Detection of Pneumocystis carinii in lung-derived samples using monoclonal antibodies to an 82 kDa parasite component

Monoclonal antibodies against Pneumocystis carinii (PC) antigenic epitopes resistant to denaturing procedures were developed by immunization of mice with isolated parasites and a urea extract of infected human lung tissue. Paraffin sections of infected lung tissue were used as antigen in the screening for reactive clones. The target antigen was identified as an 82 kDa parasite specific component in immunoblotting. The antibody showed no cross-reactivity with human lung tissue and various rat tissues and failed to react with a number of parasites and fungi. The antigenic epitope recognized by the anti-82 kDa component was resistant to denaturing procedures involved in fixation and processing of tissues for histology. The antibodies could be used for identification of both cysts and trophozoites in fixed smears of infected lung tissue and bronchoalveolar lavage fluid from infected individuals.     

Development, characterization and use of monoclonal antibodies against sTRAIL: measurement of sTRAIL by ELISA.

Two monoclonal antibodies against tumor necrosis factor (TNF)-related apoptosis inducing ligand (TRAIL), designated VI10E and III6F, have been generated. These antibodies were useful in flow cytometry analysis, immunohistochemistry, immunoprecipitation and in the development of an immunoassay for the detection of soluble TRAIL (sTRAIL)in biological samples. The immunoassay was based on two monoclonal antibodies against TRAIL. VI10E was used as the capture antibody and bound TRAIL was detected with anti-TRAIL from R&D Systems which was digoxigenin (DIG)-labeled. This enzyme-linked immunosorbent assay (ELISA) was specific for TRAIL since a panel of other cytokines did not affect the signal. The immunoassay was suitable for the detection of sTRAIL in human serum and plasma samples, cell culture supernatants and cell lysates. In a preliminary screening, it was found that serum samples from human immunodeficiency virus (HIV)-infected patients contained sTRAIL, and all these positive samples were found in the AIDS group. Using the immunoassay, it was found that phytohaemagglutinin (PHA)-stimulated peripheral blood mononuclear cells (PBMC) to produce significant amounts of sTRAIL, the levels of which increased with exposure time. Thus, the immunoassay for TRAIL presented here represents a useful tool for measuring sTRAIL in various biological samples. It will also permit studies of release mechanisms as well as possible functions of the soluble form of this molecule.     

An alternative method for calculating changes in arterial calf blood flow at rest

Summary In 20 patients we studied the changes in calf arterial blood flow (AF) following spinal and epidural blocks, using venous occlusion strain gauge plethysmography (SGP). AF was calculated both in the conventional way by drawing a tangent to the initial upslope of the curve, and by a new method which measures the time to the point when 50% of venous capacity is reached (tVC₅₀). The statistical differences within and between the spinal and epidural groups for AF and tVC₅₀ were determined. In measuring the post-block changes in AF as compared with the control values, a statistically significant correlation (r_s=–0.85, p<0.01) was observed between the two methods. We conclude that the new variable, tVC₅₀, seems to be potentially useful in calculating changes in arterial blood flow at rest, and could be used, for example, in connection with surgical and pharmacological interventions, especially when the initial upslope of the SGP curve is equivocal.     

Benefits of bilateral electrical stimulation with the nucleus cochlear implant in adults: 6-month postoperative results.

OBJECTIVE: To evaluate the benefits of bilateral electrical stimulation for hearing-impaired adult subjects using the Nucleus 24 cochlear implant in a multicenter study, and to compare and quantify performance on speech perception measures in quiet and in noise and localization ability for unilateral and bilateral cochlear implant use. DESIGN:: Repeated single subject measures were carried out for each subject, with each subject serving as their own control. Assessment of unilateral and bilateral listening conditions for performance on tests of speech comprehension and sound localization were performed. Speech comprehension measures were performed in quiet at 0 degree azimuth and in the presence of background noise simultaneously presented from the same speaker and spatially separated by 90 degrees, at S+45 degrees N45 degrees and at S-45 degrees N+45 degrees . Test materials included Freiburger monosyllabic words, Oldenburger sentences, and the Hochmair-Schulz-Moser sentences. Tests of localization were performed in the horizontal plane with 12 speaker locations 30 degrees apart using a shortened sentence stimulus from the Hochmair-Schulz-Moser sentences at two possible presentation levels of 55 and 70 dB sound pressure level for assessment of directionality. The binaural advantage provided by bilateral stimulation was calculated with respect to each ear separately, classified as either the better or poorer performing ear for each speech material in quiet and in noise test conditions. For localization of sound, the binaural advantage was compared with left and right ears separately. Paired comparisons for performance data in all conditions were carried out by considering measurements for each subject in different conditions as paired observations and applying the Student's t test to determine the statistical difference between the data sets. SETTING: Tertiary referral centers with a cochlear implant program. PATIENTS: Thirty-seven profoundly hearing-impaired adults were enrolled in the study, 22 simultaneously and 15 sequentially bilaterally implanted. All patients received the Nucleus 24 cochlear implant and used the Nucleus SPrint or ESPrit 3G speech processor, with the vast majority using the ACE speech coding strategy. RESULTS: For spatially separated speech in noise conditions, an interaural performance advantage for the ear closest to the speech source (i.e., with a superior signal to noise ratio) compared with that for the ear closest to the noise source (i.e., with an inferior signal to noise ratio) is consistently demonstrated regardless of whether it is the better or poorer performing ear closest to the speech signal. This is referred to as a significant binaural head-shadow benefit, resulting in a mean improvement between -10 dB and -11.4 dB in the critical signal to noise ratio required for 50% speech comprehension for the Olden-burger sentences and a mean improvement in the maximum score of 42% to 55% for the ear closest to the speech signal over the ear farthest away for the Hochmair-Schulz-Moser sentences. Bilateral stimulation is always observed to provide a performance advantage over the unilateral listening condition for either ear when ipsilateral to the noise source. In addition, as demonstrated by approximately half the subjects tested in noise with the Hochmair-Schulz-Moser sentences, a performance advantage of bilateral stimulation may be observed over the better ear alone when positioned ipsilateral to the speech signal, which is referred to as a binaural squelch effect. On average, for the group, this resulted in a statistically significant improvement in speech comprehension scores of 8% in the bilateral listening condition compared with the scores for the better ear alone. Through assessment of comprehension of coincidental speech in noise and speech in quiet, a significant benefit of binaural redundancy was noted for the group for Oldenburger sentence scores in noise and in quiet compared with unilateral scores for either ear and for the Freiburger monosyllabic words in quiet in comparison with the better ear alone scores. Binaural stimulation also led to a significant improvement in localization ability over either monaural condition, with the root mean square degrees of error reduced by 38 degrees compared with that observed for unilateral stimulation. CONCLUSION: Similar to what has been observed for bilateral acoustic stimulation in the past, bilateral electrical stimulation provides the foundation for the potential advantages of the head-shadow effect, providing a binaural head-shadow benefit and binaural auditory processing such as binaural redundancy and binaural squelch effects, all of which combine to lead to improved speech comprehension over unilateral listening conditions. The combination of improved speech comprehension and improved localization ability made available through bilateral electrical stimulation provides the necessary foundation to further assist the hearing-impaired listener to better cope with communication in the everyday listening situation both in noise and in quiet.     

A new L-shaped titanium prosthesis for total reconstruction of the ossicular chain.

OBJECTIVE: To describe the technique used for total ossiculoplasty with the Fisch titanium total prosthesis and evaluate the 1-year postoperative functional results in patients presenting with the stapes (or footplate) without the malleus handle. STUDY DESIGN: Prospective trial with preoperative and postoperative comparison. SETTING: Academic and private practice tertiary care center for otologic surgery. PATIENTS: Forty-nine consecutive patients operated on from September 1996 to December 2000. SURGERY:: Staged ossicular reconstruction with a Fisch titanium total prosthesis placed between the footplate and the tympanic membrane without regard to the presence or absence of the stapes arch. Coupling of the prosthesis to the footplate was achieved by various techniques, including perforation, foot and spike on the footplate without perforation, and shaft alone (without foot) with tragal cartilage fixation (disc or small wedges). There was no interposition of cartilage between the prosthesis head and the tympanic membrane. MAIN OUTCOME MEASURES: Pre- and postoperative air and bone-conduction thresholds and air-bone gaps for pure-tone averages of three and four frequencies and for single frequencies. RESULTS: Postoperative air-bone gap closures within 20 dB distributed equally (50%) between 0.5, 1, and 4 kHz and reached the highest rate (89%) at 2 kHz (p < 0.05). The postoperative air-bone gaps for pure-tone averages reached 0 to 20 dB in 57% and 0 to 30 dB in 87% of the cases. There were no dead ears and no partial or total extrusions of prostheses. The best functional results were achieved through perforation coupling of the spiked foot to the footplate in large oval windows and after fixation of the shaft (without foot) with tragal cartilage disc in narrow oval windows. CONCLUSION: The functional results of the L-shaped Fisch titanium total prosthesis implanted in ears with the stapes but no malleus handle are best at 2 kHz and better than those of comparable columellar titanium prostheses over the remaining tested frequencies.     

Glutathione and phospholipid depletion of liver tumors after arterial ischemia

Breakdown of membrane phospholipids is a causative event leading to irreversible cell injury after ischemia and reperfusion insults, which might be one mechanism leading to liver tumor cell death after repeated arterial ischemia as well. After 2 hr of hepatic dearterialization followed by 30 min of reperfusion tumor phospholipid was measured chromatographically, glutathione (GSH) analyzed by determining nonprotein sulfhydryl and activity of glutathione-S-transferase (GST) determined spectrophotometrically using 1-chloro-2,4-dinitrobenzene (CDNB) as the substrate. A transient, arterial ischemia for 2 hr induced a substantial decrease of phosphatidylserine (PS) and phosphatidylinosital (PI) compared with sham treatment (P < 0.01). Although phosphatidylcholine (PC) and phosphatidylethanolamine (PE) did not significantly decline after a single arterial ischemia for 2 hr, they dropped dramatically following repeated arterial ischemia for 2 hr during 5 days (P < 0.01 and P < 0.05 respectively). GSH was depleted in tumors after both a single (P < 0.01) and repeated arterial ischemia (P < 0.05) and GST was inactivated as well (P < 0.001). By contrast, neither liver phospholipid nor liver GSH or GST was significantly changed. Tumor growth was significantly retarded in rats subjected to repeated arterial ischemia compared with sham treatment (P < 0.01). Repeated arterial ischemia facilitated degradation of tumor membrane phospholipids and induced depletion of GSH and inactivation of GST without affecting the normal liver. Thus, ischemia/reperfusion induced depletion of membrane phospholipids and of GSH might represent two mechanisms by which repeated arterial ischemia led to tumor growth delay. © 1996 Wiley-Liss, Inc.