Collaborative study for the characterization of a chikungunya virus RNA reference reagent for use in nucleic acid testing.

BACKGROUND AND OBJECTIVES: Infections with the mosquito-borne chikungunya virus (CHIKV) can cause febrile illness or be asymptomatic. Laboratory diagnosis of CHIKV is often made with laboratory-developed nucleic acid amplification technology (NAT) assays because there are no U.S. Food and Drug Administration (FDA)-approved diagnostic or blood screening assays. We aimed to produce a well-characterized CHIKV RNA reference reagent (CHIKV-RR) for use in NAT assays. MATERIALS AND METHODS: A CHIKV RNA-RR consisting of cell culture-grown, heat-inactivated CHIKV diluted in human plasma was assessed by 8 laboratories in a collaborative study. The participants were asked to test the CHIKV-RR using their NAT assay(s) by qualitative testing (determination of RNA end-point by testing log and half-log dilutions followed by calculation of estimated NAT-detectable units/ml, after adjustment for the sample volume used for testing), and by quantitative testing, when available. RESULTS: Results from the testing showed that the CHIKV-RR had an estimated overall mean of 7.56 log10 detectable units/ml, ranging from 6.2 log10 to 8.6 log10. CONCLUSIONS: The Center for Biologics for Evaluation and Research/FDA CHIKV RNA-RR for NAT was established with a concentration of 7.56 log10 detectable units/ml.

The adrenocortical response of greater sage grouse (Centrocercus urophasianus) to capture, ACTH injection, and confinement, as measured in fecal samples.

Investigators of wildlife populations often utilize demographic indicators to understand the relationship between habitat characteristics and population viability. Assessments of corticosterone may enable earlier detection of populations at risk of decline because physiological adjustments to habitat disturbance occur before reproductive diminutions. Noninvasive methods to accomplish these assessments are important in species of concern, such as the greater sage grouse (GRSG). Therefore, we validated a radioimmunoassay that measures immunoreactive corticosterone metabolites (ICM) in fecal samples and used it to characterize the adrenocortical response of 15 GRSG exposed to capture, intravenous injection of 50 IU/kg adrenocorticotrophic hormone (ACTH) or saline, and 22 h of confinement. Those animals injected with ACTH exhibited a more sustained (P = 0.0139) and less variable (P = 0.0012) response than those injected with saline, indicating different levels of adrenocortical activity. We also found that potential field-collection protocols of fecal samples did not alter ICM concentrations: samples held at 4 degrees C for up to 16 h contained similar levels of ICM as those frozen (-20 degrees C) immediately. This study demonstrates a multiphasic adrenocortical response that varied with the level of stimulation and indicates that the assay used to measure this phenomenon is applicable for studies of wild GRSG.

Mycobacteriosis-associated mortality in wild striped bass (Morone saxatilis) from Chesapeake Bay, U.S.A.

The striped bass (Morone saxatilis) is an economically and ecologically important finfish species along the Atlantic seaboard of the United States. Recent stock assessments in Chesapeake Bay (U.S.A.) indicate that non-fishing mortality in striped bass has increased since 1999, concomitant with very high (>50%) prevalence of visceral and dermal disease caused by Mycobacterium spp. Current fishery assessment models do not differentiate between disease and other components of non-fishing mortality (e.g., senescence, predation); therefore, disease impact on the striped bass population has not been established. Specific measurement of mortality associated with mycobacteriosis in wild striped bass is complicated because the disease is chronic and mortality is cryptic. Epidemiological models have been developed to estimate disease-associated mortality from cross-sectional prevalence data and have recently been generalized to represent disease processes more realistically. Here, we used this generalized approach to demonstrate disease-associated mortality in striped bass from Chesapeake Bay. To our knowledge this is the first demonstration of cryptic mortality associated with a chronic infectious disease in a wild finfish. This finding has direct implications for management and stock assessment of striped bass, as it demonstrates population-level negative impacts of a chronic disease. Additionally, this research provides a framework by which disease-associated mortality may be specifically addressed within fisheries models for resource management.

Immunization by particle-mediated transfer of the granulocyte-macrophage colony-stimulating factor gene into autologous tumor cells in melanoma or sarcoma patients: report of a phase I/IB study.

The primary objective of this phase I study was to determine the safety of an autologous tumor vaccine given by intradermal injection of lethally irradiated granulocyte-macrophage colony-stimulating factor (GM-CSF) gene-transfected autologous melanoma and sarcoma cells. Secondary objectives included validation of the gene delivery technology (particle-mediated gene transfer), determining the host immune response to the tumor after vaccination, and monitoring patients for evidence of antitumor response. Sixteen patients were treated with either of two different doses of GM-CSF-treated tumor cells. One patient received treatment with both doses of tumor cells. No treatment-related local or systemic toxicity was noted in any patient. Patients administered 100% treated cells (i.e., with a preparation of tumor cells that had all been exposed to GM-CSF DNA transfection) had a more extensive lymphocytic infiltrate at the vaccine site than did patients given 10% treated cells (a preparation of tumor cells in which 10% had been exposed to GM-CSF transfection) or nontreated tumor. The generation of a systemic immune response to autologous tumor by a delayed-type hypersensitivity response to the intradermal placement of nontransfected tumor cells was noted in one patient. One patient had a transient partial response of metastatic tumor sites. The entire procedure, from tumor removal to vaccine placement, was accomplished in less than 6 hr in all patients. Four of 17 patient tumor preparations produced greater than 3.0 ng of GM-CSF per 10(6) cells per 24 hr in vitro. The one patient with greater than 30 ng of GM-CSF per 10(6) cells per 24 hr in vitro had positive DTH, a significant histologic inflammatory response, and clinically stable disease. This technique of gene transfer was safe and feasible, but resulted in clinically relevant levels of gene expression in only a minority of patients.

Induction of cross clade reactive specific antibodies in mice by conjugates of HGP-30 (peptide analog of HIV-1(SF2) p17) and peptide segments of human beta-2-microglobulin or MHC II beta chain.

HGP-30, a 30 amino acid synthetic peptide homologous to a conserved region of HIV-1(SF2) p17 (aa86-115), has previously been shown to elicit both cellular and humoral immune responses when conjugated to KLH and adsorbed to alum. However, the free HGP-30 peptide is not immunogenic in animals. In order to improve the immunogenicity of HGP-30, peptide conjugates consisting of a modified HGP-30 sequence (m-HGP-30/aa82-111) and a peptide segment, residues 38-50, of the MHC I accessory molecule, human beta-2-microglobulin (beta-2-M), referred to as Peptide J, or a peptide from the MHC II beta chain (peptide G) were evaluated in mice. The effects of carriers and adjuvants on serum antibody titers, specificities to various HIV-1 clade peptides similar to HGP-30 and isotype patterns were examined. Peptides J or especially G conjugated to modified-HGP-30 (LEAPS 102 and LEAPS 101, respectively) generated comparable or better immune responses to modified HGP-30 than KLH conjugates as judged by the induction of: (1) similar antibody titers; (2) broader HIV clade antigen binding; and (3) antibody isotype response patterns indicative of a TH1 pathway (i.e. increased amounts of IgG2a and IgG2b antibodies). The ISA 51 and MPL(R)-SE adjuvants induced higher antibody responses than alum, with the ISA 51 being more potent. Immune responses to LEAPS 102, as compared to LEAPS 101, were weaker and slower to develop as determined by antibody titers and cross clade reactivity of the antibodies induced. Compared to KLH conjugates which induced significant anti-KLH antibody titers, minimal antibody responses were observed to peptide G, the more immunogenic conjugate, and peptide J. These results suggest that modified HGP-30 L.E.A.P.S. constructs may be useful as HIV vaccine candidates for preferential induction of TH1 directed cell mediated immune responses.

Analysis of tumor characteristics and survival in liver transplant recipients with incidentally diagnosed hepatocellular carcinoma.

The use of orthotopic liver transplantation (OLTX) for the treatment of hepatocellular carcinoma (HCC) has generally become restricted to carefully selected cases of small oligocentric tumors. However, it is not uncommon to find previously undetected HCC within recipient cirrhotic livers at the time of hepatectomy. The impact of unsuspected HCC on patient outcomes remains unclear. A retrospective analysis of our institutional experience with adult primary OLTX was performed comparing recipients with incidental HCC (group 1), recipients with known or suspected HCC (group 2), and recipients confirmed by pathologic examination to be tumor free (group 3). Between 1984 and 2000, 27 patients in group 1, 12 patients in group 2, and 612 patients in group 3 underwent primary OLTX. Tumors were smaller (P = 0.0172) in group 1 than in group 2; however, the number of tumors and the histologic findings were similar in the groups. Incidence of bilobar involvement, vascular invasion, portal vein tumor thrombus, lymphatic involvement, and distant metastasis at the time of OLTX did not differ significantly between these groups. Four-year patient survival appeared to be lower in group 1 (70.0%) than in group 3 (79.0%) (P = 0.0546); 4-year patient survival was significantly worse in group 2 (31.0%) compared to group 3 (P = 0.0106). Thus, in our experience, incidentally diagnosed cases of HCC possess many of the same features of malignancy as preoperatively diagnosed HCC. Indeed, patient survival after OLTX appears to be adversely affected by the presence of incidental HCC.

Personality and voice disorders: a multitrait-multidisorder analysis.

To examine whether personality has causal, concomitant, or outcome status in common voice disorders, a vocally normal control group and 4 groups with voice disorders--functional dysphonia (FD), vocal nodules (VN), spasmodic dysphonia (SD), and unilateral vocal fold paralysis (UVFP)--were compared using the Multidimensional Personality Questionnaire (MPQ). Univariate and discriminant analyses confirmed that the FD and VN groups differed significantly from each other, the controls, and the other voice-disordered groups at both a superfactor and lower-trait level of personality description. The FD group was introverted, stress reactive, alienated, and unhappy. The VN group was described as socially dominant, stress reactive, aggressive, and impulsive. Comparisons involving the SD, UVFP, and the control subjects did not identify consistent personality differences. While group trends supported the central tenets of a trait theory outlining the dispositional bases of FD and VN, further research is needed to elucidate the specific causal mechanism(s) in individual cases. The disability hypothesis, which suggests that personality features represent the negative effects of vocal disability, was not strongly supported.

Magnetic resonance angiography of aorto-iliac disease.

BACKGROUND: Four different techniques for aorto-iliac magnetic resonance angiography (MRA) were assessed for accuracy using a digital subtraction angiography (DSA) gold standard. Surgeons' confidence in their ability to generate treatment plans with MRA and DSA was assessed, in consultation with a radiologist. METHODS: Two different two-dimensional (2D) time-of-flight (TOF) sequences, a phase-contrast sequence, and a contrast-enhanced (CE) MRA sequence were used. Receiver operating characteristic (ROC) curves were plotted and areas (A(z)) calculated from radiologists' readings. Surgeons' confidence in their ability to utilize the images for treatment planning was assessed with a 5-point Likert scale. Thirty-six patients were evaluated. RESULTS: CE MRA had a sensitivity, specificity, and A(z) of.92,.93, and.96, respectively, for stenoses 50% or greater. CE MRA performed better than other sequences, but the improvement compared with gated 2D TOF was not statistically significant. Interobserver agreement for CE MRA and DSA yielded identical Kappa values. Surgeons were most confident in DSA, followed by CE MRA, which was significantly preferred to other techniques. CONCLUSIONS: CE MRA closely approximates DSA in terms of diagnostic accuracy. Surgeons considering treatment plans are confident in the CE MRA technique, relative to other MRA methods.

Combined hepatocyte-selective and blood-pool contrast agents for the CT detection of experimental liver tumors in rabbits.

PURPOSE: To determine the imaging characteristics of a new computed tomographic (CT) contrast material with both hepatocyte-selective and blood-pool components (iodinated triglyceride (ITG)-dual) versus standard iohexol. MATERIALS AND METHODS: VX2 carcinoma was inoculated in seven rabbits. Animals underwent nonenhanced, iohexol-enhanced (600 mg of iodine per kilogram of body weight), and ITG-dual-enhanced (blood-pool moiety, 100 mg of iodine per kilogram; hepatocyte-selective moiety, 100 or 200 mg of iodine per kilogram, injected 90 minutes apart) helical CT. Livers were removed, preserved in formalin, suspended in agar, and sectioned transversely at 3-mm intervals. Attenuation values for normal liver and tumors were obtained, and blinded readers evaluated images for lesions by using a modified free-response receiver operating characteristic (ROC) method. RESULTS: A total of 47 separate tumor sites were detected at pathologic examination. ITG-dual-enhanced scans obtained with 300 mg of iodine per kilogram demonstrated similar liver opacification to iohexol-enhanced scans obtained with 600 mg of iodine per kilogram, but with less lesion enhancement, which resulted in better liver-to-lesion contrast. Blinded readers had a higher sensitivity, accuracy, and area under the ROC curve for ITG-dual-enhanced scans as compared with iohexol-enhanced scans (P: <.01). CONCLUSION: ITG-dual-enhanced CT quantitatively and qualitatively improved liver lesion detection versus iohexol-enhanced CT. Future clinical trials with various human tumor types after potential approval for human use are needed to determine the ultimate role of this or other dual-mechanism contrast materials.

Acute renal failure requiring hemodialysis immediately after heart transplantation portends a poor outcome.

BACKGROUND: Previous studies have not provided a definite clarification for the predictive value of pretransplant renal indices on postcardiac transplant patient outcome. Therefore, the purpose of this study was to investigate the interaction between pretransplant renal function and recovery after heart transplantation. METHODS: The study group consisted of 199 consecutive patients who underwent heart transplantation between 1973 and 1994. For better comparison, patients were arbitrarily divided into three different groups based on the year of the transplant operation: Group I- before 1985 (n=13), Group II- between 1985 and 1989 (n=68) and Group III- between 1990 and 1994 (n=118). Values for serum creatinine (Cr), blood urea nitrogen (BUN), urea/creatinine ratio (U/Cr), creatinine clearance (Cr(cl)), length of hospital stay (LOS), early (30-day) mortality, and survival at 1-year and at 5-year were collected for each patient. The data was analyzed by the use of univariate log-rank test with forward stepwise procedure. RESULTS: Postcardiac transplant LOS in the hospital or survival was unaffected by the pretransplant renal indices except the U/Cr ratio (p>0.05). When adjusted for the time, the U/Cr ratio was also insignificant (p=0.1349). The use of hemodialysis was necessary in 9 patients (4.5%) for treatment of acute renal failure manifested immediately after the transplant operation. Early mortality was 44% for these 9 cardiac transplant recipients who required the use of hemodialysis: 0% (0/3) in the 1985-1989 period and 67% (4/6) in the 1990-1994 period. CONCLUSIONS: Pretransplant renal indices have no predictive value on outcome after a heart transplant operation, however, postcardiac transplant acute renal failure necessitating hemodialysis portends a poor outcome.

Personality and voice disorders: a superfactor trait analysis.

To determine whether personality factors play causal, concomitant, or consequential roles in common voice disorders, a vocally normal control group and four groups with voice disorders-functional dysphonia (FD), vocal nodules (VN), spasmodic dysphonia (SD), and unilateral vocal fold paralysis (UVFP)-were compared on measures of personality and psychological adjustment. Superfactor group comparisons revealed that the majority of FD and VN subjects were classified as introverts and extraverts, respectively. Comparisons involving the SD, UVFP, and control subjects did not identify consistent personality differences. The disability hypothesis, which suggests that personality features and emotional maladjustment are solely a negative consequence of vocal disability, was not supported. Personality variables and their behavioral consequences may therefore contribute to FD and VN. Results are presented within the context of a dispositional theory offered by Roy and Bless (2000a).

Determination of stone composition by noncontrast spiral computed tomography in the clinical setting.

OBJECTIVES: Several investigators have evaluated noncontrast computed tomography (NCCT) in predicting stone composition in vitro. We assessed NCCT in predicting stone composition in patients presenting to our emergency room with flank pain and stone disease. METHODS: One hundred twenty-nine patients presenting to our university hospital with flank pain underwent renal colic protocol NCCT scans at the request of the emergency room physicians. A General Electric, high-speed advantage CT scanner was used at 120 kV, 200 mA, and 1.4:1 pitch, with collimation varying between 3 and 5 mm. Ninety-nine patients with predominantly (greater than 50%) calcium oxalate or uric acid composition after either stone passage or stone removal were identified. Each scan was analyzed by one of two radiologists, who determined the predominant attenuation for each stone. Stones once passed or retrieved were analyzed by Urocor Laboratories. The attenuation and attenuation/size ratio (peak attenuation/size in millimeters) were compared with the results of the stone analysis. RESULTS: Eighty-two calculi predominantly composed of calcium oxalate and 17 calculi predominantly composed of uric acid were identified in 99 patients. The calculi ranged in size from 1 to 28 mm. A significant difference (P = 0.017, unpaired t test) was found between the Hounsfield measurement of uric acid calculi (mean 344 +/- 152 HU) and the Hounsfield measurement of calcium oxalate calculi (mean 652 +/- 490 HU). If only the Hounsfield units from stones 4 mm or larger were compared, the data were even more compelling (P = 0.002). However, using an attenuation/size ratio cutoff of greater than 80, the negative predictive value was 99% that a stone would be predominantly calcium oxalate. CONCLUSIONS: Using peak attenuation measurements and the attenuation/size ratio of urinary calculi from NCCT, we were able to differentiate between uric acid and calcium oxalate stones.

Simultaneous pancreas-kidney transplantation reduces excess mortality in type 1 diabetic patients with end-stage renal disease.

BACKGROUND: Diabetic renal disease continues to be the most significant cause of end-stage renal disease (ESRD) in the United States. Renal transplantation improves diabetic ESRD patient survival; however, the diabetic state remains associated with poor patient survival. Simultaneous pancreas-kidney (SPK) transplantation can restore normoglycemia and thus may improve outcomes. METHODS: We assessed the impact of SPK on age-range-matched type 1 diabetic patients who underwent renal transplantation at a single center. The observed/expected life span and annual mortality rates (AMRs) were used as measures of survival. A Cox proportional hazards analysis was used to analyze the impact of potential variables on mortality in SPK recipients. RESULTS: SPK transplantation (N = 335) increased the observed/expected life span compared with diabetic cadaveric (DM-Cad, N = 147) and live-donor (DM-Live, N = 160) transplant recipients (P = 0.004) and significantly reduced the AMRs (SPK, 1. 5%; DM-Cad, 6.27%; DM-Live, 3.65%, P = 0.008, SPK vs. other DM). Moreover, the SPK observed/expected life span and AMR were not significantly different from that of age-range-matched nondiabetic transplant recipients (N = 492). The only variable that was significantly associated with patient survival was discharge serum creatinine (relative risk 1.16, P < or = 0.0154). CONCLUSION: These data demonstrate that SPK improves the ability for type 1 diabetic patients to live more of their expected life span. This suggests that glycemic control, even as a late intervention in a diabetic patient's lifetime, may beneficially affect survival.

Renin-angiotensin system gene expression in post-transplant hypertension predicts allograft function.

BACKGROUND: Registry analyses and single-center studies have demonstrated that hypertension significantly increases the risk for chronic graft loss. The graft itself may contribute to posttransplant hypertension, and intragraft vasoactive hormones therefore, may be dysregulated in posttransplant hypertension. METHODS: We used the reverse-transcription polymerase chain reaction to assess the intragraft regulation of renin-angiotensin system transcripts in biopsy samples from 42 stable renal transplant patients with posttransplant hypertension. We also examined mRNA expression of inducible nitric oxide synthase, transforming growth factor-beta (TGF-beta), select cytokines, and metalloproteinase transcripts in biopsy tissue. Polymerase chain reaction products were quantitated using high performance liquid chromatography and normalized to beta-actin mRNA expression. Serum creatinine, glomerular filtration rate or creatinine clearance and tubular atrophy on biopsy were concurrently assessed. RESULTS: Renin and select Thl cytokine mRNA expression correlated with blood pressure. Type 1 angiotensin II receptor mRNA expression significantly correlated with glomerular filtration rate or creatinine clearance (P = 0.034) and inversely correlated with Th1 cytokines, inducible nitric oxide synthase, and cyclooxygenase-1 mRNA expression (P< or =0.013 for each). Type 1 angiotensin II receptor mRNA also approached a significant inverse correlation with TGF-beta mRNA expression (P = 0.09). Conversely, angiotensin-converting enzyme mRNA expression directly correlated with Thl cytokine (P< or =0.008 for each) and TGF-beta mRNA expression (P = 0.006). Type 1 angiotensin II receptor mRNA expression also correlated with matrix metalloproteinase-1 promoter region, tissue inhibitor of matrix metalloproteinase-2 (TIMP-2) and tissue inhibitor of matrix metalloproteinase-3 mRNA expression. Notably, matrix metalloproteinase-1 promoter region, tissue inhibitor of matrix metalloproteinase-2, and tissue inhibitor of matrix metalloproteinase-3 inversely correlated with TGF-beta mRNA expression (P< or =0.0027 for each). Type 1 angiotensin II receptor mRNA expression at biopsy directly correlated with glomerular filtration rate at 2 year's follow-up. However, angiotensin-converting enzyme mRNA expression at biopsy inversely correlated with glomerular filtration rate at 2 year's follow-up. CONCLUSIONS: These data suggest that allograft-level RAS gene expression may be predictive of future graft function in the setting of diastolic hypertension.

Simultaneous pancreas-kidney transplantation and living related donor renal transplantation in patients with diabetes: is there a difference in survival?

OBJECTIVE: To compare the outcome of simultaneous pancreas-kidney transplantation (SPK) and living related donor renal transplantation (LRD) in patients with diabetes. SUMMARY BACKGROUND DATA: It remains unanswered whether diabetic patients with end-stage renal failure are better served by LRD or SPK. METHODS: Using a longitudinal database, data from all diabetic patients receiving LRD or cadaveric renal transplants or SPKs from January 1986 through January 1996 were analyzed. Patient and graft survival, early graft function, and the cause of patient and graft loss were compared for 43 HLA-identical LRDs, 87 haplotype-identical LRDs, 379 SPKs, and 296 cadaveric renal transplants. RESULTS: The demographic composition of the SPK and LRD groups were similar, but because of less strict selection criteria in the cadaveric transplant group, patients were 10 years older, more patients received dialysis, and patients had been receiving dialysis longer before transplantation. Patient survival was similar for the SPK and LRD groups but was significantly lower for the cadaveric renal transplant group. Similarly, there was no difference in graft survival between SPK and LRD recipients, but it was significantly lower for recipients in the cadaveric renal transplant group. Delayed graft function was significantly more common in the cadaveric renal transplant group. Discharge creatinine, the strongest predictor of patient and graft survival, was highest in the SPK group and lowest in the HLA-identical LRD group. The rate of rejection within the first year was greatest in SPK patients (77%), intermediate in the haplotype-identical LRD and cadaveric transplant groups (57% and 48%, respectively), and lowest (16%) in the HLA-identical LRD group. Cardiovascular disease was the primary cause of death for all groups. Acute rejection, chronic rejection, and death with a functioning graft were the predominant causes of graft loss. CONCLUSIONS: This study demonstrates that there was no difference in patient or graft survival in diabetic patients receiving LRD or SPK transplants. However, graft and patient survival rates in diabetic recipients of cadaveric renal transplants were significantly lower than in the other groups.

A time-tradeoff method for cost-effectiveness models applied to radiology.

PURPOSE: The wait tradeoff (WTO) is a simple time-tradeoff method designed for temporary health states that uses a realistic and intuitive interface for the patient/subject. This method was tested by assessing patients' preferences for magnetic resonance angiography (MRA) versus x-ray angiography (XRA). MATERIALS AND METHODS: The WTO was tested by telephone interview in 38 patients with atherosclerotic peripheral vascular disease, all having previously undergone both MRA and XRA. At indifference point, patients were ambivalent about having MRA or XRA and immediate treatment, versus having a waiting period for test results and treatment after a hypothetical "ideal test" that entailed no pain or risk. RESULTS: The patients were willing to wait a mean of 42.1 days after the ideal test for results and treatment, as opposed to XRA. They were willing to wait only 16.1 days as opposed to MRA. This difference in waiting times was significant (p = 0.0001) and indicates a clear preference for MRA, in agreement with known literature. CONCLUSION: The WTO method assesses preferences for these radiologic tests in an intuitive fashion that does not invoke artificial or irrelevant health states. This approach may also prove useful for other testing situations or short-term treatments being evaluated for cost-effectiveness.

Clinically stable human renal allografts contain histological and RNA-based findings that correlate with deteriorating graft function.

BACKGROUND: Chronic rejection (CR) remains idiopathic, difficult to prospectively identify, and once detected, unresponsive to increased immunosuppression. We hypothesized that clinically stable human renal allografts have ongoing evidence of injury and immune activity, and that this correlates with the worsening of allograft function characteristic of CR. METHODS: The allografts of 40 stable renal allograft recipients were biopsied 2-3 years after transplantation. Biopsies were processed for histology and RNA extraction. RNA was evaluated by semi-quantitative RT-polymerase chain reaction for CD3y mRNA (a marker of T cell receptor turnover), and mRNA from cytokine genes previously shown to be transcribed during acute rejection: tumor necrosis factor-alpha, interferon-gamma, interleukin- (IL) 1beta, IL-2, IL-4, IL-6, and IL-8. Clinical parameters including urine protein and glomerular filtration rate were measured the day of biopsy. Findings were then compared with clinical outcome to establish associations between subclinical inflammation and graft dysfunction. Allograft function was measured again 2 years after biopsy and correlated with findings at the time of biopsy. RESULTS: Cytokine transcripts and histological evidence of injury were detected in more than two-thirds of stable grafts. The degree of the lymphocytic infiltrate correlated with the degree of proteinuria (P=0.034) and histological fibrosis (P=0.005). Similarly, the degree of intragraft CD3y transcription correlated with increasing proteinuria (P=0.043). IL-6 and IL-8 transcripts were also correlated with evidence of graft injury. After 2 years, those biopsies originally found to have evidence of fibrosis, tubular atrophy, or CD3gamma transcription had worsening graft function as determined by creatinine and glomerular filtration rate. CONCLUSIONS: These data demonstrate that significant injury and immune activity can be detected in patients who are stable on clinical grounds. Undetected subclinical graft injury may be a cause of chronic allograft rejection.

Comparison of treatment plans for lower extremity arterial occlusive disease made with electrocardiography-triggered two-dimensional time-of-flight magnetic resonance angiography and digital subtraction angiography.

BACKGROUND: The purpose of the study was to determine whether preoperative treatment plans for patients with lower extremity ischemia can be made with electrocardiography (EKG)-triggered two-dimensional (2D) time-of-flight (TOF) magnetic resonance angiography (MRA) as accurately as digital subtraction angiography (DSA). METHODS: Forty patients were prospectively evaluated with the combination of EKG-triggered 2D TOF MRA, DSA, and pulse volume recordings. Blinded reviewers graded arterial segments for disease severity. Accuracy of separate MRA- and DSA-based treatment plans was compared with the procedures performed based on all available information. RESULTS: There was an 86% exact match between MRA- and DSA-based plans (92% MRA and 94% DSA accuracy). The MRA-based plan accurately predicted 90% of suprainguinal and 95% of infrainguinal procedures, whereas the DSA-based plan accurately predicted 100% of suprainguinal and 85% of infrainguinal procedures. Two-year primary patency was 83% for all procedures. Radiologists' review of disease severity resulted in a mean exact correlation between studies of 81% (kappa = 0.64). The agreement between radiologists interpreting the MRA was 84% (kappa = 0.7) compared with 82% (kappa = 0.66) for the DSA. CONCLUSIONS: MRA- and DSA-based preoperative management plans were of comparable efficacy. Significant interobserver variability was seen with the interpretations of both preoperative studies. EKG-triggered 2D TOF MRA can be used to plan arterial reconstructions; however, all patients require arterial pressure measurements prior to suprainguinal repair and confirmatory intraoperative angiography during infrainguinal revascularization.

Peripheral vascular disease and renal transplant artery stenosis: a reappraisal of transplant renovascular disease.

BACKGROUND: Renal transplant artery stenosis (RTAS) continues to be a problematic, but potentially correctable, cause of post-transplant hypertension and graft dysfunction. Older transplant recipients, prone to peripheral vascular disease (PVD), may have pseudoRTAS with PVD involving their iliac system. METHODS: We retrospectively analyzed 819 patients who underwent kidney transplantation between 1993 and 1997 to determine the contribution of pseudoRTAS to renal transplant renovascular disease. Univariate analyses were performed for donor and recipient variables, including age, weight, gender, race, renal disease, cholesterol and creatinine values, human leukocyte antigen (HLA) matching, cytomegalovirus (CMV) infection, and immunosuppressive medications. Significant variables were then analyzed by a Cox proportional hazards model. RESULTS: Ninety-two patients (11.2%) underwent renal transplant arteriogram (Agram) or magnetic resonance angiography (MRA) for suspected RTAS. RTAS or pseudoRTAS, defined as one or more hemodynamically significant lesions in the transplant artery or iliac system, was evident in 44 patients (5.4%). Variables significantly associated with RTAS by univariate analysis were weight at the time of transplant (p = 0.0258), male gender (p = 0.034), discharge serum creatinine > 2 mg/dL (p = 0.0041), and donor age (p = 0.0062). Variables significantly associated with pseudoRTAS by univariate analysis were weight at the time of transplant (p = 0.0285), recipient age (p = 0.0049), insulin-dependent diabetes mellitus (IDDM; p = 0.0042), panel reactive antibody (PRA) at transplant (p = 0.018), and body mass index (p = 0.04). Weight at transplant and donor age remained significantly associated with an increased risk for RTAS in a multivariate stepwise Cox proportional hazards model. IDDM, transplant PRA, weight at transplant, and donor age were significantly associated with an increased risk for pseudoRTAS in a multivariate stepwise Cox proportional hazards model. Importantly, both RTAS and pseudoRTAS were associated with poorer graft survival (p < 0.007 for each). CONCLUSIONS: Renal transplant renovascular disease encompasses pre-existing PVD acting as pseudoRTAS, as well as classical RTAS. Efforts to identify and correct renal transplant renovascular disease of either nature are important, given its negative impact on graft survival.