bluuebunny/crossref_metadata_2025_split huggingface dataset:
Macroprolactinemia in Childhood and Adolescence: A Cause of Asymptomatic Hyperprolactinemia
Authors: Fideleff H.L. , Ruibal G. , Boquete H. , Pujol A. , Sequera A. , Sobrado P. 
Published: April 2003

Asymptomatic hyperprolactinemias associated with altered proportions of molecular forms of circulating prolactin (PRL) have been reported in adults. The scarce references available in children and adolescents prompted us to report our experience in the evaluation and follow-up of patients with macroprolactinemia. We studied 5 patients (1 male and 4 females) aged 11.6–18 years with incidentally discovered asymptomatic hyperprolactinemia. Patients underwent repeated evaluations for a period of 3 months to 8 years, and their PRL levels remained elevated (34.4–516 ng/ml). Structural variants of PRL ≥45 kD ranged between 58.9 and 78.6%. Chromatographic profiles showed increases in Big Big PRL in the 5 cases, ranging between 40 and 72% (normal: 9–21%), and in Big PRL in 3 cases, ranging between 30.0 and 32.6% (normal: 5–25%). Little PRL was decreased in all cases, ranging between 20.6 and 41.1% (normal: 50– 90%). In conclusion, upon detection of hyperprolactinemia with no clinical manifestations and no alteration of the remaining endocrine functions, macroprolactinemia should be considered as a possible diagnosis. The confirmed absence of functional alterations during the follow-up would favor a no-treatment approach and at the same time avoid repeating imaging studies.
Pituitary Surgery for the Management of Acromegaly
Authors: Laws Edward R. , Vance Mary Lee , Thapar Kamal 
Published: April 2003

Active acromegaly is almost always the result of a benign growth hormone (GH)-secreting adenoma of the pituitary gland. Because the same pituitary stem cell can produce both GH and prolactin (PRL), many acromegalic patients also have hyperprolactinemia. The advantages of surgical excision of pituitary adenomas associated with acromegaly include: (1) prompt decrease in GH; (2) reliable and immediate relief of the mass effect from the tumor (decompression of the optic nerves and chiasm), and (3) the opportunity to obtain tumor tissue for characterization and investigative study. Currently, more than 97% of operations for removal of pituitary tumors associated with acromegaly are done using the transsphenoidal approach rather than craniotomy. Technical advances to make the surgery safer continue to evolve, and include endoscopic approaches, computer-guided image-based intraoperative visualization, and intraoperative magnetic resonance imaging. Criteria for satisfactory remission of acromegaly after surgery are the same as those used for medical management. They include normal insulin-like growth factor (IGF)-I and suppression of GH to undetectable levels (<1.0 ng/ml) during an oral glucose tolerance test (OGTT). Data from a recent series of 86 patients operated upon for acromegaly at the University of Virginia and followed for more than 1 year have been reviewed. In patients receiving surgery as the initial procedure, 67% had a normal IGF-I, and 52% suppressed to <1.0 ng/ml in an OGTT. There was one true recurrence of disease diagnosed 81 months after surgery. Results are best in patients with noninvasive microadenomas. Gamma knife radiosurgery has been a valuable adjunct in those patients who fail to achieve postoperative remission. Pathological evaluation of the tumors revealed that 16% expressed GH only, 25% stained for GH and glycoprotein hormones (follicle stimulating hormone, thyroid hormone, thyroid stimulating hormone, α-subunit), 21% for GH and PRL, and 33% for GH, PRL and glycoprotein hormones. There was one acidophil stem cell tumor and 10% had the mammosomatotroph subtype. This contemporary series was free of mortality or serious complications. One patient had a transient cerebrospinal fluid leak and 3 developed transient SIADH with hyponatremia. Surgical treatment remains an important aspect of the combined management of patients with acromegaly.
An Amplified ELISA Inhibition Method for the Measurement of Airborne Soybean Allergens
Authors: Cruz Maria-Jesús , Rodrigo Maria-José , Antó Josep-Maria , Morell Ferran 
Published: April 2003

<i>Background:</i> Measurement of the soybean aeroallergen in Barcelona and other cities where soybean is unloaded is of increasing importance in controlling population exposure and evaluating the influence of such exposure on the persistence of asthma symtpoms. <i>Objective:</i> The aims of the study were: (1) to standardize an amplified ELISA inhibition method for the quantification of soybean aeroallergen and (2) to compare this method to a previously described RAST inhibition method. <i>Methods and Results:</i> An amplified competitive ELISA inhibition method with a biotin-streptavidin system was carried out using a pool of sera from soybean-sensitized patients. The results were expressed as U/ml using a low-molecular-mass soybean allergen as reference standard. Reproducibility was calculated by statistically comparing the slope of the regression lines of the standard curve of 4 consecutive assays and by determining the coefficient of variation (CV) of the percent inhibition data for each point of several independent standard curves, each from the same assay (intra-assay) and also from a separate assay (inter-assay). No significant differences in the slopes were obtained by analysis of covariance (ANCOVA) F = 1.04. The CV between assays varied between 4 and 22% (for the assay range used in the reference standard) and was greater than the CV within assays (5–10%). Only values with a CV(%) smaller than 20% were considered acceptable. 78.5% of the samples satisfied this criterion. The RAST inhibition and ELISA inhibition methods were compared by difference plots from the values of 338 air filter eluates. The intraclass correlation coefficient was 0.456 (p < 0.001). After the results of both methods were classified as lower and higher than 165 U/m<sup>3</sup>, the kappa index was 0.46 (p < 0.001). <i>Conclusions:</i> The data obtained in the present study are comparable to those reported from other similar immunoassays. Moreover, despite the difficulty in comparing air-sampling values from different laboratories, the kappa index may be taken to represent fairly good agreement beyond chance between both methods. All these data demonstrate that the present immunoassay is useful for measuring airborne soybean aeroallergens and can also be applied to evaluate the relationship between exposure and the development of asthma symptoms.
Immunological Activity of Recombinant Ole e 1 in Patients with <i>Olea europaea</i> Pollinosis
Authors: Quiralte J. , González E. , Arias de Saavedra J.M. , Villalba M. , Florido J.F. , Sáenz de San Pedro B. , Rodríguez R. 
Published: April 2003

<i>Background:</i> Recombinant allergens have potential advantages over conventional allergenic extracts. However, these recombinant allergens should be evaluated for their antigenic activity and compared with their natural counterparts before being used for clinical purposes. <i>Methods:</i> We studied 33 patients with seasonal rhinitis and/or bronchial asthma and a positive skin prick test to <i>Olea europaea</i> pollen extract, 10 atopic patients with no history of pollinosis and a negative skin prick test to <i>O. europaea</i> extract and 10 healthy controls. Skin prick tests and determination by ELISA of specific IgE to natural Ole e 1 (nOle e 1) and recombinant Ole e 1 (rOle e 1) expressed in <i>Pichia pastoris</i> were performed in all patients and controls. Inhibition assays were performed between nOle e 1 and rOle e 1 by ELISA. <i>Results:</i> All patients with <i>O. europaea</i> pollinosis had positive skin test responses to both commercial <i>O. europaea</i> extract and nOle e 1 allergen, and all reacted to rOle e 1 on the skin prick test. The nonatopic and atopic control subjects with negative olive pollen skin test results did not react to rOle e 1 on the skin prick test, even at the highest concentrations, confirming the specificity of this test. We found a weak correlation between the wheal surface area produced by the prick test with nOle e 1 and the wheal surface area produced by rOle e 1 at 10 μg/ml (r = 0.42, p < 0.05). Comparison of specific IgE against both nOle e 1 and rOle e 1 in the patients did not reveal any significant difference. There was a strong correlation between the amount of specific IgE against nOle e 1 and rOle e 1 (r = 0.99, p < 0.01). The two proteins displayed the same extent of binding inhibition to IgE antibodies in ELISA inhibition experiments. <i>Conclusions:</i> These results confirm the immunological activity of rOle e 1 expressed in <i>P. pastoris</i> and indicate that Ole e 1 is one of the major allergens in <i>O. europaea</i> pollinosis as evaluated by skin prick test and serological methods. The correlation between rOle e 1 and nOle e 1 in skin test results and serologic data indicates the potential of recombinant allergens for clinical applications and diagnosis of <i>O. europaea</i> pollen allergy.
Production of Enzymatically and Immunologically Active Der f 1 in <i>Escherichia coli</i>
Authors: Takahashi Kyoko , Takai Toshiro , Yasuhara Takaomi , Yuuki Toshifumi , Ohtake Yasuyuki , Yokota Toyokazu , Okumura Yasushi 
Published: April 2003

Der f 1 is a major house dust mite allergen belonging to the cysteine protease family. Because of the great demand for clinical and research use of this allergen, much effort to establish an efficient method of preparing purified Der f 1 has been made. We constructed an isopropyl-β-<i>D</i>-thiogalactopyranoside-inducible expression plasmid to produce the pro-form of Der f 1 in <i>Escherichia coli.</i> The recombinant product was accumulated as insoluble inclusion bodies in cells. The solubilized inclusion bodies were found to be successfully renatured by two-step gel filtration chromatography. About 70 mg of pro Der f 1 were properly refolded by this method from 1 liter of culture. Acid treatment of the renatured pro Der f 1 resulted in the autocatalytic removal of the pro-sequence. The obtained mature form of Der f 1 bound IgE in patient sera and induced the release of histamine from peripheral blood leukocytes equally to native Der f 1. Furthermore, mature Der f 1 obtained by this method had identical protease activity with native Der f 1. We also discussed the contribution of the pro-sequence and the sugar chain of Der f 1 to its antigenic and enzymatic activity. This is the first report to produce an active mature form of recombinant Der f 1 in <i>E. coli.</i>
Elaboration of Matrix Metalloproteinase Inhibitors by Human Skin in Organ Culture and by Skin Cells in Monolayer Culture: Relationship to Invasion
Authors: Chi Yiqing , Zeigler Mary E. , Walker James , Perone Patricia , Datta Subhash C. , Varani James 
Published: April 2003

Functional and immunochemical approaches were used to assess matrix metalloproteinase (MMP) inhibitors, e.g., tissue inhibitor of metalloproteinases 1 and 2 (TIMP-1 and TIMP-2), in organ cultures of normal human skin maintained under growth factor free conditions or in medium supplemented with a combination of growth factors including epidermal growth factor, insulin, and pituitary extract. It has previously been shown that under growth factor free conditions, normal skin structure and function are maintained for several days, while in the presence of these exogenous growth factors, the epithelial cells invade the stroma [Invasion and Metastasis 1993;13:225–233]. TIMP-1 was detected in equivalent amounts in organ culture fluids under both conditions. TIMP-2 was not detected under either condition. Normal epidermal keratinocytes, normal dermal fibroblasts, and three different epithelial tumor cell lines were also examined for MMP inhibitor expression. Keratinocytes and fibroblasts produced high levels of both TIMP-1 and TIMP-2, but in neither cell type was there a significant difference between growth factor free and growth factor containing conditions. In contrast, the three epithelial tumor cell lines produced low to undetectable levels of both TIMP-1 and TIMP-2. These data suggest that acquisition of local invasive capacity is not dependent on a reduction in MMP inhibitor expression. A reduction in MMP inhibitors may accompany the transition from invasive to metastatic tumors.
Effects of All-<i>trans</i>-Retinoic Acid Incorporated into Low-Density Lipoprotein on Invasive Properties of Multidrug-Resistant MCF-7 Spheroids
Authors: Affoué M. , Akeli M.G. , Gueddari N. , Jardillier J.C. , Madoulet C. 
Published: April 2003

Cultured cells grown as spheroids provide an in vitro model that is closer to an in vivo tumour than conventional monolayer techniques. Previous work from our laboratory has demonstrated that spheroids formed from multidrug-resistant MCF-7 cells exhibit invasive characteristics which were not present in their sensitive counterparts. The treatment of these spheroids by all-<i>trans</i>-retinoic acid (ATRA), a potent inducer of in vitro and in vivo differentiation, decreases their proteolytic activity and ability to invade Matrigel-coated filters. The efficiency of ATRA is enhanced by its incorporation into low-density lipoprotein (LDL) (LDL-ATRA). Indeed, invasion through a reconstituted basement membrane was reduced by 73% with 10<sup>–6</sup> <i>M</i> ATRA and 3 × 10<sup>–8</sup> <i>M</i> LDL-ATRA. Furthermore, inhibition of invasion was correlated with a decrease in several factors: (1) secreted matrix metalloproteinase-9 and enzymes degrading type IV collagen and Matrigel films, and (2) tissue plasminogen activator. The results observed were found with a concentration of LDL-ATRA 30 times lower than that of ATRA. This could be due to the protective effect of LDL and to a better targeting of cancer cells through their LDL receptors. LDL-ATRA may therefore represent a new and potent inhibitor of invasion that could be developed for clinical trials.
Neutralization of Borna Disease Virus Depends upon Terminal Carbohydrate Residues (α-<i>D</i>-Man, β-<i>D</i>-GlcNAc) of Glycoproteins gp17 and gp94
Authors: Stoyloff Roman , Bode Liv , Borchers Kerstin , Ludwig Hanns 
Published: April 2003

Borna disease virus (BDV) is an enveloped, nonsegmented, negative-stranded RNA virus that causes infections of the brain in a wide range of animal species and man. The third open reading frame codes for a protein of 17 kD (gp17) that is N-glycosylated and contains terminal α-<i>D</i>-mannose and N-acetyl-β-<i>D</i>-glucosamine residues. Rat sera raised against these carbohydrates (anti-sugar antisera) show high in vitro neutralization activity and were capable of precipitating BDV. The neutralizing capacity of sera derived from experimentally BDV-infected rabbits, in turn, decreased after adsorption with those carbohydrates. They partially inhibited infection of primary young rabbit brain cells in a dose-dependent manner. Furthermore, the anti-sugar antisera recognized a second virus-specific glycoprotein with an apparent molecular mass of 94 kD (gp94), providing indirect evidence that gp94 is involved in virus adsorption and/or entry into cells. Neutralization of BDV comprises a complex event and, as shown for the first time, involves the carbohydrate residues of both glycoproteins of BDV.
A Novel Protein Tag from Herpes Simplex Virus Type 1 DNA Polymerase
Authors: Schreiner Uwe , Hansen Jutta , Kühn Frank J.P. , Knopf Charles W. 
Published: April 2003

An epitope (HPOL) derived from the so-called thumb region of the herpes simplex virus type 1 DNA polymerase in combination with a monoclonal antibody (MAb 1051c) was tested for protein tagging. Using a conventional expression vector, a DNA cassette encoding the HPOL epitope was fused to the C-terminus of the dihydrofolate reductase (DHFR) gene such that the recombinant DHFR contained both a N-terminal HIS-tag and a C-terminal HPOL tag. Expression of recombinant DHFR in <i>Escherichia coli</i> cells was compared by Western blot analysis using either mouse RGS·HIS antibody or MAb 1051c. Immunostaining revealed that both antibodies reacted specifically with DHFR, but the detection sensitivity achieved with MAb 1051c was about 15-fold greater using a standard staining protocol. An HPOL antibody column was successfully applied for affinity purification of DHFR, demonstrating the usefulness of the HPOL epitope/MAb 1051c system for protein tagging, expression monitoring and purification of HPOL-tagged recombinant proteins.
Modulation of Arterial Growth of the Rabbit Carotid Artery Associated with Experimental Elevation of Blood Flow
Authors: Di Stefano Ida , Koopmans Donna R. , Langille B. Lowell 
Published: April 2003

We examined the growth of the right common carotid artery of young rabbits after ligating the left common carotid artery at 3 weeks of age, a procedure that approximately doubled right carotid blood flows. Flow increased from 0.065 ± 0.003 to 0.096 ± 0.009 ml/s within 1 h and, at 15 weeks of age, carotid blood flows in experimental animals (0.747 ± 0.102 ml/s) were more than double of those of sham-operated control animals (0.334 ± 0.053 ml/s). Contralateral carotid ligation resulted in more rapid increases in diameter of the artery with growth in the experimental animals. At 15 weeks of age, the vessel was 15% larger than that of sham-operated controls (2.70 ± 0.09 vs. 2.34 ± 0.05 mm). This more rapid growth of diameter resulted in shear stresses that were not different from controls despite the higher blood flow rates. Interestingly, however, shear stresses in control arteries fell from 17.4 ± 3.4 to 9.19 ± 1.16 dyn/cm<sup>2</sup> over the experimental period (p < 0.05). Elastin accumulation in the experimental artery was much more rapid than in controls and elastin contents were 49% more than in controls at 15 weeks of age. DNA and collagen contents were not significantly affected by contralateral carotid ligation. Previously, we found that experimental manipulations that decreased flow in the same artery of weanling rabbits substantially affected elastin and DNA accumulation, but had no effect on collagen contents. We conclude that increased blood flow is associated with arterial growth and specifically with accumulation of elastin, a wall constituent that bears much of the wall tension at resting blood pressure, and therefore is a primary determinant of resting vessel dimensions.
Regulation of Insulin-Like Growth Factor-1 Expression in Vascular Endothelial Cells by the Inflammatory Cytokine Interleukin-1
Authors: Glazebrook Heather , Hatch Timothy , Brindle Nicholas P.J. 
Published: April 2003

Insulin-like growth factor-1 (IGF-1) has important roles in vascular physiology and pathologies such as atherosclerosis. However, the factors that control expression of this growth factor in human vascular cells are largely unknown. In this study the effects of the inflammatory cytokine interleukin-1 (IL-1) on IGF-1 mRNA and peptide synthesis was examined in human endothelial cells. Cultured human umbilical vein endothelial (HUVE) cells were challenged with interleukin-1 and IGF-1 Ea and Eb mRNA levels were determined by nuclease protection assays and reverse transcription/polymerase chain reaction. IL-1 caused a dramatic increase in IGF-1 Ea mRNA in HUVE cells. Transcript levels peaked between 2 and 4 h of IL-1 treatment and declined over the subsequent 4 h. Consistent with its effect on mRNA, the inflammatory cytokine causes a 3-fold stimulation in production of IGF-1 peptide from endothelial cells. These results demonstrate increased IGF-1 mRNA and peptide in vascular endothelial cells stimulated with IL-1. This suggests that under conditions of local inflammation at the vessel wall the direct action of IL-1 on endothelium can lead to induction of IGF-1 which could promote the intimal hyperplasia often seen in these circumstances.
A Platelet-Activating Factor Antagonist (WEB 2170) Preserves Endothelium-Dependent Vasodilatation and Prevents Development of a Neo-Intima Induced by a Periarterial Collar in Rabbit Carotid Arteries
Authors: Yin Zhan-Li , Hickey Haruyo , Dusting Gregory J. 
Published: April 2003

Platelet-activating factor (PAF) may be involved in adhesion of leucocytes and migration of cells during vascular remodelling for it is expressed in leucocytes after cytokine priming and is required for cell adhesion. We studied the effects of WEB 2170, a potent PAF antagonist, on the development of an atheroma-like neo-intima induced by a peri-arterial collar in rabbits. Either WEB 2170 (3 mg/kg/day) or vehicle was given by subcutaneous injection once a day for 4 or 9 days, and on day 3 peri-arterial collars were applied to both carotid arteries in all animals. Two or 7 days after implanting the collars vasodilator responses to the endothelium-dependent vasodilator, acetylcholine and the endothelium-independent vasodilator, sodium nitroprusside were studied in isolated artery rings from both groups of rabbits. Neo-intima formation after 7 days (day 10 of treatment) was measured by light microscopy as the ratio of cross-sectional areas of intima and media, and expression of inducible nitric oxide synthase (iNOS) was studied by immunohistochemistry. PAF-induced platelet aggregation ex vivo was inhibited specifically in WEB 2170-treated rabbits. At day 5, acetylcholine-induced vasorelaxation in collared artery rings was markedly impaired as compared to control sections from both vehicle- and WEB 2170-treated rabbits. At day 10, acetylcholine-induced vasorelaxation in collared artery rings from vehicle rabbits was markedly less than in controls, but in WEB 2170-treated rabbits, the acetylcholine response in collared arteries was similar to control sections. Intimal thickening was much reduced in WEB 2170-treated rabbits, ratios of intima/media areas being vehicle: 0.21 ± 0.02 (n = 5) and WEB 2170: 0.07 ± 0.01 (n = 7; p < 0.01). Immunofluorescence showed expression of iNOS only in the neo-intima of vehicle-treated, collared arteries, but not in the residual neo-intima of WEB 2170-treated, collared arteries. These results suggest that WEB 2170 is effective in preserving endothelial function, prevents the development of neo-intima and blocks iNOS expression in the neo-intima in this model.
Involvement of L-Type Calcium Channels in Hypoxic Relaxation of Vascular Smooth Muscle
Authors: Herrera Gerald M. , Walker Benjimen R. 
Published: April 2003

In the systemic vasculature, hypoxia elicits a local vasodilator response that may be partially mediated by ionic channels on vascular smooth muscle, such as adenosine triphosphate sensitive K<sup>+</sup> channels. Recent electrophysiological studies suggest that hypoxia may also inhibit voltage-dependent Ca<sup>2+</sup> channels (L type) on peripheral vascular smooth muscle cells. We hypothesized that hypoxia elicits relaxation of vascular smooth muscle by inhibiting L-type Ca<sup>2+</sup> channels. In endothelium-denuded rat thoracic aortic rings contracted with phenylephrine, mild and moderate hypoxia (PO<sub>2</sub> 35 and 20 mm Hg, respectively) elicited significant relaxation. Pretreatment with the L-type Ca<sup>2+</sup> channel antagonist nifedipine completely inhibited mild hypoxic relaxation and diminished relaxation under moderate hypoxia, whereas glibenclamide, a blocker of adenosine triphosphate sensitive potassium channels, only attenuated the response to moderate hypoxia. In rings contracted with the L-type Ca<sup>2+</sup> channel agonist (–)BAY K 8644 both mild and moderate hypoxia elicited almost complete relaxation. Furthermore, in rings contracted with hyperkalemic solutions (85 m<i>M</i> K<sup>+</sup> or 120 m<i>M</i> K), mild and moderate hypoxia elicited significant relaxations. Thus, we conclude that hypoxia acts directly on vascular smooth muscle to cause relaxation in part by inhibiting L-type Ca<sup>2+</sup> channels.