Pharmaceutical inhibition of glycogen synthetase kinase-3ß reduces multiple myeloma-induced bone disease in a novel murine plasmacytoma xenograft model.

Multiple myeloma (MM) is a malignancy of plasma cells that accumulate in the bone marrow. MM is incurable with approximately 100 000 patients currently in the United States and 20 000 new cases diagnosed yearly. The malignancy causes displacement of hematopoiesis and formation of osteolytic bone lesions also known as myeloma bone disease (MBD). At diagnosis, 79% of patients suffer from MBD associated with severe pain and increased mortality. Wnt inhibitors secreted by MM cells inhibit osteogenesis and promote osteoclastogenesis, therefore rapid targeting of Wnt inhibitors is necessary to prevent potentially irreversible effects on the stroma, which could lead to incurable MBD. Inhibition of glycogen synthetase kinase-3ß (GSK3ß) causes accelerated Wnt signaling and enhanced osteogenesis in mesenchymal stem/progenitor cells, irrespective of the extracellular concentration of Wnt inhibitors. Our primary goal of this study was to evaluate a GSK3ß inhibitor (6-bromoindirubin-3'-oxime BIO) for amelioration of bone destruction in a murine model of MBD. When measured using histomorphometry, peritumoral BIO administration improved bone quality at the bone-tumor interface and, surprisingly, increased histologically apparent tumor necrosis. Furthermore, in vitro assays demonstrated a proapoptotic effect on numerous MM cell lines. These preliminary data suggest that pharmaceutical GSK3ß inhibition may improve bone quality in myeloma and other malignant bone diseases.

Pandemic influenza A(H1N1)v viruses currently circulating in New Zealand are sensitive to oseltamivir.

New Zealand, like other southern hemisphere countries with a temperate climate, has been in the winter period with seasonal influenza activity. New Zealand has also experienced a dramatic increase in the number of cases of pandemic influenza A(H1N1)v virus. Early reports from the northern hemisphere at the beginning of the pandemic showed that the virus was sensitive to the antiviral drug oseltamivir. In this study we report that pandemic influenza A(H1N1)v viruses currently circulating in New Zealand are sensitive to oseltamivir, but seasonal influenza A(H1N1) viruses - the co-circulating predominant seasonal strain, is resistant to oseltamivir.

Dkk-1-derived synthetic peptides and lithium chloride for the control and recovery of adult stem cells from bone marrow.

It is established that human mesenchymal stem cells (hMSCs) from bone marrow are a source of osteoblast progenitors in vivo and under appropriate conditions, differentiate into osteoblasts ex vivo. Because hMSCs are recovered by iliac crest aspirate and enriched by virtue of their adherence to tissue culture plastic, the cells provide a convenient ex vivo model for the study of osteogenic tissue repair in an experimentally accessible system. Recent advances in the field of skeletal development and osteogenesis have demonstrated that signaling through the canonical wingless (Wnt) pathway is critical for the differentiation of progenitor cell lines into osteoblasts. Inhibition of such signals can predispose MSCs to cell cycle entry and inhibit osteogenesis. Here, we report that synthetic peptides derived from the second cysteine-rich domain of the canonical Wnt inhibitor Dickkopf-1 (Dkk-1) have utility in controlling the growth and recovery of hMSCs from bone marrow stroma. Three peptides corresponding to residues 217-269 in Dkk-1 were each found to enhance the proliferation of hMSCs in culture over 2 days. The most active peptide exhibited agonistic characteristics in that it ablated the proliferation lag observed when cultures of hMSCs receive fresh medium. It also reduced the expression of endogenous Dkk-1 (Gregory, C. A., Singh, H., and Prockop, D. J. (2003) J. Biol. Chem. 278, 28067-28078). When the cytosolic level of beta-catenin was elevated by addition of LiCl to cultures of hMSCs, the peptide also accelerated degradation of beta-catenin on withdrawal of lithium. A second peptide, corresponding to residues 184-204 had preferential and high affinity for hMSCs in the log phase of proliferation. Peptide overlay assays on hMSC lysates confirmed that the peptide bound to a 184-kDa protein corresponding to the molecular mass of LRP6. Cells recovered by this peptide had enhanced osteogenic potential but less chondrogenic potential compared with controls. Because Wnt antagonists increase the number of non-committed hMSCs in culture, they may be of use in increasing the rate of osseous wound healing in vivo by increasing the level of systemically migrating hMSCs. Therefore, such molecules could contribute to the development of a novel family of pharmaceutical agents for the improvement of the healing process in humans.

An Alizarin red-based assay of mineralization by adherent cells in culture: comparison with cetylpyridinium chloride extraction.

Alizarin red S (ARS) staining has been used for decades to evaluate calcium-rich deposits by cells in culture. It is particularly versatile in that the dye can be extracted from the stained monolayer and assayed. This study describes a sensitive method for the recovery and semiquantification of ARS in a stained monolayer by acetic acid extraction and neutralization with ammonium hydroxide followed by colorimetric detection at 405 nm. This method was three times more sensitive than an older method involving cetylpyridinium chloride (CPC) extraction and resulted in a better signal to noise ratio, especially for weakly stained monolayers. The assay facilitates detailed inspection of mineralization by phase microscopy and semiquantification of the entire monolayer by extraction and quantification. The sensitivity of the assay is improved by the extraction of the calcified mineral at low pH and, since the mineral is already stained in a quantitative manner, there is no requirement for an additional colorimetric quantification step. Furthermore, the linear range is much wider than those of conventional assays for calcium, making dilutions of mineral extracts prior to measurement unnecessary. It has a wide range of potential uses including tumor characterization, mesenchymal stem cell evaluation, and osteogenic compound screening. Although more labor intensive than CPC extraction, the protocol is more sensitive and yields more reliable results for weakly mineralizing samples.

Effect of a monetary sanction on immunization rates of recipients of aid to families with dependent children.

CONTEXT: Immunization rates among low-income families have lagged behind those for the general community, with several possible barriers cited in the literature. OBJECTIVE: To evaluate the effect of an initiative aimed at improving immunization rates among low-income preschool children by imposing a sanction on families who failed to provide proof of up-to-date immunization status. DESIGN AND SETTING: Randomized, controlled before-after trial conducted from January 1, 1993, through December 31, 1996, in Muscogee County, Georgia. PARTICIPANTS: A total of 2500 families with children aged 6 years or younger who received Aid to Families with Dependent Children assistance. INTERVENTION: Families in the intervention group (n=1500) were informed that receipt of the welfare benefit for any preschool-aged children was contingent on provision of proof of up-to-date immunization status at the beginning of welfare eligibility and, subsequently, semiannually or annually. Case families in the control group (n=1000) were encouraged to immunize their preschool children but were not informed of any aid sanctions nor did such sanctions apply to them. MAIN OUTCOME MEASURE: Age-appropriate rates of 5 immunizations (measles-mumps-rubella; poliovirus; diphtheria and tetanus toxoids and pertussis; Haemophilus influenzae type b; and hepatitis B), based on examination (with family's written consent) of medical provider records, compared among intervention-group vs control-group families. RESULTS: There were no significant differences at baseline between intervention and control families in immunization rates of preschool children. Families in the intervention group were significantly more likely than families in the control group to have up-to-date immunization status in all 4 years of the study for all 5 immunizations (with 3 exceptions). At age 2 years, 72.4% of children in the intervention group vs 60.6% of those in the control group achieved vaccine series completion, which included 4 diphtheria and tetanus toxoids and pertussis, 3 poliovirus, and 1 measles-mumps-rubella (P<.001). Sanctions were implemented only 11 times. There was relatively little increased burden on the part of families to comply with requirements. CONCLUSION: In our study, a monetary sanction in a population receiving welfare benefits stimulated a significant increase in childhood immunization rates, suggesting that when welfare recipients are given an incentive to keep their children's immunizations up-to-date, most are able to do so. JAMA. 2000;284:53-59

Surveillance for chronic fatigue syndrome--four U.S. cities, September 1989 through August 1993.

PROBLEM/CONDITION: Although chronic fatigue syndrome (CFS) has been recognized as a cause of morbidity in the United States, the etiology of CFS is unknown. In addition, information is incomplete concerning the clinical spectrum and prevalence of CFS in the United States. REPORTING PERIOD COVERED: This report summarizes CFS surveillance data collected in four U.S. cities from September 1989 through August 1993. DESCRIPTION OF SYSTEM: A physician-based surveillance system for CFS was established in four U.S. metropolitan areas: Atlanta, Georgia; Wichita, Kansas; Grand Rapids, Michigan; and Reno, Nevada. The objectives of this surveillance system were to collect descriptive epidemiologic information from patients who had unexplained chronic fatigue, estimate the prevalence and incidence of CFS in defined populations, and describe the clinical course of CFS. Patients aged > or = 18 years who had had unexplained, debilitating fatigue or chronic unwellness for at least 6 months were referred by their physicians to a designated health professional(s) in their area. Those patients who participated in the surveillance system a) were interviewed by the health professional(s); b) completed a self-administered questionnaire that included their demographic information, medical history, and responses to the Beck Depression Inventory, the Diagnostic Interview Schedule, and the Sickness Impact Profile; c) submitted blood and urine samples for laboratory testing; and d) agreed to a review of their medical records. On the basis of this information, patients were assigned to one of four groups: those whose illnesses met the criteria of the 1988 CFS case definition (Group I); those whose fatigue or symptoms did not meet the criteria for CFS (Group II); those who had had an identifiable psychological disorder before onset of fatigue (Group III); and those who had evidence of other medical conditions that could have caused fatigue (Group IV). Patients assigned to Group III were further evaluated to determine the group to which they would have been assigned had psychological illness not been present, the epidemiologic characteristics of the illness and the frequency of symptoms among patients were evaluated, and the prevalence and incidence of CFS were estimated for each of the areas. RESULTS: Of the 648 patients referred to the CFS surveillance system, 565 (87%) agreed to participate. Of these, 130 (23%) were assigned to Group I; 99 (18%), Group II; 235 (42%), Group III; and 101 (18%), Group IV. Of the 130 CFS patients, 125 (96%) were white and 111 (85%) were women. The mean age of CFS patients at the onset of illness was 30 years, and the mean duration of illness at the time of the interview was 6.7 years. Most (96%) CFS patients had completed high school, and 38% had graduated from college. The median annual household income/for CFS patients was $40,000. In the four cities, the age-, sex-, and race-adjusted prevalences of CFS for the 4-year surveillance period ranged from 4.0 to 8.7 per 100,000 population. The age-adjusted 4-year prevalences of CFS among white women ranged from 8.8 to 19.5 per 100,000 population. INTERPRETATION: The results of this surveillance system were similar to those in previously published reports of CFS. Additional studies should be directed toward determining whether the data collected in this surveillance system were subject to selection bias (e.g., education and income levels might have influenced usage of the health-care system, and the populations of these four surveillance sites might not be representative of the U.S. population). ACTIONS TAKEN: In February 1997, CDC began a large-scale, cross-sectional study at one surveillance site (Wichita) to describe more completely the magnitude and epidemiology of unexplained chronic fatigue and CFS.

A blended beam technique to decrease toxic effects of post mastectomy irradiation by combining and sequentially mixing electrons and photons.

We studied 24 patients with Stage II or Stage III breast carcinoma, post total mastectomy, who had received adjuvant loco-regional irradiation (16 patients) or were irradiated for chest wall recurrence (8 patients), along with systemic chemotherapy and/or hormonotherapy. A technique is described for combining and sequentially mixing electron and photon beams. This blended beam method results in less severe acute and chronic skin reactions. Dose distribution to the chest wall and mediastinum are improved compared to the traditional photon-only techniques. At a median followup of four years our expected loco-regional control rates are not compromised. This technique is recommended for selected post mastectomy patients to reduce toxic effects of chest wall irradiation, particularly when chemotherapy lowers skin tolerance and the reserves of the heart, lung and bone marrow.

Epidemiology of chronic fatigue syndrome: the Centers for Disease Control Study.

The US Centers for Disease Control initiated physician-based chronic fatigue syndrome (CFS) surveillance systems in four cities in September 1989 to determine the prevalence, incidence, course and impact of the illness. The participating physicians have referred to our surveillance system 590 patients who were ill during the first two years of surveillance with severe, debilitating, unexplained fatigue for at least the preceding six months. Referred patients were screened for psychiatric disorders preceding, concurrent with, and subsequent to the onset of their fatigue by specially trained nurses using a modified Diagnostic Interview Schedule. Complete health histories were obtained by interview and review of medical records and a basic panel of standard laboratory diagnostic tests were conducted. Four physicians have independently reviewed the health information of 337 of the patients for classification. Approximately 26% of patients referred to the surveillance system met the CFS case definition in all regards, 14% lacked one or more of the required eight symptom criteria, 15% were judged to have another possible or known medical illness which could account for the severe fatigue, and the remaining 45% did not meet the case definition because of histories of psychiatric disorders preceding the onset of fatigue. Minimum prevalence rates for the period 1 September 1989 to 1 September 1991 ranged from 2.0 to 7.3 per 100,000 of the general population across the four study sites and rates based on prorated data ranged from 4.6 to 11.3 per 100,000. More than 80% of the CFS cases were female, most were white, and their average age at onset was approximately 30 years.

Injuries and poisonings in out-of-home child care and home care.

As part of a national telephone survey regarding health events associated with out-of-home child care, data regarding poisonings and injuries were collected. Of 171 reported poisonings, none occurred during out-of-home child care. The rate of injury during out-of-home child care was 1.69 per 100,000 child-hours compared with 2.66 for home care. Overall injury rates were slightly higher for children who attended out-of-home child care than for those who do not. This occurred because children who attended out-of-home child care had a higher injury rate during home care than did the children who did not attend out-of-home child care at all. Although out-of-home child care may carry an increased risk of infectious disease relative to home care, it does not appear to carry an increased risk of injury and, in fact, may confer a lower risk.

Creutzfeldt-Jakob disease in pituitary growth hormone recipients in the United States.

To assess the magnitude of Creutzfeldt-Jakob disease (CJD) occurrence among recipients of pituitary-derived human growth hormone (HGH), we conducted an epidemiologic follow-up of 6284 recipients of HGH distributed through the National Hormone and Pituitary Program. Seven neuropathologically confirmed cases of CJD have occurred in this population to date: six patients with clinical CJD presented with ataxia and imbalance, rather than with altered mentation, which is the most common initial manifestation in sporadic CJD, and one patient died in the preclinical incubation state of the disease. All seven cases occurred among the nearly 700 HGH recipients who started therapy before 1970. Since only 10% of the cohort has been followed up for the 15-year average incubation interval from midpoint of HGH treatment to onset of symptoms, the great majority of potentially exposed patients have not yet attained the requisite incubation period for expression of CJD. The median duration of HGH therapy of 100 months in the CJD cases was significantly longer than 41 months for all patients starting treatment before 1970; thus, the duration of pituitary HGH therapy is a major risk factor for CJD.

Risk of respiratory illness associated with day-care attendance: a nationwide study.

The risk of respiratory and other illnesses in children (age groups: 6 weeks through 17 months, 18 through 35 months, and 36 through 59 months) in various types of day-care facilities was studied. Children considered exposed to day care were those who were enrolled in day care with at least one unrelated child for at least 10 hours per week in each of the 4 weeks before the interview; unexposed children were not enrolled in any regular child care with unrelated children and did not have siblings younger than 5 years of age receiving regular care with unrelated children. Although an increased risk of respiratory illness was associated with attending day care for children in all three age groups, this risk was statistically significant only for children 6 weeks through 17 months of age (odds ratio = 1.6; 95% confidence interval = 1.1 to 2.4) and children 18 through 35 months of age who had no older siblings (odds ratio = 3.4; 95% confidence interval = 2.0 to 6.0). In contrast, day-care attendance was not associated with an increased risk of respiratory illness in children 18 through 35 months of age with older siblings (odds ratio = 1.0). For children aged 6 weeks through 17 months, the exposure to older siblings was associated with an increased risk of respiratory illness; however, for children aged 36 through 59 months, older siblings were protective against respiratory illness. In addition, for the children in each age group currently in day care, increased duration of past exposure to day care was associated with a decreased risk of respiratory illness.(ABSTRACT TRUNCATED AT 250 WORDS)

Status report on the US human growth hormone recipient follow-up study.

Three reported cases of Creutzfeldt-Jakob disease (CJD) in young adults who had received human growth hormone (hGH) raised concerns that pituitary-derived GH had been contaminated. Subsequently reported cases have confirmed this suspicion. The US Public Health Service is conducting an investigation to determine the extent of the problem of CJD in recipients of National Hormone Pituitary Program (NHPP) GH. In addition, other possible adverse effects of GH use including leukemia are being investigated. The design, conduct and current status of the study are the subject of this report. Interview data are now available on 5,240 of the 6,284 subjects treated by the NHPP for growth problems. Analysis is underway.

Exercise testing in the evaluation of human responses to powerline frequency fields.

The present study used exercise testing to examine the effects of 60-Hz electric and magnetic field exposure. Eleven males, 21-29 years, were tested during four experimental sessions (counterbalanced order and double-blind design). These included either 45-min exercise (50% of VO2 max) or no-exercise periods followed by either real field (9-kV/m, 16-A/m) or sham exposure for 2 h in a 60-Hz human exposure facility. Exercise produced a decrease in plasma volume and increases (p less than 0.05) in lactic acid, cortisol, growth hormone and testosterone levels, but these were not different under real and sham field conditions during the 2-h recovery periods. During no-exercise sessions, cardiac interbeat interval was increased (p less than 0.05), i.e., heart rate was slower, when subjects were exposed to real fields. Our results suggest that future studies should focus on evaluation of the effects of 60-Hz fields on the entire process of exercise-induced activation and recovery.

Reye's syndrome and aspirin. Evidence for a dose-response effect.

Data collected from the Public Health Service Main Study of Reye's Syndrome and Medications were analyzed to assess the relationship between the development of Reye's syndrome and the dose of aspirin received during the antecedent respiratory or chickenpox illness. Among those exposed to aspirin, case-patients were found to have received greater average daily and maximum daily doses of aspirin and greater doses of aspirin on the first four days of the antecedent illness (median, 25.1 mg/kg; 33.0 mg/kg; and 65.4 mg/kg; respectively) than did controls (median, 14.5 mg/kg; 19.0 mg/kg; and 27.0 mg/kg; respectively). The excess risk associated with increasing aspirin doses was due primarily to intermediate levels of dose (eg, 15 to 27 mg/kg per day) rather than higher levels (greater than 27 mg/kg per day). The dose difference between exposed case-patients and controls was greatest on days 3 and 4 of the antecedent illness.

Public Health Service study of Reye's syndrome and medications. Report of the main study.

Between January 1985 and May 1986, following completion of a pilot study, a main study concerning the possible association between Reye's syndrome and salicylates was conducted. Twenty-seven patients with stage II or deeper Reye's syndrome whose diagnoses were confirmed by an expert panel and who had appropriate antecedent illnesses (chickenpox, respiratory illness, or gastrointestinal illness) prior to the onset of Reye's syndrome were compared with 140 controls matched for age, race (black or not black), and type and timing of onset of antecedent illness. Controls were selected from the same hospital, emergency room, or school as case-patients or were identified by random-digit telephone dialing. As in the pilot study, a strong statistical association with ingestion of salicylates during the antecedent illness and prior to the onset of Reye's syndrome was observed (odds ratio, 40; lower 95% confidence limit, 5.8). Analysis of the independent risk of aspirin and nonaspirin salicylates revealed a significant association with aspirin (odds ratio, 26; lower 95% confidence limit, 6.4); the independent risk of nonaspirin salicylates could not be assessed because only two cases were not exposed to aspirin. Assessment of epidemiologic issues of concern, including case-control differences in the severity of the antecedent illness, did not explain the high odds ratios that were observed. The high percentage of patients with Reye's syndrome exposed to salicylates (greater than or equal to 90%) in this and prior studies suggests that, though the reported incidence of Reye's syndrome has declined in recent years, concomitant with a decline in salicylate use among children, a majority of Reye's syndrome cases may be attributable to salicylate use.

A microcomputer system for long term monitoring, collecting, analyzing and reporting noninvasive oxygen saturation.

The characteristics of a microcomputer based system for long term monitoring, collecting, analyzing and reporting oxygen saturation are described. Every two seconds SaO2 is transmitted to the microcomputer via the serial port of the oximeter and verified by the program. The microcomputer's real time display and internal data structures are updated. At the end of the data collection period, accumulated data is stored to disk. Report generation modules calculate summary statistics and generate graphs depicting the cumulative time distribution of SaO2, the frequency distribution of SaO2, and an hour by hour representation of the real time display. This inexpensive microcomputer based system allows accurate collection and timely presentation of oxygen saturation levels with monitoring periods of any length replacing tedius and time consuming manual recording, digitizing and analysis.

The pathophysiologic and roentgenologic effects of chest irradiation in breast carcinoma.

We studied the effects of radiation therapy on lung function in 21 patients who underwent modified radical mastectomy and radiotherapy for breast carcinoma. The patients had pulmonary function studies and chest X rays prior to radiation therapy and at six weeks, and three, six, and twelve months following radiation therapy. All pulmonary function studies showed a small but statistically significant decline within the first three months following radiation therapy. Changes in the moderate and large airways continued to occur after three months. None of the changes in pulmonary function were reversible. Radiographic changes occurred in 12 patients. These changes occurred later than the lung function changes (median time for the changes was twelve months) and were unrelated to changes in lung function.

Individual differences in state-dependent retrieval effects of alcohol intoxication.

Individual differences in susceptibility to the state-dependent retrieval (SDR) effects from alcohol in twelve subjects was tested on repeated occasions. There was a threefold variability in frequency of SDR among the subjects. A significant positive correlation between frequency of SDR and history of blackouts and heavy drinking was found.