Comparisons of urinary versus serum cortisol in older persons

The present analysis compared urine- versus serum-based amounts of the stress hormone cortisol in two older adult samples, given that urine as a sample medium is a less expensive and less invasive method of determining cortisol amounts relative to serum. Seventy-three older adults provided urine samples as part of an ongoing study to assess levels of cortisol as a function of intellectual efficacy/performance; these data were compared to serum cortisol levels obtained from 96 older adults in a separate study examining health beliefs and cortisol levels. Analyses indicated that the cortisol levels did not differ across samples, i.e., cortisol amounts measured in serum or urine yielded similar, typical (within normal ranges) results. The data, though preliminary, indicate that urine may provide an alternative to serum when assessing cortisol in older persons.     

Serum antibodies to Balamuthia mandrillaris, a free-living amoeba recently demonstrated to cause granulomatous amoebic encephalitis

Free-living amoebae cause three well-defined disease entities: a rapidly fatal primary meningoencephalitis, a chronic granulomatous amoebic encephalitis (GAE), and a chronic amoebic keratitis. GAE occurs in immunocompromised persons. Recently, another type of free-living amoeba, Balamuthia mandrillaris, has been shown to cause GAE. The finding that this amoeba has caused infection in some healthy children has raised the possibility that humans may lack immunity to B. mandrillaris. Human serum was examined for the presence of surface antibodies specific for this amoeba by immunofluorescence. Sera from adults contained titers of 1/64-1/256 of anti-B. mandrillaris antibodies (IgM and IgG classes), which did not cross-react with other amoebae. Cord blood contained very low antibody levels, but levels similar to those in adults were seen in serum of 1- to 5-year-old children.     

Meta-analysis of cue-reactivity in addiction research

Aims. The cue-reactivity procedure exposes addicts to a variety of drug-related stimuli while self-report of craving and physiological responses are monitored. The present review sought to determine the magnitude and overall pattern of responses typically found in cue-reactivity research and which, if any, learning-based model of cue reactivity is best supported by the findings. Design. Meta-analytical techniques were used to select and evaluate results from 41 cue-reactivity studies that compared responses of alcoholics, cigarette smokers, cocaine addicts or heroin addicts to drug-related versus neutral stimuli. Effect sizes were calculated, separately by addict type, for self-report of craving and physiological responses (heart rate, sweat gland activity and skin temperature). Findings. Across all addict groups, the effect size for craving was +0.92. Alcoholics had a significantly smaller craving effect size (+0.53) compared to other addict groups (+1.18 to +1.29). Relatively smaller effect sizes were found for physiological responses. The general profile of effect sizes across all addict groups was increased heart rate (+0.26) and sweat gland activity (+0.40) and decreased skin temperature (-0.24) when addicts were presented with drug-related stimuli. Conclusions. The cuereactivity paradigm can produce a stable profile of significant effects and, therefore, has a number of potential applications for investigating addictive phenomena. The implications of these findings for conditioning-based models of cue-reactivity phenomena are discussed.     

Absorption and disposition of cobalt naphthenate in rats after a single oral dose

The absorption and disposition of inorganic cobalt salts after oral administration have not been completely characterized. The objective of this project was to investigate the absorption and disposition of cobalt naphthenate in Fischer 344 rats following a single oral dose. Cobalt naphthenate was given orally at 3 doses: 0.333, 3.33, or 33.3 mg Co(II)/kg. Tissues, urine, and feces were collected over a 36-h period from the low- and high-dose groups; blood was collected from all 3 dose groups. The majority of the dose in both the low- and high-dose groups was excreted in the feces (42% and 73.1%, respectively), indicating that cobalt was incompletely absorbed from the gastrointestinal tract following oral dosing. The percent of the dose excreted in the urine was similar for low and high doses (31.8% and 26.3%, respectively). Cobalt concentrations were found to be highest in the liver and kidneys. The blood versus time cobalt concentration curves for the low-dose, intermediate-dose, and high-dose groups were elevated 4- to 5-fold, 14- to 25-fold, and 25- to 60-fold over control blood levels, respectively. The peak plasma concentrations of 0.6 and 1.7 microg Co(II)/ml occurred at approximately 4.3 h for the intermediate-dose group, and 3.3 h for the high-dose group. The terminal elimination half-lives were 24.7 and 24 h for the intermediate- and high-dose groups, respectively. Thus, although the extent of cobalt absorption as indicated by the blood concentrations and areas under the blood-time curves was not proportional to dose, the calculated pharmacokinetic values for the time to peak blood concentration and the apparent elimination rate constants were independent of dose. The amount excreted in the urine was also proportional to the dose. These apparent anomalies were not related to protein binding in blood.     

Premenstrual symptoms in women with premenstrual asthma

STUDY OBJECTIVES: To characterize and compare premenstrual symptoms (mood, physical) throughout two menstrual cycles with and without estradiol administration, in women with premenstrual asthma (PMA), and to examine relationships between asthma symptoms versus premenstrual symptoms and pulmonary function versus premenstrual symptoms in these women. DESIGN: Open-label, longitudinal, 9-week study (consisting of two complete menstrual cycles). SETTING: Clinical study at the University of Kentucky; data analysis at the University of British Columbia and Children's and Women's Health Centre of British Columbia. PATIENTS: Fourteen women (age 35.6 +/- 6.6 yrs) with mild to moderate asthma with a baseline ratio of forced expiratory volume in 1 second:forced vital capacity of 0.72 +/- 0.12. INTERVENTIONS: Women were followed for two complete menstrual cycles. During the second complete cycle (i.e., cycle 3), they received estradiol 2 mg orally between days 23 and 28 (premenstrual). MEASUREMENTS AND MAIN RESULTS: Throughout both cycles 2 and 3, each subject recorded premenstrual symptom questionnaire scores (15 mood and physical symptoms, graded 0-3) every morning on awakening. Peak expiratory flow rate (PEFR) and visual analog scales of asthma symptoms (cough, wheezing, breathlessness, chest tightness) were recorded daily at the same time. Seven subjects showed a classic pattern of premenstrual symptoms. Four of the five subjects who complained of PMA symptoms at study enrollment also demonstrated this classic pattern of premenstrual symptoms. After estradiol administration, four women had lower symptom scores, eight had higher scores, and two had the same scores. Overall, estradiol had no significant effect on symptoms (mean area under the curve 18.9 +/- 14.8 day(-1) vs 20.3 +/- 14.8 day(-1), p>0.05). Ten subjects had significant relationships between asthma symptoms and premenstrual symptoms, whereas six had significant relationships between PEFR and premenstrual symptoms. CONCLUSIONS: Exogenous estradiol administration had no significant effect on premenstrual symptoms in women with PMA. The lack of a significant effect allows for patient blinding in a placebo-controlled, crossover study of exogenous estradiol in PMA that is currently under way Clinical implications of relationships between asthma symptoms versus premenstrual symptoms and pulmonary function versus premenstrual symptoms may warrant further study.     

Induction of immunity to prostate cancer antigens: results of a clinical trial of vaccination with irradiated autologous prostate tumor cells engineered to secrete granulocyte-macrophage colony-stimulating factor using ex vivo gene transfer.

Vaccination with irradiated granulocyte-macrophage colony-stimulating factor (GM-CSF)-secreting gene-transduced cancer vaccines induces tumoricidal immune responses. In a Phase I human gene therapy trial, eight immunocompetent prostate cancer (PCA) patients were treated with autologous, GM-CSF-secreting, irradiated tumor vaccines prepared from ex vivo retroviral transduction of surgically harvested cells. Expansion of primary cultures of autologous vaccine cells was successful to meet trial specifications in 8 of 11 cases (73%); the yields of the primary culture cell limited the number of courses of vaccination. Side effects were pruritis, erythema, and swelling at vaccination sites. Vaccine site biopsies manifested infiltrates of dendritic cells and macrophages among prostate tumor vaccine cells. Vaccination activated new T-cell and B-cell immune responses against PCA antigens. T-cell responses, evaluated by assessing delayed-type hypersensitivity (DTH) reactions against untransduced autologous tumor cells, were evident in two of eight patients before vaccination and in seven of eight patients after treatment. Reactive DTH site biopsies manifested infiltrates of effector cells consisting of CD45RO+ T-cells, and degranulating eosinophils consistent with activation of both Th1 and Th2 T-cell responses. A distinctive eosinophilic vasculitis was evident near autologous tumor cells at vaccine sites, and at DTH sites. B-cell responses were also induced. Sera from three of eight vaccinated men contained new antibodies recognizing polypeptides of 26, 31, and 150 kDa in protein extracts from prostate cells. The 150-kDa polypeptide was expressed by LNCaP and PC-3 PCA cells, as well as by normal prostate epithelial cells, but not by prostate stromal cells. No antibodies against prostate-specific antigen were detected. These data suggest that both T-cell and B-cell immune responses to human PCA can be generated by treatment with irradiated, GM-CSF gene-transduced PCA vaccines.     

Identification of a human anti-CD55 single-chain Fv by subtractive panning of a phage library using tumor and nontumor cell lines.

A large naive human single-chain (sc) Fv phage library was used to search for tumor-associated antigens by panning with a lung adenocarcinoma cell line, 1264, and counter-selecting with a nontumor bronchial epithelial cell line, BEAS-2B. After three rounds of subtractive panning, 239 of 673 clones analyzed bound selectively to 1264 tumor cells in a phage ELISA. Diversity analysis of these tumor-selective clones by BstNI fingerprinting and nucleotide sequencing revealed 14 distinct scFv fragments. Four clones bound selectively to 1264 over BEAS-2B cells when analyzed by a more discriminating flow cytometric assay using scFv. Moreover, these clones showed only limited cross-reactivity to several primary human cell lines. One clone, LU30, also cross-reacted strongly with the lung adenocarcinoma line, A549. The LU30 antigen was identified as decay-accelerating factor (CD55) by expression cloning from a 1264 cDNA library. The mean number of decay-accelerating factor molecules on the surface of 1264 and BEAS cells used for panning and counter-selection was estimated as 75,000 +/- 5,000 and 13,000 +/- 10,000, respectively. Thus, phage library panning combined with expression cloning permits identification of antibodies and their cognate antigens for proteins that are differentially expressed on the surface of distinct cell populations.     

Murine dendritic cells transduced with an adenoviral vector expressing a defined tumor antigen can overcome anti-adenovirus neutralizing immunity and induce effective tumor regression

In this study transduced dendritic cells (DCs) were used to enhance immunogenecity of a specific tumor antigen. Using a polyoma middle T (PyMT) transgenic mammary carcinoma model we found that injections of DCs transduced with an adenoviral (Ad) vector expressing PyMT (DCAd-PymT) led to potent specific anti-tumor immunity. Efficacy was not affected by neutralizing Abs (high or low titers) and naive animals did not produce detectable anti-Ad Abs following two injections of transduced DCs. Repeated injections of transduced DCs significantly improved therapeutic efficacy in mice with established lung metastases. These data emphasize the ability of Ad-infected DCs to: i) minimize anti-Ad Ab production, ii) overcome pre-existing anti-Ad humoral immunity, and iii) improve vaccination efficacy when injected more than once.