Rapid gene fusion testing using the NanoString nCounter platform to improve pediatric leukemia diagnoses in Sub-Saharan Africa.

Risk stratification and molecular targeting have been key to increasing cure rates for pediatric cancers in high-income countries. In contrast, precise diagnosis in low-resource settings is hindered by insufficient pathology infrastructure. The Global HOPE program aims to improve outcomes for pediatric cancer in Sub-Saharan Africa (SSA) by building local clinical care and diagnostic capacity. This study aimed to assess the feasibility of implementing molecular assays to improve leukemia diagnoses in SSA. Custom NanoString nCounter gene fusion assays, previously validated in the US, were used to test samples from suspected leukemia patients. The NanoString platform was chosen due to relatively low cost, minimal technical and bioinformatics expertise required, ability to test sub-optimal RNA, and rapid turnaround time. Fusion results were analyzed blindly, then compared to morphology and flow cytometry results. Of 117 leukemia samples, 74 were fusion-positive, 30 were negative, 7 were not interpretable, and 6 failed RNA quality. Nine additional samples were negative for leukemia by flow cytometry and negative for gene fusions. All 74 gene fusions aligned with the immunophenotype determined by flow cytometry. Fourteen samples had additional information available to further confirm the accuracy of the gene fusion results. The testing provided a more precise diagnosis in >60% of cases, and 9 cases were identified that could be treated with an available tyrosine kinase inhibitor, if detected at diagnosis. As risk-stratified and targeted therapies become more available in SSA, implementing this testing in real-time will enable the treatment of pediatric cancer to move toward incorporating risk stratification for optimized therapy.

Simple kcal/kg formula is comparable to prediction equations for estimating resting energy expenditure in older cognitively impaired long term care residents.

OBJECTIVE: Assessment of energy needs is a critical step in developing the nutrition care plan, especially for individuals unable to modulate their own energy intakes. The purpose of this study was to assess precision and accuracy of commonly used prediction equations in comparison to measured resting energy expenditure in a sample of "oldest old" adults residing in long term care (LTC). SUBJECTS AND DESIGN: Resting energy expenditure (mREE) was measured by indirect calorimetry in 45 residents aged 86.1 ± 7.3 years, and compared to frequently used prediction equations (pREE): Mifflin St.Jeor, Harris Benedict, World Health Organization and Owen. Precision and accuracy were determined by concordance correlation coefficients and number of individuals within ± 10% of mREE. Bland Altman plots with linear dependence trends were constructed to visualize agreement. To complete analyses, the common 25 kcal/kg formula was assessed and alternative formulas were determined for best fit by regressing adjusted mREE on body weight. RESULTS: mREE averaged 976.2 ± 190.3 kcal/day for females and 1260.0 ± 275.9 kcal/d for males. The strength of the relationships between pREE and mREE were only moderate (r = 0.41 - 0.72). In examining linear trends in the Bland Altman plots, significant systematic deviation from mREE was detected for all pREE. Two kcal/kg formulas were generated: 20.6 kcal/kg for females and 22.7 kcal/kg for males, which were not significantly different. CONCLUSION: None of the prediction equations adequately estimated energy needs in this sample of the "oldest old." A simple formula using 21-23 kcal/kg may be a more practical and reliable method to determine energy needs in the LTC setting.

Monitoring autophagy in glioblastoma with antibody against isoform B of human microtubule-associated protein 1 light chain 3.

Autophagy, an evolutionarily conserved response to stress, has recently been implicated in cancer initiation and progression, but the detailed mechanisms and functions have not yet been fully elucidated. One major obstacle to our understanding is lack of an efficient and robust method to specifically monitor autophagic cells in cancer specimens. To identify molecular events associated with autophagy, we performed cDNA microarray analysis of autophagic glioblastoma cell lines. Based on the analysis, we raised a polyclonal antibody against isoform B of human microtubule-associated protein 1 light chain 3 (LC3B). Application of the anti-LC3B antibody revealed the presence of autophagic cells in both in vitro and in vivo settings. Of the 65 glioblastoma tissues, 31 had highly positive cytoplasmic staining of LC3B. The statistical interaction between cytoplasmic staining of LC3B and Karnofsky Performance Scale score was significant. High expression of LC3B was associated with an improved outcome for patients with poorer performance, whereas, for patients with normal performance, survival was better for patients with low staining than with high staining of LC3B. Anti-LC3B antibody provides a useful tool for monitoring the induction of autophagy in cancer cells and tissues.

Ionizing radiation induces apoptosis and inhibits neuronal differentiation in rat neural stem cells via the c-Jun NH2-terminal kinase (JNK) pathway.

A substantial number of neural stem cells (NSCs) continue to proliferate and generate neurons in the central nervous system throughout life. Ionizing radiation, an important adjuvant therapy for glioma patients, may damage NSCs and cause neuronal deficits, such as cognitive dysfunction and memory impairment. However, the precise mechanism of radiation effects on death and differentiation of NSCs remains largely unknown. Here, we found that radiation induced apoptosis in NSCs via the mitochondrial pathway, upregulating the ratio of Bax to Bcl-2 and releasing cytochrome c into the cytoplasm. Radiation also inhibited neuronal differentiation of NSCs by 50%. Of the three stress-associated mitogen-activated protein kinases (MAPKs), only c-Jun NH(2)-terminal kinase (JNK) was activated in NSCs after radiation. Interestingly, JNK inhibition by the specific inhibitor SP600125 rescued NSCs from apoptosis and improved neuronal differentiation. Furthermore, we examined whether radiation directly inhibits neuronal differentiation or not. Radiation did not affect the promoter activity of NeuroD, a basic helix-loop-helix transcription factor that regulates the expression of neuronal differentiation markers. Radiation induced more apoptosis in NeuroD-positive cells than NeuroD-negative cells. We concluded that radiation activates JNK and induces apoptosis, especially in neural progenitor cells, resulting in the inhibition of neurogenesis. Our findings raise the possibility that JNK inhibition has therapeutic potential in protecting NSCs from the adverse effects of radiation.

Work and social support: a comparison of consumers who have achieved stability in ACT and clubhouse programs.

A current debate in the field is whether consumers, who have achieved stability in Assertive Community Treatment programs, can be transferred to less intensive services. To bring some data to bear on this question, this study compared consumers and members, who have achieved stability, in either an Assertive Community Treatment (ACT) or a clubhouse program, on domains of vocational activity, social relationships/loneliness and community integration. The 51 stable clients from the two programs who were interviewed, reported similar vocational activity, similar experiences with social relationships and social networks, and similar community integration. Clients in both groups were less lonely than previously reported in the literature. Study results indicate, that for those clients who have achieved stability, there are sufficient similarities between consumers in the two programs, to suggest a potential for movement from more to less intensive programs with less disruption than previously assumed possible.

Mental health expenditures for services for people with severe mental illnesses.

OBJECTIVE: To provide comprehensive information on expenditures for mental health and substance abuse services for a large number of people with severe mental illnesses, this study examined use of major types of clinical-medical mental health and psychiatric rehabilitation services over a one-year period. METHODS: Data were obtained for 1,890 clients in ten public county-based nonmetropolitan mental health systems in Wisconsin. Expenditures were for services provided with public funding, including local sources of funding, Medicaid, and Medicare. Data about services and expenditures were obtained from county records and unduplicated Medicaid claims for 12 months in 1989 and 1990. RESULTS: Expenditures per client averaged $10,995 for one year ($13,992 in 1994 dollars), with a maximum of $95,093. Expenditures for community-based outpatient services, including residential care and vocational services, represented 53.5 percent of all expenditures; residential care accounted for 12.4 percent and vocational services for 5.7 percent. Overall, 46.5 percent was spent for institutional care, with inpatient hospital care accounting for 12.6 percent. Approximately 40.6 percent of total expenditures were for services not typically covered under managed care plans. CONCLUSIONS: Expenditures for community-based care accounted for more than half of total expenditures. Expenditure patterns revealed the important role of social and rehabilitation services, a role that must be continued in managed care arrangements if they are to provide adequate services for people with severe mental illnesses.

Services for clients of community support programs in rural Wisconsin.

To improve understanding of services provided or coordinated by rural community support programs (CSPs) for people with severe mental illness, this article identifies services most used by clients and the amounts of services used. Data on publicly funded services for more than 900 clients in 13 rural CSPs in a midwestern state have been analyzed. Virtually all clients were Caucasian. Information about types and amounts of client services for 12 consecutive months was obtained from county information systems, local records, and Medicaid claims. Most CSP clients use case management, community support, medication checks, counseling, and medication counseling services. Much smaller percentages use other outpatient, residential, vocational, and inpatient services. Significant amounts of only two services, case management and community support, are reported. The findings emphasize the ability of rural mental health providers to supply general services, but some limitation in provision of specialized services and facilities.

Pharmacological properties of 403U76, a new chemical class of 5-hydroxytryptamine- and noradrenaline-reuptake inhibitor.

403U76 (5-chloro-[[2-[(dimethylamino)methyl]phenyl]thio]benzene- methanol hydrochloride) is a potent, competitive, inhibitor of 5-hydroxytryptamine (5-HT) and noradenaline reuptake into rat brain synaptosomes. Inhibition of 5-HT uptake in-vivo by 403U76 was demonstrated by potentiation of the behavioural effects of 5-hydroxytryptophan in rats and mice and blockade of p-induced depletion of 5-HT in rats. The firing of 5-HT-ergic dorsal raphe neurons in rats was decreased after intravenous administration of low doses of 403U76 as would be predicted for a 5-HT uptake inhibitor. 403U76 antagonized tetrabenazine-induced sedation, an effect associated with inhibitors of noradrenaline uptake, but not with inhibitors of 5-HT uptake. Thus 403U76 affects noradrenergic as well as 5-HT-ergic neurotransmission in-vivo. Potential anxiolytic activity was indicated by reductions in isolation-induced vocalizations in neonates after 403U76 treatment. Low intravenous doses of 403U76 were well tolerated and had no sustained cardiovascular effects. There were no deleterious behavioural side-effects at active doses. Effects observed on isolated tissues or transmitter receptors occurred only at very high concentrations and were pharmacologically unimportant. Thus 403U76 can be considered a potential antidepressant/anxiolytic agent that is a potent, selective inhibitor of 5-HT and noradrenaline reuptake.

Use of Medicaid for mental health care by clients of community support programs.

The use of Medicaid for mental health services by the severely mentally ill is examined using paid Medicaid claims for clients of Community Support Programs in Wisconsin. The extent to which clients participate in Medicaid, the types and amounts of mental health services, and the costs of services are discussed. A narrow majority of CSP clients use Medicaid, which provides a substantial amount of case management service for program participants. Psychotropic medications are also used by a large percentage of clients. Other relatively common services covered by Medicaid are medication checks and psychotherapy. Average annual Medicaid payments for mental health services (including medications) are $2438.

Staffing patterns in rural community support programs.

OBJECTIVE AND METHODS: Rural mental health programs are thought to have difficulty recruiting and retaining qualified staff and to have excessively large caseloads per staff member. The authors surveyed 12 community support programs in small cities and rural areas of Wisconsin to determine staff size and characteristics, staff-to-client ratios, and the relative use of paraprofessional staff and professional staff, excluding psychiatrists. RESULTS: The programs surveyed had an average of 5.6 full-time-equivalent caregiving staff and an average caregiver-to-client ratio of 1 to 13. Caregiving staff were predominantly female, had been in the mental health field a mean of 8.8 years, and received an average salary of $20,732. Although program directors indicated that about one-fifth of staff left in the previous year, they reported little difficulty recruiting staff. Thirty-eight percent of all caregivers were paraprofessionals; in several programs, more than half the caregivers were paraprofessionals. Use of paraprofessionals is more common in rural programs; thus caregivers in those programs are likely to have lower levels of salary and experience. CONCLUSIONS: The community support programs in the sample employed experienced caregivers and had staff-to-client ratios that were close to the ratio recommended for such programs by Wisconsin legislation. The survey findings suggest that claims of acute staffing problems in rural community support programs need to be examined more closely, with attention to the relationship between staff characteristics and client outcomes.

Conformationally restricted analogues of remoxipride as potential antipsychotic agents.

Several conformationally restricted derivatives of (S)-3-bromo-N-((1-ethyl-2-pyrrolidinyl)methyl)-2,6-dimethoxybenzamide (remoxipride) were synthesized and evaluated in vitro for their ability to inhibit [3H]raclopride binding at the dopamine D-2 receptor. The cyclic benzamides designed to mimic the intramolecular hydrogen bonding of desmethylremoxipride (4, FLA-797) included 2,3-dihydro-4H-1,3-benzoxazin-4-ones, 2,3-dihydro-4H-1,3-benzthiazin-4-ones, phthalimides, 1-isoindolinones, 1,2-benzisothiazol-3(2H)-ones, and 1,2-benzisothiazol-3(2H)-one 1,1-dioxides. In this series, enhanced affinities to the dopamine D-2 receptor were not observed. The phthalimidine analogue 24b ((S)-6-chloro-2-(1-ethylpyrrolidinyl)-1-isoindolinone) exhibited the highest affinity to the dopamine D-2 receptor with an IC50 of 1.3 microM, which was equipotent to remoxipride.

Synthesis and evaluation of the antidepressant activity of the enantiomers of bupropion.

The synthesis of the enantiomers of bupropion, (rac)-2-tert-butylamino-3'-chloropropiophenone 1 (Wellbutrin) is described. The enantiomers were compared with the racemate in both the tetrabenazine-induced sedation model and the inhibition of uptake of biogenic amine assay. No significant differences were found in their potencies to reverse tetrabenazine-induced sedation in mice or in their IC50 values as inhibitors of biogenic amine uptake into nerve endings obtained from mouse brain.

Alterations in extracellular and tissue levels of biogenic amines in rat brain induced by the serotonin2 receptor antagonist, ritanserin.

Systemic administration of ritanserin elicited rapid changes in dopamine (DA) and serotonin (5-HT) levels in both dialysate and neuronal tissue extracts. These effects occurred in both a site-selective and a dose-related manner. Increases in extracellular levels of DA and 5-HT in the nucleus accumbens were maximal at 120-140 min after treatment. A dose of 0.63 mg/kg of ritanserin elicited larger and more prolonged increases in extracellular DA and 5-HT levels than did the 0.3 mg/kg dose. By contrast, 0.63 mg/kg of ritanserin elicited no changes in either DA or 5-HT levels with dialysate collected from the striatum. Ritanserin also induced dose-related decreases in tissue levels of DA and 5-HT from the nucleus accumbens. The site specificity of action was again noted in that there were no dose-dependent decreases in tissue levels of DA or 5-HT measured from the striatum. Ritanserin exerted little effect on metabolite levels from either dialysate or tissue extracts. Taken together, these findings show that selective 5-HT2 receptor antagonism modulates DA and 5-HT neurotransmission in a specific manner. These actions appear to involve increased release of DA and 5-HT rather than significant changes in metabolism. These findings add further weight to the importance of 5-HT2 receptor interactions as an important component of antipsychotic activity.

Falling through the cracks: care of the chronically mentally ill in the United States, Germany, and the United Kingdom.

In comparing the development and strength of community-based services for the chronically mentally ill in the United States, Germany, and the United Kingdom, I analyze how the structure of each country's general medical system has influenced services for the chronically mentally ill and the extent to which more universal medical care systems are associated with stronger community-based systems for the mentally ill. Community-based services are frail and inadequate in all three countries, in each country for different reasons. The specifics of organization of the health care system seem less important in shaping these outcomes than the status of mental health care as a national priority.

Analysis of rat brain microdialysate by gas chromatography-high-resolution selected-ion monitoring mass spectrometry.

Gas chromatography-high-resolution selected-ion monitoring mass spectrometry was used to analyze catecholamine metabolites in rat brain microdialysate. Dialysate samples were collected in vials containing stable isotope analogues of homovanillic acid (HVA), 3-methoxy-4-hydroxyphenylglycol (MHPG) and 5-hydroxyindoleacetic acid (5HIAA) and analyzed as their trimethylsilyl derivatives. The metabolite levels were monitored at 20-min intervals throughout the time course of the experiment, beginning immediately after surgery and implantation of the dialysis probe and ending 4 h after amphetamine treatment. The levels of HVA were observed to decrease after amphetamine treatment, while those of MHPG and 5HIAA did not change significantly.

Effects of the 5-HT2 receptor antagonist, ritanserin, on biogenic amines in the rat nucleus accumbens.

The effects of acute ritanserin treatment on dopamine (DA) and serotonin (5-HT) release and metabolism were studied in tissue and microdialysis samples of the nucleus accumbens in rats. Administration of a moderate dose of ritanserin elicited decreases in DA and 5-HT in tissue with concomitant increases in extracellular fluid. These data show that selective 5-HT2 receptor antagonism modulates DA as well as 5-HT neurotransmission and adds support to the suggestion that 5-HT/DA interactions may be important in the treatment of psychoses.