Inhibition of mTOR affects protein stability of OGT.

Autophagy regulates cellular homeostasis through degradation of aged or damaged subcellular organelles and components. Interestingly, autophagy-deficient beta cells, for example Atg7-mutant mice, exhibited hypoinsulinemia and hyperglycemia. Also, autophagy response is diminished in heart of diabetic mice. These results implied that autophagy and diabetes are closely connected and affect each other. Although protein O-GlcNAcylation is up-regulated in hyperglycemia and diabetes, and O-GlcNAcylated proteins play an important role in metabolism and nutrient sensing, little is known whether autophagy affects O-GlcNAc modification and vice versa. In this study, we suppressed the action of mTOR by treatment of mTOR catalytic inhibitors (PP242 and Torin1) to induce autophagic flux. Results showed a decrease in global O-GlcNAcylation, which is due to decreased OGT protein and increased OGA protein. Interestingly, knockdown of ATG genes or blocking of lysosomal degradation enhanced protein stability of OGT. In addition, when proteasomal inhibitor was treated together with mTOR inhibitor, protein level of OGT almost recovered to control level. These data suggest that mTOR inhibition is a more efficient way to reduce protein level of OGT rather than that of CHX treatment. We also showed that not only proteasomal degradation regulated OGT stability but autophagic degradation also affected OGT stability in part. We concluded that mTOR signaling regulates protein O-GlcNAc modification through adjustment of OGT stability.

Adenosine A2A receptor polymorphisms in Korean patients with systemic sclerosis.

Adenosine A2A receptor (ADORA2A) regulates inflammation, promotes tissue repair and collagen production by human dermal fibroblasts. We investigated the genetic polymorphisms of ADORA2A in susceptibility to systemic sclerosis (SSc). We genotyped 142 Korean SSc patients and 150 controls for polymorphisms of -1751A/C (rs5996696) and 1976C/T (rs5751876), to cover the promoter and all exon sequences of ADORA2A in Koreans, using TaqMan fluorogenic 5' nuclease assay and single base primer extension assay. Neither -1751A/C nor 1976C/T polymorphism showed difference in the distribution of alleles or genotypes between patients and controls with allele frequency of 89.9% v 91.0% for -1751A (p=0.64) and 56.5% v 54.0% for 1976C (p=0.55). Our findings suggest that the role of ADORA2A in SSc may not be genetically related.

Monoclonal antibody to murine PECAM-1 (CD31) blocks acute inflammation in vivo.

A murine model of peritonitis was used to test the role of platelet/endothelial cell adhesion molecule 1 (PECAM-1/CD31) in acute inflammation. A monoclonal antibody (mAb) specific for murine PECAM-1 injected intravenously 4 h before the intraperitoneal injection of thioglycollate broth blocked leukocyte emigration into the peritoneal cavity for up to 48 h. This block was particularly evident for neutrophils. Control mAb, including one that bound to murine CD18 without blocking its function, failed to block emigration when used at the same or higher concentrations. The decreased emigration seen with the anti-PECAM-1 antibody was not due to neutropenia or neutrophil sequestration in the lung, spleen, or other organs; peripheral blood leukocyte counts were not diminished in these mice. In the mesenteric venules of the mice treated with anti-PECAM-1 mAb, leukocytes were frequently seen in association with the luminal surface of the vessel, but did not appear to emigrate. Thus, the requirement for PECAM-1 in the transendothelial migration of leukocytes previously seen in an in vitro model holds true in this in vivo model of acute inflammation.

Genetic polymorphisms of the beta 2-adrenergic receptor in nocturnal and nonnocturnal asthma. Evidence that Gly16 correlates with the nocturnal phenotype.

Nocturnal asthma represents a unique subset of patients with asthma who experience worsening symptoms and airflow obstruction at night. The basis for this phenotype of asthma is not known, but beta 2-adrenergic receptors (beta 2AR) are known to downregulate overnight in nocturnal asthmatics but not normal subjects or nonnocturnal asthmatics. We have recently delineated three polymorphic loci within the coding block of the beta 2AR which alter amino acids at positions 16, 27, and 164 and impart specific biochemical and pharmacologic phenotypes to the receptor. In site-directed mutagenesis/recombinant expression studies we have found that glycine at position 16 (Gly16) imparts an accelerated agonist-promoted downregulation of beta 2AR as compared to arginine at this position (Arg16). We hypothesized that Gly16 might be overrepresented in nocturnal asthmatics and thus determined the beta 2AR genotypes of two well-defined asthmatic cohorts: 23 nocturnal asthmatics with 34 +/- 2% nocturnal depression of peak expiratory flow rates, and 22 nonnocturnal asthmatics with virtually no such depression (2.3 +/- 0.8%). The frequency of the Gly16 allele was 80.4% in the nocturnal group as compared to 52.2% in the nonnocturnal group, while the Arg16 allele was present in 19.6 and 47.8%, respectively. This overrepresentation of the Gly16 allele in nocturnal asthma was significant at P = 0.007 with an odds ratio of having nocturnal asthma and the Gly16 polymorphism being 3.8. Comparisons of the two cohorts as to homozygosity for Gly16, homozygosity for Arg16, or heterozygosity were also consistent with segregation of Gly16 with nocturnal asthma. There was no difference in the frequency of polymorphisms at loci 27 (Gln27 or Glu27) and 164 (Thr164 or Ile164) between the two groups. Thus the Gly16 polymorphism of the beta 2AR, which imparts an enhanced downregulation of receptor number, is overrepresented in nocturnal asthma and appears to be an important genetic factor in the expression of this asthmatic phenotype.

Urothelium-specific expression of an oncogene in transgenic mice induced the formation of carcinoma in situ and invasive transitional cell carcinoma.

Although many genetic alterations are known to be associated with human transitional cell carcinoma (TCC) of the urinary bladder, relatively little is known about the roles of these molecular defects, singular or in combination, in bladder tumorigenesis. We have developed a transgenic mouse model of bladder tumorigenesis using a 3.6-kb promoter of uroplakin II gene to drive the urotheliums-specific expression of oncogenes. In this study, we demonstrate that transgenic mice bearing a low copy number of SV40T transgene developed bladder carcinoma in situ (CIS), whereas those bearing high copies developed CIS as well as invasive and metastatic TCCs. These results indicate that the SV40T inactivation of p53 and retinoblastoma gene products, defects frequently found in human bladder CIS and invasive TCCs, can cause the aggressive form of TCC. Our results also provide experimental proof that CIS is a precursor of invasive TCCs, thus supporting the concept of two distinct pathways of bladder tumorigenesis (papillary versus CIS/invasive TCC). This transgenic system can be used for the systematic dissection of the roles of individual or combinations of specific molecular events in bladder tumorigenesis.

Immune reconstitution inflammatory syndrome in a HIV-infected patient with disseminated tuberculosis.

Immune reconstitution inflammatory syndrome (IRIS) is the paradoxical worsening of pre-existing infectious processes after commencement of anti-retroviral therapy (ART) in HIV-infected patients. Its manifestations are dependent on the underlying opportunistic infections. We report a case of an HIV-infected patient with disseminated tuberculosis, who responded to anti-tuberculosis therapy but suffered from paradoxical worsening after commencement of ART.

Role of Ha-ras activation in superficial papillary pathway of urothelial tumor formation.

Urothelial tumors develop along two distinctive phenotypic pathways (superficial papillary non-invasive tumors versus flat carcinoma in situ lesions), with markedly different biological behavior and prognosis. Although multiple genetic alterations have been identified in human bladder cancer, their cause-effect relationship with the two pathways has not been firmly established. Using a urothelium-specific promoter of the uroplakin II gene, we showed earlier in transgenic mice that the urothelial expression of SV40T antigen, which inactivates p53 and pRb, induced carcinoma in situ and invasive and metastatic bladder cancer. In striking contrast, we demonstrate here that the urothelial expression of an activated Ha-ras in transgenic mice caused urothelial hyperplasia and superficial papillary non-invasive bladder tumors. These results provide strong, direct experimental evidence that the two phenotypical pathways of bladder tumorigenesis are caused by distinctive genetic defects. Our results indicate that Ha-ras activation can induce urothelial proliferation in vivo; and that urothelial hyperplasia is a precursor of low-grade, superficial papillary bladder tumors. Our transgenic models provide unique opportunities to study the detailed molecular events underlying different types of bladder neoplasms, and can serve as useful preclinical models for evaluating the in vivo efficacy of preventive and therapeutic agents that act on various signaling pathways in bladder cancer.

Characterization of a mutant T-cell hybridoma line with defects in the TCR-mediated apoptotic pathway.

A mutant T-cell hybridoma line named mutant 51 was developed that, unlike the parental line, did not die after T-cell receptor (TCR) engagement and demonstrated reduced death in response to dexamethasone. Intracellular calcium measurements showed that available calcium stores were markedly reduced in the mutant cell line. Unlike control cells, secretion of IL-2 from mutant cells was also greatly reduced, although addition of exogenous IL-2 did not facilitate increased apoptosis. Although levels of the cell death gene product Nur77 were equivalent, additional studies showed that mutant cells expressed Nur77 predominantly in the cytoplasm following TCR engagement, while parental cells displayed a nuclear translocalization of Nur77. In addition, Fas levels and Fas ligand dependant killing were both markedly reduced in the mutant clone. From these data we hypothesize a role for available calcium stores and Nur77 nuclear localization in TCR-mediated apoptosis in T-cell hybridomas.

Note: advancement in tip etching for preparation of tunable size scanning tunneling microscopy tips.

The two aspects of a scanning tunneling microscopy tip, the macroscopic profile and the nanoscale apex, can be tailored by controlling the tension during electrochemical etching and the solution-electrode contact area via acetone vapor. The apex diameter is shown to be proportional to the square root of the tension, and is demonstrated over apex diameters of 150-500 nm. The apex was found to be created in four distinct shapes where a secondary etching can reshape the tip into a single geometry. Improvement in tip height and stability of the profile are demonstrated versus a non-acetone fabrication control.

The s29x gene of symbiotic bacteria in Amoeba proteus with a novel promoter.

Gram-symbiotic bacteria (called X-bacteria), present in the xD strain of Amoeba proteus as required cell components, synthesize and export a large amount of a 29-kDa protein, S29x. S29x is exported into the host's cytoplasm across the bacterial membranes and the symbiosome membrane. The complete nucleotide (nt) sequence of the s29x gene of X-bacteria has been determined, and the promoter sequence and tsp have also been identified. The gene has a nonconventional promoter with putative nt sequences different from the known consensus sequences. When Escherichia coli cells are transformed with s29x, the gene is expressed and the product is secreted into the culture medium. Functions of S29x are not fully known, but it is suspected that S29x plays an important role in the symbiotic relationship between amoebae and X-bacteria.

Construction and characterization of the soybean leaf metalloproteinase cDNA.

The cloning and analysis of a cDNA clone encoding the soybean metalloproteinase obtained by polymerase chain reaction (PCR) and the rapid amplification of cDNA ends (RACE) reaction are described. The cDNA was constructed from poly(A)+ RNA isolated from 15-17 day old leaves. The deduced amino acid sequence of the cDNA reveals that the plant metalloproteinase is synthesized as a preproenzyme and the proenzyme form shares a structural motif, responsible for maintenance of inactive zymogen, with the matrix metalloproteinase (e.g. collagenase) family of enzymes from vertebrate origin. Northern and Western blot analysis demonstrated that the metalloproteinase transcript and protein are under a strict developmental program in that both are expressed only in leaf tissue and in a temporal fashion. The physiological function of the metalloproteinase still remains unclear although the data suggest that the enzyme is extracellular and a portion of the mature form of the enzyme is tightly bound to the cell wall.

Indications for implantation of a dual-chamber pacemaker combined with an implantable cardioverter-defibrillator.

Of 122 patients with single-chamber implantable cardioverter-defibrillators (ICDs) reviewed retrospectively, 35 had traditional indications, 14 had other indications, and 18 had ICD-specific indications for dual-chamber pacing therapy. Thus, 67 patients (55%) were potential candidates for dual-chamber pacing, which has only recently become available combined with ICD therapy.

Mechanism of bile salt vasoactivity: dependence on calcium channels in vascular smooth muscle.

1. The vasoactive mechanisms of bile salts have been investigated in rat isolated portal venous and superior mesenteric arterial rings and perfused mesentery. 2. The isolated perfused mesentery was precontracted with a selective alpha 1-adrenoceptor agonist, cirazoline. Incremental doses of tauroursodeoxycholate (TUDC), taurochenodeoxycholate (TCDC) and taurodeoxycholate (TDC) caused a dose-dependent vasorelaxation. The order of potency of the vasodilator effect was TDC > TCDC > TUDC. 3. The effect of TDC (1.9 x 10(-8)-1.9 x 10(-6) mol) was examined before and after propranolol (3 microM), tetraethylammonium (5 mM), ouabain (10(-5) M), NG-nitro-L-arginine methyl ester (10(-4) M) and capsaicin (50 mg kg-1) to block, respectively, beta-adrenoceptors, K+ -channels, Na+, K+-ATPase, nitric oxide synthase, and primary sensory nerves. The vasodilator effect of TDC was not affected by any of these blocking agents or by denuding vascular endothelium with distilled water. 4. Infusion of TDC (1.9 x 10(-8)-1.9 x 10(-6) mol) with K+-free or high K+ (60 mM) physiological salt solution (PSS) did not affect the vasodilator effect of TDC. 5. Contractions induced by KCl (0.01-1.0 M), arginine vasopressin (AVP, 10(-10)-10(-7) M) or cirazoline (10(-7) x 10(-5) M) were all inhibited by TDC (300 microM). 6. TDC (10(-6) to 10(-3) M) also inhibited the basal tension and the development of spontaneous contractions in the isolated portal vein. 7. TDC (300 microM), however, did not affect noradrenaline-induced phasic contractions elicited in Ca(2+)-free PSS by Ca2+ release from intracellular stores. 8. We conclude that TDC inhibits Ca2+ entry through both voltage-operated and receptor-operated calcium channels, whereas intracellular Ca2+ release is not affected.

Nasal CPAP in nonapneic nocturnal asthma.

Nasal CPAP has been shown to improve nocturnal asthma in those patients with associated sleep apnea. We studied seven nonapneic, nonsnoring asthmatics to determine the effect of CPAP in this patient population. On the CPAP night vs the baseline night, there was a significant worsening of sleep architecture. This included increased awake time and decreased REM sleep. For the group, the overnight decrement in FEV1 was not improved. Of interest, two patients did have a marked improvement in FEV1 associated with improved oxygen saturation on the CPAP night. These individuals were restudied only on supplemental oxygen. This intervention also improved the overnight FEV1 and allowed the patients to have better sleep compared to the CPAP night. We concluded that CPAP is associated with disrupted sleep architecture in nonapneic asthmatics and nocturnal oxygen desaturation may play a role in the development of nocturnal asthma.

Assessment of bronchodilator response to a beta-adrenergic delivered from an ultrasonic nebulizer.

We assessed the bronchodilator response to a recently available ultrasonic nebulizer (USN) in a population of 17 stable asthmatic patients. These patients were also evaluated for bronchodilator response to two other aerosol delivery systems routinely used in standard clinical practice, the metered-dose inhaler (MDI) and the jet nebulizer (JN). Albuterol was administered from each of the delivery systems in the following manner: MDI, two actuations (180 micrograms); JN, 0.5 ml of 0.5 percent solution (2.5 mg) in 1.0 ml saline solution diluent; and USN, 0.5 ml of 0.5 percent solution (2.5 mg) in 2.5 ml saline solution diluent. Patients demonstrated significant bronchodilator responses to all three delivery systems (p less than 0.0001). The USN produced a greater percentage of increase in FEV1 15 minutes after treatment with albuterol (35.8 +/- 2.3 percent) than either the MDI (18.2 +/- 3.4 percent) or JN (20.8 +/- 3.1 percent) (p less than 0.005). The mean percentage of increase in FE1 observed over a 4-h period after treatment was also greater for the USN (26.5 +/- 2.5 percent) than either the MDI (18.8 +/- 1.8 percent) or JN (15.0 +/- 1.6 percent) (p less than 0.001). We conclude that the USN yields effective bronchodilation in a population of stable asthmatics.

Serum lidocaine concentrations in asthmatics undergoing research bronchoscopy.

STUDY OBJECTIVES: To determine how often serum lidocaine concentrations (SLC) fall into the potentially toxic range (> 5 mg/L) in asthmatics undergoing research bronchoscopy, and to determine whether subject or procedure characteristics are associated with higher SLC. DESIGN: Prospective, observational study. SETTING: Academic research center. PARTICIPANTS: Fifty-one volunteers with mild to moderate asthma enrolled in three separate bronchoscopy protocols to study airway inflammation in asthma. INTERVENTIONS: Lidocaine was administered topically to the upper airway and tracheobronchial tree to achieve local anesthesia for bronchoscopy. Venous blood was sampled during bronchoscopy, 30 min after upper airway anesthesia was completed (time 1), and 30 min after bronchoscopy was completed (time 2). MEASUREMENTS AND RESULTS: The mean total amount of lidocaine administered was 600 +/- 122 mg (8.2 +/- 2.0 mg/kg). No signs or symptoms of lidocaine toxicity were observed in any of the subjects. SLC ranged between 0.10 and 2.90 mg/L at time 1 and 0.50 and 3.20 mg/L at time 2. SLC was significantly correlated with the total amount of lidocaine (milligrams/kilogram) administered at both points (time 1, r = 0.33, p = 0.021; time 2, r = 0.33, p = 0.023). No statistically significant relationship was observed between SLC and subject age, sex, weight, baseline FEV(1), procedure length, or study protocol. No statistically significant relationship was found between subject FEV(1) and either total lidocaine dose or procedure length. CONCLUSIONS: An average total dose of 600 mg (8.2 mg/kg) of lidocaine appears to be safe in mild to moderate asthmatics undergoing research bronchoscopy.

Results of decortication in chronic empyema with special reference of paragonimiasis.

During a recent 6 years period, we managed chronic empyema by decortication, either alone or in combination with thoracoplasty, in 58 patients: In 16 patients, the empyema was a complication of Paragonimus westermani infestation (PWI), in 20, tuberculosis (TB), and in 22, of varied origins. There were no operative deaths, and satisfactory or good results were obtained in 55 patients (94.8%). We believe that the result in a patient in whom thoracoplasty is required should be called satisfactory, not good. Only 55% of the TB group achieved a good result, compared with 89% of the other patients. Of the 10 patients who required a thoracoplasty, seven were in the TB group. Empyema complicating TB is associated with a higher incidence of fistula formation and is more difficult to treat than other empyemas. The empyema associated with PWI develops more slowly, tends to form fewer fistulas, and eventuates in a good result more often than empyema associated with TB. Two of our three poor results stemmed from failure to do a thoracoplasty when the lung did not expand. We recommend thoracoplasty when the lung does not expand well at operation; it should be done at the time of decortication to avoid the necessity for a second major procedure. Chronic empyema is a common complication of paragonimiasis in Korea and by inference in other parts of the world where PWI is endemic. As international travel increases, some patients with PWI and complicating empyema may be seen in the United States. American thoracic surgeons should be aware of this disease, its treatment and its chest complications.

A method for the standardized offline collection of exhaled nitric oxide.

STUDY OBJECTIVES: Exhaled nitric oxide (ENO) is a noninvasive marker of airway inflammation. The purpose of this study was to compare a standardized offline ENO measurement apparatus with a validated on-line method. DESIGN: Asthmatic volunteers (n = 21) had ENO measured by the two following methods: (1) inhalation to total lung capacity (TLC) followed by exhalation at a constant flow (45 mL/s) against a high resistance, while monitoring nitric oxide (NO) and pressure on-line; and (2) inhalation to TLC and exhalation into mylar balloons via an apparatus that included the same resistance and flow rate as used in the on-line method. We also examined NO stability in mylar balloons over 48 h. MEASUREMENTS AND RESULTS: ENO values (given as geometric mean in parts per billion [ppb]; 95% confidence intervals) differed between the on-line method (69.6; 42.6 to 113.8) and the offline method (49.5; 30.9 to 79.3), indicating that the offline method gave lower ENO measures than the on-line method (p < 0.001). Furthermore, this difference between measures increased with increasing mean values. The intraclass correlation coefficient (0.931), however, showed excellent correlation between the on-line and offline methods. Within-subject repeatability, as assessed by the coefficient of repeatability (CR), was good for both the on-line and offline methods (CR, 1.09 and 1.17, respectively). Geometric mean NO concentrations (95% confidence limits) in mylar balloons containing exhalate increased from a baseline of 55.8 ppb (36.9, 84.4) to 64.5 ppb (45.6, 91.1) and 69.5 ppb (51.4, 94.0) at 24 h and 48 h, respectively. CONCLUSIONS: The offline method gave reproducible ENO values that were consistently smaller than, but showed good correlation with, values obtained with on-line ENO collection. This method is suitable for offline collection, but the measured values are not interchangeable with those obtained by on-line measurement.

Exhaled nitric oxide correlated with induced sputum findings in COPD.

STUDY OBJECTIVES: Neutrophilic airway inflammation may underlie the pathogenesis of COPD. We examined repeated measurements of the fractional concentration of exhaled nitric oxide (FENO) and the correlation with cells and mediators in induced sputum (IS) from patients with COPD. PARTICIPANTS: Eleven COPD subjects (9 men and 2 women, aged 46 to 69 years) with predicted FEV(1) of 45 to 70%. SETTING: A hospital research laboratory. DESIGN: Single-cohort, prospective study with four visits at two weekly intervals. INTERVENTIONS: FENO and spirometry were assessed at all visits, and IS for differential cell count, leukotriene-B(4) (LTB(4)) and interleukin (IL)-8, nitrite, and nitrate at visit 1, visit 3, and visit 4. RESULTS: During the study, there were significant declines in mean percent predicted FEV(1), from 55.2 to 51.6% (p = 0.029), and mean FEV(1)/FVC ratio, from 50.4 to 45.4% (p = 0.001), accompanied by a significant increase in FENO geometric mean (95% confidence limits), from 15.2 (10.9 to 21.2) to 23.6 (17.1 to 32.4) parts per billion (p = 0.037), and sputum LTB(4), from 1.79 (1.03 to 3.11) to 3.57 (1.95 to 6.53) ng/mL (p = 0.033), but no significant change in other sputum parameters. From visits 1 to 4, the change in percent neutrophils correlated with the changes in FENO and IL-8 (r = 0.648, p = 0.028; r = 0.60, p = 0.05, respectively). Hypertonic saline solution induction of sputum caused a fall in FEV(1), from 1.83 +/- 0.44 to 1.46 +/- 0.44 L (p = 0.049). CONCLUSIONS: The worsening spirometry results were accompanied by significant increases in FENO and sputum LTB(4). FENO may be related to neutrophilic inflammation driven by the chemoattractant IL-8. FENO and IS may be useful markers of airway inflammation in COPD patients. Sputum induction with hypertonic saline solution causes a significant fall in FEV(1) requiring appropriate caution.

Methotrexate or total lymphoid radiation for treatment of persistent or recurrent allograft cellular rejection: a comparative study.

BACKGROUND: Methotrexate and total lymphoid irradiation (TLI) have been used successfully for treatment of recurrent and persistent rejection in orthotopic heart transplant recipients; however, there has been no comparison of these two modalities. METHODS: We retrospectively compared the efficacy of methotrexate (n = 29) versus TLI (n = 28) in heart transplant recipients with recurrent or persistent rejection. All patients received induction therapy (rabbit anti-thymocyte globulin or OKT3) and standard triple immunosuppressive therapy. Methotrexate (7.5 mg to 22.5 mg per wk) or TLI (80 cGy x 10 fractions) was used for the treatment of recurrent or persistent rejection on the basis of clinical indications. Average biopsy scores (International Society of Heart and Lung Transplantation biopsy score/total number of biopsies performed) calculated over 3-month periods, daily maintenance prednisone dose before and after methotrexate or TLI treatment, and actuarial survival and freedom from angiographic coronary artery disease and infection were compared. To control for the general decrease in prednisone with increased time from transplantation, a control group matched for time from transplantation was selected. RESULTS: Recipient sex and age at transplant, donor age, and donor ischemic time were similar in both groups. Days after transplantation to start of therapy was longer in patients receiving methotrexate; however, this did not reach statistical significance. Patients receiving TLI had received more cumulative corticosteroids and OKT3 before the start of TLI therapy (p < 0.001). There were no differences in actuarial freedom from infection or coronary artery disease between the two groups and between the treatment groups and the control group. Actuarial survival was reduced in patients receiving TLI 3 years after transplantation (p < 0.05). Maintenance prednisone doses from 3 months before until 9 months after therapy (mg/kg) were not different between patients receiving TLI and methotrexate and were significantly greater than the prednisone doses in the control group. Four months after treatment initiation, the prednisone dose was significantly reduced in both treatment groups compared with the pretherapy dose (methotrexate 0.28 +/- 0.16 to 0.22 +/- 0.13, p = 0.05; TLI 0.36 +/- 0.16 to 0.22 +/- .07, p < 0.001). The average biopsy score was significantly reduced by both methotrexate and TLI therapy (methotrexate 1.8 +/- 0.7 to 0.83 +/- 0.9, p = 0.0001; TLI 2.1 +/- 0.8 to 1.0 +/- 0.9, p = 0.0001). CONCLUSION: Methotrexate and TLI are both effective for the treatment of recurrent or persistent rejection after heart transplantation, reducing average biopsy scores and daily maintenance prednisone doses. There was a reduction in actuarial survival rates in patients treated with TLI, possibly reflecting the greater rejection therapy received before TLI initiation. Because both agents are effective, the choice of methotrexate or TLI may be based on clinical indications, as well as other issues, such as cost, compliance, and availability.