Endogenous opiates: 1989

This paper is the twelfth installment of our annual review of the research published during 1989 involving the behavioral, nonanalgesic, effects of the endogenous opiate peptides. The specific topics this year include stress; tolerance and dependence; eating; drinking; gastrointestinal and renal functions; mental illness; learning, memory, and reward; cardiovascular responses; respiration and thermoregulation; seizures and other neurological disorders; electrical-related activity; locomotor activity; sex, development, pregnancy, and aging; immunological responses; and other behavior.     

Microbial surface displaying formate dehydrogenase and its application in optical detection of formate

In the present work, NAD⁺-dependent formate dehydrogenase (FDH), encoded by fdh gene from Candida boidinii was successfully displayed on Escherichia coli cell surface using ice nucleation protein (INP) from Pseudomonas borealis DL7 as an anchoring protein. Localization of matlose binding protein (MBP)-INP-FDH fusion protein on the E. coli cell surface was characterized by SDS-PAGE and enzymatic activity assay. FDH activity was monitored through the oxidation of formate catalyzed by cell-surface-displayed FDH with its cofactor NAD⁺, and the production of NADH can be detected spectrometrically at 340 nm. After induction for 24 h in Luria-Bertani medium containing isopropyl-β-d-thiogalactopyranoside, over 80% of MBP-INP-FDH fusion protein present on the surface of E. coli cells. The cell-surface-displayed FDH showed optimal temperature of 50 °C and optimal pH of 9.0. Additionally, the cell-surface-displayed FDH retained its original enzymatic activity after incubation at 4 °C for one month with the half-life of 17 days at 40 °C and 38 h at 50 °C. The FDH activity could be inhibited to different extents by some transition metal ions and anions. Moreover, the E. coli cells expressing FDH showed different tolerance to solvents. The recombinant whole cell exhibited high formate specificity. Finally, the E. coli cell expressing FDH was used to assay formate with a wide linear range of 5–700 μM and a low limit of detection of 2 μM. It is anticipated that the genetically engineered cells may have a broad application in biosensors, biofuels and cofactor regeneration system.     

Survival Mechanisms of Entomopathogenic Nematodes

Entomopathogenic nematodes (Steinernematidae and Heterorhabditidae) are isolated from a wide variety of ecosystems ranging from sub-Arctic to arid and tropical climates. It is expected that natural populations of these nematodes will be pre-adapted to specific ecological conditions in their environments. However, little is known about the survival strategies of entomopathogenic nematodes. This review focuses on the main survival mechanisms involved in the long-term persistence of nematodes: extreme temperature tolerance (cold and heat), desiccation tolerance, osmotic tolerance, pesticide resistance and limitations of aeration. Physiological and biochemical mechanisms for survival are discussed. The review addresses the implications of the survival strategies on the establishment of exotic and indigenous steinernematids and heterorhabditids.