bluuebunny/crossref_metadata_2025 huggingface dataset:
Biochemical characterization of the respiratory syncytial virus P–P and P–N protein complexes and localization of the P protein oligomerization domain
Authors: Castagné Nathalie , Barbier Alexandra , Bernard Julie , Rezaei Human , Huet Jean-Claude , Henry Céline , Costa Bruno Da , Eléouët Jean-François
Published: May 2004

The RNA-dependent RNA polymerase complex of respiratory syncytial virus (RSV) is composed of the large polymerase (L), the phosphoprotein (P), the nucleocapsid protein (N) and the co-factors M2-1 and M2-2. The P protein plays a central role within the replicase–transcriptase machinery, forming homo-oligomers and complexes with N and L. In order to study P–P and N–P complexes, and the role of P phosphorylation in these interactions, the human RSV P and N proteins were expressed in E. coli as His-tagged or GST-fusion proteins. The non-phosphorylated status of recombinant P protein was established by mass spectrometry. GST-P and GST-N fusion proteins were able to interact with RSV proteins extracted from infected cells in a GST pull-down assay. When co-expressed in bacteria, GST-P and His-P were co-purified by glutathione-Sepharose affinity, showing that the RSV P protein can form oligomers within bacteria. This result was confirmed by chemical cross-linking experiments and gel filtration studies. The P oligomerization domain was investigated by a GST pull-down assay using a series of P deletion constructs. This domain was mapped to a small region situated in the central part of P (aa 120–150), which localized in a computer-predicted coiled-coil domain. When co-expressed in bacteria, RSV N and P proteins formed a soluble complex that prevented non-specific binding of N to bacterial RNA. Therefore, RSV P protein phosphorylation is not required for the formation of P–P and N–P complexes, and P controls the RNA binding activity of N.
p16INK4A-independence of Epstein–Barr virus-induced cell proliferation and virus latency
Authors: Hayes Michelle J. , Koundouris Anna , Gruis Nelleke , Bergman Wilma , Peters Gordon G. , Sinclair Alison J.
Published: May 2004

Epstein–Barr virus (EBV) has the ability to promote cell cycle progression following the initial infection of primary resting B-lymphocytes and to cause cell cycle arrest at the onset of the viral replicative cycle. Various mechanisms have been proposed for the proliferative effects, including the up-regulation of cyclin D2 by the viral EBNA-2 and EBNA-LP proteins, direct binding of EBNA3C to the retinoblastoma protein (pRb), and down-regulation of the p16INK4A tumour suppressor by the viral LMP1 product. To try to gain insight into the relative importance of these mechanisms, the ability of EBV to immortalize lymphocytes from an individual who is genetically deficient for p16INK4A was examined. From detailed analyses of the resultant lymphoblastoid cell lines it is concluded that p16INK4A status has little bearing on EBV's ability to manipulate the cell cycle machinery and a model to accommodate the previously proposed routes taken by EBV to bypass the restriction point is presented.
Identification of membrane proteins differentially expressed in human papillomavirus type 16 E5-transfected human keratinocytes by nanoelectrospray ionization mass spectrometry
Authors: Leykauf Kerstin , Salek Mojiborahman , Schlüter Holger , Lehmann Wolf-Dieter , Alonso Angel
Published: May 2004

Membrane proteins differentially expressed in human papillomavirus type 16 (HPV-16) E5-transfected HaCaT cells have been identified. Membrane proteins were isolated and separated by two-dimensional gel electrophoresis. Spots showing quantitative differences between E5-transfected and control cells were extracted and the proteins were identified by nanoelectrospray ionization mass spectrometry. A total of 24 spots was analysed. Among the proteins showing differential expression, a decreased amount of calnexin and increased expression of hsp70, proteins both involved in maturation and transport of MHC class I complexes to the plasma membrane, were noticed. These findings correlate with the decreased surface expression of MHC class I molecules described in E5-expressing cells, HPV-positive cervical lesions and cervical carcinomas. These results stress the value of the proteomic approach, as used here in the experimental design, which allows the correlation of changes in host gene expression with biological functions of viral genes.
During entry of alphaviruses, the E1 glycoprotein molecules probably form two separate populations that generate either a fusion pore or ion-permeable pores
Authors: Wengler Gerd , Koschinski Andreas , Wengler Gisela , Repp Holger
Published: May 2004

Studies using the alphavirus Semliki Forest virus have indicated that the viral E1 fusion protein forms two types of pore: fusion pores and ion-permeable pores. The formation of ion-permeable pores has not been generally accepted, partly because it was not evident how the protein might form these different pores. Here it is proposed that the choice of the target membrane determines whether a fusion pore or ion-permeable pores are formed. The fusion protein is activated in the endosome and for steric reasons only a fraction of the activated molecules can interact with the endosomal membrane. This target membrane reaction forms the fusion pore. It is proposed that the rest of the activated molecules interact with the membrane in which the protein is anchored and that this self-membrane reaction leads to formation of ion-permeable pores, which can be detected in the target membrane after fusion of the viral membrane into the target membrane.
Multiple human immunodeficiency virus type 1 Nef functions contribute to efficient replication in primary human macrophages
Authors: Brown Amanda , Moghaddam Shaghayegh , Kawano Thomas , Cheng-Mayer Cecilia
Published: May 2004

The human immunodeficiency virus type 1 (HIV-1) Nef protein has been shown to accelerate viral growth kinetics in primary human T-lymphocytes and macrophages; however, the specific function(s) of Nef responsible for this phenotype in macrophages is unknown. To address this issue, mutants of a molecularly cloned macrophage-tropic isolate, HIV-1SF162, were generated expressing single point mutations that abrogate the ability of Nef to interact with cellular kinases or mediate CD4 down-regulation. Infection of primary monocyte-derived macrophages (MDM) with these mutant viruses revealed that residues in the PXXP motif contribute to efficient replication. Interestingly, viruses expressing alleles of Nef defective in CD4 down-modulation activity retain wild-type levels of infectivity in single-round assays but exhibited delayed replication kinetics and grew to lower titres compared to the wild-type virus in MDM. These data suggest that efficient HIV-1 replication is dependent on the ability of Nef to interact with cellular kinases and remove CD4 from the surface of infected macrophages.
A new member of the interleukin 10-related cytokine family encoded by a poxvirus
Authors: Bartlett Nathan W. , Dumoutier Laure , Renauld Jean-Christophe , Kotenko Sergei V. , McVey Colin E. , Lee Han-Joo , Smith Geoffrey L.
Published: May 2004

Poxviruses express numerous proteins involved in manipulating the host immune response. Analysis of the primary sequence and predicted structure of the 134R protein ofYaba-like disease virus(Y134R) indicated that it is similar to cellular proteins of the IL-10 family, specifically IL-19, IL-20 and IL-24. A flag-tagged Y134R was expressed from mammalian cells and identified as a secreted, monomeric glycoprotein that stimulated signal transduction from class II cytokine receptors IL-20Rα/IL-20Rβ(IL-20R type1) and IL-22R/IL-20Rβ(IL-20R type 2). Y134R induced phosphorylation of signal transducers and activators of transcription, their translocation to the nucleus and the induction of reporter gene expression. In contrast, Y134R was unable to induce similar responses from either the IL-22 or IFN-λ(IL-28A, IL-28B, IL-29) class II cytokine receptors. To examine the role Y134R plays during a poxvirus infection, a vaccinia virus recombinant expressing Y134R was constructed and tested in a murine intranasal infection model. Compared with control viruses, the virus expressing Y134R had a reduced virulence, manifested by reduced weight loss, signs of illness and virus titres in infected organs. These results demonstrate that Y134R is a new viral member of the IL-10-related cytokine family and that its activityin vivoaffects virus virulence.
Chlamydia pneumoniae and newly diagnosed asthma: a case‐control study in 1 to 6‐year‐old children
Authors: KORPPI Matti , PALDANIUS Mika , HYVÄRINEN Anne , NEVALAINEN Aino , HUSMAN Tuula
Published: May 2004

Objective:  The aim of the study was to evaluate the association between antibodies to Chlamydia pneumoniae and the onset of asthma in children.

Methodology:  In 1996–2000, 122 children aged 1–6 years, who were treated for new asthma as inpatients or outpatients in our hospital, were recruited. For each patient, two controls, matched by age, sex and municipality, were randomly selected from the same population. In 2000, 104 serum samples were available from patients (85%) and 120 from controls (49%) for microimmunofluorescence (MIF) assay for C. pneumoniae and C. trachomatis antibodies, and for enzyme immunoassay (EIA) for C. pneumoniae antibodies.

Results:  In EIA, the median IgG concentrations were 20 EIU (EIA units) in the patients, and 16 EIU in the controls. IgG was positive (> 30 EIU) in 37 (36%) patients and in 36 (31%) controls. IgA was positive (> 12 EIU) in four (4%) patients and in eight (7%) controls. In MIF, four (4%) patients and seven (6%) controls were IgG positive, and seven were also IgA positive. IgM antibodies were detected in four children by EIA, and in none by MIF.

Conclusion:  IgG antibodies to C. pneumoniae, though common in 1 to 6‐year‐old children as detected by EIA, did not differ between newly diagnosed asthma patients and controls in this case‐control study.
Polymorphism of the promoter region of prostacyclin synthase gene in chronic thromboembolic pulmonary hypertension
Authors: AMANO Shinya , TATSUMI Koichiro , TANABE Nobuhiro , KASAHARA Yasunori , KUROSU Katsushi , TAKIGUCHI Yuichi , KASUYA Yoshitoshi , KIMURA Sadao , KURIYAMA Takayuki
Published: May 2004

Objective:  Decreased expression of prostacyclin synthase (PGIS) is observed in the lung vasculature of patients with pulmonary arterial hypertension and the biosynthesis of prostacyclin (PGI2) may be impaired in chronic thromboembolic pulmonary hypertension (CTEPH). Whether it is genetically determined or develops as the disease progresses is unclear. A variable‐number tandem repeat (VNTR) polymorphism has been detected in the 5′‐upstream promoter region of the PGIS gene. It has been demonstrated that the alleles vary in size from three to seven repeats of nine base pairs, and transcriptional activity increased with the number of repeats. The purpose of the present study was to elucidate the association between the VNTR polymorphisms of the PGIS gene and CTEPH in Japanese subjects.

Methodology:  Ninety patients with CTEPH and 144 control subjects were investigated for the presence of VNTR polymorphisms. Sixty‐two blood samples were obtained from CTEPH patients and the plasma concentrations of prostacyclin and thromboxane A2 metabolites were measured.

Results:  VNTR polymorphisms in the prostacyclin synthase gene were grouped into L alleles (five, six and seven repeats) and S alleles (three and four repeats). The overall distribution of the alleles and genotypes were not significantly different between CTEPH patients and the control subjects. The patients with the LL genotype had higher plasma concentrations of 6‐keto‐prostaglandin F1α than patients with the LS and SS genotypes.

Conclusions:  Our results suggested that the specific VNTR polymorphism in the 5′‐upstream promoter region of the PGIS gene regulated prostacyclin production, but did not seem to be associated with the development of CTEPH in this patient population.
Mycobacterium smegmatis pneumonia
Authors: ERGAN Begüm , ÇÖPLÜ Lütfi , ALP Alpaslan , ARTVINLI Mustafa
Published: May 2004

Abstract:  Mycobacterium smegmatis is a non‐tuberculous mycobacterium that is usually associated with soft tissue or wound infections in humans. Pulmonary infections secondary to this pathogen are rarely seen and occur only in patients with an underlying condition, such as lipoid pneumonia. This report presents the first case of M. smegmatis pneumonia in an otherwise healthy individual who had no predisposing condition.
Buccal mucosal urethroplasty: is it the new gold standard?
Authors: Bhargava S. , Chapple C.R.
Published: May 2004

Whilst techniques for urethral reconstruction have developed in the past few decades the quest for an ideal substitute continues. We critically review the literature on buccal mucosal grafts for substitution urethroplasty, to determine the efficacy and complications arising from its use. Buccal mucosal grafts have proved to be a versatile substitute for strictures attributable to a wide range of causes. Placing the graft dorsally appears to be more successful than ventrally and was successful in 96% of cases; after treating complex urethral strictures with two‐stage procedures about a quarter of patients required a revision after the first stage with fewer complications then when skin was used as a substitute. Thus, buccal mucosa is most likely to become the new gold standard for substitution urethroplasty and longer term results with its use are eagerly awaited.
Dorsal buccal mucosal graft urethroplasty by a ventral sagittal urethrotomy and minimal‐access perineal approach for anterior urethral stricture
Authors: Gupta N.P. , Ansari M.S. , Dogra P.N. , Tandon S.
Published: May 2004

In the mini‐review section of this issue, the use of buccal mucosal grafts in urethroplasty is described and the authors raise the point that it may now be the ‘reference’ standard. In this section, authors from New Dehli describe these technique of anterior urethral reconstruction for long strictures using buccal mucosal grafts. Their rather novel approach is of interest, and they report good results with up to 16 months of follow‐up.

OBJECTIVE

To present the technique of dorsal buccal mucosal graft urethroplasty through a ventral sagittal urethrotomy and minimal access perineal approach for anterior urethral stricture.

PATIENTS AND METHODS

From July 2001 to December 2002, 12 patients with a long anterior urethral stricture had the anterior urethra reconstructed, using a one‐stage urethroplasty with a dorsal onlay buccal mucosal graft through a ventral sagittal urethrotomy. The urethra was approached via a small perineal incision irrespective of the site and length of the stricture. The penis was everted through the perineal wound. No urethral dissection was used on laterally or dorsally, so as not to jeopardize the blood supply.

RESULTS

The mean (range) length of the stricture was 5 (3–16) cm and the follow‐up 12 (10–16) months. The results were good in 11 of the 12 patients. One patient developed a stricture at the proximal anastomotic site and required optical internal urethrotomy.

CONCLUSION

Dorsal buccal mucosal graft urethroplasty via a minimal access perineal approach is a simple technique with a good surgical outcome; it does not require urethral dissection and mobilization and hence preserves the blood supply.
Nocturia and depression
Authors: Asplund R. , Henriksson S. , Johansson S. , Isacsson G.
Published: May 2004

OBJECTIVE

To assess the possible relationship between major depression (MD) and nocturia.

SUBJECTS AND METHODS

An unselected group of adult men and women, living in the city of Östersund, Sweden, were sent a postal questionnaire containing questions on somatic and mental health, sleep, sleepiness and nocturia. For depression diagnostics the Major Depression Inventory (MDI) was used.

RESULTS

The mean (sd) ages of the men and women were 48.0 (18.2) and 50.1 (19.1) years, respectively. Two or more nocturnal micturition episodes occurred in 15.6% of the men and 16.5% of the women. In a multiple logistic regression analysis independent correlates (with 95% confidence intervals) for ≥ 2 nocturnal voids in men were: MD 6.5 (2.6–15.6); health, fair vs good, 1.8 (1.0–3.9); health, poor vs good, 2.7 (1.1–4.8); age, 60–74 vs 18–29, 3.6 (1.5–8.4) and age ≥ 75 vs 18–29 6.7 (2.6–17.4); and in women: MD 2.8 (1.3–6.3); health, fair vs good, 1.9 (1.1–3.2); health, poor vs good, 4.3 (2.6–8.2); age, 60–74 vs 18–29, 3.8 (1.8–7.8), and age ≥ 75 vs 18–29, 8.6 (4.0–18.6). Age < 60 years was deleted by the logistic model in both sexes.

CONCLUSION

MD is associated with a six‐fold increase in nocturia in men and a three‐fold increase in women, after accounting for age and health. Possible pathogenetic mechanisms of this relationship may involve both increased nocturnal diuresis via a disturbed 24‐h rhythm of antidiuretic hormone secretion, and a decrease in nocturnal bladder capacity through a central and/or peripheral serotonergic effect.
Concurrent radical retropubic prostatectomy and inguinal hernia repair through a modified Pfannenstiel incision
Authors: Manoharan M. , Gomez P. , Soloway M.S.
Published: May 2004

Authors from Miami describe their technique of concurrent radical retropubic prostatectomy and inguinal hernia repair through a modified Pfannenstiel incision. They found this approach to be ideal for performing both operations at the same time, allowing a tension‐free mesh hernia repair with excellent exposure of the pelvic structures.

An interesting paper from authors in New York reviews patients who had both bladder and lung cancer. Their findings are important in terms of a potential guideline to urologists who have patients with these conditions, and are, for example, faced with the decision as to whether they should operate on someone who has a history of lung cancer but now has bladder cancer.

In another paper from the New York area, authors describe the effect on morbidity and mortality of bone metastasis in patients with RCC. They found the effect to be considerable, and suggest a possible role for bisphosphonates.

OBJECTIVE

To describe a technique for concurrent radical retropubic prostatectomy (RRP) and inguinal hernioplasty, using a modified Pfannenstiel incision.

PATIENTS AND METHODS

RRP is usually done through a midline lower abdominal incision but some patients with localized prostate cancer have an inguinal hernia. Concurrent inguinal hernia repair at the time of RRP with the usual method is only possible by either a preperitoneal mesh repair or formal hernioplasty, requiring an additional incision(s). A 10–12 cm Pfannenstiel incision is made along the pubic hairline centred over the pubic symphysis, and a ‘Y’‐shaped incision in the rectus sheath. The rectus muscle is split vertically along the midline, followed by RRP. After removing the prostate and completing the anastomosis, the surgeon identifies the inguinal canal along the inferior and lateral aspect of the transverse incision and uses a formal tension‐free hernioplasty with a 3 × 5 cm polypropylene mesh. We used this technique in fifteen concurrent inguinal hernioplasties (two bilateral hernias and thirteen unilateral) at the time of RRP, with no additional incisions, using the formal tension‐free Lichtenstein technique. One patient with bilateral hernias had a right indirect inguinal hernia, and all the remaining men had a direct inguinal hernia.

RESULTS

All patients were discharged 2 days after surgery, with no complications associated with the procedure and no recurrences; however, the follow‐up was short (mean 5.5 months).

CONCLUSION

A modified Pfannenstiel incision is ideal for concurrent RRP and inguinal hernioplasty, providing excellent exposure of the pelvic structures and allowing the surgeon to use a formal tension‐free mesh hernioplasty through the same incision. Wound healing and cosmetic results are excellent.
Risk factors for renal cysts
Authors: Terada N. , Arai Y. , Kinukawa N. , Yoshimura K. , Terai A.
Published: May 2004

OBJECTIVE

To examine the risk factors for renal cysts in a large population‐based health survey, as we previously reported that the prevalence of renal cyst increases with age, there is a difference in incidence between the sexes and other studies show an association between renal cysts and hypertension.

SUBJECTS AND METHODS

Data were collected on 17 914 individuals who participated in a multiphase health‐screening programme at our institution in 2000. Ultrasonography was used for diagnosing renal cysts. Logistic analysis was used to examine various risk factors for renal cyst, including sex, age, serum creatinine, hypertension, hypercholesterolaemia, diabetes mellitus, and smoking habits. Hypertension was defined as a systolic blood pressure of > 140 mmHg, a diastolic blood pressure of > 90 mmHg, or current use of antihypertensive medication. In 45 patients with renal cysts who were followed for a mean (range) of 6 (4–7) years the sequential changes in the size of the cysts and the systolic blood pressure were plotted in relation to age. The relationship of the mean changes in these variables was also examined.

RESULTS

The prevalence of renal cysts was 9.9%, ranging from 3.8% for subjects in their third decade to 18.5% in their sixth. Cysts were detected in 13.0% of men and 5.8% of women (P < 0.001). The mean serum creatinine was 83 mg/L in those with cysts and 76 mg/L in those without (P < 0.001); the respective mean systolic blood pressure was 123 and 118 mmHg (P < 0.001). Multivariate logistic regression analysis showed that age (P < 0.001), sex (P < 0.001), hypertension (P = 0.0022) and serum creatinine (P = 0.021) had a significant influence on the occurrence of renal cysts. Enlargement of the cysts was not correlated with the increase in blood pressure.

CONCLUSIONS

The risk factors for a renal cyst are age, male gender, renal dysfunction and hypertension. Hypertension might cause renal dysfunction, which leads to the development of renal cysts.
Identification of multidrug resistance‐associated protein 1 and glutathione as multidrug resistance mechanisms in human prostate cancer cells: chemosensitization with leukotriene D4 antagonists and buthionine sulfoximine
Authors: Van Brussel J.P. , Oomen M.A. , Vossebeld P.J.M. , Wiemer E.A.C. , Sonneveld P. , Mickisch G.H.J.
Published: May 2004

OBJECTIVE

To assess the involvement of the multidrug resistance‐associated protein 1 (MRP1) and the glutathione pathway in the multidrug resistant (MDR) phenotype of prostate cancer in vitro.

MATERIALS AND METHODS

Chemoselection of human prostate cancer cell lines PC3 and DU145 with etoposide resulted in the resistant cell lines PC3‐R and DU‐R. Resistance against etoposide, doxorubicin and vincristine, and its reversal with leukotriene D4 antagonists MK‐571 and zafirlukast, and buthionine sulfoximine (BSO), was assessed using tetrazolium‐dye viability assays. Western blot analysis of MRP1 expression and glutathione content were measured, and MRP1 function assessed in fluorescence assays.

RESULTS

MRP1 was increased in the MDR models; the glutathione content was significantly higher in PC3‐R but there was no increase in glutathione in DU‐R. Adding non‐toxic doses of MK‐571, zafirlukast or BSO significantly increased the sensitivity of the MDR models to cytotoxic drugs. MRP1 function was inhibited with MK‐571 in the MDR models.

CONCLUSION

MRP1 and glutathione mediate MDR in newly developed prostate cancer models.
Papillary urothelial neoplasm of low malignant potential: reliability of diagnosis and outcome
Authors: Campbell P.A. , Conrad R.J. , Campbell C.M. , Nicol D.L. , MacTaggart P.
Published: May 2004

OBJECTIVE

To determine the ability of pathologists to reproducibly diagnose a newly defined lesion, i.e. the papillary urothelial neoplasm of low malignant potential (PUNLMP) using the published criteria, defined by the 1998 World Health Organisation/International Society of Urological Pathology (WHO/ISUP) classification system; in addition, debate remains about the clinical behaviour of these lesions, thus the rates of recurrence and progression of PUNLMP lesions were assessed and compared with low‐grade papillary urothelial carcinomas (LG‐PUC) and high‐grade (HG‐PUC) over a 10‐year follow‐up.

PATIENTS AND METHODS

Forty‐nine cases of superficial bladder cancer (G1–3 pTa) representing an initial diagnosis of transitional cell carcinoma made in 1990 were identified and re‐graded using the 1998 WHO/ISUP classification by two pathologists. Inter‐observer agreement was assessed using Cohen weighted κ statistics. After re‐classification the clinical follow‐up was reviewed retrospectively, and episodes of recurrence and progression recorded.

RESULTS

The inter‐observer agreement was moderate, regardless of whether one (κ 0.45) or two (κ 0.60) pathologists were used to grade these lesions. Re‐classification identified 12 PUNLMP, 28 LG‐PUC and nine HG‐PUC. PUNLMP lesions recurred in 25% (3/12) of cases; no progression was documented. Recurrence rates were 75% (21/28) and 67% (6/9) for LG‐ and HG‐PUC, respectively, and progression rates were 4% (1/28) and 22% (2/9).

CONCLUSION

The 1998 WHO/ISUP classification of urothelial neoplasms can be reproducibly applied by pathologists, with a moderate level of agreement. There is evidence that PUNLMP lesions have a more indolent clinical behaviour than urothelial carcinomas. However, the risk of recurrence and progression remains, and clinical monitoring of these patients is important.
Prevention of recurrent bacterial cystitis by intravesical administration of hyaluronic acid: a pilot study
Authors: Constantinides C. , Manousakas T. , Nikolopoulos P. , Stanitsas A. , Haritopoulos K. , Giannopoulos A.
Published: May 2004

OBJECTIVES

To assess the effect of bladder instillations of hyaluronic acid (HA) on the rate of recurrence of urinary tract infection (UTI).

PATIENTS AND METHODS

Forty women (mean age 35 years) with a history of recurrent UTI received intravesical instillations of HA (40 mg in 50 mL phosphate‐buffered saline) once weekly for 4 weeks then once monthly for 4 months. The UTI status was assessed over a prospective follow‐up of 12.4 months and compared with the rates of UTI before instillation, determined by a retrospective review of patient charts covering 15.8 months.

RESULTS

After HA treatment no patients had a UTI during the 5‐month treatment phase and 28 (70%) were recurrence‐free at the end of the follow‐up. The mean (sd) rate of UTI per patient‐year was 4.3 (1.55) before treatment and 0.3 (0.55) afterward (P < 0.001). The median time to recurrence after HA treatment was 498 days, compared with 96 days beforehand (P < 0.001). The tolerability was excellent, as side‐effects were limited to nine patients who reported mild bladder irritation; no patient interrupted the treatment.

CONCLUSIONS

In this preliminary study, bladder instillations of HA had a significant effect on the rate of UTI in women with a history of recurrent UTIs. The bladder instillation of HA is an acceptable and promising therapeutic alternative in patients with recurrent UTI. Expanded placebo controlled clinical trials examining this application of HA are currently underway.
Transurethral incision of duplex system ureteroceles in neonates: does it increase the need for secondary surgery in intravesical and ectopic cases?
Authors: Castagnetti M. , Cimador M. , Sergio M. , De Grazia E.
Published: May 2004

OBJECTIVE

To evaluate the relevance of ureterocele ectopia and associated reflux on the outcome of duplex system ureteroceles (DSU) after neonatal transurethral incision (TUI).

PATIENTS AND METHODS

The study included 41 neonates with a diagnosis of DSU; the ureterocele was ectopic in 24 (58%). Before TUI, vesico‐ureteric reflux (VUR) was present in 13 lower moieties (32%) and seven contralateral ureters (17%). TUI was always performed within the first month of life. The follow‐up and management were tailored for each patient from the findings at ultrasonography, voiding cysto‐urethrography and renal scintigraphy. Results of intravesical and ectopic DSU were compared using Fisher's exact test.

RESULTS

TUI was effective in allowing ureteric decompression in all but one patient (2.4%). After TUI, VUR ceased in six lower ipsilateral moieties and in two contralateral ureters, while new VUR occurred in three contralateral kidneys. De novo VUR in the punctured moiety appeared in 13 cases (32%). Nine upper poles were not functioning. Twenty‐one patients (51%) required secondary surgery. Ureteric reimplantation was indicated exclusively for reflux in the punctured moiety in only in two cases (5%), while in a further two iatrogenic reflux in a nonfunctioning upper moiety required total heminephro‐ureterectomy. There was no significant difference between intravesical and ectopic ureteroceles in the occurrence of VUR in the punctured moiety, rate of nonfunctioning upper poles or need for secondary surgery.

CONCLUSIONS

About half of the patients with a DSU need secondary surgery, but this is rarely indicated for de novo reflux in the punctured moiety only. The need for secondary surgery was greater whenever there was associated reflux before endoscopic incision. There was no difference in the outcome of intravesical and ectopic ureteroceles and such distinction seems no longer to be of clinical relevance.
Predictors of androgen independence in metastatic prostate cancer
Authors: Sim H.G. , Lau W.K.O. , Cheng C.W.S.
Published: May 2004

OBJECTIVE

To assess the factors that influence the onset of androgen independence (AI, which heralds a dismal outcome) in patients with metastatic prostate carcinoma.

PATIENTS AND METHODS

The records of 361 consecutive patients with prostate carcinoma diagnosed and treated in the authors’ institution from 1 January 1996 to 31 December 1999 were reviewed retrospectively; 92 with metastatic prostate carcinoma were assessed (median age 71.0 years, range 42–93). Patients were included if they developed metastatic disease from prostate cancer at the time of diagnosis. The nadir for prostate specific antigen (PSA) level was defined as the date of the lowest PSA level after hormonal therapy, and AI was defined as the date of the third consecutive PSA increase above the nadir value by any threshold.

RESULTS

The median Gleason sum was 8 and the modal Gleason score 4 + 5. The median (range) pretreatment PSA level was 274.0 (1.3–2179) ng/mL. Of the 92 men, 57 (62%) attained a nadir PSA, including 23 with a nadir of < 2 ng/mL; 32 (35%) progressed to AI within 2 years and 27% reached a nadir PSA but did not develop AI. The mean (sd) time from diagnosis to the nadir PSA was 13.7 (11.8) months, while the mean time from diagnosis to progression to AI was 30.3 (15.6) months. Univariate analysis showed that a nadir PSA level after treatment of ≥ 1 ng/mL (P = 0.0128) was an early predictor of progression to AI; a nadir PSA level of ≥ 2 ng/mL (P = 0.0216) was a predictor of poor overall survival.

CONCLUSION

Failure to attain a nadir PSA of < 1 ng/mL after treatment predicts progression to AI and a nadir PSA of > 2 ng/mL predicts poorer overall survival. The development of skeletal events predicts the onset of AI but occurs late in the disease and is unsuitable as an early prognostic marker.
Linear Stability of Parallel Inviscid Flows of Shallow Water and Bubbly Fluid
Authors: Gavrilyuk S. L. , Teshukov V. M.
Published: May 2004

The extensions of Howard's semicircle theorem, Squire's transformation, and Rayleigh's stability criterion are obtained for the model of dispersive shallow water waves and the model of bubbly fluid.
Discrete Curves in and the Toda Lattice
Authors: Hoffmann Tim , Kutz Nadja
Published: May 2004

In this paper, we investigate flows on discrete curves in , and . A novel interpretation of the one‐dimensional Toda lattice hierarchy and reductions thereof as flows on discrete curves are given.
Elastic Behavior of Cross-Linked and Bundled Actin Networks
Authors: Gardel M. L. , Shin J. H. , MacKintosh F. C. , Mahadevan L. , Matsudaira P. , Weitz D. A.
Published: May 2004

Networks of cross-linked and bundled actin filaments are ubiquitous in the cellular cytoskeleton, but their elasticity remains poorly understood. We show that these networks exhibit exceptional elastic behavior that reflects the mechanical properties of individual filaments. There are two distinct regimes of elasticity, one reflectingbendingof single filaments and a second reflectingstretchingof entropic fluctuations of filament length. The mechanical stiffness can vary by several decades with small changes in cross-link concentration, and can increase markedly upon application of external stress. We parameterize the full range of behavior in a state diagram and elucidate its origin with a robust model.
A Family with Severe Insulin Resistance and Diabetes Due to a Mutation in
AKT2
Authors: George Stella , Rochford Justin J. , Wolfrum Christian , Gray Sarah L. , Schinner Sven , Wilson Jenny C. , Soos Maria A. , Murgatroyd Peter R. , Williams Rachel M. , Acerini Carlo L. , Dunger David B. , Barford David , Umpleby A. Margot , Wareham Nicholas J. , Davies Huw Alban , Schafer Alan J. , Stoffel Markus , O'Rahilly Stephen , Barroso Inês
Published: May 2004

Inherited defects in signaling pathways downstream of the insulin receptor have long been suggested to contribute to human type 2 diabetes mellitus. Here we describe a mutation in the gene encoding the protein kinase AKT2/PKBβ in a family that shows autosomal dominant inheritance of severe insulin resistance and diabetes mellitus. Expression of the mutant kinase in cultured cells disrupted insulin signaling to metabolic end points and inhibited the function of coexpressed, wild-type AKT. These findings demonstrate the central importance of AKT signaling to insulin sensitivity in humans.
A New Synthesis or Just The New Synthesis?
Authors: Peterson A. Townsend
Published: May 2004

The Speciation and Biogeography of Birds
. by Ian Newton. Academic Press (Elsevier), San Diego, 2003. 680 pp. $75, £49.95. ISBN 0-12-517375-X.

Newton offers a comprehensive overview of the origins and distribution of bird species and discusses the factors that shape these patterns.
Sentinel lymph node investigation in melanoma: detailed analysis of the yield from step sectioning and immunohistochemistry
Authors: Gietema H A , Vuylsteke R J C L M , de Jonge I A , van Leeuwen P A M , Molenkamp B G , van der Sijp J R M , Meijer S , van Diest P J
Published: May 2004

Aims:To evaluate in detail the extent to which step sectioning and immunohistochemical examination of sentinel lymph nodes (SLNs) in patients with melanoma reveal additional node positive patients, to arrive at a sensitive yet workable protocol for histopathological SLN examination.

Methods:The study comprised 29 patients with one or more positive SLN after a successful SLN procedure for clinical stage I/II melanoma. SLNs were lamellated into pieces of approximately 0.5 cm in size. One initial haematoxylin and eosin (H&E) stained central cross section was made for each block. When negative, four step ribbons were cut at intervals of 250 μm. One section from each ribbon was stained with H&E, and one was used for immunohistochemistry (IHC).

Results:When taking the cumulative total of detected metastases at level 5 as 100%, the percentage of SLN positive patients increased from 79%, 83%, 83%, 90% to 93% in the H&E sections through levels 1–5, and with IHC these values were 83%, 86%, 90%, 97%, and 100%, respectively. One of six patients in whom metastases were detected at levels 2–5 only had metastases in the subsequent additional lymph node dissection.

Conclusions:Multiple level sectioning of SLNs (five levels at 250 μm intervals) and the use of IHC detects additional metastases up to the last level in melanoma SLNs. Although more levels of sectioning might increase the yield even further, this protocol ensures a reasonable workload for the pathologist with an acceptable sensitivity when compared with the published literature.
Resistance to scrapie in PrP ARR/ARQ heterozygous sheep is not caused by preferential allelic use
Authors: Caplazi P A , O’Rourke K I , Baszler T V
Published: May 2004

Background:In sheep, susceptibility to scrapie, which is similar to human prion diseases such as Kuru and variant Creutzfeldt-Jakob disease (vCJD), is determined by prion protein (PrP) gene (Prnp) polymorphisms. Sheep with genotype ARQ/ARQ, denoting polymorphisms at codons 136, 154, and 171, are susceptible, whereas those with genotypes ARR/ARQ and ARR/ARR are resistant, indicating dominance of ARR over the ARQ allele.

Aims:Based on familial CJD E200K, 129V, where preferential use of the 200E allele in EK heterozygous individuals confers resistance, heterozygous ARR/ARQ sheep were used to test the hypothesis that resistance is caused by preferential use of the ARR allele.

Methods:After assessment of equivalent PrP expression across genotypes, allele use was analysed by sequencing reverse transcription polymerase chain reaction derived DNA clones containing the Prnp gene coding sequence.

Results:The ARR to ARQ ratio was 1.1 in 133 clones, representing Prnp mRNA from three ARR/ARQ sheep, indicating equal use of both alleles.

Conclusions:Dominance of the resistant associated allele in sheep scrapie involves mechanisms other than the absence of PrP derived from the disease associated ARQ allele.
High resolution microarray comparative genomic hybridisation analysis using spotted oligonucleotides
Authors: Carvalho B , Ouwerkerk E , Meijer G A , Ylstra B
Published: May 2004

Background:Currently, comparative genomic hybridisation array (array CGH) is the method of choice for studying genome wide DNA copy number changes. To date, either amplified representations of bacterial artificial chromosomes (BACs)/phage artificial chromosomes (PACs) or cDNAs have been spotted as probes. The production of BAC/PAC and cDNA arrays is time consuming and expensive.

Aim:To evaluate the use of spotted 60 mer oligonucleotides (oligos) for array CGH.

Methods:The hybridisation of tumour cell lines with known chromosomal aberrations on to either BAC or oligoarrrays that are mapped to the human genome.

Results:Oligo CGH was able to detect amplifications with high accuracy and greater spatial resolution than other currently used array CGH platforms. In addition, single copy number changes could be detected with a resolution comparable to conventional CGH.

Conclusions:Oligos are easy to handle and flexible, because they can be designed for any part of the genome without the need for laborious amplification procedures. The full genome array, containing around 30 000 oligos of all genes in the human genome, will represent a big step forward in the analysis of chromosomal copy number changes. Finally, oligoarray CGH can easily be used for any organism with a fully sequenced genome.
Tissue invasiveness and non-acidic pH in human candidiasis correlate with “in vivo” expression byCandida albicansof the carbohydrate epitope recognised by new monoclonal antibody 1H4
Authors: Monteagudo C , Viudes A , Lazzell A , Martinez J P , Lopez-Ribot J L
Published: May 2004

Background:The morphogenetic conversion between yeast and hyphal growth forms appears to be crucial in the pathogenesis of invasive candidiasis, and can be regulated by environmental signals such as extracellular pH.

Aims:To characterise the epitope recognised by monoclonal antibody 1H4, and to evaluate the expression of its corresponding epitope inCandida albicanscells under different conditions of pH and temperature, and “in vivo”, in tissue samples from patients with human candidiasis.

Methods:Monoclonal antibody 1H4 was generated against the 58 kDa cell wall mannoprotein ofC albicans(mp58), and was further characterised by immunoblot analysis, periodate treatment of the antigenic preparations, and agglutination experiments ofC albicansstrains 3153A, SC5314, and 412, cultured under different environmental conditions (growth media and pH). An immunohistochemical study was performed in 24 human tissue samples from patients with mucocutaneous and systemic candidiasis.

Results:1H4 recognises a pH sensitive carbohydrate epitope on the surface ofC albicanscells, and this epitope is not restricted to mp58, but is shared with other cell wall mannoproteins. Immunohistochemical findings indicated that expression of the 1H4 epitope onC albicanscells in tissue sections from human candidiasis correlates with tissue invasion and pH of the niche. 1H4 immunoreactivity was also found in candida remnants within macrophages.

Conclusions:The fact that 1H4 epitope expression selectively identifies invasive forms ofC albicans, in addition to candida remnants within macrophages, supports its potential value in the diagnosis and management of human candidiasis.
DoToxoplasma gondiiRH strain tachyzoites evolve during continuous passage?
Authors: Mavin S , Joss A W L , Ball J , Ho-Yen D O
Published: May 2004

Aim:To examine three lineages ofToxoplasma gondiiRH strain in terms of performance in the dye test, culture, and gene expression.

Methods:Historical data (culture growth and performance in the dye test) from three lineages of RH strain tachyzoites (B, J, and Q) that had been continuously cultured in HeLa cells was assessed. Tachyzoite harvests obtained during continuous cell culture were retrieved from liquid nitrogen and cultured in HeLa cells, providing mRNA that was extracted and used to study gene expression using random amplified polymorphic DNA analysis at different stages of lineage adaptation to continuous culture.

Results:The B and Q lineages consistently produced tachyzoites that were successfully used in the dye test and their gene expression was stable after multiple passages. The J lineage had unpredictable growth, tachyzoites unsuitable for use in the dye test, and changing gene expression with multiple passage.

Conclusion:This study has explained some anomalies in the performance of different stocks ofT gondii, and suggests that lineages that are still evolving in cell culture should be avoided.
Use of multiple displacement amplification to amplify genomic DNA before sequencing of the α and β haemoglobin genes
Authors: Mai M , Hoyer J D , McClure R F
Published: May 2004

Aims:To evaluate the technique of multiple displacement amplification (MDA) for whole genome amplification from small volume blood samples before sequencing in a clinical test to identify haemoglobin gene mutations.

Methods:Phage φ29 DNA polymerase was used to perform MDA, starting with either 1 μl of blood or 1 ng of previously isolated blood DNA from 23 patients. The amplified products were then evaluated using a clinical test that involves sequencing the haemoglobin genes to detect mutations. The results were compared with the current clinical test method that uses genomic DNA isolated using column based technology.

Results:The MDA technique produced large quantities (theoretically approximately 2 mg) of DNA. The amplification procedure was extremely easy and took about four hours (less than one hour of hands on technician time and three hours for amplification). When MDA products were used in the same clinical test protocol as genomic DNA isolated using column technology, there was 100% concordance for detection of a variety of point mutations in the α1, α2, and β globin genes.

Conclusions:The MDA technique is useful for overcoming the problem of insufficient genomic DNA in clinical specimens requiring haemoglobin gene sequencing and could be useful for other clinical applications.
Treatment difficulties of a listerial rhombencephalitis in an adult patient allergic to penicillins
Authors: Popescu G A , Saquepée M , Poisson D , Prazuck T
Published: May 2004

Rhombencephalitis is not a rare presentation of listerial central nervous system infections in healthy adults. This report describes a case with several management difficulties linked to antibiotic related adverse events, pointing to alternative solutions to aminopenicillins. In addition, the role of dexamethasone in the management of inflammation and neurological symptoms is discussed.
Systemic mastocytosis with associated clonal haematological non-mast cell lineage diseases: a histopathological challenge
Authors: Horny H-P , Sotlar K , Sperr W R , Valent P
Published: May 2004

Aims:Although systemic mastocytosis (SM) with an associated clonal haematological non-mast cell lineage disease (SM-AHNMD) is a major subtype of SM, little is known about its frequency among myelogenous neoplasms, and mastocytosis in particular, or about AHNMD subtype frequencies.

Methods:Approximately 19 500 routine bone marrow biopsies were evaluated. Immunostaining with antibodies against tryptase, KIT, and CD25 and molecular analysis for detection of C-KIT point mutations were performed in approximately 550/4100 myelogenous malignancies including mastocytosis, almost all subtypes of myelodysplastic syndrome (MDS), myelodysplastic/myeloproliferative syndrome (MDS/MPD), MPD, and acute myeloid leukaemia (AML).

Results:SM was rare—it was diagnosed in only 64 bone marrows (0.3%) and made up 1.5% of myelogenous tumours. SM-AHNMD was the second most frequent subtype (20). SM-AHNMD was never included in the clinical differential diagnoses and was confirmed histologically in most cases only after appropriate immunostaining. The abnormal mast cell phenotype was confirmed by immunohistochemical demonstration of tryptase and CD25 coexpression. The following associated haematological neoplasms were found: MDS/MPS, AML, MPS, MDS, plasma cell myeloma, and unclassifiable myelogenous malignancy. C-KIT point mutations were detected in 16 of 20 cases.

Conclusions:SM-AHNMD can be diagnosed histologically in bone marrow trephines only after immunostaining with antibodies against tryptase, KIT, and CD25. Eighteen of 20 AHNMDs were of myeloid origin. C-KIT point mutations were present in 16 of 20 cases. The prognostic relevance of detecting SM associated with another haematological neoplasm remains unclear, but mast cell resistance to most cytoreductive agents is of major importance for treatment planning.
Heterogeneity of the fibre sequence in subgenus C adenoviruses
Authors: Adhikary A K , Banik U , Numaga J , Suzuki E , Inada T , Okabe N
Published: May 2004

Aims:To determine the nucleotide sequences of adenovirus (Ad) types 1 and 6 fibre genes; to clarify the molecular basis of the distinct haemagglutination properties of subgenus C Ads and their phylogenetic relations.

Methods:Human Ad1 and Ad6 fibre genes were sequenced from genomic DNA by direct sequencing. Primer selection was based on alignment of the fibre gene of human Ad serotypes Ad2 and Ad5. Fibre based subgenus C specific polymerase chain reaction (PCR) was performed to check for deletions in field isolates of Ad6, as revealed by sequence analysis of the Ad6 prototype. A phylogenetic tree was constructed from the predicted amino acid (AA) sequences of the fibre gene of important Ads.

Results:Ad1 and Ad6 comprise 1746 and 1584 nucleotides, encoding 582 and 528 AA, respectively. Ad6 showed deletions in motifs 15–17 (51 AA) of the shaft when compared with Ad1, Ad2, and Ad5. Subgenus C specific PCR with both prototype and field isolates also showed deletions in Ad6. In the shaft and knob, AA homology was 58.82–72.91% and 68.89–74.59%, respectively. The tail was 100% conserved. Phylogenetically, Ad1 and Ad6, including Ad2 and Ad5, formed a subgenus specific cluster, like other serotypes.

Conclusions:The fibre gene (including the knob region) of subgenus C Ads is heterogeneous, providing the molecular basis for lack of crossreactivity in the haemagglutination inhibition test. This heterogeneity could be helpful in fibre based genotyping of subgenus C field isolates. Phylogeny might be useful for subgenus specific identification of important field strains.