bluuebunny/crossref_metadata_2025 huggingface dataset:
Expression and Characterization of the Reverse Transcriptase Enzyme from Type 1 human Immunodeficiency Virus Using Different Baculoviral Vector Systems
Authors: Pekrun Katja , Petry Harald , Jentsch Klaus‐Dieter , Moosmayer Dieter , Hunsmann Gerhard , Lüke Wolfgang
Published: December 2004

To produce the human immunodeficiency virus type 1 (HIV‐1) reverse transcriptase (RT) in amounts required to study its structure and function, the p66 enzyme subunit was expressed using two different baculovirus vectors in Sf158 insect host cells. Both vectors permitted an efficient HIV‐1 RT expression. The resulting products were purified up to 90% homogeneity, characterized, and investigated for their susceptibility to digestion with various proteases.

The recombinant baculoviral RT obtained with the pAc373 expression vector was purified as B p66/p60 heterodimer. The recombinant His‐RT was expressed with the pBlueBacHis vector. Thereby, the protein was tagged with an N‐terminal hexahistidine peptide and it was purified as a p70/p70 homodimer. The two enzymes differed in their specific activity, kinetic properties, and in vitro activation by viral and non‐viral proteases. The recombinant His‐RT exhibited a lower specific activity than the recombinant RT. The latter yielded enzyme activities as high as an Escherichia coli ‐expressed RT. Removal of the hexahistidine tag from the recombinant His‐RT by digestion with enterokinase resulted in a complete loss of enzyme activity. Thus, the hexahistidine tag might be an intrinsic part of the active recombinant His‐RT.
Mouse Preproendothelin‐1 Gene
Authors: Chan Theobald Sing Kwok , Lin Colin Xiao Feng , Chan Wood Yee , Chung Stephen Sum Man , Chung Sookja Kim
Published: December 2004

Endothelin‐1 (ET‐1) is a peptide implicated in a wide variety of functions involving vascular and non‐vascular systems. We have cloned the cDNA encoding the mouse preproendothelin‐1 (PPET‐1) and determined its nucleotide sequence. The putative PPET‐1 peptide processing sites are, all conserved and the deduced 21‐amino‐acid mature ET‐1 peptide is identical to that of the rat, human, bovine, porcine and rabbit. Using the cloned cDNA as a probe for in suit hybridization, we detected PPET‐1 mRNA in different tissues at different stages of mouse embryonic development. Embryos at a stage as early as 9.5 days postcoitum (E9.5) have very strong expression in the branchial epithelium, optic vesicle and the endothelial cells of large blood vessels, including the dorsal aorta and aortic arches. While the expression level in the branchial epithelium was decreasing towards the later stage of embryogenesis, the expression in the endothelial cells increased with age. At E10.5, PPET‐1 mRNA was also detected in the otic vesicle as well as in the developing gut epithelium. At later stage of development, the expression of PPET‐1 was primarily found in the vascular endothelial cells, cochlea, eye and the gut, with the highest level of PPET‐I mRNA in the endothelial cells of the lung. These data will be useful for analyzing the function of ET‐1 in these organs.
NMR Studies and Restrained‐Molecular‐Dynamics Calculations of a Long A+T‐Rich Stretch in DNA
Authors: Leijon Mikael , Zdunek Janusz , Fritzsche Hartmut , Sklenar Heinz , Gräslund Astrid
Published: December 2004

The nonamer duplex d(GCAAAAACG) · d(CGTTTTTGC) was studied by 1H‐NMR at 500 MHz. With the exception of the H5′ and H5″ sugar protons, all protons were assigned by two‐dimensional NMR experiments [NOE spectroscopy (NOESY), double‐quantum‐filtered J ‐correlated spectroscopy (DQF‐COSY) and total correlation spectroscopy (TOCSY)]. The exchange kinetics of the imino protons of the Watson‐Crick base pairing were studied it 15°C by measuring inversion‐recovery rates under conditions of extensive ammonia base catalysis. Extrapolation to infinite base concentration gave anomalous long lifetimes for the A‐tract in accordance with previous results [Leroy, J.‐L., Charettier, E., Kochoyan, M. & Guéron, M. (1988) Biochemistry 27, 8894–8898]. On average, 11 NOESY distance constraints/nucleotide were evaluated using the complete relaxation matrix approach. Deoxyribose coupling constants were obtained from simulations of the DQF‐COSY cross‐peaks, assuming a rapid two‐slate equilibrium between a C2′‐endo and C3′‐endo conformer. The sugars were found to be predominantly in the C2′‐endo conformation. The NMR‐derived distance and torsion constraints were implemented into three different restrained‐molecular‐dynamics (rMD) protocols, two in vacuo, with different charges on the phosphate group and the third with the solvent explicitly included. All protocols displayed good convergence from different starting structures. The structures derived from the three protocols satisfied experimental restraints equally well and had similar final energies. Although the overall pattern of sequence dependence of helical parameters shows smile resemblance in all structures, we field that the absolute amplitudes of the parameters are largely dependent on the rMD protocols, particularly the twist parameters. The minor groove distance P (n+2)–P (m+2) varies from 0.7 nm to 1.2 nm in the three protocols. Still the NOESY‐derived anomalously short distances AH2(n)‐H1′(m+1) and AH2(n)‐H1′(n+1), n and m denote complementary residues, which are assumed to he indicative of a compressed minor groove, are kept in all calculated structures.
Identification of mRNAs that Show Modulated Expression During Colon Carcinoma Cell Differentiation
Authors: Van Belzen Nico , Diesveld Michaël P. G. , Van Der Made Angelique C. J. , Nozawa Yoshihiro , Dinjens Winand N. M. , Vlietstra Remko , Trapman Jan , Bosman Fred T.
Published: December 2004

To investigate the working hypotheses that stem cells or their early descendants are prime targets for neoplastic transformation, and that the degree to which a neoplasm retains the immature phenotype is an important determinant of tumor aggressiveness, we have identified several mRNAs that are downregulated during the in vitro differentiation of HT29‐D4 colon carcinoma cells. These genes include heat‐shock cognate protein Hsc70, adenylosuccinate lyase, B23/nucleophosmin, α‐tubulin, and a novel gene designated DS‐1. The Ds‐1 mRNA has a length of approximately 0.9 kb and is downregulated 4.7‐fold upon differentiation. From the DS‐1 cDNA, a protein of 206 amino acids with a molecular mass of 24 kDa and an isoelectric point of 10.9 can be deduced. An antiserum directed against a synthetic peptide detected a minor band of the expected size in Western blots, its well as a major band of lower size that may represent a processed form of the protein.
Effects of Poliovirus 2Apro on Vaccinia Virus Gene Expression
Authors: Feduchi Elena , Aldabe Rafael , Novoa Isabel , Carrasco Luis
Published: December 2004

The effects of transient expression of poliovirus 2Apro on p220 cleavage in COS cells have been analyzed. When 2Apro was cloned in plasmid pTM1 and transiently expressed in COS cells, efficient cleavage of p220 occurred after infection of these cells with a recombinant vaccinia virus hearing phage T7 RNA polymerase. High numbers of COS cells were transfected with pTM1‐2A, as judged by p220 cleavage, thereby allowing an analysis of the effects of poliovirus 2Apro on vaccinia virus gene expression. A 40–50% cleavage of p220 by transfected poliovirus 2Rpro was observed ten hours post infection and cleavage was almost complete (80–90%) 20–25 hours post infection with vaccinia virus, Profound inhibition of vaccinia virus protein synthesis was detectable ten hours post infection and was maximal 20–25 hours post infection. This inhibition resulted from neither a blockade of transcription of vaccinia virus nor a lack of translatability of the mRNAs present in cells that synthesize poliovirus 2Apro. Addition of ara‐C inhibited the replication of vaccinia virus and allowed the continued synthesis of cellular proteins. Under these conditions, 2Apro is expressed and blocks cellular translation. Finally, p220 cleavage by 2Apro did not inhibit the translation of a mRNA encoding poliovirus protein 2C, as directed by the 5′ leader sequences of encephalomiocarditis virus. Therefore, these findings show a correlation between p220 cleavage and inhibition of translation from newly made mRNAs. Our results are discussed in the light of present knowledge of p220 function, and new approaches are considered that might provide further insights into the function(s) of initiation factor eIF‐4F.
Synthesis and Characterization of a Monomeric Mutant Cu/Zn Superoxide Dismutase with Partially Reconstituted Enzymic Activity
Authors: Banci Lucia , Bertini Ivano , Chiu Chi Ying , Mullenbach Guy T. , Viezzoli Maria Silvia
Published: December 2004

A monomeric analog of human Cu/Zn superoxide dismutase (F50E/G51E SOD), previously characterized and found to have reduced enzymic activity, was here further modified by replacing Glu133 with Gln. This substitution does not dramatically affect the coordination geometry at the active site, but enhances enzymic activity, and also increases the affinity for anions at the active site. This behavior parallels earlier published results in which this point mutation was made in the dimeric wild‐type enzyme. The analog described here has afforded for the first time a monomeric superoxide dismutase with substantial activity. This point mutation does not significantly influence the protein structure but interactions with anions, including superoxide, are altered with respect to the monomeric form. The present monomeric Glu133Gln mutant has partially restored enzymic activity. The diminished activity of the monomeric analogs is discussed in the light of possible minor structural changes and some of their characteristics are compared with those of naturally occurring mutants associated with various neurological diseases.
Characterization of a Life‐Cycle‐Stage‐Regulated Membrane Protein Tyrosine Phosphatase in Trypanosoma Brucei
Authors: Bakalara Norbert , Seyfang Andreas , Davis Charles , Baltz Théo
Published: December 2004

We report the first characterization of plasma‐membrane‐bound tyrosine phosphatase activity in the haemoprotozoan, Trypanosoma brucei. Several enzymic properties of the membrane fraction were identical to other protein tyrosine phosphatases (PTPases), such as (a) insensitivity to inhibitors of other protein phosphatases, including tetramisole, sodium tartrate and okadaic acid, (b) inhibition by sodium vanadate, and (c) activation by spermidine. Additionally, T. brucei PTPase activity presented two novel features, an acidic pH optimum at pH 4.0–5.0 and a very low Km value (2.5 nM) for the specific synthetic substrate, Tyr(P)Raytide. Higher Km values of 170 nM for Tyr(P)‐RCML, (RCML, reduced, carboxamidomethylated and maleylated lysozyme) and of 3 mM for the non‐specific inorganic substrate p‐nitrophenyl phosphate, suggested that the PTPase activity of T. brucei was substrate specific. Reconstitution experiments on bloodstream‐stage membrane proteins revealed that three polypeptides of 148, 115 and 72 kDa contained vanadate‐inhibitable PTPase activity. Modulator assays revealed that the 72‐kDa protein was responsible for the observed spermidine stimulation, but indicated that the modulator profile of the 148‐kDa protein was most similar to the whole membrane fraction. Furthermore, the PTPase activity of T. brucei was life‐cycle‐stage regulated. Neither the whole membrane fraction nor the reconstituted proteins of the procyclic insect stage dephosphorylated tyrosine residues.
Analysis of the Structural Organization and Thermal Stability of two Spermadhesins
Authors: Menéndez Margarita , Gasset María , Laynez José , López‐Zumel Consuelo , Usobiaga Pilar , Töpper‐Petersen Edda , Calvete Juan J.
Published: December 2004

The CUB domain is a widespread 110‐amino‐acid module found in functionally diverse, often developmentally regulated proteins, for which an antiparallel β‐barrel topology similar to that in immunoglobulin V domains has been predicted. Spermadhesins have been proposed as a subgroup of this protein family built up by a single CUB domain architecture. To test the proposed structural model, we have analyzed the structural organzation of two members of the spermadhesin protein family, porcine seminal plasma proteins I/II (PSP‐I/PSP‐TI) heterodimer and bovine acidic seminal fluid protein (aSFP) homodimer, using differential scanning calorimetry, far‐ultraviolet circular dichroism and Fourier‐transform infrared spectroscopy. Thermal unfolding of PSP‐I/PSP‐II and aSFP were irreversible and followed a one‐step process with transition temperatures (Tm) of 60.5°C and 78.6°C, respectively. The calorimetric enthalpy changes (ΔHcal) of thermal denaturation were 439 kJ/mol for PSP‐I/PSP‐II and 660 kJ/mol for aSFP dimer. Analysis of the calorimetric curves of PSP‐I/PSP‐II showed that the entire dimer constituted the cooperative unfolding unit. Fourier‐transform infrared spectroscopy and deconvolution of circular dichroic spectra using a convex constraint analysis indicated that β‐structure and turns are the major structural element of both PSP‐I/PSP‐II (53% of β‐sheet, 21% of turns) and aSFP (44% of β‐sheet, 36% of turns), and that the porcine and the bovine proteins contain little, if any, α‐helical structure. Taken together, Our results indicate that the porcine and the bovine spermadhesin molecules are probably all‐β‐structure proteins, and would support a β‐barrel topology like that predicted for the CUB domain. Other β‐structure folds, such as the Greek‐key pattern characteristic of many carbohydrate‐binding protein domains cannot be eliminated. Finally, the same combination of biophysical techniques was used to characterize the residual secondary structure of thermally denatured forms of PSP‐I/PSP‐II and aSFP, and to emphasize the aggregation tendency of these forms.
Recognition of tRNAPhe by Phenylalanyl‐tRNA Synthetase of Thermus Thermophilus
Authors: Moor Nina A. , Ankilova Valentina N. , Lavrik Olga I.
Published: December 2004

The tRNAPhe nucleotides required for recognition by phenylalanyl‐tRNA synthetase of Thermus thermophilus have been determined using Escherichia coli tRNAPhe transcripts with various mutations. The anticodon nucleotides are shown to be the most important recognition elements. The discriminator nucleotide, A73, involved in the recognition set of yeast, E. coli and human phenylalanyl‐tRNA synthetases contributes only slightly to tRNAPhe recognition by Th. thermophilus phenylalanyl‐tRNA synthetase. Nucleotide 20 and some tertiary nucleotides, including the conserved G19 · C56 base pair, are proposed to participate in stabilization of the precise tRNA conformation required for efficient aminoacylation. The role of the 3′‐CCA terminus, common to all tRNAs, in the specific interaction of tRNA with phenylalanyl‐tRNA synthetase is discussed.
Conidial dispersal of Gremmeniella abietina: climatic and microclimatic factors
Authors: Petäistö R.‐L. , Heinonen J.
Published: December 2004

The conidia dispersal started in Suonenjoki, in central Finland, in 1997–99 by the end of May or beginning of June, and continued occasionally at least to the middle of September. The temperature sum, day degrees (d.d., threshold temperature = 5°C) was between 100 and 165 d.d. at the beginning of dispersal. In years 1997–99, 80, 94 and 82% of the dispersal had occurred by the end of July – beginning of August when the temperature sum reached 800 d.d. All the spore data are coming from the spore traps. Cumulative number of conidia increased linearily with logarithm of temperature sum. A binary logistic regression model with temperature sum and rainfall as explanatory variables predicted accurately the date of the first spores in the spring: the prediction error was at most 3 days. The model classified 69% of all the days in the analysis correctly to the spore‐free days and correspondingly 74% to the days of at least one spore caught. A regression model for the number of spores per day explains 21, 5 and 51% of the within‐season variation in 1997–99 (24, 37 and 62% on a logarithmic scale). The explanatory weather variables in the model were d.d., rain and year. The very low explanatory coefficient of determination in 1998 results from one exceptionally high number of conidia. The between‐differences in the total number of spores were large and could not be explained by the measured weather variables. In the regression model, these differences were taken into account by adding a constant for each year in the model. Rain increased conidia dispersal significantly but conidia were found also in consecutive rainless days.
Glomerulosclerosis: A paraneoplastic phenomenon?
Authors: LOI SHERENE , PERRY GREGORY J , STANDISH HUGO , DOWLING JOHN
Published: December 2004

SUMMARY:  Glomerulosclerosis is not classically considered a paraneoplastic glomerular lesion. Focal and segmental glomerulosclerosis (FSGS) has rarely been reported in association with solid tumours. We report three cases of FSGS and an additional case of collapsing glomerulosclerosis in patients presenting with nephrotic syndrome and malignancy.
Periodic limb movement disorder and restless legs syndrome in dialysis patients
Authors: RIJSMAN ROSELYNE M , DE WEERD AL W , STAM CJ , KERKHOF GRARD A , ROSMAN JOHAN B
Published: December 2004

Background:  Sleep disturbances, in particular restless legs or limb movements, during the night are often reported by uremic patients. However, polysomnography (PSG) studies have never been carried out to confirm the actual occurrence of these disorders and the association with other objective and self‐reported sleep–wake data.

Methods:  Forty‐eight participants were subjected to a 2‐day PSG. These data on sleep including periodic limb movements, which are associated with restless legs, were correlated with clinical observations, quality of sleep–wake and life questionnaires, and with biochemical and neurographical measures.

Results:  Restless legs syndrome (RLS) was observed in 58.3% of the patients and periodic limb movement disorder (PLMD) occurred in 70.8% of the patients. PLMD was revealed polysomnographically in almost 90% of the RLS patients. Patients with both PLMD and RLS had significantly poorer sleep quality than those with neither disorder or with PLMD alone, both in terms of self‐reported data and the PSG. Quality of life was significantly worse in patients with RLS and PLMD compared to those patients with neither disorder. PLMD patients also tended to have a lower quality of life. All other metabolic  measures  and  the  results  of  a  nerve  conduction  test  were  not  correlated  with  RLS  and/or PLMD.

Conclusion:  There was a high prevalence of severe RLS and PLMD in the present sample of uraemia patients. Nearly all RLS patients had severe PLMD. RLS (in combination with PLMD) in dialysis is associated with poor sleep quality, insomnia complaints, depression and emotional distress. Our results suggest that PLMD per se is also clinically relevant.
Ultrasonography in the management of exit site infections in peritoneal dialysis patients
Authors: KWAN TZE‐HOI , TONG MATTHEW KA‐HANG , SIU YUI‐PONG , LEUNG KAY‐TAI , LUK SAU‐HAR , CHEUNG YU‐KEUNG
Published: December 2004

Aim:  To assess the efficacy of using ultrasonography (USG) in monitoring the progress of exit site infection (ESI) in patients undergoing continuous ambulatory peritoneal dialysis (CAPD).

Methods:  Twenty‐two cases of newly diagnosed ESI and 20 cases with normal exit sites as controls were assessed by using USG. The exit sites were reassessed by using USG after finishing a course of antibiotic therapy, and the sonographic findings were correlated with the clinical outcome.

Results:  Out of the 22 cases of ESI, 21 cases had definite sonolucent zones around the external cuffs, while one case had normal sonographic findings. Of the 20 control cases of normal exit sites, 16 had normal sonographic findings, and four had sonolucent zones around the external cuffs. Exit site infections correlated with positive sonographic findings as compared to normal exits (P < 0.0001). The 21 cases of ultrasonic‐positive ESI were re‐examined after antibiotic therapy, and 10 of these had a post‐treatment sonolucent rim around the distal cuff ≤1 mm thick, while 11 cases were persistently >1 mm thick. The former group was shown to have a more favourable outcome (P = 0.013). And despite variable USG findings, all eight patients with Pseudomonas aeruginosa‐related ESI had an unfavourable clinical outcome.

Conclusion:  Ultrasonography of the exit sites in CAPD patients is a useful adjunctive tool in the management of ESI. A sonolucent zone around the external cuff >1 mm thick following a course of antibiotic treatment and the involvement of the proximal cuff are associated with poor clinical outcome. In ESI caused by Pseudomonas aeruginosa, the clinical outcome was uniformly poor irrespective of the sonographic findings.
Efficacy and side‐effect profile of sevelamer hydrochloride used in combination with conventional phosphate binders
Authors: STURTEVANT JOANNA M , HAWLEY CARMEL M , REIGER KYLIE , JOHNSON DAVID W , CAMPBELL SCOTT B , BURKE JOHN R , BOFINGER ANDREW , ISBEL NICOLE M
Published: December 2004

Background:  Poor phosphate control is common among patients with end‐stage renal disease. Sevelamer hydrochloride has been demonstrated to be a safe and effective phosphate binder when used as a monotherapy. However, cost limits its usefulness in many countries. Data assessing its effectiveness and safety in combination with conventional phosphate binders are lacking.

Methods:  Dialysis patients meeting the following inclusion criteria participated in this study: (i) hyperphosphataemia >1.8 mmol/L (5.6 mg/dL); and (ii) an inability to tolerate currently available binders. The trial was conducted in three phases each lasting 3 months: (i) an observation phase (patients continued on their regular phosphate binders); (ii) a titration phase (sevelamer was added at a dose of 403 mg three times daily with meals, titrated to a maximum of 1209 mg three times daily); and (iii) a maintenance phase.

Results:  Twenty‐five patients were recruited into the study. Eighteen patients completed all three trial phases. Mean serum phosphate dropped from 2.11 ± 0.06 mmol/L (6.6 ± 0.2 mg/dL) during the observation period to 1.91 ± 0.01 mmol/L (5.9 ± 0.003 mg/dL) during the maintenance phase (P = 0.02). Calcium × phosphate product fell from 5.49 ± 0.17 mmol2/L2 (68.64 ± 2.11 mg2 dL2) to 4.89 ± 0.27 mmol2/L2 (61.36 ± 3.35 mg2 dL2) (P = 0.02). There was no significant change in serum calcium or parathyroid hormone. Total serum cholesterol fell from 3.8 mmol/L (3.4–4.37) 147 mg/dL (131–169) to 3.55 mmol/L (2.97–4.2) 137 mg/dL (115–162) (P = 0.02). Serum low‐density lipoprotein cholesterol also fell significantly from 1.67 ± 0.10 mmol/L (65 ± 4 mg/dL) to 1.52 ± 0.11 mmol/L (59 ± 4 mg/dL) (P = 0.04). The average prescribed dose of sevelamer was 2.4 g/day. Elemental calcium dropped from 3.4 g/day (1.4 to 4.6) to 1.2 g/day (0.6–2.4) (P = 0.04). Seventy‐two per cent of patients reported mild flatulence, nausea and indigestion. Three patients discontinued treatment because of adverse effects.

Conclusions:  Sevelamer in combination with conventional phosphate binders is effective in lowering serum phosphate and calcium‐phosphate product in patients with refractory hyperphosphataemia. Beneficial effects on lipid profile were also observed. Mild gastrointestinal upset is common.
From the bush to the beach: water safety in rural and remote New South Wales
Authors: Mitchell Rebecca , Haddrill Katrina
Published: December 2004

Objective:  To describe the types of aquatic locations attended by residents of rural and remote New South Wales (NSW), to record self‐reported water safety‐related behaviour, and identify preferred communication mediums for water safety messages.

Methods:  A stratified random telephone survey was conducted of 500 NSW residents aged greater than 15 years residing in moderately accessible, remote and very remote locations in NSW.

Results:  Results indicate that around two‐thirds of respondents had been in or on the water at a pool, beach, lake, river or dam in the past 6 months. The most common type of aquatic facilities used were rivers, creeks or steams (53.5%), beaches (45.7%), public pools (45.5%), private pools (40.7%), dams (40.6%) and lakes (27.0%). Time spent at each of these locations and time of day each location was visited varied. Overall, the majority of respondents reported practicing water‐related safe behaviour. Preferred communication mediums for water safety messages included television, schools and newspapers.

Conclusions:  Water safety education, especially in relation to beach conditions, remains just as important a topic for public health authorities and key water safety agencies in regional and remote NSW as it is in coastal suburbs. Responses from the survey, along with key stakeholder advice, will be used to inform the development of appropriate strategies aimed to reduce drowning deaths in rural and remote locations in NSW.
Complementary findings in clinical and epidemiologic features of mumps and mumps meningoencephalitis in children without mumps vaccination
Authors: Kanra Güler , Isik Pinar , Kara Ates , Cengiz Ali Bülent , Seçmeer Gülten , Ceyhan Mehmet
Published: December 2004

Background
: Mumps vaccine has not yet been included in the routine vaccination programme, for this reason mumps is still one of the most common infections for children in Turkey. One of the major complication of mumps is meningoencephalitis, which although usually heals spontaneously, it may cause neurologic complications.

Methods
: This study was undertaken to investigate epidemiologic and demographic characteristics in children with mumps and mumps meningoencephalitis and clinical/laboratory findings in children with mumps meningoencephalitis diagnosed over a 11 year period. A total of 2422 mumps and 135 mumps meningoencephalitis cases were covered in this study which constitutes one of the largest series of mumps meningoencephalitis in the literature. The mean age of mumps and mumps meningoencephalitis cases were 6.6 ± 2.7 and 7.6 ± 2.6 years, respectively.

Results
: There was a male predominance both among the cases of mumps and mumps meningoencephalitis. The age and seasonal distributions were similar in the mumps and mumps meningoencephalitis groups. The most common symptoms of mumps meningoencephalitis were fever (97%), vomiting (94%) and headache (88.8%). The mean cerebrospinal fluid (CSF) total cell count and lymphocyte count were 540 ± 460/mm3 and 300 ± 330/mm3, respectively. The mean CSF protein and glucose levels were found to be 56.97 ± 27.94 mg/dL and 53.67 ± 15.46 mg/dL, respectively. The mean of CSF/blood glucose ratio was 0.53 ± 0.16. The mean duration of hospitalization in mumps meningoencephalitis cases was found to be 5.1 ± 2.4 days and this was longer in boys (P < 0.05).

Conclusion
: In patients with mumps meningoencephalitis, higher CSF protein levels and lower CSF glucose/blood glucose ratio were associated with longer hospitalization periods. There was no death.
Dosage regimen of arbekacin for methicillin‐resistant Staphylococcus aureus infection in newborns and infants
Authors: Yoshida Koichi , Minowa Hideki , Ebisu Reiko , Uchida Yumiko , Takahashi Yukihiro , Yoshioka Akira
Published: December 2004

Background
: Arbekacin (ABK) is an aminoglycoside antibiotic that has a dose‐dependent bactericidal action. Because it inhibits the production of toxic shock syndrome toxin‐1 (TSST‐1) by methicillin‐resistant Staphylococcus aureus (MRSA), it has attracted attention as a therapeutic drug for MRSA infection. In this study, the authors investigated the pharmacokinetics of ABK based on therapeutic drug monitoring (TDM), in order to establish an effective dosage regimen with minimal adverse reactions in MRSA infected newborns and infants.

Methods
: Arbekacin was administered to nine MRSA infected newborns and infants between October 2000 and March 2002. Following the initial ABK administration, blood was collected and the blood concentration of ABK was measured. The blood concentrations of ABK were analyzed by the two‐compartment model or by a model‐independent method in order to elucidate the pharmacokinetics of ABK. Pharmacokinetic analysis was performed using WinNonlin Professional V3.1.

Results
: The mean age at initial ABK administration was 24.0 ± 26.0 days (postconceptional age: 39.2 ± 3.9 weeks). The increase in the peak blood concentration of ABK was 2.40 ± 0.20 µg/mL per mg ABK per kg bodyweight, showing great consistency among cases. The elimination half‐life of ABK was 0.22–3.52 h in the α phase (T1/2α) and 2.42–33.44 h in the β phase (T1/2β), showing great variation among cases. The distribution volume was 0.75 ± 0.13 L/kg, and systemic clearance was 0.054 ± 0.012 L/h/kg. ABK alleviated clinical symptoms and inflammations in all cases.

Conclusion
: Nine newborns and infants with MRSA infection and various underlying diseases were successfully treated with TDM‐based administration of ABK with no severe adverse reactions.
Environmental mercury exposure in children: South China's experience
Authors: Ip Patrick , Wong Virginia , Ho Marco , Lee Joseph , Wong Wilfred
Published: December 2004

Background
: Environmental mercury levels significantly increased in the past decades following its increase in industrial applications. In spite of an increasing concern on the potential harmful effects of mercury on children, there is no reported data for the Chinese population. The relationship between dietary habit and environmental mercury exposure in Chinese children was studied.

Methods
: The hair and blood mercury levels of Chinese children aged above 3 years in 2000 March to September, were studied. Sociodemographic data, dietary habits of the past 6 months, and other risk factors for environmental mercury exposure were collected. Those children with blood mercury levels above the toxic range (i.e. > 45 nmol/L) and their family members were further evaluated and their blood and hair mercury levels were monitored before and after Fishing‐Moratorium period (June to August 2000) in South China Sea.

Results
: Altogether, 137 Chinese children (mean age, 7.2 years) were recruited. The mean hair mercury level was 2.2 p.p.m and the mean blood mercury level was 17.6 nmol/L. There was a strong correlation (r = 0.88) between hair and blood mercury levels in our cohort. Frequency of fish consumption correlated with hair (r = 0.51) and blood (r = 0.54) mercury levels. For those children who consumed fish more than 3 times/week, hair and blood mercury levels were twice as high as those who consumed fish l–3 times/week and threefold of those who never consumed fish. Five children and 12 family members had toxic blood mercury levels. Their blood (P < 0.0001) and hair (P = 0.02) mercury levels dropped significantly after reducing fish consumption during Fishing‐Moratorium period.

Conclusion
: Both blood and hair (i.e. Tissue) mercury levels of children in Hong Kong was elevated and correlated with the frequency of fish consumption.
Improved diabetes control by using ‘close adjustment algorithms’
Authors: Miyako Kenichi , Kuromaru Ryuichi , Kohno Hitoshi , Hara Toshiro
Published: December 2004

Background
: Intensive insulin therapy increases the frequency of severe hypoglycemia despite markedly improved glycemic control in patients with type 1 diabetes mellitus. To determine the optimal dose of insulin, the authors designed algorithms based on self‐monitored blood glucose levels.

Methods
: Each dose of insulin was composed of two components: a basal dose determined on the basis of blood glucose levels over the previous two days and an additional dose determined on the basis of blood glucose level just before insulin injection. The patients were instructed to adjust each dose according to the algorithms. The authors investigated the effects of using algorithms on glycemic control, anthropometric data, body composition, and lipid profile in seven females with type 1 diabetes 12–20 years old.

Results
: After 3 months, the daily dose of insulin increased significantly from 0.93 ± 0.18 to 1.16 ± 0.26 units/kg of body weight, and haemoglobin A1C decreased significantly from 8.27 ± 1.33 to 6.50 ± 0.64%. Severe hypoglycemia, however, did not occur. Body mass index increased significantly from 21.7 ± 2.7 to 22.7 ± 2.9 kg/m2 with no increase in the percentage of body fat. All lipid‐profile data showed a decreasing trend.

Conclusions
: Algorithms developed on the basis of self‐monitored blood glucose levels are useful in determining the optimal dose of insulin and can improve glycemic control and lipid metabolism.
Leptin and the metabolic syndrome in Korean adolescents: Factor analysis
Authors: Park Hye Soon , Lee Myoung Sook , Park Jung Yul
Published: December 2004

Background
: The objectives of this study were to evaluate the relationships of leptin with the metabolic syndrome and to examine leptin's role in clustering of the metabolic components among Korean adolescents.

Methods
: A cross sectional study was carried out in 68 male and 80 female adolescents aged 13−18 years in an urban area of South Korea. Anthropometric variables were measured and blood pressure, fasting plasma glucose, low‐density lipoprotein cholesterol, triglyceride, high‐density lipoprotein cholesterol, and serum leptin were obtained.

Results
: As more metabolic components were clustered, body mass index, body fat, waist circumference, and serum leptin levels were significantly increased. Principal components factor analysis revealed three factors in males and females that explained 70% and 65%, respectively, of the observed variance of the 10 measured variables. These were obesity‐leptin‐lipid factor, blood pressure factor, and glucose‐cholesterol factor in males and obesity‐leptin‐glucose factor, blood pressure factor, and cholesterol factor in females. Leptin loaded on only one factor in both genders.

Conclusion
: Leptin did not appear to have a major role linking various components of the metabolic syndrome, even though it was strongly associated with obesity indices. Gender difference of linking of leptin with glucose or lipid was observed. There seems to be more than one pathophysiological mechanism which might underlie full expression of the metabolic syndrome among Korean adolescents.
Clinical application of prognostic factors for patients with organic causes of erectile dysfunction on 100 mg of sildenafil citrate
Authors: CHIA SING JOO , RAMESH KAMALAKANNAN , EARNEST ARUL
Published: December 2004

Abstract  Objectives  There are still no clinical predictive factors to determine the response rate of erectile dysfunction (ED) patients to sildenefil citrate. The aim of the present study is to evaluate and stratify the risk factors and attempted to determine the prognostic factors in clinical practice to predict the response rate. This is important in improving cost effectiveness and avoiding side‐effects.

Material and methods  This is an open label prospective study including patients attending the andrology clinic in Tan Tock Seng Hospital, Singapore, over 2 years. The patients were evaluated and investigated for possible underlying causes and ED severity was assessed by five‐items of the International Index of Erectile Function questionnaire (IIEF‐5), together with the duration, degree and rigidity of erection. Psychogenic causes were excluded with a minimal follow up of 6 months. All patients were placed on 100 mg of sildenafil citrate and were reassessed at the end of 6 months. Logistic regression with univariate and multivariate was used as the method of statistical analysis.

Results  A total of 232 patients were in the cohort. The overall response rate was 43%, with the best response rate in veno‐occlusive cases and the worst responses from neurogenic causes. Age, smoking, diabetes mellitus, hypertension, hyperlipidemia, pretreatment IIEF‐5 score, interval to achieve erection and duration of erection were significant in univariate analysis, but only age, smoking and IIEF‐5 score were significant in multivariate analysis. With a combination of these factors, a table was formed to determine the possible response rate in clinical practice. This will assist physicians in selecting patients with potentially favorable responses and avoid side‐effects and an unnecessary wastage of time and cost.

Conclusion  Possible factors could be determined and used clinically to predict the response rate to sildenafil citrate.
Three‐dimensional ultrasonography in evaluation of benign prostatic hyperplasia
Authors: KANAO KENT , KIKUCHI EIJI , NAKASHIMA JUN , HORIGUCHI YUTAKA , NAKAGAWA KEN , OYA MOTOTSUGU , OHIGASHI TAKASHI , MARUMO KEN , MURAI MASARU
Published: December 2004

Abstract  Background  The clinical application of volume estimation by 3‐D ultrasound has recently gained much attention. However, there have been no reports evaluating the prostate volume by 3‐D transrectal ultrasound (TRUS) before transurethral resection of the prostate (TURP). The purpose of the present study was to evaluate the value of 3‐D TRUS for prediction of prostate morphology and resected weight before TURP and to investigate whether 3‐D TRUS is a more useful examination than 2D TRUS in patients with benign prostatic hyperplasia (BPH).

Methods  Transurethral resection of the prostate was performed in 23 patients with BPH. We evaluated the prostate morphology and measured both the volumes of the whole prostate and the transition zone using 2‐D and 3‐D TRUS, respectively. The actual resected weight was recorded and compared with the volume of the whole prostate and that of the transition zone measured by 2D and 3‐D TRUS.

Results  The volume of the transition zone measured by 3‐D TRUS correlated most strongly with the resected weight (r = 0.84). A large median lobe was seen in three patients in whom the transition zone volume measured by 2‐D TRUS was considerably larger than the resected weight. However, overestimation in the three patients decreased by the use of 3‐D TRUS.

Conclusions  It was concluded that 3‐D TRUS was equal or superior to 2‐D TRUS in prediction of the resected weight and 3‐D TRUS offers better information as a diagnostic tool before TURP.
Congenital absence of the testis in human fetuses and in cryptorchid patients
Authors: FAVORITO LUCIANO A , KLOJDA CARLOS AB , SAMPAIO FRANCISCO JB
Published: December 2004

Abstract  Background  The aim of the present study is to make a comparative study in human fetuses and in patients with cryptorchidism, analyzing the incidence of a number anomalies of the testes for both populations.

Methods  We studied 326 testes from 163 human fetuses ranging in age from 10 to 35 weeks postconception (WPC) and 133 testes from 101 cryptorchid patients aged from 1 to 15 years old (mean, 6.4 years). The Fisher's exact test was used for comparison.

Results  Among 326 fetal testes, 224 (68.7%) were abdominal, 45 (13.8%) were inguinal and 55 (16.8%) were scrotal. In one fetus at 23 WPC, both testes (0.6%) were absent. Of the 133 cryptorchid testes, 17 (12.78%) were abdominal, 92 (69.1%) were inguinal and 24 (18%) were high scrotal. Of the 17 abdominal testes, three (17.6%) were atrophic and two were vanished (11.7%). Of the 92 inguinal testes, one (1.08%) was vanished. Twenty‐eight (21%) of the cryptorchid testes were impalpable and among these, 17 were located in the abdomen (60.7%) and 11 (38.2%) in the inguinal region (internal ring).

Conclusions  Testicular agenesis is a very rare anomaly, both in fetuses and patients with cryptorchidism.
Synchronous anal and penile cancers
Authors: WYSOCKI WOJCIECH M , KOMOROWSKI ANDRZEJ L , KOLODZIEJSKI LESZEK S , WYSOCKA JOANNA
Published: December 2004

Abstract  Malignancies of the anogenital region have similar etiology, including viral infections, sexual habits, social and educational factors. Condylomata acuminata represent precancerous lesions. In the available literature, we found no cases of synchronous cancers of anus and penis reported. We report a case of synchronous cancer of the penile foreskin and cancer of the anus in a 36 year‐old man. The microscopic diagnosis of the anal lesion was squamous cell cancer, basaloid type. The penile lesion was squamous cell cancer, keratinizing type. Both cancers presented with the in situ component. Immunohistochemical staining (MIB‐1) and histological features (koilocytosis, acanthosis and dyskeratosis) indirectly confirmed the past viral infection. Numerous etiological similarities in anogenital cancers entail careful clinical inspection in patients with a malignant lesion in this region and suggest searching for possible other concomitant cancers or precancerous lesions.
Telomerase activity is correlated with lower grade and lower stage bladder carcinomas
Authors: OKUMURA AKIOU , MIZUNO ICHIRO , NAGAKAWA OSAMU , FUSE HIDEKI
Published: December 2004

Abstract  Background:  Telomerase is a ribonucleoprotein enzyme that compensates for the progressive erosion of telomeres. The increasing interest in telomerase is motivated by the demonstration that most human carcinomas are telomerase positive. The potential use of telomerase activity in bladder carcinomas using a urine sample has been reported in several studies. However, little is known about the detection of telomerase activity in bladder carcinoma tissues. Herein, we investigate telomerase activity in bladder carcinoma tissues according to grade (G) and stage.

Material and methods:  Telomerase activity was assayed by polymerase chain reaction enzyme‐linked immunosorbent assay methods. Malignant lesions were assessed in 37 patients with bladder carcinoma and no malignant lesions were assessed in five patients with dysplasia or inflammatory bladder lesions.

Results:  Twenty‐three out of 37 carcinoma samples were telomerase‐positive and one out of five control samples without carcinoma was telomerase‐positive. The positive rates according to stage and grade were 83.3% for superficial and 42.1% for invasive stages and 83.3% for G1, 66.7% for G2 and 40.0% for G3. Telomerase activity was correlated with lower grade and lower stage bladder carcinomas.

Conclusion:  These results strongly suggest that reactivation of telomerase may differ between superficial and invasive bladder carcinomas and also between low grade and high grade bladder carcinomas.
Transurethral resection in women with lower urinary tract symptoms
Authors: QIU YIQING , DU CHUANJUN , SHEN XIAOCAO
Published: December 2004

Abstract  Background:  To evaluate the preliminary safety, efficacy and complications of transurethral resection (TUR) in the management of women with lower urinary tract symptoms.

Methods:  One hundred and eight women (mean age, 57.6 years) with moderate to severe lower urinary tract symptoms underwent TUR between September 1998 and December 2002. At preoperative baseline, 6 months and 24 months postoperatively, all patients underwent clinical evaluations including the standardized American Urological Association symptom score, peak urine flow, postvoid residual urine volume and quality‐of‐life assessments. Operative time, catheter time, hospital stay and incidence of side‐effects were also recorded.

Results:  The total mean operative time was 19.25 ± 6.70 min. The mean catheter time was 26.2 ± 12.8 h and the mean hospital stay was 2.08 ± 0.84 days. No patient needed a blood transfusion or experienced hyponatremia. The total storage and voiding symptom scores and quality of life score had improved significantly 6 months after the surgical procedure (P < 0.01). There were little changes in peak flow rate and residual urine volume. Complications of the procedure included urinary tract infections (n = 4) and stress incontinence (n = 3). Two patients later underwent bladder neck suspension procedure.

Conclusions:  In this preliminary study, there was significant clinical improvement maintained at all follow‐up intervals and the results were not comparable to TUR of the prosatate in men with clinical benign prostatic hyperplasia. The risks associated with TUR in women, especially of stress incontinence, are significant. A multicenter clinical trial is currently underway to determine the long‐term efficacy and safety of TUR in women.
Median raphe cyst in the scrotum, mimicking a serous borderline tumor, associated with cryptorchidism after orchiopecxy
Authors: HARA NOBORU , KAWAGUCHI MAKOTO , KOIKE HIROSHI , TAKAHASHI KOTA
Published: December 2004

Abstract  Median raphe cyst (MRC) is a benign lesion occurring predominantly in the ventral surface of the penises of young men and is an embryological developmental anomaly of the male genitalia. Serous borderline tumors (SBT) are found most frequently in the female ovary and only several cases with SBT of the male genitalia have been reported. We describe a case of MRC with features of SBT, which appeared in the scrotum of a 9‐year‐old boy after orchiopexy and was associated with surgery for cryptorchidism. The cyst arose on the right testicular tunica and consisted of cystic components with intracystic papillae lined by stratified epithelial cells, some of which showed mild cytological atypia and sporadic mitosis. These epithelial cells expressed CA 125, CA 19‐9, carcinoembryonic antigen, estrogen receptor and progesterone receptor. Although no cases of MRC with characteristics of SBT in association with the rete testis has been described, the current report gives additional information for follow‐up of cryptorchidism.
Effects of agar (kanten) diet on obese patients with impaired glucose tolerance and type 2 diabetes
Authors: Maeda H. , Yamamoto R. , Hirao K. , Tochikubo O.
Published: December 2004

Aim:  The aim of this study was to evaluate the efficacy of agar diet in combination with a conventional diet (traditional Japanese food) for obese patients with impaired glucose tolerance and type 2 diabetes.

Methods:  After a 4‐week run‐in period on their habitual diets, 76 patients were randomly assigned to have conventional diet or conventional diet with agar. Both groups were on these diets for 12 weeks. Body weight, body mass index (BMI), glycaemic control, blood pressure, insulin resistance, total body fat, fat distribution and lipids were assessed before and after the experimental period.

Results:  In both groups, after 12 weeks, mean body weight, BMI, fasting glucose levels, homeostasis model assessment‐insulin resistance, and systolic and diastolic blood pressures had decreased significantly from their baseline values. HbA1c, visceral fat area, subcutaneous fat area, total body fat, insulin area under the curve after oral glucose tolerance test and total cholesterol decreased significantly in the agar‐diet group. After 12 weeks, mean changes of body weight (−2.8 ± 2.7 kg vs. −1.3 ± 2.3 kg, p = 0.008), BMI values (−1.1 ± 1.1 kg/m2 vs. −0.5 ± 0.9 kg/m2, p = 0.009) and total cholesterol (−7.6 ± 27.5 mg/dl vs. + 2.4 ± 23.4 mg/dl, p = 0.036) were significantly greater in the agar‐diet group than in the conventional diet group.

Conclusions:  The agar diet resulted in marked weight loss due to the maintenance of reduced calorie intake and to an improvement in metabolic parameters.