bluuebunny/crossref_metadata_2025 huggingface dataset:
Orexin 1 Receptor Messenger Ribonucleic Acid Expression and Stimulation of Testosterone Secretion by Orexin-A in Rat Testis
Authors: Barreiro M. L. , Pineda R. , Navarro V. M. , Lopez M. , Suominen J. S. , Pinilla L. , Señaris R. , Toppari J. , Aguilar E. , Diéguez C. , Tena-Sempere M.
Published: February 2004

Orexins are hypothalamic neuropeptides primarily involved in the regulation of food intake and arousal states. In addition, a role for orexins as central neuroendocrine modulators of reproductive function has recently emerged. Prepro-orexin and orexin type-1 receptor mRNAs have been detected in the rat testis. This raises the possibility of additional peripheral actions of orexins in the control of reproductive axis, which remains so far unexplored. To analyze the biological effects and mechanisms of action of orexins in the male gonad, we evaluated testicular expression of orexin receptor 1 (OX1R) and orexin receptor 2 (OX2R) mRNAs in different experimental settings and the effect of orexin-A on testicular testosterone (T) secretion. Persistent expression of OX1R mRNA was demonstrated in the rat testis throughout postnatal development. In contrast, OX2R transcript was not detected at any developmental stage. Expression of OX1R mRNA persisted after selective elimination of mature Leydig cells and was detected in isolated seminiferous tubules at defined stages of the seminiferous epithelial cycle. In addition, testicular OX1R mRNA expression appeared to be under hormonal regulation; it was reduced by long-term hypophysectomy and partially restored by FSH replacement, whereas down-regulation was observed after exposure to increasing doses of the ligand in vitro. Moreover, OX1R mRNA expression was sensitive to neonatal imprinting by estrogen. Finally, orexin-A, in a dosedependent manner, significantly increased basal, but not human choriogonadotropin-stimulated, T secretion in vitro. A similar stimulatory effect was observed in vivo after intratesticular administration of orexin-A. In conclusion, our present results provide the first evidence for the regulated expression of OX1R mRNA and functional role of orexin-A in the rat testis. Overall, our data are suggestive of a novel site of action of orexins in the control of male reproductive axis.
Nitric Oxide Triggers the Phosphatidylinositol 3-Kinase/Akt Survival Pathway in Insulin-Producing RINm5F Cells by Arousing Src to Activate Insulin Receptor Substrate-1
Authors: Tejedo Juan R. , Cahuana Gladys M. , Ramírez Remedios , Esbert Margarida , Jiménez Juan , Sobrino Francisco , Bedoya Francisco J.
Published: February 2004

Mechanisms involved in the protective action of nitric oxide (NO) in insulin-producing cells are a matter of debate. We have previously shown that pharmacological inhibition of c-Src cancels the antiapoptotic action of low and sustained concentrations of exogenous NO. In this study, using insulin-producing RINm5F cells that overexpress Src either permanently active (v-Src) or dominant negative (dn-Src) forms, we determine that this tyrosine kinase is the principal mediator of the protective action of NO. We also show that Src-directed activation of insulin receptor substrate-1, phosphatidylinositol 3-kinase (PI3K), Akt, and Bad phosphorylation conform a substantial component of the survival route because pharmacological inhibition of PI3K and Akt canceled the antiapoptotic effects of NO. Studies performed with the protein kinase G (PKG) inhibitor KT-5823 revealed that NO-dependent activation of c-Src/ insulin receptor substrate-1 is not affected by PKG activation. By contrast, Akt and Bad activation are partially dependent on PKG activation. Endogenous production of NO after overexpression of endothelial nitric oxide synthase in RINm5F cells mimics the effects produced by generation of low amounts of NO from exogenous diethylenetriamine/NO. In addition, we found that NO produces c-Src/PI3K- and PKG-dependent activation of ERK 1/2. The MAPK kinase inhibitor PD 98059 suppresses NO-dependent protection from DNA fragmentation induced by serum deprivation. The protective action of low and sustained concentration of NO is also observed in staurosporine- and Taxol-induced apoptosis. Finally, NO also protects isolated rat islets from DNA fragmentation induced by serum deprivation. These data strengthen the notion that NO production at physiological levels plays a role in protection from apoptosis in pancreatic β-cells.
Specific in vivo phosphorylation sites determine the assembly dynamics of vimentin intermediate filaments
Authors: Eriksson John E. , He Tao , Trejo-Skalli Amy V. , Härmälä-Braskén Ann-Sofi , Hellman Jukka , Chou Ying-Hao , Goldman Robert D.
Published: February 2004

Intermediate filaments (IFs) continuously exchange between a small, depolymerized fraction of IF protein and fully polymerized IFs. To elucidate the possible role of phosphorylation in regulating this equilibrium, we disrupted the exchange of phosphate groups by specific inhibition of dephosphorylation and by specific phosphorylation and site-directed mutagenesis of two of the major in vivo phosphorylation sites determined in this study. Inhibition of type-1 (PP1) and type-2A (PP2A) protein phosphatases in BHK-21 fibroblasts with calyculin-A, induced rapid vimentin phosphorylation in concert with disassembly of the IF polymers into soluble tetrameric vimentin oligomers. This oligomeric composition corresponded to the oligopeptides released by cAMP-dependent kinase (PKA) following in vitro phosphorylation. Characterization of the 32P-labeled vimentin phosphopeptides, demonstrated Ser-4, Ser-6, Ser-7, Ser-8, Ser-9, Ser-38, Ser-41, Ser-71, Ser-72, Ser-418, Ser-429, Thr-456, and Ser-457 as significant in vivo phosphorylation sites. A number of the interphase-specific high turnover sites were shown to be in vitro phosphorylation sites for PKA and protein kinase C (PKC). The effect of presence or absence of phosphate groups on individual subunits was followed in vivo by microinjecting PKA-phosphorylated (primarily S38 and S72) and mutant vimentin (S38:A, S72:A), respectively. The PKA-phosphorylated vimentin showed a clearly decelerated filament formation in vivo, whereas obstruction of phosphorylation at these sites by site-directed mutagenesis had no significant effect on the incorporation rates of subunits into assembled polymers. Taken together, our results suggest that elevated phosphorylation regulates IF assembly in vivo by changing the equilibrium constant of subunit exchange towards a higher off-rate.
The surface of articular cartilage contains a progenitor cell population
Authors: Dowthwaite Gary P. , Bishop Joanna C. , Redman Samantha N. , Khan Ilyas M. , Rooney Paul , Evans Darrell J. R. , Haughton Laura , Bayram Zubeyde , Boyer Sam , Thomson Brian , Wolfe Michael S. , Archer Charles W.
Published: February 2004

It is becoming increasingly apparent that articular cartilage growth is achieved by apposition from the articular surface. For such a mechanism to occur, a population of stem/progenitor cells must reside within the articular cartilage to provide transit amplifying progeny for growth. Here, we report on the isolation of an articular cartilage progenitor cell from the surface zone of articular cartilage using differential adhesion to fibronectin. This population of cells exhibits high affinity for fibronectin, possesses a high colony-forming efficiency and expresses the cell fate selector gene Notch 1. Inhibition of Notch signalling abolishes colony forming ability whilst activated Notch rescues this inhibition. The progenitor population also exhibits phenotypic plasticity in its differentiation pathway in an embryonic chick tracking system, such that chondroprogenitors can engraft into a variety of connective tissue types including bone, tendon and perimysium. The identification of a chondrocyte subpopulation with progenitor-like characteristics will allow for advances in our understanding of both cartilage growth and maintenance as well as provide novel solutions to articular cartilage repair.
Endocytic trafficking in actively resorbing osteoclasts
Authors: Stenbeck Gudrun , Horton Michael A.
Published: February 2004

Endocytosis and the subsequent intracellular trafficking of the endocytosed material are important determinants of cellular function. Osteoclasts, cells of the monocyte/macrophage family, are specialized for the internalization and processing of bone matrix. Transcytosis of endocytosed material has been observed in osteoclasts but the precise mechanism controlling this process is unclear. Here, we investigate the regulation of these trafficking events. To establish the directionality and kinetics of trafficking events in resorbing osteoclasts, we devised a system using fluorescent low-molecular-weight markers as probes to follow the route taken by the digested bone matrix. We demonstrate that this route is largely distinct from the pathway followed by proteins taken up by receptor-mediated endocytosis at the basolateral plasma membrane. Endocytosis and transcytosis from the ruffled border are fast processes, with a half-life of the endocytosed material inside the cells of 22 minutes. We demonstrate the crucial role of the microtubule network in transport from the ruffled-border area and provide evidence for a role of the cytoskeleton in the overall efficacy of trafficking. Moreover, we analyse the effect of the V-ATPase inhibitor bafilomycin A1 on endocytic uptake, which gives insight into the pH-dependent regulation of membrane trafficking and resorption in osteoclasts.
The amount of neurofilaments aggregated in the cell body is controlled by their increased sensitivity to trypsin-like proteases
Authors: Fasani F. , Bocquet A. , Robert P. , Peterson A. , Eyer J.
Published: February 2004

Neurofilaments are synthesised and assembled in neuronal cell bodies, transported along axons and degraded at the synapse. However, in several pathological situations they aggregate in cell bodies or axons. To investigate their turnover when separated from their normal site of degradation, we used a previously described transgenic model characterised by perikaryal retention of neurofilaments, and compared the basic features of both neurofilament synthesis and degradation with that observed in normal mice. Despite the massive perikaryal aggregates, neurofilament transcript levels were found to be unchanged, whereas the total accumulation of neurofilament proteins was markedly reduced. Neurofilaments isolated from transgenic samples are more sensitive to both trypsin and α-chymotrypsin mediated proteolysis. Consistent with their greater in vitro sensitivity, trypsin immunolabeling of cell bodies was stronger in transgenic mice. These results show a novel mechanism to regulate the amount of neurofilaments when they abnormally aggregate.
Evidence for structural and functional diversity among SDS-resistant SNARE complexes in neuroendocrine cells
Authors: Kubista Helmut , Edelbauer Hannah , Boehm Stefan
Published: February 2004

The core complex, formed by the SNARE proteins synaptobrevin 2, syntaxin 1 and SNAP-25, is an important component of the synaptic fusion machinery and shows remarkable in vitro stability, as exemplified by its SDS-resistance. In western blots, antibodies against one of these SNARE proteins reveal the existence of not only an SDS-resistant ternary complex but also as many as five bands between 60 and >200 kDa. Structural conformation as well as possible functions of these various complexes remained elusive.

In western blots of protein extracts from PC12 cell membranes, an antibody against SNAP-25 detected two heat-sensitive SDS-resistant bands with apparent molecular weights of 100 and 230 kDa. A syntaxin antibody recognized only the 230 kDa band and required heat-treatment of the blotting membrane to detect the 100 kDa band. Various antibodies against synaptobrevin failed to detect SNARE complexes in conventional western blots and detected either the 100 kDa band or the 230 kDa band on heat-treated blotting membranes.

When PC12 cells were exposed to various extracellular K+-concentrations (to evoke depolarization-induced Ca2+ influx) or permeabilized in the presence of basal or elevated free Ca2+, levels of these SNARE complexes were altered differentially: moderate Ca2+ rises (≤1 μM) caused an increase, whereas Ca2+ elevations of more than 1 μM led to a decrease in the 230 kDa band. Under both conditions the 100 kDa band was either increased or remained unchanged.

Our data show that various SDS-resistant complexes occur in living cells and indicate that they represent SNARE complexes with different structures and diverging functions. The distinct behavior of these complexes under release-promoting conditions indicates that these SNARE structures have different roles in exocytosis.
Gap junctional communication modulates agonist-induced calcium oscillations in transfected HeLa cells
Authors: Lin George C. , Rurangirwa Jacqueline K. , Koval Michael , Steinberg Thomas H.
Published: February 2004

Gap junctional communication modulates intercellular calcium signaling in many cell types. We have investigated whether gap junctional communication modulates calcium oscillatory behavior of cells responding to an agonist. Extracellular UTP induced calcium oscillations in 70% of HeLa cells cultured in monolayer, and neighboring cells oscillated independently of each other. In HeLa cell transfectants expressing connexin43 (HeLa/Cx43), extracellular UTP induced calcium transients, but calcium oscillations occurred in only 10% of cells. Inhibition of gap junctional communication with anandamide in HeLa/Cx43 transfectants substantially restored oscillations (55% of cells). In HeLa/Cx45 transfectants, UTP initiated calcium oscillations similar to those seen in HeLa cells (63% of cells), but HeLa/Cx46 transfectants demonstrated calcium oscillations that were dampened compared to those of the parental HeLa cells, and occurred in only 40% of cells. These experiments demonstrate that gap junctional communication modulates calcium oscillatory behavior in cell monolayers, presumably by allowing cells to share a small molecule such as inositol trisphosphate. These studies suggest that gap junctional communication may alter the nature of signals induced by calcium mobilizing agonists in a connexin-dependent fashion by modulating calcium oscillatory behavior.
A novel RING-finger-like protein Ini1 is essential for cell cycle progression in fission yeast
Authors: Oltra Elisa , Verde Fulvia , Werner Rudolf , D'Urso Gennaro
Published: February 2004

We have cloned a fission yeast (Schizosaccharomyces pombe) homologue of Ini, a novel RING-finger-like protein recently identified in rat that interacts with the connexin43 (cx43) promoter and might be important for the response of the cx43 gene to estrogen. S. pombe cells deleted for ini1+ fail to form colonies and arrest with an elongated cell phenotype, indicating a cell cycle block. Cell cycle arrest is dependent on expression of Wee1, but not Rad3, suggesting that it occurs independently of the DNA damage checkpoint control. Analysis of mRNA intermediates in cells depleted for Ini1 demonstrates that Ini1 is required for pre-mRNA splicing. We observe an accumulation of pre-mRNA for six of seven genes analysed, suggesting that Ini1 is required for general splicing activity. Interestingly, loss of Ini1 results in cell death that is partially suppressed by elimination of the Wee1 kinase. Therefore, Wee1 might promote cell death in the absence of Ini1.
Cyclobutadiene Dianion
Authors: Matsuo Tsukasa , Sekiguchi Akira
Published: February 2004

In this account, we present the synthesis and chemistry of the dilithium salts of the cyclobutadiene dianion (CBD2−), which contains a doubly charged four-membered ring system. The transmetalation reaction of tetrasilyl-substituted cyclobutadiene (CBD) cobalt complexes (5,6) with lithium metal led to the formation of the corresponding dilithium salts of CBD2− (10,11). The aromaticity of the CBD2− species has been studied on the basis of structural characteristics and magnetic properties. The dilithium salts of CBD2− stabilized by both phenyl and silyl groups (12–14) were also prepared by a similar procedure, starting from the corresponding CBD cobalt complexes (7–9). The CBD2− species have been fully characterized by NMR studies and X-ray crystallography. The two-electron oxidation reaction of the tetrasilyl-CBD2− (10,11) with 1,2-dibromoethane produced the neutral CBD derivatives (17,22). The structures, properties, and reactivities of the tetrasilyl-substituted CBD are also reviewed here. Tetrakis(trimethylsilyl)tetrahedrane (26) was synthesized by irradiation of tetrakis(trimethylsilyl)cyclobutadiene (17). The structure and properties of tetrahedrane (26) are presented.
Optically Active Polymers for Chiral Separation
Authors: Yamamoto Chiyo , Okamoto Yoshio
Published: February 2004

One of the most practically important functions of optically active polymers is chiral recognition. In the past two decades, various optically active polymers have been utilized as the chiral stationary phases (CSPs) for high-performance liquid chromatography, which is today the most powerful and practical method not only for analyzing enantiomers but also for obtaining them in a pure form. In this account, the emphasis lies on the polymethacrylate and polysaccharide derivatives that we have developed. One-handed helical poly(triphenylmethyl methacrylate) has been prepared by asymmetric anionic polymerization and shows high chiral recognition, especially for the compounds which have no functional groups. Polysaccharide derivatives, particularly cellulose and amylose phenylcarbamates, can resolve a wide range of racemates, and their abilities are significantly influenced by the substituents introduced on the phenyl moieties. Cellulose and amylose cycloalkylcarbamates and chitin phenylcarbamates also show high chiral recognition; the former can be utilized as the CSPs for thin-layer chromatography because of the absence of UV absorption. The immobilization of the polysaccharide derivatives onto a silica support via chemical bonding improves their durability for various solvents and also their resolving ability for some compounds. Several phenylcarbamates are soluble in chloroform, which allowed one to study their chiral recognition mechanism at a molecular level, particularly by NMR spectroscopy. A computational method can also be applied for mechanistic studies.
ESR Spin Probe Investigation of Chain Ordering of a Triglycerol Membrane
Authors: Nakagawa Kouichi
Published: February 2004

The chain ordering of a triglycerol membrane was investigated by continuous wave (CW) electron spin resonance (ESR) in conjunction with a spin probe method. Order parameters (S0) obtained by a slow-tumbling computational analysis for 7-doxylstearic acids (7-DSA), 12-DSA, and 16-DSA in poly(oxyethylene) hydrogenated castor oil (HCO) membrane at 20 °C were 0.68, 0.25, and 0.04, respectively. The S0 results suggest that the membrane chains are well ordered around the probe moiety for 7-DSA and very fluid around the 16-DSA moiety in the HCO membrane. The relative magnitude of the chain order for 12-DSA was very different from that obtained by the conventional order parameter using hyperfine coupling values. The conventional hyperfine analysis cannot verify the subtle spectral changes for 12-DSA. Moreover, the S0 values for 7- and 12-DSA decreased differently when the temperature increased. Thus, the results obtained by a slow-tumbling simulation revealed detailed membrane chain ordering.
Three Types of Nickel(II) Complexes Derived from 2-Substituted Benzothiazoline; Formation of a Tetranuclear Complex by a Sterically Induced Orthometallation Reaction
Authors: Kawamoto Tatsuya , Kushi Yoshihiko
Published: February 2004

The reaction of 2-(2,4-dichlorophenyl)benzothiazoline with nickel(II) acetate tetrahydrate in ethanol has afforded the monohelical bis[2-(2,4-dichlorophenylmethyleneamino)benzenethiolato]nickel(II) ([Ni(HL1)2] (1)). The X-ray structure of 1 has revealed that ortho-phenyl hydrogen atoms of pendant arms are oriented to the nickel center in a monohelical structure. The low-field resonance for their ortho-protons in the 1H NMR spectrum indicates that the Ni···H–C interaction is maintained in solution. Heating 1 in toluene gave a square-planar nickel(II) complex with a non-innocent ligand ([Ni(L2)] (2)), and prolonged heating yielded a thiolato-bridged tetranuclear nickel(II) complex ([Ni4(L1)4] (3)). The tetranuclear complex 3 was formed from activation of the ortho C–H bond, not the C–Cl bond, through sterically controlled Ni···H–C contacts in 1. The molecular structures of 2 and 3 have also been determined by X-ray crystallographic methods.
Determination of the Toxic Lead Level in Cosmetic-Hair Dye Formulations Using a Screen-Printed Silver Electrode
Authors: Shih Ying , Zen Jyh-Myng , Senthil Kumar Annamalai , Lee Yi-Chien , Huang Hui-Rou
Published: February 2004

A disposable-type screen-printed non-single crystal silver electrode (AgSPE) surface has been demonstrated for the sensitive and selective quantification of lead in cosmetic hair-dye formulations using its underpotential deposition (UPD-Pb) process.
Exciplex Emission of Carbazole–Terephthalate Cyclophane and Its Application as a Fluorescent Probe for Local Polarity of Solvents
Authors: Benten Hiroaki , Ohkita Hideo , Ito Shinzaburo , Yamamoto Masahide , Tohda Yasuo , Tani Keita
Published: February 2004

Photophysical properties of carbazole–terephthalate cyclophane (Cz–TP) were studied. The absorption spectra of Cz–TP in various solvents were slightly broadened due to the distortion of the carbazole (Cz) moiety and became similar to that of a reference compound of carbazole–benzene cyclophane (Cz–Bz). The absorption spectrum and the peak wavelengths of Cz–TP were insensitive to the solvent polarity. These findings show that there is transannular π–π electronic interaction between the Cz and TP moieties but that the charge transfer interaction from the Cz to TP moieties is small in the ground state. On the other hand, the emission spectrum was broad, structureless, and red-shifted compared with that of the reference compound. The peak wavelength of the emission was shifted to a longer wavelength with increases in the dielectric constant of solvents. The peak shift was quantitatively explained by the Lippert–Mataga formula. Therefore, we conclude that the broad emission is due to the intramolecular exciplex in Cz–TP. The dielectric constant of 2-methyl tetrahydrofuran was evaluated from the exciplex emission of Cz–TP ranging from 77 to 292 K, indicating that the Cz–TP molecule is an excellent polarity probe not only in a liquid phase but also in a solid phase over a wide temperature range.
Explaining gay men's unrealistic optimism about becoming infected with HIV
Authors: Gold R S
Published: February 2004

Among the self-justifications that gay men use when deciding to have unprotected intercourse is the thought that they are at less risk than most gay men. Two explanatory models of such 'unrealistic optimism' (UO) have been proposed: while the motivational account holds that UO serves the function of bringing comfort, the cognitive account holds that UO serves no particular function, being simply a by-product of normal cognitive strategies. This study tested the prediction, derived from the motivational account, that highlighting the salience of the self-other comparison should increase UO. Gay men uninfected with HIV ( n = 122) estimated both their own risk of contracting various health problems - among them, becoming infected with HIV - and that of the average gay man. The purported aim of collecting the data was varied, so as to either make the self-other comparison central to the aim or render one of the two types of estimate irrelevant to the aim. No effect on UO was found. It seems that the cognitive account provides a better explanation than does the motivational account of at least that form of UO measured in this study. Implications for AIDS education are discussed.
Prevalence of sexually transmitted diseases and human immunodeficiency virus among women attending prenatal services in Apia, Samoa
Authors: Sullivan E A , Koro Semo , Tabrizi S , Kaldor J , Poumerol G , Chen S , O'Leary M , Garland S M
Published: February 2004

There is no routine prenatal screening for sexually transmitted diseases (STDs) and human immunodeficiency virus (HIV) in pregnancy in Samoa. Testing for chlamydial infection is not available. To gather information on pregnant women, a prevalence survey was conducted in Apia, Samoa, utilizing two prenatal hospital clinics. Pregnant ( n = 427) women were tested for Neisseria gonorrhoeae, Chlamydia trachomatis and Trichomonas vaginalis using polymerase chain reaction (PCR), and for syphilis ( n = 441) by rapid plasmid reagin (RPR) and HIV ( n = 441) by enzyme-linked immunosorbent assay (ELISA). Results were: chlamydia 30.9% (132); trichomoniasis 20.8%; gonorrhoea 3.3%; syphilis 0.5%; and HIV 0%. Overall 42.7% had at least 1 STD. Young women aged <25 years were three times more likely to have a STD than older women (odds ratio = 3.0, 95% confidence intervals 2.0, 4.5). The lack of inexpensive, reliable field diagnostics remain a barrier to sustainable STD control programmes for pregnant women living in developing countries.
Post-exposure prophylaxis for human immunodeficiency virus infection in the Royal Infirmary of Edinburgh - an audit
Authors: Manavi K , McMillan A , Paterson J
Published: February 2004

The purpose of the audit was to assess the implementation of the protocols developed by Lothian University National Health Service Trust for post-exposure prophylaxis (PEP) against blood-borne virus infections.

Patients at risk of infection after an injury were referred to the Department of Genitourinary Medicine (GUM) Edinburgh Royal Infirmary for assessment. A brief description of the incident, the time of the incident, the time of referral and the decision made was documented. The notes of these patients were reviewed 12 months later for the results of prospective HIV test after each incident.

There were 76 referrals to the GUM Department in year 2001. Occupational Health (OH) and the Accident and Emergency (A&E) Departments each made 35 referrals. The latter cases were exposures in community settings. Amongst A&E referrals the time interval between the incident and assessment was significantly longer than for OH referrals ( P = 0.001). Female health workers reported exposure incidents sooner than their male counterparts ( P = 0.01). Post-exposure prophylaxis for HIV was offered to 22 (29%) of 76 referred individuals, and was accepted by 13 (59%) persons. The course of PEP drugs was discontinued by five (38%) of the 13 individuals after consented postincident HIV testing of the source patients showed that they were HIV-seronegative. Five of eight individuals completed the course of PEP medication. Only 11 (14%) persons attended for serological testing for blood-borne virus infection three or more months after exposure. None of these 11 persons, one of whom had been given PEP medication, had become infected.

Further training and emphasis on the necessity of immediate reporting of occupational incidents and consented post-incident HIV testing of the source patient is needed. The current guidelines appear to work fairly well, but further counselling of those with significant injuries to undertake HIV serological testing after appropriate time interval is required.
Remission of Behçet's disease and keratoconjunctivitis sicca in an HIV-infected patient treated with HAART
Authors: Cicalini S , Gigli B , Palmieri F , Boumis E , Froio N , Petrosillo N
Published: February 2004

A 34-year-old woman presented with a 10-year history of recurrent oral and genital ulcerations and recurrent episodes of bilateral conjunctivitis associated with HIV infection. A diagnosis of Behçet's disease (BD) in association with keratoconjunctivitis sicca (KCS) was made after exclusion of other viral and autoimmune diseases according to the international criteria for BD.

This is the first reported case of a combination of BD and KCS in a patient with HIV infection in which a complete resolution was observed as a result of successful highly active antiretroviral therapy. The likelihood that a direct viral effect or HIV-induced autoimmune mechanisms act in the pathogenesis of both BD and KCS in HIV-infected patients is discussed.
Extramammary Paget's disease masquerading as lichen sclerosus
Authors: Bansal D , Bowman C A
Published: February 2004

A wide range of genital infections and skin conditions may present with vulval pruritus. Lichen sclerosus is one cause. This is often associated with visible skin changes which include atrophy, resorption or fusion of the labia, sclerosis and excoriation. Ulceration may indicate malignant transformation and requires urgent biopsy.

Many experienced clinicians diagnose lichen sclerosus on clinical appearance, however we recommend a low threshold for biopsy, especially for non-responding or odd looking lesions. We report a patient whose vulval skin biopsy for clinically suspected lichen sclerosus revealed extramammary Paget's disease of the vulva. We review the pathology, prognosis and treatment of this condition.
A review of telemedicine in Romania
Authors: Panait Lucian , Doarn Charles R , Saftoiu Adrian , Popovici Calin , Valeanu Vlad , Merrell Ronald C
Published: February 2004

Romania is an eastern European country that is undergoing rapid reform of its medical system. We conducted an assessment of the potential for telemedicine in the country, through a literature review, personal visits to Romania and discussions with individuals from academia, the Ministry of Health and Family, and businesses. The results suggest that telemedicine has the potential to accelerate health-care reform. The main hospitals and universities could promote the wider distribution and development of telemedicine within Romania, which in turn would bring benefits to the Romanian people, 46% of whom live in rural areas.
The challenges of following patients and assessing outcomes in teledermatology
Authors: Krupinski Elizabeth A , Engstrom Mike , Barker Gail , Levine Norman , Weinstein Ronald S
Published: February 2004

We retrospectively reviewed the follow-up and outcomes of 50 store-and-forward teledermatology patients, and compared the findings with those from a control group of 50 patients who had been seen in person. Patient records were examined for a six-month period following the initial referral to a dermatologist. Variables examined included medical records from the referral, evidence of actions taken (e.g. biopsy), evidence of follow-up visits, and what (if any) clinical outcomes were noted. There were few differences between the teledermatology and in-person groups. The main difference was whether there was any report in the record that the referring clinician took some action based on the consultation with the specialist: there were more reports of action being taken in the teledermatology group than in the in-person group. Reports of outcomes were found in only 6% and 8% of the records of the teledermatology and in-person groups, respectively. The challenges of assessing outcomes in teledermatology for rural patients include patient loss to follow-up, lack of information in the patient records and low rates of patient return to the referring clinician for follow-up.
Patient satisfaction with teledermatology: quantitative and qualitative results from a randomized controlled trial
Authors: Collins Karen , Walters Stephen , Bowns Ian
Published: February 2004

As part of a randomized controlled trial involving 208 dermatology patients, a quantitative and qualitative study was undertaken to explore patients' satisfaction with a specialist dermatological opinion and further management obtained through either a traditional outpatient consultation (control group) or an asynchronous teleconsultation (telemedicine group). There was a response rate of 71% to the quantitative patient satisfaction survey (148 replies from 208 distributed questionnaires). The responders comprised 80 of the 111 telemedicine patients (72%) and 68 of the 97 control patients (70%). Overall levels of patient satisfaction were high in both groups, and there was no significant difference between them. Ninety per cent of patients in the control group were satisfied with their overall care, compared with 81% in the telemedicine group, and 87% of patients in the control group were satisfied with their overall management, compared with 84% in the telemedicine group. Follow-up qualitative interviews with 30 of the participants also suggested that patients were generally positive about their care and management, regardless of group, age or gender. Receiving a diagnosis, treatment and cure, receiving adequate information and explanations, the need to be taken seriously, the need for individualized personal care, and the importance of a short waiting time for an appointment and treatment were all aspects of care and management most likely to result in patient satisfaction, regardless of modality.
A pilot study of a combined dermoscopic–pathological approach to the telediagnosis of melanocytic skin neoplasms
Authors: Ferrara Gerardo , Argenziano Giuseppe , Cerroni Lorenzo , Cusano Francesco , Di Blasi Arturo , Urso Carmelo , Soyer H Peter
Published: February 2004

We examined a combined (dermoscopic–pathological) approach to the telediagnosis of melanocytic skin lesions. A store-and-forward teleconsultation was simulated. Dermoscopic and histopathology images from 12 melanocytic lesions were stored in a telepathology workstation. A dermoscopy consultant, a histopathology consultant and an expert in dermoscopic–pathological correlation gave their diagnoses and comments on the images. The consensus diagnosis between two teleconsultants on the original histological slides was regarded as the gold standard. The diagnostic accuracy was 83% (including one false negative diagnosis of malignancy) for teledermoscopy and 100% for teledermatopathology. The combined approach detected one case that showed a much greater atypia on dermoscopy than on histopathology. In this case step-sections of the sample were deemed to be required for definite diagnosis. The combined approach was helpful in detecting macroscopic and microscope sampling errors of melanocytic lesions during teleconsultation.
A pilot study of health education via a nurse-run telephone self-management programme for elderly people with osteoarthritis
Authors: Blixen Carol E , Bramstedt Katrina A , Hammel Jeffrey P , Tilley Barbara C
Published: February 2004

We conducted a pilot study of a nurse-run telephone self-management programme for elderly people with osteoarthritis (OA). Thirty-two subjects, aged 60 years or more, with a diagnosis of OA were recruited from two hospital rheumatology clinics and were randomized to a control or intervention group. The intervention group received six weekly mailings of OA health education modules, a relaxation audio-tape and six weekly 45 min follow-up telephone self-management sessions. There was a 100% compliance rate in the intervention group, and all subjects completed baseline and three-month interviews; one subject in each group was lost to six-month follow-up. There were no significant differences in self-management between the control and intervention groups. However, at three months there were improvements in the intervention group (relative to baseline) on some outcome measures. The results suggest that the telephone may be a useful tool for reinforcing health-promoting activities for patients.
Telephone advice nursing services in a US health maintenance organization
Authors: Shapiro Susan E , Izumi Shigeko , Tanner Christine A , Moscato Susan R , Valanis Barbara G , David Marion R , Gullion Christina M
Published: February 2004

We studied telephone advice nursing (TAN) in the US. We recorded 4269 TAN calls in four regions served by Kaiser Permanente, a health maintenance organization. A call description form was used to record information regarding both calls and callers. The mean call length was 5.9 min (SD 3.6, range 0.3-35.8); 300 calls lasted less than 2 min. The mean call length differed significantly across regions, from 4.4 min in Hawaii to 8.7 min in Southern California. Calls to call centres lasted an average of 6.5 min (SD 3.8), compared with 4.2 min (SD 2.9) for those to medical offices. These differences were significant. Although 42% of calls required some further medical management, only 18% ( n = 754) resulted in an urgent disposition, and only 16% ( n = 121; 3% of all calls) of the urgent dispositions involved referral to emergency services. The likelihood of urgent disposition varied significantly by region. Callers generally used the TAN services for the right reasons, that is, with questions or concerns that could be reasonably handled by telephone advice nurses.
Early Experiences and Clinical Implications of Drug-Eluting Stents: Part 1
Authors: Chong Pang H , Cheng Judy WM
Published: February 2004

OBJECTIVE

To review the pathogenesis of in-stent restenosis and the evolution of drug-eluting stents (DES).

DATA SOURCES

Using the search terms sirolimus, paclitaxel, and drug-eluting stents, a literature review was conducted to identify peer-reviewed articles and abstracts in MEDLINE (1966—June 2003). Recent meeting abstracts were accessed through the American Heart Association and the American College of Cardiology Web sites. Citations from available articles were also reviewed for additional references.

STUDY SELECTION AND DATA EXTRACTION

Published reviews and studies showing the effects of in-stent restenosis and drug-coated and -eluting stents were evaluated and reviewed.

DATA SYNTHESIS

Coronary stent implantation is a common form of percutaneous coronary interventions. Approximately 20–30% of patients undergoing stent placement develop restenosis 6 months after the procedure. The primary cause of in-stent restenosis is neointimal proliferation and subsequent accumulation of extracellular matrix, collagen, and macrophages. Eluting stents with an antimitotic agent may reduce the extent of restenosis. Research is ongoing in terms of finding the ideal combination of antimitotic agent, stent, and eluting medium to create the perfect stent.

CONCLUSIONS

Research over the last decade has led to a better understanding of the pathogenesis of in-stent restenosis and ways to prevent restenosis. DES are being developed as one of the strategies to prevent restenosis.
Inert Medication Ingredients Causing Nonadherence Due to Religious Beliefs
Authors: Sattar S Pirzada , Ahmed Mohammed Shakeel , Majeed Farhan , Petty Frederick
Published: February 2004

OBJECTIVE

To report 4 cases of medication nonadherence due to presence of inert ingredients forbidden by the patients’ religion.

CASE SUMMARIES

We describe 4 cases in which religious concerns about prescribed medications’ inert components led to discontinuation of these medications. These inert components are gelatin and stearic acid, which might be derived from pork or beef products. In these 4 cases, patients of Muslim, Orthodox Christian, and Seventh Day Adventist faiths, who consider it against their religion to consume pork products, stopped their medications on discovering this possibility. This led to relapse of their illnesses.

DISCUSSION

These cases demonstrate that, for some patients, inert medication components that are forbidden by their religion may lead to discontinuation of medications. This could lead to relapse of symptoms and might even lead to hospitalization. Therefore, it is important for prescribers to inform patients of this possibility when treating patients whose religious background might conflict with these inert medication components.

CONCLUSIONS

Patients with religion prohibitions against consumption of pork and/or beef products might stop their medications when prescribed those with pork- and beef-derived gelatin and/or stearic acid. Prescribers should discuss this possibility with their patients, perhaps as part of informed consent.
Improvement in Sensitivity of Allele-specific PCR Facilitates Reliable Noninvasive Prenatal Detection of Cystic Fibrosis
Authors: Nasis Ourania , Thompson Shanel , Hong Tom , Sherwood Margaret , Radcliffe Shawn , Jackson Laird , Otevrel Tomas
Published: February 2004

Background: Cell-free fetal DNA circulating in maternal blood has potential as a safer alternative to invasive methods of prenatal testing for paternally inherited genetic alterations, such as cystic fibrosis (CF) mutations.

Methods: We used allele-specific PCR to detect mutated CF D1152H DNA in the presence of an excess of the corresponding wild-type sequence. Pfx buffer (Invitrogen) containing replication accessory proteins and Taq polymerase with no proofreading activity was combined with TaqMaster PCR Enhancer (Eppendorf) to suppress nonspecific amplification of the wild-type allele. The procedure was tested on DNA isolated from plasma drawn from 11 pregnant women (gestational age, 11–19.2 weeks), with mutation confirmation by chorionic villus sampling.

Results: The method detected 5 copies of the CF D1152H mutant allele in the presence of up to ∼100 000 copies of wild-type allele without interference from the wild-type sequence. The D1152H mutation was correctly identified in one positive sample; the only false-positive result was seen in a mishandled sample.

Conclusions: This procedure allows for reliable detection of the paternally inherited D1152H mutation and has potential application for detection of other mutations, which may help reduce the need for invasive testing.
Analytic Performance of Immunoassays for Drugs of Abuse Below Established Cutoff Values
Authors: Luzzi Veronica I , Saunders Al N , Koenig John W , Turk John , Lo Stanley F , Garg Uttam C , Dietzen Dennis J
Published: February 2004

Background: The analytic performance and accuracy of drug detection below Substance Abuse and Mental Health Services Administration (SAMHSA) cutoffs is not well known. In some patient populations, clinically significant concentrations of abused drugs in urine may not be detected when current SAMHSA cutoffs are used. Our objectives were to define the precision profiles of three immunoassay systems for drugs of abuse and to evaluate the accuracy of testing at concentrations at which the CV was <20%.

Methods: Drug-free urine was supplemented with analytes to assess the precision in three commercial drugs-of-abuse immunoassay systems below the SAMHSA-dictated cutoffs for amphetamines, opiates, benzoylecgonine, phencyclidine, and cannabinoids. Consecutive urine samples with signals associated with a CV <20% by Emit® immunoassay and below SAMHSA cutoffs were then subjected to confirmatory analysis.

Results: The CV of all immunoassay systems tested remained <20% to drug concentrations well below SAMHSA cutoffs. The accuracy of urine drug-screening results between the SAMHSA-specified cutoffs and the precision-based cutoffs was less than accuracy for specimens above the SAMHSA cutoffs, but the use of the precision-based cutoff produced a 15.6% increase in the number of screen-positive specimens and a 7.8% increase in the detection of specimens that yielded positive results on confirmatory testing.

Conclusion: The precision of three commercial immunoassay systems for drugs-of-abuse screening is adequate to detect drugs below SAMHSA cutoffs. Knowledge of the positive predictive values of screening immunoassays at lower cutoff concentrations could enable efficient use of confirmatory testing resources and improved detection of illicit drug use.
Development of an Immunofluorometric Assay and Quantification of Human Kallikrein 7 in Tissue Extracts and Biological Fluids
Authors: Kishi Tadaaki , Soosaipillai Antoninus , Grass Linda , Little Sheila P , Johnstone Edward M , Diamandis Eleftherios P
Published: February 2004

Background: Human kallikrein 7 (hK7), also known as human stratum corneum chymotryptic enzyme, is a chymotrypsin-like serine protease first identified in human skin extracts and predicted to be a secreted protease. The aim of this study was to develop a sensitive and specific immunoassay for hK7 and to examine the distribution of hK7 in tissue extracts and biological fluids.

Methods: Recombinant hK7 was produced in human embryonic kidney cells (HEK293T) and purified by a three-step column chromatographic procedure. The purified hK7 was injected into mice for antibody generation. A sandwich-type immunoassay was developed with the anti-hK7 monoclonal antibodies.

Results: The assay had imprecision (CV) <10% through the dynamic range of 0.2–20 μg/L and had no detectable cross-reactivity from other members in the human kallikrein gene family. Highest concentrations were found in skin, esophagus, and kidney. hK7 was also found in amniotic fluid, ascites from ovarian cancer patients, breast milk, cerebrospinal fluid, saliva, seminal plasma, serum, sweat, synovial fluid, and urine.

Conclusions: This study describes the first ELISA-type immunoassay for hK7 protein quantification. hK7 is found many human tissues and in various biological fluids.
Learning about Foreign Markets: Are Entrant Firms Exposed to a “Shock Effect”?
Authors: Pedersen Torben , Petersen Bent
Published: February 2004

This study addresses how managers’ perceived familiarity with local markets develops during a period of entry or expansion. The authors derive different predictions of how foreign-market familiarity changes during periods of entry from the internationalization process literature. They then subject the predictions to an empirical examination, using a data set that covers foreign business operations as reported by managers of international firms from small, open economies (Denmark, Sweden, and New Zealand). In addition to addressing these predictions, the empirical study gives insight into the incidence and character of “shock effects” in foreign-market entries. The effects are evident in managers’ inclination to underestimate differences between the home and host countries in terms of business environments. The data support the supposition that, in general, managers of entrant firms experience these shock effects. For example, the authors find the lowest level of market familiarity among managers of entrant firms that are in the eighth year after entry or initiation of foreign-market expansion. The company data indicate that, in general, managers of entrant firms experience shock effects in relation to entry into adjacent but not into distant countries. Thus, the hypothesis of the psychic-distance paradox that the authors put forward is supported. Entrant firms also experience shock effects with respect to the acquisition of tacit rather than explicit knowledge; furthermore, the data suggest that shock effects befall producers of customized products but not producers of standardized products.
Mathematical comparison between volume of distribution (V) and volume of distribution at steady‐state (Vss) utilizing model‐independent approach
Authors: Sobol Eyal , Bialer Meir
Published: February 2004

Pharmacokinetic textbooks state that the (apparent) volume of distribution based on drug concentration in plasma (V or Vβ) is always greater than the volume of distribution (apparent) under steady state conditions (Vss), but do not provide a general model‐independent mathematical proof. Wagner's mathematical comparison between Vβ and Vss is based on microscopic rate constants of either specific models and is restricted solely to the two‐compartment open body model. Nakashima and Benet utilizing a model‐dependent approach showed a mathematical relationship between Vβ and Vss for a multicompartment model, but again by using microscopic model constants. The limitation of these two above mentioned mathematical comparisons is the necessity of knowledge of the model's structure and its microscopic rate constants. The present article describes a new non‐compartmental, model‐independent, general mathematical proof for Vβ to be always greater than Vss. This new method does not require any knowledge of microscopical rate constants and is based solely on an exponentially decreasing function, which is the common way to describe drug disposition following i.v. bolus. Copyright © 2004 John Wiley & Sons, Ltd.