Enhanced CRFR1-Dependent Regulation of a Ventral Tegmental Area to Prelimbic Cortex Projection Establishes Susceptibility to Stress-Induced Cocaine Seeking.

The ability of stress to trigger cocaine seeking in humans and rodents is variable and is determined by the amount and pattern of prior drug use. This study examined the role of a corticotropin releasing factor (CRF)-regulated dopaminergic projection from the ventral tegmental area (VTA) to the prelimbic cortex in shock-induced cocaine seeking and its recruitment under self-administration conditions that establish relapse vulnerability. Male rats with a history of daily long-access (LgA; 14 × 6 h/d) but not short-access (ShA; 14 × 2 h/d) self-administration showed robust shock-induced cocaine seeking. This was associated with a heightened shock-induced prelimbic cortex Fos response and activation of cholera toxin b retro-labeled VTA neurons that project to the prelimbic cortex. Chemogenetic inhibition of this pathway using a dual virus intersectional hM4Di DREADD (designer receptor exclusively activated by designer drug) based approach prevented shock-induced cocaine seeking. Both shock-induced reinstatement and the prelimbic cortex Fos response were prevented by bilateral intra-VTA injections of the CRF receptor 1 (CRFR1) antagonist, antalarmin. Moreover, pharmacological disconnection of the CRF-regulated dopaminergic projection to the prelimbic cortex by injection of antalarmin into the VTA in one hemisphere and the D1 receptor antagonist, SCH23390, into the prelimbic cortex of the contralateral hemisphere prevented shock-induced cocaine seeking. Finally, LgA, but not ShA, cocaine self-administration resulted in increased VTA CRFR1 mRNA levels as measured using in situ hybridization. Altogether, these findings suggest that excessive cocaine use may establish susceptibility to stress-induced relapse by recruiting CRF regulation of a stressor-responsive mesocortical dopaminergic pathway.SIGNIFICANCE STATEMENT Understanding the neural pathways and mechanisms through which stress triggers relapse to cocaine use is critical for the development of more effective treatment approaches. Prior work has demonstrated a critical role for the neuropeptide corticotropin releasing factor (CRF) in stress-induced cocaine seeking. Here we provide evidence that stress-induced reinstatement in a rat model of relapse is mediated by a CRF-regulated dopaminergic projection from the ventral tegmental area (VTA) that activates dopamine D1 receptors in the prelimbic cortex. Moreover, we report that this pathway may be recruited as a result of daily cocaine self-administration under conditions of extended drug access/heightened drug intake, likely as a result of increased CRFR1 expression in the VTA, thereby promoting susceptibility to stress-induced cocaine seeking.

Reversal of defective lysosomal transport in NPC disease ameliorates liver dysfunction and neurodegeneration in the npc1-/- mouse.

Niemann-Pick type C disease is largely attributable to an inactivating mutation of NPC1 protein, which normally aids movement of unesterified cholesterol (C) from the endosomal/lysosomal (E/L) compartment to the cytosolic compartment of cells throughout the body. This defect results in activation of macrophages in many tissues, progressive liver disease, and neurodegeneration. In the npc1(-/-) mouse, a model of this disease, the whole-animal C pool expands from 2,082 to 4,925 mg/kg body weight (bw) and the hepatic C pool increases from 132 to 1,485 mg/kg bw between birth and 49 days of age. A single dose of 2-hydroxypropyl-beta-cyclodextrin (CYCLO) administered at 7 days of age immediately caused this sequestered C to flow from the lysosomes to the cytosolic pool in many organs, resulting in a marked increase in cholesteryl esters, suppression of C but not fatty acid synthesis, down-regulation of genes controlled by sterol regulatory element 2, and up-regulation of many liver X receptor target genes. There was also decreased expression of proinflammatory proteins in the liver and brain. In the liver, where the rate of C sequestration equaled 79 mg x d(-1) x kg(-1), treatment with CYCLO within 24 h increased C movement out of the E/L compartment from near 0 to 233 mg x d(-1) x kg(-1). By 49 days of age, this single injection of CYCLO resulted in a reduction in whole-body C burden of >900 mg/kg, marked improvement in liver function tests, much less neurodegeneration, and, ultimately, significant prolongation of life. These findings suggest that CYCLO acutely reverses the lysosomal transport defect seen in NPC disease.

Cyclodextrin overcomes the transport defect in nearly every organ of NPC1 mice leading to excretion of sequestered cholesterol as bile acid.

A mutation in NPC1 leads to sequestration of unesterified cholesterol in the late endosomal/lysosomal compartment of every cell culminating in the development of pulmonary, hepatic, and neurodegenerative disease. Acute administration of 2-hydroxypropyl-beta-cyclodextrin (CYCLO) rapidly overcomes this transport defect in both the 7-day-old pup and 49-day-old mature npc1(-/-) mouse, even though this compound is cleared from the body and plasma six times faster in the mature mouse than in the neonatal animal. The liberated cholesterol flows into the cytosolic ester pool, suppresses sterol synthesis, down-regulates SREBP2 and its target genes, and reduces expression of macrophage-associated inflammatory genes. These effects are seen in the liver and brain, as well as in peripheral organs like the spleen and kidney. Only the lung appears to be resistant to these effects. Forty-eight h after CYCLO administration to the 49-day-old animals, fecal acidic, but not neutral, sterol output increases, whole-animal cholesterol burden is reduced, and the hepatic and neurological inflammation is ameliorated. However, lifespan is extended only when the CYCLO is administered to the 7-day-old animals. These studies demonstrate that CYCLO administration acutely reverses the cholesterol transport defect seen in the NPC1 mouse at any age, and this reversal allows the sequestered sterol to be excreted from the body as bile acid.

Case-control comparison of quality of life in long-term ovarian germ cell tumor survivors: a gynecologic oncology group study.

Ovarian germ cell cancer is a rare tumor. Approximately 1000 to 2000 women in the United States will be diagnosed with ovarian germ cell cancer in 2007. When it occurs, it is usually diagnosed before age 20 and is highly responsive to therapy. Most patients live a full life span. The 5-year relative survival rate is 95%. This article describes differences in quality of life issues between ovarian germ cell cancer survivors and young women who have not experienced cancer and were matched to survivors on age, education, and race (by the acquaintance control method). Survivors and controls completed mail and phone surveys. A multivariable logistic regression model was adjusted for age, education, household income, marital status, and perception of fertility. Compared to controls, germ cell cancer survivors expressed more reproductive concerns and reported worse sexual functioning, but they also experienced greater appreciation of life and more affective (i.e., emotional) social support. Future research is suggested to test interventions to enhance quality of life for ovarian germ cell cancer survivors in the areas of sexual functioning and reproductive concerns, but only for survivors who are in distress or in need of the support. Potential screening questions are offered for clinicians, but further research is needed to assess their validity as screening tools.

Profiting from Probability; Combining Low and High Probability Isotopes as a Tool Extending the Dynamic Range of an Assay Measuring Amphetamine and Methamphetamine in Urine.

A wide range of concentrations are frequently observed when measuring drugs of abuse in urine toxicology samples; this is especially true for amphetamine and methamphetamine. Routine liquid chromatography-tandem mass spectrometry confirmatory methods commonly anchored at a 50 ng/mL lower limit of quantitation can span approximately a 100-fold concentration range before regions of non-linearity are reached deteriorating accurate quantitation and qualitative assessments. In our experience, approximately a quarter of amphetamine and methamphetamine positive samples are above a 5,000 ng/mL upper limit of quantitation and thus require reanalysis with dilution for accurate quantitative and acceptable qualitative results. We present here the development of an analytical method capable of accurately quantifying samples with concentrations spanning several orders of magnitude without the need for sample dilution and reanalysis. For each analyte the major isotopes were monitored for analysis through the lower concentration ranges (50-5,000 ng/mL), and the naturally occurring, low probability 13C2 isotopes were monitored for the analysis of the high concentration samples (5,000-100,000 ng/mL amphetamine and 5,000-200,000 ng/mL methamphetamine). The method simultaneously monitors transitions for the molecules containing only 12C and 13C2 isotopologues eliminating the need for re-extraction and reanalysis of high concentration samples.

Chronic physical effects and health care utilization in long-term ovarian germ cell tumor survivors: a Gynecologic Oncology Group study.

PURPOSE: This study compares late effects of treatment on physical well-being and utilization of health care resources between ovarian germ cell tumor (OGCT) survivors and age/race/education-matched controls. PATIENTS AND METHODS: Eligible patients had OGCT treated with surgery and chemotherapy and were disease-free for at least 2 years at time of enrollment. The matched control group was selected from acquaintances recommended by survivors. Symptoms and function were measured using previously validated scales. Health care utilization was assessed by questions regarding health insurance coverage and health services utilization. RESULTS: One hundred thirty-two survivors and 137 controls completed the study. Survivors were significantly more likely to report a diagnosis of hypertension (17% v 8%, P = .02), and marginally hypercholesterolemia (9.8% v 4.4%, P = .09), and hearing loss (5.3% v 1.5%, P = .09) compared with controls. There were no significant differences in the rates of self-reported arthritis, heart, pulmonary or kidney disease, diabetes, non-OGCT malignancies, anxiety, hearing loss, or eating disorders between groups. Among chronic functional problems, numbness, tinnitus, nausea elicited by reminders of chemotherapy (v general nausea triggers for controls), and Raynaud's symptoms were reported more frequently by survivors. Patients who received vincristine, dactinomycin, and cyclophosphamide in addition to cisplatin therapy had increased functional complaints, particularly numbness and nausea. Health care utilization was similar, but 15.9% of survivors reported being denied health insurance versus 4.4% of controls (P < .001). CONCLUSION: Although a few sequelae of treatment persist, in general, OGCT survivors enjoy a healthy life comparable to that of controls.

Reproductive and sexual function after platinum-based chemotherapy in long-term ovarian germ cell tumor survivors: a Gynecologic Oncology Group Study.

PURPOSE: To compare malignant ovarian germ cell tumor survivors with a matched control group of females on menstrual and reproductive outcomes, sexual functioning, and dyadic adjustment. PATIENTS AND METHODS: Eligible patients met the following criteria: (1) history of malignant ovarian germ cell tumor; (2) treatment with surgery plus platinum-based chemotherapy; (3) age at least 18 years and continuously disease-free with minimum follow-up of 2 years; (4) capability of completing questionnaire and telephone interview; and (5) completion of written informed consent. The control group was drawn from acquaintances recommended by survivors and matched for age, race, and education. Scales with established reliability and validity were used to measure quality-of-life concepts of sexual functioning and social networks. RESULTS: One hundred thirty-two survivors and 137 controls completed the study. Of 132 survivors, 71 (53.8%) had fertility-sparing surgery. Of fertile survivors, 62 (87.3%) reported still having menstrual periods. Twenty-four survivors reported 37 offspring after cancer treatment. Compared with controls, survivors had significantly greater reproductive concerns (P < .0001), less sexual pleasure (P = .003), and lower scores on the total Sexual Activity Scale Score (P = .001). However, survivors had better dyadic consensus (P = .004), dyadic satisfaction (P = .005), and dyadic cohesion (P = .014). CONCLUSION: Women who had fertility-sparing surgery were very likely to retain menstrual function and fertility after chemotherapy. Although there is some increase in gynecologic symptoms and diminution in sexual pleasure, survivors tended to have stronger, more positive relationships with significant others.

Quality of life after among ovarian germ cell cancer survivors: a narrative analysis.

PURPOSE/OBJECTIVES: To describe and interpret the meaning of experiences that are important to the quality of living of ovarian germ cell cancer survivors. RESEARCH APPROACH: Qualitative description within a constructivist paradigm. SETTING: 32 member sites of the Gynecologic Oncology Group and the University of Texas M.D. Anderson Cancer Center in Houston. PARTICIPANTS: 109 women between the ages of 19-64 (median age = 36) who were enrolled on prospective clinical trials of cisplatin-based chemotherapy after surgery and disease-free for at least two years. METHODOLOGIC APPROACH: As part of a larger study, narrative responses to four semistructured questions were collected at the end of a telephone interview. Using naturalistic inquiry and qualitative description techniques, content labels were assigned to units of text that seemed to encapsulate one complete thought or idea. The labeled groups were collapsed into interpreted subthemes. Finally, four general themes were constructed as representations of shared narrative responses and meanings. FINDINGS: The four constructed themes are celebrating illness, experiencing empathetic affirmation, mourning losses, and valuing illness. INTERPRETATION: In addition to measuring physical, psychological, and sexual functioning in women surviving ovarian germ cell cancer, nurses also must understand how these issues fit into their everyday lived experiences. The four themes may help clinicians and researchers to understand issues that are important to the quality of living of ovarian germ cell cancer survivors.