A novel method for DEAE-dextran mediated transfection of adherent primary cultured human macrophages

The effects of regional and global ischemia on cellular electrical activity and on arrhythmias induced by reperfusion were studied at different Mg2+ concentrations (Mg2+o, 0, 1.2, and 4.8 mM) in perfused rat hearts. Surface electrograms and transmembrane potentials were recorded during control, 10 min of ischemia (perfusion arrest or coronary ligation), and reperfusion. Increasing Mg2+o from 0-4.8 mM decreased heart rate, did not alter action potential morphology, and had a strong antiarrhythmic action on reperfusion following coronary ligation. At low and normal Mg2+o, the incidence of tachyarrhythmias was between 70 and 80%. Global ischemia led to progressive atrioventricular block and the final ventricular beating rate was similar at all Mg2+o despite unequal initial values. The severity of arrhythmias was similar to that found after regional ischemia in Mg2+o = 0, but much lower at normal and high Mg2+o. The resting depolarization induced by coronary ligation decreased as Mg2+o was raised, but such a relation was not seen during global ischemia where the depolarization was less marked. The action potential duration did not vary with the ventricular rate between 160 and 380 beats per min but increased considerably when sinus rate was markedly slowed (40 to 80 bpm) by raising Mg2+o to 9.6 mM. Our data show that a high Mg2+o exerts a strong protection against reperfusion arrhythmias regardless of the type of ischemia. Modulation of the sinus rhythm by Mg2+ may contribute to its protective effect by decreasing K+o accumulation and Na+i loading during ischemia.     

Diagnostic and prognostic relevance of malignancy-associated changes in cervical smears

OBJECTIVE: For approximately 15 years, malignancy-associated changes (MACs) have been consistently found by means of high-resolution cytometry in different tissues, especially in visually normal appearing cervical cells. Their biologic nature is not yet fully understood. The aim of this investigation was to assess the expression of MACs in cervical smears and to evaluate their prognostic relevance. STUDY DESIGN: This study was performed on normal intermediate cells obtained from 53 cytologically positive and 78 cytologically negative cervical smears. From a second sample, 31 cases showing negative cytology were selected for a prospective longitudinal study. Densitometric and texture features were generated, and MACs were described on the basis of multivariate discriminant analysis. RESULTS: Discrimination between positive and negative cases was possible, with a correct classification rate of approximately 80%. After a mean period of 29.5 months, we noted no statistically significant increase in the incidence of cervical intraepithelial neoplasia in the group of healthy but MAC-positive women as compared to those who were MAC negative. CONCLUSION: MACs were constantly expressed in the epithelium of the cervix. Although their prognostic relevance remains unclear, MACs play an important role in the effort to automate cervical cytology.     

The sensory innervation of the hip joint--an anatomical study

Typically obturator nerve blockade is used to relieve hip pain. It sometimes only has a minor effect in resolving symptoms. This clinical observation led us to examine comprehensively the sensory nerve innervation of formalin-fixed hip joint capsules. Following macroscopic preparation, the area of the hip joint capsule was inspected with the aid of an operating microscope. We discovered a separation between the anterior and posterior sensory innervation of the hip joint capsule. The anteromedial innervation was determined by the articular branches of the obturator n. Additionally, the anterior hip joint capsule was innervated by sensory articular branches from the femoral n. In the posterior part we found articular branches from the sciatic n., which in addition to the articular branches from the nerves to the quadratus femoris m., innervate the postero-medial section of the hip joint capsule. Moreover, articular branches of the superior gluteal n. were found, which innervate the posterolateral section of the hip joint capsule. This anatomical study demonstrates that the obturator n. block is insufficient for the treatment of hip pain. Further investigations will determine if these nn. can be reached percutaneously. Effective neural blockade of the hip joint must include the femoral n., the sciatic n. and the superior gluteal n.     

Revised cutoff values of serum aminotransferase in detecting viral hepatitis among CAPD patients: experience from Taiwan, an endemic area for hepatitis B

BACKGROUND: To determine the best cutoff values of aspartate aminotransferase (AST) and alanine amino-transferase (ALT) in detecting viral hepatitis C infection among patients of continuous ambulatory peritoneal dialysis (CAPD). METHODS: 90 (44 male and 46 female) CAPD patients and 526 adult controls (266 male, 260 female) were enrolled. Serum AST and ALT were measured by an auto-analyser monthly. Serum HBsAg was examined using a RIA method and anti-HCV by an second-generation EIA method. The best cutoff values of AST and ALT for detecting viral hepatitis were obtained from the ROC (receiver-operating characteristic) curve. RESULTS: The prevalence of anti-HCV(+) was significantly higher in CAPD patients (16.7%) than in normal controls (4.9%), while that of HBsAg(+) was similar in both groups. CAPD patients had significantly lower levels of serum aminotransferases compared to normal controls. Mean AST were 23.8 IU/l in normal control and 18.8 IU/l in the CAPD patients (P < 0.001). Mean ALT were 21.9 IU/l in normal controls and 15.3 IU/l in the CAPD patients (P < 0.001). CAPD patients with HCV infection had higher serum AST and ALT levels than those without. However, HBV infection did not cause significant serum aminotransferase elevation in patients. The conventional cutoff values of AST (40 IU/l) and ALT (40 IU/l) for detecting viral hepatitis yielded only a sensitivity of 27.3 and 18.2% respectively; on the contrary, our revised cutoff values of AST (24 IU/l) and ALT (17 IU/l) had better sensitivities (AST, 72.7%; ALT, 63.6%). For serial aminotransferase values, the sensitivity of AST and ALT for detecting HCV were 36.4 and 27.3% by conventional criteria, and were both 81.8%, by our newly revised criteria. CONCLUSIONS: Serum aminotransferase cutoff values should be modified for screening viral hepatitis in a CAPD population. Our new cutoff criteria had important clinical implications in providing benefits of earlier detection and possible prevention from chronic hepatic deteriorations.     

Toward targets for initiation of chronic dialysis

OBJECTIVES: To better define the targets for initiation of chronic dialysis, we compared the relationship between the normalized protein equivalent of nitrogen appearance (nPNA, g/kg standard weight/day) and weekly urea clearance (Kt) normalized to total body water (V) in predialysis chronic renal failure (CRF) patients and in patients on continuous ambulatory peritoneal dialysis (CAPD) and hemodialysis (HD). We also studied the relationships of other nutritional parameters to weekly Kt/Vurea in CRF patients. DESIGN: This cross-sectional study was a prospective observational design meant to study each patient once. SETTING: The University Hospital and Clinics and Harry S. Truman VA Medical Center, Columbia, Missouri. PATIENTS: Forty-five consecutive predialysis CRF patients were enrolled and the results compared with patients on CAPD and HD. RESULTS: In CRF, the nPNA calculated from urea appearance correlated with the weekly Kt/Vurea (r = 0.57, p < 0.0001) and, using exponential best-fit, nPNA = 1.217 x (1-e-0.769Kt/V). This exponential relationship was similar to that for CAPD and both were different from that in patients on HD. Likewise, nPNAs, calculated from Kjeldahl nitrogen output, and weekly Kt/Vurea were correlated (r = 0.37, p = 0.014) and, using exponential best-fit, nPNA = 1.102(1-e-0.867Kt/V), similar to the relationship in patients on CAPD. Evidence is presented that these relationships are not explained only by mathematical coupling. There was a significant correlation between the weekly Kt/Vurea and 24-hour urinary creatinine excretion. CONCLUSIONS: The findings suggest that in CRF, as in CAPD, a weekly Kt/Vurea less than 2.0 is likely to be associated with a nPNA less than 0.9 g/kg standard weight. In CRF patients, initiation of chronic dialysis should be considered if weekly renal Kt/Vurea falls below 2.0 and a nPNA greater than 0.8 is desired.     

Monitoring of impurities using high-performance liquid chromatography with diode-array detection: eigenvalue plots of partially overlapping tailing peaks

The chromatogram of ropinirole in the presence of about 5% of a closely eluting impurity, obtained by HPLC with diode-array detection, was analysed by chemometric procedures. Log eigenvalue plots were used to determine the relative composition of regions of the chromatogram. It is shown that since the peaks exhibit tailing, unusual behaviour is found in the plots. This is verified by performing simulations, in which it is demonstrated that peak asymmetry has a pronounced influence on this chemometric approach. In many cases of liquid chromatographic analysis, asymmetric peak shapes are encountered and methods for peak purity assessment should be re-evaluated in the light of these asymmetries.     

Quenching of chlorophyll a fluorescence in the aggregates of LHCII: steady state fluorescence and picosecond relaxation kinetics

The protein composition, steady state and time-resolved fluorescence emission spectra were studied in solubilized and aggregated LHCII complexes, that were prepared according to two different isolation protocols: (1) by fractionation of cation-depleted thylakoid membranes using the non-ionic detergent Triton X-100 according to the procedure of Burke et al. [(1978) Arch. Biochem. Biophys. 187, 252-263] or (2) by solubilization with N-beta-dodecyl maltoside (beta-DM) of photosystem II (PSII) membrane fragments in the presence of cations [Irrgang et al. (1988) Eur. J. Biochem. 178, 207-217]. Based on the analysis of the decay-associated emission spectra measured at 10 and 80 K five long-wavelength chlorophyll species were identified in aggregated LHCII complexes. These five forms are characterized by emission maxima at 681.5, 683, 687, 695, or 702 nm. All of these forms were found in both types of LHCII preparations but the relative amounts and temperature dependency of these species were markedly different in the aggregated LHCII complexes isolated by the two procedures. It was found that these differences cannot be simply explained by effects due to using a less mild detergent as beta-DM or by an ionic influence of Ca2+. Biochemical analysis of the protein composition showed that beta-DM type LHCII consists of all the chlorophyll (Chl)binding proteins belonging to the antenna system of PSII except the CP29 type II gene product (CP29). In contrast, the Triton X-100-solubilized LHCII is highly depleted in CP26 (CP 29 type I gene product) and is contaminated by a variety of unidentified polypeptides. It is proposed that the aggregates of LHCII prepared using Triton X-100 acquire specific spectral and kinetic features due to interaction between the bulk of LHCII subunits and minor protein(s).