Blood trauma in plastic haemodialysis cannulae

To investigate the haemolysis in haemodialysis cannulae, an in-vitro set up is built, using a unipuncture dialysis system. This system is connected to a bag with fresh calf's blood, by the cannula under test, mounted in a large bloodline (5 mm diameter). The blood characteristics are kept constant by means of a bicarbonate dialysate in the dialyser. During a 6 h period, haematological parameters are regularly samples. Flow through the cannulae is recorded, which is about 500 mL/min. Four different cannulae are tested and compared to the results obtained without any cannula in the circuit. In all cases a linear increase in plasma free haemoglobin levels is found after 6 h. The cannulae can be ranked from 8F catheter over 13G, 14G to 16G cannula, the latter producing the highest degree of haemolysis. When using plastic cannulae at high blood flows, their haemolytic effect may not be neglected.     

Distribution of protein phosphatases type 1 and 2A in RINm5F cells

In the insulin producing cell line RINm5F distribution of serine/threonine specific protein phosphatases type 1 (PP1) and 2A (PP2A) was studied. Using different agents which inhibit or stimulate PP1 and PP2A we found that in membrane and nuclear fractions phosphatase activity was inhibited by okadaic acid (OA), protamine, heparin, and inhibitor-2 in a concentration-dependent manner. C2-ceramide had no effect. In the cytosolic fraction the inhibitory effect of okadaic acid was tenfold higher. Protamine stimulated phosphatase activity at low concentrations and became inhibitory at higher concentrations. Inhibitor-2 and heparin caused a decrease in phosphatase activity whereas C2-ceramide led to a slight activation. The data suggest that in membrane and nuclear fractions of RINmSF cells predominantly PP1 is present, whereas in the cytosol PP1 as well as PP2A can be detected.     

Vigabatrin-induced lesions in the rat brain demonstrated by quantitative magnetic resonance imaging

Previous research has indicated that individual foods or beverages are ingested independently and do not produce adjustments to the intake of other constituents in the diet (de Castro, 1993; Wilson, 1991). In order to eliminate time of day as a potential contaminant, the present study investigated the accommodation of foods and beverages into the amount ingested at large evening meals only. Adults (n = 601) were paid to maintain detailed diaries of the timings, quantities and preparation techniques of everything they ingested for seven consecutive days. With the exceptions of soup, beef and chicken, 12 out of 15 types of drinks or foods were found to add to the total calories ingested in evening meals without displacing calories ingested in other forms, while ingestion of non-caloric diet sodas was not associated with differences in intake. The fat and protein, but not carbohydrate, contents of the items correlated with a measure of the satiating properties of the particular food or beverage, namely the correlation between the amount ingested of the particular type and the amounts of other nutrients ingested in the meal. The results confirm that intake at a meal is quite elastic and can be significantly influenced by the presence or absence of particular components of the meal and their constituents.     

Two-year clinical evaluation of preventive resin restorations

Structural and morphologic changes in peripheral blood neutrophilic leukocytes were examined with the aim to diagnose and predict the clinical course of acute pyoinflammatory maxillofacial diseases. Classification of structural and morphologic signs of neutrophilic leukocyte activation levels during odontogenic pyoinflammatory diseases were studied using optic and electron microscopy. There degrees of neutrophilic leukocyte activation were distinguished: inactive, moderately active, and hyperactive. Inactive cells occurred in normal subjects and patients with mild forms of the disease, moderately active ones in patients with mild and medium-severe condition, hyperactive ones in those with medium-severe and grave disease. Increased count of hyperactive neutrophilic leukocytes in the peripheral blood was a sign predicting a grave clinical course of the disease with complications.     

In vivo efficacy of antimicrobial-coated fabric from prosthetic heart valve sewing rings

Escherichia coli tRNA(Val) with pyrimidine substitutions for the universally conserved 3'-terminal adenine can be readily aminoacylated. It cannot, however, transfer valine into polypeptides. Conversely, despite being a poor substrate for valyl-tRNA synthetase, tRNA(Val) with a 3'-terminal guanine is active in in vitro polypeptide synthesis. To better understand the function of the 3'-CCA sequence of tRNA in protein synthesis, the effects of systematically varying all three bases on formation of the Val-tRNA(Val):EF-Tu:GTP ternary complex were investigated. Substitutions at C74 and C75 have no significant effect, but replacing A76 with pyrimidines decreases the affinity of valyl-tRNA(Val) for EF-Tu:GTP, thus explaining the inability of these tRNA(Val) variants to function in polypeptide synthesis. Valyl-tRNA(Val) terminating in 3'-guanine is readily recognized by EF-TU:GTP. Dissociation constants of the EF-Tu:GTP ternary complexes with valine tRNAs having nucleotide substitutions at the 3' end increase in the order adenine < guanine < uracil; EF-Tu has very little affinity for tRNA terminating in 3' cytosine. Similar observations were made in studies of the interaction of 3' end mutants of E. coli tRNA(Ala) and tRNA(Phe) with EF-Tu:GTP. These results indicate that EF-Tu:GTP preferentially recognizes purines and discriminates against pyrimidines, especially cytosine, at the 3' end of aminoacyl-tRNAs.     

Stress and strain in staphylococcal nuclease

Protein molecules generally adopt a tertiary structure in which all backbone and side chain conformations are arranged in local energy minima; however, in several well-refined protein structures examples of locally strained geometries, such as cis peptide bonds, have been observed. Staphylococcal nuclease A contains a single cis peptide bond between residues Lys 116 and Pro 117 within a type VIa beta-turn. Alternative native folded forms of nuclease A have been detected by NMR spectroscopy and attributed to a mixture of cis and trans isomers at the Lys 116-Pro 117 peptide bond. Analyses of nuclease variants K116G and K116A by NMR spectroscopy and X-ray crystallography are reported herein. The structure of K116A is indistinguishable from that of nuclease A, including a cis 116-117 peptide bond (92% populated in solution). The overall fold of K116G is also indistinguishable from nuclease A except in the region of the substitution (residues 112-117), which contains a predominantly trans Gly 116-Pro 117 peptide bond (80% populated in solution). Both Lys and Ala would be prohibited from adopting the backbone conformation of Gly 116 due to steric clashes between the beta-carbon and the surrounding residues. One explanation for these results is that the position of the ends of the residue 112-117 loop only allow trans conformations where the local backbone interactions associated with the phi and psi torsion angles are strained. When the 116-117 peptide bond is cis, less strained backbone conformations are available. Thus the relaxation of the backbone strain intrinsic to the trans conformation compensates for the energetically unfavorable cis X-Pro peptide bond. With the removal of the side chain from residue 116 (K116G), the backbone strain of the trans conformation is reduced to the point that the conformation associated with the cis peptide bond is no longer favorable.     

Cross-reactivity of workshop antibodies with cells from domestic and wild ruminants

Reactivities of the monoclonal antibodies (mAbs) from the workshop panel with cells from cattle, sheep, goats, Cape buffalo (Syncerus caffer) and waterbuck (Kobus defassa) were tested. One hundred and sixty-nine mAbs reacted with bovine cells and 111 with sheep cells; 86 were shown to react with goat cells, 71 with buffalo cells and 70 with waterbuck cells. Some mAbs cross-reacted with all five ruminants tested, and are likely to react with epitopes that are conserved in other ruminant species. Such mAbs will obviate the need to produce mAbs panels to leukocyte antigens of other ruminants.     

Chemical and pharmacological studies of Phyllanthus caroliniensis in mice

We studied retrospectively the effect of long-term treatment with an inhaled corticosteroid on bronchial hyperresponsiveness (BHR) and clinical asthma in moderate-severe asthmatic subjects. Fifty-eight patients who had used beclomethasone dipropionate (BDP) over one year, were enrolled in this study. BHR was measured before and after treatment with BDP by the methods recommended by Japanese Society of Allergology. Moreover we examined the clinical factors and the frequency of acute exacerbations. The results as follows: 1) The mean age was 48.8 years and the mean asthma history was 9.2 years. The mean dose and mean time of BDP administration was 801 micrograms/day and 28.1 months, respectively. 2) Patients during BDP treatment over one year showed about 6-fold mean improvements in BHR, but there were many patients who showed no improvements in BHR. 3) We retrospectively divided all the patients into two groups. Namely, the improved group (n = 25) showed more than 4 fold improvement in BHR and unchanged group (n = 33), less than 4-fold. But there were no significant differences in clinical characteristics and %FEV1 during treatment with BDP. 4) The unchanged group had more near fatal episodes in the past than the improved group. 5) There was significant decrease in acute exacerbation during treatment with BDP, but the unchanged group had more acute exacerbations than the improved group during treatment with BDP. These results indicates that there are many patients who had no improvement on BHR with long term BDP treatment and they have more acute exacerbations due to various stimuli. In conclusion, asthma is recognized chronic inflammatory disease and inhaled corticosteroid therapy has been recommended as the first line therapy. We must further study the clinical problems and underlying mechanisms concerning about treatment with an inhaled corticosteroid.