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.     

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.     

Risk of cancer in Finnish children living close to power lines

OBJECTIVE: To investigate the risk of cancer in children living close to overhead power lines with magnetic fields of > or = 0.01 microteslas (microT). DESIGN: Cohort study. SETTING: The whole of Finland. SUBJECTS: 68,300 boys and 66,500 girls aged 0-19 years living during 1970-89 within 500 m of overhead power lines of 110-400 kV in magnetic fields calculated to be > or = 0.01 microT. Subjects were identified by record linkages of nationwide registers. MAIN OUTCOME MEASURES: Numbers of observed cases in follow up for cancer and standardised incidence ratios for all cancers and particularly for nervous system tumours, leukaemia, and lymphoma. RESULTS: In the whole cohort 140 cases of cancer were observed (145 expected; standardised incidence ratio 0.97, 95% confidence interval 0.81 to 1.1). No statistically significant increases in all cancers and in leukaemia and lymphoma were found in children at any exposure level. A statistically significant excess of nervous system tumours was found in boys (but not in girls) who were exposed to magnetic fields of > or = 0.20 microT or cumulative exposure of > or = 0.40 microT years. CONCLUSIONS: Residentia magnetic fields of transmission power lines do not constitute a major public health problem regarding childhood cancer. The small numbers do not allow further conclusions about the risk of cancer in stronger magnetic fields.     

In vitro synthesis of molybdopterin from precursor Z using purified converting factor. Role of protein-bound sulfur in formation of the dithiolene

The pterin component of the molybdenum cofactor, termed molybdopterin, is synthesized in Escherichia coli by enzymes encoded at the chl loci. A late step in the biosynthetic pathway, the conversion of a molybdopterin intermediate, precursor Z, to molybdopterin, requires the activity of a two-subunit protein, the converting factor. Precursor Z has many of the features of molybdopterin but lacks the dithiolene function essential for molybdenum ligation. Conversion of precursor Z to molybdopterin is accomplished by transfer of sulfur to produce the dithiolene. The present study describes an in vitro system for molybdopterin biosynthesis comprised of purified precursor Z and purified converting factor. It is established that these components are sufficient to yield molybdopterin, identified by conversion to its characteristic products, Form A, Form B, and dicarboxamidomethylmolybdopterin. Under conditions of precursor excess, the formation of molybdopterin was stoichiometric with converting factor, as would be expected in the absence of a sulfur-regenerating system. The labile product of the reaction, molybdopterin, remained associated with the converting factor large subunit. These results establish that the source of sulfur for molybdopterin biosynthesis is the converting factor and suggest that in vivo a novel sulfur cycle must function to resupply sulfur to the converting factor.     

Combined Flux-Gate Instruments F-205.03, F-205.30A,and F205.38

The paper brie y describes an operation of the instruments and presents their technical characteristics.