The testicular functions of neuropeptide Y are largely unexplored in non-mammalian vertebrates as the few reports available remain restricted to fishes. Current research work reveals the testicular presence of npy and npyr in Hemidactylus flaviviridis and its variation in expression depending upon reproductive phases, marking the first documentation of such findings in reptilian species. Expression analysis of npy/npyr across different reproductive phases showed peak levels of both ligand and receptor during the recrudescent period, while the lowest expression occurred in regression. The investigation also examined the influence of NPY on markers of testicular spermatogenesis and steroidogenesis in H. flaviviridis. The in vitro exposure of wall lizard testes to NPY during recrudescent phase stimulated bcl 2, while simultaneously reduced caspase 3. NPY treatment also stimulated testicular expression of scf, c-kit, pcna, bmp-15 and gdf-9, along with gonadotropin receptor (fshr), and sex-steroid receptors (er-α, er-β, and ar). Further, NPY elevated star expression and promoted testosterone synthesis in wall lizard. Regarding the regulation of testicular npy and its receptor, npyr the expression of both ligand and its receptor was upregulated by the neuropeptide, kisspeptin, adipokines like leptin and nesfatin-1, and sex steroids such as DHT, and E2, while they were suppressed by the gonadotropin, FSH and the neuropeptide, substance P. Overall, this investigation provides a detailed documentation of the presence, phase-specific pattern of expression, role, and hormonal regulation of NPY and its receptor in wall lizard testes.

Background/Objectives: Androgenetic alopecia (AGA) is the most prevalent form of hair loss. Although topical minoxidil (MNX) is widely used to treat AGA, its efficacy is limited by poor penetration across the stratum corneum. This study evaluated radiofrequency (RF) microporation as a means of enhancing cutaneous MNX delivery and hair-regrowth efficacy in a dihydrotestosterone (DHT)-induced AGA mouse model. Methods: RF-induced skin permeabilization and barrier recovery were assessed in rats using methylene blue and rhodamine B staining. In vivo skin deposition and pharmacokinetic studies were conducted to quantify cutaneous MNX accumulation and systemic exposure. Hair-regrowth efficacy was evaluated in DHT-treated mice. Results: RF microporation generated transient microchannels in the stratum corneum, increased rhodamine B penetration into deeper skin layers, and allowed substantial barrier recovery within 24 h. RF pretreatment significantly increased MNX deposition in the epidermis/dermis by 2.40-fold at 1 h and 1.94-fold at 3 h compared with topical MNX alone. RF-assisted topical administration resulted in a relative bioavailability of 11.17%, compared with 4.74% for topical administration without RF, while dose-normalized systemic exposure remained substantially lower than that following oral administration. In the AGA model, RF-assisted MNX treatment significantly increased hair coverage, length, and shaft thickness. Histological analysis further showed more prominent follicular structures in the RF-assisted MNX groups. Conclusions: RF microporation creates transient microchannels that enhance cutaneous MNX delivery and improve hair-regrowth efficacy. Importantly, RF-assisted topical administration maintained substantially lower systemic exposure than oral administration, supporting its potential as a needle-free strategy for topical AGA therapy and further translational evaluation.

Background: Sex hormones play crucial functions in the body via the genomic and non-genomic pathways. 5α- and 5β-dihydrotestosterone (5α- and 5β-DHT) are 5-reduced testosterone metabolites. We aimed to investigate the non-genomic effect of 5α- and 5β-DHT on uterine muscle contractility in vitro and in vivo for non-pregnant and 22-day-pregnant rats. Methods: The rapid in vitro action of 5α-DHT and 5β-DHT (10[-9]-10[-3] M) on KCl (25 mM)-stimulated contractions was examined in an organ bath in the presence of several blockers and after endometrium removal. The actions of DHTs (10[-4] M) and nifedipine (10[-7] M) were also examined in contractions stimulated by KCl (40 mM) with a cumulative addition of CaCl2 (3-120 mM). Plasma DHT levels were measured by ELISA after a single intraperitoneal (i.p.) administration of DHT (10 mg/kg), and kinetic curves were obtained. The in vivo relaxing action of DHTs was detected by strain-gauge sensors. The animals received 5α- or 5β-DHT alone (3/10/30/100/300 mg/kg i.p.) or with flutamide (100 mg/kg i.p.). Results: DHT showed concentration-dependent relaxation of uterine muscle in vitro, with a higher potency observed for 5β-DHT. Among the blockers used, G15 and L-NAME reduced the potency of 5α-DHT in pregnant rats only. Both DHTs inhibited the contraction-increasing effect of CaCl2, proving their Ca[2+] inhibiting effects. DHTs had similar cmax and tmax values in both non-pregnant and pregnant rats. DHT plasma levels before and 30 min after administration were proportional to the administered doses. Their single doses (30/100/300 mg/kg) elicited a flutamide-resistant uterine relaxing effect in vivo. Conclusions: DHTs or their analogs are candidates for further studies in the human uterus to establish their potential to treat conditions associated with uterine hyperactivity.

The development of sustainable, biocompatible scaffolds with enhanced structural stability remains a primary challenge in bone tissue engineering. In this study, structurally reinforced nanocomposite scaffolds were successfully fabricated by integrating green-synthesized carbon quantum dots (CQDs) into a gelatin (G) matrix, offering an innovative platform that mimics the organic-inorganic interfaces of natural bone tissue. The CQDs were derived from oatmeal via a sustainable, green hydrothermal route, serving simultaneously as zero-dimensional reinforcing fillers and bioactive agents within the biopolymer network. To ensure an additive-free fabrication process that avoids toxic chemical cross-linkers, dehydrothermal (DHT) treatment was employed, successfully modulating the interfacial and chemical cross-linking interactions between the gelatin chains and the oxygen-rich surface groups of the CQDs. Structural characterization confirmed the uniform dispersion of CQDs (average diameter 7-8 nm) within the porous gelatin framework. The incorporation of CQDs significantly improved the physicochemical properties of the scaffolds; the G/CQD 5% formulation emerged as the optimal composition, exhibiting a 118% increase in compression modulus compared to pristine gelatin. The composite demonstrated tuned swelling kinetics and a significantly reduced degradation rate, restricting mass loss after 14 days of incubation to approximately 24% compared to 40% for pristine gelatin, which is essential for maintaining a structural template during the initial stages of tissue formation. Bioactivity assays in simulated body fluid (SBF) confirmed the rapid, biomimetic induction of a crystalline hydroxyapatite layer with a natural Ca/P ratio of 1.61 within 14 days. Furthermore, preliminary in vitro assessments using Human Skin Fibroblasts (HSFs) confirmed excellent general cytocompatibility, with cell viability exceeding 90%. This study highlights the unique potential of utilizing biomass-derived carbon nanostructures and clean manufacturing processing to engineer multifunctional scaffolds with enhanced structural stability and intrinsic bioactivity for potential bone defect repairs.

Autism spectrum disorder (ASD) exhibits a profound male biased sex ratio. While numerous genes have been implicated in ASD, the functional basis of this sex difference is unclear. One enticing hypothesis is genome-wide transcriptional regulation through estrogens and androgens. While hormone-mediated transcription is well-studied in reproductive tissues, its role in cortical development is poorly defined. Thus, we profiled androgen (AR) and estrogen (ESR1/ESR2) receptor expression in primary human cortex and developmental stage-matched organoid models by single-cell RNA-seq. AR was primarily expressed in radial glia and intermediate progenitors while ESR1/ESR2 was more broadly distributed across multiple cell types of the developing cortex, although with the highest expression in radial glia. To study their genetic effects, we exposed iNeurons and cortical organoids to physiological levels of dihydro-testosterone (DHT) and estradiol (E2). DHT consistently up-regulated oxidative metabolism programs enriched in progenitor cells and down-regulated neuronal maturation pathways, while E2 exhibited a much more attenuated effect. The presence of DHT reduced NTRK2 (TrkB) expression, correlating with expression in developing cortex where NTRK2 had significantly higher expression in progenitor cells of the female cortex, which is also reflected in the increased expression of AR in radial glia. Together, these data indicate that in developing human cortical lineages, sex hormones act as selective, cell-state-dependent modulators that tune metabolic and maturation programs rather than broadly reprogramming the genome. Thus, the effects of variation in transcriptional regulation through estrogens and androgens are likely to be minor, but not absent, in ASD.

Hair follicles, as key accessory organs in mammalian skin, are precisely regulated for growth and development influenced by genetic factors, hormones, and the local microenvironment. This study investigated the effect of dihydrotestosterone (DHT) on the proliferation of dermal papilla cells (DPCs) from Xinjiang Hetian sheep and its regulatory relationship with the TGF-β/Smad signalling pathway. DPCs were isolated from the back skin tissue of newborn Hetian sheep by collagenase II digestion, and their characteristics were identified by primary culture, passage, and double immunofluorescence staining with α-SMA and CD133 labelling. CCK-8 and EdU assays were used to evaluate the effect of varying DHT concentrations (0, 1, 10, 100, and 1000 nM) on DPC proliferation. The results showed that with the increased DHT concentration, cell proliferation also increased, and the highest rate was observed at 1000 nM. Furthermore, qPCR analysis revealed that 1000 nM DHT significantly upregulated Smad6 gene expression (p < 0.05), while it inhibited the expression of TGF-β1, TGF-βRII, and Smad5 (p < 0.05). These findings suggest that high DHT concentrations may promote DPC proliferation by inhibiting the TGF-β/Smad signalling pathway. This suggests a critical role of this pathway in regulating hair follicle development and hormone-dependent hair growth, providing evidence for the molecular mechanism of androgen action on dermal papilla cell function.

Androgenetic alopecia (AGA) is the most common form of hair loss, with a high prevalence worldwide. In this study, we aimed to investigate the potential of Tenebrio molitor larva trypsin hydrolysate (TMTH), an insect-derived bioactive hydrolysate, as a functional ingredient candidate for AGA. In vitro and in vivo assessments demonstrated that TMTH promoted human hair dermal papilla (HDP) cell proliferation, restored β-catenin expression, and alleviated dihydrotestosterone (DHT)-induced inhibition. In a testosterone propionate (TP)-induced alopecia mouse model, oral TMTH administration increased hair coverage and follicle number, upregulated Wnt/β-catenin and PI3K/Akt signaling, reduced androgen receptor expression, enhanced antioxidant-related responses, and suppressed inflammatory markers. These findings suggest that TMTH may ameliorate AGA-related phenotypes by modulating hair growth-related signaling, local androgen-related responses, oxidative stress, and inflammation, thereby supporting its potential as a functional ingredient candidate for hair loss management.

This study investigated the effects of dry-heat treatment (DHT) temperature on whole quinoa flour structure, dough rheology, bread texture, starch digestibility, and hypoglycemic potential in STZ-induced diabetic mice. DHT induced surface aggregate detachment and imperfections on flour particles, while preserving the A-type starch crystallinity. Compared to untreated controls, DHT (110-150 °C) increased dough water absorption and softening degree, but decreased extensibility and resistance. Bread made with 110 °C-treated flour showed a slight decrease in hardness and a significant improvement in springiness; higher temperatures progressively deteriorated the texture. DHT notably lowered rapidly digestible starch and elevated slowly digestible and resistant starch fractions. Based on these in vitro results, 110 °C was identified as the optimal temperature, and only the 110 °C-treated flour was used for in vivo evaluation. In STZ-induced diabetic mice, six-week quinoa bread supplementation suggested beneficial effects on glycated hemoglobin, blood glucose, and insulin levels. These findings provide a theoretical basis for designing quinoa-based functional foods.

Azole fungicides are utilized in global agriculture to protect rice, vegetables, and fruits from fungal pathogens, yet their capacity to interfere with neurosteroid biosynthesis has raised significant toxicological concerns. In this study, we investigated the effects of eleven representative azole fungicides on the enzymatic activity of 5α-reductase 1 (SRD5A1) using both human and rat models. To assess inhibition, we employed HPLC-MS/MS to precisely measure testosterone to dihydrotestosterone conversion, complemented by SPR, intact cell assays, in silico docking, molecular dynamics, and 3D-QSAR modeling. Our screening revealed that three azoles inhibited the human isoform, whereas four inhibited the rat counterpart, with ipconazole demonstrating the strongest potency (IC50 = 3.40 μM for human, 15.24 μM for rat). SPR confirmed its tight binding (KD = 2.12 μM) and NADPH disruption. Kinetic analyses classified these as mixed/noncompetitive inhibitors. In SF126 cells, 10 and 100 μM exposures markedly reduced DHT output. SAR correlated efficacy with LogP, and 3D-QSAR highlighted hydrophobic interactions as critical. Docking revealed that azoles occupy the NADPH-binding site via van der Waals forces, hydrogen bonds, hydrophobic contacts, and charge interactions. Collectively, our findings demonstrate that azole fungicides inhibit SRD5A1 across species, potentially causing neural disorders by depleting neurosteroids, underscoring the need for further neurotoxicological research.

This study investigates how pre-esterification dry heat treatment (DHT) of corn starch at 170°C, 180°C, and 200°C influences the functional properties of subsequently synthesized starch citrates. Structural and morphological changes induced by DHT and esterification were analyzed using scanning electron microscopy, X-ray diffractometry, and Fourier-Transform Infrared spectroscopy. Starch citrates were characterized for their resistance to pancreatic α-amylase and amyloglucosidase, as well as their water absorption capacity (WAC) at 37°C. Results demonstrated a strong correlation between DHT temperature and the functional properties of the final citrate. Increasing the DHT temperature significantly increased the resistant starch fraction and WAC. Specifically, citrates from native starch had a resistant starch (RS) fraction of 91%, while citrates from starch pre-treated at 200°C exhibited a markedly increased RS fraction of 98% and a 64% increase in WAC. These findings indicate that DHT is an effective pre-treatment for enhancing the dietary fiber potential and hydration properties of starch citrates.

Periodontal tissue regeneration requires coordinated cellular responses among multiple cell types, supporting matrix components, and signaling proteins, including bone morphogenetic proteins (BMPs). Despite extensive studies on BMP2, the regenerative role of BMP9 and its interplay with BMP2 in shaping bone and cementum repair remain poorly understood. Notably, BMP9 was independent of the antagonist Noggin, and molecular dynamics simulations confirmed that the Noggin-BMP9 complex exhibited greater root mean square deviation (RMSD) values and unstable interactions in periodontal ligament stem cells (PDLSCs). Functionally, BMP9 notably enhanced the osteogenic and cementoblastic differentiation of PDLSCs, elicited minimal osteoclast activation, and improved the survival of PDLSCs and macrophages by attenuating stress-induced apoptosis and preserving mitochondrial membrane potential. BMP9 also induced epithelial-mesenchymal transition (EMT) and exhibited synergy with endogenous matrix proteins to enhance mineralization. To optimize BMP localization in the application site, sponge type collagen scaffolds were attempted. Type I collagen sponges crosslinked with carbodiimide (EDC/NHS) and dehydrothermal (DHT) were compared, and the DHT-crosslinked sponge was selected for its larger pores, higher absorbency, and higher BMP9 loading efficiency. In a canine periodontal defect model, BMP9-loaded DHT-collagen sponges facilitated integrated regeneration of cementum, periodontal ligament, and alveolar bone, accompanied by localized ApoBD-like structures associated with PDLSC- and macrophage-related markers. Transcriptomic profiling revealed notable induction of gene sets related to EMT and cementogenesis, underscoring the role of BMP9 in driving periodontal tissue homeostasis. Collectively, these findings establish high-affinity BMP9 delivery via a collagen sponge as an effective and clinically translatable strategy for periodontal tissue regeneration.