The synthesis of broccoli sprout extract-loaded silk fibroin nanoparticles as efficient drug delivery vehicles: development and characterization.

Targeted drug delivery of biological molecules using the development of biocompatible, non-toxic and biodegradable nanocarriers can be a promising method for cancer therapy. In this study, silk fibroin protein nanoparticles (SFPNPs) were synthesized as a targeted delivery system for sulforaphane-rich broccoli sprout extract (BSE). The BSE-loaded SFPNPs were conjugated with polyethylene glycol and folic acid, and then their physicochemical properties were characterized via UV-Vis, XRD, FTIR, DLS, FE-SEM and EDX analyses. In vitro, the release profile, antioxidant and anticancer activities of NPs were also studied. The FE-SEM and DLS analyses indicated stable NPs with an average size of 88.5 nm and high zeta potential (-32 mV). The sulforaphane release profile from NPs was pH-dependent, with the maximum release value (70%) observed in simulated intestinal fluid (pH = 7.4). Encapsulation of BSE also decreased the release rate of sulforaphane from the capsules compared to free BSE. In vitro cytotoxicity of BSE and NPs on breast cancer cell lines (MCF-7) was concentration-dependent, and the IC50 for BSE and NPs were 54 and 210 µg ml-1, respectively. Moreover, the NPs demonstrated no appreciable cytotoxicity in normal mouse fibroblast (L929) cell lines. These results indicated that biocompatible NPs synthesized as controlled and long-term targeted drug delivery systems can be a potential candidate for breast cancer therapy.

Water deficit stress responses of monoterpenes and sesquiterpenes in different Iranian cultivars of basil.

Ocimum basilicum, a popular aromatic plant, contains aromatic terpenes of terpenoids with in vivo and in vitro verified cytotoxicity. Considering the characteristics and potential of its utilization, it would be attractive to reveal its regulation and biosynthesis, originally at the molecular level under water deficit stress. For this aim, for the first time, the gene encoding the enzyme involved in the end step of the MEP biosynthetic pathways (HDR) was cloned, and the accumulation ratio of linalool, germacrene D and γ-cadinene compounds as well as the expression trait of four critical genes (i.e., HDR, LinS, GerS, and GadS) was assessed under water deficit stress in three Iranian cultivars of basil. The highest value of linalool and γ-cadinene were detected for Cultivar 1 (Cult. 1) under mild stress (W1; 52.6 and 21.1%), while insignificant amounts were obtained for Cultivar 3 (Cult. 3). The germacrene D level of Cultivar 2 (Cult. 2) increased under severe and moderate water stresses as compared with mild water deficit stress. Apart from some expectation, all the studied genes demonstrated divergent transcription ratios under water deficit stress. Principal component analyses (PCA) showed that the relative water content (RWC) and HDR gene expression correlated significantly with essential oil components and gene expression in Cult. 1 and 2, which could represent an elevated demand for corresponding metabolites in the plant tissues. The present work elaborates on the regulation of the mentioned genes, and the results indicate that the production of terpenoids might be a drought stress-dependent and cultivar-dependent procedure.

A comparative study: Influence of various drying methods on essential oil components and biological properties of Stachys lavandulifolia.

The genus Stachys is a member of the Lamiaceae family. These are important medicinal plants which grow all over the world and are known for their flavoring and therapeutic effects and Stachys lavandulifolia is an endemic species of Iran. To acquire high-quality essential oil (EO), drying technique was implemented which is an essential part of this process. The present study designed to evaluate the influences of different drying techniques (fresh sample, shade, sunlight, freeze-drying, microwave, and oven-drying (40, 60, and 80°C) on EO yield and composition of S. lavandulifolia. The results indicated that the maximum EO yield was obtained by the shade-drying method. The main compounds found in the fresh samples were spathulenol, myrcene, ß-pinene, δ-cadinene, and α-muurolol, while spathulenol, cyrene, δ-cadinene, p-cymene, decane, α-terpinene, ß-pinene, and intermedeol were found to be the dominant compounds in the dry samples. Drying techniques were found to have a significant impact on the values of the main compositions, for example, monoterpene hydrocarbons such as α-pinene, ß-pinene, myrcene, and ß-phellandrene were significantly reduced by microwave drying, oven-drying (40, 60, and 80°C), and sunlight-drying methods. Drying techniques increased the antioxidant activity of S. lavandulifolia EOs especially those acquired by freeze-drying with the half-maximal inhibitory concentration (IC50) values 101.8 ± 0.8 mg/ml in DPPH assay and 315.2 ± 2.1 mg/ml in decreasing power assay. As a result, shade-, sun-, and oven-drying (40°C) were found to be the most important techniques for attaining maximum yields of EO.

In Vitro and In Vivo Determinations of The Anti-GDNF Family Receptor Alpha 1 Antibody in Mice by Immunochemistry and RT-PCR.

BACKGROUND: The glial cell-derived neurotrophic factor (GDNF) family plays essential roles in the maintenance, growth, regulatory and signalling pathways of spermatogonial stem cells (SSCs). In this study, we analysed the expression of anti-GDNF family receptor alpha 1 antibody (GFRa1) by immunohistochemistry (IHC), immunocytochemistry (ICC), Fluidigm real-time polymerase chain reaction (RT-PCR) and flow cytometry analyses. MATERIALS AND METHODS: In this experiment study, ICC, IHC, Fluidigm RT-PCR and flow cytometry were used to analyse the expression of the germ cell marker GFRa1 in testis tissue and SSC culture. RESULTS: IHC analysis showed that there were two groups of GFRa1 positive cells in the seminiferous tubules based on their location and expression shape - a small round punctuated shape on the basal compartment donut shape and a C-shaped expression located between the basal and the luminal compartments of the seminiferous tubules. OCT4 and PLZF positive cells may have similar patterns of expression as the first group. Assessment of the seminiferous tubule sections demonstrated that about 27% of the SSCs were positive for GFRa1. Fluidigm RT-PCR confirmed the significant expression (P<0.001) of GFRa1 in the SSCs compared to testicular stromal cells (TSCs). Flow cytometry analysis demonstrated that about 75% of the isolated SSCs colonies were positive for GFRa1. CONCLUSION: The results indicated that GFRa1 had a specific expression pattern both in vivo and in vitro. This finding could be helpful for understanding the proliferation, maintenance and signalling pathways of SSCs, and differentiation of meiotic and haploid germ cells.

Fatty acid profile and in vitro biological properties of two Rosacea species (Pyrus glabra and Pyrus syriaca), grown as wild in Iran.

The high demands for the consumption of edible oils have caused scientists to struggle in assessing wild plants as a new source of seed oils. Therefore, in this study, the oil yield, fatty acid and tocopherol compositions, antioxidant and antibacterial activities of the oils obtained from Iran's two endemic plants (Pyrus glabra and Pyrus syriaca) were investigated. The obtained oil yields from the P. glabra and P. syriaca seeds were 33 ± 0.51 and 26 ± 0.28 w/w%, respectively. Oleic acid (C18:1) with the amount of 49.51 ± 1.05% was the major fatty acid in the P. glabra oil, while the main fatty acids in the P. syriaca seed oil belonged to linoleic acid (C18:2) and oleic acid (C18:1) with the amounts of 46.99 ± 0.37 and 41.43 ± 0.23%, respectively. The analysis of tocopherols was done by HPLC, and the results indicated that the P. glabra and P. syriaca seed oils were rich in α-tocopherol (69.80 ± 1.91 and 45.50 ± 1.86 mg/100 g oil, respectively), constituting 86.24 and 89.01% of total detected tocopherols, respectively. The study on the reducing capacity of the oils indicated that the P. glabra oil had more reducing capacity than the P. syriaca oil. Moreover, the antioxidant activity of the P. glabra seed oil (43.4 ± 0.7 µg/ml) was higher than the P. syriaca seed oil (46.3 ± 1.2 µg/ml). Also, the investigation of the antibacterial activities indicated that the P. glabra and P. syriaca oils have an inhibitory effect on the studied bacteria. The results indicate that the oils of these plants can be appropriate sources of plant oils which can act as natural antibacterial agents.

Investigation of VASA Gene and Protein Expression in Neonate and Adult Testicular Germ Cells in Mice In Vivo and In Vitro.

OBJECTIVE: We aimed to examine the expression levels of the VASA gene and protein in testis sections of neonate and adult mice as well as testicular cell cultures. MATERIALS AND METHODS: In this experimental study, in order to investigate the expression of this germ cell marker gene in more detail, we analyzed the expression of VASA by immunocytochemistry, immunohistochemistry and fluidigm reverse transcription-polymerase chain reaction (RT-PCR). RESULTS: The immunohistochemical assays showed that the VASA protein was exclusively expressed in germ cells in the seminiferous tubules of the neonate and adult testis and not in somatic cells. VASA was not detectable in PLZF positive spermatogonial stem cells (SSCs), was weakly expressed in proliferating spermatogonia, and became abundant in spermatocytes and round spermatozoa. Counting VASA-positive cells in the seminiferous tubules of the neonate and adult testis depicted significant higher expression (P<0.05) of VASA in the adult testis in comparison to its neonate counterpart. SSC colonies were established in vitro after digestion of the testis and characterized by immunocytochemistry for CD90 and stage-specific embryonic antigens 3 (SSEA3). Immunocytochemistry confirmed that in contrast to the not detectable signal in vivo, VASA protein was strongly localized in the cytoplasm of both neonate and adult mouse SSCs under in vitro conditions. The results of Fluidigm RT-PCR revealed a significant higher expression of the germ cell gene VASA in adult SSCs in comparison to neonate SSCs in cell culture (P<0.05). CONCLUSION: The VASA protein is, therefore, an extremely specific marker of testicular germ cell differentiation in vivo and mostly expressed in the adult testis in spermatocytes and round spermatids. The immunohistochemical signal in spermatogonia is very low. So, PLZF positive SSCs are negative for VASA in vivo, while in contrast, once isolated from the testicular niche VASA is also strongly expressed in SSCs under in vitro conditions.

Incorporation of plant growth regulators into the priming solution improves sugar beet germination, emergence and seedling growth at low-temperature.

In a series of experiments, impact of inclusion of plant growth regulators into the KNO3 priming solution on low temperature seed germination, emergence percentage and seedling growth of sugar beet was investigated. Seeds were primed in 3% KNO3 solution for 6 days at 25 degrees C in darkness containing one of the following: 0, 0.05, 0.1, 0.5, or 1 mM acetyl salicylic acid (ASA) or 1, 3, 5 or 10 microM methyl jasmonated (MeJA). A non-primed treatment was also included in the experiment. Priming seeds in the presence or absence of plant growth regulators in general improved final germination percentage (FGP), germination rate (G50) and germination synchrony (G10-90) at 15 degrees C compared with non-primed seeds which had an FGP of 42%, G50 of 11.3 days and G10-90 of 11.7 days. Priming seeds in KNO3 solution containing 0.05 mM of ASA resulted in the highest germination percentage (89%), fastest germination rate (G50 = 5.3 days) and the most synchronous germination (G10-90 = 10.7 days). Emergence percentages were the highest for the seeds primed in the presence of 0.05 mM ASA (83%) and 3 microM MeJA (81%) while non-primed seeds had an emergence percentage of 40%. Fastest emergence rate (E50) were also obtained from seeds primed in KNO3 supplemented with 3 microM MeJA (E50 = 14.4 days) and 0.05 mM ASA (E50 = 14.4 days). Shoot fresh and dry weight of seedlings were significantly affected by treatments and priming in the presence of 0.05 mM ASA resulted in highest seedling shoot fresh and dry weight. These results indicate that priming seeds in 0.05 mM of ASA or 3 microM MeJA incorporated into the KNO3 solution can be more effective than KNO3 alone to improve low temperature germination performance of seeds and subsequent seedling growth.