Revision of total shoulder arthroplasty to hemiarthroplasty: results at mean 5-year follow-up.

BACKGROUND: Although there is a trend to manage failed anatomic total shoulder arthroplasties (aTSA) with revision to a reverse total shoulder arthroplasty, such revisions can be complicated by difficulties in baseplate fixation, instability, and acromial stress fractures. Some cases of failed aTSA may be safely revised to a hemiarthroplasty (HA). The objectives of this study were to report patient-reported outcomes after conversion from aTSA to HA and assess patient and shoulder characteristics associated with a successful outcome. METHODS: Patients who underwent revision from aTSA to HA between 2009 and 2018 were contacted. Patient demographics, surgical history, intraoperative findings, and microbiology results were collected. Patient-reported outcomes were collected with minimum 2-year follow-up. Preoperative radiographic characteristics were reviewed for component positioning and component loosening. Patients with a clinically significant improvement exceeding the minimal clinically important difference (MCID) of the Simple Shoulder Test (SST) were compared with those patients who did not improve past the MCID. RESULTS: Twenty-nine patients underwent conversion from aTSA to HA with a mean follow-up of 4.5 ± 1.8 years. Intraoperative glenoid or humeral component loosening was found in all 29 patients. Pain improved in 25 of 30 patients (87%), and mean pain scores improved from 6.2 ± 2.3 to 3.1 ± 2.4 (P < .001). SST scores improved from 4.1 ± 3.1 to 7.3 ± 3.2 (P < .001), and 18 of 29 patients (62%) had improvement above the SST MCID threshold of 2.4. The mean American Shoulder and Elbow Surgeons score at the latest follow-up was 64 ± 19, and the Single Assessment Numeric Evaluation score was 65 ± 23. Twenty-two of 29 (76%) patients were satisfied with the procedure. Four patients (14%) required conversion to total shoulder arthroplasty-2 to anatomic and 2 to reverse. An additional 3 patients (10%) had a revision HA performed. No significant differences in patient or shoulder characteristics were found in those patients who improved greater than the MCID of the SST compared patients who improved less than the MCID of the SST. Fifty-nine percent of patients had ≥2 positive cultures with the same bacteria, and 82% of these were with Cutibacterium. Seven of 8 patients (88%) with a loose humeral component had ≥2 positive cultures with the same bacteria. DISCUSSION: Component loosening is a common failure mode after aTSA. Revision to HA can improve pain and patient-reported outcomes in most patients.

The minimal clinically important differences of the Simple Shoulder Test are different for different arthroplasty types.

BACKGROUND: Preoperative and postoperative patient self-reported measures are the key to understanding the benefit of shoulder arthroplasty for patients with different diagnoses and having different surgical approaches. The minimal clinically important difference (MCID) for patient-reported outcomes such as the Simple Shoulder Test (SST) is often used to document the amount of improvement that is of importance to the patient; however, the MCID may differ for different types of shoulder arthroplasty. The objective of this study was to report the MCID of the SST and the MCID of the percentage of maximal possible improvement (%MPI) for 5 different arthroplasty types. METHODS: Eight hundred eighty-seven patients undergoing shoulder arthroplasty with preoperative SST scores, 2-year postoperative SST scores, and patient satisfaction were included. The sample comprised 368 patients undergoing anatomic total shoulder arthroplasty (aTSA), 330 patients undergoing ream-and-run arthroplasty (R&R), 80 patients undergoing reverse total shoulder arthroplasty (rTSA), 53 patients undergoing cuff tear arthropathy arthroplasty, and 56 patients undergoing hemiarthroplasty. For each type of arthroplasty, the anchor-based method was used for calculating the MCID for both absolute SST scores and %MPI. RESULTS: Significant improvements in SST values were seen for all arthroplasty types. The MCID for SST change was 2.3 overall but ranged from 1.6 for aTSA, to 2.6 for R&R, to 3.7 for rTSA. The MCID for %MPI was 32% overall but ranged from 22% for aTSA to 42% for hemiarthroplasty. The percentage of patients exceeding the MCID threshold was highest for aTSA at 96% and lowest for hemiarthroplasty at 61%. CONCLUSION: The same MCID value may not be appropriate for different types of shoulder arthroplasty. This study reports MCID thresholds that can be used when assessing the effectiveness for each of the common types of shoulder arthroplasty.

Combined orthodontic and surgical open bite correction.

OBJECTIVES: To examine the stability of combined surgical and orthodontic bite correction with emphasis on open-bite closure. All study patients were treated with strict and consistent orthodontic and surgical protocols. MATERIALS AND METHODS: Study inclusion required all patients to have anterior open bites, maxillary accentuated curve of Spee, 36-month minimum follow-up, and no temporomandibular joint pathology. Thirty patients met the inclusion/exclusion criteria. Importantly, segmental upper arch orthodontic preparation (performed by EG) was used. Surgery consisted of a multisegment Le Fort I (MSLFI) combined with a bilateral sagittal osteotomies (BSSO). Surgery was performed (by ADA and LT) at the Department of Dentistry and Maxillofacial Surgery of the University of Verona, Italy. RESULTS: The long-term open bite and overjet relapse were not statistically significant. The mean transverse relapse of the upper and lower molars was statistically significant. Of great importance, the upper and lower arch widths narrowed together, maintaining intercuspation of the posterior dentition which prevented anterior open bites from developing. CONCLUSIONS: This study revealed stability of three-dimensional occlusal correction including anterior open bite. Stable open bite closure was achieved by using rigid protocols for orthodontic preparation, surgical techniques, surgical follow-up, and orthodontic finishing.

Reverse shoulder arthroplasty yields similar results to anatomic total shoulder arthroplasty for the treatment of humeral head avascular necrosis.

BACKGROUND: Avascular necrosis (AVN) of the humeral head frequently results in humeral head collapse and end-stage arthritic changes of the glenohumeral joint. Despite the recent proliferation of reverse total shoulder arthroplasty (RTSA), reports on the use of RTSA for AVN remain limited. The purpose of this study was to document the outcomes of shoulders indicated for RTSA in the setting of humeral head AVN and compare these with AVN shoulders indicated for the gold standard, anatomic total shoulder arthroplasty (aTSA). METHODS: A retrospective review of a multinational shoulder arthroplasty database was performed between August 2005 and August 2017. All shoulders with a preoperative diagnosis of AVN (aTSA in 52 and RTSA in 67) were reviewed. The shoulders in the RTSA cohort were matched (1:1) to shoulders with cuff tear arthropathy, whereas the shoulders in the aTSA cohort were matched (1:1) to shoulders with primary osteoarthritis. The mean follow-up period was 47 months (range, 24-130 months) for RTSA and 54 months (range, 24-124 months) for aTSA. Shoulders were evaluated for active range of motion (ROM) and patient-reported outcome measures (PROMs) prior to surgery and at latest follow-up. Patients treated with RTSA were compared with both the aTSA study cohort and the control group using the Student t test or χ2 test as indicated. RESULTS: RTSAs performed for AVN demonstrated significant improvements in all ROMs and PROMs. Patients undergoing aTSA for AVN were significantly younger than those undergoing RTSA (59 years vs. 73 years, P < .001). At similar follow-up points, the RTSA cohort demonstrated significantly greater improvement in abduction (+51° vs. +32°, P = .03) whereas the aTSA cohort demonstrated significantly greater improvement in internal rotation. Postoperative University of California, Los Angeles scores (30 vs. 27, P = .014) and visual analog scale scores (1.4 vs. 2.4, P = .025) were better after RTSA; however, these differences between prosthesis types did not exceed the minimal clinically important difference. When compared with the control patients, the patients undergoing RTSA for AVN showed similar improvements in all ROMs and PROMs. Similarly, aTSA performed for AVN resulted in comparable improvements in pain, ROMs, and PROMs compared with aTSA performed for primary osteoarthritis. CONCLUSION: RTSA results in similar PROMs to aTSA in the treatment of AVN. Therefore, surgeons should continue to consider other patient factors such as glenoid bone loss and rotator cuff status when selecting implant polarity in patients with AVN.

Retrocopying expands the functional repertoire of APOBEC3 antiviral proteins in primates.

Host-virus arms races are inherently asymmetric; viruses evolve much more rapidly than host genomes. Thus, there is high interest in discovering mechanisms by which host genomes keep pace with rapidly evolving viruses. One family of restriction factors, the APOBEC3 (A3) cytidine deaminases, has undergone positive selection and expansion via segmental gene duplication and recombination. Here, we show that new copies of A3 genes have also been created in primates by reverse transcriptase-encoding elements like LINE-1 or endogenous retroviruses via a process termed retrocopying. First, we discovered that all simian primate genomes retain the remnants of an ancient A3 retrocopy: A3I. Furthermore, we found that some New World monkeys encode up to ten additional APOBEC3G (A3G) retrocopies. Some of these A3G retrocopies are transcribed in a variety of tissues and able to restrict retroviruses. Our findings suggest that host genomes co-opt retroelement activity in the germline to create new host restriction factors as another means to keep pace with the rapid evolution of viruses. (163).

Combinatorial mutagenesis of rapidly evolving residues yields super-restrictor antiviral proteins.

Antagonistic interactions drive host-virus evolutionary arms races, which often manifest as recurrent amino acid changes (i.e., positive selection) at their protein-protein interaction interfaces. Here, we investigated whether combinatorial mutagenesis of positions under positive selection in a host antiviral protein could enhance its restrictive properties. We tested approximately 700 variants of human MxA, generated by combinatorial mutagenesis, for their ability to restrict Thogotovirus (THOV). We identified MxA super-restrictors with increased binding to the THOV nucleoprotein (NP) target protein and 10-fold higher anti-THOV restriction relative to wild-type human MxA, the most potent naturally occurring anti-THOV restrictor identified. Our findings reveal a means to elicit super-restrictor antiviral proteins by leveraging signatures of positive selection. Although some MxA super-restrictors of THOV were impaired in their restriction of H5N1 influenza A virus (IAV), other super-restrictor variants increased THOV restriction without impairment of IAV restriction. Thus, broadly acting antiviral proteins such as MxA mitigate breadth-versus-specificity trade-offs that could otherwise constrain their adaptive landscape.

What Is the Survivorship of Revision Surgery Performed for the Chronically Dislocated THA?

BACKGROUND: Instability remains one of the most common indications for revision THA. However, little is known about the efficacy of surgery for and the complications associated with revision THA for patients with a chronically dislocated THA, which we define as a dislocation of more than 4 weeks. QUESTIONS/PURPOSES: For patients with a chronically dislocated THA undergoing revision THA, we asked (1) What is the survivorship free from additional revision for these procedures? (2) What complications are associated with revision THA in this setting? (3) What are the clinical outcomes as measured by the Harris hip score in these procedures? METHODS: From 1998 to 2014, 1084 patients who underwent revision THA for instability were reviewed and 33 patients (33 hips) were identified who had a hip that had been dislocated for more than 4 weeks. Median time dislocated was 4 months (range, 1-120 months), and the mean distance of the femoral head above hip center at presentation was 45 mm. Mean patient age was 67 ± 17 years, and 79% of patients (26 of 33) were women. During the period in question, we used four approaches: Treatment with acetabular component revision in 18 of 33 patients (55%), head and liner exchange in nine patients (27%), both-component revision in five patients (15%), and isolated femoral component revision in one patient (3%). A constrained liner was used in 17 patients (52%), including six of the patients treated with acetabular component revision, and three of those who had both-component revisions. During the period in question, our general indications were hip pain and/or unacceptable function with the chronically dislocated prosthesis. Our sample size was too small to evaluate the association of the procedure choice on survivorship or complication risk. We used Kaplan-Meier survivorship analysis to estimate survivorship free from complication, reoperation, or revision. Mean followup was 4.4 years (range, 2-10 years). RESULTS: Survivorship free from any revision, complication, or reoperation was 61% at 5 years (95% CI, 43-82). Survivorship free from revision was 83% at 5 years (95% CI, 67-98). Etiology for revision was aseptic loosening in three of 33 hips (9%), recurrent dislocation in two hips (6%), and deep periprosthetic joint infection in two hips (6%). Five complications (15%) did not result in a reoperation, including one dislocation and one incomplete peroneal nerve palsy in a patient after an anterolateral approach. The Harris hip score improved from mean 50 ± 17 preoperatively to mean 80 ± 11 at 5 years. CONCLUSIONS: Chronically dislocated THAs can be successfully managed with revision THA. We recommend close evaluation of the components for aseptic loosening, performing revision surgery only on patients with pain and poor function, and thoroughly counseling patients that survivorship is modest and complications are common. LEVEL OF EVIDENCE: Level IV, therapeutic study.

Intraobserver and interobserver reliability of the modified Walch classification using radiographs and computed tomography.

BACKGROUND: The Walch classification was introduced to classify glenoid morphology in primary glenohumeral osteoarthritis. A modified Walch classification was recently proposed, with 2 additional categories, B3 (monoconcave glenoid with posterior bone loss leading to retroversion > 15° or subluxation > 70%) and D (excessive anterior subluxation), as well as a more precise definition of subtypes A2 and C. The purpose of this study was to evaluate the intraobserver and interobserver agreement of the modified Walch classification system using both plain radiographs and computed tomography (CT). METHODS: Three fellowship-trained shoulder surgeons blindly and independently evaluated radiographs and CT scans of 100 consecutive shoulders (98 patients) with primary glenohumeral osteoarthritis and classified all shoulders according to the modified Walch classification in 4 separate sessions, each 4 weeks apart. Statistical analysis with the κ coefficient was used to evaluate reliability. RESULTS: The first reading by the most senior observer on the basis of CT scans was used as the gold standard (distribution: A1, 18; A2, 12; B1, 20; B2, 25; B3, 22; C, 1; and D, 2). The average intraobserver agreement for radiographs and CT scans was 0.73 (substantial; 0.72, 0.74, and 0.72) and 0.73 (substantial; 0.77, 0.69, and 0.72), respectively. The average interobserver agreement was 0.55 (moderate; 0.61, 0.51, and 0.53) for radiographs and 0.52 (moderate; 0.63, 0.50, and 0.43) for CT scans. CONCLUSION: Intraobserver agreement of the modified Walch classification was substantial both for axillary radiographs and for CT scans. Interobserver agreement was fair. Although the modified Walch classification represents an improvement over the original classification, automated computer-based analysis of CT scans may be needed to further improve the value of this classification.

Long-Term Outcomes of Glenohumeral Arthrodesis.

BACKGROUND: Glenohumeral arthrodesis is associated with a high rate of complications. Although patients experience reasonable pain relief and shoulder stability, they experience marked limitations in their upper-extremity function. The purpose of this study was to examine the clinical outcomes of glenohumeral arthrodesis. METHODS: Twenty-nine patients with 29 affected shoulders underwent primary glenohumeral arthrodesis between 1992 and 2009. Surgical indications included rotator cuff arthropathy and pseudoparalysis (n = 7), neurologic injuries (n = 12), chronic infection (n = 3), recurrent dislocations (n = 3), and proximal humeral or shoulder girdle tumors (n = 4). Surgical fixation techniques included plates and screws in 18 patients and screws only in 11 patients. RESULTS: All patients were examined, with a mean follow-up of 12 years (range, 2 to 22 years). Twelve patients (41%) had postoperative complications, including 6 periprosthetic fractures, 7 nonunions, and 3 infections. Eleven patients (38%) required additional surgical procedures after arthrodesis, including revision internal fixation to achieve glenohumeral fusion after nonunions (n = 7), irrigation and debridement with antibiotic treatment for deep infections (n = 2), open reduction and internal fixation to treat fracture (n = 2), and implant removal to treat symptomatic patients (n = 3). Patients experienced reasonable overall pain relief. The mean postoperative scores were 35 points for the Subjective Shoulder Value, 58 points for the Disabilities of the Arm, Shoulder and Hand (DASH) questionnaire, and 54 points for the Short Form-36. Eighty-seven percent of patients reported postoperative limitations. Patients with neurologic injuries had worse functional outcomes, and an arthrodesis position of ≥25° yielded better functional outcomes. CONCLUSIONS: Glenohumeral arthrodesis is associated with a high rate of patients with complications (41%). Although patients experience reasonable pain relief and shoulder stability, they experience marked limitations in their upper-extremity function. LEVEL OF EVIDENCE: Therapeutic Level IV. See Instructions for Authors for a complete description of levels of evidence.

Bony Versus Soft Tissue Reconstruction for Anterior Shoulder Instability: An Expected Value Decision Analysis.

BACKGROUND: One complication of anteroinferior glenohumeral shoulder dislocation is a critical bone defect that requires surgical repair to prevent recurrent instability. However, controversy exists regarding the surgical management because both open and arthroscopic surgeries have respective advantages and disadvantages. Moreover, it is difficult to determine the patient's preferred treatment, as factors that influence treatment choice include recurrence rates, morbidity of the procedures, and patient preferences. HYPOTHESIS: Patients who have a higher probability of recurrent instability after arthroscopic surgery will select open surgery whereas patients with a lower probability of recurrent instability after arthroscopic surgery will favor arthroscopy. STUDY DESIGN: Economic and decision analysis; Level of evidence, 2. METHODS: A decision tree was constructed to model each hypothetical outcome after open or arthroscopic surgery for glenohumeral instability in patients with bone defects. A literature review was performed to determine the probability of occurrence for each node while utility values for each outcome were obtained via patient-administered surveys given to 50 patients without prior history of shoulder injury or dislocation. Fold-back analysis was then performed to show the optimal treatment strategy. Finally, sensitivity analysis established the thresholds at which open treatment becomes the optimal treatment. RESULTS: The ultimate expected value-the objective evaluation of all potential outcomes after choosing either open or arthroscopic surgery-was found to be greater for arthroscopic surgery than for open surgery (87.17 vs 81.64), indicating it to be the preferred treatment. Results of sensitivity analysis indicated that open surgery becomes the preferred treatment when probability of recurrence after arthroscopic treatment is ≥23.8%, although varying the utility, defined as an aggregate patient preference for a particular outcome, has no effect on the model. When the rate of no complication after open surgery is 97.6%, open surgery becomes the patient's preferred treatment. CONCLUSION: Arthroscopic surgery is an acceptable treatment if recurrent instability occurs consistently at ≤23.8%. This has important implications given the technical difficulty of successfully performing arthroscopic fixation to resolve recurrent anteroinferior glenohumeral dislocations associated with critical osseous defects. However, due to a lack of clinical outcomes studies, more research is needed to better predict the optimal operative treatment.

Right ventricular gunshot wound with retrograde embolization.

There have been numerous reports concerning gunshot wounds to the heart over the years. Many reports discuss bullets that have embolized and have migrated antegrade. However, there has never been a case reported on the retrograde embolization of a bullet from the right ventricle into the inferior vena cava. This case report involves a 15-year-old boy who was accidentally shot in the chest. The bullet entered at the mid-manubrial area, and penetrated the anterior wall of the right ventricle causing a tamponade. A chest x-ray film confirmed a bullet in the right ventricle. The patient was stabilized in the emergency department, and taken to the operating room for an emergent mediastinal exploration with evacuation of pericardial tamponade and repair of the right ventricle. After the tamponade was relieved, a Trans-Esophageal Echocardiogram was performed to locate the bullet, which could not be found in the ventricle. Chest and abdominal radiography were performed to locate the bullet. X-ray films confirmed that the bullet had migrated retrograde down into the inferior vena cava. Interventional radiology and vascular surgery departments were consulted. The consensus was to snag the bullet under fluoroscopic guidance, and pull it down into the right femoral vein for easy retrieval. The chest was closed and the patient was transferred from the surgery department to the interventional radiology department. Under fluoroscopy, the bullet was pulled down into the right common femoral vein. The bullet was extracted per venorrhaphy. The patient was extubated within hours after surgery and discharged home within 72 hours of surgery.

The effects of sodium sulfate, glycosaminoglycans, and Congo red on the structure, stability, and amyloid formation of an immunoglobulin light-chain protein.

Light-chain amyloidosis (AL) is characterized by immunoglobulin light-chain fragments aggregating into amyloid fibrils that deposit extracellularly in vital organs such as the kidney, the heart, and the liver, resulting in tissue degeneration and organ failure, leading to death. Cardiac involvement is found in 50% of AL patients and presents the most severe cases with a life expectancy of less than a year after diagnosis. In this study, we have characterized the variable domain of a cardiac AL patient light chain called AL-09. AL-09 folds as a beta-sheet and is capable of forming amyloid fibrils both in the presence of sodium sulfate and in self-seeded reactions under physiological conditions. Glycosaminoglycans such as dermatan sulfate and heparin promote amyloid formation of self-seeded AL-09 reactions, while the glycosaminoglycan chondroitin sulfate A stabilized oligomeric intermediates and did not elongate the preformed fibrils (nucleus) present in the reaction. Finally, the histological dye Congo red, known to bind to the cross beta-sheet structure of amyloid fibrils, inhibits AL-09 amyloid fibril formation in the presence of sodium sulfate and in self-seeded reactions. This paper provides insight into the impact of different reagents on light-chain stability, structure, amyloid fibril formation, and inhibition.

The cAMP-specific phosphodiesterase 4B mediates Abeta-induced microglial activation.

Microglial activation is a key player in the degenerative process that accompanies the deposition of amyloid-beta (Abeta) peptide into senile plaques in Alzheimer's disease (AD) patients. The goal of this study is to identify novel genes involved in microglial activation in response to Abeta peptide. Prompted by the fact that soluble Abeta(1-42) (sAbeta(1-42))-stimulated primary rat microglia produce more tumor necrosis factor-alpha (TNF-alpha) than fibrillar Abeta(1-42) (fAbeta(1-42))-stimulated microglia, we examined gene expression in these cells following stimulation using cDNA arrays. This analysis confirms the upregulation caused by both sAbeta(1-42) and fAbeta(1-42) of pro-inflammatory molecules such as TNF-alpha, interleukin-1beta and macrophage inflammatory protein-1alpha. In addition, other transcripts not previously described in the context of Abeta-induced microglial activation were identified. The modulation of some of these genes within microglial cells seems to be specific to sAbeta(1-42) as compared to fAbeta(1-42) suggesting that different forms of Abeta may activate distinct pathways during the progression of AD. Importantly, we demonstrate that Pde4B, a cAMP-specific phosphodiesterase, is upregulated by Abeta and results in an increased production of TNF-alpha. Inhibition of Pde4B reduces by up to 70% the release of TNF-alpha from sAbeta-stimulated microglial cells, implicating cAMP as an important mediator of Abeta-induced microglial activation.

Human thiopurine S-methyltransferase pharmacogenetics: variant allozyme misfolding and aggresome formation.

Thiopurine S-methyltransferase (TPMT) catalyzes the S-methylation of thiopurine drugs. TPMT genetic polymorphisms represent a striking example of the potential clinical value of pharmacogenetics. Subjects homozygous for TPMT*3A, the most common variant allele for low activity, an allele that encodes a protein with two changes in amino acid sequence, are at greatly increased risk for life-threatening toxicity when treated with standard doses of thiopurines. These subjects have virtually undetectable levels of TPMT protein. In this study, we tested the hypothesis that TPMT*3A might result in protein misfolding and aggregation. We observed that TPMT*3A forms aggresomes in cultured cells and that it aggregates in vitro, functional mechanisms not previously described in pharmacogenetics. Furthermore, there was a correlation among TPMT half-life values in rabbit reticulocyte lysate, aggresome formation in COS-1 cells, and protein aggregation in vitro for the three variant allozymes encoded by alleles that include the two TPMT*3A single-nucleotide polymorphisms. These observations were compatible with a common structural explanation for all of these effects, a conclusion supported by size-exclusion chromatography and CD spectroscopy. The results of these experiments provide insight into a unique pharmacogenetic mechanism by which common polymorphisms affect TPMT protein function and, as a result, therapeutic response to thiopurine drugs.

Colloidal gold: a novel nanoparticle vector for tumor directed drug delivery.

Colloidal gold, a sol comprised of nanoparticles of Au(0), has been used as a therapeutic for the treatment of cancer as well as an indicator for immunodiagnostics. However, the use of these gold nanoparticles for in vivo drug delivery has never been described. This communication outlines the development of a colloidal gold (cAu) nanoparticle vector that targets the delivery of tumor necrosis factor (TNF) to a solid tumor growing in mice. The optimal vector, designated PT-cAu-TNF, consists of molecules of thiol-derivatized PEG (PT) and recombinant human TNF that are directly bound onto the surface of the gold nanoparticles. Following intravenous administration, PT-cAu-TNF rapidly accumulates in MC-38 colon carcinoma tumors and shows little to no accumulation in the livers, spleens (i.e., the RES) or other healthy organs of the animals. The tumor accumulation was evidenced by a marked change in the color of the tumor as it acquired the bright red/purple color of the colloidal gold sol and was coincident with the active and tumor-specific sequestration of TNF. Finally, PT-cAu-TNF was less toxic and more effective in reducing tumor burden than native TNF since maximal antitumor responses were achieved at lower doses of drug.

Enhancement of glutathione cardioprotection by ascorbic acid in myocardial reperfusion injury.

The present experiment determined the effects of glutathione and ascorbic acid, the two most important hydrophilic antioxidants, on myocardial ischemia-reperfusion injury and evaluated their relative therapeutic values. Isolated rat hearts were subjected to ischemia (30 min) and reperfusion (120 min) and treated with ascorbic acid, glutathione monoethyl ester (GSHme), or their combination at the onset of reperfusion. Administration of 1 mM GSHme alone, but not 1 mM ascorbic acid alone, significantly attenuated postischemic injury (P < 0.05 versus vehicle). Most interestingly, coadministration of ascorbic acid with GSHme markedly enhanced the protective effects of GSHme (P < 0.01 versus vehicle). The protection exerted by the combination of GSHme and ascorbic acid at 1 mM each was significantly greater than that observed with 1 mM GSHme alone (P < 0.05). Moreover, treatment with GSHme alone or GSHme plus ascorbic acid markedly reduced myocardial nitrotyrosine levels, suggesting that these treatments attenuated myocardial peroxynitrite formation. These results demonstrated that 1) GSHme, but not ascorbic acid, exerted protective effects against ischemia-reperfusion injury; and 2) the protective effects of GSHme were further enhanced by coadministration with ascorbic acid, suggesting a synergistic effect between GSHme and ascorbic acid.

Selective cytotoxicity of intracellular amyloid beta peptide1-42 through p53 and Bax in cultured primary human neurons.

Extracellular amyloid beta peptides (Abetas) have long been thought to be a primary cause of Alzheimer's disease (AD). Now, detection of intracellular neuronal Abeta1--42 accumulation before extracellular Abeta deposits questions the relevance of intracellular peptides in AD. In the present study, we directly address whether intracellular Abeta is toxic to human neurons. Microinjections of Abeta1--42 peptide or a cDNA-expressing cytosolic Abeta1--42 rapidly induces cell death of primary human neurons. In contrast, Abeta1--40, Abeta40--1, or Abeta42--1 peptides, and cDNAs expressing cytosolic Abeta1--40 or secreted Abeta1--42 and Abeta1--40, are not toxic. As little as a 1-pM concentration or 1500 molecules/cell of Abeta1--42 peptides is neurotoxic. The nonfibrillized and fibrillized Abeta1--42 peptides are equally toxic. In contrast, Abeta1--42 peptides are not toxic to human primary astrocytes, neuronal, and nonneuronal cell lines. Inhibition of de novo protein synthesis protects against Abeta1--42 toxicity, indicating that programmed cell death is involved. Bcl-2, Bax-neutralizing antibodies, cDNA expression of a p53R273H dominant negative mutant, and caspase inhibitors prevent Abeta1--42-mediated human neuronal cell death. Taken together, our data directly demonstrate that intracellular Abeta1--42 is selectively cytotoxic to human neurons through the p53--Bax cell death pathway.