bluuebunny/crossref_metadata_2025_split huggingface dataset:
Characterization of Permanent Cell Lines That Contain the AAV2 <i>rep-cap</i> Genes on an Epstein-Barr-Virus-Based Episomal Plasmid
Authors: Neyns Bart , Vermeij Joanna , Teugels Eric , De Rijcke Martine , Hermonat Paul , De Grève Jacques 
Published: April 2003

Recombinant adeno-associated virus (rAAV) has emerged as a promising gene therapy vector. Its development, however, has been hampered by the lack of a readily available efficient production method. We investigated the possibility of establishing permanent cell lines for the production of rAAV with a new Epstein-Barr-virus (EBV)-based episomal AAV <i>rep-cap</i> plasmid (pCEP-<i>rep/cap</i>). HeLa and 293 cells were stably transfected with plasmids that carry the AAV2 <i>rep/cap</i> genes under transcriptional control of their endogenous promoters (p5, p19 and p40) either on the pCEP-<i>rep/cap</i> or an integrated (pIM45) plasmid. For the ease of monitoring transgene expression in live cells, a rAAV vector expressing <i>gfp</i> (the green fluorescent protein gene, rAAV-<i>gfp/neo</i>) was used. Establishment of stable transfected cell lines with these plasmids proved feasible but their usefulness was limited because of their instability. Within 8–12 weeks after their establishment, stably transfected <i>rep-cap</i> cell lines invariably lost their function. In addition, the rAAV-<i>gfp/neo</i> vector we used was susceptible to mutation in stably transfected HeLa cells. Our observations demonstrate specific problems both at the level of <i>rep/cap</i> gene function and the rAAV genome that can occur with the establishment of rAAV production cell lines. These experiments should aid the further development of efficient rAAV production protocols.
Relationship between Leuko-Araiosis and Blood Pressure Variability in the Elderly
Authors: Puisieux François , Monaca Pascal , Deplanque Dominique , Delmaire Christine , di Pompeo Christophe , Monaca Christelle , Leys Didier , Pruvo Jean-Pierre , Dewailly Philippe 
Published: April 2003

Although leuko-araiosis is a common finding on computed tomographic (CT) scans of the brain, its pathogenesis remains uncertain. To investigate the association between blood pressure (BP) disturbances and leuko-araiosis, we retrospectively reviewed CT scans and 24-hour ambulatory blood pressure monitorings of 79 elderly patients (57 women and 22 men; mean age: 83.3 ± 6.4 years). Of the 79 patients, 50 were demented (30 had Alzheimer’s disease and 18 vascular dementia) and 29 were not demented. The leuko-araiosis score (LA score) was determined by using Rezek’s scale. To evaluate short-term variation of BP, we determined (1) the variability of systolic and diastolic BPs (SBP, DBP; within-subject standard deviation of all readings over a 24-hour period), (2) the coefficient of variability (variability of BP/mean BP) and (3) the maximal variation of BP (difference between the maximum and minimum 24-hour BPs). Higher LA scores were associated with higher SBPs in 24-hour, diurnal and nocturnal periods, higher maximal variation of SBP, greater variability of SBP during 24-hour, diurnal and nocturnal periods and greater coefficient of variability of SBP during sleep. Our study suggests that elevations and short-term variations of SBP may contribute to the pathogenesis of white matter lesions in elderly persons.
Opportunities for Public Health Genetics Trainees: Results of an Employer/Workplace Survey
Authors: Austin Melissa A. , Arnett Donna , Beaty Terri , Durfy Sharon , Fineman Robert , Gettig Elizabeth , Lochner Doyle Debra , Peyser Patricia , Sorenson James , Thompson John D. , Watts Carolyn 
Published: April 2003

<i>Objective:</i> To conduct the first employer/workplace survey identifying employment opportunities for graduates of programs with training in public health genetics in the USA, and to determine whether employment opportunities will increase in coming years. <i>Methods:</i> Six public health genetics training competencies were developed. A survey about workplace and employment opportunities was then conducted with mailings to (1) departments in schools of public health and departments of preventive medicine, (2) local and regional public health officials, (3) insurance companies and health management organizations (HMOs), and (4) biotechnology and pharmaceutical companies. <i>Results:</i> A total of 196 surveys were returned among 1,464 that were mailed. Response rates varied from 5.8 to 46.5% among the target groups. The percent of responding organizations currently employing individuals with skills in genetics ranged from 20 to 62%. The percent currently employing individuals with skills in public health ranged from 39 to 96%. Training opportunities such as internships or practicum experiences are reported for one-third of respondents. For all of the competencies, approximately half of survey respondents who rated the competency important or very important already employ individuals with public health genetics skills. Similarly, at least a quarter of survey respondents who rated the competency important or very important plan to hire individuals with that skill in the next 5 years. Overall, approximately 40% of those surveyed are planning to hire individuals with competencies in public health genetics in the next 5 years. <i>Conclusion:</i> Employment opportunities already exist and new positions are becoming available in schools of public health and departments of preventive medicine, departments of public health, insurance companies and HMOs for professionals with public health genetics training. Based on our survey findings, skills and training in public health genetics are important in the workplace.
High-Dose Pulse Corticosteroid Therapy in the Treatment of Severe Alopecia areata
Authors: Seiter Simone , Ugurel Selma , Tilgen Wolfgang , Reinhold Uwe 
Published: April 2003

<i>Objective:</i> The present monocenter prospective study was designed to evaluate the efficacy of intravenous high-dose methylprednisolone pulse therapy in patients with severe alopecia areata (AA). <i>Methods:</i> 30 patients (aged 14–56 years) were treated with methylprednisolone (8 mg/kg body weight) intravenously on 3 consecutive days at 4-week intervals for at least 3 courses. <i>Results:</i> 67% of patients with AA plurifocalis showed >50% regrowth of hair. None of the patients with AA totalis or universalis and only 1 patient with ophiasic AA responded to therapy. In patients with AA plurifocalis, higher response rates could be observed in those suffering from long-term disease compared to patients treated during their first episodes of AA (73 vs. 57%). <i>Conclusion:</i> High-dose methylprednisolone pulse therapy is an effective and well-tolerated treatment for patients with severe AA plurifocalis but might be less beneficial for patients with ophiasic AA, AA totalis or universalis.
Topical Metronidazole Does Not Abate UVB-Induced Erythema
Authors: Guarrera Marcella , Brusati Carlo , Rebora Alfredo 
Published: April 2003

<i>Background:</i> The capacity of topical metronidazole to reduce erythema severity scores in rosacea has been attributed to its anti-inflammatory activity. <i>Methods:</i> Nine healthy subjects were irradiated on three 1-cm<sup>2</sup> areas on their volar forearm with 2 MEDs of UVB. Twenty-four hours later, one of the irradiated areas was treated daily with metronidazole 1% vanishing cream, one with the vehicle only and the third was left untreated (control). The intensity of erythema was measured, at 24, 48, 72 and 96 h after irradiation, by a Chromameter CR200 (Minolta, Japan) and expressed in the L*a*b* system. The a* parameter was taken as the measure of redness. <i>Results:</i> Overall, redness diminished over time in a linear way, but treatments did not influence it significantly. <i>Conclusion:</i> In our vehicle, metronidazole 1% cream failed to abate the UVB-induced erythema. An activity other than the anti-inflammatory one should be suggested to explain the topical metronidazole efficacy in rosacea.
Correlation between Respiratory Function and Airway Inflammation in Asthma
Authors: Hashimoto Tadanori , Minoguchi Kenji , Kohno Yasurou , Wada Kiyoko , Ueno Kana , Kobayashi Hideko , Miyamoto Masahide , Adachi Mitsuru 
Published: April 2003

<i>Background: </i>Eosinophilic airway inflammation is thought to be associated with airflow limitation and airway hyperresponsiveness. <i>Methods:</i> Spirometry, peak expiratory flow (PEF) measurement, histamine challenge test, and sputum induction with hypertonic saline inhalation were performed in 70 asymptomatic patients with asthma who were treated with bronchodilators or an inhaled corticosteroid (beclomethasone dipropionate, 400–800 µg/day) or both for more than 2 months. The relationships between the sputum eosinophils, respiratory function, and airway responsiveness to histamine were investigated. <i>Results:</i> Induced sputum was obtained from 53 patients. Although significant correlations between sputum eosinophils and forced expiratory volume in 1 s/forced vital capacity × 100% (FEV<sub>1</sub>%) or maximum expiratory flow at 25% of the forced vital capacity (&#118;&#775;<sub>25</sub>) divided by height (&#118;&#775;<sub>25</sub>/Ht) were observed, no significant correlations were found between sputum eosinophils and percent predicted PEF (%PEF) or airway responsiveness to histamine. Furthermore, airway responsiveness to histamine was not correlated with FEV<sub>1</sub>%, &#118;&#775;<sub>25</sub>/Ht or %PEF. <i>Conclusion:</i> Single-point measurements of FEV<sub>1</sub>% and &#118;&#775;<sub>25</sub>/Ht but not PEF or airway responsiveness to histamine correlate with the degree of airway inflammation evaluated indirectly with induced sputum.
Suppression of ICAM-1 Expression in Renal Proximal Tubular Cells by 1,25-Dihydroxyvitamin D<sub>3</sub>
Authors: Weinreich Thomas , Wüthrich Rudolf P. , Booy Christa , Binswanger Ulrich 
Published: April 2003

<i>Background:</i> 1,25-Dihydroxyvitamin D<sub>3</sub> is mainly synthesized by renal proximal tubular cells. More recently, it has been shown to affect cell growth and TGF-β<sub>1</sub> synthesis in glomerular and tubular renal cells in vitro, and to prevent glomerulosclerosis in vivo in subtotally nephrectomized rats. The mechanisms involved have not been fully identified. We asked whether 1,25-vitamin D<sub>3</sub> might interact with additional immunoregulatory functions of renal cells by studying its effects on the expression of the cellular adhesion molecules ICAM-1 (CD54) and VCAM-1 (CD106) in human proximal tubular cells in vitro (HK-2 cells). <i>Methods:</i> Expression of adhesion molecules was assessed in HK-2 cells cultured under basal conditions and after stimulation with TNF-α plus IFN-γ, by flow cytometry, gene transcription (RT-PCR) and measurement of soluble ICAM-1 in culture supernatant by ELISA. <i>Results:</i> Unstimulated HK-2 cells did not express VCAM-1 and only little ICAM-1. 1,25-Vitamin D<sub>3</sub> had no effect on the expression of adhesion molecules in unstimulated cells. TNF/IFN stimulation resulted in a 4-fold increase in ICAM-1 and VCAM-1 expression. The TNF/IFN-induced increase in ICAM-1 expression was reduced by 1,25-vitamin D<sub>3</sub> dose dependently (10<sup>–7</sup> <i>M</i> vs. solvent: –30%; 10<sup>–9</sup> <i>M:</i> –18%; 10<sup>–11</sup> <i>M:</i> –17%). 25(OH)-vitamin D<sub>3</sub> had no effect. ICAM-1 mRNA concentration was increased in TNF/IFN-stimulated cells. 1,25-Vitamin D<sub>3</sub> treatment prevented the increase of ICAM-1 mRNA by 27% after 24–72 h incubation (p = 0.03). The TNF/IFN-induced increase in soluble ICAM in culture supernatants was unchanged by 1,25-vitamin D<sub>3</sub>. VCAM-1 expression was unchanged by incubation with 1,25-vitamin D<sub>3</sub> under basal conditions and after TNF/IFN stimulation. <i>Conclusion:</i> 1,25-Vitamin D<sub>3</sub> inhibits cytokine-induced ICAM-1 but not VCAM-1 expression in renal proximal tubular cells in vitro. The present data support the hypothesis that 1,25-vitamin D<sub>3</sub> is not only synthesized by renal tubular cells, but may also affect immunoregulatory functions in these cells.
Intrarenal Distribution of Blood Flow in Sodium Depleted and Sodium Loaded Rats: Role of Nitric Oxide
Authors: Hably Csilla , Vág János , Tost Hilda , Csabai Zsuzsanna , Bartha Jenő 
Published: April 2003

The renal hemodynamic effects of nitric oxide synthase (NOS) inhibition and dietary salt were studied in rats. L-NAME (0.1 mg/ml in the drinking fluid, about 12 mg/kg/day) was given for 4 days to rats receiving low (sodium depletion, SD), normal (N) or high (sodium load, SL) NaCl diet. Intrarenal hemodynamics was studied in anaesthesia. NOS inhibition decreased renal blood flow and increased renal vascular resistance in each group. Cortical and outer medullary but not inner medullary blood flow increased in direct ratio to the sodium intake. NOS inhibition decreased the blood flow and increased the vascular resistance in all layers of the kidney in SD, N, and SL rats as well. In SD and N, but not in SL rats L-NAME induced vasoconstriction was higher in the outer (OM) and inner medulla (IM) than in the cortex (C) [SD: ΔCVR 43%, ΔOMVR 54%, ΔIMVR 84%; N: ΔCVR 54%, ΔOMVR 96%, ΔIMVR 106%; SL: ΔCVR 50%, ΔOMVR 64%, ΔIMVR 35%]; in normal rats blood flow shifts from the medulla toward the cortex. In conclusion, nitric oxide may have a role in the regulation of renal vascular tone not only in the case of regular sodium uptake but in the sodium depleted or loaded organism as well. However, nitric oxide has no role in the dietary salt evoked vascular adaptation in the kidney.
Melatonin Modulation of the Daily Prolactin Secretion in Intact and Ovariectomized Ewes
Authors: Misztal Tomasz , Romanowicz Katarzyna , Barcikowski Bernard 
Published: April 2003

The aim of this study was to investigate whether melatonin might modulate the daily prolactin secretion in the ewe during a period of ovarian activity and, if so, whether this modulatory action of melatonin was related to the presence of estradiol in the organism. Ewes in the late follicular and luteal phase, as well as overiectomized ewes without (OVX) and after 7 days of estradiol injections (OVX+E<sub>2</sub>) were examined. Melatonin was infused into the third brain ventricle (100 µg/100 µl/h) from 14.00 to 18.00 h. The concentration of prolactin increased significantly during the infusion of melatonin in late follicular-phase ewes, but not in luteal-phase ewes, as compared to the concentration before the infusion: range from 204.0 ± 31.7 to 272.2 ± 50.1 ng/ml vs. range from 68.2 ± 31.8 to 94.7 ± 33.1 ng/ml (mean ± SEM, n = 4, p < 0.01) and to the concentration noted during control infusions: range from 130.0 ± 58.0 to 179.3 ± 55.6 ng/ml (mean ± SEM, n = 4, p < 0.05). In ovariectomized ewes, the concentration of prolactin during infusion of melatonin increased significantly, unrelated to the presence of estradiol, as compared to the concentration before infusion: range from 136.7 ± 20.3 to 260.0 ± 11.6 ng/ml vs. range from 41.6 ± 2.6 to 152.3 ± 14.6 ng/ml in OVX ewes (mean ± SEM, n = 4, p < 0.01) and range from 161.5 ± 66.5 to 250.2 ± 24.3 ng/ml vs. range from 61.2 ± 1.7 to 159.2 ± 43.3 ng/ml in OVX+E<sub>2</sub> ewes (mean ± SEM, n = 4, p < 0.01). Concentrations during infusion of melatonin in OVX and OVX+E<sub>2</sub> ewes were also significantly higher than during the control infusions: range from 7.2 ± 1.7 to 22.2 ± 4.1 ng/ml (mean ± SEM, n = 4, p < 0.001). These results indicate that melatonin may affect the daily secretion of prolactin in ewes during the breeding season, and suggest that the variable response of prolactin to the melatonin signal in intact and ovariectomized ewes relates to the interaction between both ovarian steroids – estradiol and progesterone – and the prolactin-releasing factor.
Effects of Leptin on Melanin-Concentrating Hormone Expression in the Brain of Lean and Obese Lep<sup>ob</sup>/Lep<sup>ob</sup> Mice
Authors: Huang Qingling , Viale Agnès , Picard Frédéric , Nahon Jean-Louis , Richard Denis 
Published: April 2003

The effect of leptin on the expression of melanin-concentrating hormone (MCH) was investigated in lean and genetically obese Lep<sup>ob</sup>/Lep<sup>ob</sup> mice. Murine leptin was subcutaneously infused using osmotic minipumps. The treatment period extended to 7 days and the daily dose of leptin delivered was 100 µg/kg of body weight. In situ hybridization was used to assess the effects of leptin infusion on the expression of MCH mRNA. MCH levels in the brain (hypothalamus/thalamus and the remaining part), spleen and testis were measured by radioimmunoassay coupled to HPLC of selected tissue extracts. Leptin significantly reduced final body weight, weight of brown adipose tissue, daily food intake in obese mice but not in lean animals. In obese mice, leptin led to a rapid reduction in food intake which reached statistical significance after only 24 h and which led to a significant reduction in the body weight after 3 days of treatment. Leptin restored the normal circulating levels of glucose and insulin in obese mice. The present results mainly provide evidence for a stimulating effect of leptin infusion on the MCH neuronal system. The hypothalamic/thalamic levels of MCH mRNA and peptide were higher in mice treated with leptin than in mice infused with phosphate-buffered saline. The stimulation of MCH neurons appeared particularly intense in obese mice, in which the effects of leptin infusion led to the reduction in MCH content of neuron fibers and terminals. Leptin did not affect spleen and testis MCH contents. In the light of the acknowledged orexigenic effects of MCH, the results of this study questioned the direct role of MCH in the action of leptin on energy balance. The increase in MCH expression following leptin could occur as a mechanism to compensate the decrease in energy deposition led to by leptin. It may also indicate that MCH mediates the metabolic actions of leptin indirectly or else that leptin influences actions of MCH other than those related to the regulation of energy balance.
Central Administration of Antiserum to Vasoactive Intestinal Peptide Delays and Reduces Luteinizing Hormone and Prolactin Surges in Ovariectomized, Estrogen-Treated Rats
Authors: vanderBeek ElineM. , Swarts HansJ.M. , Wiegant VictorM. 
Published: April 2003

The present study investigated the role of hypothalamic VIP in the regulation of the LH and PRL surge using immunoneutralization of endogenous VIP in mature ovariectomized (OVX), estradiol benzoate (EB)-treated female Wistar rats. We compared the effect of intracerebroventricular (i.c.v.) injections of a VIP antiserum (VIP-Ab) with that of saline (Ctr) on LH and PRL profiles in two separate groups of rats following two subcutaneous EB injections on days 8 and 9 after OVX. VIP-Ab or Ctr injections were given during the second half of the dark period, i.e. at 22:00 h (day 9), and, in addition, the following morning, i.e. at 08:00 h (day 10), just before the expected onset of the LH surge. Hourly blood samples were collected between 09:00 and 18:00 h on day 10. In addition, we studied the reproducibility of EB-induced LH and PRL surges and compared the effect of Ctr and VIP-Ab treatment on sequential surges in individual OVX females, i.e. 10 and 23 days after OVX, using each animal as its own control. Although we observeda large variation in the height and timing of LH and PRL peak levels between EB-treated females, the characteristics of successive surges of individual rats were highly reproducible. This reproducibility suggests that differences in functioning of the suprachiasmatic nucleus as well as in the response of the hypothalamus to steroid feedback largely explain the normal variation in hormone responses between rats. The VIP-Ab treatment resulted in a significant delay in the time course and a strong reduction of the magnitude of the afternoon LH and PRL surge. When analyzed within individual females, the effect of VIP-Ab treatment was even more pronounced due to a reduction in variability when each animal was used as its own control. These results suggest that hypothalamic VIP is an important regulator of both the timing and the magnitude of the EB-induced LH and PRL surge in the OVX rat, and suggest that its role may be stimulatory in this respect.
Noradrenergic and Dopaminergic Activity in the Hypothalamic Paraventricular Nucleus after Naloxone-Induced Morphine Withdrawal
Authors: Fuertes Gregorio , Laorden M. Luisa , Milanés M. Victoria 
Published: April 2003

Previous research has shown an increase in hypothalamo-pituitary-adrenal axis activity following naloxone administration to morphine-dependent rats. In the present study, we investigated the adaptive changes in the noradrenaline (NA) and dopamine (DA) systems in the hypothalamic paraventricular nucleus (PVN) during morphine dependence and withdrawal. Additionally, we examined the possible change in 3′,5′-cyclic adenosine monophosphate (cAMP) levels in that nucleus under the same conditions. Rats were made dependent on morphine by morphine or placebo (naïve) pellet implantation for 7 days. On day 8, rat groups received an acute injection of saline or naloxone (1 mg/kg subcutaneously) and were decapitated 30 min later. NA and DA content as well as their metabolite production in the PVN were estimated by HPLC/ED. Both plasma corticosterone levels and cAMP concentration in the PVN were measured by RIA. Naloxone administration to morphine-dependent rats (withdrawal) induced a pronounced increase in the production of both the NA metabolite MHPG and the DA metabolite DOPAC and an enhanced NA and DA turnover. Furthermore, an increase in corticosterone secretion was observed in parallel to the changes in catecholamine turnover. However, no alterations in cAMP levels were seen during morphine withdrawal. These results raise the possibility that catecholaminergic afferents to the PVN could play a significant role in the alterations of PVN functions and consequently in the pituitary-adrenal response during morphine abstinence syndrome. These data provide further support for the idea of adaptive changes in catecholaminergic neurons projecting to the PVN during chronic morphine exposure.
Analysis of the Metabolism of Haloperidol and Its Neurotoxic Pyridinium Metabolite in Patients with Drug-Induced Parkinsonism
Authors: Iwahashi K. , Anemo K. , Nakamura K. , Fukunishi I. , Igarashi K. 
Published: April 2003

The blood levels of the neurotrophic drug haloperidol (HP) and its pyridinium metabolite, HPP<sup>+</sup>, have been analyzed by liquid chromatography/electrospray ionization-mass spectrometry in 10 schizophrenic patients treated with HP, without carbamazepine (HP, oral daily dose of 0.3–0.5 mg/kg body weight for more than 1 year, females, aged 41 ± 8.5 years). There was a significant difference (t-test, d.f. = 8, p (t&#8320; = 7.2) <0.005) in the blood HPP<sup>+</sup> level between the 5 patients with (18.5 ± 6.4 ng/ml) and the 5 without (6.3 ± 2.4 ng/ml) severe side effects such as drug-induced parkinsonism (Extrapyramidal Symptom Rating Scale (ESRS) parkinsonism severity scores 2.8 ± 1.5 and 1.8 ± 1.1, respectively). Moreover, it is suggested that vitamin E may be effective for drug-induced parkinsonism through a change in the blood HPP<sup>+</sup> level. It is necessary to investigate the HPP<sup>+</sup> metabolism in psychiatric patients to avoid severe side effects such as drug-induced parkinsonism and cardiac functional disorders.