Dye affinity chromatography for fast and simple purification of fucoidan from marine brown algae

Fucoidan is a sulfated polysaccharide with promising pharmacological applications. Due to its medicinal properties, there is a demand for a separation technique that yields a high purification grade. Here, we present a novel purification tool for recovering fucoidan from the marine brown macroalgae Fucus vesiculosus. The developed method is based on amino‐derivatized Sepabeads^® EC‐EA. The beads were modified with toluidine blue (TB), a thiazine derivative, to exploit the strong donor acceptor interactions between the cationic dye and the anionic polysaccharide. The adsorption kinetics and the binding capacity of the resin were analyzed. A Sips model was used to approximate the adsorption isotherm, resulting in a maximum capacity of 127.7 mg fucoidan per g adsorbent. Investigation of the effect of adsorption step's pH on purity and chemical structure was performed by TB and Fourier transform infra‐red spectroscopy assays. Results showed that adsorption at pH 1 and 6 had negligible effects on fucoidan's chemical structure. However, purity was actually improved by 1.55‐ and 1.69‐fold at pH 1 and 6, respectively, with an average yield of 5 g/100 g dried algae powder. In contrast, only a 1.46‐fold increment was observed in fucoidan purified by the traditional method at pH 2, with a yield of 7.5 g/100 g dried algae powder. Furthermore, fucoidan purified by this method at pH 6 complies with, or even exceeds the quality of the commercially available (≥95% pure) fucoidan (Sigma‐Aldrich^®) with respect to molecular weight and sulfur content. Therefore, dye affinity chromatography provides more advantages than the classically used techniques for fucoidan purification.     

Bio-active composts from rice straw enriched with rock phosphate and their effect on the phosphorous nutrition and microbial community in rhizosphere of cowpea

Composts were produced from rice straw enriched with rock phosphate and inoculated with Aspergillus niger, Trichoderma viride and/or farmyard manure (FYM). The resulting composts were evaluated as organic phosphate fertilizers for cowpea plants in pot experiments. The results showed that the maximum amount of soluble phosphorous (1000 ppm) was produced in composts inoculated with A. niger+T. viride with or without FYM. Any of the produced composts was much better than superphosphate fertilizer in providing the growing cowpea plants with phosphorous. Fertilization of the cowpea plants with the compost inoculated with FYM+A. niger+T. viride resulted in maximum amount of phosphorous uptake (295 ppm). The highest phosphate dissolving fungi numbers in rhizosphere soils of cowpea plants were obtained after fertilization with composts which received A. niger and T. viride treatments, while the highest phosphate dissolving bacterial numbers were found after fertilization with composts which received FYM treatments.     

Direct capture of plasmid DNA from non-clarified bacterial lysate using polycation-grafted monoliths

Monolith columns from macroporous polyacrylamide gel were grafted with polycations, poly(N,N-dimethylaminoethyl methacrylate) (polyDMAEMA), (2-(methacryloyloxy)ethyl)-trimethyl ammonium chloride (polyMETA) and partially quaternized polyDMAEMA prepared via treating polyDMAEMA-grafted columns with propylbromide. The polymer grafting degrees varied between 34 and 110%. The polycation-grafted monolithic columns are able to capture plasmid DNA directly from alkaline lysate of Escherichia coli cells. Due to the large pore size in macroporous monoliths the particulate material present in non-clarified feeds did not block the columns. The captured plasmid DNA was eluted with 1M NaCl as particulate-free preparation with significantly reduced content of protein and RNA as compared to the applied lysate.     

Programmed reduction of ABC transporter activity in sea urchin germline progenitors

ATP-binding cassette (ABC) transporters protect embryos and stem cells from mutagens and pump morphogens that control cell fate and migration. In this study, we measured dynamics of ABC transporter activity during formation of sea urchin embryonic cells necessary for the production of gametes, termed the small micromeres. Unexpectedly, we found small micromeres accumulate 2.32 times more of the ABC transporter substrates calcein-AM, CellTrace RedOrange, BoDipy-verapamil and BoDipy-vinblastine, than any other cell in the embryo, indicating a reduction in multidrug efflux activity. The reduction in small micromere ABC transporter activity is mediated by a pulse of endocytosis occurring 20-60 minutes after the appearance of the micromeres--the precursors of the small micromeres. Treating embryos with phenylarsine oxide, an inhibitor of endocytosis, prevents the reduction of transporter activity. Tetramethylrhodamine dextran and cholera toxin B uptake experiments indicate that micromeres have higher rates of bulk and raft-associated membrane endocytosis during the window of transporter downregulation. We hypothesized that this loss of efflux transport could be required for the detection of developmental signaling molecules such as germ cell chemoattractants. Consistent with this hypothesis, we found that the inhibition of ABCB and ABCC-types of efflux transporters disrupts the ordered distribution of small micromeres to the left and right coelomic pouches. These results point to tradeoffs between signaling and the protective functions of the transporters.     

Genetic differentiation across a behavioural boundary in a primitively eusocial bee, <Emphasis Type="Italic">Halictus poeyi</Emphasis> Lepeletier (Hymenoptera,Halictidae)

Summary. In Florida, the primitively eusocial bee, Halictus poeyi Lepeletier exhibits two different colony cycles: populations in northern and central Florida are univoltine with an annually brooded colony cycle, while populations in southern Florida, including the Florida Keys, are multivoltine with a continuously brooded colony cycle. In this study, we test the hypothesis of reduced gene flow between the colony types by comparing levels of genetic differentiation among populations having similar colony phenologies with that among populations having different colony phenologies. We use allozyme markers to estimate F_ST, a robust measure of genetic differentiation. We found that genetic differentiation is not significantly higher between populations having different colony phenologies than between populations having similar colony phenologies. Environmental conditions thus may play an important role in influencing the expression of the alternative colony types of H. poeyi populations in Florida.     

Fusarithioamide B,a new benzamide derivative from the endophytic fungus Fusarium chlamydosporium with potent cytotoxic and antimicrobial activities

Fusarithioamide B (6), a new aminobenzamide derivative with unprecedented carbon skeleton and five known metabolites: stigmast-4-ene-3-one (1), stigmasta-4,6,8(14),22-tetraen-3-one (2), p-hydroxyacetophenone (3), tyrosol (4), and fusarithioamide A (5) were separated from Fusarium chlamydosporium EtOAc extract isolated from Anvillea garcinii (Burm.f.) DC. leaves (Asteraceae). The structure elucidation and complete assignment of the isolated metabolites were performed mainly by the aid of various NMR and MS data. Fusarithioamide B (6) has been assessed for antibacterial and antifungal activities towards various microbial strains by disc diffusion assay. It exhibited selective antifungal activity towards C. albicans (MIC 1.9?µg/ml and IZD 14.5?mm), comparing to clotrimazole (MIC 2.8?µg/ml and IZD 17.9?mm). Also, it possessed high antibacterial potential towards E. coli, B. cereus, and S. aureus compared to ciprofloxacin. Furthermore, 6 was tested for the in vitro cytotoxic effect against KB, HCT-116, BT-549, MCF-7, SKOV-3, and SK-MEL cell lines. It had selective and potent effect towards BT-549, MCF-7, SKOV-3, and HCT-116 cell lines with IC₅₀s 0.09, 0.21, 1.23, and 0.59?μM, respectively compared to doxorubicin (IC₅₀s 0.046, 0.05, 0.321, and 0.24?μM, respectively). Fusarithioamide B may provide a lead molecule for future developing of antitumor and antimicrobial agents.     

Age patterns of Cryptosporidium species and Giardia duodenalis in dairy calves in Egypt

Little is known of the occurrence and age patterns of species/genotypes and subtypes of Cryptosporidium spp. and Giardia duodenalis in calves in Egypt. In this study, 248 fecal specimens were collected from dairy calves aged 1?day to 6?months on eight farms in three provinces during March 2015 to April 2016. Cryptosporidium spp. were detected and genotyped by using PCR-RFLP analysis of the small subunit rRNA (SSU rRNA) gene, while G. duodenalis was detected and genotyped by using PCR and sequence analyses of the triose phosphate isomerase (tpi), glutamate dehydrogenase (gdh) and β-giardin (bg) genes. The overall infection rates of Cryptosporidium spp. and G. duodenalis were 9.7 and 13.3%, respectively. The highest Cryptosporidium infection rate (26.7%) was in calves of age?≤?1?month while the highest G. duodenalis infection rate (44.4%) was in calves of 2?months. Three Cryptosporidium spp. were identified, including C. parvum (n?=?16), C. bovis (n?=?5) and C. ryanae (n?=?3), with the former being almost exclusively found in calves of ≤3?months of age and the latter two being only found in calves of over 3?months. Subtyping of C. parvum by PCR-sequence analysis of the 60?kDa glycoprotein gene identified subtypes IIaA15G1R1 (n?=?15) and IIaA15G2R1 (n?=?1). The G. duodenalis identified included both assemblages E (n?=?32) and A (n?=?1), with the latter belonging to the anthroponotic subtype A2. These data provide new insights into the genetic diversity and age patterns of Cryptosporidium spp. and G. duodenalis in calves in Egypt.     

Reverse Problem Formulation for Integrating Process Discharges with Watersheds and Drainage Systems

This work introduces a new approach to integrating the discharges of industrial processes with macroscopic watershed systems. The key concept is that environmental quality models (such as material flow analysis) can be inverted and included in an optimization formulation that seeks to determine the maximum allowable target for the process discharges while meeting the overall environmental requirements of the watershed. Because of its holistic nature, this approach simultaneously considers the effects of the inputs and outputs to the watershed (e.g., agricultural, residential, wastewater treatment plants, industrial, and so on) and the various physical, chemical, and biological phenomena occurring within the watershed. An optimization formulation is developed to systematically represent the reverse problem formulation. To illustrate the effectiveness of this approach, a case study is solved to manage phosphorus in Bahr El‐Baqar drainage system leading to Lake Manzala in Egypt. The key environmental and economic aspects are addressed and used to screen plant location and discharges.