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The aim of this study was to measure umbilical blood androgen concentrations in a birth cohort using a highly specific liquid chromatography-tandem mass spectrometry (LC-MS/MS) assay and assesses the effects of sex, labor, and gestational age on fetal androgen levels at birth. We performed a prospective cohort study of androgen concentrations in mixed arterial and venous umbilical cord serum from 803 unselected singleton pregnancies from a general obstetric population in Western Australia. Total testosterone (TT), Δ4-androstenedione, and dehydroepiandrosterone were extracted from archived cord serum samples and measured using LC-MS/MS. SHBG was measured by ELISA; free testosterone (FT) and bioavailable testosterone (BioT) values were also calculated. Median values for all three androgens were generally lower than previously published values. Levels of TT, FT, BioT, and SHBG were significantly higher in male verses female neonates (P<0.0001), while dehydroepiandrosterone levels were higher in females (P<0.0001). Labor was associated with a significant (∼15-26%) decrease in median cord blood TT and FT levels (both sexes combined), but a modest (∼16-31%) increase in SHBG, Δ4-androstenedione, and dehydroepiandrosterone concentrations. TT and FT were significantly negatively correlated with gestational age at delivery, while SHBG, Δ4-androstenedione, and dehydroepiandrosterone were positively correlated. Antenatal glucocorticoid administration also had a significant effect in the multiple regression models. This is the first study to report umbilical cord androgen levels in a large unselected population of neonates using LC-MS/MS. Our findings suggest that previous studies have over-estimated cord androgen levels, and that fetal, maternal, and obstetric factors influence cord androgen levels differentially. Caution should be exercised when interpreting previously-published data that have not taken all of these factors into account.

Abstract Disease progression and relapse in multiple myeloma is dependent on the ability of the multiple myeloma plasma cells (PC) to reenter the circulation and disseminate throughout the bone marrow. Increased bone marrow hypoxia is associated with increased recirculation of multiple myeloma PCs. Accordingly, we hypothesized that during chronic hypoxia, activation of HIF-2α may overcome the bone marrow retention signal provided by stromal-derived CXCL12, thereby enabling dissemination of multiple myeloma PCs. Here we demonstrate that HIF-2α upregulates multiple myeloma PC CXCL12 expression, decreasing migration toward CXCL12 and reducing adhesion to mesenchymal stromal cells in vitro. We also found that HIF-2α strongly induced expression of the chemokine receptor CCR1 in multiple myeloma PCs. CCR1 activation potently induces multiple myeloma PC migration toward CCL3 while abrogating the multiple myeloma PC migratory response to CXCL12. In addition, increased CCR1 expression by multiple myeloma PCs conferred poor prognosis in newly diagnosed multiple myeloma patients and was associated with an increase in circulating multiple myeloma PCs in these patients. Taken together, our results suggest a role for hypoxia-mediated CCR1 upregulation in driving the egress of multiple myeloma PCs from the bone marrow. Targeting CCR1 may represent a novel strategy to prevent dissemination and overt relapse in multiple myeloma. Cancer Res; 77(20); 5452–63. ©2017 AACR.

Type I collagen is a ubiquitous structural protein in animal tissues. It is normally present in a hydrated form. However, collagen is very dependent on associated water for its mechanical properties. In skin, where type I collagen is dominant, there is a longstanding concern that the skin and therefore collagen may partially dry out and result in structural degradation. Here we show that dehydration of type I collagen fibrils, using 2-propanol, results in a two-stage dehydration process. Initially, the fibrils do not change length, i.e. the D-period remains constant, but shrinkage occurs within the fibrils by an increase in the gap region and a decrease in the overlap region within a D-band and a shortening of the helical turn distance and fibril diameter. Only with further dehydration does the length of the collagen fibril decrease (a decrease in D-period). This mechanism explains why collagen materials are resistant to gross structural change in the early stages of dehydration and shows why they may then suffer from sudden external shrinkage with further dehydration.

Pioglitazone-loaded PLGA-PEG nanoparticles (NPs) were stabilized by the spray drying technique as an alternative to the treatment of ocular inflammatory disorders. Pioglitazone-NPs were developed and characterized physiochemically. Interaction studies, biopharmaceutical behavior, ex vivo corneal and scleral permeation, and in vivo bioavailability evaluations were conducted. Fibrillar diameter and interfibrillar corneal spacing of collagen was analyzed by synchrotron X-ray scattering techniques and stability studies at 4 °C and was carried out before and after the spray drying process. NPs showed physicochemical characteristics suitable for ocular administration. The release was sustained up to 46 h after drying; ex vivo corneal and scleral permeation profiles of pioglitazone-NPs before and after drying demonstrated higher retention and permeation through cornea than sclera. These results were correlated with an in vivo bioavailability study. Small-angle X-ray scattering (SAXS) analysis did not show a significant difference in the organization of the corneal collagen after the treatment with pioglitazone-NPs before and after the drying process, regarding the negative control. The stabilization process by Nano Spray Dryer B-90 was shown to be useful in preserving the activity of pioglitazone inside the NPs, maintaining their physicochemical characteristics, in vivo bioavailability, and non-damage to corneal collagen function after SAXS analysis was observed.

Abstract Introduction: Paclitaxel, a cytotoxic agent that destroys cancer cells by stabilizing the microtubule, inhibits tube formation by human umbilical vein endothelial cells (HUVEC) at subtoxic concentrations (≤10nM). The clear separation of the antiangiogenic effect of paclitaxel from its cytotoxicity suggests the possibility of alternative mechanisms of action. Understanding the mechanisms will help materialize the combination use of antiangiogenic cytotoxic agents with targeted therapeutic agents to obtain a synergistic antiangiogenic effect in cancer treatment. Our previous findings indicated that low dose paclitaxel can inhibit RhoA activation in HUVEC and this is strongly correlated to the antiangiogenic effect of paclitaxel. We therefore postulate that RhoA, a member of the Rho GTPases family which regulates cytoskeleton reorganization and branching of blood vessels, may be the critical link underlying the antiangiogenic effect of low dose paclitaxel. Aims: The present study aims to assess the causal effect of RhoA inactivation against the antiangiogenic effect of low dose paclitaxel through molecular and pharmacological approaches. Methods: Plasmid DNA expressing fusion proteins of enhanced green fluorescent protein (EGFP) with either wild-type RhoA (WT-RhoA) or its constitutively active mutant (V14-RhoA) were constructed in-house and confirmed by sequencing. Protein expression was evaluated using fluorescent microscopy, flow cytometry and Western Blot analysis. Transfection of HUVEC was facilitated by the Amaxa's Nucleofector Technology (Lonza). The antiangiogenic effect of paclitaxel (0.1–100nM) was examined through the 24 h inhibition of tube formation by HUVEC transfected with our plasmid constructs. The pharmacological study involved the simultaneous use of 5μM 1-oleoyllysophosphatidic acid (LPA), a RhoA activator, with low dose paclitaxel in the 24 h tube formation assay. Results: Transfection of HUVEC with WT-RhoA and V14-RhoA yielded EGFP fluorescence in 52 and 68% of the HUVEC, respectively. The antiangiogenic effect of 5, 10 and 100nM paclitaxel against HUVEC expressing V14-RhoA was reduced by 7, 56 and 6%, respectively when compared to WT-RhoA. Failure in RhoA inactivation led to modest attenuation of the antiangiogenic effect of paclitaxel. Subsequent investigation using LPA showed significant reversal of the antiangiogenic effect of low dose paclitaxel. Tube inhibition in the presence of LPA was attenuated by 89 and 42% for HUVEC treated with 0.1 and 10nM paclitaxel, respectively. Conclusions: The antiangiogenic effect of low dose paclitaxel is dependent, to a great extent, upon RhoA inactivation. We propose in-depth studies exploring the possible effect of the interaction of RhoA and other related proteins against the therapeutic effect of low dose paclitaxel. The antiangiogenic effect of paclitaxel may also be enhanced by combination with targeted therapy that inhibits RhoA activation. Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the AACR-NCI-EORTC International Conference: Molecular Targets and Cancer Therapeutics; 2011 Nov 12-16; San Francisco, CA. Philadelphia (PA): AACR; Mol Cancer Ther 2011;10(11 Suppl):Abstract nr A4.

Abstract Background: Allogeneic hematopoietic cell transplantation (allo-HCT) remains the only potentially curative therapy for high-risk hematologic malignancies. Over the last three decades, transplant numbers have increased due to expanded age eligibility, the availability of alternative donors, adoption of reduced-intensity conditioning (RIC), and improved supportive care. Despite these advances, relapse remains the major barrier to cure, and post-relapse survival is poor. Salvage strategies include donor lymphocyte infusion (DLI) or a second allo-HCT (allo-HCT2), yet no randomized trial has compared these approaches. We evaluated temporal trends in allo-HCT outcomes and survival following relapse in a State-wide cohort. Methods: We retrospectively analyzed 864 consecutive allo-HCTs performed in South Australia from 1990 to 2023. Outcomes included overall survival (OS), non-relapse mortality, and relapse, assessed using Kaplan–Meier, competing risk, and multivariable Cox regression. Post-relapse outcomes were compared for patients receiving DLI versus allo-HCT2. Results: Allo-HCT practices evolved substantially over 35 years. Median patient age increased from 41 to 54 years, with patients ≥60 years rising from &amp;lt;15% to 41% of transplants in the most recent decade. Alternative donor use increased to &amp;gt;25%, and RIC was used in &amp;gt;70% of recent transplants. Three-year OS improved significantly from 40.5% (1990–1999) to 58.0% (2020–2023; P=0.0005), primarily due to a marked reduction in non-relapse mortality from 55.3% to 24.6% (P&amp;lt;0.0001). This was validated on a multivariable analysis, transplantation in the most recent decade independently predicted superior OS (HR 0.50, P&amp;lt;0.01). In contrast, relapse risk remained ~30% since the early 2000s, highlighting it as the key barrier to cure. Among 96 patients relapsing after first allo-HCT, 40 received DLI, 43 underwent second allo-HCT2, and 13 received DLI and allo-HCT2. Median OS was 15.5 months with DLI versus 8.6 months with allo-HCT2 (P=0.59). By disease subtype, OS was similarly poor for AML, MDS, and lymphoid malignancies regardless of salvage strategy. In contrast, myeloproliferative neoplasm (MPN) patients experienced longer OS with DLI compared to allo-HCT2 (167.5 vs 7.6 months; P=0.10), although not statistically significant probably due to small number of patients. High donor CD13+ chimerism (&amp;gt;90%) prior to DLI predicted superior survival, whereas no such association was seen with allo-HCT2. Conclusion: Allo-HCT survival has improved significantly over 35 years, largely due to reduced non-relapse mortality, despite increasing patient age and complexity. Relapse remains the principal limitation, and post-relapse outcomes are poor. DLI may offer selective benefit in MPN and in myeloid patients with high donor chimerism, supporting its evaluation and incorporation into risk-adapted post-relapse algorithms.

The exponential growth of artificial intelligence (AI) in the digital world has urged researchers to use it for the betterment of society. The sensing world has also integrated AI with various low-cost, non-invasive sensors to optimise performance for real-time decision-making, thereby improving response accuracy and enhancing the scalability and adaptability of the developed AI models. This paper highlights the integration of various machine learning (ML) algorithms with sensors for food industry products. It demonstrates the mechanisms, capabilities, and advantages of each algorithm with significant examples, tailored to the target application. A classification has been performed using ML techniques commonly associated with food-related sensors. Furthermore, the capability of each of these algorithms to successfully operate on sensors with different sensing mechanisms has also been demonstrated. The influence of AI on the food industry has been demonstrated through specific examples that highlight improved food quality and safety. Finally, the paper also highlights the challenges faced by current AI-based food sensors and possible remedies. • The paper presents a topical review of the use of AI-based models in the food industry. • The use of certain machine learning (ML) models has been demonstrated in relation to the sensing solution in the food industry. • The performance of these models with respect to the food products has been shown. • The emerging trends and considerations in the AI world have been highlighted. • Solutions have been proposed to enhance the utilisation of AI-integrated sensors in the food industry.

Abstract Epigenetic reprogramming, particularly aberrant DNA methylation, drives many myeloid malignancies. While some patients have identifiable methylation modifier mutations (e.g. DNMT3a, TET2, ASXL1, or IDH1/2), many develop malignancies without such mutations. Disease progression is marked by widespread methylation changes that disrupt hematopoietic differentiation, yet the specific regulatory elements affected remain poorly defined. Furthermore, studies focusing on promoter CpG island (CGI) methylation have shown weak correlations with gene expression. This has hindered development of robust, clinically relevant methylation-based biomarkers associated with regulatory elements. Linking DNA methylation to function requires identifying functional regulatory elements. We recently reported Methylation Mesa (MM; preprint PMID: 39483908), a novel class of narrow-width (∼45–300 bp) regulatory elements with conserved methylation signatures in human and murine genomes. MM function as epigenetic switches for gene activation independent of CpG Islands. Focal core demethylation at a MM induces local histone modifications and chromatin reorganization, establishing new interaction loops inducing gene expression. Precise CRISPR-based demethylation, using CRISPR-DiR, mediated targeted demethylation reactivating silenced genes such as the tumor suppressor P16, confirming a regulatory role. To investigate epigenetic progression in Chronic Myelomonocytic Leukemia (CMML), we profiled MM in 27 adults from the PREACH clinical trial (ACTRN12621000223831), a phase 2/3 non-randomized, open-label study of newly diagnosed CMML patients stratified by mutation. Patients with RAS-pathway mutations (NRAS, KRAS, CBL) received 24 cycles of Azacitidine (Aza; 75 mg/m² SC, d1–7) and Lenzilumab (Lenz; 552 mg IV; d1 &amp; d15 of cycle 1, then d1 per cycle). Matched whole-genome bisulfite sequencing (~33x coverage) and RNA-Seq (~78M paired reads) were performed at diagnosis and after 4 treatment cycles. Our analysis focused on Mesa with a methylation delta of ≥20% compared to flanking regions. Our findings suggest that the number of Mesa triggered post-treatment can stratify patient response. Responders exhibited significantly more triggered Mesa than non-responders (mean 314,000 vs 141,000; p = 0.01, Mann-Whitney). Mesa with ≥50% positional conservation within groups revealed distinct responder/non-responder signatures. Pathway enrichment identified differential regulation in hematopoiesis (CSF1/3, IL3), TP53 (BID, TP73), Warburg signaling (FOSL1, miR-23), and PTEN signaling (TP63, miR-26). Investigations utilizing our Epigenetic Engine (EEngine), which integrates 592 curated epigenetic datasets, will refine the identification of Mesas within functional grammar contexts. These refined Mesa will define a biomarker signature that predicts CMML progression by how patients will respond to treatment, setting the stage for functional validation in ex/in vivo studies. Alternatively, preliminary research in Acute Myeloid Leukemia (AML) using a THP1 DNMT3a-R882 dox-inducible model has revealed 3 key observations regarding Mesa dynamics following R882 induction: 1) Mesa Methylation Regulation: R882mut results in Mesa loci failing to maintain basal methylation despite stable flanking methylation, implicating DNMT3amut in Mesa-specific gene regulation. 2) Metabolic Rewiring: After 3 days Aza treatment, DNMT3amut samples exhibited distinct biosynthetic and salvage pathway rewiring. 3) Aberrant Growth and Cell Cycling: Signaling and metabolic circuits promoting aberrant proliferation were activated by day 3 and sustained through day 10 of treatment. EEngine analysis highlighted PAK and heme signaling components for prioritized ex and in vivo validation. In summary, we highlight the potential of Mesa signatures to identify novel functional epigenetic elements in myeloid and other malignancies. In CMML, distinct Mesa signatures correlate with patient response to treatment. These signatures could pinpoint epigenetic events necessary for effective therapy. Similarly, in MDS and AML, identification of Mesa elements could identify functional DNA methylation sensitive elements accounting for induction of disease as well as a potential therapeutic approach. Such research aims to uncover functional elements and epigenetic biomarker signatures that are directly linked to disease stage, progression, and responsiveness to treatment

Messenger RNA lipid nanoparticles (mRNA-LNPs) have achieved remarkable success in clinical vaccination efforts to curb the COVID-19 pandemic, and have attracted tremendous interest from both industry and academia to broaden their biomedical applications. However, their storage and transportation rely heavily on the cold chain to prevent rapid degradation in non-frozen solutions, posing significant challenges for logistics and accessibility. Therefore, enhancing the in-solution stability of mRNA LNPs is crucial. Herein, the role of helper phospholipids is investigated in storage stability of mRNA LNPs. By ratiometrically replacing the helper phospholipids (DSPC) with other alternative lipids (DOPS, DOTAP, and DOPE) in commercially available mRNA LNP formulation, enhanced in-solution stability was observed with cationic lipid (DOTAP) substitution at 42 °C, room temperature (22 °C), and 4 °C. Further correlation of stability with microstructure analysis using cryogenic electron microscopy revealed that partial replacement with DOTAP enhances structural stability by promoting the formation of intact LNPs and reducing the occurrence of "bleb"-like structures. Overall, the study provides insights into the structure-stability relationship in mRNA LNPs, and offers new strategies to address the storage limitations of mRNA-LNP products in the future.

Abstract Background: TP53-mutated myeloid neoplasms (MNs) represent one of the most lethal subgroups of blood cancers. The World Health Organization (WHO-5) recognize TP53-mutated (TP53mut) myelodysplastic syndrome (MDS) and International Consensus Classification (ICC) recognize TP53mut MDS and acute myeloid leukaemia (AML) as distinct disease entities, acknowledging their dismal prognosis and the urgent need for novel therapeutic strategies. However, key discrepancies exist between the classifications, creating uncertainty in diagnosis, influencing therapeutic decisions, and complicating clinical trial enrolment for patients with uniformly poor survival. Importantly, neither classifications included TP53-mutated myelofibrosis (MF), other myeloproliferative neoplasms (MPN), and MDS/MPN overlap syndromes, despite growing evidence that these cases carry similar adverse outcomes. To address these critical gaps, we conducted a comprehensive analysis across the full spectrum of TP53-mutated MN. Methods: We included TP53mut cases with variant allele frequency (VAF ≥2%) of AML, MDS, MDS/MPN overlap, myelofibrosis and other MPN managed at the Mayo clinic (Rochester, USA) and Local Health Network in South Australia (Australia). Cases were classified according to the revised 4th edition WHO classification (WHO-4R). Biallelic TP53 loss was defined as: (i) ≥2 mutations with VAF ≥2%; (ii) single TP53mut with VAF ≥2% plus 17p loss on karyotype, FISH, or SNP array; or (iii) single TP53mut with VAF ≥50% with or without 17p loss. Monoallelic loss was defined as a single TP53mut with VAF &amp;lt;50% without 17p loss or copy neutral loss of heterozygosity. The primary endpoint was overall survival (OS) from diagnosis, censored at allogeneic transplantation. Conditional inference tree analysis was performed to define the effect of individual predictors on OS. Results: We identified 853 TP53mutMN cases including WHO-4R-defined AML (BM/PB blasts ≥20%, n=314; 36.8%), MDS with excess blasts-2 (EB-2; BM 10–19% or PB 5–19%, n=129; 15.1%), MDS-EB-1 (BM 5–9% or PB 2–4%, n=100; 11.7%), and MDS with low blasts (LB; BM &amp;lt;5% and PB &amp;lt;1%, n=228; 26.7%). MF included chronic phase (MF-CP, n=48; 5.6%), accelerated phase (MF-AP, n=7; 0.8%), and blast phase (MF-BP, n=17; 2.0%). Additional cases comprised other MPN-chronic phase (n=3, 0.4%) and MDS/MPN overlap syndromes (n=7, 0.8%). Of 848 evaluable TP53mut MN, 616 (72.6%) harbored biallelic TP53 loss and 232 (27.4%) had monoallelic loss, with 42.4% (n=98) of monoallelic cases harboring a complex karyotype (CK). Biallelic loss was highly prevalent in AML (83.1%), MDS (70.4%), and MDS/MPN overlap (71.4%), but was less frequent in cases with MPN (33.3%) and MF-chronic phase (25.0%). Across the spectrum, the frequency of biallelic loss increased stepwise with disease progression, rising from MDS-LB (62.8%) to MDS-EB1 (76.0%), MDS-EB2 (79.5%), AML (83.1%), and MF-AP/BP (79.2%) (P&amp;lt;0.0001). In multivariable analysis, WHO-4R category, TP53mut VAF, biallelic loss and CK were independent predictors of poor survival in TP53mut MN. Conditional inference tree analysis stratified the cohort into seven prognostic groups with distinct median OS: (1) AML, MF-BP/AP, and MDS/MPN overlap regardless of mono/biallelic and CK status (n=335, 39.7%; median OS 4.0 months); (2) MDS-EB1/EB2 with biallelic loss or monoallelic plus CK (n=212, 25.1%; 7.2 months); (3) MDS-LB, MF-CP and MPN with CK (n=156, 18.5%, OS 10.2 months); (4) biallelic MDS-LB (n=28, 3.3%; OS 15.6 months), (5) monoallelic MDS-EB1/EB2 without CK (n=20, 2.4%; OS 26.2 months), (6) monoallelic MDS-LB without CK (n=57, 6.7%; OS 33.4 months), and (7) MF-CP and other MPN without CK (n=36, 4.3%; OS 37.0 months). This analysis defines a clear stepwise risk gradient, identifying subsets with extremely poor prognosis versus a relatively indolent disease. Conclusions: This is the first study to evaluate the prognostic impact of TP53mut MN across the entire MN spectrum, providing a framework for future refinement of TP53-driven disease classification. Key findings include: (i) MF-AP/BP and MDS/MPN overlap exhibit survival outcomes as dismal as AML, irrespective of the TP53 allele status; and (ii) complex karyotype is a surrogate for “biallelic equivalent”, refining risk interpretation in monoallelic cases. Overall, our study defines seven prognostic subgroups spanning all TP53mut MN, offering a foundation for risk-adapted clinical trial design and future classification updates.

Reproduction, Fertility and Development is an international journal publishing original research , review and comment in the fields of reproduction and developmental biology in humans, domestic animals and wildlife