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Breast and prostate cancer research to date has largely been predicated on the use of cell lines in vitro or in vivo. These limitations have led to the development of more clinically relevant models, such as organoids or murine xenografts that utilize patient‐derived material; however, issues related to low take rate, long duration of establishment, and the associated costs constrain use of these models. This study demonstrates that ex vivo culture of freshly resected breast and prostate tumor specimens obtained from surgery, termed patient‐derived explants (PDEs), provides a high‐throughput and cost‐effective model that retains the native tissue architecture, microenvironment, cell viability, and key oncogenic drivers. The PDE model provides a unique approach for direct evaluation of drug responses on an individual patient's tumor, which is amenable to analysis using contemporary genomic technologies. The ability to rapidly evaluate drug efficacy in patient‐derived material has high potential to facilitate implementation of personalized medicine approaches. This study describes an innovative preclinical model that has significant potential to accelerate translational cancer research outcomes. Breast and prostate tumors were cultured as patient‐derived explants (PDE), in a way that sustains the native tissue architecture, microenvironment, cell viability, and steroid receptor signaling. PDEs were responsive to hormones, therapeutic agents, and shRNA while in culture, allowing direct comparison of treatments within an individual's tumor using high‐throughput molecular approaches, including cistrome profiling.

Hepsin, a type II transmembrane serine protease, is commonly overexpressed in prostate and breast cancer. The hepsin protein is stabilized by the Ras‐MAPK pathway, and, downstream, this protease regulates the degradation of extracellular matrix components and activates growth factor pathways, such as the hepatocyte growth factor (HGF) and transforming growth factor beta (TGFβ) pathway. However, how exactly active hepsin promotes cell proliferation machinery to sustain tumor growth is not fully understood. Here, we show that genetic deletion of the gene encoding hepsin (Hpn) in a WAP‐Myc model of aggressive MYC‐driven breast cancer inhibits tumor growth in the primary syngrafted sites and the growth of disseminated tumors in the lungs. The suppression of tumor growth upon loss of hepsin was accompanied by downregulation of TGFβ and EGFR signaling together with a reduction in epidermal growth factor receptor (EGFR) protein levels. We further demonstrate in 3D cultures of patient‐derived breast cancer explants that both basal TGFβ signaling and EGFR protein expression are inhibited by neutralizing antibodies or small‐molecule inhibitors of hepsin. The study demonstrates a role for hepsin as a regulator of cell proliferation and tumor growth through TGFβ and EGFR pathways, warranting consideration of hepsin as a potential indirect upstream target for therapeutic inhibition of TGFβ and EGFR pathways in cancer. Serine protease hepsin, which is commonly overexpressed in prostate, ovarian, and breast cancers, promotes cancer progression with yet unclear mechanisms. We show with breast cancer patient‐derived explant cultures (PDECs), Myc‐driven mouse mammary tumors, and cell culture models that ablation of hepsin activity inhibits tumor growth and cell proliferation through loss of TGFβ and EGFR signaling in breast cancer context.

[This corrects the article DOI: 10.1371/journal.pone.0181962.].

Musculoskeletal injuries, including tendinopathies, present a significant clinical burden for aging populations. While the biological drivers of age‐related declines in tendon function are poorly understood, it is well accepted that dysregulation of extracellular matrix (ECM) remodeling plays a role in chronic tendon degeneration. Senescent cells, which have been associated with multiple degenerative pathologies in musculoskeletal tissues, secrete a highly pro‐inflammatory senescence‐associated secretory phenotype (SASP) that has potential to promote ECM breakdown. However, the role of senescent cells in the dysregulation of tendon ECM homeostasis is largely unknown. To assess this directly, we developed an in vitro model of induced cellular senescence in murine tendon explants. This novel technique enables us to study the isolated interactions of senescent cells and their native ECM without interference from age‐related systemic changes. We document multiple biomarkers of cellular senescence in induced tendon explants including cell cycle arrest, apoptosis resistance, and sustained inflammatory responses. We then utilize this in vitro senescence model to compare the ECM remodeling response of young, naturally aged, and induced‐senescent tendons to an altered mechanical stimulus. We found that both senescence and aging independently led to alterations in ECM‐related gene expression, reductions in protein synthesis, and tissue compositional changes. Furthermore, MMP activity was sustained, thus shifting the remodeling balance of aged and induced‐senescent tissues towards degradation over production. Together, this demonstrates that cellular senescence plays a role in the altered mechano‐response of aged tendons and likely contributes to poor clinical outcomes in aging populations. We developed a novel in vitro model of induced cellular senescence in murine tendon explants and then utilized this model to compare ECM remodeling of young, aged, and induced‐senescent tendons to an altered mechanical stimulus. Both senescence and aging led to reduced protein synthesis with sustained MMP activity, thus shifting the remodeling balance towards degradation over production. This implicates senescence as a key player in altered mechanobiology of aged tendons and a likely contributor to poor clinical outcomes.

High transformation efficiency is a prerequisite for study of gene function and molecular breeding. -mediated transformation is a preferred method in many plants. However, the transformation efficiency in soybean is still low. The objective of this study is to optimize -mediated transformation in soybean by improving the infection efficiency of and regeneration efficiency of explants. Firstly, four factors affecting infection efficiency were investigated by estimation of the rate of GUS transient expression in soybean cotyledonary explants, including concentrations, soybean explants, suspension medium, and co-cultivation time. The results showed that an infection efficiency of over 96% was achieved by collecting the at a concentration of OD = 0.6, then using an suspension medium containing 154.2 mg/L dithiothreitol to infect the half-seed cotyledonary explants (from mature seeds imbibed for 1 day), and co-cultured them for 5 days. The infection efficiencies for soybean varieties Jack Purple and Tianlong 1 were higher than the other six varieties. Secondly, the rates of shoot elongation were compared among six different concentration combinations of gibberellic acid (GA ) and indole-3-acetic acid (IAA). The shoot elongation rate of 34 and 26% was achieved when using the combination of 1.0 mg/L GA and 0.1 mg/L IAA for Jack Purple and Tianlong 1, respectively. This rate was higher than the other five concentration combinations of GA and IAA, with an 18 and 11% increase over the original laboratory protocol (a combination of 0.5 mg/L GA and 0.1 mg/L IAA), respectively. The transformation efficiency was 7 and 10% for Jack Purple and Tianlong 1 at this optimized hormone concentration combination, respectively, which was 2 and 6% higher than the original protocol, respectively. Finally, GUS histochemical staining, PCR, herbicide (glufosinate) painting, and QuickStix Kit for Liberty Link ( ) were used to verify the positive transgenic plants, and absolute quantification PCR confirmed the exogenous gene existed as one to three copies in the soybean genome. This study provides an improved protocol for -mediated transformation in soybean and a useful reference to improve the transformation efficiency in other plant species.

Catasetum integerrimum is an orchid used in Mexican traditional medicine, which is overexploited due to its ornamental and medicinal properties. In this scenario, conservation strategies are mandatory to protect wild populations from this valuable orchid. The aim of the present study was to establish a micropropagation protocol by direct organogenesis, as a tool for Catasetum integerrimum production and conservation. Seeds from a mature capsule were cultivated in a Murashige and Skoog basal medium. Then, 6-mo-old vitroplants were transplanted into treatments fortified with 6-bencilaminopurin (BAP) or indole-3-acetic acid (IAA) to induce plant development. Treatment fortified with 2.5 mg L⁻¹ BAP in combination with 5.0 mg L⁻¹ IAA developed 5.73 ± 0.45 shoots per explant and 5.84 ± 0.48 leaves per shoot. For the rooting process, the best treatment was 2.5 mg L⁻¹ IAA, which developed 11.20 ± 0.28 roots with a 13.20 ± 0.28-cm root length. C. integerrimum vitroplants were used as a source of leaves, roots, and pseudobulbs for the induction of direct organogenesis. Leaf explants did not induce any morphogenic response. Pseudobulb explants had the best response when developing 1 shoot per explant with 5.50 ± 0.18 leaves per shoot and 4.37 ± 0.37 roots per explant with a 4.88 ± 0.20-cm root length. For the acclimatization stage, a survival rate of over 90% was achieved. The results obtained prove the efficacy of the micropropagation method, which contributes significantly to the conservation of this orchid, preserving its wild populations from indiscriminate collection.

Abstract Study Design Observational study using insurance claims. Objective To quantify opioid usage leading up to spinal cord stimulation (SCS) and the potential impact on outcomes of SCS. Setting SCS is an interventional therapy that often follows opioid usage in the care continuum for chronic pain. Methods This study identified SCS patients using the Truven Health MarketScan databases from January 2010 to December 2014. The index event was the first occurrence of a permanent SCS implant. Indicators of opioid usage at implant were daily morphine equivalent dose (MED), number of unique pain drug classes, and diagnosis code for opioid abuse. System explant was used as a measure of ineffective SCS therapy. Multivariate logistic regression was used to analyze the effect of pre-implant medications on explants. Results A total of 5,476 patients (56 ± 14 years; 60% female) were included. SCS system removal occurred in 390 patients (7.1%) in the year after implant. Number of drug classes (odds ratio [OR] = 1.11, P = 0.007) and MED level (5–90 vs < 5 mg/d: OR = 1.32, P = 0.043; ≥90 vs < 5 mg/d: OR = 1.57, P = 0.005) were independently predictive of system explant. Over the year before implant, MED increased in 54% (stayed the same in 21%, decreased in 25%) of patients who continued with SCS and increased in 53% (stayed the same in 20%, decreased in 27%) of explant patients (P = 0.772). Over the year after implant, significantly more patients with continued SCS had an MED decrease (47%) or stayed the same (23%) than before (P < 0.001). Conclusions Chronic pain patients receive escalating opioid dosage prior to SCS implant, and high-dose opioid usage is associated with an increased risk of explant. Neuromodulation can stabilize or decrease opioid usage. Earlier consideration of SCS before escalated opioid usage has the potential to improve outcomes in complex chronic pain.

Chrysanthemum is a flowering plant grown worldwide and is one of the most popular ornamental plants. Chrysanthemums are usually cultivated using root suckers and shoot cuttings. This conventional technique is relatively slow. In addition, as cuttings are gained regularly from mother plants, there is a chance of viral infection and degeneration, which raises the production cost. The hurdles mentioned above have been managed by applying in vitro propagation techniques, which can enhance reproduction rates through in vitro culture and use very small explants, which are impossible with the conventional approach. Usually, it is difficult to get true-to-type plants as the parents with good quality, but clonal propagation of a designated elite species makes it possible. Hence, this review highlights recent studies of the in vitro propagation of Chrysanthemum included; the appropriate explant sources, medium compositions, alternative disinfection of culture media, plant growth regulators (PGRs), different mutagenesis applications, acclimatization efficiency, and alternative light sources to overcome the shortcomings of conventional propagation techniques.

is an obligate human pathogen that causes mucosal surface infections of male and female reproductive tracts, pharynx, rectum, and conjunctiva. Asymptomatic or unnoticed infections in the lower reproductive tract of women can lead to serious, long-term consequences if these infections ascend into the fallopian tube. The damage caused by gonococcal infection and the subsequent inflammatory response produce the condition known as pelvic inflammatory disease (PID). Infection can lead to tubal scarring, occlusion of the oviduct, and loss of critical ciliated cells. Consequences of the damage sustained on the fallopian tube epithelium include increased risk of ectopic pregnancy and tubal-factor infertility. Additionally, the resolution of infection can produce new adhesions between internal tissues, which can tear and reform, producing chronic pelvic pain. As a bacterium adapted to life in a human host, the gonococcus presents a challenge to the development of model systems for probing host-microbe interactions. Advances in small-animal models have yielded previously unattainable data on systemic immune responses, but the specificity of for many known (and unknown) host targets remains a constant hurdle. Infections of human volunteers are possible, though they present ethical and logistical challenges, and are necessarily limited to males due to the risk of severe complications in women. It is routine, however, that normal, healthy fallopian tubes are removed in the course of different gynecological surgeries (namely hysterectomy), making the very tissue most consequentially damaged during ascending gonococcal infection available for laboratory research. The study of fallopian tube organ cultures has allowed the opportunity to observe gonococcal biology and immune responses in a complex, multi-layered tissue from a natural host. Forty-five years since the first published example of human fallopian tube being infected with , we review what modeling infections in human tissue explants has taught us about the gonococcus, what we have learned about the defenses mounted by the human host in the upper female reproductive tract, what other fields have taught us about ciliated and non-ciliated cell development, and ultimately offer suggestions regarding the next generation of model systems to help expand our ability to study gonococcal pathogenesis.