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Molecules (Basel, Switzerland) Mar 2020Novel reduction-responsive hyaluronic acid-chitosan-lipoic acid nanoparticles (HACSLA-NPs) were designed and synthesized for effective treatment of breast cancer by...
Novel reduction-responsive hyaluronic acid-chitosan-lipoic acid nanoparticles (HACSLA-NPs) were designed and synthesized for effective treatment of breast cancer by targeting Cluster of Differentiation 44 (CD44)-overexpressing cells and reduction-triggered 17α-Methyltestosterone (MT) release for systemic delivery. The effectiveness of these nanoparticles was investigated by different assays, including release rate, 3-(4,5-Dimethylthiazol-2-Yl)-2,5-Diphenyltetrazolium Bromide (MTT), lactate dehydrogenase (LDH), caspase-3 activity, Rhodamine 123 (RH-123), and Terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL). In vitro experiments revealed that Methyltestosterone/Hyaluronic acid-chitosan-lipoic acid nanoparticles (MT/HACSLA-NPs) illustrated a sustained drug release in the absence of glutathione (GSH), while the presence of GSH led to fast MT release. HACSLA-NPs also showed high cellular internalization via CD44 receptors, quick drug release inside the cells, and amended cytotoxicity against positive CD44 BT-20 breast cancer cell line as opposed to negative CD44, Michigan Cancer Foundation-7 (MCF-7) cell line. These findings supported that these novel reduction-responsive NPs can be promising candidates for efficient targeted delivery of therapeutics in cancer therapy.
Topics: Antineoplastic Agents, Hormonal; Biomarkers; Cell Line, Tumor; Drug Carriers; Drug Delivery Systems; Female; Humans; Hyaluronic Acid; Hydrogen-Ion Concentration; Magnetic Resonance Spectroscopy; Methyltestosterone; Nanoparticles; Oxidation-Reduction; Spectroscopy, Fourier Transform Infrared; X-Ray Diffraction
PubMed: 32151062
DOI: 10.3390/molecules25051181 -
PloS One 2020Fish oogenesis is characterised by a massive growth of oocytes each reproductive season. This growth requires the stockpiling of certain molecules, such as ribosomal...
Fish oogenesis is characterised by a massive growth of oocytes each reproductive season. This growth requires the stockpiling of certain molecules, such as ribosomal RNAs to assist the rapid ribosomal assembly and protein synthesis required to allow developmental processes in the newly formed embryo. Massive 5S rRNA expression in oocytes, facilitated by transcription factor 3A (Gtf3a), serves as marker of intersex condition in fish exposed to xenoestrogens. Our present work on Gtf3a gene evolution has been analysed in silico in teleost genomes and functionally in the case of the zebrafish Danio rerio. Synteny-analysis of fish genomes has allowed the identification of two gtf3a paralog genes, probably emerged from the teleost specific genome duplication event. Functional analyses demonstrated that gtf3ab has evolved as a gene specially transcribed in oocytes as observed in Danio rerio, and also in Oreochromis niloticus. Instead, gtf3aa was observed to be ubiquitously expressed. In addition, in zebrafish embryos gtf3aa transcription began with the activation of the zygotic genome (~8 hpf), while gtf3ab transcription began only at the onset of oogenesis. Under exposure to 100 ng/L 17β-estradiol, fully feminised 61 dpf zebrafish showed transcription of ovarian gtf3ab, while masculinised (100 ng/L 17α-methyltestosterone treated) zebrafish only transcribed gtf3aa. Sex related transcription of gtf3ab coincided with that of cyp19a1a being opposite to that of amh and dmrt1. Such sex dimorphic pattern of gtf3ab transcription was not observed earlier in larvae that had not yet shown any signs of gonad formation after 26 days of oestradiol exposure. Thus, gtf3ab transcription is a consequence of oocyte differentiation and not a direct result of estrogen exposure, and could constitute a useful marker of gonad feminisation and intersex condition.
Topics: Animals; Cichlids; Disorders of Sex Development; Evolution, Molecular; Female; Fish Proteins; Gene Duplication; Male; Oogenesis; Ovary; Phylogeny; Sex Characteristics; Sex Differentiation; Synteny; Transcription Factor TFIIIA; Zebrafish; Zebrafish Proteins
PubMed: 31999691
DOI: 10.1371/journal.pone.0227690 -
BMC Genomics Jan 2020Spermatogenesis is an intricate process regulated by a finely organized network. The orange-spotted grouper (Epinephelus coioides) is a protogynous hermaphroditic fish,...
Transcriptome profiling of laser-captured germ cells and functional characterization of zbtb40 during 17alpha-methyltestosterone-induced spermatogenesis in orange-spotted grouper (Epinephelus coioides).
BACKGROUND
Spermatogenesis is an intricate process regulated by a finely organized network. The orange-spotted grouper (Epinephelus coioides) is a protogynous hermaphroditic fish, but the regulatory mechanism of its spermatogenesis is not well-understood. In the present study, transcriptome sequencing of the male germ cells isolated from orange-spotted grouper was performed to explore the molecular mechanism underlying spermatogenesis.
RESULTS
In this study, the orange-spotted grouper was induced to change sex from female to male by 17alpha-methyltestosterone (MT) implantation. During the spermatogenesis, male germ cells (spermatogonia, spermatocytes, spermatids, and spermatozoa) were isolated by laser capture microdissection. Transcriptomic analysis for the isolated cells was performed. A total of 244,984,338 clean reads were generated from four cDNA libraries. Real-time PCR results of 13 genes related to sex differentiation and hormone metabolism indicated that transcriptome data are reliable. RNA-seq data showed that the female-related genes and genes involved in hormone metabolism were highly expressed in spermatogonia and spermatozoa, suggesting that these genes participate in the spermatogenesis. Interestingly, the expression of zbtb family genes showed significantly changes in the RNA-seq data, and their expression patterns were further examined during spermatogenesis. The analysis of cellular localization of Eczbtb40 and the co-localization of Eczbtb40 and Eccyp17a1 in different gonadal stages suggested that Eczbtb40 might interact with Eccyp17a1 during spermatogenesis.
CONCLUSIONS
Our study, for the first time, investigated the transcriptome of the male germ cells from orange-spotted grouper, and identified functional genes, GO terms, and KEGG pathways involved in spermatogenesis. Furthermore, Eczbtb40 was first characterized and its role during spermatogenesis was predicted. These data will contribute to future studies on the molecular mechanism of spermatogenesis in teleosts.
Topics: Animals; Fishes; Gene Expression Profiling; Germ Cells; Methyltestosterone; Spermatogenesis; Transcriptome
PubMed: 31973692
DOI: 10.1186/s12864-020-6477-4 -
International Journal of Molecular... Aug 2019A-type K channels contribute to regulating the propagation and frequency of action potentials in smooth muscle cells (SMCs). The present study (i) identified the...
A-type K channels contribute to regulating the propagation and frequency of action potentials in smooth muscle cells (SMCs). The present study (i) identified the molecular components of A-type K channels in rat vas deferens SMs (VDSMs) and (ii) showed the long-term, genomic effects of testosterone on their expression in VDSMs. Transcripts of the A-type K channel α subunit, Kv4.3L and its regulatory β subunits, KChIP3, NCS1, and DPP6-S were predominantly expressed in rat VDSMs over the other related subtypes (Kv4.2, KChIP1, KChIP2, KChIP4, and DPP10). A-type K current (I) density in VDSM cells (VDSMCs) was decreased by castration without changes in I kinetics, and decreased I density was compensated for by an oral treatment with 17α-methyltestosterone (MET). Correspondingly, in the VDSMs of castrated rats, Kv4.3L and KChIP3 were down-regulated at both the transcript and protein expression levels. Changes in Kv4.3L and KChIP3 expression levels were compensated for by the treatment with MET. These results suggest that testosterone level changes in testosterone disorders and growth processes control the functional expression of A-type K channels in VDSMCs.
Topics: Animals; Blotting, Western; Castration; Down-Regulation; Electrophysiology; Kv Channel-Interacting Proteins; Male; Methyltestosterone; Muscle, Smooth; Rats; Rats, Wistar; Testosterone; Vas Deferens
PubMed: 31438481
DOI: 10.3390/ijms20174073 -
The Journal of Steroid Biochemistry and... Dec 2019Estrogen receptor (ER) sequences vary between species and this suggests that there are differences in the ligand-specificity, leading to species-specific effects. This...
Estrogen receptor (ER) sequences vary between species and this suggests that there are differences in the ligand-specificity, leading to species-specific effects. This would indicate that it is not possible to generalize effects across species. In this study, we investigated the differences in activation potencies and binding affinities of ER´s alpha (α) and beta (β) in human, zebrafish and sea bream to elucidate species differences in response to estradiol, estrone, estriol and methyltestosterone. In vitro analysis showed that estradiol had the highest activity for all the ER´s except for human ERβ and seabream ERβ2. Alignment of the ligand binding domain and ligand binding pocket (LBP) residues of the three species showed that different residues were involved in the LBPs which led to differences in pocket volume, affected binding affinity and orientation of the ligands. By combining in silico and in vitro results, it was possible to identify the ligand specificities of ER´s. The results demonstrated that the human ER´s show lower resolution in ligand-dependent activation, suggesting higher promiscuity, than the zebrafish and seabream ER´s. These results show species-specificity of ER´s and suggest that species-specific differences must be taken into consideration when studying different exposure scenarios.
Topics: Androgens; Animals; Cell Line; Estradiol; Estriol; Estrogens; Estrone; Fish Proteins; Humans; Ligands; Methyltestosterone; Molecular Docking Simulation; Receptors, Estrogen; Sea Bream; Species Specificity; Zebrafish
PubMed: 31437548
DOI: 10.1016/j.jsbmb.2019.105450 -
Clinical Journal of the American... Apr 2019Many aspects of CKD management rely heavily on patient self-care, including medication and dietary adherence, self-monitoring of BP, and daily physical activity. Growing...
BACKGROUND AND OBJECTIVES
Many aspects of CKD management rely heavily on patient self-care, including medication and dietary adherence, self-monitoring of BP, and daily physical activity. Growing evidence suggests that incorporating smartphone-based applications can support self-care in CKD and chronic disease more generally.
DESIGN, SETTING, PARTICIPANTS, & MEASUREMENTS
We identified applications targeting patients with CKD by conducting a search of the US Apple App Store (iOS) and Google Play Store (Android) using the following four phrases: "kidney disease," "renal," "dialysis," and "kidney transplant." We considered the first 50 applications for each search term on each application store. We adapted a previously described framework for assessment of mobile health applications to account for kidney disease-specific content areas and evaluated applications on their types of patient engagement, quality, usability, and safety. Engagement and quality were assessed by both a patient and a nephrologist, usability was assessed by a patient, and safety was assessed by a nephrologist. Overall, two patients with CKD and three nephrologists performed the evaluations. We examined pairwise correlations between patient, nephrologist, and consumer ratings of application quality.
RESULTS
Our search strategy identified 174 unique applications on Android and 165 unique applications on iOS. After excluding applications that were not related to kidney disease, were not patient facing, or were last updated before 2014, 12 Android-only applications, 11 iOS-only applications, and five dual-platform applications remained. Patient and nephrologist application quality ratings, assessed by the net promoter score, were not correlated (=0.36; =0.06). Consumer ratings on the application stores did not correlate with patient ratings of application quality (=0.34; =0.18).
CONCLUSIONS
Only a small subset of CKD applications was highly rated by both patients and nephrologists. Patients' impressions of application quality are not directly linked to consumer application ratings or nephrologist impressions.
Topics: Attitude of Health Personnel; Humans; Mobile Applications; Nephrology; Patient Participation; Renal Insufficiency, Chronic; Self Care; Smartphone
PubMed: 30898873
DOI: 10.2215/CJN.10370818 -
International Journal of Molecular... Sep 2018In this study, we systematically investigated the process of sex reversal induced by 17-methyltestosterone (MT) feeding and MT-feeding withdrawal at the ovary...
In this study, we systematically investigated the process of sex reversal induced by 17-methyltestosterone (MT) feeding and MT-feeding withdrawal at the ovary differentiation stage in orange-spotted groupers, . Gonadal histology showed that MT feeding induced a precocious sex reversal from immature ovaries to testes, bypassing the formation of an ovarian cavity, and MT-feeding withdrawal led to an ovarian fate. In both the MT feeding and MT-feeding withdrawal phases, cytochrome P450 family 11 subfamily B () gene expression and serum 11-KT levels were not significantly changed, suggesting that the MT-treated fish did not generate endogenous steroids, even though active spermatogenesis occurred. Finally, by tracing doublesex-expressing and Mab-3-related transcription factor 1 (dmrt1)-expressing cells and TUNEL (terminal deoxynucleotidyl transferase 2-deoxyuridine, 5-triphosphate nick end labeling) assays, we found that the efferent duct formed first, and then, the germ cells and somatic cells of the testicular tissue were generated around the efferent duct during MT-feeding-induced precocious sex reversal. Collectively, our findings provide insights into the molecular and cellular mechanisms underlying sex reversal induced by exogenous hormones during sex differentiation in the protogynous orange-spotted grouper.
Topics: Animals; Bass; Cytochrome P450 Family 11; Female; Fish Proteins; Germ Cells; Male; Methyltestosterone; Sex Determination Processes; Sex Differentiation; Transcription Factors
PubMed: 30235790
DOI: 10.3390/ijms19092828 -
Zoological Research Mar 2019Genetically improved farmed tilapia (GIFT) and GIFT-derived strains account for the majority of farmed tilapia worldwide. As male tilapias grow much faster than females,...
Genetically improved farmed tilapia (GIFT) and GIFT-derived strains account for the majority of farmed tilapia worldwide. As male tilapias grow much faster than females, they are often considered more desirable in the aquacultural industry. Sex reversal of females to males using the male sex hormone 17-α-methyltestosterone (MT) is generally used to induce phenotypic males during large-scale production of all male fingerlings. However, the widespread use of large quantities of sex reversal hormone in hatcheries may pose a health risk to workers and ecological threats to surrounding environments. Breeding procedures to produce genetically all-male tilapia with limited or no use of sex hormones are therefore urgently needed. In this study, by applying marker-assisted selection (MAS) for the selection of YY supermales from a GIFT-derived strain, we identified 24 XY pseudofemale and 431 YY supermale tilapias. Further performance evaluation on the progenies of the YY supermales resulted in male rates of 94.1%, 99.5% and 99.6%, respectively, in three populations, and a daily increase in body weight of 1.4 g at 3 months (n=997). Our study established a highly effective MAS procedure in the selection of YY supermales from a GIFT-derived strain. Furthermore, the development of MAS-selected YY supermales will help reduce the utilization of hormones for controlling sex in the tilapia aquaculture.
Topics: Animals; Aquaculture; Male; Selection, Genetic; Sex Determination Processes; Sex Ratio; Tilapia; Y Chromosome
PubMed: 30213922
DOI: 10.24272/j.issn.2095-8137.2018.071 -
Microbial Cell Factories May 2018Steroid compounds are very interesting substrates for biotransformation due to their high biological activity and a high number of inactivated carbons which make...
BACKGROUND
Steroid compounds are very interesting substrates for biotransformation due to their high biological activity and a high number of inactivated carbons which make chemical modification difficult. Microbial transformation can involve reactions which are complicated and uneconomical in chemical synthesis, and searching for a new effective biocatalyst is necessary. The best known entomopathogenic species used in steroid modification is Beauveria bassiana. In this study we tested the ability of Isaria farinosa, another entomopathogenic species, to transform several steroids.
RESULTS
Twelve strains of the entomopathogenic filamentous fungus Isaria farinosa, collected in abandoned mines located in the area of the Lower Silesian Voivodeship, Poland, from insects' bodies covered by fungus, were used as a biocatalyst. All the tested strains effectively transformed dehydroepiandrosterone (DHEA). We observed 7α- and 7β-hydroxy derivatives as well as changes in the percentage composition of the emerging products. Due to the similar metabolism of DHEA in all tested strains, one of them was selected for further investigation. In the culture of the selected strain, Isaria farinosa KCh KW1.1, transformations of androstenediol, androstenedione, adrenosterone, 17α-methyltestosterone, 17β-hydroxyandrost-1,4,6-triene-3-one and progesterone were performed. All the substrates were hydroxylated with high yield and stereoselectivity. We obtained 6β-hydroxyandrost-4-ene-3,11,17-trione, 15α,17β-dihydroxy-6β,7β-epoxyandrost-1,4-diene-3-one and 6β,11α-dihydroxyprogesterone. There is no evidence of either earlier microbial transformation of 17β-hydroxyandrost-1,4,6-triene-3-one or new epoxy derivatives.
CONCLUSIONS
Isaria farinosa has a broad spectrum of highly effective steroid hydroxylases. The obtained 7-hydroxydehydroepiandrosterone has proven high biological activity and can be used in Alzheimer's disease and as a key intermediate in the synthesis of aldosterone antagonists. Transformation of progesterone leads to high yield of 6β,11α-dihydroxyprogesterone and it is worth further study.
Topics: Biotransformation; Dehydroepiandrosterone; Fungal Proteins; Progesterone; Steroids
PubMed: 29753319
DOI: 10.1186/s12934-018-0920-0 -
International Journal of Molecular... Feb 2018The transcription factor has been implicated in cartilage formation and testis determination in mammals. Here, two duplicates of were found in Japanese flounder ()...
The transcription factor has been implicated in cartilage formation and testis determination in mammals. Here, two duplicates of were found in Japanese flounder () named and , respectively. Phylogenetic and gene structure analyses revealed that and were homologous to that of teleosts and tetrapods. Quantitative real-time polymerase chain reaction (qRT-PCR) showed that both and expressed higher in testis than in ovary of adult tissues. The in situ hybridization (ISH) analysis of gonads showed that and mRNA were both detected in oocytes, Sertoli cells and spermatocytes. During sex differentiation, the expression of exhibited obvious sexual dimorphic expression from 60 days after hatch (dah) with higher expression in male preferred individuals than female preferred individuals and increased gradually from 30 to 100 dah. A similar pattern was detected in expression. After injection of androgen (17α-methyltestosterone) of different concentrations, the expression level of increased significantly, whereas did not change obviously. These results indicated that the two genes of Japanese flounder had converse functions in sex differentiation, whereas their differences in 17α-methyltestosterone administration were obvious and worthwhile for exploring evolutionary and adaptive significance. This study provided a foundation for further exploration of the roles of genes during the sex determination and differentiation, spermatogenesis and gonadal function maintenance of Japanese flounder.
Topics: Amino Acid Sequence; Animals; Cloning, Molecular; Flounder; Gene Expression; Gene Expression Regulation, Developmental; Genome; Gonads; Methyltestosterone; Organ Specificity; Organogenesis; Phylogeny; RNA, Messenger; SOX9 Transcription Factor; Sequence Analysis, DNA; Sex Differentiation; Spermatogenesis
PubMed: 29419762
DOI: 10.3390/ijms19020512