The resistance of cowpea seeds to bruchid beetles is not related to levels of cysteine proteinase inhibitors

A cDNA encoding a cysteine proteinase inhibitor was isolated from a cDNA library prepared from developing seeds of an insect-resistant line of cowpea. The sequence of the encoded protein was homologous with those of other plant cysteine endoproteinase inhibitors, and with Type 2 cystatins from animals. Southern blot analyses indicated that small gene families were present in both resistant and susceptible lines of cowpea, while northern blot analyses showed similar levels of expression. It is concluded that the levels of expression of the inhibitor do not account for the differences in insect resistance of the two lines.     

Cisplatin resistance and regulation of DNA repair in cAMP-dependent protein kinase mutants

Drug resistance in cancer poses a major problem to the success of chemotherapy. Increased resistance to the DNA-damaging chemotherapeutic drug cisplatin may be associated with a variety of factors including decreased drug accumulation, increased intracellular levels of thiols, and increased DNA repair. We have found that mutants of the Chinese hamster ovary (CHO) and the mouse adrenocortical carcinoma Y1 cells harboring a defective regulatory subunit (RI) of the cAMP-dependent protein kinase (PKA) exhibited increased resistance to cisplatin. These mutants are cross-resistant to other DNA-damaging chemotherapeutic agents, including bleomycin and melphalan. In addition, wild-type CHO cells transfected with and overexpressing the yeast phosphodiesterase gene or a dominant mutant Rl alpha subunit gene also displayed similar increased resistance to cisplatin. However, mutants with altered catalytic (C) subunits showed a sensitivity to cisplatin similar to the wild-type cells. Further analysis by gel shift assay using cisplatin-damaged DNA as probes and nuclear extracts derived from the Rl subunit mutants showed increased binding of nuclear factor(s) to the damaged DNA. In addition, a host cell reactivation assay of DNA repair, using a cisplatin-damaged reporter plasmid, detected enhanced capacity for repair of DNA lesions in the PKA mutants. These results suggest that DNA repair may be increased in the PKA mutants. We speculate that functional inactivation of PKA may result in increased DNA repair and the acquisition of resistance to DNA-damaging anticancer drugs in cancer.     

The ice-binding site of Atlantic herring antifreeze protein corresponds to the carbohydrate-binding site of C-type lectins

The type II antifreeze proteins (AFPs) of smelt and Atlantic herring are homologous to the carbohydrate-recognition domains (CRDs) of Ca2+-dependent (C-type) animal lectins and, like these lectins, acquire a stable and active structure upon binding Ca2+ ions. In the C-type lectin CRD, the carbohydrate-binding site is located at a Ca2+-binding site. Site-directed mutagenesis was used to test the hypothesis that the ice-binding site of the type II AFP corresponds to the carbohydrate-binding site of the lectins. To disrupt this site in the herring AFP without perturbing the Ca2+-dependent protein fold, a double mutant was constructed that changed the Ca2+- and carbohydrate-binding motif from the galactose-type of wild-type AFP containing the sequence Gln-Pro-Asp to a mannose-type that has the sequence Glu-Pro-Asn and is also known to bind Ca2+. The mutant AFP exhibited proper Ca2+ binding, folding, and stability as demonstrated by ruthenium red staining, proteolysis protection assays, and CD spectroscopy. However, it showed no antifreeze activity (thermal hysteresis) and did not alter ice crystal morphology to form bipyramidal crystals as does the active wild-type AFP. These results demonstrate that the ice-binding site of the herring type II AFP corresponds to the carbohydrate-binding site of the C-type lectin CRDs and further suggest that this ice-binding function evolved from the carbohydrate-binding site of a preexisting C-type lectin.     

The surface renewal model of wall turbulence for boundary layer flow with transpiration

The object of this paper is to develop and evaluate the basic surface renewal modeling approach for transpired turbulent boundary layer flows. Using a simple form of the surface renewal model in conjunction with the standard mixing length representation of the turbulent core, calculations are established for the dimensionless burst frequency s⁺, and distributions in velocity u⁺ within the inner region as a function of transpiration rate v⁺ and pressure gradient P⁺.     

Dual disabilities: when a stroke patient fractures a hip

The incidence of both hip fracture and stroke increases with age. With the increasing age of the United States population, it is expected that the orthopaedic nurse will be challenged to care for clients with the dual disabilities of hip fracture and stroke. This article discusses incidence of these disabilities, describes the pathophysiology of stroke, and uses a case study to outline specific nursing interventions for the patient with hip fracture and stroke.     

Study of serum antibodies of different classes as a methods of defining the genesis of transitory salmonella carrier state]

An 18 month Thoroughbred gelding was diagnosed on cardiac catheterization as having an interventricular septal defect. Right side cardiac blood pressures were within the normal range and confirmation of the diagnosis came from changes in the blood pO2 and pCO2 between the right atrium and right ventricle. The significance of these values is discussed.     

Validation of a new blood-mimicking fluid for use in Doppler flow test objects

PM28A is a major intrinsic protein of the spinach leaf plasma membrane and the major phosphoprotein. Phosphorylation of PM28A is dependent in vivo on the apoplastic water potential and in vitro on submicromolar concentrations of Ca2+. Here, we demonstrate that PM28A is an aquaporin and that its water channel activity is regulated by phosphorylation. Wild-type and mutant forms of PM28A, in which putative phosphorylation sites had been knocked out, were expressed in Xenopus oocytes, and the resulting increase in osmotic water permeability was measured in the presence or absence of an inhibitor of protein kinases (K252a) or of an inhibitor of protein phosphatases (okadaic acid). The results indicate that the water channel activity of PM28A is regulated by phosphorylation of two serine residues, Ser-115 in the first cytoplasmic loop and Ser-274 in the C-terminal region. Labeling of spinach leaves with 32P-orthophosphate and subsequent sequencing of PM28A-derived peptides demonstrated that Ser-274 is phosphorylated in vivo, whereas phosphorylation of Ser-115, a residue conserved among all plant plasma membrane aquaporins, could not be demonstrated. This identifies Ser-274 of PM28A as the amino acid residue being phosphorylated in vivo in response to increasing apoplastic water potential and dephosphorylated in response to decreasing water potential. Taken together, our results suggest an active role for PM28A in maintaining cellular water balance.     

Toxic effect of systemic administration of low doses of the plasticizer di-(2-ethyl hexyl) phthalate [DEHP] in rats

A spreadsheet model of a circle breathing system and a 70-kg anaesthetised 'standard man' has been used to simulate the first 20 min of low-flow anaesthesia with halothane, enflurane, isoflurane, sevoflurane and desflurane in oxygen. It is shown that, with the fresh-gas flow set initially equal to the total ventilation and the fresh-gas partial pressure to 3 MAC, the end-expired partial pressure can be raised to 1 MAC in 1 min with desflurane and sevoflurane, 1.5 min with isoflurane, 2.5 min with enflurane and 4 min with halothane. Sequences of lower fresh-gas flow and partial pressure settings are given for then maintaining 1 MAC end-expired partial pressure, with a minimum usage of anaesthetic, e.g. 13 ml of liquid desflurane in 20 min (of which only 33% is taken up by the patient) if the minimum acceptable flow is 11.min-1, or 8 ml (with 57% in the patient) if the minimum is 250 ml.min-1.