Cerebral phaeohyphomycosis caused by a dematiaceous scopulariopsis species.

This report describes a rapidly fatal case of cerebral phaeohyphomycosis in a 33-year-old immunocompetent male. The infection presented as a single large lesion in the deep white matter of one temporal lobe, which was then removed surgically. Histologic features observed in the lobectomy specimen were characterized by perivascular sleeves of mononuclear cells accompanied by hemorrhages. These were reminiscent of acute hemorrhagic leukoencephalitis except for the presence of rare fungal organisms and sparse multinucleated giant cells similar to those occurring in AIDS. During the four days following surgery, a large focus of cerebritis with massive invasion of fungi developed in each centrum semiovale around the ventriculostomy sites. Fungal culture of the brain obtained at autopsy grew an organism consistent with a Scopulariopsis species.

Comparison of capillary electrophoresis and polyacrylamide gel electrophoresis for the evaluation of T and B cell clonality by polymerase chain reaction.

Polymerase chain reaction (PCR) technique is widely used in the diagnosis of lymphoma, and PCR amplification products are typically detected by polyacrylamide gel electrophoresis (PAGE). However, the identification of small clonal populations, or the distinction of clonal PCR products in a polyclonal milieu remains difficult, requiring technically demanding alterations to gel analysis. This study describes an alternative approach using a capillary electrophoresis (CE) system to produce an accurately sized electropherogram. A variety of patient samples were examined, including solid tissue, peripheral blood, bone marrow aspirates, and paraffin-embedded tissue. A total of 28 samples were evaluated by PCR for B-cell clonality by detection of immunoglobulin heavy chain gene rearrangement and 29 samples for T-cell clonality by detection of T-cell gamma locus gene rearrangement. Standard 10% PAGE analysis of PCR products was compared with CE. There was a 100% concordance in the assessment of both B-cell and T-cell clonality. Dilution studies with the SUP-B15 cell line showed a detection limit of 0.03% for B-cell clonality and 0.05% for T-cell clonality using CE, versus 0.2% to 1%, respectively for PAGE. Automated, fluorescent analysis of PCR products by CE seems to be at least equally as effective as gel-based analysis for the detection of clonal B-cell and T-cell populations. Moreover. CE offers superior resolution and improved sensitivity, thus representing a significant improvement over traditional gel electrophoretic techniques in these regards.

Low molecular weight heparin prevents the pulmonary hemodynamic and pathomorphologic effects of endotoxin in a porcine acute lung injury model.

Tumor necrosis factor alpha (TNF-alpha) activity, platelet and neutrophil degranulation and margination, and increased vascular permeability are central to the pathophysiology of endotoxin-mediated acute lung injury. Nonanticoagulant activities of low molecular weight heparin (LMWH) include solubilization of the TNF-alpha receptor protein, inhibition of neutrophil adhesion, and regulation of thromboxane B2 (TXB2) biosynthesis. In this study, we evaluated the ability of LMWH to modulate TNF-alpha and TXB2 activity during endotoxemia and the subsequent effects on pulmonary hemodynamics. Domestic pigs 8-10 weeks old were anesthetized and catheterized for standard cardiopulmonary measurements and the lungs harvested for cuff:vessel ratio, myeloperoxidase activity, and permeability index. Pigs were randomly assigned to one of four groups: lipopolysaccharide (LPS) (n = 6), given .5 microg/kg/h Escherichia coli LPS intravenously for 6 h; saline control (n = 5); LMWH (n = 5), given .5 mg/kg LMWH for 30 min, followed by .5 mg/kg/h; and LMWH + LPS (same dosages, n = 6). Administration of LPS resulted in increased plasma TNF-alpha and TXB2 activity; increased pulmonary arterial pressure, pulmonary vascular resistance, and alveolar-arterial oxygen tension; decreased systemic arterial oxygen tension; and pulmonary edema. The cardiopulmonary parameters for the LMWH-treated pigs did not differ from those of the saline-treated control pigs. Pretreatment with LMWH attenuated the LPS-mediated TNF-alpha and TXB2 activity and attenuated LPS-mediated pulmonary hypertension, hypoxemia and neutrophil emigration, and edema formation. In conclusion, the data show that the protective effects of LMWH in this model of acute lung injury are associated with altered neutrophil adhesion and TNF-alpha and thromboxane activity.

A comparison of the BioStar Strep A OIA rapid antigen assay, group A Selective Strep Agar (ssA), and Todd-Hewitt broth cultures for the detection of group A Streptococcus in an outpatient family practice setting.

In some studies the BioStar Strep A OIA (optical immunoassay) has yielded inconsistent results, although originally it was reported to be more sensitive than conventional culture for the detection of group A Streptococcus (GAS). The Group A Selective Strep Agar with 5% sheep blood (ssA) incubated anaerobically has been reported to be more sensitive than conventional culture in the detection of GAS. We compared the BioStar Strep A OIA GAS rapid antigen detection kit to anaerobic culture on ssA with and without preincubation in Todd-Hewitt broth (THB) for the detection of GAS. From September 1995 through January 1996, throat swabs were collected in duplicate from 75 children (< or = 18 years) and 188 adults (> 18 years) who presented with pharyngitis in the outpatient University of New Mexico Family Practice Clinic. Thirty-one (12%) of the 263 cases were positive for GAS by culture and/or broth. Compared with anaerobic culture on the ssA, with and without preincubation in THB, the sensitivity, specificity, positive predictive value, and negative predictive value of the BioStar Strep A OIA were 77, 62, 22, and 95%, respectively. Compared with enrichment in THB followed by subculture on ssA and anaerobic incubation, the sensitivity, specificity, positive predictive value, and negative predictive value of direct culture on ssA and anaerobic incubation were 79, 99, 98, and 96%, respectively. All isolates were serologically grouped. The BioStar Strep A OIA is as sensitive as direct culture on ssA incubated anaerobically, but the low specificity and low positive predictive value when the OIA is used in low prevalence populations could lead to unnecessary antibiotic treatment.

Rapid detection of oleander poisoning using fluorescence polarization immunoassay for digitoxin. Effect of treatment with digoxin-specific Fab antibody fragment (ovine).

Poisoning from the oleander plant is common. Taking advantage of the high cross-reactivity of oleandrin, the major cardiac glycoside found in the oleander plant, we demonstrated that the serum digitoxin assay can be successfully used for the rapid diagnosis of oleander poisoning. Digitoxin is rarely used for treatment of cardiac disorders in the United States and has a therapeutic range of 19.7 to 39.3 nmol/L. In a typical oleander poisoning, serum oleandrin concentrations may reach 174 mmol/L or more. A serum specimen supplemented with 174 mmol/L of oleandrin containing no digitoxin showed an apparent digitoxin concentration of 1,272.1 nmol/L, a very high value compared with the range of the serum digitoxin assay, which is 2.6 to 104.8 nmol/L. Moreover, the response of the serum digitoxin assay with serum specimens containing various concentrations of oleandrin (and no digitoxin) is linear. Therefore, the oleandrin concentration in serum can be calculated from the apparent digitoxin concentration to access the severity of poisoning. Recently, the usefulness of the digoxin-specific Fab antibody fragment in the treatment of oleander poisoning has been described; however, no laboratory test was performed to demonstrate the progress of therapy. We demonstrated that the digoxin-specific Fab antibody can bind oleandrin in vitro, thus reducing the pharmacologically active free oleandrin. Because Fab and oleandrin bound to Fab are absent in the protein-free ultrafiltrates, monitoring the activity of free oleandrin in the ultrafiltrates can be used for monitoring the effectiveness of therapy.

A rapid cost-effective high-performance liquid chromatographic (HPLC) assay of serum lamotrigine after liquid-liquid extraction and using HPLC conditions routinely used for analysis of barbiturates.

Lamotrigine (lamictal) is a new anticonvulsant drug approved by the FDA for clinical use. Therapeutic monitoring of lamotrigine is useful for patient management and avoidance of toxicity. The suggested therapeutic range is 1 to 4 micrograms/ml. The authors describe a simple high-performance liquid chromatographic (HPLC) method for analysis of lamotrigine from serum. Serum (0.5 ml) was alkalinized with borate buffer (pH 9.8). Lamotrigine and the internal standard thiopental were extracted with 10 ml of chloroform. After evaporation of the extract, the residue was reconstituted in the mobile phase (prepared by mixing 750 ml of potassium dihydrogen phosphate, 550 ml of deionized water, 430 ml of methanol, and 100 microliters of triethylamine as an ion pairing reagent) and injected into an LC-18 column (15 cm x 4.6 mm). The authors use this HPLC system routinely in their laboratory for the analysis of barbiturates. They demonstrated that the same system can be used for the analysis of lamotrigine. The within-run and between-run precisions of the lamotrigine assay were 1.63% (mean = 3.05, SD = 0.05 microgram/ml, n = 6) and 3.7% (mean = 2.97 micrograms/ml, SD = 0.11, n = 8). The assay was linear for serum lamotrigine concentrations of 0.5 microgram/ml to 20 micrograms/ml with a detection limit of 0.5 microgram/ml. The authors observed excellent correlation between serum lamotrigine concentrations measured by their assay and a reference laboratory in six patients receiving lamotrigine. Their assay is free from interferences from common tricyclic antidepressants, benzodiazepines, other common anticonvulsants, salicylate, and acetaminophen.

Morphometric analysis of oleic acid-induced permeability pulmonary edema: correlation with gravimetric lung water.

The technique used most commonly to quantitate pulmonary edema in in vivo animal models is postmortem gravimetric analysis (wet:dry) ratio. To determine whether lung water can be quantitated morphometrically, as accurately as by the commonly used gravimetric analysis, perivascular edema (cuff) area to vessel area ratio was correlated to wet:dry ratio. Anesthetized pigs were given either oleic acid (20 mg/kg/h, intravenously) or physiologic saline. At 4 h, lungs were excised and cuff:vessel and wet:dry ratio analysis was performed. The intermediate lobe was clamped across its main stem bronchus to maintain peak inspiratory inflation, excised, frozen in liquid nitrogen, and stored at -70 degrees C until cryostat sectioning and quantification of perivascular interstitial edema (cuff) area. Gravimetric analysis (wet:dry ratio) was performed on the remaining lung. Mean cuff:vessel and wet:dry analyzes showed that lung water increased significantly (p < .01) in the oleic-acid treated group (4.9 +/- .22 and 6.78 +/- .47, respectively), compared with the saline group (.03 +/- .02 and 2.55 +/- .27, respectively). The correlation coefficient between mean cuff:vessel and wet:dry ratios was .86 (p = .0016). This study demonstrates that cuff:vessel ratio analysis can be used to identify the distribution of edema fluid versus vessel diameter, and seems to be as effective a technique as gravimetric analysis to quantitate lung water changes in acute lung injury models. Moreover cuff:vessel ratio analysis can differentiate modest changes in pulmonary edema by direct quantitation, an important end-point not provided by wet:dry analysis. Therefore, it may be a more sensitive technique when investigating therapeutic interventions in in vivo models of acute lung injury.

A novel derivatization of phenol after extraction from human serum using perfluorooctanoyl chloride for gas chromatography-mass spectrometric confirmation and quantification.

Phenol (carbolic acid) is widely used as a disinfectant as well as in the chemical industry as an intermediate in the synthesis of a variety of chemicals. Phenol is also the major metabolite of benzene which is used in many commercial solvents. Phenol is toxic and caustic and may cause death even from dermal absorption. Therefore, measurement of phenol in postmortem blood is essential. The concentration of phenol in blood can be measured by gas chromatography with flame ionization or mass spectrometry. Phenol can also be analyzed by high performance liquid chromatography. However, in forensic toxicology, unambiguous confirmation of phenol by mass spectrometry is as important as quantification in blood. Here we describe a novel derivatization of phenol after extraction with chloroform from human serum using perfluorooctanoyl chloride. The perfluorooctanoyl derivative of phenol showed a strong molecular ion at m/z 490 (relative abundance: 23%) whereas the base peak was observed at m/z 77. The derivative of the internal standard 3,4-dimethylphenol showed a very strong molecular ion at m/z 518 (relative abundance: 56%) and the base peak was observed as m/z 121. The derivative of p-cresol, a chemically related phenolic compound, showed a strong molecular ion at 504 m/z (relative abundance: 54%) and a base peak at m/z 107. We observed baseline separation between derivatized phenol (retention time: 6.1 min), p-cresol (retention time: 7.8 min), and the internal standard (retention time: 9.4 min). We observed no interferences in our assay from grossly hemolyzed serum. Within and between run precision was studied using a serum standard containing 25 mg/L of phenol. The within run precision was 6.6% (mean = 24.3, SD = 1.6 mg/L, n = 8) whereas the between run precision was 8.6% (mean = 25.5, SD = 2.2 mg/L, n = 8). The assay was linear for serum phenol concentrations of 10-200 mg/L. The detection limit was 1 mg/L of serum phenol concentration. The average recoveries were 92.1% to 94.0% for various serum phenol concentrations.

Lamotrigine analysis in plasma by gas chromatography-mass spectrometry after conversion to a tert.-butyldimethylsilyl derivative.

Lamotrigine (lamictal) is a new anticonvulsant drug recently approved by the FDA for clinical use. Therapeutic monitoring of lamotrigine is useful for patient management (therapeutic range 1-4 microg/ml). Here we describe a gas chromatography-mass spectrometric identification and quantitation of lamotrigine after extraction from human serum and derivatization. Lamotrigine was extracted from alkaline serum with chloroform and derivatized with N-methyl-N-(tert.-butyldimethysilyl) trifluoroacetamide containing 2% tert.-butyldimethylchlorosilane. Oxazepam-d5 was used as an internal standard. The tert.-butyldimethylsilyl derivative of lamotrigine showed distinct molecular ions at m/z 483 and 485 as well as other peaks at m/z 426, 370 and 334 for unambiguous identification. The base peak was observed at m/z 199. Similarly, the tert.-butyldimethysilyl derivative of oxazepam-d5 showed molecular ions at m/z 519 and 521 along with other characteristic peaks at m/z 462, 376 and 318. For the analysis of lamotrigine, the mass spectrometer was operated in the selective ion monitoring mode. The within-run and between-run precisions were 4.3% (mean=3.01, S.D.=0.13 microg/ml) and 5.1% (mean=2.93, S.D.=0.15 microg/ml), respectively at a serum lamotrigine concentration of 3.0 microg/ml. The within-run and between-run precisions were 8.2% (mean=0.49, S.D.=0.04 microg/ml) and 10.6% (mean=0.47, S.D.=0.05 microg/ml), respectively at a serum lamotrigine concentration of 0.5 microg/ml. The assay was linear for serum lamotrigine concentrations of 0.5-20 microg/ml. The detection limit was 0.25 microg/ml. The assay was free from interferences from common tricyclic antidepressants, benzodiazepines, other common anticonvulsants, salicylate and acetaminophen.

Postmortem lipid levels for the analysis of risk factors of sudden death: usefulness of the Ektachem and Monarch analyzers.

Elevated serum cholesterol, triglyceride, and free fatty acid levels have been identified as risk factors for sudden death from cardiovascular disease and increased risk for myocardial ischemia or arrhythmias; therefore, correlation of antemortem and postmortem lipid levels may be useful in establishing the cause, pathophysiology, or familial risk factors of sudden death. In the present study, antemortem (within 72 h) and postmortem (within 24 h) cholesterol, triglyceride, free fatty acid, and albumin levels were analyzed in seven autopsied hospitalized patients from the University of New Mexico Hospital in Albuquerque, New Mexico. The cholesterol, triglyceride, and albumin levels were measured by dry-slide technology on an Ektachem 700 analyzer, and the free fatty acid levels were measured on a Monarch analyzer with a commercially available kit from Wako Chemical. Postmortem cholesterol levels averaged 13% lower than antemortem levels, postmortem triglyceride levels averaged 38% higher than antemortem levels, postmortem free fatty acid levels averaged 23% lower than antemortem levels, and postmortem albumin levels were essentially unchanged (<0.01% higher) from antemortem levels. Whether the antemortem and postmortem differences in lipid levels were the result of postmortem degradation products, a general phenomenon (such as variable enzyme degradation), or an idiosyncracy of the Ektachem or Monarch systems could not be definitely established. These preliminary results suggest that caution should be exercised when interpreting postmortem cholesterol, triglyceride, and free fatty acid levels analyzed on the Ektachem or Monarch systems.

SK&F 86002, a dual cytokine and eicosanoid inhibitor, attenuates endotoxin-induced cardiopulmonary dysfunction in the pig.

Cytokines and eicosanoids are well documented important mediators of endotoxemia. Bicyclic imidazoles are a novel class of nonsteroidal anti-inflammatory compounds that display unique pharmacological profiles by reducing cytokine production and arachidonic acid metabolism. In this study, we evaluated the ability of the bicyclic imidazole, SK&F 86002, to attenuate endotoxin-induced cardiopulmonary dysfunction. Pigs were randomly assigned to one of four groups: LPS (n = 5), given .5 microgram/kg/h 055:B5 Escherichia coli lipopolysaccharide (LPS) intravenously (i.v.) for 6 h; saline (n = 5); SK&F 86002 (n = 3), given 50 mg/kg SK&F 86002 orally 30 min prior to anesthesia; and SK&F 86002 + LPS (n = 5). Administration of LPS resulted in cardiopulmonary dysfunction characterized by decreased stroke volume and arterial oxygen tension, and increased room air alveolar-arterial oxygen gradient, pulmonary arterial pressure, pulmonary vascular resistance, and peak intratracheal pressure. Additionally, LPS administration was associated with leukopenia and increased pulmonary myeloperoxidase activity. Pretreatment with SK&F 86002 attenuated LPS induced hypotension, hypoxemia and bronchoconstriction and blocked the pulmonary hypertension. SK&F 86002 blocked the LPS-induced increase in myeloperoxidase activity, indicating a reduction in pulmonary neutrophil infiltration, but had no effect on systemic leukopenia. Pretreatment with SK&F 86002 significantly attenuated LPS-induced increases in plasma thromboxane B2 and tumor necrosis factor-alpha. We hypothesize that ameliorating effects of SK&F 86002 in this endotoxin model of cardiopulmonary dysfunction are related to inhibition of cytokine and eicosanoid biosynthesis.

Collision of transitional cell carcinoma and renal cell carcinoma. An immunohistochemical study and review of the literature.

A case characterized by a rare synchronous occurrence of transitional cell carcinoma (TCC) of the renal pelvis and renal cell carcinoma (RCC) in the same kidney is presented. A retrospective analysis of 23 similar cases reported in the English literature over the last 71 years demonstrated a male-to-female ratio of 2:1, an average age of 64.5 years, and a left-to-right-side ratio of 3.2:1. The three most common findings at initial examination were hematuria (90%), flank pain (19%), and flank mass (14%). Moreover, 24% of patients had tumor metastases even at initial examination. Thirty-four percent of patients had bladder neoplasms, and 24% of them had a history of cigarette smoking. There is no tendency toward higher grade of malignancy or specific histologic pattern for TCC and RCC when they occur together in the same kidney. Immunohistochemical studies were used to examine TCC and RCC, with special attention paid to the site of their collision, which displayed multifocal lymphatic permeation. Both TCC and RCC were positive for epithelial membrane antigen (EMA) and cytokeratins identified by monoclonal antibodies CAM-5.2, AE1/AE3, and MAK-6. TCC was focally positive for keratin, detectable by antibody 34 beta E12, but RCC was not. The tumor tissue infiltrating the lymphatics, which seemed to be RCC, demonstrated positive staining for EMA and keratins CAM-5.2, AE1/AE3, and MAK-6 and negative staining for keratin 34 beta E12. Interestingly, the tumor in lymphatics displayed strong staining for carcinoembryonic antigen (CEA) but both TCC and RCC in the vicinity were negative. These findings suggest that keratin 34 beta E12 may play a role in the differential diagnosis between TCC and RCC and that tumor-invading lymphatics may change phenotype, including the neoexpression of CEA.

Anticoagulant effects of sulphonated polyurethanes.

Sulphonated polyurethanes have been shown to have excellent blood contacting properties. In this paper, similar polyurethanes which are water soluble have been investigated to determine their influence on thrombus formation. These polymers were shown to delay clotting times in the following ways: by direct complex formation between the polymer and thrombin; by interference with fibrin polymerization; and by complex interactions between polymer, thrombin, plasma antiproteases and fibrinogen in plasma.

Performance profiles of topical antimicrobials in vitro.

The in-vitro efficacy of commercially available topical antimicrobial products against control strains and those from clinical material are compared with an agar diffusion model. The MICs of the constituent antimicrobial compounds have been determined for the same organisms. Plotting the inhibition zone diameters produced by the topical products against the log10 MICs of their constituent antimicrobial compound(s) gives overall product performance profiles for a range of organisms. These profiles confirm that the formulation of a topical product clearly modifies the response obtained with a specific antimicrobial compound.

The effect of nitroglycerin on response to tracheal intubation. Assessment by radionuclide angiography.

The effect of intravenous (IV) nitroglycerin (NTG) on perioperative myocardial ischemia as detected by single pass radionuclide angiocardiography was studied in 20 patients scheduled for elective coronary artery bypass grafting (CABG). Ten patients, selected at random, received IV NTG 1 microgram.kg-1.min-1 (NTG group) and 10 others, IV saline (control group). Anesthetic induction consisted of midazolam 0.2 mg.kg-1, vecuronium 0.1 mg.kg-1, and 50% N2O in O2. ECG leads I, II, and V5 were monitored for ST segment changes. Single pass radionuclide angiocardiography (RNA) was performed at 5 times: prior to induction, prior to tracheal intubation, and at 1, 3.5, and 6 min following intubation. The presence of new regional wall motion abnormalities (RWMA) was determined from each RNA study as compared with the preinduction measurement. Apart from one patient in the control group who developed a new "v" wave after intubation, there was no evidence of ischemia by pulmonary capillary wedge pressure. No ECG evidence of ischemia was detected in any patient. Despite this, new regional wall motion abnormalities were observed in 3 patients in the control group and 1 patient in the NTG group. Blood pressure and heart rate responses of patients with new RWMA were not significantly different from other patients. The low incidence of ischemia in this population precludes a definitive statement regarding the efficacy of IV NTG, but the lower incidence of RWMA in the NTG group suggests a protective effect.

Cardiac conduction interactions of propranolol and verapamil with halothane in pentobarbitone-anaesthetized dogs.

We have studied the effects of propranolol 0.25 mg kg-1 and verapamil 0.075 mg kg-1 on cardiac conduction and refractoriness in 21 dogs anaesthetized with pentobarbitone 30 mg kg-1 using His bundle electrocardiography and programmed stimulation. After baseline studies under pentobarbitone and halothane (1.3 MAC) anaesthesia, the dogs were allocated randomly to two groups: group 1 received verapamil followed by propranolol; group 2 received propranolol followed by verapamil; the drugs were given in a continuous infusion over 10 min. The atrial-His (AH) interval, the atrioventricular node effective (AVERP), and functional (AVFRP) refractory periods, were prolonged by verapamil in both groups, but not the His-ventricle (HV) interval or the ventricular effective refractory period (VERP). AVFRP and VERP were prolonged by propranolol in both groups. Corrected sinus node recovery times were normal after each drug. Heart rate and the rate required to produce Wenckebach were decreased by each drug. The combination of verapamil and propranolol during halothane anaesthesia in dogs has significant cardiac conduction effects; however, no spontaneous AV block occurred during the study.

D. pteronyssinus-tyrosine adsorbate: biological and clinical properties.

Adsorption of D. pteronyssinus on tyrosine gives a depot formulation with retained immunogenicity. Preliminary clinical evaluation of this material showed improvement in the majority of allergic subjects after receipt of the initial six doses, improvement being continued during subsequent maintenance therapy.

Platelet adhesion and contact activation time tests on HEMA coated cellulose acetate membranes.

Surface modification of cellulose acetate dialysis membranes was carried out by 60Co radiation induced graft copolymerization of the hydrogel, hydroxyethyl methacrylate (HEMA). The degree of grafting was controlled by varying the HEMA monomer concentration in the grafting solution and the radiation dose. A continuous flow platelet adhesion test was designed which allows testing under conditions more closely approximating hemodialysis than other small scale in vitro tests. Platelet adhesion on treated membranes fell substantially with increasing surface HEMA concentration. The presence of HEMA on the membrane surface did not affect the membrane activated clotting times significantly.