Laterality of lumbar disc herniation.

BACKGROUND: Lumbar disc herniation (LDH) is most common in men aged 20-40 at the L4/5 level; however, left-right differences have not been reported. Improving our understanding of left- and right-side LDH may facilitate the estimation of mechanical load on intervertebral discs. Here, we assessed left-right differences in LDH via a retrospective analysis of LDH cases. METHODS: Among 10,972 surgical cases of LDH identified in the Nagoya Spine Group database, 2899 in which right- and left-LDH sides were observable in a single vertebral segment were identified (mean age 46.3 ± 16.6 years, 2028 males). The following characteristics of patients with right- and left-LDH were compared: age, LDH level, surgical technique, operative time, blood loss, length of hospital stay, preoperative Japan Orthopaedic Association (JOA) score, and JOA recovery rate. RESULTS: LDH occurred on the right and left sides in 1358 and 1541 patients, respectively, with patients with right-side LDH significantly older than those with left (47.9 ± 16.6 versus 45.0 ± 16.5, respectively; p < 0.001). No between-group differences in sex, age, LDH level, surgical technique, operative time, blood loss, length of hospital stay, preoperative JOA score, or JOA recovery rate were observed. The occurrence of right-side LDH increased with age, occurring in 42.7%, 45.1%, 49.9%, and 54.7% of patients aged 10-29, 30-49, 50-69, and 70-89 years, respectively. CONCLUSION: Left-side LDH was observed more frequently than right; however, right-side LDH incidence increased with age. No significant between-group differences regarding symptoms, treatments, or outcomes were observed.

Risk factors of non-union in Anderson-D'Alonzo type III odontoid fractures with conservative treatment.

OBJECTIVE: The purpose of this study was retrospectively to analyze the risk factors for non-union in Anderson-D'Alonzo type III odontoid fractures with conservative treatment. METHODS: 25 patients with type III fractures were analyzed. Coronal and sagittal tilt as well as sagittal and lateral mass gaps were measured by using computed tomography. RESULTS: The non-union group had significantly higher age, greater coronal tilt and lateral mass gap. Especially, the lateral mass gap was >2 mm in all cases with non-union. CONCLUSIONS: Higher age, coronal tilt, and lateral mass gap were significant risk factors for non-union.

Trends in Reoperation for Surgical Site Infection After Spinal Surgery With Instrumentation in a Multicenter Study.

STUDY DESIGN: A multicenter retrospective analysis of a prospectively maintained database. OBJECTIVE: To examine the characteristics of reoperation for surgical site infection (SSI) after spinal instrumentation surgery, including the efficacy of treatment for SSI and instrumentation retention. SUMMARY OF BACKGROUND DATA: Aging of the population and advances in surgical techniques have increased the demand for spinal surgery in elderly patients. Treatment of SSI after this surgery has the main goals of eliminating infection and retaining instrumentation. METHODS: The subjects were 16,707 patients who underwent spine surgery with instrumentation in 11 hospitals affiliated with the Nagoya Spine Group from 2004 to 2015. Details of those requiring reoperations for SSI were obtained from surgical records at each hospital. RESULTS: There were significant increases in the mean age at the time of surgery (54.6-63.7 years) and the number of instrumentation surgeries (726-1977) from 2004 to 2015. The incidence of reoperation for SSI varied from 0.9% to 1.8%, with a decreasing trend over time. Reoperation for SSI was performed in 206 cases (115 men, 91 women; mean age 63.2 years). The average number of reoperations (1.4 vs. 2.3, P < 0.05), time from SSI to first reoperation (4.3 vs. 9.5 days, P < 0.05), and the methicillin-resistant Staphylococcus identification rate (20% vs. 37%, P < 0.01) were all significantly lower in cases with instrumentation retention (n = 145) compared to those with instrumentation removal (n = 61). CONCLUSION: There were marked trends of aging of patients and an increase in operations over the study period; however, the incidences of reoperation and instrumentation removal due to SSI significantly decreased over the same period. Rapid debridement after SSI diagnosis may have contributed to instrumentation retention. These results can serve as a guide for developing strategies for SSI treatment and for improved planning of spine surgery in an aging society. LEVEL OF EVIDENCE: 3.

Spontaneously Resolved Recurrent Cervical Epidural Hematoma in a 37-Week Primigravida.

Study Design Case report. Objective To describe a patient with a recurrent spontaneous spinal epidural hematoma (SSEH) during pregnancy that had spontaneous remission. Methods A 27-year-old primigravida at 37 weeks' gestation suddenly felt a strong left shoulder pain without any trauma. She had a history of fenestration for a spontaneous cervical hematoma when she was 18 years old. An emergency magnetic resonance imaging revealed a recurrence of the cervical epidural hematoma at the C4-T1 level, but she had no paralysis. Results The patient subsequently underwent a cesarean section and delivered a healthy male infant. Her spinal epidural hematoma disappeared. Multislice computed tomography showed no evidence for a vascular malformation or tumor. Three years after the initial cesarean section, she underwent a second one and delivered another male infant. Conclusions We report on a rare case of recurrent SSEH during pregnancy with no neurologic deficits that was treated nonoperatively with close observation and resulted in spontaneous resolution. In such patients with no neurologic deficits, nonoperative management with close observation may be a reasonable alternative.

Comparative study of 2 surgical procedures for osteoporotic delayed vertebral collapse: anterior and posterior combined surgery versus posterior spinal fusion with vertebroplasty.

STUDY DESIGN: Retrospective comparative study. OBJECTIVE: To compare the surgical results of anterior and posterior combined surgery (AP) and posterior fixation with vertebroplasty (VP) for treating osteoporotic delayed vertebral collapse. SUMMARY OF BACKGROUND DATA: The optimal treatment of osteoporotic delayed vertebral collapse has been controversial. Because of aged patients' numerous comorbid medical complications and frequent instrumentation failure secondary to osteoporosis, it is challenging for surgeons to manage osteoporotic delayed vertebral collapse. In spite of this, there have been few reports comparing the surgical results. METHODS: A total of 93 patients with osteoporotic delayed vertebral fracture who underwent spinal surgery were enrolled at 6 hospitals. Sixty-five patients underwent AP surgery in 3 hospitals, and 28 patients underwent VP surgery in the other 3 hospitals. We restricted the spinal-fracture level to thoracolumbar lesion (T10-L2) and excluded patients followed up more than 2 years after surgery. The final numbers of patients included in this study were 24 in the AP group and 21 in the VP group. There were no significant differences between the 2 groups in terms of age, sex, disease duration, or duration of follow-up. RESULTS: Operative time was significantly longer and intraoperative blood loss significantly greater in the AP group. No significant difference between the 2 groups was observed in neurological improvement or the angle of kyphosis correction. However, the loss of correction was significantly greater in the VP group. There were no significant differences in perioperative respiratory or other complications. Implant-related complications and pseudarthrosis were more often observed in the VP group. One patient in the VP group underwent additional surgery for progression kyphosis. CONCLUSION: AP surgery provides stable spinal fixation and reduces implant failure particularly at the thoracolumbar junction because of load bearing of anterior spinal elements. Surgery-related complications in AP surgery were as few in number as with the VP group, and AP surgery is useful for osteoporotic delayed vertebral fracture.

Surgically treated cases of lumbar spondylolysis and isthmic spondylolisthesis: a multicenter study.

STUDY DESIGN: Prospective database study. OBJECTIVES: To grasp the characteristics of surgically treated cases with lumbar spondylolysis or isthmic spondylolisthesis. SUMMARY OF BACKGROUND DATA: A detailed analysis of surgically treated cases with spondylolysis or isthmic spondylolisthesis has never been reported. An epidemiological study in Japan conducted on 2000 subjects found the incidence of lumbar spondylolysis in the Japanese general population (population-based study) to be 5.9% (males: 7.9%, females: 3.9%). Among 124 vertebrae with spondylolysis, there were 0.8% L2 lesions, 3.2% L3 lesions, 5.6% L4 lesions, and 90.3% L5 lesions, including 5 cases (4.3%) with multiple-level lesions. METHODS: We have been registering surgically treated spine cases in our database since 2000. From this database, we prospectively collected cases with lumbar spondylolysis or isthmic spondylolisthesis that were treated surgically between January 2000 and December 2009. We determined the age at surgery, sex, and vertebral level of spondylolysis. RESULTS: Of the 564 spondylolysis patients treated surgically, 66.8% were male and 33.2% were female. The mean age at surgery was 52.5 years (range, 13-84 y). There were 585 vertebrae with spondylolysis including 21 cases (3.7%) with multiple-level lesions. L5 spondylolysis affected 432 vertebrae and was the most common location (73.8%), followed by 125 L4 lesions (21.4%), 24 L3 lesions (4.1%), and 2 L2 lesions (0.7%). CONCLUSIONS: The percentage of L4 lesions in our study was significantly higher and of L5 lesions was significantly lower than those lesions' percentages in the population-based study. L4 spondylolysis may be more unstable or cause clinical symptoms more frequently leading to more surgical intervention. The percentage of multiple-level spondylolysis was similar between the 2 studies, suggesting these patients respond relatively well to conservative treatment. The male/female ratio was 2:1 in both studies, indicating that males and females require surgery at a similar frequency.

A Fatal Case of Relapsing Pneumonia Caused by Legionella pneumophila in a Patient with Rheumatoid Arthritis After Two Injections of Adalimumab.

We present a rare fatal case of relapsing pneumonia caused by Legionella pneumophila in a patient with rheumatoid arthritis after only two injections of adalimumab. A 78-year-old Japanese woman with a 14-year history of rheumatoid arthritis was prescribed adalimumab because her disease activity remained high. However, 8 days after her second injection of adalimumab, she was admitted to our hospital and diagnosed with pneumonia caused by L. pneumophila. Following intravenous antibiotic therapy, she recovered completely from pneumonia and was discharged on day 10, but pneumonia relapsed, resulting in death 79 days after the first episode of pneumonia. L. pneumophila can lead to recurrence of pneumonia that can ultimately prove fatal, similar to the present case. A review of the pertinent literature is also presented.

The effect of a prostaglandin E1 derivative on the symptoms and quality of life of patients with lumbar spinal stenosis.

BACKGROUND: Quality of life (QOL) is a concern for patients with lumbar spinal stenosis (LSS). In this study, QOL was examined using the 5-item EuroQol (EQ-5D). METHODS: QOL and activities of daily living (ADL) were surveyed for 91 patients who visited 18 medical institutions in our prefecture and were diagnosed with LSS-associated intermittent claudication. A second survey was performed after ≥6 weeks for 79 of the subjects to evaluate therapy with limaprost (an oral prostaglandin E1 derivative) or etodolac (an NSAID). Symptoms, maximum walking time, QOL, ADL items, and relationships among these variables were investigated for all 91 patients. Leg pain, leg numbness, and low back pain while walking were surveyed by use of VAS scores (0-100). RESULTS: Leg pain, leg numbness, and low back pain while walking (VAS ≥25) were present in 83.5, 62.6, and 54.9 % of the patients in the first survey, and approximately half of the patients had a maximum walking time <15 min. The mean EQ-5D utility value for QOL was 0.59 ± 0.12. This value was significantly associated with maximum walking time (p = 0.030) based on classification of patients into groups with walking times <7.5, 7.5-15, 15-30, and >30 min, showing that maximum walking time affected health-related QOL. Of the 79 patients who completed the second survey, 56 had taken limaprost and 23 (control group) had received etodolac. Limaprost improved possible walking time, reduced ADL interference, and significantly increased the EQ-5D utility score, whereas no significant changes occurred in the control group. Maximum walking time was prolonged by ≥10 min and the EQ-5D utility value was improved by ≥0.1 points in significantly more patients in the limaprost group than in the control group. CONCLUSION: According to the findings of this survey, at an average of 8 weeks after administration limaprost improved symptoms, QOL, and ADL in LSS patients whereas treatment with an NSAID reduced pain but did not have any other effects.

Primary spinal cord tumors: review of 678 surgically treated patients in Japan. A multicenter study.

PURPOSE: To clarify the relative frequency of various histopathological primary spinal cord tumors and their features in Japanese people and to compare this data with other reports. METHODS: Primary spinal cord tumor surgical cases from 2000 to 2009, which were registered in our affiliated hospital database were collected. We examined age at surgery, sex, anatomical location, vertebral level of the tumor, and pathological diagnosis in each case. RESULTS: Of the 678 patients in our study, 377 patients (55.6 %) were males and 301 patients (44.4 %) were females (male/female ratio 1.25). The mean age at surgery was 52.4 years. Of these tumors, 123 cases (18.1 %) were intramedullary, 371 cases (54.7 %) were intradural extramedullary, 28 cases (4.1 %) were epidural, and 155 cases (22.9 %) were dumbbell tumors. The pathological diagnoses included 388 schwannomas (57.2 %), 79 meningiomas (11.6 %), 54 ependymomas (8.0 %), 27 hemangiomas (4.0 %), 23 hemangioblastomas (3.4 %), 23 neurofibromas (3.4 %), and 9 astrocytomas (1.3 %). The male/female ratios for schwannomas, meningiomas, ependymomas, hemangiomas, hemangioblastomas, neurofibromas, malignant lymphomas, and lipomas are 1.4, 0.34, 1.3, 1.5, 2.3, 1.3, 2.7 and 2.3, respectively. CONCLUSION: This is the first published research in English on the epidemiology of primary spinal cord tumors in Japanese people. Similar to other reports from Asian countries, our data indicates a higher male/female ratio overall for spinal cord tumors, a higher proportion of nerve sheath cell tumors, and a lower proportion of meningiomas and neuroepithelial tumors compared to reports from non-Asian countries. Data in the current study represent the characteristics of primary spinal cord tumors in Asian countries.

Pneumococcal polyarticular septic arthritis after a single infusion of infliximab in a rheumatoid arthritis patient: a case report.

INTRODUCTION: We present a case of Streptococcus pneumoniae polyarticular septic arthritis in a patient with rheumatoid arthritis receiving a single infusion of infliximab. CASE PRESENTATION: A 38-year-old Japanese man with a 5-year history of seronegative rheumatoid arthritis had previously received sulphasalazine and methotrexate therapies and was on regular low-dose prednisolone therapy. Despite these treatments, his disease activity remained high and infliximab was introduced in addition to methotrexate, prednisolone, and folic acid. However, he was admitted to hospital with a fever of 40.6°C, chills, and polyarthralgia eight days after the first infusion of infliximab. His joints were swollen, painful, and warm. Laboratory data showed marked acute inflammation. He was diagnosed with bacterial septic polyarthritis, and emergency surgical joint lavage and drainage was performed at the knees along with needle aspiration and lavage of the ankles and right wrist. He was then given intravenous antibiotic therapy for 31 days. He made a good recovery and was discharged on day 37. CONCLUSIONS: We believe this is the first reported case of severe pneumococcal septic arthritis requiring hospitalization in a patient treated with infliximab. S. pneumonia is now a well-recognized but uncommon cause of polyarticular septic arthritis that can lead to cessation of therapy, as in our patient's case.

An arterial pulse examination is not sufficient for diagnosis of peripheral arterial disease in lumbar spinal canal stenosis: a prospective multicenter study.

STUDY DESIGN: Prospective, multicenter study. OBJECTIVE: To conduct peripheral arterial disease (PAD) screening on intermittent claudication (IC) in patients with lumbar spinal canal stenosis (LSCS) to examine the relationships among combined LSCS and PAD, symptoms, and physical findings. SUMMARY OF BACKGROUND DATA: IC occurs due to two underlying diseases, LSCS and PAD, and has an increasing prevalence with the aging of society. Reliable diagnosis of PAD is critical for appropriate conservative management of IC patients with LSCS in an Orthopedic Surgery Outpatient Department (OSOPD). METHODS: PAD tests were prospectively conducted in 201 patients with IC and LSCS who initially visited an OSOPD at a hospital affiliated with the Nogoya Spine Group. Occurrence of PAD as a complication was assessed using ankle brachial pressure index (ABI) and toe brachial pressure index (TBI) tests. PAD was diagnosed in patients with ABI ≤ 0.9 or TBI ≤ 0.6, and the relationship of the occurrence of PAD with symptoms and physical findings such as abnormal arterial pulses was investigated. RESULTS: Combined LSCS and PAD was found in 52 patients (26%), with 45 cases (22%) diagnosed on the basis of TBI test in patients with a normal ABI. Of the patients with PAD, many suffered from risk factors for PAD, with a significantly higher frequency of PAD in patients with hyperlipidemia (P < 0.05). PAD also occurred significantly more frequently in patients with abnormal pulses in the popliteal (P < 0.05), posterior tibial (P < 0.0001), and dorsal pedis (P < 0.0001) arteries; however, the sensitivity of these tests for PAD diagnosis was relatively low, at 34%, 60% and 68%, respectively. CONCLUSION: The results of the prospective study define the rate of occurrence of combined LSCS and PAD using ABI and TBI tests for the first time, and the findings suggest that screening for PAD should be conducted in LSCS patients. ABI and TBI tests are necessary for PAD screening in outpatients, whereas observation of the arterial pulse in the lower extremities is necessary but not sufficient for PAD diagnosis.

Retrospective multicenter study of surgical treatments for osteoporotic vertebral fractures.

BACKGROUND: Although many surgical procedures are available for treating osteoporotic vertebral fractures, there have been no comprehensive multicenter surveys in Japan focusing on surgical treatments for these fractures. This study aimed at (1) conducting a retrospective multicenter study to survey surgical treatments performed at referral center hospitals in various regions in Japan and (2) analyzing situations and problems related to the surgical treatments of osteoporotic vertebral fractures in Japanese hospitals. METHODS: Among 738 patients who were hospitalized in 13 hospitals in various regions in Japan between 2005 and 2006 for osteoporotic vertebral fractures, 84 patients (11.4%) who underwent spinal surgery were enrolled. These patients were retrospectively analyzed regarding cause of injury, preoperative symptoms, preoperative neurological function, surgical procedures, periods of bed rest, length of hospital stay, and ambulatory status at discharge from hospital. RESULTS: As to the cause of spinal fracture, 38 patients (45% of the surgical patients) could not identify a specific cause of their spinal fracture. Preoperative neurological motor weakness in legs was observed in 41 (49%). With regard to surgical treatment, posterior spinal reconstruction surgery was performed in 50 patients (60%), vertebroplasty in 26 (31%), anterior reconstruction surgery in 6 (7%), anterior and posterior combined reconstruction surgery in 1, and posterior decompression alone in 1 patient. In all, 70 patients (83.3%), whose periods of hospital stay averaged 52.8 days, could walk by themselves at the time of discharge; 14 (16.7%), whose periods of hospital stay averaged 44.7 days, could not walk by themselves at the time of discharge. CONCLUSIONS: Even after a large variety of surgical procedures were tried to treat osteoporotic vertebral fractures and long hospital stays, about 17% of the patients were unable to walk by themselves at the time of discharge from hospital.

Nationwide survey of current medical practices for hospitalized elderly with spine fractures in Japan.

BACKGROUND: The status of hip fracture incidence and treatment is well known through nationwide surveys in Japan. However, there have been no similar studies on spine fractures. Therefore, we investigated current medical practices for them. METHODS: Altogether, 1200 hospitals were randomly selected for the survey with consideration of region and hospital characteristics. Questionnaire items included the number of hospitalized spine patients, imaging test implementation, type of conservative treatment, use of open surgery and vertebroplasty, and the number of these procedures performed in 2005. RESULTS: Responses were received from 473 hospitals. On the day of response, there were 14 372 hospitalized orthopedic patients (average 32.8/hospital). Among them were 1403 spine fracture patients (3.1/hospital), accounting for 13.5% of orthopedic patients. Of them, 91.9% received conservative treatment. The mean percentage of spine fracture patients who were hospitalized was 39.5%. The most reliable imaging test was said to be magnetic resonance imaging. Casting or bracing was used in most of the institutions. The most common analgesic treatment was oral nonsteroidal antiinflammatory drugs. Open surgery and vertebroplasty were conducted for spine fractures in the elderly at 26.5% and 16.3% of hospitals, respectively. In these hospitals, 624 and 257 patients underwent open surgery and vertebroplasty, respectively, in 2005. CONCLUSIONS: In Japan, more than 90% of elderly patients hospitalized with spine fractures received conservative treatment. Surgical treatment, either open surgery or vertebroplasty, was performed at 30% of the hospitals. This study provides basic data that will contribute to planning improvements in spinal fracture treatment in the elderly.

The migration of a broken luque rod: a case report.

An exceedingly rare complication of Luque segmental spinal instrumentation in spinal fractures is described. A patient was treated for fractures of the eighth and ninth thoracic vertebra associated with traumatic paraplegia using Luque segmental spinal instrumentation. Ten years postoperatively, broken rods and sublaminar wires were found. One of the broken rods migrated caudad penetrating the sacrum and protruding into the pelvic cavity. The rod had projected into the rectum, and was extracted through the wall of the rectum and the anus. This case report emphasizes the importance of careful surgical technique and long-term follow up for patients who had undergone spinal instrumentation surgery.

Intra-articular synovial lipoma of the knee joint (located in the lateral recess): a case report and review of the literature.

This report describes a rare intra-articular synovial lipoma of the knee joint which developed in a 66-year-old female. The patient suffered from sudden knee pain and a catching or slight interruption of normal motion of the left knee and then noticed the tumor. The tumor was located in the lateral recess of the knee joint and showed a signal intensity similar to subcutaneous fat on T1 and T2()-weighted magnetic resonance images. The arthroscopy revealed a smooth, globular, yellowish, encapsulated tumor extending into the lateral recess from the surface of the lateral condyle and the tumor was totally excised under arthroscopic guidance. Histologic examination of the specimen revealed a tumor composed of mature adipose cells covered by a thin fibrous layer of varying thickness and normal synovial lining cells. The diagnosis was intra-articular synovial lipoma. Intra-articular synovial lipomas should be distinguished from other similar lipomatoid conditions such as Hoffa disease and villous lipomatous proliferation of the synovial membrane (lipoma arborescens). Intra-articular synovial lipoma should be considered in the differential diagnoses when examining a patient with sudden knee pain, and a catching or locking knee.

Catechol-O-methyltransferase gene polymorphisms in benign prostatic hyperplasia and sporadic prostate cancer.

Various carcinogenic metabolites, including catechol estrogens, play a role in malignant transformation. An enzyme that is capable of neutralizing the genotoxic effects of these compounds is catechol-O-methyltransferase (COMT). A variant form of this enzyme has been shown to reduce its activity by up to 4-fold; thus, we hypothesize that single nucleotide polymorphisms of the COMT gene can be a risk factor for benign prostatic hyperplasia (BPH) and prostate cancer. To test this hypothesis, the genetic distribution of three different COMT polymorphisms at codon 62 (C-->T), codon 72 (G-->T), and codon 158 (G-->A) were analyzed in 131 normal healthy subjects, 134 BPH, and 178 sporadic prostate cancer samples from a Japanese population. Results of these experiments show that the variant genotype at codon 62 (P = 0.060) and codon 158 (P = 0.047) are risk factors for prostate cancer but not BPH when compared with normal controls. Odds ratio (OR) and 95% confidence interval (95% CI) for cancer were 3.24 and 1.38 to 7.61, respectively, for codon 62 T/T genotype when compared with wild type. At codon 158, the A/A variant for cancer had an OR of 3.00 with a 95% CI of 1.38 to 6.54 compared with wild type. Codons 62 and 158 were in linkage disequilibrium (LD), and when compared with the C-G haplotype, other types (C-A, T-G, T-A) were observed to be associated with prostate cancer (P = 0.040) but not BPH. Codon 72 on the other hand, was not in LD with either codon 62 or 158. The homozygous variant on codon 72 was rare in this Japanese population, and the heterozygous G/T at this codon was not associated with either prostate cancer or BPH. When evaluating the risk of COMT polymorphisms with stage or grade of cancer, no associations were observed for any of the genotypes with the exception of a tendency (P = 0.096) for the variant A allele on codon 158 to be correlated with higher stages (> or = T3) of cancer. This is the first report that shows the polymorphisms of COMT to be associated with sporadic prostatic carcinogenesis. These results are important in understanding the role of COMT polymorphisms in the pathogenesis of prostate cancer.

Functional Loss of the gamma-catenin gene through epigenetic and genetic pathways in human prostate cancer.

Gamma-catenin is a cell adhesion molecule and a candidate mediator of Wnt signal transduction. We hypothesized that impaired regulation of gamma-catenin through genetic and epigenetic pathways is associated with the pathogenesis of prostate cancer. To test this hypothesis, cytosine-phosphate-guanine methylation, loss of heterozygosity (LOH), and mutation status of the gamma-catenin gene were analyzed in cultured prostate cancer cell lines, 180 localized prostate cancers, 69 benign prostatic hyperplasias, and 11 hormone refractory prostate cancers (HRPC). In prostate cancer cell lines (DuPro, LNCaP, ND-1, and PC3), gamma-catenin mRNA transcripts were increased after 5-aza-2'-deoxycytidine treatment. In localized prostate cancer, gamma-catenin expression was lower but prevalence of gamma-catenin methylation was higher compared with benign prostatic hyperplasia. However, gamma-catenin methylation did not correlate with Gleason sum, pT category, or capsular penetration. Among localized prostate cancers with positive gamma-catenin methylation, the presence of LOH at chromosome 17q21 was closely related to down-regulation of gamma-catenin mRNA expression. The gamma-catenin mutations were not found in localized prostate cancers, whereas six mutations were found in five HRPCs within or close to the GSK-3beta consensus motif phosphorylation site, among which four HRPCs showed strong nuclear gamma-catenin accumulation. In these four HRPCs, Bcl-2 expression was increased, whereas the target of the Wnt signal, c-myc, was only expressed in one HRPC. Therefore, although epigenetic gamma-catenin methylation is an early event in the development of prostate cancer, simultaneous events of epigenetic cytosine-phosphate-guanine methylation and genetic LOH may be responsible for functional loss of gamma-catenin. The gamma-catenin mutation related to Bcl-2 overexpression has a significant effect on the pathogenesis of HRPC. This is the first report to characterize the epigenetic and genetic regulation of gamma-catenin in human prostate cancer.

Methylation of the gamma-catenin gene is associated with poor prognosis of renal cell carcinoma.

PURPOSE: Gamma-catenin is a cell adhesion protein, and its functional loss is associated with tumor invasion and metastasis. We hypothesize that (1) promoter CpG methylation regulates the expression and function of the gamma-catenin gene in renal cell carcinoma (RCC) and (2) methylation of the gamma-catenin gene is associated with poor prognosis of RCC. To test these hypotheses, we analyzed the CpG methylation status of the gamma-catenin gene and its correlation with clinical outcome in RCC. EXPERIMENTAL DESIGN: Genomic DNA and total RNA were extracted from three renal cancer cell lines (A498, Caki-1, and Caki-2) and 54 RCC tissue samples with their corresponding normal kidney tissue samples. Expression of gamma-catenin gene was analyzed by reverse transcription-PCR and immunostaining. Promoter methylation was analyzed by two different methylation-specific PCR (MSP-A and MSP-B), and the results were verified by DNA sequencing. RESULTS: The demethylating agent (5-aza-2'-deoxycytidine) increased levels of mRNA transcript of the gamma-catenin gene in three renal cancer cell lines. Gamma-catenin mRNA and protein expression were significantly reduced in RCC samples compared with normal kidney samples, respectively (P < 0.05). MSP-A and MSP-B bands were detected in 45 of 54 (83.3%) and 49 of 54 (90.7%) RCC samples, respectively. In normal kidney, weak products of MSP-A and MSP-B were detected in 5 of 54 (9.3%) and 6 of 54 (11.1%) samples, respectively. Likewise, both MSP-A and MSP-B ratios were significantly higher in RCC samples compared with normal kidney samples, respectively (P < 0.01). Multivariate analysis revealed that the MSP-B ratio was a powerful and independent predictor superior to nuclear grade and Robson stage with respect to survival and disease progression (P = 0.029 and 0.0071, respectively). No mutations in the NH(2)-terminal region of gamma-catenin were found in this study. CONCLUSION: Expression of gamma-catenin is regulated by promoter CpG methylation, and the balance between methylated and unmethylated RCC cell populations could determine its functional role. Because the conventional nuclear grade and/or staging system have some limitations to predict precise clinical outcome, this is the first report demonstrating that promoter CpG methylation of gamma-catenin can be an independent and superior predictor for survival and disease progression.

Inactivation of the hMSH3 mismatch repair gene in bladder cancer.

Deficiency in the DNA mismatch repair (MMR) is frequently involved in various cancers. The hMSH3 gene is one of the human MMR genes whose role in bladder cancer is not known. We hypothesized that down-regulation of the hMSH3 gene might be involved in bladder cancer. In this study we analyzed this gene with regard to frame-shift mutation, single nucleotide polymorphism (SNP), a 9bp repeat in exon 1, loss of heterozygosity (LOH), immunohistochemistry, and methylation status in 102 bladder cancer samples. Immunohistochemistry revealed that hMSH3 expression in bladder cancer was significant decreased compared to normal epithelium (p<0.0001). An inverse correlation with pathological grade was found. The frame-shift mutation in the (A) 8 tract was lacking in bladder cancer. There was no significantly difference between bladder cancer samples and healthy controls' with regard to SNP and the 9bp repeat. In bladder cancer, presence of the codon 222 polymorphism, LOH, and the 9bp repeats in exon 1 had a correlation with either pathological stage or pathological grade. Presence of the codon 1036 polymorphism had significant correlation with pathological stage and a trend to correlation with pathological grade. After 5-aza-dC treatment, MSH3 expression was significantly enhanced in TCC and UMUC bladder cancer cells when compared to untreated cells. This is the first report suggesting that genetic and epigenetic alterations in the human MSH3 gene might play a significant role in the progression of bladder tumors.

Age-dependent methylation of ESR1 gene in prostate cancer.

The incidence of prostate cancer increases dramatically with age and the mechanism underlying this association is unclear. Age-dependent methylation of estrogen receptor alpha (ESR1) gene has been previously implicated in other cancerous and benign diseases. We evaluated the age-dependent methylation of ESR1 in prostate cancer. The methylation status of ESR1 in 83 prostate cancer samples from patients aged 49 to 77 years (mean age at 67.4 years) was examined using the bisulfite genomic sequencing technique. The samples were divided into three age groups: men aged 60 years and under (n = 14), men aged 61-70 years (n = 40), and men aged over 70 years (n = 29). Overall, ESR1 promoter methylation was detected in 54 out of 83 (65.1%) prostate samples. The methylation rate of ESR1 increased dramatically with age from 50.0% in patients aged 60 years and under to 89.7% for patients aged 70 years and over. Logistic regression analyses revealed that age and Gleason score were the only variables that affect incidence of ESR1 methylation; other clinical factors such as prostate-specific antigen level and clinical stage did not. We also calculated ESR1 methylation density (the percentage of methylated CpGs among all CpGs within the analyzed region) and severity (the percentage of methylated CpG alleles) for each sample analyzed. Multiple regression analyses showed a positive correlation between age and methylation density (beta, 0.35; P, 0.012; 95% CI, 0.26-2.01); while Gleason score was positively associated with methylation severity (beta, 0.45; P, 0.018; 95% CI, 1.04-4.26). These findings suggest that methylation of ESR1 is both age-dependent and tumor differentiation-dependent and age-dependent methylation of ESR1 may represent a mechanism linking aging and prostate cancer.