Calcium signaling and regulation of neutrophil functions: Still a long way to go.

Neutrophils are the most abundant leukocytes in blood and disruption in their functions often results in an increased risk of serious infections and inflammatory autoimmune diseases. Following recent discoveries in their influence over disease progression, a resurgence of interest for neutrophil biology has taken place. The multitude of signaling pathways activated by the engagement of numerous types of receptors, with which neutrophils are endowed, reflects the functional complexity of these cells. It is therefore not surprising that there remains a huge lack in the understanding of molecular mechanisms underlining neutrophil functions. Moreover, studies on neutrophils are undoubtedly limited by the difficulty to efficiently edit the cell's genome. Over the past 30 years, compelling evidence has clearly highlighted that Ca2+ -signaling is governing the key processes associated with neutrophil functions. The confirmation of the role of an elevation of intracellular Ca2+ concentration has come from studies on NADPH oxidase activation and phagocytosis. In this review, we give an overview and update of our current knowledge on the role of Ca2+ mobilization in the regulation of pro-inflammatory functions of neutrophils. In particular, we stress the importance of Ca2+ in the formation of NETs and cytokine secretion in the light of newest findings. This will allow us to embrace how much further we have to go to understand the complex dynamics of Ca2+ -dependent mechanisms in order to gain more insights into the role of neutrophils in the pathogenesis of inflammatory diseases. The potential for therapeutics to regulate the neutrophil functions, such as Ca2+ influx inhibitors to prevent autoimmune and chronic inflammatory diseases, has been discussed in the last part of the review.

Immunization of flavivirus West Nile recombinant envelope domain III protein induced specific immune response and protection against West Nile virus infection.

The domain III of the West Nile virus (WNV) envelope glycoprotein (E) was shown to serve as virus attachment domain to the cellular receptor, and neutralizing Abs have been mapped to this specific domain. In this study, domain III of the WNV E protein (WNV E DIII) was expressed as a recombinant protein and its potential as a subunit vaccine candidate was evaluated in BALB/C mice. Immunization of WNV E DIII protein with oligodeoxynucleotides (CpG-DNA) adjuvant by i.p. injection was conducted over a period of 3 wk. The immunized mice generated high titer of WNV-neutralizing Abs. Murine Ab against WNV E DIII protein was also capable of neutralizing Japanese encephalitis virus. The IgG isotypes generated were predominantly IgG2a in the murine sera against the recombinant protein. Splenocyte cultures from the mice coadministrated with WNV E DIII protein and CpG secreted large amounts of IFN-gamma and IL-2 and showed proliferation of T cells in the presence of WNV E DIII protein. Overall, this study highlighted that recombinant WNV E DIII protein delivered in combination with CpG adjuvant to mice generated a Th1 immune response type against WNV and can serve as a potential vaccine to prevent WNV infection.

Hand performance assessment of ten people with rheumatoid arthritis when using a range of specified saucepans.

PURPOSE: The aim of the pilot study was to provide information about the design and use of saucepan handles to enable clinicians and designers to specify and provide products that are more appropriate for use by people with Rheumatoid Arthritis (RA). The objectives were: to evaluate aspects of new handle design for saucepans in terms of their ease of use for people with RA; document hand grip strength and configuration (grip patterns); record relevant anthropometric data to aid the development of new designs and perform an assessment of lifting techniques used in conjunction with perceived optimum handle configuration. METHOD/RESULTS: Observation and video footage show that subjects continued to use familiar, but damaging, ways of lifting the saucepan even after extensive joint protection training by occupational therapists. Grip strengths recorded using a sphygmomanometer were similar to those found by other studies. The anthropometric measurements taken from the sample group were found to be within available anthropometric surveys of able-bodied people. However, hand length within the sample group with RA was longer than the equivalent in surveys of able-bodied subjects. CONCLUSIONS: Subjects preferred the narrower handles with some surface texture to the larger and more rounded tapered handles. Large handled saucepans were found not to be viable due to the constraints of UK cooking hob sizes and existing British Standards relating to saucepan specification.

A contrastive analysis of release patterns of biochemical markers of brain damage after coronary artery bypass grafting and valve replacement and their association with the neurobehavioral outcome after cardiac surgery.

OBJECTIVE: The present study aimed at a comparative analysis of neurobiochemical markers of brain damage and the neurobehavioral outcome in patients undergoing either valve replacement (VR) or isolated coronary artery bypass surgery (CABG). In order to control for well known risk factors both samples were strictly matched according to age, sex and preoperative neuropsychological performance. METHODS: We analysed neurone-specific enolase (NSE) and protein S-100B (S-100B) concentrations in serial venous blood samples taken preoperatively and 1, 6, 20 and 30 h postoperatively in 36 patients undergoing VR (N = 18) or isolated CABG surgery (N = 18). Mini Mental State Examination (MMS) was performed preoperatively, 3 and 7 days after surgery and 0.5 years later. Neuropsychiatric assessments were based on the diagnosis of postoperative delirium according to DMS-IIIR criteria and the Brief Psychiatric and the Delirium Rating Scale. RESULTS: VR and CABG patients, respectively, showed an increase of both S-100B (exact two-tailed Wilcoxon signed ranks test: P = 0.0001) and NSE (P = 0.0001) concentrations followed by a decrease during the next 30 h. Whereas S-100B values did not differ between patients groups subjects undergoing VR surgery exhibited higher NSE values during the postoperative course. Furthermore, VR patients showed a higher decline in cognitive performance which was also detectable 0.5 years after surgery. We found a weak association between the degree of individual postoperative decline of cognitive performance and S-100B area under curve values. CONCLUSIONS: Our data indicate that--apart from patients' age and preoperative neuropsychological performance--type of surgery remains a risk factor for postoperative neurobehavioral disorders. The different vulnerability of neurobehavioral disorders might be mirrored in different postoperative release patterns of NSE. We assume that both, NSE release and neurobehavioral disorders might be caused by a higher amount of intraoperative cerebral embolic events in VR patients.

Effect of Severe Water Stress on Aspects of Crassulacean Acid Metabolism in Xerosicyos.

Xerosicyos danguyi H.Humb. (Cucurbitaceae) is a Crassulacean acid metabolism (CAM) species native to Madagascar. Previously, it was shown that when grown under good water conditions, it is a typical CAM plant, but when water stressed, it shifts to a dampened form of CAM, termed CAM-idling, in which stomata are closed day and night but with a continued, low diurnal organic acid fluctuation. We have now studied the kinetics of some metabolic features of the shift from CAM to CAM-idling under severe water stress and the recovery upon rewatering. When water is withheld, there is a steady decrease in relative water content (RWC), reaching about 50%, at which point the water potential decreases precipitously from about -2 or -3 bars to -12 bars. Abscisic acid (ABA) increases sharply at about 75% RWC. Stomata close, which limits CO2 uptake, and there is a dampened diurnal organic acid fluctuation typical of CAM-idling. Throughout an extended stress period to 50% RWC, there is no change in chlorophyll, protein, and ribulose bisphosphate carboxylase activity compared with the well-watered plants. Despite the fact that the tissue was already in CAM, the stress is accompanied by an increase in phosphoenolpyruvate carboxylase (PEPc) mRNA, extractable PEPc activity, and PEPc protein (such that the specific activity remained approximately constant) and a decrease in the apparent Km(PEP). It is not known if the changes in Km(PEP) in response to drought are related to or are separate from the increases in PEPc protein and mRNA. The changes in Km(PEP) could be in response to the decreased endogenous levels of organic acids, but evidently are not an assay artifact. The increases in PEPc protein and mRNA appear to be related to the water-stress treatment and may result from the increased concentration of ABA or the decreased levels of endogenous organic acids. When rewatered, the metabolism quickly returns to the well-watered control typical of CAM.

Liquid chromatography solvent optimization: potential pitfalls when using a black box for developing a quality separation.

ICOS and DIAMOND are two commercially available, semi-automated HPLC solvent optimization software packages. The resultant optimized chromatographic separation is dependent on a combination of the operator's objective, the capability of the software system and the appropriateness of the data input. The latter contains components that represent the match between the requirements of the algorithms used and the information content of the data on which those algorithms operated. Knowledge about the sample content, stability and potential sample-solvent interactions can have a significant effect on the quality of the optimal solvent composition that is calculated. The results generated during the optimization of the separation of a mixture of U-83,757 and a variety of related compounds illustrate the need to consider the significance of the contribution to the calculated optimal separation of each of these potential pitfalls, both individually and in combination with the mode of operation of the relevant algorithms. Our results indicate that the quality of the final result is highly dependent on the intelligence content of the data used.

The effect of instruction on dentists' motivation to manage fearful patients.

This study assesses the effect of the pre-doctoral and continuing behavioral education on dentists' efforts to treat fearful patients. A mail survey consisting of a booklet of 27 questions was sent to the 1988 and 1989 graduates of the Universities of Washington (UW), Kentucky (UK), and British Columbia (UBC). Graduates from UW and UK who received predoctoral instruction and some UBC graduates who received continuing education on the behavioral approach to caring for fearful patients were compared with UBC graduates who received only a primarily pharmacological oriented education. The overall response rate was 80.4 percent (164/204). Results indicate the average proportion of fearful patients seen was greater for those receiving behavioral instruction. Dentists who received behavioral instruction also reported greater effort addressing fear during patient's initial contact with the practice; however, they perceived the costs of the behavioral approach to be greater. No differences were found between groups for the remaining effort measures.

Use of high-performance liquid chromatographic peak deconvolution and peak labelling to identify antiparasitic components in plant extracts.

Artemisia absynthium L. is a commonly used medicinal plant for parasitic diseases all over the world. By means of high-performance liquid chromatography with diode-array detection and the PU6100 solvent optimization system, two sesquiterpene lactones, alpha-santonin and ketopelenolid-A, were tentatively identified in methanolic extracts of this plant. alpha-Santonin is a well known antiparasitic compound and could be one of the active principles of this plant species. Reconstructed spectra are potentially useful in scanning a complex chromatogram for pharmacologically active compounds.

Oral neurofibrosarcoma associated with neurofibromatosis type I.

One of the most feared complications of neurofibromatosis type I (NF-I) is development of cancer, which is estimated to occur in about 5% of cases. The most common associated malignancy is the neurofibrosarcoma (NFS). HOwever, oral NFS in association with NF-I has rarely been reported. We report two cases of oral NFS arising in patients with NF-I. Both patients died of their tumors. Oral NFS arising in association with NF-I appears to have an extremely poor prognosis, as do these tumors at other sites of the body.

Ecophysiological comportment of the tropical CAM-tree Clusia in the field: II. Modes of photosynthesis in trees and seedlings.

Measurements were performed on leaves of Clusia rosea Jacq. trees in the moist central mountains (330 to 365 m above sea level) and at the dry south coast of St John Island (US Virgin Islands, Lesser Antilles). Seedlings of C. rosea were also studied in the central hills. During the study period (March 1989) all trees showed crassulacean acid metabolism (CAM), in which net CO2 uptake extended for a remarkably long time in the morning (phase II of CAM: until about 11 to 12 h) and contributed about 1/3 of total net CO2 -uptake. During the night (phase I of CAM) malic acid and citric acid were accumulated concurrently at a molar ratio of malic: citric acid of about 1.6. Internal recycling of respiratory CO2 was 20% of total CO2 fixed during the night. Water-use-efficiency (mol CO2 taken up: mol H2 O transpired) was 0.014 to 0.022. The pH of leaf-cell sap at the end of the dark period was 2.85. This would still allow an H+ -ATPase at the tonoplast to transport 2H+ into the vacuole per ATP hydrolysed when operating near thermodynamic equilibrium. Free sugars, glucose and fructose, and starch were used as precursors for the CO2 -acceptor phosphoenolpyruvate during the dark period; contributions of the two hexoses were about equal and together four-times that of starch. Xylem tensions showed increases of up to 8 bar during day-time. Leaf-sap osmotic pressures did not change significantly; the trend was a small decline during day-time. Among the seedlings, three different modes of photosynthesis were encountered, namely C3 -photosynthesis in terrestrial and in epiphytic seedlings, continuous stomatal opening and CO2 -uptake day and night in epiphytic seedlings, and CAM in seedlings growing in the tanks of Aechmea lingulata (L.) Baker.

Ecophysiological comportment of the tropical CAM-tree Clusia in the field: I. Growth of Clusia rosea Jacq. on St John, US Virgin Islands, Lesser Antilles.

Clusia rosea Jacq. is abundant in the moist parts of the Caribbean island of St John (US Virgin Islands, Lesser Antilles) but relatively rare along the dry south coast. Three types of seedlings were encountered, terrestrial seedlings, seedlings growing as humus-epiphytes on other trees, and seedlings growing inside the tanks of the bromeliad Aechmea lingulata (L.) Baker. Free-living trees grow from terrestrial seedlings or from epiphytic seedlings strangling and shading their host trees. Leaf-Na+ levels were always low (1-4 mequiv I-1 tissue water); trees close to the shore were not affected by salinity. In leaves of mature C. rosea trees, levels of Ca2+ , Mg2+ and K+ were about 60-90, 40-50, 45-55 mequiv I-1 tissue water, respectively. Epiphytic seedlings tended to contain lower levels of these inorganic cations than seedlings growing terrestrially or in the tanks of Ae. lingulata. Epiphytic seedlings contained significantly less nitrogen than terrestrial seedlings. In the leaves of mature trees N-levels were independent of altitude and location on the island, but shaded leaves had significantly higher N-levels than exposed leaves. Light compensation point of photosynthesis in epiphytic seedlings performing C3 -photosynthesis was 17-5 (µmol photons m-2 s-1 ), photosynthesis was saturated at about 300µmol photons m-2 s-1 showing a maximum rate of CO2 -uptake of 2-3 µmol m-2 s-1 .

Clinical experience with ciprofloxacin.

In a multicenter study of 30 patients treated with ciprofloxacin (mean daily dosage, 1000 mg per day; mean duration of treatment, 10.6 days) for a variety of infections, 14 were microbiologically proven. Of these, bacteriologic cure and/or improvement resulted in 92.3% of cases. For all 30 infections clinical cure and/or improvement resulted in 88.9% of cases. A total of ten infections were classified as chronic. Overall there were 5/30 (16%) adverse reactions (ADRs), however, only two were definitely related to ciprofloxacin therapy. For three ADRs the relationship to therapy was uncertain. Therapy with ciprofloxacin was discontinued in one (3.3%) patient because of adverse effects. Four patients elected to continue ciprofloxacin therapy despite mild side effects.

Rapid automated high-performance liquid chromatographic assay for ercalcidiol and calcidiol (25-hydroxyvitamins D2 and D3) using trans-calcidiol as an ultraviolet-absorbing internal standard.

A system is described using high-performance liquid chromatography to separate and quantify, by spectrophotometry in a simple one-stage procedure, ercalcidiol (25-hydroxyvitamin D2) and calcidiol (25-hydroxyvitamin D3). The novel feature of the method is the employment of an ultraviolet-absorbing internal standard to monitor recovery. This has the advantage of permitting total automation of the quantification by eliminating the need for radioactivity counting. The method gives results that compare well with those obtained in other systems and has particular application in clinical studies where rapid separate determination of ercalcidiol and calcidiol is required.

Photosynthesis in epiphytic and rooted Clusia rosea Jacq.

Clusia rosea Jacq. is a hemiepiphyte having Crassulacean Acid Metabolism (CAM). In its natural habitat Clusia begins its life cycle as an epiphyte and eventually becomes a rooted tree. These two stages of the life cycle of Clusia represent markedly different water regimes. Our CO2 exchange, stomatal conductance, titratable acidity, and stable carbon isotope ratio measurements indicate that Clusia has a flexible photosynthetic mode, where CO2 is fixed mostly via CAM during its epiphytic stage, when water availability is low, and via both CAM and C3 during its rooted stage.