Exposure of carbon nanotubes affects testis and brain of common carp.

Carbon nanotubes production has been rapidly increasing for many potential applications, however, the environmental impact of this nanomaterial needs to be comprehended. The present work focused on unraveling the effects of single-walled carbon nanotubes (SWCNT) in the common carp, Cyprinus carpio. The physicochemical properties of SWCNT were analyzed with X-ray diffraction, Fourier transforms infra-red, UV-Vis absorption, transmission electron microscopy (TEM), and Raman spectroscopy before testing for exposure impact. The effects of SWCNT, were investigated by exposing to two doses viz., 10 and 50 µg/L, for 7 days in adult common carp, in vivo. Expression of key steroidogenic and transcription factor genes related to testis and brain were downregulated after the treatment. The concomitant decreases in serum testosterone and 11-ketotestosterone levels revealed the impact of SWCNT after exposure. Further, SWCNT exposure induced antioxidant enzymes namely glutathione-S-transferases, superoxide dismutase, and catalase in both testis and brain. Concurrently, histological and TEM analysis of testis revealed structural disarray. In addition, SWCNT treatment, in testicular and brain primary cell cultures decreased cell viability with an increase of reactive oxygen species levels, leading to a significant elevation of apoptotic cells. In line with this, low mitochondrial membrane potential and DNA damage were also observed during post SWCNT treatment. Taken together, transient exposure of SWCNT causes toxic effects and alters testicular and brain function in the common carp. Thus, the discharge of carbon nanotubes poses a greater risk to the aquatic environment warranting regulatory measures.

Lactoferrin nanoparticles coencapsulated with curcumin and tenofovir improve vaginal defense against HIV-1 infection.

Aim: We report here the development of tenofovir- and curcumin-loaded lactoferrin nanoparticles (TCNPs) as an HIV-microbicide. Materials & methods: TCNPs were subjected to various physicochemical characterization experiments, followed by in vitro and in vivo experiments to assess their efficacy. Results: TCNPs had a diameter of 74.31 ± 2.56 nm with a gross encapsulation of more than 61% for each drug. Nanoparticles were effective against HIV-1 replication, with an IC50 of 1.75 µM for curcumin and 2.8 µM for tenofovir. TCNPs provided drug release at the application site for up to 8-12 h, with minimal leakage into the systemic circulation. TCNPs showed spermicidal activity at ≥200 µM and induced minimal cytotoxicity and inflammation in the vaginal epithelium as revealed by histopathological and ELISA studies. Conclusion: We demonstrated that TCNPs could serve as a novel anti-HIV microbicidal agent in rats. [Formula: see text].

Interactive role of Wnt signaling and Zn in regulating testicular function of the common carp, Cyprinus carpio.

Wnt signaling is conserved among all species and plays a significant role in various cellular processes including reproduction. The present study identified significant involvement of wnt4a, wnt5b, and wnt8a signaling in the testicular growth of common carp,Cyprinus carpio. Predominant expression of wnt4a, wnt5b, and wnt8a was found in the gonads and Wnt4a was localized in spermatocytes and interstitial cells. Ontogeny and testicular phase-wise analysis signified the importance of wnt isofoms analyzed in this study. Specific pathway activation of Wnt signaling revealed that Wnt4a and Wnt5b act through non-canonical while Wnt8a prefers the canonical pathway. The Wnt signaling regulates several steroidogenic enzyme and testis-related genes which was confirmed by the Wnt blockade experiments. Incidentally, zinc (Zn) is an essential trace element involved in the progression of spermatogenesis in teleosts. In adult male carp, a single administration of Zn at different doses elevated the expression of Wnt and Zn transporter genes and a single-dose (30 µg/g body weight [BW]) of Zn treatment elevated steroidogenic enzyme and testis-related genes which coincided with elevated androgens. Conversely, single-dose administration of Zn chelator to the Zn administered (30 µg/g BW) fish reversed the effects emphasizing a prominent role of Zn in the testicular function perhaps through Wnt signaling. Similar effects were observed in the in vitro experiments using the Zn chelator. Bioaccumulation of Zn and histological analysis revealed the importance of Zn in progression of spermatogenesis and sperm motility. Various assays related to cell viability and proliferation exhibited the role of Zn in promoting spermatogenic cell progression. Flow cytometric analysis confirmed Zn-induced elevation of Wnt and Zn transporter genes in germ and supporting cells. Furthermore, the effects of Zn are dose-related in carp. Taken together, it seems wnt4a, wnt5b, and wnt8a play an important role in testis and exposure of Wnt inhibitor, canonical as well as non-canonical activators, and Zn confirmed that Zn regulates Wnt signaling vis-à-vis promoting spermatogenesis in the common carp.

Effects of zinc oxide nanoparticles and zinc sulfate on the testis of common carp, Cyprinus carpio.

The present study analyzed the effects of zinc oxide nanoparticles (ZnO-NPs) and zinc sulfate (ZnSO4) in the testis of six-month-old common carp Cyprinus carpio exposed to three different doses, viz., 10, 50, and 100 µg/L for 21 days. Characterization of ZnO-NPs was done after sonication, the size and shape of ZnO-NPs were determined as ∼20-30 nm spherical structure measured zeta potential of +26.0 mV. After treatment, determination of zinc (Zn) concentration in the testes revealed desired impact of the exposure. Expression of several transcription factors and few steroidogenic enzyme genes in the treated testis showed significant downregulation than the control. Measurement of oxidative stress-related enzymes such as catalase, superoxide dismutase, and glutathione-S-transferase revealed substantial elevation in the testis of treated groups when compared to control. Histological analysis of testis exhibited dose-related response, defective lumen, and slow progression of spermatogenesis. Exposure of both the forms of Zn on TM3 Leydig cell culture displayed loss of adhesion, clumping with decreased viability, and a significant increase in the apoptotic cells. In addition, comet and intracellular reactive oxygen species (ROS) assays authenticated DNA damage upon treatment with a significant increase in ROS. Histological analysis after treatment withdrawal showed revival of testis in carp to rescue the effect. Thus, the present report highlights the adverse effect of Zn on the testis function in common carp as well as evident drastically toxic in in vitro cultures.

Gonadal transcriptome analysis of the common carp, Cyprinus carpio: Identification of differentially expressed genes and SSRs.

Common carp (Cyprinus carpio) is a world-wide freshwater fish of eutrophic waters. C. carpio, have various reproductive traits, including early sexual maturity, that may make them excellent, large, realistic, aquaculture model species. In the present work, de novo assembly of gonadal (testicular and ovarian) transcriptomes from juvenile common carp was performed to identify genes involved in gonadal development. A total of 81,757 and 43,257 transcripts with average lengths of 769 and 856 bp, were obtained from the immature testicular and ovarian transcriptomes, respectively. About 84,367 unigenes were constructed after removing redundancy involving representation of transcripts in both gonadal transcriptomes. Gene ontology (39,171 unigenes), clusters of orthologous group's analysis (6651 unigenes) and Kyoto encyclopedia of genes, and genomes automatic annotation server analysis (4783 unigenes) were performed to identify potential genes along with their functions. Furthermore, 18,342 (testis) and 8693 (ovary) simple sequence repeats were identified. About 298 differentially expressed genes were identified, of which 171 and 127 genes were up-regulated in testis and ovary, respectively. Quantitative real-time reverse transcription PCR was performed to validate differential expression of selected genes in testis and ovary. Nearly 809 genes related to reproduction were identified, sex-wise expression pattern of genes related to steroid synthesis, endocrine regulation, germ cell maintenance and others factors related to gonadal differentiation was observed, and expression analysis of nanos, ad4bp/sf-1, and gdf9 was performed. The present study identified certain important genes/factors involved in the gonadal development of C. carpio which may provide insights into the understanding of sex-differentiation and gonadal development processes.