bluuebunny/crossref_metadata_2025_split huggingface dataset:
A global standard for anti‐D immunoglobulin: international collaborative study to evaluate a candidate preparation
Authors: Thorpe S. J. , Sands D. , Fox B. , Behr‐Gross M.‐E. , Schäffner G. , Yu M. W. 
Published: November 2003

Background and Objectives  The aim of the study was to evaluate a lyophilized anti‐D immunoglobulin preparation to serve as a global standard for potency assays of anti‐D immunoglobulin products.Materials and Methods  The candidate global standard, 01/572, was calibrated against the World Health Organization (WHO) International Reference Preparation (IRP) for anti‐D immunoglobulin, human (68/419), along with two reserve candidate reference preparations, in an international collaborative study involving 25 laboratories in 15 countries. The United States Food and Drug Administration (US‐FDA) Center for Biologics Evaluation and Research (CBER) Standard for anti‐D immunoglobulin, Lot 3, was included for comparison. Most laboratories (20/25) performed AutoAnalyser methodology, competitive enzyme‐linked immunoassay (EIA) and/or flow cytometry.Results  The overall mean potency of the candidate global standard, 01/572, was 284·5 international units (IU)/ampoule, with an interlaboratory variability, expressed as a percentage geometric coefficient of variation (% gcv), of 9·7. The mean potency of the US Standard was 859·4 IU/ml with an interlaboratory variability of 9·5% gcv, excluding an outlier. The mean potencies of the reserve preparations per ampoule/vial were 110·6 IU and 106·7 IU when calibrated against the IRP, and 112·2 IU and 106·6 IU when calibrated against 01/572, respectively, with interlaboratory % gcv values of 9·6–18·3 (excluding outliers).Conclusions  Preparation 01/572 proved more suitable for use as a global standard than the reserve candidate preparations and was established, with an assigned potency of 285 IU/ampoule, by the WHO as the 2nd International Standard for anti‐D immunoglobulin; by FDA‐CBER as the Standard for anti‐D immunoglobulin, Lot 4; and by the European Directorate for the Quality of Medicines (EDQM) as the 1st Biological Reference Preparation for anti‐D immunoglobulin.
Antigen dose, type of antigen‐presenting cell and time of differentiation contribute to the T helper 1/T helper 2 polarization of naive T cells
Authors: Rothoeft T. , Gonschorek A. , Bartz H. , Anhenn O. , Schauer U. 
Published: November 2003

SummaryAntigenic encounter by T cells induces immunological synapse formation and T‐cell activation. Using different concentrations of toxic shock syndrome toxin‐1 (TSST‐1) as stimulus, we examined the capacities of dendritic cells (DC) and macrophages (Mφ) to prime syngeneic naive T cells. DCs were, under all experimental settings, more efficient than Mφ at clustering T cells. Translocation of the T‐cell receptor (TCR) to the contact area was found to be induced by DCs, as well as by Mφ, in an antigen‐dependent manner, although Mφ were less efficient at inducing TCR translocation. Capping of protein kinase C θ (PKCθ) was also antigen dependent but induced exclusively by DCs. Likewise, DCs were found to be more potent inducers of interleukin‐2 (IL‐2) production and proliferation of naive T cells than Mφ. After 3 days of culture, DCs presenting 100 ng/ml TSST‐1 induced interferon‐γ (IFN‐γ)‐secreting cells, whereas Mφ did not. After 7 days of culture, DCs presenting 0·1 ng/ml TSST‐1, and Mφ presenting high (as well as low) doses of TSST‐1, induced IL‐4‐producing cells. We therefore provide evidence to show that antigen dose, type of antigen‐presenting cell and time of differentiation can contribute to T‐cell differentiation.
Human T‐cell responses to the RD1‐encoded protein TB27.4 (Rv3878) from Mycobacterium tuberculosis
Authors: Agger Else Marie , Brock Inger , Okkels Limei Meng , Arend Sandra M. , Aagaard Claus S. , Weldingh Karin N. , Andersen Peter 
Published: November 2003

SummaryIn recent years, there has been considerable focus on the discovery and characterization of proteins derived from Mycobacterium tuberculosis leading to the identification of a number of candidate antigens for use in vaccine development or for diagnostic purposes. Previous experiments have demonstrated an important immunological role for proteins encoded by the RD1 region, which is absent from all strains of bacillus Calmette–Guérin (BCG) but present in the genomes of virulent M. bovis and M. tuberculosis. Herein, we have studied human T‐cell responses to the antigen encoded by the putative open reading frame (rv3878) of the RD1 region. Immunoblot analysis revealed that rv3878 was expressed and the native protein was designated TB27.4. Immunological evaluations demonstrate that TB27.4 elicits a prominent immune response in human tuberculosis patients with a dominant region in the C‐terminal part of the molecule. In contrast, very limited responses were seen in M. bovis BCG‐vaccinated donors. This study therefore emphasizes the diagnostic potential of proteins encoded by the RD1 region.
Up‐regulation of C5a receptor expression and function on human monocyte derived dendritic cells by prostaglandin E2
Authors: Weinmann Oliver , Gutzmer Ralf , Zwirner Jörg , Wittmann Miriam , Langer Katja , Lisewski Margarete , Mommert Susanne , Kapp Alexander , Werfel Thomas 
Published: November 2003

SummaryThe expression of the C5a‐receptor (C5aR) on dendritic cells, its regulation and function have not been well established thus far. We show that the C5aR is expressed on human monocyte‐derived dendritic cells (DC) and can be down‐regulated by maturation stimuli such as tumour necrosis factor‐α (TNF‐α), lipopolysaccharide (LPS) or CD40L and by the T helper 1‐cytokine interferon‐γ (INF‐γ). Prostaglandin E2 (PGE2), a proinflammatory mediator supporting dendritic cell activation and necessary for adequate DC migration, leads to the up‐regulation of C5aR expression when incubated alone and prevents down‐regulation when given in combination with TNF‐α or LPS. Stimulation of C5aR on DC triggered F‐actin polymerization, indicating the chemotactic potential of DC elicited by C5a. C5a induced F‐actin polymerization was increased when C5aR was up‐regulated by PGE2. Stimulation of DC with C5a resulted in interleukin‐10 production which was significantly increased after C5aR up‐regulation with TNF‐α and PGE2. Therefore, up‐regulation of the C5aR on human DC alters their chemotactic and immunologic response to C5a.
Autoreactive responses to an environmental factor: 1. phthalate induces antibodies exhibiting anti‐DNA specificity
Authors: Lim So‐Yon , Ghosh Swapan K. 
Published: November 2003

SummaryEnvironmental factors have been implicated in the induction of autoimmune disorders. We report here that a common chemical, phthalate, used widely in synthetic polymers and cosmetics induces serum anti‐self DNA antibodies in BALB/c, NZB and autoimmune‐prone NZB/W F1 mice. The latter group experiences a high mortality, and significantly higher anti‐DNA antibody levels along with nephritis and other histopathologic changes in kidney. Comparison of amino acid sequences of an anti‐phthalate BALB/c B‐cell hybrid, 2C3 with the known database at the National Center for Biotechnology Information reveals a striking homology between the variable regions of 2C3‐Ig (γ1, κ) and an anti‐DNA antibody, BV04‐01 (γ2b,κ) isolated from the lupus‐prone NZB/W F1 mice. The homology is 98% for κ light chain and 70% for γ heavy chain. Like 2C3‐Ig, BV04‐01 also has specificity for d(pT)4. Furthermore, the light chains of both 2C3‐Ig and BV04‐01 are products of Vκ1 gene. To understand the nature of anti‐phthalate responses in general, hybridomas generated from phthalate–keyhole limpet haemocyanin‐primed BALB/c splenocytes were characterized. The study identifies cross‐reactive populations that strongly bind phthalate, DNA, or both. Of the 14 hybridomas evaluated, six express the same Vκ1 gene‐derived light chain as 2C3, and bind both phthalate and ds and ss‐DNA. They specifically recognize the oligonucleotides, d(pT)4, and d(pT)10. Additionally, when antisera raised against idiopeptides corresponding to 2C3‐Ig hypervariable regions are allowed to react with 2C3‐Ig, their binding is blocked specifically by both d(pT)4 and phthalate. This study clearly demonstrates that phthalate exposure leads to activation of a significant number of autoreactive B‐cells, with the consequence of a significant pathogenic progression in susceptible NZB/W F1 mice but not in non‐autoimmune‐prone BALB/c.
Monitoring enzymatic reactions in nanolitre wells
Authors: Young I. T. , Moerman R. , Van Den Doel L. R. , Iordanov V. , Kroon A. , Dietrich H. R. C. , Van Dedem G. W. K. , Bossche A. , Gray B. L. , Sarro L. , Verbeek P. W. , Van Vliet L. J. 
Published: November 2003

SummaryWe have developed a laboratory‐on‐a‐chip microarray system based on nanolitre‐capacity wells etched in silicon. We have devised methods for dispensing reagents as well as samples, for preventing evaporation, for embedding electronics in each well to measure fluid volume per well in real‐time, and for monitoring the fluorescence associated with the production or consumption of NADH in enzyme‐catalysed reactions. Such reactions can be found in the glycolytic pathway of yeast. We describe the design, construction and testing of our laboratory‐on‐a‐chip. We also describe the use of these chips to measure both fluorescence (such as that evidenced in NADH) as well as bioluminescence (such as evidenced in ATP assays). We show that our detection limit for NADH fluorescence is 5 µm with a microscope‐based system and 100 µm for an embedded photodiode system. The photodiode system also provides a detection limit of 2.4 µm for ATP/luciferase bioluminescence.
The molecular machines of DNA repair: scanning force microscopy analysis of their architecture
Authors: Janićijević A. , Ristic D. , Wyman C. 
Published: November 2003

SummaryThe application of scanning force microscope (SFM, also called atomic force microscope or AFM) imaging to study the architecture of proteins and their functional assemblies on DNA has provided new and exciting information on the mechanism of vital cellular processes. Rapid progress in molecular biology has resulted in the identification and isolation of proteins and protein complexes that function in specific DNA transactions. These proteins and protein complexes can now be analysed at the single molecule level, whereby the functional assemblies are often described as nanomachines. Understanding how they work requires understanding their structure and functional arrangement in three dimensions. The SFM is uniquely suited to provide three‐dimensional structural information on biomolecules at nanometre resolution. In this review we focus on recent applications of SFM to reveal detailed information on the architecture and mechanism of action of protein machinery involved in safeguarding genome stability through DNA repair processes.
Self‐assembly and recrystallization of bacterial S‐layer proteins at silicon supports imaged in real time by atomic force microscopy
Authors: Györvary E. S. , Stein O. , Pum D. , Sleytr U. B. 
Published: November 2003

SummaryThe self‐assembly of bacterial surface‐layer (S‐layer) proteins (SbpA of Bacillus sphaericus CCM 2177) at silicon supports (hydrophobic, non‐plasma‐treated and hydrophilic, O2 plasma‐treated silicon supports) was imaged in real time by atomic force microscopy (AFM). A closed mosaic layer consisting of small crystals (less than 200 nm in diameter) was formed at a hydrophobic silicon support, whereas a coherent crystalline lattice consisting of large domains (2–10 µm in size) was generated at O2 plasma‐treated, hydrophilic silicon wafers. The structure of the formed layers was a monolayer (9 nm in height) at the hydrophobic silicon and a bilayer (15 nm in height) at the hydrophilic silicon. In situ AFM measurements confirmed the importance of ionic bonds in the formation of crystalline SbpA layers at silicon supports. Rupture of the protein subunits with a metal chelator from the crystalline lattice of SbpA was visualized in situ by AFM. The stability of solid‐supported SbpA layers could be enhanced by cross‐linking the S‐layers with amino–amino or amino–carboxyl group directed cross‐linkers.
The effect of seminal vesicle‐sparing radical prostatectomy on serum prostate‐specific antigen level
Authors: John H. , Hauri D. , Maake C. 
Published: November 2003

OBJECTIVETo investigate if the remaining seminal vesicle tips can affect serum levels of prostate‐specific antigen (PSA) in patients after seminal vesicle‐sparing radical prostatectomy (SVRP).PATIENTS AND METHODSThirty‐six patients were treated by either radical retropubic prostatovesiculectomy (23) or SVRP (13). Serum PSA was monitored in all patients before surgery, and at 6 weeks and 30 months afterward. Samples of normal seminal vesicles from radical cystectomies (six) were also snap‐frozen and either processed for semiquantitative reverse transcriptase‐polymerase chain reaction (RT‐PCR) using primers against PSA and α‐actin (for normalization) or for PSA immunohistochemistry.RESULTSRT‐PCR and sequencing showed that the seminal vesicles synthesise PSA mRNA. Furthermore, PSA peptide was detectable in the glandular epithelium of the seminal vesicle using immunohistochemical methods. There was no significant difference in serum PSA levels after standard or SVRP, with median (range) values (ng/mL) at 6 weeks of 0.04 (0.04–0.9) and 0.04 (0.04–0.66) and at 30 months of 0.17 (0.04–3.8) and 0.22 (0.04–58.2), respectively.CONCLUSIONAlthough the seminal vesicles produce PSA, the PSA derived from the remaining seminal vesicle tips after SVRP has no effect on the oncological follow‐up of these patients.
Retinal vascular findings and penile cavernosal artery blood flow
Authors: Kawanishi Y. , Kimura K. , Nakanishi R. , Numata A. , Taguchi H. 
Published: November 2003

Authors from Japan attempt to correlate retinal vascular changes with cavernosal arterial blood flow. They studied patients with erectile dysfunction, but excluded patients with previous pelvic surgery, pelvic injury or diabetes. They found that penile changes can be anticipated from retinal vascular changes seen on fundoscopy.OBJECTIVETo assess the correlation between retinal vascular findings and penile cavernosal arterial blood flow, as it is probable that systemic atherosclerotic vascular disease is important in male erectile dysfunction (ED), and being systemic, it might be possible to evaluate the extent of atherosclerosis from retinal vascular findings.PATIENTS AND METHODSThe study included 75 patients with ED; any with a history of pelvic injury, pelvic surgery, or diabetes mellitus were excluded. All patients gave fully informed consent. Ocular fundus photographs were taken with an automatic‐focus fundus camera under amydriatic conditions. Three ophthalmologists, unaware of the patients’ detailed data, evaluated the photographs using Hyman's classification to evaluate retinal vascular findings. Blood flow in the penile cavernosal artery was measured with colour Doppler ultrasonography, and the peak systolic velocity used as a haemodynamic variable. Correlations among the peak systolic velocity, retinal vascular findings and vascular risk factors (including hypertension, age, cigarette smoking, and hyperlipidaemia) were investigated using multivariate analysis.RESULTSOf the 75 patients, 72 (96%) had both right and left retinal vascular images of sufficient quality for evaluation; 37 were classified as normal and 35 as Grade I, while no patient was Grade II. From a logistic regression multivariate analysis, the peak systolic velocity was the only significant factor correlating with retinal vascular findings, with an odds ratio of 3.34. In contrast, hypertension, age, cigarette smoking and hyperlipidaemia did not correlate significantly with the retinal vascular findings. Similarly, the retinal vascular finding was the only significant factor correlating with the peak systolic velocity of cavernosal blood flow (odds ratio 3.28) and again hypertension, age, cigarette smoking and hyperlipidaemia were not significant factors.CONCLUSIONSThese findings support the assumption that penile erectile function is one of the diseases of atherosclerosis, and emerges nearly simultaneously with retinal vascular disease. It is possible to predict penile arterial conditions in patients with ED from their retinal vascular findings. Thus, amydriatic fundoscopy, a simple practical examination, may be helpful for primary physicians in diagnosing and treating ED.
Ion‐channel currents of smooth muscle cells isolated from the prostate of guinea‐pig
Authors: Oh S.‐J. , Kim K.M. , Chung Y.‐S. , Hong E.‐K. , Shin S.Y. , Kim S.J. 
Published: November 2003

OBJECTIVETo characterize the voltage‐activated ion‐channel currents in guinea‐pig prostate smooth muscle cells (GPSMCs).MATERIALS AND METHODSGPSMCs were isolated using collagenase, and used in a whole‐cell patch clamp study.RESULTSWhen GPSMCs were dialysed with a CsCl solution all the outward K+ currents were blocked and the step‐like depolarization (holding voltage − 70 mV) of the cell membrane evoked inward currents that were completely blocked by nifedipine (1 µmol/L). With KCl solution, step depolarizations showed outward K+ currents composed of fast, transient outward current (Ito) and outward currents that did not inactivate. Ito was resistant to a high concentration of tetraethylammonium (TEA, 5 mmol/L) but was blocked by 4‐aminopyridine (5 mmol/L). The half‐activation and half‐inactivation voltages of Ito were 6 mV and − 58 mV, respectively. With low Ca2+ buffer (0.1 mmol/L EGTA) in the solution, there were spontaneous transient outward currents (STOCs) at depolarized membrane voltages (0 mV). STOCs were blocked by TEA (1 mmol/L) or iberiotoxin (10 nmol/L) but were insensitive to apamin (100 nmol/L).CONCLUSIONThis voltage‐clamp study showed that GPSMCs have l‐type Ca2+ channels and more than two types of K+ channels. The voltage‐ and time‐dependent changes of these ion channels and their interactions might be important in forming action potentials and regulating contractility.
A novel pyrrole derivative, NS‐8, suppresses the rat micturition reflex by inhibiting afferent pelvic nerve activity
Authors: Tanaka M. , Sasaki Y. , Kimura Y. , Fukui T. , Hamada K. , Ukai Y. 
Published: November 2003

OBJECTIVETo investigate the suppressive effect of a newly synthesized compound, 2‐amino‐3‐cyano‐5‐(2‐fluorophenyl)‐4‐methylpyrrole (NS‐8), on micturition, and its mode and sites of action in rats.MATERIALS AND METHODSFemale rats were anaesthetized with urethane, and isovolumetric bladder contractions and cystometrograms recorded. The pelvic afferent discharges from the bladder were also monitored.RESULTSIn the cystometric study, NS‐8 increased the bladder capacity without affecting the maximum bladder contraction pressure, an effect unlike that of currently used anticholinergic drugs for the overactive bladder, which commonly decrease the maximum bladder contraction pressure. Intravesical and intravenous injection of NS‐8 inhibited isovolumetric bladder contractions in a dose‐dependent manner without affecting their amplitude, whereas intracerebroventricular injection with NS‐8 had no such effect. During the urine storage phase of the cystometrogram, NS‐8 decreased the discharge rate of the afferent pelvic nerve from the bladder, in association with a decrease in the increase in intravesical pressure.CONCLUSIONNS‐8 suppressed the micturition reflex by decreasing afferent pelvic nerve activity. Activation of calcium‐sensitive potassium channel of the bladder may be responsible for such an effect. This agent has the potential to treat patients with urinary frequency and incontinence.
Metoidioplasty: a variant of phalloplasty in female transsexuals
Authors: Perovic S.V. , Djordjevic M.L. 
Published: November 2003

OBJECTIVETo describe metoidioplasty, a technique for creating a neophallus from an enlarged clitoris in female transsexuals, without needing the complex, multi‐staged surgical construction of a large phallus, as this reconstruction is one of the most difficult in female transsexuals.PATIENTS AND METHODSFrom September 1995 to April 2002 metoidioplasty was used in 22 patients (aged 18–33 years). The technique is based on the repair of the most severe form of hypospadias and intersex. The ‘urethral plate’ and urethra are completely dissected from the clitoral corporeal bodies, then divided at the level of the glanular corona, and the clitoris straightened and lengthened. A longitudinal vascularized island flap is designed and harvested from the dorsal skin of the clitoris, transposed to the ventral side, tubularized and anastomosed with the native urethra. The new urethral meatus is brought to the top of the neophallus, and the skin of the neophallus and scrotum reconstructed using labia minora and majora flaps.RESULTSThe mean (range) follow‐up was 3.9 (0.5–6) years; the neophallus was 5.7 (4–10) cm, considered satisfactory in 17 patients but the remaining five required additional phalloplasty. The complications were urethral stenosis in two and fistula in three patients.CONCLUSIONSMetoidioplasty is an alternative to phalloplasty, allowing voiding while standing. In patients who desire a larger phallus, various techniques of phalloplasty can also be used.
The prevalence of nocturia and its effect on health‐related quality of life and sleep in a community sample in the USA
Authors: Coyne K.S. , Zhou Z. , Bhattacharyya S.K. , Thompson C.L. , Dhawan R. , Versi E. 
Published: November 2003

OBJECTIVETo evaluate the prevalence of nocturia and its effects, with and without concomitant overactive bladder (OAB, i.e. urgency, frequency, urge incontinence and nocturia), on health‐related quality of life (HRQoL) and sleep.SUBJECTS AND METHODSA national survey was conducted in the USA to assess the prevalence of OAB and nocturia. A nested case‐control study was conducted among respondents with OAB symptoms and age‐ and gender‐matched controls, with participants completing a series of questionnaires on HRQoL (OAB‐q, Short Form‐36, and Medical Outcomes Study (MOS) sleep scale). Descriptive analyses, t‐tests, analysis of variance with post hoc comparisons and multivariate regressions were used to analyse the data.RESULTSIn all, 5204 people participated in the survey, with 919 in the nested case‐control study. The sample population had a mean age of 45.8 years, was 52.6% female and 80% Caucasian. In the community sample, 31% reported > 1 void/night and 14.2% reported > 2 voids/night. The prevalence of nocturia increased with age, with no gender differences. For OAB cases, 66.8% reported > 1 void/night and 42.2% reported > 2. In the case‐control cohort there were significant HRQoL differences (P < 0.01), with increasing episodes of nocturia in all OAB‐q subscales except social interaction. The amount of sleep per night was significantly correlated with the sleep, concern and social interaction OABq subscale scores. The number of nocturia episodes/night was also significantly (P = 0.02) associated with the number of hours of sleep/night.CONCLUSIONNocturia is widely prevalent and increases with age, affecting men and women equally. Incremental increases in the number of voids/night have further negative effects on sleep, symptom bother, and HRQoL.