Bt Eggplant Project in Bangladesh: History, Present Status, and Future Direction.

The purpose of this article is to provide information on the history, accomplishments, and future direction of the Bt brinjal (eggplant) program in Bangladesh, formerly under the Agricultural Biotechnology Support Project II, now the South Asia Eggplant Improvement Partnership (SAEIP). The India-based Maharashtra Hybrid Seed Company (Mahyco) developed an eggplant expressing Cry1Ac (EE-1) for control of the eggplant fruit and shoot borer (EFSB). In a partnership among Mahyco, USAID, Sathguru Management Consultants and Cornell University EE-1 was provided to the Bangladesh Agricultural Research Institute (BARI) who bred it into local varieties. After regulatory approval, four varieties were distributed to 20 farmers who harvested Bt brinjal in 2014. Adoption in subsequent years has increased rapidly so that, in 2018, 27,012 farmers used this technology. This article provides background information on the process leading up to current adoption levels, the level of control of EFSB achieved and the economic benefits of Bt brinjal. Efforts on stewardship, farmer training and communication are discussed. In order to ensure the long-term future of the partnership, we discuss the need to enhance involvement of the private sector in the production and stewardship of Bt eggplant. Bt brinjal is the first genetically engineered crop to be commercially released in Bangladesh, and other GE crops are in the pipeline. Hence, success of the Bt brinjal partnership is likely to affect the future of other GE crops in Bangladesh, as well as other parts of the world where biotechnology is needed for food security and environmental safety.

Randomized clinical trial of a modified Seldinger technique for open central venous cannulation for implantable access devices.

INTRODUCTION: Totally implantable access ports (TIAPs) are often used for patients who need permanent venous access. The primary success rate using direct open insertion is about 80 per cent, so rescue strategies are needed. This study compared the primary success rates of standard open insertion and a modified Seldinger technique. METHODS: This randomized trial recruited 164 patients scheduled for primary implantation of a TIAP and compared two interventions. The primary endpoint was the success rate of the implantation technique. RESULTS: The primary success rates were similar: 66 (80 per cent) of 82 patients who had standard open insertion versus 69 (84 per cent) of 82 patients undergoing the modified Seldinger method (P = 0.686). A logistic mixed regression analysis including treatment group, age, Karnofsky index, body mass index and surgeon's experience showed no advantage for the Seldinger method: odds ratio 1.30 (95 per cent confidence interval 0.62 to 2.70). TIAPs were eventually implanted successfully in 163 (99.4 per cent) of 164 patients. In 11 patients randomized to standard surgery, the Seldinger method was a successful rescue strategy. CONCLUSION: The primary success rate was similar for both open insertion methods. The modified Seldinger method is useful if standard open insertion fails. REGISTRATION NUMBER: ISRCTN 52368201 (http://www.controlled-trials.com).

Efficient mobilization of peripheral blood stem cells following CAD chemotherapy and a single dose of pegylated G-CSF in patients with multiple myeloma.

High-dose chemotherapy followed by autologous blood stem cell transplantation is the standard treatment for myeloma patients. In this study, CAD (cyclophosphamide, adriamycin, dexamethasone) chemotherapy and a single dose of pegfilgrastim (12 mg) was highly effective in mobilizing peripheral blood stem cells (PBSCs) for subsequent transplantation, with 88% of patients (n = 26) achieving the CD34+ cell harvest target of > or = 7.50 x 10(6) CD34+ cells/kg body weight, following a median of two apheresis procedures (range 1-4) and with first apheresis performed at a median day 13 after CAD application (range 10-20). Patients treated with pegfilgrastim showed a reduced time to first apheresis procedure from mobilization compared with filgrastim-mobilized historical matched controls (n = 52, P = 0.015). The pegfilgrastim mobilization regimen allowed for transplantation of a median of 3.58 x 10(6) CD34+ cells/kg body weight while leaving sufficient stored cells for a second high-dose regimen and back-ups in most patients. Engraftment following transplantation was comparable to filgrastim, with a median time of 14 days to leucocyte > or =1.0 x 10(9)/l (range 10-21) and 11 days to platelets > or = 20 x 10(9)/l (range 0-15). The results of this study thus provide further support for the clinical utility of pegfilgrastim for the mobilization of PBSC following chemotherapy in cancer patients scheduled for transplantation.

Characterization of digestive proteolytic activity in Lygus hesperus Knight (Hemiptera: Miridae).

The tarnished plant bug, Lygus hesperus Knight, is a pest that causes considerable economic losses to vegetables, cotton, canola, and alfalfa. Detailed knowledge of its digestive physiology will provide new opportunities for a sustainable pest management approach to control this insect. Little is known about the different protease class contributions to the overall digestion of a specific protein. To this end, the proteolytic activities in female adult L. hesperus salivary gland and midgut homogenates were quantified over a range of pH's and time points, and the contribution of different classes of proteases to the degradation of FITC-casein was determined. In the salivary gland, serine proteases were the predominant class responsible for caseinolytic activity, with the rate of activity increasing with increasing pH. In contrast, both aspartic and serine proteases contributed to caseinolytic activity in the midgut. Aspartic protease activity predominated at pH 5.0 and occurred immediately after incubation, whereas serine protease activity predominated at pH 7.5 after a 9h delay and was resistant to aprotinin. The salivary serine proteases were distinctly different from midgut serine proteases, based on the tissue-specific differential susceptibility to aprotinin and differing pH optima. Collectively, the caseinolytic activities complement one another, expanding the location and pH range over which digestion can occur.

Uptake, flow, and digestion of casein and green fluorescent protein in the digestive system of Lygus hesperus Knight.

Selected compounds were used to study physiological processes associated with digestion in the western tarnished plant bug, Lygus hesperus Knight. Durations of passage and rates of absorption, digestion, and excretion were determined for a digestible protein (casein), a non-digestible protein (green fluorescent protein, GFP), and a non-digestible carbohydrate (dextran). Dextran was used as a control to monitor the non-absorptive flow rate of ingesta through the digestive system. Fluorescent tracking of FITC-conjugates of casein and dextran, as well as immunoblotting and immunofluorescent staining of casein and GFP, were used to monitor the degradation (in vitro) and ingestion, digestion, and distribution (in vivo) of the respective compounds. Under our experimental conditions, L. hesperus took discrete meals, feeding and excreting at 2-3 h intervals. Rate of food passage was variable. FITC-dextran was found in the fecal material of most insects by 6-8 h after treatment initiation; by 12 h, 95% of ingested FITC-dextran was recovered from all insects. FITC-casein was digested extensively in in vitro homogenates of gut, hemolymph, and salivary gland. In vivo, FITC-casein was ingested and partially absorbed as a holoprotein into the hemolymph. Ingested FITC-casein was partially degraded in the gut and hemolymph within 2 h of ingestion, and no holoprotein was found after 12 h. In contrast, there was no detectable degradation of GFP in hemolymph, gut, and salivary gland homogenates after 24 h of incubation. Ingested GFP was not degraded in gut or hemolymph up to 8 h after treatment initiation, but did transfer to the hemolymph as a holoprotein. Analysis of immunohistological images confirmed that GFP bound to gut epithelial cell brush-border membranes. However, the mechanism by which GFP and casein pass as holoproteins into the hemolymph remains unknown.

Cystatin C--an accurate marker of glomerular filtration rate after renal transplantation?

The performance of serum cystatin C as a screening marker of reduced creatinine clearance in renal transplantation was evaluated and compared to serum creatinine. In addition we studied whether cystatin C accurately reflects creatinine clearance over the entire range of transplant function. Serum cystatin C, serum creatinine, and creatinine clearance were measured in 110 adult renal transplant recipients. Cystatin C detected reduced creatinine clearance with the high sensitivity of 95%. Serum cystatin C and serum creatinine did not differ regarding 90 and 95% sensitivity, derived from the receiver-operating characteristics plot. We demonstrated a strong correlation and linear association between 1/cystatin C and creatinine clearance over the entire range of transplant function, equivalent to that of 1/creatinine. In summary, serum cystatin C accurately reflects creatinine clearance over the entire range of transplant function and is as efficacious as serum creatinine to detect reduced creatinine clearance in renal transplant recipients.

An insecticidal N-acetylglucosamine-specific lectin gene from Griffonia simplicifolia (Leguminosae).

Griffonia simplicifolia II, an N-acetylglucosamine-specific legume lectin, has insecticidal activity when fed to the cowpea weevil, Callosobruchus maculatus (F.). A cDNA clone encoding G. simplicifolia II was isolated from a leaf cDNA library, sequenced, and expressed in a bacterial expression system. The recombinant protein exhibited N-acetylglucosamine-binding and insecticidal activity against cowpea weevil, indicating that glycosylation and multimeric structure are not required for these properties. These results support the hypothesis that genes of the legume lectin gene family encode proteins that function in plant defense against herbivores.

Alpha-amylase inhibitor, not phytohemagglutinin, explains resistance of common bean seeds to cowpea weevil.

There are claims that phytohemagglutinin (PHA), the lectin of common bean, Phaseolus vulgaris, is toxic when fed to the cowpea weevil, Callosobruchus maculatus, and that PHA serves as the chemical defense against this seed-feeding bruchid beetle (DH Janzen, HB Juster, IE Liener [1976] Science 192: 795-796; AMR Gatehouse, FM Dewey, J Dove, KA Fenton, A Pusztai [1984] J Sci Food Agric 35: 373-380). However, our studies indicate that neither PHA nor its isolectins have detrimental effects when fed to the cowpea weevil. To explain these contradictory results we characterized the commercial lectin source used by A. M. R. Gatehouse, F. M. Dewey, J. Dove, K. A. Fenton, A. Pusztai (1984, J Sci Food Agric 35: 373-380). We demonstrate here that the toxic effects of PHA to cowpea weevil are due to an alpha-amylase inhibitor contaminant in the commercial preparation.

Biochemical investigations of antibiosis material in leaf exudate of wildNicotiana species and interspecific hybrids.

A model system involving severalNicotiana species containing novel nicotine alkaloids was used to study heritability and expression of alkaloid production in leaf trichomes. The three species that comprise the section Repandae (N. repanda, N. stocktonii, andN. nesophila) were hybridized with eitherN. tabacum orN. sylvestris (neither of which producesN-acylnornicotine). The progeny of the hybrid with sylvestris produced theN-acylnornicotines at a level found in the Repandae parent.Nicotiana repanda was crossed toN. tabacum, and the F2 progeny produced the alkaloid at the same level as the original Repandae parent. Inheritance of the ability to acylate nornicotine in Repandae species is inherited in hybrids in a dominant manner These and other data obtained suggest that theN-acyltransferase that acylates nornicotine in Repandae species inherited in hybrids is in a dominant manner and that the regulatory sequence(s) for the gene is expressed in leaf trichomes when the gene is in a foreignNicotiana background.

Identification of tobacco hornworm antibiosis factor from cuticulae of repandae section ofNicotiana species.

Cuticular components of the green leaves of the Repandae section of theNicotiana species contain compounds that have been shown to be toxic to larvae of the tobacco hornworm larvae,Manduca sexta. The surface constituents of leaves of greenhouse-grownN. repanda, N. stocktonii, andN. nesophila were extracted with methylene chloride in order to isolate the active compounds. Solvent partitioning and gel chromatography was used to isolate a series of hydroxyacylnornicotines (HOAcylNN). The major component was identified asN'-(3-hydroxy-12-methyltridecanoyl)nornicotine. A number of minor 3-hydroxyacylnornicotines, with the acyl group containing C13-C15, were also identified. The HOAcylNN mixture in ethanol was topically applied to first-instar tobacco hornworm larvae at rates of 10, 50, 100, and 200 µg. Mortalities after 48 hr were 33, 44, 78, and 100% respectively.