Subchronic JP-8 jet fuel exposure enhances vulnerability to noise-induced hearing loss in rats.

Both laboratory and epidemiological studies published over the past two decades have identified the risk of excess hearing loss when specific chemical contaminants are present along with noise. The objective of this study was to evaluate the potency of JP-8 jet fuel to enhance noise-induced hearing loss (NIHL) using inhalation exposure to fuel and simultaneous exposure to either continuous or intermittent noise exposure over a 4-wk exposure period using both male and female Fischer 344 rats. In the initial study, male (n = 5) and female (n = 5) rats received inhalation exposure to JP-8 fuel for 6 h/d, 5 d/wk for 4 wk at concentrations of 200, 750, or 1500 mg/m³. Parallel groups of rats also received nondamaging noise (constant octave band noise at 85 dB(lin)) in combination with the fuel, noise alone (75, 85, or 95 dB), or no exposure to fuel or noise. Significant concentration-related impairment of auditory function measured by distortion product otoacoustic emissions (DPOAE) and compound action potential (CAP) threshold was seen in rats exposed to combined JP-8 plus noise exposure when JP-8 levels of 1500 mg/m³ were presented with trends toward impairment seen with 750 mg/m³ JP-8 + noise. JP-8 alone exerted no significant effect on auditory function. In addition, noise was able to disrupt the DPOAE and increase auditory thresholds only when noise exposure was at 95 dB. In a subsequent study, male (n = 5 per group) and female (n = 5 per group) rats received 1000 mg/m³ JP-8 for 6 h/d, 5 d/wk for 4 wk with and without exposure to 102 dB octave band noise that was present for 15 min out of each hour (total noise duration 90 min). Comparisons were made to rats receiving only noise, and thosereceiving no experimental treatment. Significant impairment of auditory thresholds especially for high-frequency tones was identified in the male rats receiving combined treatment. This study provides a basis for estimating excessive hearing loss under conditions of subchronic JP-8 jet fuel exposure.

The acute and long-term effects of Middle East sand particles on the rat airway following a single intratracheal instillation.

Military personnel deployed in the Middle East have emphasized concerns regarding high levels of dust generated from blowing desert sand and the movement of troops and equipment. Airborne particulate matter levels (PM(10); PM < 10 µm) in the region may exceed 1500 µg/m(3), significantly higher than the military exposure guideline (MEG) of 50 µg/m(3). Increases in PM(10) have been linked to a rise in incidences of asthma, obstructive pulmonary disease, lung cancer, and cardiovascular diseases. Male Sprague-Dawley rats received a single intratracheal (IT) instillation of 1, 5, or 10 mg of Middle East PM(10) collected at a military occupied site in Kuwait, silica (positive control), or titanium dioxide (TiO(2); negative control) suspended in 400 µl sterile saline, or saline alone (vehicle control). Twenty-four hours, 3 d, 7 d and 6 mo postexposure (n = 15/group), organs including lung were evaluated for histopathological changes and for particle contaminants. Bronchoalveolar fluid (BALF) was also analyzed for cellular and biochemical parameters, including cytokines and chemokines. Instillation of silica resulted in early, pronounced, sustained inflammation indicated by significant increases in levels of total protein and neutrophils, and activities of lactate dehydrogenase activity and ß-glucuronidase activity. Lower magnitude and transient changes using the same markers were observed in animals exposed to TiO(2) and Middle East PM(10). The results suggest that for acute exposures, this Middle East PM(10) is a nuisance-type dust with relatively low toxicity. However, since average deployment of military personnel to the Middle East is 180 d with potential for multiple follow-on tours, chronic exposure studies are needed to fully understand the pulmonary effects associated with Middle East PM exposure.

Molecular mechanism of manganese exposure-induced dopaminergic toxicity.

Manganese (Mn) is an essential mineral that is found in varying amounts in aerosols or dust. Exposure to atmospheric Mn at high concentration is a risk factor in humans that can manifest as neuronal degeneration resembling Parkinson's disease (PD). Since the underlying mechanism of Mn and dopamine (DA) interaction-induced cell death remains unclear, here, we showed that Mn exposure alone to mesencephalic cells for 24h induced minimal apoptotic cell death. However, cells pre-exposed to DA for 2h accelerated Mn-induced apoptosis. The vulnerability of Mn-induced apoptotic cell death to DA was determined by measuring lactate dehydrogenase (LDH) and Apoptag TUNEL staining (terminaldeoxynucleotidyl transferase DNA labeling). This was further confirmed by the cell viability assay to support our hypothesis that DA at the cellular level interacts with Mn and causes cells to be more susceptible. Pretreatment with nitric oxide blocker (7-nitroindazole, 7-NI), vitamin E or NF-kappaB inhibitor (SN50) significantly protected the cells from Mn and DA interaction-induced reactive oxygen species (ROS) and apoptosis. Western blot analysis showed that Mn in the presence of DA markedly induced induction of NOS (iNOS) expression. Pretreatment with 7-NI, SN50 or vitamin E significantly attenuated increased iNOS expression indicating that iNOS expression is regulated by ROS and the transcription factor NF-kappaB. Further, the generation of ROS as an early event in Mn and DA interaction is not controlled by NF-kappaB as SN50 pretreatment did not prevent ROS. These findings suggest that NF-kappaB induction and the activation of nitric oxide synthase through ROS represent a proximate mechanism for Mn-induced neurotoxicity.

Prevalence of hantavirus antibody in patients with chronic renal disease in Egypt.

In Egypt, the etiology of chronic renal failure (CRF) is not well defined. A hospital-based case-control study was initiated in February 1998, to determine whether hantavirus infection is involved in chronic renal disease (CRD) in Egypt. The study enrolled 350 study patients with a history of CRF and 695 matched controls with CRD due to renal calculus or renal cancer, but with normal renal functions. Sera from cases and controls were tested for anti-hantavirus IgG using ELISA with a cell-lysate antigen from Hantaan virus prototype strain 76-118. A demographic questionnaire was completed for each study participant. Five of the 350 cases (1.4%), and seven of the 695 controls (1.0%) were antibody-positive to hantavirus, with a titer > or =1:400. The difference in antibody prevalence between the study cases and the control cases was not statistically significant (P = 0.48). All antibody-positive study cases and controls had been exposed to rodents. Data indicated that in Egypt, hantavirus seroprevalence in CRD patients is low, and hantavirus infections do not appear to be a significant cause of CRF.

The changing economics of HIV care.

New advances in human immunodeficiency virus (HIV) monitoring and therapeutics have led to dramatic changes in the course of HIV disease. We evaluated our closed clinic of 425 HIV patients over the period 1995-1998 to determine the cost effectiveness of these changes in care. We found that the costs of antiretroviral therapy tripled over the period of observation, but that these increases were largely offset by major declines in inpatient and home health expenditures. In addition, we found that annual mortality among our HIV patients had declined by 90%. We calculated that the cost per life-year gained is about $17,500, which compares favorably with medical expenditures for renal dialysis or advanced cardiac disease.

Effect of abciximab on cardiac enzyme elevation after transluminal extraction atherectomy (TEC) in high-risk saphenous vein graft lesions: comparison with a historical control group.

Saphenous vein graft (SVG) intervention has been associated with an increased incidence of distal embolization. Long lesions and lesions associated with thrombus are particularly at increased risk. This study was performed to determine whether abciximab may decrease this risk in high risk SVG angioplasty. From June 1994 to June 1998, 84 patients with at least one high risk factor, i.e., lesion length >20 mm or angiographic evidence of thrombus, underwent Transluminal extraction atherectomy (TEC) procedure followed by balloon dilatation or stenting. Of these 84 patients, 37 who had procedure after September 1995 underwent TEC with abciximab (Abciximab Group) and 47 who had their procedure before that date had TEC without abciximab thereby serving as historic control (Non-Abciximab Group). All patients had normal pre-procedure CK and CK-MB. Total creatine kinase (CK) and CK-MB were measured every 8 hr post-procedure for 24 hr. Baseline demographics, angiographic characteristics, incidence of LV dysfunction and triple vessel disease were similar between the two groups. Graft age was similar between two groups (122 +/- 70 vs. 117 +/- 54 months). Graft diameter, pre and post-procedure percent stenoses were not different between the two groups. Stents were used in 65% in the Abciximab group and 45% in Non-Abciximab group (P = 0. 14). There was no in-hospital repeat PTCA, urgent bypass surgery, or cardiac death. There was no difference between the two groups in regards to the incidence of any elevation of total CK (27% vs. 21. 3%) or CK-MB (54% vs. 51%). When used in conjunction with TEC in treating high risk vein graft lesions, abciximab did not reduce post procedure CK-MB elevation in this patient population.

Stent procedure complicated by thrombus formation distal to the lesion within a muscle bridge.

We report here a case of a patient who underwent percutaneous intervention to the left anterior descending artery, complicated by thrombus formation within the myocardial bridge distal to the lesion. There was complete angiographic resolution of thrombus and restoration of the normal antegrade blood flow after infusion of glycoprotein IIb/IIIa antagonist (abciximab). Our observation may suggest that the presence of myocardial bridging distal to coronary lesions should be considered seriously in preprocedural evaluation of the lesions as a potential risk factor for intracoronary thrombus formation. The main coronary arteries and the proximal segments of their major branches lie free on the epicardial surface of the heart. However, in some instances these vessels may penetrate into the muscle being surrounded by the myocardium, with the overlying muscle referred to as a "bridge". Myocardial bridging appears to be a congenital anomaly, due to failure of exteriorization of the primitive coronary intratrabecular arterial network. It occurs in 5-86% of patients in autopsy studies, and it is observed as systolic coronary artery narrowing in 0.5-12% of patients undergoing coronary arteriography. Although the gross anatomist had long recognized that the epicardial coronary artery might on occasion course directly through a segment of cardiac muscle, the physiological significance of this phenomenon was considered benign. This is partly because traditional teaching concerning coronary blood flow delivery to the left ventricular myocardium emphasized the primacy of the diastolic phase of the cardiac cycle. However, myocardial bridging is not always a benign finding, with recent reports suggesting an association with myocardial ischemia, infarction, vasospasm, cardiac arrythmias, and sudden death.

Isolation, sequencing and expression of the superoxide dismutase-encoding gene (sod) of Nocardia asteroides strain GUH-2.

Nocardia asteroides (Na) superoxide dismutase (SOD) has been implicated as a virulence factor that allows the organism to survive intracellular killing by phagocytic cells. A full-length Na sod gene from a pathogenic strain of Na (strain GUH-2) was cloned from a recombinant phage library using the Mycobacterium tuberculosis (Mt) sod gene (Mt sod) as a probe. The promoter region and structural gene (624 bp) of Na sod was sequenced and nucleotide sequence comparisons reveal 77% homology with Mt sod. The Na sod gene also shares considerable sequence homology with sod of other mycobacterial species. In addition, conserved amino acid (aa) sequences important for metal binding indicate that Mn2+ is the preferred metal ion ligand for Na SOD. An Na sod expression plasmid, pYEX1, under transcriptional control of the Mt hsp70 promoter (pY6013), produced a 25-kDa protein product which showed SOD activity when stained in a native polyacrylamide gel and reacted with rabbit polyclonal antibody specific for Na SOD by Western blot. pYEX1, via transformation, was able to complement an Escherichia coli double sodAB mutant deficient in SOD production in the presence of paraquat (methyl viologen) which stimulates the production of superoxide radicals.

Enhancement of liposome-mediated gene transfer into vascular tissue by replication deficient adenovirus.

For both in vitro and in vivo experiments, especially in vascular cells, an efficient and easy method of gene transfer into this tissue would be extremely useful. Previous methods have either yielded low levels of expression or require complicated manipulation of viral vectors. The goal of this study was to develop an easy, efficient method to introduce unmodified plasmid DNA into vascular tissue. In this report it is demonstrated that complexing unmodified plasmid DNA with replication-deficient adenovirus (Ad5 dl312) via cationic lipids enhances gene transfer up to 1000-fold in cultured bovine aortic endothelial cells (BAECs). Further, utilizing a balloon-injured rabbit femoral artery model, intense nuclear staining in both the neointimal smooth muscle cell layer and the adventitia was seen following transfection with a plasmid containing the lacZ gene and the SV40 nuclear localization signal. Control arteries demonstrated no detectable staining. Our studies suggest that complexing plasmid DNA with adenovirus via lipids greatly enhances gene transfer both in vivo and in vitro. This method could have a wide range of applications for experiments in vascular tissue.

Minimizing the risk of inappropriately administering thrombolytic therapy (Thrombolysis and Angioplasty in Myocardial Infarction [TAMI] study group).

Despite the proven benefits of thrombolytic therapy in acute myocardial infarction, concern for its complications, especially in patients misdiagnosed with myocardial infarction, has led to hesitancy in its use. Historical, clinical and electrocardiographic criteria were developed for enrolling patients with suspected acute myocardial infarction into thrombolytic trials by noncardiovascular specialists. The incidence of misdiagnosis of myocardial infarction and the clinical outcomes when these criteria were used were evaluated for 1,387 consecutive patients given thrombolytic therapy. Twenty-five community hospitals and 7 interventional centers were the sites of enrollment. Most patients (63%) were enrolled from community hospitals. Criteria for thrombolytic therapy included: symptoms of acute myocardial infarction < 6 hours but > 20 minutes, and not relieved by nitroglycerin; and ST-segment elevation > or = 1 mm in 2 contiguous leads or ST-segment depression of posterior myocardial infarction. Exclusion criteria reflecting increased risk of bleeding were used. A final diagnosis of myocardial infarction was based on creatinine kinase-MB, electrocardiographic and ventriculographic evaluation. Acute myocardial infarction was misdiagnosed in 20 patients (1.4%; 95% confidence interval 0.8-2.0%). These patients were demographically similar to those with acute myocardial infarction. All misdiagnosed patients survived; no significant adverse events occurred. Thus, in several clinical settings, a simple algorithm with specific criteria was used for diagnosing acute myocardial infarction and administering thrombolytic therapy. The inclusion criteria used in this study led to a low rate of misdiagnosis.

Early and late outcome following deployment of a new flexible tantalum intracoronary stent in dogs.

A new radiopaque, highly flexible balloon-expandable tantalum stent was tested. Thirty-six of 40 stents were successfully deployed percutaneously in the coronary arteries of 31 dogs. The dogs were given aspirin before, intravenous heparin during, and aspirin alone after the procedure. One dog died at 24 hours because of coronary occlusion following traumatic implantation. Four dogs were put to death early, revealing re-endothelialization by 9 days. Eleven dogs were put to death from 2 weeks to 9 months during long-term follow-up, showing all vessels widely patent with the stent uniformly embedded within a stable neointimal layer. Follow-up arteriography showed patency in all remaining stents up to 1 year, with no perforation or aneurysm formation. Four stents were placed into canine peripheral arteries and were removed percutaneously after deployment. Pathology revealed no significant trauma to involved vessels. This tantalum stent exhibits feasibility of percutaneous deployment, early neointimal formation, low thrombogenicity on long-term aspirin therapy alone, and patency up to 1 year in this canine model.

Gene transfer into coronary arteries of intact animals with a percutaneous balloon catheter.

Genetic manipulation of the vasculature may offer insights into the pathogenesis of coronary artery disease and may lead to gene therapy for disorders such as restenosis after percutaneous coronary angioplasty. The goal of this study was to develop a percutaneous method for gene transfer into coronary arteries of intact animals. Liposomes were used to facilitate transfection in coronary arteries with a plasmid containing the cDNA encoding luciferase. This reporter was chosen since it is not expressed in mammalian cells, and it can be quantified using a sensitive assay (light production). Mongrel dogs were catheterized, and DNA was delivered to coronary arteries via a porous perfusion balloon system. Luciferase expression was measured 3-5 days after the procedure, when the dogs were killed. Luciferase activity in control arteries (n = 12) was no higher than average background activity. Eight of 12 transfected arteries exhibited gene expression, averaging 4.3 +/- 2.1 pg luciferase (p less than 0.01, transfected versus control arteries). In addition, the ability to transfect DNA into femoral arteries without a transfection vehicle was tested. Five dogs were subjected to surgical transfection attempts in their femoral arteries with either DNA alone or DNA plus liposomes. Luciferase was expressed in all 10 femoral arteries; those treated with DNA alone expressed 35.6 +/- 8 pg luciferase, and those treated with DNA plus liposomes expressed 42.3 +/- 14 pg luciferase (p = 0.70). These results demonstrate the use of a percutaneous catheter to achieve gene transfer and expression in coronary arteries of intact dogs and suggest that the efficiency of intra-arterial gene transfer may be similar whether or not a transfection vehicle is used.

Recurrent tonsillitis. The role of Chlamydia and Mycoplasma.

The core tissue microflora of 40 patients who underwent tonsillectomies were examined with cultures, DNA probe tests, enzyme immunoassays, and direct immunofluorescence antibody tests for Chlamydia and Mycoplasma. We believe this is the largest and most accurate prospective study that has examined the role of Chlamydia and Mycoplasma in the core tonsil tissue of patients with recurrent or chronic tonsillitis. The data strongly indicate that, unlike acute tonsillopharyngitis, Chlamydia and Mycoplasma are not involved in recurrent or chronic tonsillitis.

Direct in vivo gene transfer into the coronary and peripheral vasculatures of the intact dog.

Gene therapy approaches have been suggested for the treatment of cardiovascular disease. Recently, direct transfer of the gene encoding beta-galactosidase into peripheral arteries of the pig has been demonstrated. To determine whether this approach is applicable to other arterial beds and to other species, we first evaluated the use of beta-galactosidase as a marker protein in the canine model. We demonstrate that variable but substantial endogenous beta-galactosidase-like activity is induced by manipulation of canine peripheral arteries, which precludes the use of this marker protein in evaluating the efficiency of gene transfer in this model. A marker gene encoding firefly luciferase was then evaluated, and background luciferase activity was found to be low in the dog even after arterial manipulation. Using the luciferase gene, we then demonstrated lipid-mediated gene transfer directly into both coronary and peripheral arteries of the intact dog. These results indicate the feasibility of in vivo gene transfer into coronary arteries and demonstrate the use of the luciferase marker protein in quantifying recombinant protein expression following gene transfer in canine models. This simple and effective method for direct in vivo gene transfer into coronary and peripheral arteries may be applicable to the localized production of therapeutically important proteins for the treatment of cardiovascular diseases.

Intravascular stents.

Intravascular stents have been developed to address acute arterial closure and restenosis, the major limitations of percutaneous transluminal coronary angioplasty (PTCA). Metallic stents in human clinical trials have shown efficacy in treating acute closure and, in selected patients, lowering the restenosis rate. This review delineates the characteristics of the ideal stent and examines ongoing clinical trials that are evaluating various stent prototypes. Developmental stents that use radiopaque and bioabsorbable materials are presented. The potential role for intravascular stents as vehicles for localized drug delivery and gene therapy is discussed.

Preservation of resting potential by magnesium in hypoxic canine cardiac cells.

This study was designed to test the hypothesis that Mg++ prevents the electrophysiologic changes caused by hypoxia in cardiac cells. Canine right ventricular false tendons (32 hearts) were pinned to the floor of a tissue bath and suffused with physiologic solution. Microelectrodes recorded the spontaneous action potentials. The isolated tissue was exposed to a hypoxic solution (PO2 less than 50 mm Hg) containing either normal [Mg++] (0.85 mmol/l) or high [Mg++] (3.4 mmol/l). Hypoxia with normal [Mg++] caused increased firing rate (+40%), depolarization of the resting (maximum diastolic) potential (from-72 to -55 mV), and reduction in amplitude (from 88 to 76 mV). The depolarization, reduction in amplitude, and most of the increased firing rate were completely blocked when [Mg++] was elevated to 3.4 mmol/l; propranolol (10 mg/l) blocked the residual increase in firing rate.

Beam pathlength multiplication.

The reflected beam pattern within a beam path multiplier (BPM) formed by two roof reflectors is described in detail. Multiple reflections produced by both translation and rotation of the ridge of one reflector with respect to the other are described in a Cartesian tensor formulation. Equations are derived that describe the spatial distribution of the reflected beams in a BPM of this type under static and dynamic conditions.

Interferometric angular measurement.

An interferometer designed to measure and/or generate angles with high precision and accuracy is described. Design criteria and equations are developed and the experimental performance of a prototype interferometer is discussed.