min, and a high self-cleaning ability demonstrated by recovery of flux after light treatment in cyclic tests. The density functional theory calculation validates the beneficial effects of improved light response range and separated photogenerated electron/holes for the effective degradation of dyes by oxygen-doped carbon nitride coupled with one-dimensional polypyrrole chains. Overall, this study proposes a new direction for the separation of dye pollutants with a high visible-light self-cleaning capacity by structural tailoring of bacterial cellulose with carbon nitride.

LitSuggest: a web-based system for literature recommendation and curation using machine learning.

Searching and reading relevant literature is a routine practice in biomedical research. However, it is challenging for a user to design optimal search queries using all the keywords related to a given topic. As such, existing search systems such as PubMed often return suboptimal results. Several computational methods have been proposed as an effective alternative to keyword-based query methods for literature recommendation. However, those methods require specialized knowledge in machine learning and natural language processing, which can make them difficult for biologists to utilize. In this paper, we propose LitSuggest, a web server that provides an all-in-one literature recommendation and curation service to help biomedical researchers stay up to date with scientific literature. LitSuggest combines advanced machine learning techniques for suggesting relevant PubMed articles with high accuracy. In addition to innovative text-processing methods, LitSuggest offers multiple advantages over existing tools. First, LitSuggest allows users to curate, organize, and download classification results in a single interface. Second, users can easily fine-tune LitSuggest results by updating the training corpus. Third, results can be readily shared, enabling collaborative analysis and curation of scientific literature. Finally, LitSuggest provides an automated personalized weekly digest of newly published articles for each user's project. LitSuggest is publicly available at https://www.ncbi.nlm.nih.gov/research/litsuggest.

THERAPY OF ENDOCRINE DISEASE: Novel protection and treatment strategies for chemotherapy-associated ovarian damage.

Fertility and ovarian protection against chemotherapy-associated ovarian damage has formed a new field called oncofertility, which is driven by the pursuit of fertility protection as well as good life quality for numerous female cancer survivors. However, the choice of fertility and ovarian protection method is a difficult problem during chemotherapy and there is no uniform guideline at present. To alleviate ovarian toxicity caused by anticancer drugs, effective methods combined with an individualized treatment plan that integrates an optimal strategy for preserving and restoring reproductive function should be offered from well-established to experimental stages before, during, and after chemotherapy. Although embryo, oocyte, and ovarian tissue cryopreservation are the major methods that have been proven effective and feasible for fertility protection, they are also subject to many limitations. Therefore, this paper mainly discusses the future potential methods and corresponding mechanisms for fertility protection in chemotherapy-associated ovarian damage.

Recent advances of automated methods for searching and extracting genomic variant information from biomedical literature.

MOTIVATION: To obtain key information for personalized medicine and cancer research, clinicians and researchers in the biomedical field are in great need of searching genomic variant information from the biomedical literature now than ever before. Due to the various written forms of genomic variants, however, it is difficult to locate the right information from the literature when using a general literature search system. To address the difficulty of locating genomic variant information from the literature, researchers have suggested various solutions based on automated literature-mining techniques. There is, however, no study for summarizing and comparing existing tools for genomic variant literature mining in terms of how to search easily for information in the literature on genomic variants. RESULTS: In this article, we systematically compared currently available genomic variant recognition and normalization tools as well as the literature search engines that adopted these literature-mining techniques. First, we explain the problems that are caused by the use of non-standard formats of genomic variants in the PubMed literature by considering examples from the literature and show the prevalence of the problem. Second, we review literature-mining tools that address the problem by recognizing and normalizing the various forms of genomic variants in the literature and systematically compare them. Third, we present and compare existing literature search engines that are designed for a genomic variant search by using the literature-mining techniques. We expect this work to be helpful for researchers who seek information about genomic variants from the literature, developers who integrate genomic variant information from the literature and beyond.

Anticancer effects of brusatol in nasopharyngeal carcinoma through suppression of the Akt/mTOR signaling pathway.

PURPOSE: Brusatol, a natural quassinoid that is isolated from a traditional Chinese herbal medicine known as Bruceae Fructus, possesses biological activity in various types of human cancers, but its effects in nasopharyngeal carcinoma (NPC) have not been reported. This study aimed to explore the effect and molecular mechanism of brusatol in NPC in vivo and in vitro. METHODS: The antiproliferative effect of brusatol was assessed by MTT and colony formation assays. Apoptosis was determined by flow cytometry. The expression of mitochondrial apoptosis, cell cycle arrest, and Akt/mTOR pathway proteins were determined by western blot analysis. Further in vivo confirmation was performed in a nude mouse model. RESULTS: Brusatol showed antiproliferative activity against four human NPC cell lines (CNE-1, CNE-2, 5-8F, and 6-10B) in a dose-dependent manner. This antiproliferative effect was accompanied by mitochondrial apoptosis and cell cycle arrest through the modulation of several key molecular targets, such as Bcl-xl, Bcl-2, Bad, Bax, PARP, Caspase-9, Caspase-7, Caspase-3, Cdc25c, Cyclin B1, Cdc2 p34, and Cyclin D1. In addition, we found that brusatol inhibited the activation of Akt, mTOR, 4EBP1, and S6K, suggesting that the Akt/mTOR pathway is a key underlying mechanism by which brusatol inhibits growth and promotes apoptosis. Further in vivo nude mouse models proved that brusatol significantly inhibited the growth of CNE-1 xenografts with no significant toxicity. CONCLUSIONS: These observations indicate that brusatol is a promising antitumor drug candidate or a supplement to current chemotherapeutic therapies to treat NPC.

Tracking human genes along the translational continuum.

Understanding the drivers of research on human genes is a critical component to success of translation efforts of genomics into medicine and public health. Using publicly available curated online databases we sought to identify specific genes that are featured in translational genetic research in comparison to all genomics research publications. Articles in the CDC's Public Health Genomics and Precision Health Knowledge Base were stratified into studies that have moved beyond basic research to population and clinical epidemiologic studies (T1: clinical and population human genome epidemiology research), and studies that evaluate, implement, and assess impact of genes in clinical and public health areas (T2+: beyond bench to bedside). We examined gene counts and numbers of publications within these phases of translation in comparison to all genes from Medline. We are able to highlight those genes that are moving from basic research to clinical and public health translational research, namely in cancer and a few genetic diseases with high penetrance and clinical actionability. Identifying human genes of translational value is an important step towards determining an evidence-based trajectory of the human genome in clinical and public health practice over time.

Coinheritance of generalized pustular psoriasis and familial Behçet-like autoinflammatory syndrome with variants in IL36RN and TNFAIP3 in the heterozygous state.

Generalized pustular psoriasis (GPP) is now known to be caused by biallelic variants in IL36RN and monoallelic variants in CARD14 and AP1S3. The presence of a modifier locus or oligogenic inheritance have been hypothesized. We report on a patient with a unique coinheritance of pathogenic variants in IL36RN (c.115+6T>C) and TNFAIP3 (c.547C>T, p.R183*) causing the genetic entities GPP and familial Behçet-like autoinflammatory syndrome (AISBL). The heterozygous variant in IL36RN identified by Sanger sequencing was inherited from his unaffected father, while the heterozygous variant in TNFAIP3 was detected by whole-exome sequencing and was also identified in the patient's AISBL-affected maternal relatives. Further functional studies are required to research whether the variant of TNFAIP3 plays a part in the development of GPP or simply causes the Behçet's disease phenotype. However, our data suggest that whole-exome sequencing for the heterozygous carrier of the IL36RN gene in GPP be used to find the potential second genetic locus.

Ankyloblepharon-ectodermal dysplasia-clefting syndrome misdiagnosed as epidermolysis bullosa and congenital ichthyosiform erythroderma: Case report and review of published work.

A Chinese female infant presented with ectodermal dysplasia, cleft palate and severe skin erosions at birth. Although all the typical clinical features of ankyloblepharon-ectodermal dysplasia-clefting (AEC) syndrome were present, the ankyloblepharon was not very marked. We misdiagnosed epidermolysis bullosa and congenital ichthyosiform erythroderma at first and confirmed the diagnosis of AEC syndrome only when she presented with the typical clinical manifestation of recurrent infected scalp erosions at 1 year of age. Mutation analysis of exon 13 of the p63 gene revealed a missense mutation Ile482Thr (c.1445T>C) in the sterile alpha motive domain. In this work we review the clinical features, differential diagnosis and prognosis in AEC syndrome.

Resveratrol alleviates chemotherapy-induced oogonial stem cell apoptosis and ovarian aging in mice.

Chemotherapy-induced ovarian aging not only increases the risk for early menopause-related complications but also results in infertility in young female cancer survivors. Oogonial stem cells have the ability to generate new oocytes and thus provide new opportunities for treating ovarian aging and female infertility. Resveratrol (3,5,4'-trihydroxy-trans-stilbene) is a natural phenol derived from plants, that has been shown to have positive effects on longevity and redox flow in lipid metabolism and a preventive function against certain tumors. To evaluate whether resveratrol could promote the repair of oogonial stem cells damage in a busulfan/cyclophosphamide (Bu/Cy)-induced accelerated ovarian aging model, female mice were administered 30 and 100 mg/kg/d resveratrol through a gavage for 2 weeks. We demonstrated that resveratrol (30 mg/kg/d) relieved oogonial stem cells loss and showed an attenuating effect on Bu/Cy-induced oxidative apoptosis in mouse ovaries, which may be attributed to the attenuation of oxidative levels in ovaries. Additionally, we also showed that Res exerted a dose-dependent effect on oogonial stem cells and attenuated H2O2-induced cytotoxicity and oxidative stress injury by activating Nrf2 in vitro. Therefore, resveratrol could be of a potential therapeutic drug used to prevent chemotherapy-induced ovarian aging.

Enhanced Breakthrough Efficiency by a Chemically Stable Porous Coordination Polymer with Optimized Nanochannel.

High separation efficiency is very important for process of pressure swing adsorption (PSA) in the industry. Herein, we propose a fine design of chemically stable porous coordination polymers (PCPs) with optimized nanochannel by strategy of inserting and shifting shortest alkyl group on T-shaped ligand. Remarkably, the synergistic effect of optimized nanochannel, unique crystal morphology and fitted channel enable sharply enhanced breakthrough efficiency of C2H6/4/CH4, 1.17 or 0.77 g of CH4 can be separated from corresponding dual mixtures (2/8, v/v) by 1 g of NTU-25 at 273 K, which was further validated and understood by controlled experiments and density functional theory (DFT) computations.

Enrichment of Female Germline Stem Cells from Mouse Ovaries Using the Differential Adhesion Method.

BACKGROUND/AIMS: The isolation and establishment of female germline stem cells (FGSCs) is controversial because of questions regarding the reliability and stability of the isolation method using antibody targeting mouse vasa homologue (MVH), and the molecular mechanism of FGSCs self-renewal remains unclear. Thus, there needs to be a simple and reliable method for sorting FGSCs to study them. METHODS: We applied the differential adhesion method to enrich FGSCs (DA-FGSCs) from mouse ovaries. Through four rounds of purification and 7-9 subsequent passages, DA-FGSC lines were established. In addition, we assessed the role of the phosphoinositide-3 kinase (PI3K)-AKT pathway in regulating FGSC self-renewal. RESULTS: The obtained DA-FGSCs spontaneously differentiated into oocyte-like cells in vitro and formed functional eggs in vivo that were fertilized and produced healthy offspring. AKT was rapidly phosphorylated when the proliferation rate of FGSCs increased after 10 passages, and the addition of a chemical PI3K inhibitor prevented FGSCs self-renewal. Furthermore, over-expression of AKT-induced proliferation and differentiation of FGSCs, c-Myc, Oct-4 and Gdf-9 levels were increased. CONCLUSIONS: The differential adhesion method provides a more feasible approach and is an easier procedure to establish FGSC lines than traditional methods. The AKT pathway plays an important role in regulation of the proliferation and maintenance of FGSCs. These findings could help promote stem cell studies and provide a better understanding of causes of ovarian infertility, thereby providing potential treatments for infertility.