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BACKGROUND: Green synthesized nanoparticles have been earmarked for use in nanomedicine including for the development of better anticancer drugs. OBJECTIVE: The aim of this study was to undertake biochemical evaluation of anticancer activities of green synthesized silver nanoparticles (AgNPs) from ethanolic extracts of fruits (AgNPs-F) and leaves (AgNPs-L) of Annona muricata. METHODS: Previously synthesized silver nanoparticles were used for the study. The effects of the AgNPs and 5-Fluorouracil were studied on PC3, HeLa and PNT1A cells. The resazurin, migration and colonogenic assays as well as qRT-PCR were employed. RESULTS: The AgNPs-F displayed significant antiproliferative effects against HeLa cells with an IC50 of 38.58μg/ml and PC3 cells with an IC50 of 48.17μg/ml but selectively spared normal PNT1A cells (selectivity index of 7.8), in comparison with first line drug 5FU and AgNPs-L whose selectivity index were 3.56 and 2.26 respectively. The migration assay revealed potential inhibition of the metastatic activity of the cells by the AgNPs-F while the colonogenic assay indicated the permanent effect of the AgNPs-F on the cancer cells yet being reversible on the normal cells in contrast with 5FU and AgNPs-L. CASP9 was significantly over expressed in all HeLa cells treated with the AgNPs-F (1.53-fold), AgNPs-L (1.52-fold) and 5FU (4.30-fold). CXCL1 was under expressed in HeLa cells treated with AgNPs-F (0.69-fold) and AgNPs-L (0.58-fold) and over expressed in cells treated with 5FU (4.95-fold), but the difference was not statistically significant. CXCR2 was significantly over expressed in HeLa cells treated with 5FU (8.66-fold) and AgNPs-F (1.12-fold) but under expressed in cells treated with AgNPs-L (0.76-fold). CONCLUSIONS: Here we show that biosynthesized AgNPs especially AgNPs-F can be used in the development of novel and better anticancer drugs. The mechanism of action of the AgNPs involves activation of the intrinsic apoptosis pathway through upregulation of CASP9 and concerted down regulation of the CXCL1/ CXCR2 gene axis.

Animal mummies from ancient Egypt are not extensively studied in comparison with human mummies. Dogs mummies were very rare in ancient Egypt, and at the same time were not adequately studied by researchers from conservation point of view. Dog mummy in the present study is dated back to late period (664−333 BC.) and is preserved in the agriculture museum (Egypt). It suffers from different aspects of deterioration such as dirt, holes, wrinkles, missing tissue, accumulated dust, etc. This study aims to evaluate the preservation state of the mummy and explain the deterioration mechanisms to help the conservators later to choose the appropriate materials and methods for the conservation treatment. The analytical techniques used were Auto-Cad, scanning electron microscope (SEM), determination of moisture content, pH measurement, amino acid analysis (AAA), fourier transform infrared (FTIR), portable X-ray fluorescence (XRF), inductively coupled plasma mass spectrometry (ICP-MS) and X-ray diffracion analysis. The results obtained showed different forms of deterioration. Moisture content measurement proved that the mummy was dehydrated. The pH measurement of the mummy components is in acid level. FTIR and amino acid analysis proved that the mummy have been exposed to oxidation process. The analysis by XRF, ICP-MS and XRD proved that bone of the mummy suffered from deterioration.

Introduction: The biological green synthesis of nanoparticles via nanobiotechnology processes have a significant potential to boost nanoparticles production without the use of harsh, toxic, and expensive chemicals commonly used in conventional physical and chemical processes. Annona muricata, a tropical plant belonging to family Annonaceae is one of the most used plants in folk medicine because of its many medicinal uses and therefore presents a strong candidate for use in green synthesis. Aims: The aim of this study was to optimize a method for the synthesis of Silver Nanoparticles (AgNPs) from ethanolic extracts of leaves of Annona muricata as well as to characterize the green synthesized AgNPs. Methodology: AgNPs were synthesized from Annona muricata leaves using AgNO3 solution. The AgNPs were characterized using spectroscopy and microscopy techniques. Results: The formed AgNPs had an absorption maximum at 429 nm using UV–Visible spectroscopy and were stable under different pH, temperature, and storage conditions. Fourier transform infrared analysis revealed the different functional groups responsible for the synthesis and stabilization of the AgNPs. Scanning electron microscopy analysis revealed a spherical nature of the synthesized AgNPs. Energy dispersive x-ray spectroscopy analysis showed presence of Ag, O, and Cl with Ag having the highest composition at 60%. X-Ray Diffraction and Dynamic Light Scattering revealed a crystalline nature of AgNPs with an average size of 87.36 nm and a polydispersity index of 0.16 respectively. Transmission Electron Microscopy analysis further confirmed the crystalline and spherical nature of the AgNPs. Conclusion: In this article, an efficient, eco-friendly and low-cost method for the synthesis and recovery of stable AgNPs using Annona muricata leaves ethanolic extracts as both a reducing and capping agent has been reported for the first time. The synthesized AgNPs could be promising candidates for many biomedical, clinical, engineering, and polymer applications.

Green synthesis of nanoparticles from plant materials opens a new scope in nanobiotechnology and discourages the use of expensive toxic chemicals. The aim of this study was to develop and optimize a method for the synthesis of Silver Nanoparticles (AgNPs) from ethanolic extracts of fruits of Annona muricata as well as to characterize the green synthesized AgNPs. AgNPs were synthesized via AgNO3 solution. The AgNPs were characterized using spectroscopy and microscopy techniques. The formed AgNPs had an absorption maximum of 427 nm and were stable under different temperature, pH and storage conditions. Fourier Transform Infrared Resorption spectroscopy revealed the functional groups responsible for the synthesis and stabilization of the AgNPs. Scanning Electron Microscopy analysis revealed a spherical nature of the AgNPs. Energy Dispersive X-Ray spectroscopy showed presence of Ag, Cl, Ca, and Si with Ag having the highest composition at 80%. X-ray diffraction and dynamic light scattering revealed a crystalline nature of AgNPs with an average size of 60.12 nm and a polydispersity index of 0.1235 respectively. Transmission Electron Microscopy analysis further confirmed the crystalline and spherical nature of the AgNPs. In this article, an efficient, eco-friendly and low-cost method for the synthesis and recovery of stable AgNPs using ethanolic extracts of Annona muricata fruits as both reducing and capping agents has been reported. The synthesized AgNPs could have many biomedical and clinical applications.

Dioecism has always been an issue in many plant species with its numerous disadvantages, especially in woody trees such as date palms. As one of the most important crops in the Middle Eastern countries, researchers are having problems identifying of sex of the plant in its early stages of development. Hence, proper population stands in the male: female ratio for maintenance is almost impossible in the field for better production. In this study, sex determination of date palm (Phoenix dactilyfera L.) were identified in regions of the Y chromosome (Date-SRY) gene, the pivotal gene that initiates sex determination, using a new technique and thus an economically desirable objective, which will significantly impact profits in seed based cultivations. Partial sequences of the Date-SRY were taken and amplified by nested polymerase chain reaction (PCR). According to the results, the exact sex of date palm was identified in all the tested plants, while amplified regions of the Date-SRY gene closely matched with the human and papaya sequences. In addition, a primer pair was designed to amplify the sequences of the SRY-date gene with confidence that it will identify male date palms. These primer sequences include SRY-date Forward 5′- cggccctctaagtatctgtgcgcaacg-3′ (SRY-date F) and the SRY-date Reverse 5′- gtttgcacttcgaagcagag-3′ (SRY-date R). The complete sequence of the DNA has been registered and deposited in GenBank (BankIt1598036 DPSRY1 KC577225 thenKJ873056). Keywords: Date palm, Dioecious plants, Female, Male, Sex determination, SRY gene sequence

This study aims to highlight the antioxidant and cytotoxic effects of Cichorium intybus L.. The cytotoxic activity for cancer cell lines was evaluated by MTT assay. Total phenol and flavonoid conte...

Mummification was considered one of the most important processes used for the preservation of the body through ages in ancient Egypt for long times. Some natural materials were used to achieve the preservation goal. Some of these materials are natural resins which can be used alone as separate or as a mixture with others such as fat, wax, and oils. Some environmental factors (interior or exterior) were affecting these mixture components and lead to its degradation. This study aims to identify the chemical composition by gas chromatography mass spectrometry (GC/MS) and explain the deterioration mechanism of some mixture materials used in the preservation process of Late Period mummy. The results revealed that the preservative mixture samples contained resins pine pitch and mastic mixed with some biochemical substances such as fat and wax. The results also showed that the packing of the mummy’s remains in plastic bags led to the presence of phthalate compounds as contaminants. Moreover, the degradation compounds of acetonitrile and 2-amino-n-isopropylbenzamide in the ancient tissue have been demonstrated with the sample taken from the cranial cavity. Additionally, phenanthrene compounds and its derivatives were identified, and this may be due to the use of pine pitch in the embalming mixture.

Liver is the key organ (detoxifier) that gets injured by reactive oxygen species (ROS) which deplete the body's stores of anti-oxidants. ROS is present in all chronic liver diseases leading to oxidative stress. Consumption of foods high in anti-oxidants replenishes and supplies the body with important anti-oxidants which break down ROS lowering inflammatory responses and preventing liver damage. In this work, clove and honey, which are known to be rich in anti-oxidants extracts were evaluated for their potential to breakdown ROS, Reactive nitrogen species (RNS) and lower oxidative stress in rats. Purified extracts were tested in vitro for ROS scavenging by 1,1-diphenyl-2-picrylhydrazyl (DPPH) and in vivo using CCl4-induced liver injury model. A total of 50 experimental rats were assigned to 5 groups each consisting of 10 animals and treated as follows; 1. CCl4 group: fed on CCl4 in olive oil 2. honey group: fed CCl4 then honey extract, 3. clove group: CCl4 then clove extract, 4. double negative control group: rats fed on standard diet and 5 Negative control: fed olive oil, At 10 weeks, liver histopathology was done and sera collected for liver and kidney function tests. CCl4 -only treated rats showed fibroblastic cells proliferation and inflammatory cells infiltration which was absent in the control animals. Clove extract did not exhibit fibroblastic cells proliferation at all. Both clove and honey significantly reduced cytokines (TGF-β, TNF-α and EGF) and liver function enzymes'(ALT, AST and GGT) activity (58.5,56; 119,121 and 6.1; 6.3) compared to the CCl4 group (97; 191 and 10.1). Clove highly inhibited DPPH (77.2%) compared to honey (6.5%) and conferred better liver protection against CCl4. These findings indicate that clove and honey contain compounds that have anti-oxidant activity with the ability to reverse liver fibrosis. Frequent consumption of these foods may lower liver damage and prevent development of some liver diseases. Public health education on nutritional benefits of these foods is strongly encouraged. Keywords: Clove, Honey, Liver, CCl4, Antioxidant, ROS

The resins and other biochemical substances in mixtures have an important role in achieving the preservation goal for mummification process. The identification of organic mixtures is vital in the conservation field of the unwrapped mummies, but their identification is sometimes difficult because of improper conditions, which can affect their structure and need good investigation techniques for their identification. Some endogenous or exogenous factors can cause some aspects of deterioration to organic mixtures such as cracks, color change, and chemical alteration. This study aims to evaluate the materials used for the preservation of the body in order for them to be re-used in the near future for the preservation processes on the unwrapped mummies in Egyptian museums and storehouses. Samples of organic mixtures were taken from an ancient Egyptian mummy dated to the Late Period (664–333 B.C.). A digital USB microscope and a scanning electron microscope (SEM) were used to identify the application method of the preservative mixtures used during the mummification process and to demonstrate the morphology of surface alteration on the body. Fourier transform infrared (FTIR) was used to detect the chemical changes. A portable X-ray fluorescence (XRF) was used to identify elements. The results revealed that the investigation by microscopes showed cracks in the mixtures poured inside the cranial cavity, but the application of these mixtures was by brush on the outer surface layer of the body. FTIR analysis revealed that a mixture of oil, animal fat, and resinous materials such as mastic and pine were present in the studied samples. Moreover, these samples were prone to oxidation processes through time. XRF analysis showed that the main chemical elements of the samples were calcium, potassium, phosphorus, chlorine, sulfur, and silicon and trace chemical elements such as iron, zinc, copper, and lead, which are in the chemical composition of pine and mastic resins.

The genus Nostoc is a valuable source of a wide spectrum of natural products, which have many different biological activities such as antioxidant, anticancer, anti-HIV, antimalarial, antifungal and antimicrobial drugs. The current work was aimed to evaluate the biological activities of Nostoc muscorum microalga cultivated on different waste water samples (domestic, agricultural and industrial waste water). The algae was cultivated in different kinds of waste water; sterilized BG11 media, non-sterilized treated sewage waste water media (NS-TSW), sterilized treated sewage waste water media (S-TSW), 50% BG11 + 50% sterilized TSW media (50% S-TSW), non-sterilized industrial waste water media (NS-IW), sterilized industrial waste water media (S-IW), 50% BG11 + 50% sterilized industrial waste water media (50% S-IW), non-sterilized agriculture waste water media (NS-AW), sterilized agriculture waste water media (S-AW), and 50% BG11 + 50% sterilized AW media (50% S-AW). The experiment was conducted in triplicate and cultures were incubated at 25 ± 1 °C under continuous shaking (150 rpm) and illumination (2000 lx) for 15 days. The pigments content and biological activity include antioxidant (using diphenyl picryl hydrazyl and 2,2-azino-bis ethylbenzthiazoline-6-sulfonic acid radical methods), antiviral activity against Herpes simplex virus and the cytotoxicity for aqueous extracts in vivo and in vitro assays were determined. The given results reported that use of waste water as media for cultivation of microalgae is a suitable and inexpensive method when compared with ordinary cultivation methods. Also We can conclude that the algal species has high ability to produce active ingredients used as antioxidants and antivirals especially from algae cultivated in S-TSW, 50% S-TSW, NS-AW and 50% S-AW without any toxicity in mice.

Annona muricata (A. muricata) is a tropical plant species belonging to family Annonaceae and known for its many medicinal uses. This review focuses on the research history of its traditional uses, phytochemicals, pharmacological activities, toxicological aspects of the extracts and isolated compounds, as well as the in vitro propagation studies with the objective of stimulating further studies on this plant for human consumption and treatment. A. muricata extracts have been identified in tropical regions to traditionally treat diverse conditions ranging from fever to diabetes and cancer. More than 200 chemical compounds have been identified and isolated from this plant, the most important being alkaloids, phenols and acetogenins. Using in vitro studies, its extracts and phytochemicals have been characterized as antioxidant, anti-microbial, anti-inflammatory, insecticidal, larvicidal, and cytotoxic to cancer cells. In vivo studies have revealed anxiolytic, anti-stress, anti-inflammatory, immunomodulatory, antimalarial, antidepressant, gastro protective, wound healing, hepato-protective, hypoglycemic, anticancer and anti-tumoral activities. In silico studies have also been reported. In addition, clinical studies support the hypoglycemic as well as some anticancer activities. Mechanisms of action of some pharmacological activities have been elucidated. However, some phytochemical compounds isolated from A. muricata have shown a neurotoxic effect in vitro and in vivo, and therefore, these crude extracts and isolated compounds need to be further investigated to define the magnitude of the effects, optimal dosage, and mechanisms of action, long-term safety, and potential side effects. Additionally, more clinical studies are necessary to support the therapeutic potential of this plant. Some studies were also found to have successfully regenerated the plant in vitro, but with limited success. The reported toxicity notwithstanding, A. muricata extracts seem to be some of the safest and promising therapeutic agents of the 21st century and beyond that need to be studied further for better medicinal formulations and diseases management.

Sodic-alkalinity may be more deleterious to plant growth than salinity. The objectives of this study were to determine whether 5-aminolevulinic acid (ALA: an essential precursor for chlorophyll biosynthesis) foliar application could improve the sodic-alkaline resistance of Swiss chard (Beta vulgaris L. subsp. cicla) by regulating water uptake, ionic homeostasis, photosynthetic capacity, and antioxidant metabolism. Eight-week-old uniform plants were grown in nutrient medium without and with a sodic-alkaline regime generated by a mixture of NaHCO3 and Na2CO3 (NaHCO3 : Na2CO3 = 9:1 molar ratio) for 12 d, and leaves were sprayed daily with water or ALA. The Na+ and ALA concentrations were gradually increased to 60 mM and 120 μM, respectively. ALA foliar application alleviated the physiological damage from sodic-alkalinity, as reflected by the increases in plant dry weight, relative growth rate, chlorophyll, Mg2+ concentration, and the decrease in Na+ concentration. However, ALA foliar application did not change the water uptake capacity or the concentration of K+, Fe3+, and endogenous ALA in leaf tissues under sodic-alkaline conditions. ALA foliar application effectively mitigated damage from sodic-alkalinity because of the increased activity of antioxidant enzymes (catalase and guaiacol peroxidase), particularly superoxide dismutase activity, which was maintained at the same level as for control plants. These results suggest that ALA foliar application alleviated sodic-alkaline stress mainly owing to its antioxidant capacity, and superoxide dismutase has the main responsibility for reducing oxidative stress in Swiss chard.

The genes underlying rhg1 lie at a sometimes dominant sometimes co-dominant locus, necessary for resistance to all Hg types of the soybean (Glycine max (L.) Merr.) cyst nematode (Heterodera glycines). Genomic research identified; nucleotide changes within a candidate gene encoding a receptor like kinase (RLK) that were capable of altering root development and thereby part of the resistance to Hg types 0 (race 3); changes in a laccase that are capable of altering cyst development; and genes underlying changes in membrane biology. This set of three genes are subject to co-selection with a modifier locus on another linkage block. Root development is slowed in the resistant seedling and results in end of season yield loss when SCN is not present. However, in the presence of SCN resistant seedling roots grow just as vigorously as the now slower growing parasitized susceptible roots and therefore show little loss to SCN parasitism. In some genotypes but not others the RLK can act alone to confer resistance. Functional paralogs of the three gene cluster have been found on other linkage groups including A1, B1, G, and O and these can be functional in different sources of resistance like G. soja, PI 437654 and PI438489B. At rhg1 the allele differences change the structure, interacting partners and activity of the LRR protein and the laccase. The changes between the alleles result in about 30 other proteins (judged by 2 Dgels), 112 metabolites (by FTICRMS) and 8 metabolites (by GCMS) to increase in abundance in roots during SCN infection in the resistant NILs. Understanding the basis of root stunting by resistance alleles will be used to improve methods for developing new nematode resistant soybean cultivars that do not suffer from the yield suppression and low seed germination rates of existing cultivars.

In this data article, data obtained from an efficient, eco-friendly and low-cost method for the synthesis and recovery of Silver nanoparticles (AgNPs) using ethanolic extracts of Annona muricata fruits and leaves as reducing, stabilizing and capping agents has been reported. 99.7% pure silver nitrate was used as the inorganic ion source. The data was obtained using different spectroscopic and microscopic techniques. The data is presented in form of images, Microsoft excel sheets, graphs,.raw files,.dpt files, PDF files, among others. Methods of analysis and interpretation of the data have also been presented. The data can be most useful to researchers, research students, industrialists and academicians to acquire knowledge on the green synthesis of AgNPs and related applications. The data is deposited in the Mendeley Data Repository as two independent datasets accessible at https://doi.org/10.17632/jkj2x782wh.1 Gavamukulya et al., 2019 and https://doi.org/10.17632/f4mb6b488n.1 Gavamukulya et al., 2019. Keywords: Annona muricata, Green synthesis, Silver nanoparticles (AgNPs), UV/VIS, FTIR, XRD

The aim of the present study was to assess in vitro the antiradical and antioxidant activities of successive extracts and semi-purified fractions from Rumex vesicarius L. In the present work, three extracts (n-Hexane, ethyl acetate and methanol) and 22 column fractions of methanolic extract (as promising extract) were evaluated against 2,2-diphenyl- 1-picrylhydrazyl (DPPH•) and 2,2-azinobis (3-ethylbenzothiazoline- 6-sulfonic acid) (ABTS) radical scavenging methods as antiradical and antioxidant activities compared with Butylated hydroxytoluene (BHT) as synthetic standard and silver nanoparticles of methanolic extract (Ag-NPs-Me), in addition to analysis of chemical constituents of extract and fraction using Gas chromatography–mass spectrometry (GC-MS). The obtained results revealed that, both methods go parallel showing that the concentration of extract and incubation time are dependent and proportional with phenolic compounds concentration. Absolute methanol extract recorded the highest antioxidant activity when compared with the other crude extracts with 79.3 and 78.8% against DPPH and ABTS respectively when compared with BHT as synthetic standard (89.4 and 89.9%) against DPPH and ABTS respectively. Calculation of the antiradical activity units showed the highest values of methanolic extract and its promising fraction (No. 12) after 300 seconds (5 minutes) comparing with antioxidant activity (30 min). Also, the antioxidant activity increased with synthetic Ag-NPs-Me when compared with methanolic extract by (IC50= 53.9 and 74.6 µg/ml respectively). Thus, the GC-MS analysis of successive extracts of R. vesicarius L showed a highly complex profile, containing approximately 24 different components. One pure compound was identified from fraction No. 12. The identified compound was l-(+)- ascorbic acid 2, 6-dihexadecanoate. The data also revealed presence of closely similar antioxidant activities in methanolic extract or its pure compounds with BHT when mixed at different proportions. From the obtained results it could be concluded that R. vesicarius methanolic extracts and fractions can be extensively used in the production of potential antioxidant, antiradical and AgNPs-Me for biomedical application on the consumer’s health.

Spermatogenesis is a complex and highly regulated process by which spermatogonial stem cells differentiate into spermatozoa. To better understand the molecular mechanisms of the process, the Cre/loxP system has been widely utilized for conditional gene knockout in mice. In this study, we generated a transgenic mouse line that expresses Cre recombinase under the control of the 2.5 kbp of the Prolactin family 3, subfamily b, member 1 (Prl3b1) gene promoter (Prl3b1-cre). Prl3b1 was initially reported to code for placental lactogen 2 (PL-2) protein in placenta along with increased expression toward the end of pregnancy. PL-2 was found to be expressed in germ cells in the testis, especially in spermatocytes. To analyze the specificity and efficiency of Cre recombinase activity in Prl3b1-cre mice, the mice were mated with reporter R26GRR mice, which express GFP ubiquitously before and tdsRed exclusively after Cre recombination. The systemic examination of Prl3b1-cre;R26GRR mice revealed that tdsRed-positive cells were detected only in the testis and epididymis. Fluorescence imaging of Prl3b1-cre;R26GRR testes suggested that Cre-mediated recombination took place in the germ cells with approximately 74% efficiency determined by in vitro fertilization. In conclusion, our results suggest that the Prl3b1-cre mice line provides a unique resource to understand testicular germ-cell development. genesis 54:389-397, 2016. © 2016 Wiley Periodicals, Inc.

The merging of high-throughput gene expression techniques, such as microarray, in the screening of natural products as anticancer agents, is considered the optimal solution for gaining a better understanding of the intervention mechanism. Red yeast rice (RYR), a Chinese dietary product, contains a mixture of hypocholesterolemia agents such as statins. Typically, statins have this effect via the inhibition of HMG‑CoA reductase, the key enzyme in the biosynthesis of cholesterol. Recently, statins have been shown to exhibit various beneficial antineoplastic properties through the disruption of tumor angiogenesis and metastatic processes. Mevinolin (MVN) is a member of statins and is abundantly present in RYR. Early experimental trials suggested that the mixed apoptotic/necrotic cell death pathway is activated in response to MVN exposure. In the current study, the cytotoxic profile of MVN was evaluated against MCF‑7, a breast cancer‑derived cell line. The obtained results indicated that MVN‑induced cytotoxicity is multi‑factorial involving several regulatory pathways in the cytotoxic effects of MVN on breast cancer cell lines. In addition, MVN‑induced transcript abundance profiles inferred from microarrays showed significant changes in some key cell processes. The changes were predicted to induce cell cycle arrest and reactive oxygen species generation but inhibit DNA repair and cell proliferation. This MVN‑mediated multi‑factorial stress triggered specific programmed cell death (apoptosis) and DNA degradation responses in breast cancer cells. Taken together, the observed MVN‑induced effects underscore the potential of this ubiquitous natural compound as a selective anticancer activity, with broad safety margins and low cost compared to benchmarked traditional synthetic chemotherapeutic agents. Additionally, the data support further pre‑clinical and clinical evaluations of MVN as a novel strategy to combat breast cancer and overcome drug resistance.

Organic fractions and extracts of willow (Salix safsaf) leaves, produced by sequential solvent extraction as well as infusion and decoction, exhibited anticancer potencies in four cancerous cell lines, including breast (MCF-7), colorectal (HCT-116), cervical (HeLa) and liver (HepG2). Results of the MTT assay revealed that chloroform (CHCl3) and ethyl acetate (EtOAc)-soluble fractions exhibited specific anticancer activities as marginal toxicities were observed against two non-cancerous control cell lines (BJ-1 and MCF-12). Ultra-high-resolution mass spectrometry Q-Exactive&trade, HF Hybrid Quadrupole-Orbitrap&trade, coupled with liquid chromatography (UHPLC) indicated that both extracts are enriched in features belonging to major phenolic and purine derivatives. Fluorescence-activated cell sorter analysis (FACS), employing annexin V-FITC/PI double staining indicated that the observed cytotoxic potency was mediated via apoptosis. FACS analysis, monitoring the increase in fluorescence signal, associated with oxidation of DCFH to DCF, indicated that the mechanism of apoptosis is independent of reactive oxygen species (ROS). Results of immunoblotting and RT-qPCR assays showed that treatment with organic fractions under investigation resulted in significant up-regulation of pro-apoptotic protein and mRNA markers for Caspase-3, p53 and Bax, whereas it resulted in a significant reduction in amounts of both protein and mRNA of the anti-apoptotic marker Bcl-2. FACS analysis also indicated that pre-treatment and co-treatment of human amniotic epithelial (WISH) cells exposed to the ROS H2O2 with EtOAc fraction provide a cytoprotective and antioxidant capacity against generated oxidative stress. In conclusion, our findings highlight the importance of natural phenolic and flavonoid compounds with unparalleled and unique antioxidant and anticancer properties.

Plant response to salt stress and the mechanism of salt tolerance have received major focus by plant biology researchers. Biotic stresses cause extensive losses in agricultural production globally, but abiotic stress causes significant increase in the methylglyoxal (MG) level of GlyoxalaseI (Gly I). Identification of salt-tolerant genes when characterizing their phenotypes will help to identify novel genes using polymerase chain reaction (PCR) to amplify the DNA coding region for glyoxalase I. This method is specific, requiring only genomic DNA and two pairs of PCR primers, and involving two successive PCR reactions. This method was used rapidly and easily identified glyoxalase I sequences as salt-tolerant genes from Jojoba (Simmondsia chinensis (Link) Schneider). In the present study, the glyoxalase I gene was isolated, amplified by PCR using gene-specific primers and sequenced from the jojoba plant, then compared with other glyoxalase I sequences in other plants and glyoxalase I genes like in Brassica napus, ID: KT720495.1, Brassica juncea ID: Y13239.1, Arachis hypogaea, ID: DQ989209.2, and Arabidopsis thaliana L, ID: AAL84986. The structural gene of glyoxalase I, when sequenced and analyzed, revealed that the uninterrupted open reading frame (ORF) of jojoba Gly I (Jojo-Gly I) spans 775 bp, corresponding to 185 amino acid residues, and shares 45.2% amino acid sequence identity to jojoba (Jojo-Gly I). The cloned ORF, in a multicopy constitutive expression plasmid, complemented the Jojo-Gly I, confirming that the encoded Jojo-Gly I in jojoba showed some homology with other known glyoxalase I sequences of plants.

The transcription factor MAFB is an important regulator of the development and differentiation of various organs and tissues. Previous studies have shown that MAFB is expressed in embryonic and adult mouse testes and is expected to act as the downstream target of retinoic acid (RA) to initiate spermatogenesis. However, its exact localization and function remain unclear. Here, we localized MAFB expression in embryonic and adult testes and analyzed its gene function using Mafb-deficient mice. We found that MAFB and c-MAF are the only large MAF transcription factors expressed in testes, while MAFA and NRL are not. MAFB was localized in Leydig and Sertoli cells at embryonic day (E) 18.5 but in Leydig cells, Sertoli cells, and pachytene spermatocytes in adults. Mafb-deficient testes at E18.5 showed fully formed seminiferous tubules with no abnormal structure or differences in testicular somatic cell numbers compared with those of control wild-type mice. Additionally, the expression levels of genes related to development and function of testicular cells were unchanged between genotypes. In adults, the expression of MAFB in Sertoli cells was shown to be stage specific and induced by RA. By generating Mafbfl/fl CAG-CreER™ (Mafb-cKO) mice, in which Cre recombinase was activated upon tamoxifen treatment, we found that the neonatal cKO mice died shortly upon Mafb deletion, but adult cKO mice were alive upon deletion. Adult cKO mice were fertile, and spermatogenesis maintenance was normal, as indicated by histological analysis, hormone levels, and germ cell stage-specific markers. Moreover, there were no differences in the proportion of seminiferous stages between cKO mice and controls. However, RNA-Seq analysis of cKO Sertoli cells revealed that the down-regulated genes were related to immune function and phagocytosis activity but not spermatogenesis. In conclusion, we found that MAFB is dispensable for fetal testis morphogenesis and spermatogenesis maintenance in adult mice, despite the significant gene expression in different cell types, but MAFB might be critical for phagocytosis activity of Sertoli cells.

Pan African University Institute for Basic Sciences, Technol¬ogy and Innovation, Cairo University, Makerere University, Uganda Directorate of Government Analytical Laboratory

Malaria causes about half a million deaths annually, with Plasmodium falciparum being responsible for 90% of all the cases. Recent reports on artemisinin resistance in Southeast Asia warrant urgent discovery of novel drugs for the treatment of malaria. However, most bioactive compounds fail to progress to treatments due to safety concerns. Drug repositioning offers an alternative strategy where drugs that have already been approved as safe for other diseases could be used to treat malaria. This study screened approved drugs for antimalarial activity using an in silico chemogenomics approach prior to in vitro verification. All the P. falciparum proteins sequences available in NCBI RefSeq were mined and used to perform a similarity search against DrugBank, TTD and STITCH databases to identify similar putative drug targets. Druggability indices of the potential P. falciparum drug targets were obtained from TDR targets database. Functional amino acid residues of the drug targets were determined using ConSurf server which was used to fine tune the similarity search. This study predicted 133 approved drugs that could target 34 P. falciparum proteins. A literature search done at PubMed and Google Scholar showed 105 out of the 133 drugs to have been previously tested against malaria, with most showing activity. For further validation, drug susceptibility assays using SYBR Green I method were done on a representative group of 10 predicted drugs, eight of which did show activity against P. falciparum 3D7 clone. Seven had IC50 values ranging from 1 μM to 50 μM. This study also suggests drug-target association and hence possible mechanisms of action of drugs that did show antiplasmodial activity. The study results validate the use of proteome-wide target similarity approach in identifying approved drugs with activity against P. falciparum and could be adapted for other pathogens.

Spermatogenesis is a complex and highly regulated process by which spermatogonial stem cells differentiate into spermatozoa. To better understand the molecular mechanisms of the process, the Cre/loxP system has been widely utilized for conditional gene knockout in mice. In this study, we generated a transgenic mouse line that expresses Cre recombinase under the control of the 2.5 kbp of the Prolactin family 3, subfamily b, member 1 (Prl3b1) gene promoter (Prl3b1-cre). Prl3b1 was initially reported to code for placental lactogen 2 (PL-2) protein in placenta along with increased expression toward the end of pregnancy. PL-2 was found to be expressed in germ cells in the testis, especially in spermatocytes. To analyze the specificity and efficiency of Cre recombinase activity in Prl3b1-cre mice, the mice were mated with reporter R26GRR mice, which express GFP ubiquitously before and tdsRed exclusively after Cre recombination. The systemic examination of Prl3b1-cre;R26GRR mice revealed that tdsRed-positive cells were detected only in the testis and epididymis. Fluorescence imaging of Prl3b1-cre;R26GRR testes suggested that Cre-mediated recombination took place in the germ cells with approximately 74% efficiency determined by in vitro fertilization. In conclusion, our results suggest that the Prl3b1-cre mice line provides a unique resource to understand testicular germ-cell development. genesis 54:389-397, 2016. © 2016 Wiley Periodicals, Inc.

Leukemias are a group of cancer types that originate from blood-forming tissues. In this disease, an abnormally large number of immature white blood cells is produced by the bone marrow. The relationship between treatments with plant-derived drugs and leukemia-associated immunophenotypes (LAIPs) of clinically isolated leukemia cells has yet to be established. The aim of the present study was to develop a preliminary clinical prognostic map for commonly expressed LAIPs in patients clinically diagnosed with leukemia, as well as to assess the potential involvement of LAIPs in the response rate to 10 natural products of plant origin. An increased expression of LAIPs, including CD4, CD14, CD33 and CD34, was considered a surrogate marker of the desired response of leukemia cells to treatment with plant-derived drugs. By contrast, the increased expression of the LAIPs, MPO and DR, was associated with poor prognostic outcomes following treatment with the plant-derived drugs. The results showed that 5 of the 10 plant-derived drugs tested induced the expression of several desirable LAIPs biomarkers. These findings clearly highlight the potential treatment efficacy of certain plant-derived drugs against leukemic cell types.