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Contemporary treatment of pediatric acute myeloid leukemia (AML) requires the assignment of patients to specific risk groups. To explore whether expression profiling of leukemic blasts could accurately distinguish between the known risk groups of AML, we analyzed 130 pediatric and 20 adult AML diagnostic bone marrow or peripheral blood samples using the Affymetrix U133A microarray. Class discriminating genes were identified for each of the major prognostic subtypes of pediatric AML, including t(15;17)[PML-RARalpha], t(8;21)[AML1-ETO], inv(16) [CBFbeta-MYH11], MLL chimeric fusion genes, and cases classified as FAB-M7. When subsets of these genes were used in supervised learning algorithms, an overall classification accuracy of more than 93% was achieved. Moreover, we were able to use the expression signatures generated from the pediatric samples to accurately classify adult de novo AMLs with the same genetic lesions. The class discriminating genes also provided novel insights into the molecular pathobiology of these leukemias. Finally, using a combined pediatric data set of 130 AMLs and 137 acute lymphoblastic leukemias, we identified an expression signature for cases with MLL chimeric fusion genes irrespective of lineage. Surprisingly, AMLs containing partial tandem duplications of MLL failed to cluster with MLL chimeric fusion gene cases, suggesting a significant difference in their underlying mechanism of transformation.

A simple two-step method was developed for the covalent immobilization of atom-transfer radical polymerization (ATRP) initiators on the hydrogen-terminated Si(100) (Si-H) surface. Well-defined functional polymer-Si hybrids, consisting of covalently tethered brushes of poly(ethylene glycol) monomethacrylate (PEGMA) polymer, N-isopropylacrylamide (NIPAAm) polymer, and NIPAAm-PEGMA copolymers and block copolymers on Si-H surfaces, were prepared via surface-initiated ATRP. Kinetics study revealed that the chain growth from the silicon surface was consistent with a "controlled" process. Surface cultures of the cell line 3T3-Swiss albino on the hybrids were evaluated. The PEGMA graft-polymerized silicon [Si-g-P(PEGMA)] surface is very effective in preventing cell attachment and growth. At 37 degrees C [above the lower critical solution temperature (LCST, approximately 32 degrees C) of NIPAAm], the seeded cells adhered, spread, and proliferated on the NIPAAm graft polymerized silicon [Si-g-P(NIPAAm)] surface. Below the LCST, the cells detached from the Si-g-P(NIPAAm) surface spontaneously. Incorporation of PEGMA units into the NIPAAm chains of the Si-g-P(NIPAAm) surface via copolymerization resulted in more rapid cell detachment during the temperature transition. The "active" chain ends on the Si-g-P(PEGMA) and Si-g-P(NIPAAm) hybrids were also used as the macroinitiators for the synthesis of diblock copolymer brushes. Thus, not only are the hybrids potentially useful as stimuli-responsive adhesion modifiers for cells in silicon-based biomedical microdevices but also the active chain ends on the hybrid surfaces offer opportunities for further surface functionalization and molecular design.

Nasal NK/T cell lymphoma is a distinctive type of extranodal lymphoma with an unique immunophenotype and a strong association with Epstein-Barr virus (EBV). It is one of the common extranodal lymphomas in Taiwan. We studied 22 cases of nasal NK/T cell lymphoma to characterize their clinicopathologic features and to explore the possible differences between histologic subtypes and their clinical behavior as well as the prevalence of 30-base pair (bp) deleted latent membrane protein-1 (LMP-1) gene of the EBV. They consisted of 5 cases of small cell type (SC), 6 cases of medium-sized cell type (MC), 6 cases of large cell type (LC), and 5 cases of pleomorphic cell type (PC). Twelve patients were men and 10 were women (1.2 to 1), and their ages ranged from 34 to 75 years with a median age of 55.5 years. The median ages of the LC type and PC type were older than the other 2 types. No other clinical features differed significantly among the 4 subtypes. Nasal obstruction was the most common initial presenting symptom. All but 1 case had stage IE disease at the time of diagnosis. Five cases developed extranasal involvement and skin was the most common site. No bone marrow involvement was detected. The majority of patients received local radiotherapy and chemotherapy. Local irradiation was more effective than chemotherapy alone. We achieved an overall survival of 63.6% at 5 years as estimated by the Kaplan-Meier analysis, which was better than other series. All cases displayed an immunophenotypic profile of CD3(epsilon)+, CD20-, CD56+, and TIA-1+ except that 1 case was CD3(epsilon)-. Fourteen of 22 cases (64%) expressed LMP-1. Nine cases of various cell types (41%) were also CD30+. Among the 4 histologic subtypes, the SC type differed from the other types by the absence of angiodestruction and necrosis, although angioinvasive growth was seen in 2 of them. Pseudoepitheliomatous hyperplasia was seen in only 3 cases of the SC type, and all 5 cases of the SC type were CD30-. No statistical difference in survival was found among the 4 histologic subtypes or between CD30+ and CD30- cases. All 22 cases were positive for EBV by polymerase chain reaction and Epstein-Barr virus early RNA (EBER) in-situ hybridization. A high prevalence rate of 86% (19/22) of the 30-base pair (bp) deleted LMP-1 gene was found, but 81.5% (22/27) of the EBV-positive control reactive lymphoid tissues also had the 30-bp deleted LMP-1 gene. Therefore, the high prevalence of the 30-bp deleted LMP-1 gene found in NK/T cell lymphoma could be due to the high prevalence of the deleted variant in this geographic region. However, it remains possible that the high prevalence of the deleted LMP-1 gene contributed to the increased incidence of EBV-associated nasal NK/T cell lymphoma in Taiwan.

Background: The prognostic significance of internal tandem duplication (ITD) of the fms-like tyrosine kinase 3 gene (FLT3) for patients with myelodysplastic syndrome (MDS) is not clearly defined. In the current study, the authors sought to assess the value of FLT3/ITD mutation status as a prognostic genetic marker for patients with MDS., Methods: FLT3/ITD mutation status was evaluated by performing DNA polymerase chain reaction assays on 198 bone marrow samples obtained from patients with MDS at initial diagnosis. All aberrant products were sequenced, and GeneScan analysis was performed to measure FLT3/ITD mutant levels., Results: Five patients (2.5%)--2 of the 99 patients who had refractory anemia with excess blasts and 3 of the 51 patients who had chronic myelomonocytic leukemia--had FLT3/ITD mutations. FLT3/ITD was not observed in any of the 48 patients who had refractory anemia (with or without ringed sideroblasts). There was no significant difference in clinicohematologic characteristics, cytogenetic characteristics, or International Prognostic Scoring System score between the FLT3/ITD-positive group and the FLT3/ITD-negative group. Four of the 5 patients carrying the FLT3/ITD mutation experienced progression of disease to acute myeloid leukemia (AML), compared with 70 of the 193 patients who did not have FLT3/ITD (P = 0.066). In addition, progression to AML was more rapid in patients with FLT3/ITD-positive disease than in patients with FLT3/ITD-negative disease (mean +/- standard error, 3.0 +/- 0.5 months vs. 62.8 +/- 5.6 months; P < 0.0001). Patients with FLT3/ITD-positive disease also had significantly shorter survival compared with patients who had FLT3/ITD-negative disease (mean +/- standard error, 5.2 +/- 1.4 months vs. 33.7 +/- 3.1 months; P < 0.0001). On multivariate analysis, FLT3/ITD was identified as an independent predictor of reduced time to development of AML (P = 0.0001) and reduced overall survival (P = 0.002)., Conclusions: The results of the current study demonstrate that FLT3/ITD is associated with a high risk of transformation to AML, rapid progression of AML, and poor survival in patients with MDS., (Copyright 2004 American Cancer Society.)

Objective: Several investigators have speculated that mechanical stress might play an important role in the development of hand osteoarthritis (OA). Chopsticks, used universally as eating utensils in China, increase joint loading in the first through third fingers. We conducted a population-based survey among elderly Chinese individuals living in Beijing, to explore whether chopsticks use is associated with prevalent hand OA., Methods: We recruited a sample of persons ages 60 years and older, using door-to-door enumeration in randomly selected neighborhoods in Beijing. Subjects answered questions about the hand with which they use chopsticks, handedness, and pincer grip activities. Bilateral posteroanterior hand radiographs were obtained, and each joint was graded according to the Kellgren/Lawrence (K/L) scale. We defined a subject as having radiographic OA if at least 1 of his or her hand joints had radiographic OA (K/L score of > or =2). We defined a particular hand group (i.e., distal interphalangeal [DIP] joints, proximal interphalangeal [PIP] joints, or metacarpophalangeal [MCP] joints) as having OA if at least 1 joint of the group had radiographic OA. We calculated the prevalence of OA for each hand joint and, according to the status of chopsticks use, performed a matched analysis to examine the relationship between chopsticks use and the prevalence of hand OA. In the analysis, we excluded persons who reported a previous hand injury. Because most subjects used chopsticks with their dominant hand (a hand they would be expected to use more for all manual tasks), we also performed the analysis among subjects who reported that they had no hand preference when performing other activities and subjects who denied other pincer grip activities., Results: A total of 1,008 men and 1,499 women were assessed. The prevalence ratio for OA of the thumb IP joint in the chopsticks hand was 1.2 (range 1.1-1.4) in men and 1.6 (range 1.4-1.7) in women; the prevalence ratio for OA of the second and third PIP joints was 1.5 (range 1.1-2.2) in men and 1.4 (range 1.2-1.7) in women; and the prevalence ratio for OA of the second and third MCP joints was 1.4 (range 1.2-1.6) in men and 1.4 (range 1.2-1.6) in women. The prevalence ratios in these joints were greater than the ratios in other MCP, PIP, or DIP joints from the same hand that were unlikely to be involved by chopsticks use, especially among women. Similar results were observed when the analyses were limited to ambidextrous subjects and subjects who did not engage in any other pincer grip activities. Thumb IP joint OA affected 26% of the entire population studied, and chopsticks use accounted for 19% of the risk of OA developing in this joint in men and 36% of the risk in women., Conclusion: This epidemiologic study investigated the relationship of chopsticks use to hand arthropathy. The results suggest that chopsticks use is associated with an increased prevalence of OA in the IP joint of the thumb, and in the second and third PIP and MCP joints.

Objective: To examine the association between squatting, a common daily posture in China, and the prevalence of radiographic osteoarthritis (OA) in different knee compartments among Chinese subjects from Beijing, and to estimate how much of the difference in prevalence of knee OA between Chinese subjects in Beijing and white subjects in Framingham, Massachusetts is accounted for by the impact of squatting., Methods: We recruited a random sample of Beijing residents age > or =60 years. Subjects answered questions on joint symptoms, and knee radiographs were obtained. Subjects were also asked to recall the average amount of time spent squatting each day at age 25 years. Radiographic films (weight-bearing anteroposterior and skyline views) were read for Kellgren/Lawrence (K/L) grade and individual radiographic features. Medial disease was defined when radiographs showed a K/L grade of > or =2 at the tibiofemoral joint and a medial joint space narrowing score of > or =1, and lateral disease was assessed in a comparable manner in the lateral compartments. We examined the association of squatting with the prevalence of tibiofemoral OA as well as with the prevalence of patellofemoral knee OA, while adjusting for age and other potential confounding factors. We used the same approach to assess the relationship between squatting and tibiofemoral OA in the medial compartment and in the lateral compartment. Finally, we estimated the impact of squatting at age 25 on the difference in prevalence of knee OA between Chinese subjects in Beijing and white subjects in the Framingham OA Study., Results: Squatting was very common among the Chinese subjects: approximately 40% of men and approximately 68% of women reported squatting > or =1 hour per day at age 25. The prevalence of tibiofemoral OA increased as the time spent squatting at age 25 increased in both the men and the women. Compared with subjects who squatted <30 minutes per day at age 25, the multivariable-adjusted prevalence odds ratios of tibiofemoral OA were 1.1 for time spent squatting of 30-59 minutes/day, 1.0 for 60-119 minutes/day, 1.7 for 120-179 minutes/day, and 2.0 for > or =120 minutes/day among the men (P for trend = 0.074), and the respective odds ratios among the women were 1.4, 1.3, 1.2, and 2.4 (P for trend = 0.077). A weaker association with patellofemoral OA was found. Prolonged squatting in daily life was more strongly associated with medial knee OA than with lateral disease in the men, but had a similar effect on both knee compartments in the women. After adjusting for the impact of squatting, the age-adjusted difference in prevalence of tibiofemoral OA was reduced from an excess of 14.4% to 9.5% in the Chinese women, but the difference in prevalence of tibiofemoral OA in the Chinese men increased after adjustment for age and squatting, from 2.9% lower to 7.0% lower as compared with their white counterparts., Conclusion: Prolonged squatting is a strong risk factor for tibiofemoral knee OA among elderly Chinese subjects in Beijing, and accounts for a substantial proportion of the difference in prevalence of tibiofemoral OA between Chinese subjects in Beijing and white subjects in Framingham.

Chemokines regulate the chemotaxis, development, and differentiation of many cell types enabling the regulation of routine immunosurveillance and immunological adaptation. CC chemokine receptor 1 (CCR1) is the target of 11 chemokines. This promiscuity of receptor-ligand interactions and the potential for functional redundancy has led us to investigate the selective activation of CCR1-coupled pathways by known CCR1 agonists. Chemokines leukotactin-1, macrophage inflammatory protein (MIP)-1alpha, monocyte chemotactic peptide (MCP)-3, RANTES, and MIP-1delta all inhibited adenylyl cyclase activity in cells transiently transfected with CCR1. In contrast, only MIP-1delta was unable to signal via G14-, G16- or chimeric 16z44-coupled pathways. In a stable cell line expressing CCR1 and Galpha14, all of these five chemokines along with hemofiltrate CC chemokine (HCC)-1 and myeloid progenitor inhibitory factor (MPIF)-1 were able to stimulate G(i/o)-coupled pathways, but MIP-1delta, HCC-1 and MPIF-1 were unable to activate G14-mediated stimulation of phospholipase Cbeta activity. In addition, MIP-1delta was unable to promote the phosphorylation of extracellular signal-regulated kinase and c-Jun N-terminal kinase. This suggests that different chemokines are able to selectively activate CCR1-coupled pathways, probably because of different intrinsic ligand efficacies. CCR1 and Galpha14 or Galpha16 are co-expressed in several cell types and we hypothesize that selective activation of chemokine receptors provides a mechanism by which chemokines are able to fine-tune intracellular signaling pathways.

Purpose: We analyzed Asp(835) mutations of FLT3 on paired marrow samples at diagnosis and relapse from 120 adult patients with de novo acute myeloid leukemia (AML) to determine the role of FLT3 Asp(835) mutation in the relapse of AML., Experimental Design: Asp(835) mutation was analyzed by DNA PCR amplification of exon 20 of FLT3 gene followed by EcoRV digestion. All of the mutations were confirmed by sequence analysis. Mutant to wild-type allelic ratio was determined by Genescan analysis. The Expand Long Template PCR System was used to determine the allelic location of internal tandem duplication of FLT3 (FLT3/ITD) and Asp(835) mutations., Results: Thirteen patients had Asp(835) mutations at diagnosis, of them 8 lost the mutations at relapse, and the remaining 5 patients carrying Asp(835) mutations at diagnosis relapsed with the identical mutation types. Another 6 patients acquired Asp(835) mutations at relapse. Five samples harbored both FLT3/ITD and Asp(835) mutations that were found on different alleles by cloning analysis in the 3 patients studied. There were no differences in WBC count, French-American-British subtype, percentage of marrow blasts, or circulating blasts between patients with and without Asp(835) mutations, whereas the difference in the prevalence of Asp(835) mutations among cytogenetic/molecular subgroups was statistically significant (P = 0.025)., Conclusions: The present study showed that patients with AML had heterogeneous patterns of FLT3 Asp(835) mutations, either acquisition or loss of the mutations at relapse. Asp(835) mutant clone may develop as a secondary event in a subset of patients with AML.

Twenty-three patients with primary salivary gland lymphoma were diagnosed between 1990 and 2001. The sites of involvement were the parotid gland in 13, the submandibular gland in 9 and the minor salivary gland in 1. The sites of lymphoma involvement beyond the salivary glands were the cervical lymph nodes in 7, bone marrow in 3, the axillary lymph nodes in 3, the nasopharynx in 2, the abdominal lymph nodes in 2, the palate, the subconjunctiva, and the spleen in 1 each patient. Histologically, 19 patients had lymphomas of mucosa-associated lymphoid tissue (MALT) with myoepithelial sialadenitis in 13, 3 patients had diffuse large cell lymphomas and 1 had follicular lymphoma. Six patients were in stage I, 4 in II, 1 in III and 12 in IV. Eight of 23 patients (35%) had autoimmune diseases before or after the diagnosis of NHL and all suffered from MALT lymphoma. Four patients with parotid MALT lymphoma had primary or secondary Sjogren's syndrome. One each patient suffered from hyperthyroidism, systemic lupus erythematosus, membranoproliferative glomerulonephritis and cryoglobulinemia, respectively. All the 6 stage I patients had achieved complete remission (CR) without relapses 17-84 months (median 44 months) after treatment. Excluding a stage IV patient with follicular lymphoma who died at 3.5 months without treatment, CR was achieved in all of the remaining 16 patients. However, a high relapse rate (9/16, 56%) was noted in stage II-IV patients. These patients tended to relapse in the original sites, but achieved CR again after chemotherapy or radiotherapy. One patient with MALT lymphoma developed histologic transformation into diffuse large lymphoma during relapse and died of refractory disease. Overall, only 2 patients succumbed. The overall survival and relapse-free survival rates at 5 years were 94.7 and 51.4%, respectively. Thus, salivary gland lymphoma proved to be an indolent disease., (2004 S. Karger AG, Basel.)

Background: Androgen receptors are located throughout the brain, especially in regions involved with learning and memory. Different lengths of a CAG (glutamine) repeat polymorphism in exon 1 of the androgen receptor gene may influence androgen action, with longer repeat lengths conferring decreased androgen sensitivity., Methods: We sought to determine if this CAG polymorphism was associated with cognition in older men., Results: Among 301 community-dwelling white men (mean age, 73.0 +/- 7.1), greater CAG repeat length was associated with lower scores on three cognitive tests (p <.05 for all). In addition, 12 participants (9.8%) had cognitive impairment in the low tertile of CAG repeat length whereas 29 (16.3%) had cognitive impairment in the two higher tertiles (odds ratio = 1.8; 95% confidence interval =.9-3.7)., Conclusions: Research should be directed at identifying the mechanism for this association and to determine if treatment with testosterone prevents cognitive decline.

Unlabelled: In a large cohort of U.S. women aged 65 and older, we report the relationships of BMD measured at several sites, and subsequent fracture risk at multiple sites over > 8 years of follow-up. Although we found almost all fracture types to be related to low BMD, the overall proportion of fractures attributable to low BMD is modest., Introduction: Although several studies have reported the relationship between bone mineral density (BMD) and subsequent fracture risk, most have been limited by short follow-up time, BMD measures at only one or two sites, or availability of data for only select fracture types., Materials and Methods: In the multicenter Study of Osteoporotic Fractures (SOF), we studied the relationship of several different BMD measures to fracture risk of multiple types in 9704 non-black women aged 65 and older. We previously reported on the relationship of peripheral BMD measures to risk of several types of fracture during an average 2.2-year follow-up period. In this expanded analysis, we present results of the relationship of both peripheral and central BMD measures and fractures of multiple types during 10.4 and 8.5 years of follow-up, respectively. We also report population attributable risk (PAR) estimates for osteoporosis and risk of several types of fracture., Results: Our results show that almost all types of fractures have an increased incidence in women with low BMD. However, hip BMD is somewhat more strongly related to most of the fracture types studied than spine or peripheral BMD measures. Nonetheless, the proportion of fractures attributable to osteoporosis (based on a standard definition of osteoporosis) is modest, ranging from < 10% to 44% based on the most commonly used definition of osteoporosis (BMD T-score < -2.5)., Conclusion: Finding effective prevention strategies for fractures in older women will require additional interventions beside preventions for bone loss, such as prevention of falls and other fracture risk factors.

t(X;11)(q24;q23) is a recurring chromosomal translocation in pediatric acute myeloid leukemia. The rearrangement results in fusion of MLL at 11q23 with SEPT6 at Xq24. Here, we report the identification of an MLL-SEPT6 fusion transcript in an infant with acute myeloid leukemia (AML)-M4. Reverse transcription-polymerase chain reaction confirmed the presence of an MLL-SEPT6 fusion transcript composed of exon 8 of MLL and exon 2 of SEPT6, but the absence of the reciprocal SEPT6-MLL fusion transcript. Sequence analysis of the genomic break junctions in MLL and SEPT6 suggested that the rearrangement in this case was the result of an insertion of the inverted Xq24 segment, which contained the 3' region of SEPT6 intron 1 and up to 950 kb centromeric to SEPT6, into MLL intron 8. This is a novel type of chromosomal rearrangement leading to the MLL-SEPT6 fusion. The presence of deletions, duplications, and non-template DNA sequence at the break junctions suggested that the DNA damage-repair machinery is likely to be involved in the translocation events., (Copyright 2003 Wiley-Liss, Inc.)

Background: Chinese herbal medicines are commonly used to improve general health and well-being despite limited scientific data to support their efficacy. We conducted a randomized, double-blind, placebo-controlled trial to determine whether an herbal remedy that is used widely in China was associated with changes in quality of life, energy, memory, sexual function, and qi (the Chinese concept of "vital energy" that is important in general health)., Methods: Residents (n = 237) of Beijing, China, who were aged > or =60 years and had self-reported decreased energy, memory, or sexual function, were randomly assigned to take four tablets of a Chinese herbal formula or of an identical placebo, three times a day for 30 days. Patients returned for one follow-up visit after 30 days for assessment of all outcomes. The main outcome measures were changes in quality of life at 30 days as measured by the 12-Item Short Form Health Survey (SF-12) Mental and Physical Component Summary scales., Results: Use of Chinese herbs was associated with a small benefit in the Mental Component scale (difference of 1.9 points; 95% confidence interval [CI]: 0.1 to 3.6) and no benefit in the Physical Component scale (difference of -0.1 points; 95% CI: -1.7 to 1.5) as compared with placebo. A small improvement in the qi scale was no longer significant after adjusting for baseline differences in this score between groups. There was no improvement in physical performance, memory, or sexual function. The herbal product was well tolerated., Conclusion: Short-term use of a mixture of Chinese herbs was associated with a small benefit in one measure of mental health that is of unclear clinical importance.

Objective: To test the hypothesis that use of lipid-lowering drugs reduces the risk of breast cancer in older women., Methods: This was a multicenter prospective cohort study conducted at four community-based clinical centers in the United States, including 7528 Caucasian women, mean age 77 years. The main outcome measure was incident breast cancer confirmed by medical record and pathology reports identified over an average of 6.8 years (244 cases)., Results: The use of lipid-lowering drugs was reported by 576 women (7.7%). The age-adjusted incidence of breast cancer was 3.1/1000 person-years among statin users, 1.4 among women using other lipid-lowering agents, and 5.0 among nonusers. After adjustment for age and body weight, the relative risk (RR) of breast cancer among statin users was 0.28 (95% confidence intervals [CI] 0.09-0.86), and among women who used other lipid-lowering drugs, it was 0.37 (95% CI 0.14-0.99) in comparison to nonusers. The combined group of lipid-lowering drug users had a 68% reduction in the risk of breast cancer (95% CI 32%-85%). Further adjustment for hormone use, family history of breast cancer, mammography use, or other risk factors did not alter the results., Conclusions: Older women who use lipid-lowering drugs may have a reduced risk of breast cancer. Given the widespread use of statins in older women, these results could have a large public health impact. However, these findings need confirmation in other prospective studies, as they were based on a small number of breast cancer events.

Background: The clinical relevance of mutations of the FMS-like tyrosine kinase 3 (FLT3) gene in specific cytogenetic subgroups is not clear. The authors examined internal tandem duplication (ITD) and Asp835 mutations of FLT3 in patients with acute promyelocytic leukemia (APL) to determine the incidence of these mutations and to analyze the results for correlations with clinicohematologic features and outcome., Methods: Bone marrow samples from 107 patients with APL were analyzed. Isoforms of PML-RAR alpha were identified using a reverse transcription-polymerase chain reaction assay. A standard polymerase chain reaction (PCR) assay was used to detect FLT3/ITD mutations. Asp835 mutations were analyzed by PCR amplification of exon 20 followed by EcoRV digestion. All aberrant PCR products subsequently were sequenced., Results: Twenty-two patients had FLT3/ITD mutations: 9 of 63 patients with L-type PML/RAR alpha, 13 of 34 patients with S-type PML/RAR alpha, and 0 of 10 patients with V-type PML/RAR alpha (P = 0.005). The incidence of FLT3/ITD mutations was significantly higher in patients with S-type PML/RAR alpha than in patients with L-type PML/RAR alpha or V-type PML/RAR alpha. Twenty patients had Asp835 mutations (L-type PML/RAR alpha: n = 11; S-type PML/RAR alpha: n = 8; V-type PML/RAR alpha: n = 1). The frequency of Asp835 mutations was not significantly different among patients with different PML/RAR alpha isoforms (P = 0.582). Three patients had both ITD and Asp835 mutations. The microgranular variant (M3v) form of leukemia was found to be associated with a higher frequency of ITD (P = 0.002) but not with a higher frequency of Asp835 mutations (P = 1.000); analysis of clinicohematologic variables revealed no significant differences in FLT3 mutation incidence among other patient subgroups. There was no significant difference in complete remission rate, overall survival, or event-free survival between patients with ITDs and those without ITDs or between patients with Asp835 mutations and those without Asp835 mutations., Conclusions: The current study found that ITD or Asp835 mutations of the FLT3 gene were present in 36.4% of patients with APL; however, these mutations had no prognostic impact. FLT3/ITD frequently was associated with S-type PML/RAR alpha and with the M3v form of leukemia., (Copyright 2003 American Cancer Society.DOI 10.1002/cncr.11636)

Objective: Depression and depressive symptoms have been associated with increased rates of mortality in a variety of populations, but have not been studied in frail, community-living elderly persons. The authors determined whether depressive symptoms were associated with mortality in a frail, multi-ethnic cohort., Methods: Authors conducted a prospective study of 250 participants enrolled in a program for patients in the community eligible for nursing home placement, determining the occurrence of depressive symptoms at enrollment and survival after 18 months of follow-up, using Cox proportional-hazards models to determine whether depressive symptoms were independently associated with mortality., Results: At enrollment, 73 of the participants (29%) were judged to have depression. Among depressed participants, 26% died, versus 17% of those with fewer depressive symptoms. After they adjusted for various demographic factors, 18-month mortality was almost 70% higher in those with depressive symptoms than in those with fewer symptoms. Women with more depressive symptoms showed nearly a 2.5-fold increase in risk of mortality than women with fewer symptoms, whereas more depressed men were not at increased risk of mortality compared with less depressed men., Conclusion: Depressive symptoms constitute a risk factor for mortality in frail elderly persons.

We report the detection of erythropoietin mRNA in the tumor from a patient with neuroblastoma and erythrocytosis. A 2-year-old girl with neuroblastoma presented with hemoglobin 21.3 g/dL, red blood cells 8.03 x 1012/L, and hematocrit 0.641. The serum erythropoietin level was 26.54 mU/mL. The hemoglobin, hematocrit, and serum erythropoietin level were within normal ranges 3 months after surgical excision of the tumor. RT-PCR analysis showed that erythropoietin mRNA expression in the tumor tissue was as high as that of normal renal tissue. These results conclusively demonstrated that the tumor was the site of the ectopic erythropoietin production.

The Bcr-Abl protein-tyrosine kinase is implicated in the development of chronic myeloid leukemia. The potential role of protein-tyrosine phosphatase in the regulation of Bcr-Abl signaling was explored. First, expression patterns of tyrosine phosphatases in leukemic cell lines were investigated using degenerate primers for reverse transcription-PCR followed by cloning and sequencing of the cDNA. Distinct patterns of distribution of phosphatase were found in erythroid and myeloid leukemic cell lines. Whereas some phosphatases were ubiquitously expressed, others were limited to specific cell types. Surprisingly, a previously cloned "lymphocyte-specific" phosphatase, Lyp, was frequently detected in a number of myeloid cell lines as well as normal granulocytes and monocytes. Lyp was localized to the cytosol, and overexpression of Lyp caused reduction in the phosphorylation levels of multiple proteins in KCL22 chronic myeloid leukemia blast cells including Cbl, Bcr-Abl, Erk1/2, and CrkL. Co-expression of Lyp and Bcr-Abl in Cos-7 cells resulted in decreased levels of Bcr-Abl, Grb2, and Myc. Overexpression of Lyp markedly suppressed anchorage-independent clonal growth of KCL22 cells. Taken together, the data suggest that Lyp may play an antagonistic role in signaling by the Bcr-Abl fusion protein.

Objective: To assess the relationship between social network size and prospective mortality risk among a large sample of older, Caucasian women., Method: The study included 7524 Caucasian community-dwelling women, age 65 or older (mean age = 74.1), who participated from four U.S. communities. Study participants completed a protocol that included anthropomorphic and health assessments at baseline and the Lubben Social Network Scale at year 2. We followed participants for an average of 6 years after they had completed the year-2 assessment. We used hospital records and a copy of the participant's official death certificate to document mortality and cause of death in accordance to ICD-9 revision codes., Results: A total of 1451 deaths (19.3% of sample) were observed over follow-up, 215 (3.4%) due to cardiovascular causes. Higher social network scores were a robust predictor of lower multivariate-adjusted mortality (RR = 0.92, 95% CI = 0.86-0.98), controlling for age, comorbid disease, body mass, smoking, depression, and education. However, social network benefits were attenuated after controlling for marital status. Married participants showed lower total (RR = 0.83, 95% CI = 0.74-0.94) and CVD (RR = 0.59, 95% CI = 0.43-0.81) covariate-adjusted death rates compared with unmarried participants., Conclusions: Social network scores and marriage were each associated with reduced prospective mortality risk among older women. The relationships shown here suggest that much of the protection afforded by larger social networks in older women results from marriage rather than other forms of social relationships. Mechanisms at the physiological or behavioral level explaining social relationship benefits remain important areas for future research.

We have shown that the CBL gene at 11q23.3, telomeric to MLL, was fused to MLL in an adult patient with de novo acute myeloid leukemia (FAB-M1). Southern blot analysis indicated that the MLL rearrangement was involved in the chromosomal abnormality. cDNA panhandle polymerase chain reaction identified the fusion transcript, in which MLL exon 6 was fused in-frame with CBL exon 8. Long-distance PCR amplified the genomic junction region, which involved the fusion of the 3' portion of an Alu element in intron 6 of MLL with the 5' portion of an Alu element in intron 7 of CBL. The absence of extensive sequence similarity at both breakpoints of MLL and CBL indicated that the recombination was not generated through homologous recombination. MLL and CBL are located between STS markers D11S939 and D11S924. Analysis of the sequence demonstrated that the transcriptional orientation of both genes at 11q23.3 is from centromere to telomere. The results of Southern blotting in conjunction with fluorescence in situ hybridization suggest that the MLL-CBL fusion was the result of an interstitial deletion. CBL, a proto-oncogene, functions as a negative regulator of several receptor protein-tyrosine-kinase signaling pathways and as an adaptor protein in tyrosine phosphorylation-dependent signaling. CBL is the second gene at 11q23.3 found to fuse with MLL., (Copyright 2003 Wiley-Liss, Inc.)

Objective: Fewer Chinese subjects in Beijing have hip osteoarthritis (OA) compared with whites in the United States, but as many or more Chinese subjects have knee OA. If these differences are due to a systemic predilection for disease, then the prevalence of hand OA, the best indicator of generalized disease, should be different in China. The goals of this study were to estimate the prevalence of hand OA among elderly Chinese in Beijing, and to compare it with that among elderly whites in the United States., Methods: We recruited a random sample of Beijing residents ages > or =60 years. Subjects answered questions on joint symptoms and provided posteroanterior radiographs of the hand. The protocol was identical to that used in the Framingham, Massachusetts OA Study. The hand radiographs from the Beijing OA Study were read intermingled with films from the Framingham OA Study. We defined a hand joint as having radiographic OA if it had a Kellgren and Lawrence grade >/=2. Symptomatic OA was present when both radiographic OA and self-reported pain were present in the same joint. We classified a subject as having radiographic or symptomatic hand OA if at least 1 hand joint had radiographic or symptomatic OA. We estimated the prevalence of hand OA in elderly subjects in Beijing and compared it with the prevalence of hand OA in elderly subjects from Framingham, using an age-standardized prevalence ratio., Results: We obtained both symptom information and hand radiographs from 2,525 subjects. Despite the older age of the group, only 44.5% of men and 47.0% of women had radiographic hand OA. Symptomatic hand OA occurred in 3.0% of men and 5.8% of women. Compared with white men in Framingham, the Beijing Chinese men had a much lower prevalence of radiographic hand OA (age-adjusted prevalence ratio 0.64, 95% confidence interval [95% CI] 0.58-0.70) and symptomatic hand OA (age-adjusted prevalence ratio 0.25, 95% CI 0.16-0.34). The proportion of multiple hand joints affected by OA in Chinese men was also significantly lower than in white men. A similar magnitude of difference was also observed in the women. However, prevalence of symptomatic OA at the metacarpophalangeal (MCP) joints in Chinese men and prevalence of radiographic OA at the MCP joints in Chinese women were similar to those in their white counterparts in Framingham., Conclusion: Elderly Chinese subjects in Beijing had a much lower prevalence of hand OA than did elderly whites in Framingham, Massachusetts. Coupled with the exceedingly low prevalence of hip OA in China, these results may suggest that the overall predilection for OA is less among subjects in China than among whites in the United States.

Senescence-acceleration-prone mice (SAMP8) provide a model to study the influence of early postnatal sound exposure upon the aging auditory midbrain. SAMP8 were exposed to a 9-kHz monotone of either 53- or 65-dB sound pressure level during the first 30 postnatal days, the neurons in the auditory midbrain responding selectively to 9 kHz were localized by c-fos immunohistochemistry and the following parameters were compared to control SAMP8 not exposed to sound: mortality after sound exposure, dendritic spine density, and quantitative neurochemical alterations in this 9-kHz isofrequency lamina. For morphometric analysis, animals were examined at 1, 4, and 8 months of age. Serial sections of the inferior colliculus were Golgi impregnated or stained immunohistochemically for the expression of epsilon1 subunit of NMDA receptor or GABA. Mortality after exposure to 53 dB was the same as in controls, but was markedly increased from 7 months of age onward after postnatal exposure to 65 dB. No gross morphological alterations were observed in the auditory midbrain after sound exposure. However, sound exposure to 53 or 65 dB significantly reduced dendritic spine density by 11% at 4 months or by 11-17% both at 1 and 4 months of age, respectively. The effect of sound exposure upon neurons expressing the NMDAepsilon1 subunit was dose-dependent. Increasing with age until 4 months in control mice and remaining essentially stable thereafter, the percentage of NMDAepsilon1-immunoreactive neurons was significantly elevated by 40-66% in 1- and 8-month-old SAMP8 exposed to 53 dB, whereas no significant effect of 65 dB was apparent. The proportion of GABAergic cells declined with age in controls. It was significantly decreased at 1 month after 53 and 65 dB sound exposure. In contrast, it was elevated at later stages, being significantly increased at 4 months after exposure to 53 dB and at 8 months after exposure to 65 dB. The total cell number in the 9-kHz isofrequency lamina of SAMP8 decreased with age, but was not affected by exposure to either 53 or 65 dB. The present results indicate that early postnatal exposure to a monotone of mild intensity has long-term effects upon the aging auditory brain stem. Some of the changes induced by sound exposure, e.g., decline in spine density, are interpreted as accelerations of the normal aging process, whereas other effects, e.g., increased NMDAepsilon1 expression after 53 dB and elevated GABA expression after both 53 and 65 dB, are not merely explicable by accelerated aging.

Objectives: To characterize the functional trajectories during the last 2 years of life of patients with progressive frailty, with and without cognitive impairment, and to assess whether it was possible to identify discrete functional indicators that signal the end of life., Design: A retrospective analysis of functional trajectories during the last 24 months of life., Setting: Twelve demonstration sites of the Program of All-inclusive Care for the Elderly (PACE). PACE cares for frail older people who meet criteria for nursing home placement, with the goal of keeping the patient at home., Participants: Nine hundred seventeen patients who died while enrolled in PACE., Measures: At PACE entry and every 3 months thereafter, data were collected about the degree of dependence (none, partial, or full) in bathing, eating, and walking and the degree of incontinence (none, bladder, or bowel). Cognitive impairment was defined as six or more errors on the Short Portable Mental Status Questionnaire. To describe the end-of-life trajectories of patients, data were analyzed from observational windows of time, beginning with the patients' dates of death and extending backward in time to 24 months before death. Each analytical window was 3 months in duration. For each of the functional measures, the probability of functional deterioration in the last 2 years of life in patients with (64%) and without (36%) cognitive impairment was also compared., Results: The mean age at death was 84; 69% of patients were women. For patients with and without cognitive impairment, a prolonged, steady increase in the rates of functional dependence that were evident at least 1 year before death, rather than sudden increases in functional dependence shortly before death, characterized the functional trajectories. It was not possible for any of the four measures to detect a time point before death at which there was an abrupt decline in function likely to signal impending death. For each measure, patients with cognitive impairment declined earlier, were more likely than patients without cognitive impairment to have the maximal level of dependence in the 0- to 3-month window before death (e.g., 56% vs 30% for mobility, P <.001), and were more likely to decline in the 2 years before death (e.g., 56% vs 36% for mobility, P <.001)., Conclusion: Patients with advanced frailty, with or without cognitive impairment, have an end-of-life functional course marked by slowly progressive functional deterioration, with only a slight acceleration in the trajectory of functional loss as death approaches. Patients with cognitive impairment have particularly high rates of functional impairment at the time of death. These results suggest that end-of-life care systems that are targeted toward patients with functional trajectories clearly suggesting impending death (such as the Medicare hospice benefit) are poorly suited to older people dying with progressive frailty.

Objectives: To examine the relationship between race and mortality in frail community-dwelling older people with access to a program providing comprehensive access and coordination of services., Design: A longitudinal cohort study., Setting: Twelve nationwide demonstration sites of the Program of All-Inclusive Care for the Elderly (PACE) from 1990 to 1996. PACE provides comprehensive medical and long-term care services for nursing home-eligible older people who live in the community., Participants: Two thousand two white patients and 859 black patients., Measurements: Patients were followed after enrollment until death or the end of the follow-up period. Time from enrollment to death was measured with adjustment of the Cox proportional hazards model for comorbid conditions, functional status, site, and other demographic characteristics., Results: Black patients were younger than white patients (mean age 77 vs 80, P <.001) but had worse functional status (mean activity of daily living (ADL) score 6.5 vs 7.2, P <.001) on enrollment. Survival for black and white patients was 88% and 86% at 1 year, 67% and 61% at 3 years, and 51% and 42% at 5 years, respectively (unadjusted hazard ratio (HR) for black patients = 0.77; 95% confidence interval (CI) = 0.67-0.89). After adjustment for baseline comorbid conditions, functional status, site, and demographic characteristics, black patients still had a lower mortality rate (HR = 0.77; 95% CI =.65-0.93). The survival advantage for black patients did not emerge until about 1 year after PACE enrollment (HR for first year after enrollment = 0.97; 95% CI = 0.72-1.31; HR after first year = 0.67; 95% CI = 0.54-0.85, P-value for time interaction <.001). During the first year of enrollment, black patients were more likely to improve and less likely to decline in ADL function than white patients (P <.001)., Conclusion: In PACE, a system providing access to and coordination of comprehensive medical and long-term care services for frail older people, black patients have a lower mortality rate than white patients. This survival advantage, which emerges approximately 1 year after PACE enrollment, may be related to the comprehensive access and coordination of services provided by the PACE program.

The reactivities of benzynes and metal-carbyne complexes are normally associated with the triple bond units. However, we have now found that electrophiles do not attack the formal osmium-carbon triple bond of osmabenzyne complex 1. Instead, 1 undergoes electrophilic substitution reactions-the typical reactions of aromatic systems.

Objectives: To determine whether the rate of bone loss predicts subsequent cognitive decline independently of baseline bone mass and whether apolipoprotein E (ApoE) genotype explains the association., Design: A prospective cohort study., Setting: Clinical centers in Baltimore, Maryland; Minneapolis, Minnesota; Pittsburgh, Pennsylvania; and Portland, Oregon., Participants: Four thousand four hundred sixty-two women aged 70 and older (mean = 75.8) participating in the Study of Osteoporotic Fractures., Measurements: Total hipbone mineral density (BMD) was measured 2 and 6 years after enrollment (mean follow-up = 3.5 years), and expressed as annualized percentage rate of bone change. A modified Mini-Mental State Examination (mMMSE) was administered at 6 and 10 years (mean follow-up = 4.5 years) and defined cognitive decline as a decline of three or more points on repeat mMMSE score. ApoE genotype information was available on 883 women., Results: Cognitive decline occurred in 12% of the women with the least bone loss (by quartile), 14% in the second, 16% in the third, and 20% in those with the greatest bone loss. After adjustment for age, education, stroke, functional status, estrogen use, body mass index, and smoking, the results were similar. Those who lost the most BMD were almost 40% more likely than women in the lowest quartile to develop cognitive decline in the multivariate model (odds ratio (OR) = 1.4, 95% confidence interval (CI) = 1.1-1.8). A similar association between hipbone loss and cognitive decline was observed in the multivariate model further adjusting for ApoE e4 (OR = 1.5, 95% CI = 0.8-2.7)., Conclusions: Women with more rapid hipbone loss were more likely to develop cognitive decline than those who had lower rate of loss (or who gained bone mass). Differences in functional status, estrogen use, and ApoE did not explain this association. Further investigation is needed to determine the mechanisms that link osteoporosis and cognitive decline.

Objectives: To determine the rate of hip fracture and risk factors associated with hip fractures in disabled older persons who enroll in the Program of All-Inclusive Care for the Elderly (PACE), a program providing comprehensive care to community-living nursing-home-eligible persons., Design: Prospective cohort study between January 1990 and December 1997., Setting: The twelve PACE demonstration sites: San Francisco, California; Columbia, South Carolina; Detroit, Michigan; Denver, Colorado; East Boston, Massachusetts; El Paso, Texas; Milwaukee, Wisconsin; Oakland, California; Portland, Oregon; Rochester, New York; Sacramento, California; and the Bronx, New York., Participants: Five thousand one hundred eighty-seven individuals in PACE; mean age 79, 71% female, 49% white, 47% with dementia., Measurements: Functional status, cognitive status, demographics, and comorbid conditions were recorded on all the participants, who were tracked for occurrence of a hip fracture. The goals were to determine the rate of hip fracture and identify risk factors., Results: Two hundred thirty-eight hip fractures (4.6%) occurred during follow-up. The rate of hip fracture was 2.2% per person-year. Four independent predictors of hip fracture were identified using Cox proportional hazard analysis: age of 75 and older (adjusted hazard ratio (HR) = 2.0, 95% confidence interval (CI) = 1.4-2.8); white ethnicity (HR = 2.1, 95% CI = 1.6-2.8); ability to transfer independently to and from bed, chair, and toilet (HR = 3.0, 95% CI = 1.2-7.2); and five or more Short Portable Mental Status Questionnaire errors (HR = 1.6, 95% CI = 1.3-2.1). The incidence of hip fracture ranged from 0.5% per person-year in persons with zero to one independent risk factors to 4.7% per person-year in those with all four independent risk factors., Conclusions: The rate of hip fracture in this cohort of disabled community-living older adults was similar to that reported in nursing home cohorts. Older age, white race, ability to transfer independently, and cognitive impairment were independent predictors of hip fracture. Persons with these risk factors should be targeted for preventive interventions, which should include strategies for making transferring safer.

We describe a rare presentation of extra-medullary relapse in an adolescent boy with Philadelphia chromosome-positive acute lymphoblastic leukemia who had undergone allogeneic bone marrow transplantation after first remission. In spite of enduring bone marrow remission, the patient experienced a local relapse in the left proximal femur within 3 years of the transplant. The findings from radiography, bone scintigraphy, and chimerism analysis with short tandem repeats as well as bone marrow aspirates taken via the iliac crests were indeterminate. Magnetic resonance imaging at the onset of hip pain was characterized by decreased signal intensity of the left proximal femur, a finding characteristic of bone marrow edema. Confirmation of extra-medullary relapse of the proximal femur was delayed until histologic proof of the computed tomography-guided biopsy samples was obtained. Overt bone marrow relapse was identified 14 months later. Reestablishment of normal donor hematopoiesis was achieved with reinduction chemotherapy.

Analysis of internal tandem duplications of FLT3 (FLT3/ITD) was performed on bone marrow samples obtained at diagnosis and relapse from 108 adult patients with de novo acute myeloid leukemia (AML) to determine the role of this mutation in leukemic relapse. Eighty-three patients had wild-type FLT3 at both diagnosis and relapse, 16 had FLT3/ITD at both stages, whereas 8 had acquired the mutation and 1 had lost it at relapse. Using Genescan analysis, we found that FLT3/ITD levels at first relapse were significantly higher than those at diagnosis (mean +/- SE, 40.5% +/- 4.8% versus 17.9% +/- 3.6%, P <.001). The increase in mutation levels at relapse as compared with diagnosis did not correlate with the difference in blast cell percentages at both stages (P =.777). A hemizygous deletion of wild-type FLT3 was found in 4 patients at relapse compared to none at diagnosis. Nine of the 11 patients carrying a single mutation at diagnosis relapsed with an identical mutation. All 6 patients with more than one FLT3/ITD mutation at diagnosis showed changes in mutation patterns and levels at first relapse; however, each patient retained at least one mutation in the relapse sample. The changes of mutation patterns had implications for the monitoring of minimal residual disease. Our results suggest that FLT3/ITD may contribute as the initial transforming event in AML, and relapse can reflect the selection and outgrowth of a mutant clone or evolution of a new clone harboring this mutation.

The authors sought to determine whether recombinant human thrombopoietin (TPO) acts synergistically with other cytokine(s) on burst-forming unit-erythroid (BFU-E)-derived and colony-forming unit-granulocyte/macrophage (CFU-GM)-derived colony formations from cord blood of premature neonates. Cord blood nonadherent mononuclear cells (MNC) from normal premature neonates were cultured in a methylcellulose system. When cultured with 1 x 10(4) MNC/mL, erythropoietin (EPO) 2 U/mL, interleukin-3 (IL-3) 50 ng/mL, and/or TPO 100 ng/mL, the addition of TPO to EPO gave rise to more BFU-E-derived colonies (p = .000). The addition of TPO to EPO + IL-3 gave rise to more BFU-E-derived colonies (p = .002) also. TPO synergizes erythropoiesis from cord blood of premature neonates. Likewise, the addition of TPO to G-CSF gave rise to more CFU-GM-derived colonies (p = .000) also. TPO synergizes myelopoiesis from cord blood of premature neonates. Thus, TPO has synergistic effects on both erythropoiesis and myelopoiesis from cord blood of premature neonates.

Essential thrombocythemia (ET) is a heterogeneous disorder in which the clonality of hematopoiesis varies. The clinical significance of clonality status in ET remains to be determined. We used the human androgen receptor gene (HUMARA)-polymerase chain reaction assay to investigate X-chromosome inactivation patterns (XCIPs) and their value in predicting vascular complications in 89 female patients with ET. Fifty-four (68.4%) patients had a clonal pattern of XCIP, and 15 (19.0%) had a polyclonal pattern. The remaining 20 patients had either an ambiguous or a homozygous pattern of XCIP and were therefore excluded from further analysis. Patients with clonal XCIPs were older (P =.029) and were at greater risk for thrombosis (P =.007) than were those with polyclonal XCIPs. We did not find a correlation between the occurrence of hemorrhage and XCIP (P =.492). Advanced age was predictive of thrombosis and hemorrhage. Platelet count did not influence the risk for vascular complications. Hypertension was significantly correlated with thrombotic events (P =.002), whereas diabetes mellitus and hypercholesterolemia were of no predictive value. In a multivariate analysis, age was the significant predictor of thrombosis (P =.030); however, XCIPs (P =.083) and hypertension (P =.073) tended to predict thrombosis. Our results suggest that older patients who have clonal XCIPs or hypertension are at increased risk for thrombosis and should be monitored closely for this complication.

Objective: To compare the prevalence of osteoarthritis (OA) of the hip among elderly persons in China and the US., Methods: We recruited a population-based sample of 1,506 persons (82% of those enumerated) ages > or = 60 years living in Beijing, China. Subjects answered questions about joint symptoms and underwent radiography of the pelvis. Radiographs of the Beijing subjects were intermingled with hip radiographs of white women ages > or = 65 years from the Study of Osteoporotic Fractures (SOF) and white men and women ages 60-74 years from the First National Health and Nutrition Examination Survey (NHANES-I) and were then interpreted. Radiographic hip OA was defined as the presence of 1 of the following 3 findings in either hip: minimum joint space of < or = 1.5 mm, definite osteophytes and joint space narrowing, or > or = 3 radiographic features of OA. Symptomatic hip OA was defined as both radiographic OA and hip pain., Results: The crude prevalence of radiographic hip OA in Chinese ages 60-89 years was 0.9% in women and 1.1% in men; it did not increase with age. Chinese women had a lower age-standardized prevalence of radiographic hip OA compared with white women in the SOF (age-standardized prevalence ratio 0.07) and the NHANES-I (prevalence ratio 0.22). Chinese men had a lower prevalence of radiographic hip OA compared with white men of the same age in the NHANES-I (prevalence ratio 0.19). There were no cases of symptomatic hip OA in the Chinese men and only 1 case in the Chinese women; 35 cases were expected in both sexes., Conclusion: This is the first population-based study of hip OA in China to use standardized radiographic methods and definitions. We found that hip OA was 80-90% less frequent than in white persons in the US. Identification of the genetic and environmental factors that underlie these differences may help elucidate the etiology and prevention of hip OA.

Background: Acute illness may lead to long-term losses in older adults' ability to independently perform activities of daily living (ADLs). The magnitude of these losses may differ for patients with and without cognitive impairment. These relationships have not been described for the frailest of older adults whose high rates of acute illness and cognitive impairment put them at the greatest risk for loss of ADL functioning., Methods: We conducted a prospective study of 2593 patients enrolled in a nationwide medical and psychosocial program for frail, community-living, nursing home-eligible patients. We determined the independent and interactive effects of baseline cognitive impairment and admission for an acute illness on change in ADL functioning over 1 year., Results: ADL decline over 1 year occurred in 53% of cognitively impaired patients who were admitted for an acute illness, 38% of cognitively impaired patients who were not admitted for an acute illness, 42% of noncognitively impaired patients who were admitted for an acute illness, and 25% of noncognitively impaired patients who were not admitted for an acute illness (p <.001). The amount of additional decline in ADLs associated with an admission for an acute illness was similar between patients with and without cognitive impairment (-.85 vs -.74; p for interaction =.86). Among patients who were admitted for an acute illness, significant decline in ADL functioning occurred only in the quarter surrounding the acute illness with no evidence of recovery in the months after the acute illness episode., Conclusions: Among frail older adults, loss of ADL functioning over 1 year is independently associated with both acute admission for an acute illness and cognitive impairment. Frail elders, especially those with cognitive impairment, are in need of interventions that reduce the long-term functional consequences of acute illness.

Background: The classification of B-lineage acute lymphoblastic leukemia (ALL) by specific chromosomal translocations may have prognostic implications. Reverse transcriptase-polymerase chain reaction (RT-PCR) assay is a useful tool for the detection of fusion transcript resulting from specific chromosomal translocation of the leukemic cells. In general, fusion transcripts are determined individually, a process which is labor intensive in order to detect all major fusion transcripts., Procedure: We use a multiplex RT-PCR assay to detect both the CML- and ALL-type BCR-ABL transcripts of the t(9;22), all described variants of the E2A-PBX1 transcripts of t(1;19), the MLL-AF4 transcripts of t(4;11), and all described variants of TEL-AML1 (also termed ETV6-CBFA2) of the cryptic t(12;21) in 165 leukemic samples at diagnosis., Results: The study yielded a completely concordant result with those obtained by the individual RT-PCR assay. In this cohort of Taiwan children, the relative frequencies of the four translocations of B-lineage ALL were as following: 6% with ALL-type t(9;22)/BCR-ABL, 7% t(1;19)/E2A-PBX1, 3% t(4;11)/MLL-AF4, and 18% t(12;21)/TEL-AML1, comparable to those in the Western countries., Conclusion: Multiplex RT-PCR assay is an efficient, sensitive, accurate, and cost-effective diagnostic tool, which will likely improve our ability in accurately and rapidly risk-stratifying children with ALL., (Copyright 2002 Wiley-Liss, Inc.)

Background: Recently, an internal tandem duplication of the FLT3 gene (FLT3/ITD) was found in approximately 20% of adult acute myeloid leukemia (AML) cases and associated with a poor outcome. However, there are few studies on FLT3/ITD in childhood AML, and the clinical significance of FLT3/ITD is thus unclear., Methods: FLT3/ITD was analyzed in 80 children with de novo AML. The genomic DNA polymerase chain reaction (PCR) assay was performed to identify FLT3/ITD. Genescan analysis to determine the allelic distribution was then performed for those PCR products with aberrant bands. Direct sequencing of PCR products was also carried out in each sample with FLT3/ITD., Results: The incidence of FLT3/ITD was 11.3% (9 out of 80 patients) in AML, with 25% (3 out of 12 patients) in acute promyelocytic leukemia (APL) and 8.8% (6 out of 68 patients) in non-M3 AML. The size of duplicated fragments varied from 21 base pairs (bp) to 75 bp, and the mutant to wild type ratio of FLT3 ranged from 0.28 to 16.60 in the nine patients with FLT3/ITD. The incidence of FLT3/ITD in childhood AML in patients > 10 years of age was 24%, compared to 5% of those patients

Background: Several genes associated with sporadic Alzheimer's disease have been identified; however, approximately 50% of genetic factors remain unidentified. We investigated whether estrogen receptor 1 (ESR1) polymorphisms are associated with risk of developing cognitive impairment in older women., Methods: A total of 2625 women > or = 65 years of age completed a modified Mini-Mental Status Exam (mMMSE) at baseline and at 6-8 years of follow-up. We defined cognitive impairment as a mMMSE decline of > or = 3 points, follow-up score < or = 20, or a history of physician-diagnosed dementia. The ESR1 polymorphisms, PvuII (P or p) and XbaI (X or x), were coded so that the capital letter signifies the absence of the restriction site., Results: Women with a p allele had a greater age, education, and baseline-score adjusted decline in mMMSE (for PP, Pp, and pp, respectively:.6 +/-.1,.8 +/-.1, and.9 +/-.1 points, p for trend =.01); women with at least one x allele also had greater score decline (XX, Xx, and xx:.7 +/-.1,.7 +/-.1, and.9 +/-.1 points, p for trend =.02). Six percent (n = 166) of the women developed cognitive impairment. Compared to those who did not develop impairment, more women who developed cognitive impairment had a p allele (62% vs. 56%, p =.03; adjusted odds ration (OR) = 1.33; 95% confidence interval [CI], 1.03-1.72) or an x allele (70% vs. 64%, p =.03; adjusted OR = 1.38; 95% CI, 1.06-1.81). There was no interaction with current estrogen use, or with serum estradiol level and ESR1 polymorphisms (p >.10)., Conclusions: Estrogen receptor 1 polymorphisms are associated with risk of developing cognitive impairment. More research is needed to determine the mechanism whereby ESR1 polymorphisms or linked genes influence cognitive function in older women.

Objectives: Recent studies have suggested that estrogen may improve cognitive function or prevent cognitive decline in older women. Little research has been conducted on exogenous or endogenous sex hormones and cognition in older men, yet it has been hypothesized that testosterone, either directly or by conversion to estrogens, may improve cognitive function. We investigated whether serum level of testosterone and estradiol is associated with cognition in older community-dwelling men., Design: A cross-sectional study., Setting: Population-based listings in the Monongahela Valley near Pittsburgh, Pennsylvania., Participants: Three hundred ten men (mean age +/- standard deviation = 73.0 +/- 7.1) who were part of a cohort study., Measurements: We measured cognitive function using the Mini-Mental State Examination (MMSE), Trails B, and Digit Symbol. Sex hormone levels were determined by radioimmunoassay from serum obtained at the time of cognitive testing and analyzed by tertile., Results: No consistent association between total testosterone level and cognitive test scores was observed. However, men with high bioavailable (loosely protein-bound) testosterone had better cognitive test scores on all three tests (P < or =.001). Total estradiol levels were associated with worse cognitive scores on Digit Symbol (P <.001) and Trails B (P =.002), but bioavailable estradiol levels were not associated with cognitive function. Level of sex hormone binding globulin (SHBG) was negatively associated with cognitive scores on all three tests (P < or =.001). After adjusting for age and education, the statistical significance lessened for bioavailable testosterone (MMSE, P =.086; Digit Symbol, P =.047; Trails B, P =.076) and became nonsignificant for SHBG (all cognitive tests P>.10)., Conclusions: Our findings support the hypothesis that higher levels of bioavailable testosterone, but not of bioavailable estradiol, are associated with better cognitive function in older men. In addition, bioavailable measures of testosterone may better reflect hormone levels available to the brain and thus be more closely associated with central nervous system outcomes such as cognition. Future studies, especially randomized trials, should be undertaken to determine whether testosterone may protect against cognitive decline in older men.

The pineal neurohormone melatonin modulates a variety of physiological processes through different receptors. It has recently been reported that the cloned melatonin receptors (MT1, MT2 and Mel1c) exhibit differential abilities to stimulate phospholipase C (PLC) via G(16). Here we examined the molecular basis of such differences in melatonin receptor signaling. Coexpression of MT1 or MT2 with the alpha subunit of G(16) (Galpha(16) ) allowed COS-7 cells to accumulate inositol phosphates in response to 2-iodomelatonin. In contrast, Mel1c did not activate Galpha(16) even though its expression was demonstrated by radioligand binding and agonist-induced inhibition of adenylyl cyclase. As Mel1c possesses an exceptionally large C-terminal tail, we further asked if this structural feature prevented productive coupling to Galpha(16). Eleven chimeric melatonin or mutant receptors were constructed by swapping all or part of the C-terminal tail between MT1, MT2 and Mel1c. All chimeras were fully capable of binding 2-[(125) I]iodomelatonin and inhibiting adenylyl cyclase. Chimeras containing the full-length Mel1c tail were incapable of activating Galpha(16), while those that contained the complete C-terminal region of either MT1 or MT2 stimulated PLC. Incorporation of the extra portion of the C-terminal tail of Mel1c to either MT1 or MT2 completely abolished the chimeras' ability to stimulate PLC via Galpha(16). In contrast, truncation of the C-terminal tail of Mel1c allowed interaction with Galpha(16). Our results suggest that Galpha(16) can discern structural differences amid the three melatonin receptors and provide evidence for functional distinction of Mel1c from MT1 and MT2 receptors.

The Prp19p-associated complex is essential for the yeast pre-mRNA splicing reaction. The complex consists of at least eight protein components, but is not tightly associated with spliceosomal snRNAs. By a combination of genetic and biochemical methods we previously identified four components of this complex, Ntc25p, Ntc85p, Ntc30p and Ntc20p, all of them being novel splicing factors. We have now identified three other components of the complex, Ntc90p, Ntc77p and Ntc31p. These three proteins were also associated with the spliceosome during the splicing reaction in the same manner as Prp19p, concurrently with or immediately after dissociation of U4 snRNA. Two-hybrid analysis revealed that none of these proteins interacted with Prp19p or Ntc25p, but all interacted with Ntc85p. An interaction network between the identified components of the Prp19p-associated complex is demonstrated. Biochemical analysis revealed that Ntc90p, Ntc31p, Ntc30p and Ntc20p form a subcomplex, which, through interacting with Ntc85p and Ntc77p, can associate with Prp19p and Ntc25p to form the Prp19p-associated complex. Genetic analysis suggests that Ntc31p, Ntc30p and Ntc20p may play roles in modulating the function of Ntc90p.

Gastric adenocarcinoma developing concomitantly with a gastric lymphoma of the mucosa-associated lymphoid tissue (MALT) type is rare. Herein, we report a case with a synchronous primary gastric MALT lymphoma and an early adenocarcinoma of the stomach. A 72-year-old patient with initial presentation of weight loss was found with endoscopy to have a large tumor mass in the gastric body. Pathologic examination of biopsies revealed a low-grade MALT lymphoma for which chemotherapy with cyclophosphamide, vincristine, and prednisolone was administered. A gastric adenocarcinoma was found at a different site in the of stomach 3 months after cessation of chemotherapy when there was still residual MALT lymphoma in the stomach. The presence of double neoplasms was established preoperatively. The patient underwent a proximal gastrectomy. Infection with Helicobacter pylori (H. pylori) was detected once in the repeated endoscopic gastric biopsies. The occurrence of both gastric MALT lymphoma and gastric adenocarcinoma was reviewed and the association of H. pylori infection with both malignancies is discussed.

The family of melatonin receptors is composed of the mt1, MT2, and Mel1c subtypes. The Mel1c is further divided into one long and two short isoforms. A recent study has shown that, unlike mt1 and MT2, the long form of Mel1c is incapable of activating the pertussis toxin-insensitive G16. Here we used three well-characterized Galphaq chimeras to explore the coupling specificity of the melatonin receptors. The qi5, qo5, and qz5 chimeras can link numerous Gi-coupled receptors to the stimulation of phosphoinositide-specific phospholipase C. Both mt1 and MT2 receptors interacted productively with the Galphaq chimeras, while the long form of Mel1c was totally ineffective. Among the Galphaq chimeras, qo5 was less efficiently coupled to the melatonin receptors. Such differential coupling is best explained by structural differences between the melatonin receptors as well as among the Galphaq chimeras. Since the long form of Mel1c receptor possesses an exceptionally large C-terminal tail, we tested the ability of four melatonin receptor C-terminal tail chimeras (Chi 1-4) to interact with the Galphaq chimeras. The presence of the large C-terminal tail of Mel1c in Chi 1 and Chi 3 markedly hindered their coupling to the Galphaq chimeras. On the other hand, the attachment of either the mtl or MT2 C-terminal tail to a Mel1c backbone produced chimeras (Chi 2 and Chi 4) that were capable of activating the Galphaq chimeras. These findings suggest the involvement of C-terminal regions of melatonin receptors in the recognition of G proteins.

1. The bovine Galpha(14) is a member of the G(q) subfamily of G proteins that can regulate phospholipase Cbeta isoforms but the extent to which Galpha(14) recognizes different receptor classes is not known. 2. Galpha(14) was cotransfected with a variety of receptors in COS-7 cells, and agonist-induced stimulation of phospholipase C was then measured. 3. Activation of the type 2 but not type 1 somatostatin receptor in cells coexpressing Galpha(14) stimulated the accumulation of inositol phosphates; functional expression of both subtypes of somatostatin receptors was determined by the ability of somatostatin to inhibit cyclic AMP accumulation. 4. Among the three opioid receptors (mu, delta, and kappa), only the delta receptor was capable of stimulating IP formation when coexpressed with Galpha(14) in COS-7 cells. 5. A panel of G(i)- and G(s)-linked receptors was screened for their ability to stimulate IP accumulation via Galpha(14). The adenosine A(1), complement C5a, dopamine D(1), D(2) and D(5), formyl peptide, luteinizing hormone, secretin, and the three subtypes of melatonin (mt1, MT2, and Xenopus) receptors were all incapable of activating Galpha(14), while the alpha(2)- and beta(2)-adrenoceptors were able to do so. 6. Galpha(14)-mediated stimulation of phospholipase Cbeta was agonist dose-dependent. These data demonstrate that although Galpha(14) can interact with different classes of receptors, it is much less promiscuous than Galpha(15) or Galpha(16).

The human opioid receptor-like (ORL(1)) receptor was tagged with the immunoglobulin G1 Fc fragment at the carboxy-terminus and expressed in human embryonic kidney 293 cells. Expression of the ORL(1)-Fc receptor was confirmed by immunohistochemical staining. The fusion protein was enriched by affinity chromatography and then verified by immunodetection. The function of the ORL(1)-Fc receptor was determined by examining nociceptin/OFQ-induced inhibition of cAMP accumulation. The ORL(1)-Fc receptor inhibited the forskolin-stimulated cAMP accumulation. The inhibitory response was selectively induced by nociceptin/OFQ in a dose-dependent and pertussis toxin-sensitive manner. Our results indicate that the carboxy-terminal Fc-tagged ORL(1) receptor retained the ability to interact with G(i) proteins to inhibit adenylyl cyclase., (Copyright 2000 S. Karger AG, Basel.)

Opioid tolerance and physical dependence in mammals can be rapidly induced by chronic exposure to opioid agonists. Recently, opioid receptors have been shown to interact with the pertussis toxin (PTX)-insensitive Gz (a member of the Gi subfamily), which inhibits adenylyl cyclase and stimulates mitogen-activated protein kinases (MAPKs). Here, we established stable human embryonic kidney 293 cell lines expressing delta-opioid receptors with or without Gz to examine the role of Gz in opioid receptor-regulated signaling systems. Each cell line was acutely or chronically treated with [D-Pen2,D-Pen5]enkephalin (DPDPE), a delta-selective agonist, in the absence or presence of PTX. Subsequently, the activities of adenylyl cyclase, cyclic AMP (cAMP)-dependent response element-binding proteins (CREBs), and MAPKs were measured by determining cAMP accumulation and phosphorylation of CREBs and the extracellular signal-regulated protein kinases (ERKs) 1 and 2. In cells coexpressing Gz, DPDPE inhibited forskolin-stimulated cAMP accumulation in a PTX-insensitive manner, but Gz could not replace Gi to mediate adenylyl cyclase supersensitization upon chronic opioid treatment. DPDPE-induced adenylyl cyclase supersensitization was not associated with an increase in the phosphorylation of CREBs. Both Gi and Gz mediated DPDPE-induced activation of ERK1/2, but these responses were abolished by chronic opioid treatment. Collectively, our results show that although Gz mediated opioid-induced inhibition of adenylyl cyclase and activation of ERK1/2, Gz alone was insufficient to mediate opioid-induced adenylyl cyclase supersensitization.

Adsorbed proteins on biomaterial surfaces determine whether cells adhere, but rheological variables are also critical. Neutrophil adhesion under well-defined radial flow conditions was studied on glass preadsorbed with plasma proteins or plasma protein domain fragments. Fibrinogen, low-molecular-weight kininogen (LK), high-molecular-weight kininogen (HK), cleaved HK (HKa), and recombinant HK domains 3 and 5 (D3 and D5H) were used. The number of adherent cells on the HK and HKa surfaces was less than 10% that found on the fibrinogen absorbed surface. The degree of spreading was minimal and detachment of adherent neutrophils was observed. HK and HKa contain binding sites for both anionic surfaces and neutrophils in the same domain (D5H). When adsorbed to surfaces, HK and HKa did not have the neutrophil binding sites available and therefore exhibited an anti-adhesive effect. Although D5H contains anionic surface binding sites, its small molecular size required a higher number of adsorbed molecules to cover the surface before a significant decrease in cell adhesion was observed. Since LK and D3 do not possess specific anionic surface binding sites, the adsorption of these proteins on glass was very low compared to HK and HKa. Thus, extensive cell adhesion and spreading were observed on the surfaces partially covered with preadsorbed LK and D3.

Three serine-to-alanine mutants of the alpha subunit of the heterotrimeric G protein G(z) (alpha(z)) were examined for their signaling properties in the presence of phorbol ester treatment. All three alpha(z) mutants resembled wild-type alpha(z) in their abilities to inhibit alpha(s)-stimulated type 6 adenylyl cyclase (AC6) and phorbol ester treatment reduced their magnitudes of inhibition. Depending on the permissive condition, the betagamma-mediated stimulation of type 2 adenylyl cyclase (AC2) was differentially regulated by alpha(z) and the three mutants. Mutation of Ser(27) but not Ser(16) of alpha(z) affected the efficient release of betagamma subunits upon receptor activation and abolished the stimulation of phosphorylated but not alpha(s)-stimulated AC2., (Copyright 2000 S. Karger AG, Basel)

A naturally occurring point mutation (R231H) within one of the major 3gamma-binding surface (switch II region) on the a subunit of Gs (alpha(s)) has previously been found to disrupt receptor-mediated activation of Gs. The disruption caused by mutating this conserved residue may be a general phenomenon for all a subunits. Homologous mutants of the alpha subunit of Gz [alpha(z); a negative regulator of adenylyl cyclase (AC)] and G16 (alpha16; a stimulator of phospholipase C) were constructed and examined for receptor-mediated regulation of their corresponding effectors. The mutant alphazR209H cannot be fully activated by the delta-opioid receptor, as indicated by the impairment of the inhibition of alpha(s)-stimulated AC and betagamma-mediated stimulation of AC type II (AC2). Similarly, the mutant alpha16R216H lost the ability to mediate receptor-induced activation of phospholipase C and AC2. The receptor coupling efficacy and promiscuity of alpha16R216H were eradicated. The mutation of the conserved arginine has no observable effect on the constitutive activities of the GTPase-deficient derivatives of both alpha(z) and alpha16. The alpha subunit of Gt1 (transducin; alphat1) attenuated betagamma-mediated stimulation of AC2 by sequestrating free betagamma subunits, but the mutant alphat1R204H showed reduced ability to scavenge betagamma-mediated AC2 activation. Presumably, mutation of the conserved arginine disrupted the subunit interactions in addition to the impairment of receptor interaction.

Interaction of biomaterials with blood components including neutrophils is responsible for some of the clinical complications that have occurred in cardiopulmonary bypass, hemodialysis, and ventricular assist procedures. The possibility of inhibiting the initial adhesion of neutrophils to biomaterials has been studied extensively, but the problem remains unsolved. In this study, we investigated the effect of HK adsorption on polyurethane, a widely used component of extracorporeal and intracorporeal devices. HK and HKa were allowed to adsorb on 4 different charged polyurethanes: noncharged (PU), cationic (NR(4)), anionic (SO(3)), and zwitterionic (GPC) polyurethanes. The effect of kininogen adsorption on neutrophil adhesion, the surface density of the adsorbed kininogen, and the exposure of HK domains 3 and 5 (D(3) and D(5H)), which are responsible for the binding of HK to the neutrophil integrin alpha(m)beta(2) or Mac-1, were examined. On PU, NR(4), and SO(3), kininogen adsorption reached 80% of monolayer coverage when 100 pmol/mL or higher concentration of protein solutions were used. The NR(4) surface adsorbed the most kininogen along with a high exposure of D(3) and D(5H). The availability of D(3) and D(5H) allowed neutrophils to bind to the surface via the Mac-1 receptor; thus, on the NR(4) surface, adsorbed kininogens lost their antiadhesive property, which resulted in a high degree of neutrophil adhesion. Increasing Mac-1 expression by exposure to fMLP increased the neutrophil adhesion on this surface. In contrast, exposure of D(3) and D(5H) on SO(3) was significantly less, because HK binds to anionic surfaces with similar protein sequences used for cell binding. This low binding site exposure preserved the antiadhesive property of HK. GPC was resistant to neutrophil adhesion even in the absence of adsorbed kininogens because of its phosphorylcholine moiety. Thus, both SO(3) coupled with kininogen (or kininogen peptides) and GPC have the potential to markedly reduce neutrophil adhesion to biomaterial devices.

The adhesion of neutrophils on a biomaterial surface depends on the surface chemistry of the material and the cell, as well as the composition and conformation of adsorbed protein and the adherence of other cells when the biomaterial is exposed to circulating blood. In this study, HK and HKa were allowed to adsorb on three different polyurethanes: underivatized (PU-base), quaternized (PU-NR4), sulfonated (PU-SO3). The effect of kininogen adsorption on the degree of neutrophil adhesion was examined. The surface density of the adsorbed protein was also investigated. The PU-NR4 surface adsorbed the most HK and HKa and had the high degree of neutrophil adhesion. Although the surface density of adsorbed HK and HKa on the PU-SO3 surface, the degree of neutrophil on adhesion was significantly lower when compared to the PU-NR4 and PU-base surfaces. HK and HKa contain binding sites for both anionic surfaces and neutrophils in the same domain (D5H). When adsorbed to the anionic PU-SO3 surfaces, HK and HKa did not have the neutrophil binding sites available and therefore, exhibited an anti-adhesive effect. In contrast, the neutrophil binding domains D3 and DsH of adsorbed kininogens were available on the PU-NR4 and PU-base surfaces. Thus, adsorbed kininogens on these two surfaces lost their anti-adhesive property and this led to a high degree of neutrophil adhesion.

In most tissues and cells the opioid receptor-like (ORL1) receptor regulates effectors primarily through the pertussis toxin (PTX)-sensitive guanine nucleotide-binding regulatory proteins (G proteins) Gi/Go. Many Gi-coupled receptors possess additional capability to interact with one or more PTX-insensitive G proteins. Using the betagamma-induced stimulation of type 2 adenylyl cyclase as a readout, we screened the ability of ORL1 receptor to interact with a panel of PTX-insensitive G proteins. In the presence of PTX, activation of the ORL1 receptor resulted in the stimulation of type 2 adenylyl cyclase only in HEK 293 cells coexpressing the alpha subunit of Gz, G12, G14, or G16, but not in cells coexpressing G11, G13, or Gq. Coupling to both Gz and G16 was expected because close relatives of the ORL1 receptor, the opioid receptors, are known to couple productively to these G proteins. ORL1 receptor coupling to either G12 or G14 has not been demonstrated. As predicted by the type 2 adenylyl cyclase assays, activation of the ORL1 receptor resulted in the formation of inositol phosphates in COS-7 cells transiently cotransfected with Galpha14. The ORL1 receptor-mediated stimulation of phospholipase C was found to be Galpha14 dependent, agonist dose dependent, ligand selective, and PTX insensitive. We conclude that G14 can link the ORL1 receptor to regulation of phopholipase C.