Evaluation of PAF antagonists using human neutrophils in a microtiter plate assay

This paper describes a testing model for the detection and evaluation of PAF antagonists, based on the inhibition of PAF-elicited elastase release by human neutrophils. Incubations are performed in microtiter plates in the presence of a specific fluorogenic elastase substrate allowing direct measurement of the exocytosis response by means of a 96-well fluorescence reader. Determinations of the IC50 values for five established PAF antagonists, Ro 19-3704, BN 52021, CV-3988, 48740 RP and kadsurenone, showed that the new model is comparable in sensitivity and discriminative capacity to other in vitro assays. From the effect of antagonists on the PAF concentration-response curve pA2 values could be calculated and information on the type of antagonism obtained. BN 52021 was found to behave as a competitive antagonist while Ro 19-3704 showed a more complex type of inhibition. As a one-plate system, the test is simple to handle and highly reproducible, and appears therefore particularly useful for large drug screening programs.     

MGA5 Health Utilities Index: Further Evidence of Responsiveness

Quality adjusted life years (QALYs) are well recognized as a valid measure for outcomes in cost-effectiveness analyses. However, it is difficult to obtain a summary utility score from health status measure such as the SF-36.
OBJECTIVE: To predict a summary utility score (represented by HUI) from the scores on the SF-36.
METHODS: A structural equation framework was applied to data collected from 1992 to 1995 on the Southern California Kaiser Permanente population (n = 5,794). An instrumental variable (IV) method mitigated the endogeneity in estimating the HUI(MarkII). Socioeconomic and disease variables were used as covariates. A split-sample analysis provided cross-validation.
RESULT: This model predicted 33.68% of the observed variance in HUI index scores with an adjusted R²of 0.3335. Observed HUI index scores were distributed with a mean of 0.7963 and std. deviation of 0.1796. Parameter estimates of most of the SF-36 components (except General Health & Social Functioning) showed statistical significance at α= 0.05 level. People with high chronic disease scores were found to have low SF-36 scores, and parameter estimates of this covariate were also found statistically significant at α= 0.05 level in all structural equations. However, all the socioeconomic variables showed statistical insignificance. Comparison of "Forecasting" and "Estimation" sub-samples showed satisfactory results during cross-validation.
CONCLUSION: Result of this study provides a quantitative link between two important measures of health status. The present model can be used to estimate overall health utility summary scores from previous studies using the SF-36.     

Pain after barium enema: effect of CO2 and air on double-contrast study

One hundred fifty-one consecutive patients scheduled for double-contrast barium enema studies were assigned randomly to insufflation with either air or carbon dioxide (CO2) in a double-blind, prospective trial. Within 24 hours after the enema study, the patients were contacted by telephone by an interviewer, who completed a standard questionnaire. Radiographs from the enema studies were assessed for quality by two radiologists. Pain experienced after the procedure was graded from 0 (none) to 4 (severe). Clinically relevant (grades 2-4) pain was experienced by 30% of patients after insufflation with room air, compared with 11% of patients in whom CO2 was used for insufflation (P = .005). The mean pain score for CO2 was 0.4, and for room air, 1.2 (P less than .005). Although five patients experienced grade 4 pain after insufflation with air, no patient reported severe pain after CO2 insufflation. Post-evacuation films confirmed there was significantly less residual gas in the CO2 group. The quality of radiographs was equal in the two groups. CO2 has advantages for use in the double-contrast barium enema examination.     

Excretory urography with iohexol: evaluation in children

A multicenter clinical study was conducted using iohexol, a second-generation nonionic contrast medium, for excretory urography performed in 130 children. Doses of iohexol (300 mg iodine/ml) ranged between 150 and 660 mgI/kg (0.5 and 2.2 ml/kg). Iohexol was tolerated well, and no significant adverse reactions occurred. Sixty-five iohexol urograms were evaluated to determine the minimum dose for adequate visualization of the kidneys and collecting systems. A dose greater than 300 mgI/kg (1.0 ml/kg) always resulted in a urogram of diagnostic quality, while visualization was insufficient for diagnosis in 10% of studies done with doses of 150-300 mgI/kg (0.5-1.0 ml/kg). Another 65 iohexol urograms were compared in a blinded manner with a similar number of studies performed using iothalamate meglumine at comparable iodine concentration and dose. Visualization of calyces and pelvoinfundibular structures achieved with iohexol was rated better with statistical significance, but there was no difference in visualization of the renal parenchyma or ureters. Use of iohexol in excretory urography may be advantageous in children who are at greatest risk for an adverse reaction to contrast media or in those most likely to benefit from use of a low osmolality contrast agent.     

Interobserver variation in the interpretation of abdominal radiographs

A total of 140 sets of abdominal radiographs were reviewed independently by four qualified diagnostic radiologists. The degree of interobserver agreement was determined by calculating kappa values for 19 commonly used radiographic signs and diagnoses. There was fair to excellent interobserver agreement for 11 signs and diagnoses and poor agreement for the remaining eight. The signs and diagnoses for which agreement is poor cannot be considered reliable and include particularly large bowel obstruction and nonspecific gas pattern.     

B-CLL cells are capable of synthesis and secretion of both pro- and anti-angiogenic molecules.

Initial work has shown that clonal B cells from B-chronic lymphocytic leukemia (B-CLL) are able to synthesize pro-angiogenic molecules. In this study, our goal was to study the spectrum of angiogenic factors and receptors expressed in the CLL B cell. We used ELISA assays to determine the levels of basic fibroblast growth factors (bFGF), vascular endothelial growth factor (VEGF), endostatin, interferon-alpha (IFN-alpha) and thrombospondin-1 (TSP-1) secreted into culture medium by purified CLL B cells. These data demonstrated that CLL B cells spontaneously secrete a variety of pro- and anti-angiogenic factors, including bFGF (23.9 pg/ml +/- 7.9; mean +/- s.e.m.), VEGF (12.5 pg/ml +/- 2.3) and TSP-1 (1.9 ng/ml +/- 0.3). Out of these three factors, CLL B cells consistently secreted bFGF and TSP-1, while VEGF was expressed in approximately two-thirds of CLL patients. Of interest, hypoxic conditions dramatically upregulated VEGF expression at both the mRNA and protein levels. We also employed ribonuclease protection assays to assay CLL B cell expression of a variety of other angiogenesis-related molecules. These analyses revealed that CLL B cells consistently express mRNA for VEGF receptor 1 (VEGFR1), thrombin receptor, endoglin, and angiopoietin. Further analysis of VEGFR expression by RT-PCR revealed that CLL B cells expressed both VEGFR1 mRNA and VEGFR2 mRNA. In summary, these data collectively indicate that CLL B cells express both pro- and anti-angiogenic molecules and several vascular factor receptors. Because of the co-expression of angiogenic molecules and receptors for some of these molecules, these data suggest that the biology of the leukemic cells may also be directly impacted by angiogenic factors as a result of autocrine pathways of stimulation.     

Isolated isochromosome 17q: a distinct type of mixed myeloproliferative disorder/myelodysplastic syndrome with an aggressive clinical course

A clinicopathologic study was performed on 15 patients with haematological malignancies in which isochromosome 17q [i(17q)] was the sole structural chromosome abnormality identified in bone marrow. The data indicated that an isolated i(17q) is associated with a distinct type of mixed chronic myeloproliferative/myelodysplastic disorder with an aggressive clinical course. The patients ranged in age from 37 to 83 years (median 60) with a M:F ratio of 3:1. All cases were chronic myeloid disorders with mixed proliferative and dysplastic features, making classification difficult. 11 patients tested for BCR/ABL gene fusion were normal. A low bone marrow blast count (<5%) at presentation was a typical finding. All cases had severe myeloid dysplasia which included non-segmented neutrophils and an increase in the monocyte/macrophage lineage. Fluorescence in situ hybridization (FISH) analysis of one case showed the i(17q) to involve all myeloid lineages, but not the lymphocytes. For cases with complete follow-up (n = 11) the median survival was 2.5 years (range 0.83-5.25) and 64% progressed to AML prior to death. The following features were identified which defined the haematological disorder associated with an isolated i(17q): (1) adult patient, (2) chronic myeloid disorder with clonal involvement of all myeloid lineages, (3) mixed chronic myeloproliferative/myelodysplastic features, (4) severe hyposegmentation of neutrophil nuclei, (5) prominence of the monocyte/macrophage lineage, (6) high risk for progression to AML, and (7) median survival of 2.5 years.     

Comparative Trial of Norfloxacin and Macrocrystalline Nitrofurantoin (Macrodantin) in the Prophylaxis of Recurrent Urinary Tract Infection in Women

SUMMARY Eighty-eight women with a history of recurrent urinary tractinfection (at least four attacks in the preceding 12 months)were randomized to take either norfloxacin 200 mg at night (45patients) or macrocrystalline nitrofurantoin 100 mg at night(43 patients) for 12 months. A decrease in the number of symptomaticattacks while taking this prophylaxis was observed in 94 percent of the patients and this improvement was maintained duringthe 6 months following the end of prophylaxis in 69 per cent.The mean interval between symptomatic episodes while takingprophylaxis was 7.2-fold and 6.9-fold greater, respectively,than in the 12 months before starting prophylaxis. There wereonly nine breakthrough infections during 74 patient-years ofprophylaxis, four in patients taking norfloxacin (two enterococci,one Staphylococcus epidermidis, one Escherichia coli), and fivein those taking macrocrystalline nitrofurantoin (four E. coli,one Klebsiella pneumoniae). Adverse events caused four patientstaking norfloxacin (8 per cent) and seven taking macrocrystallinenitrofurantoin (14 per cent) to stop prophylaxis. Norfloxacinhad a marked suppressive effect on the coliform part of thefaecal flora, with no emergence of resistance. Thus, norfloxacinappears to be an excellent alternative agent to macrocrystallinenitrofurantoin for the prevention of recurrent urinary infections.     

Stretch reflex adaptation in elbow flexors during repeated passive movements in unilateral brain-injured patients

OBJECTIVE: To evaluate the effects of repeated, externally imposed, flexion-extension movements of the elbow on the resulting stretch reflex response in hemiparetic spastic brain-injured patients. These effects were compared within a recording session and across sessions for the same subject to determine the impact of movement history on the quantification of spastic hypertonia using the stretch reflex response. DESIGN: Twenty to 30 sequential, constant velocity flexion-extension movements were applied to the impaired elbow of our cohort, with a 10-second hold interposed between flexion and extension. Movements were applied regularly at 1-minute intervals. Changes in stretch reflex responses were monitored during the applied movements. PARTICIPANTS: We examined a convenience sample of seven hemiparetic brain-injured subjects between the ages of 26 and 60 yrs, with moderate-to-severe spastic hypertonia of elbow muscles (Ashworth score 2-4/4). Subjects participated in 2 to 9 sessions. MEASURES: Elbow torque, position, velocity, and electromyograms of the biceps, brachioradialis, and triceps muscles were recorded for each flexion and extension movement. Stretch reflex torque was calculated by subtracting passive torque from total elbow torque, recorded over large amplitude movements. A linear regression analysis quantified both the initial torque response of the stretch reflex and the ensuing adaptation of the stretch reflex during sequential movements. Intersession variability was characterized both for spastic hypertonia measures and for stretch reflex adaptation. RESULTS: Repeated, externally imposed, sequential flexion-extension movements of the elbow decreased the elbow flexor stretch reflex in six of seven subjects. The mean reduction in reflex torque after 30 movements was 50% of the initial torque values (p = .001, t test vs. 0% change). Intersession stretch reflex responses for each subject were found to vary greatly (SDs of reflex torque ranged from 0.1 to 4.0 Nm), and there were also significant variations in the degree of adaptation between subjects. CONCLUSIONS: Stretch reflex adaptation must be taken into consideration when spastic hypertonia is quantified using repeated joint motion, as is often the case. The magnitude of intersession variation in spastic hypertonia measures suggests that ideally, such measurements should be made across multiple sessions before conclusions are made regarding the efficacy of spastic hypertonia interventions. This study provides quantitative evidence that repeated joint movements may have a significant short-term beneficial effect on spastic hypertonia.