bluuebunny/crossref_metadata_2025 huggingface dataset:
Differential Suppression of Human Cervical Cancer Cell Growth by Adenovirus Delivery of p53 in vitro: Arrest Phase of Cell Cycle Is Dependent on Cell Line
Authors: Ahn Woong Shick , Han You Jin , Bae Su Mi , Kim Tae‐Hyung , Rho Min Seok , Lee Joon Mo , Namkoong Sung Eun , Park Yong Seok , Kim Chong Kook , Sin Jeong‐Im
Published: August 2005

It has been reported that overexpression of wild‐type p53 protein induces suppression of tumor cell growth in vivo and in vitro. In this study, we further evaluated the differential effects of p53 delivered in an adenovirus vector on the cell growth, apoptosis and cell cycle progression in cervical cancer cell lines. We constructed a recombinant adenovirus expressing p53 and then delivered this into cervical carcinoma cell lines (CaSki, SiHa, and HeLa, HeLaS3) along with adenovirus expressing β‐galactosidase as a negative control. Adenovirus‐delivered p53 overexpression resulted in a more significant suppression of cell growth in HPV 18‐infected cells (HeLa and HeLaS3) and a lesser suppression in HPV 16‐infected cells (CaSki and SiHa). However, no suppression was observed in cells infected with a negative control virus. p53 overexpression also induced apoptosis and cell cycle arrest, as determined by annexin V and propidium iodide staining. In particular, the cell cycle was arrested in the G2/M phase in CaSki cells. In contrast, cell cycles were arrested in the G1 phase in HeLa cells, suggesting that the arrest phase is dependent upon the cervical cancer cell line. Taken together, these data support the idea that overexpressed p53 protein plays a differential role in suppressing cervical cancer cell growth through apoptosis and cell cycle arrest in either G1 or G2/M phase, depending on the cancer cell line.
Thalidomide for the Treatment of Refractory Multiple Myeloma: Association of Plasma Concentrations of Thalidomide and Angiogenic Growth Factors with Clinical Outcome
Authors: Kakimoto Tsunayuki , Hattori Yutaka , Okamoto Shinichiro , Sato Norihide , Kamata Tamihiro , Yamaguchi Masaya , Morita Kunihiko , Yamada Taketo , Takayama Nobuyuki , Uchida Hideo , Shimada Naoki , Tanigawara Yusuke , Ikeda Yasuo
Published: August 2005

Recent reports showed that thalidomide has anti‐angiogenic activity and is effective for the treatment of refractory multiple myeloma (MM). We examined the relationship between the clinical efficacy and adverse effects of thalidomide and the plasma concentrations of this drug as well as angiogenic growth factors in refractory MM. Ten out of twenty‐four evaluable patients (42%) showed more than 25% reduction of M‐protein, and eight (33%) achieved more than 50% reduction. These changes were associated with restoration of anemia and recovery of normal immunoglobulin level. Somnolence and headache, constipation, peripheral neuropathy and skin rash were frequently observed, but were well tolerated. However, grade 2–4 severe neutropenia was also observed in nine cases. These adverse effects other than neutropenia occurred more frequently in the patients with higher plasma concentrations of thalidomide (≥2.0 μg/ml at 12 h after the last administration) and were readily alleviated by dose reduction. In contrast, neutropenia developed regardless of the plasma concentration. Plasma concentrations of angiogenic growth factors were frequently elevated before treatment. After thalidomide treatment, these growth factor levels tend to decrease to near‐normal ranges in responders but were still high in most non‐responders. After thalidomide treatment, plasma vascular endothelial growth factor (VEGF) level was significantly reduced in responders (P=0.025), but not in non‐responders (P=0.37). Reduction of plasma VEGF level might be an important indicator for anti‐myeloma effect of thalidomide.
Dose‐response Trial of Lactoferrin in Patients with Chronic Hepatitis C
Authors: Okada Shuichi , Tanaka Katsuaki , Sato Tosiya , Ueno Hideki , Saito Satoru , Okusaka Takuji , Sato Keiko , Yamamoto Seiichiro , Kakizoe Tadao
Published: August 2005

Hepatitis C virus (HCV) is one of the most common causes of chronic hepatitis. Interferon is presently the only effective treatment for chronic hepatitis C (CH‐C), though its effectiveness is limited. Lactoferrin (LF), which is an 80‐kDa, iron‐binding glycoprotein, has several biological activities including anti‐viral activity, and it was recently reported to inhibit HCV infection in cultured human hepatocytes. The present trial was designed to assess the relationship between the dose of bovine LF (bLF) and the effect of bLF on serum alanine aminotransaminase (ALT) and HCV RNA levels in patients with CH‐C. Forty‐five patients entered at each of the three dose levels (bLF of 1.8, 3.6, and 7.2 g/day) received orally an 8‐week course of bLF. There was no significant relation between the dose of bLF and the effect of bLF on serum ALT or HCV RNA levels. Biochemical (a 50% or greater decrease in the serum ALT level) and virological (a 50% or greater decrease in HCV RNA level) responses were observed in two and four patients, respectively, but all responders relapsed during the follow‐up period after bLF treatment. The bLF treatment was generally well tolerated, and no patient had any serious adverse event. In conclusion, the excellent tolerance and potential anti‐HCV activity of bLF shown in this trial suggest that further trials using a large number of patients are mandatory. We are currently conducting a double‐blind randomized controlled trial comparing bLF with placebo to clarify the anti‐HCV activity of bLF in patients with CH‐C.
DNA methylation of genes linked to retinoid signaling in squamous cell carcinoma of the esophagus: DNA methylation of CRBP1 and TIG1 is associated with tumor stage
Authors: Mizuiri Hirozumi , Yoshida Kazuhiro , Toge Tetsuya , Oue Naohide , Aung Phyu Phyu , Noguchi Tsuyoshi , Yasui Wataru
Published: August 2005

Hypermethylation of CpG islands is associated with the silencing of various tumor suppressor genes. Retinoic acid receptor‐beta (RAR‐beta), cellular retinol‐binding protein 1 (CRBP1), and tazarotene‐induced gene 1 (TIG1) have been linked to retinoic acid signaling. Little is known about the involvement of these three genes in esophageal squamous cell carcinoma (ESCC). In this study, we investigated the methylation status of these genes and analyzed the role of methylation of their DNA in ESCC. Methylation‐specific polymerase chain reaction (PCR) was performed to study the methylation of CpG islands in 28 ESCC (stages I, II, and III) and 10 samples of corresponding non‐neoplastic mucosa. The mRNA expression levels of the three genes were measured by quantitative reverse transcription‐PCR. DNA hypermethylation of RAR‐beta was found in seven (25.0%) of the 28 ESCC, of CRBP1 in five (17.9%), and of TIG1 in five (17.9%). DNA methylation of RAR‐beta was identified in one of 10 samples of corresponding non‐neoplastic mucosa (10.0%), whereas no DNA methylation of CRBP1 or TIG1 was detected. In total, at least one of the three genes was hypermethylated in 12 (42.9%) ESCC. Reduced expression of RAR‐beta, CRBP1, and TIG1 was found in 14 (50.0%), 15 (53.6%), and 13 (46.4%) ESCC, respectively. DNA methylation of each gene was significantly associated with reduced expression of the respective mRNA. No correlation was found between the DNA methylation status of RAR‐beta and clinicopathological factors such as depth of invasion, lymph node metastasis, or tumor stage. In contrast, DNA methylation of both CRBP1 and TIG1 was observed only in stage III ESCC. These results show that inactivation of the retinoic acid signaling‐associated genes RAR‐beta, CRBP1, and TIG1 by DNA methylation occurs frequently in ESCC. (Cancer Sci 2005; 96: 571–577)
Dietary intakes of fat and fatty acids and risk of breast cancer: A prospective study in Japan
Authors: Wakai Kenji , Tamakoshi Koji , Date Chigusa , Fukui Mitsuru , Suzuki Sadao , Lin Yingsong , Niwa Yoshimitsu , Nishio Kazuko , Yatsuya Hiroshi , Kondo Takaaki , Tokudome Shinkan , Yamamoto Akio , Toyoshima Hideaki , Tamakoshi Akiko ,   for the JACC Study Group
Published: August 2005

To examine the possible association of dietary fat and fatty acids with breast cancer risk in a population with a low total fat intake and a high consumption of fish, we analyzed data from the Japan Collaborative Cohort (JACC) Study. From 1988 to 1990, 26 291 women aged 40–79 years completed a questionnaire on dietary and other factors. Intakes of fat or fatty acids were estimated by using a food frequency questionnaire. Rate ratios (RR) were computed by fitting proportional hazards models. During the mean follow‐up of 7.6 years, 129 breast cancer cases were documented. We found no clear association of total fat intake with breast cancer risk; the multivariate‐adjusted RR across quartiles were 1.00, 1.29, 0.95, and 0.80 (95% confidence interval [CI] 0.46–1.38). A significant decrease in the risk was detected for the highest quartile of intake compared with the lowest for fish fat and long‐chain n‐3 fatty acids; the RR were 0.56 (95% CI 0.33–0.94) and 0.50 (0.30–0.85), respectively. A decreasing trend in risk was also suggested with an increasing intake of saturated fatty acids (trendP = 0.066). Among postmenopausal women at baseline, the highest quartile of vegetable fat intake was associated with a 2.08‐fold increase in risk (95% CI 1.05–4.13). This prospective study did not support any increase in the risk of breast cancer associated with total or saturated fat intake, but it suggested the protective effects of the long‐chain n‐3 fatty acids that are abundant in fish. (Cancer Sci2005;96: 590 – 599)
A novel cinnamic acid derivative that inhibits Cdc25 dual‐specificity phosphatase activity
Authors: Aoyagi Yoshimi , Masuko Norio , Ohkubo Shuichi , Kitade Makoto , Nagai Kentaro , Okazaki Shinji , Wierzba Konstanty , Terada Tadafumi , Sugimoto Yoshikazu , Yamada Yuji
Published: August 2005

The Cdc25 dual‐specificity phosphatases are key regulators of cell cycle progression through activation of cyclin‐dependent kinases (Cdk). Three homologs exist in humans: Cdc25A, Cdc25B, and Cdc25C. Cdc25A and Cdc25B have oncogenic properties and are overexpressed in some types of tumors. Compounds that inhibit Cdc25 dual‐specificity phosphatase activity might thus be potent anticancer agents. We screened several hundred compounds in a library using an in vitro phosphatase assay, with colorimetric measurement of the conversion of p‐nitrophenyl phosphate (pNPP) to p‐nitrophenol by the catalytic domain of recombinant human Cdc25, and discovered TPY‐835, which inhibits Cdc25A and Cdc25B activity (IC50 = 5.1 and 5.7 µM, respectively). TPY‐835 had mixed inhibition kinetics for Cdc25A and Cdc25B. TPY‐835 caused cell cycle arrest in the G1 phase in human lung cancer cells (A549 and SBC‐5) but not cell cycle arrest in the G2/M phase. After treatment with TPY‐835, the activation of Cdk2 was suppressed and phosphorylation of the retinoblastoma (Rb) protein was decreased in SBC‐5 cells. In addition, TPY‐835 induced an increase of the sub‐G1 phase cell population after 48–72 h treatment. The growth inhibitory effects of TPY‐835 against cisplatin (CDDP)‐, camptothecin‐ and 5‐FU‐resistant cell lines are comparable to the growth inhibitory effect on their parental lines, thus indicating that TPY‐835 did not show cross‐resistance to these cell lines. These results suggest that TPY‐835 is a promising candidate for constructing a novel class of antitumor agents that can control the cell cycle progression of cancer cells. (Cancer Sci 2005; 96: 614–619)
Common signaling pathway is used by the trans‐interaction of Necl‐5/Tage4/PVR/CD155 and nectin, and of nectin and nectin during the formation of cell–cell adhesion
Authors: Sato Tatsuhiro , Irie Kenji , Okamoto Ryoko , Ooshio Takako , Fujita Naoyuki , Takai Yoshimi
Published: August 2005

Nectin is a Ca2+‐independent Ig‐like cell–cell adhesion molecule that forms homo‐ and hetero‐trans‐dimers (trans‐interaction). Nectin first forms cell–cell adhesions and then recruits cadherin to the nectin‐based cell–cell adhesion sites to form AJ cooperatively with cadherin. In addition, the trans‐interaction of nectin and nectin induces the activation of Cdc42 and Rac small G proteins, which enhances the formation of AJ. The activation of Cdc42 and Rac by the trans‐interaction of nectin and nectin is mediated by c‐Src, another small G protein, Rap1, a Cdc42‐GEF, FRG, and a Rac‐GEF, Vav2. Necl‐5/Tage4/PVR/CD155 is another Ca2+‐independent Ig‐like molecule, which does not homophilically trans‐interact, but heterophilically trans‐interacts with nectin‐3, one member of the nectin family. We show here that the trans‐interaction of Necl‐5 and nectin‐3 bidirectionally induces the activation of Cdc42 and Rac. Similarly to the activation of Cdc42 and Rac by the trans‐interaction of nectin and nectin, the trans‐interaction of Necl‐5 and nectin‐3 first recruits and activates c‐Src at the Necl‐5/nectin‐3‐based cell–cell contact sites. c‐Src then phosphorylates FRG and Vav2, and the tyrosine‐phosphorylated FRG and Vav2 are recruited to the Necl‐5/nectin‐3‐based cell–cell contact sites. The trans‐interaction of Necl‐5 and nectin‐3 also activates Rap1 through C3G, a Rap‐GEF, and this activation of Rap1 is required for the activation of Cdc42 and Rac. These results indicate that the trans‐interactions of Necl‐5 and nectin‐3 and of nectin and nectin induce the activation of Cdc42 and Rac through the common signaling molecules c‐Src, Rap1, FRG, and Vav2. (Cancer Sci 2005; 96: 578 –589)
Expression and function of LAT1, a neutral amino acid exchanger, in renal porcine epithelial cell line LLC‐PK1
Authors: Soares‐da‐Silva P. , Serrão P. , Fraga S. , Pinho M. J.
Published: August 2005

Aim:  The present study examined the expression of LAT1 and the functional characteristics of the inward and outward [14C] l‐leucine transporter in the renal porcine epithelial cell line LLC‐PK1.

Methods:  LLC‐PK1 cells were cultured in polycarbonate filters and accumulation and transepithelial flux of the substrate monitored with [14C] l‐leucine. LAT1 transcripts were examined by RT‐PCR. LAT1 protein was detected by immunoblotting.

Results:  The accumulation of [14C] l‐leucine in the cell and the [14C] l‐leucine transepithelial flux were four‐ and twofold, respectively, when the substrate was added from the basal cell side, suggesting that the basolateral membrane is endowed with a high density of transport units, when compared with the apical membrane. Increases in the transepithelial flux of [14C] l‐leucine by unlabelled l‐leucine were also more pronounced when unlabelled l‐leucine was added from the basolateral membrane. In the absence of Na+, unlabelled l‐leucine increased the basal and apical fractional outflow of [14C] l‐leucine, this being similar at pH 7.4 and pH 6.2. RT‐PCR and immunoblotting detected LAT1 transcript and protein, respectively.

Conclusion:  LLC‐PK1 cells are endowed with the LAT1 transcript and protein and transport l‐leucine through the Na+‐independent and pH‐insensitive LAT1 transporter. The density of transporter units in LLC‐PK1 cells may be higher at the basolateral membranes, although be also present in the apical membranes.
Platelet activating factor (PAF) increases plasma protein extravasation and induces lowering of interstitial fluid pressure (Pif) in rat skin
Authors: Iversen V. V. , Nedrebø T. , Borge B. A. , Salvesen G. S. , Reed R. K.
Published: August 2005

Aim:  To investigate the ability of the microdialysis technique to measure capillary selectivity of different sized plasma proteins induced by local administration of platelet activating factor (PAF).

Methods:  We used hollow plasmapheresis fibres with 3 cm membrane (cut off 3000 kDa) placed on the back of anaesthetized rats.

Results:  Platelet activating factor (50 μg mL−1) administered locally via the fibre, increased extravasation of radiolabelled 125I‐HSA from plasma to the microdialysis fibre by approximately 900% compared both to baseline and the control fibre within 70 min (n = 6, P < 0.05). The extravasation in the control fibre did not change over time. HPLC measurement of plasma proteins in the microdialysis perfusate also demonstrated decreased capillary selectivity for proteins in the diameter range of 73 Å, 56 Å and 39 Å after local administration of PAF (n = 6, P < 0.05). PAF also significantly lowered interstitial fluid (Pif) pressure after subcutaneous administration (50 μg mL−1). Mean arterial pressure (MAP) after intravenous injection of PAF (0.4 μg kg−1) fell instantly by about 50 mmHg, and stabilized at 50 mmHg after 15 min (n = 6). MAP was unaltered when PAF was given through the microdialysis fibre (n = 4). Both total tissue water (TTW) and extravasation of albumin, measured as the plasma‐to‐tissue clearance (E‐alb) showed a significant increase after PAF (n = 7, P < 0.05).

Conclusions:  The present study demonstrates that PAF induces plasma protein extravasation and decrease capillary selectivity of different sized plasma proteins. It also increases transcapillary fluid flux, and lowers Pif, indicating a role for PAF in the interstitium for generation of transcapillary transport of water and large molecules followed by formation of oedema.
Myocardial and coronary sinus purines as indicators of pig heart energy metabolism during reperfusion after extracorporeal circulation
Authors: Safranow K. , Rzeuski R. , Listewnik M. J. , Jakubowska K. , Rać M. E. , Olszewska M. , Chlubek D.
Published: August 2005

Aim  The precise understanding of myocardial metabolism is crucial for the optimization of cardiosurgical procedures. We attempted to gain a comprehensive insight into the purine metabolism of the porcine heart during reperfusion by measuring concentrations of nucleotides, nucleosides and oxypurines simultaneously in the myocardium and coronary sinus.

Methods  Twenty‐five pigs were subjected to sham cardiosurgery with extracorporeal circulation and cold cardioplegic arrest of 60 min. Myocardial biopsies, as well as coronary sinus and arterial blood samples were taken before aortic clamping and at 5, 20, 60 and 120 min of reperfusion. HPLC was used to measure concentrations of 17 purines in the bioptates and of 5 in plasma.

Results  Reperfusion rapidly normalized the ischaemic decrease in the adenylate energy charge of the myocardium, but during 120 min failed to restore the reduced adenylate pool, because of irreversible loss of nucleosides by cardiomyocytes. Low adenylate energy charge and depletion of the adenylate pool were accompanied by analogous changes in the guanylates and growing deficit of NAD and NADP. Reperfusion was marked by significant release of inosine and guanosine from the heart, without any noticeable effect on hypoxanthine and xanthine.

Conclusions  Coronary sinus concentrations of purines provide only a limited insight into the metabolism of the porcine heart. Repeated biopsies of the heart muscle and HPLC determinations of purine profiles represent a comprehensive and unique method for the study of purine metabolism during ischaemia and reperfusion. Future research on myocardial metabolism in disease and during cardiosurgical procedures should additionally be oriented to deficits in guanine and pyridine nucleotides.
Glycogen depletion in intrafusal fibres in rats during short‐duration high‐intensity treadmill running
Authors: Yoshimura A. , Toyoda Y. , Murakami T. , Yoshizato H. , Ando Y. , Fujitsuka N.
Published: August 2005

Aim:  The recruitment patterns of the intrafusal and extrafusal fibres in the soleus (SOL) and extensor digitorum longus (EDL) muscles of rats were investigated during brief‐intensity exercise by assaying their glycogen content histochemically.

Methods:  Six adult male rats were assigned to each of four groups that ran up a 6° incline on a motor‐driven treadmill, at 40 m min−1 for either 0.5, 1, 2, or 4 min. Six adult male rats in the control group did not run. Extrafusal and intrafusal fibres were classified by myosin ATPase staining. Optical densities for glycogen content were evaluated in serial periodic acid Schiff (PAS) stained‐sections from the B and C regions of intrafusal fibres.

Results:  The glycogen content of type IIA fibres in the SOL and EDL muscles decreased significantly in the early phase of exercise whereas the glycogen content of type I fibres in these muscles decreased later than that of type IIA fibres. The glycogen content of bag2 fibres decreased after 1 min of exercise in the SOL muscle and after 2 min of exercise in the EDL muscle. On the other hand, the glycogen content of bag1 and chain fibres decreased significantly after 2 min in the SOL muscle but not in the EDL muscle.

Conclusion:  The results suggest that during brief‐intensity exercise, as the glycogen content of type IIA fibres is reduced earlier than that of type I fibres, bag2 fibres are most important early in this type of exercise.
The effect of fluoxetine in patients with pain and constipation‐predominant irritable bowel syndrome: a double‐blind randomized‐controlled study
Authors: VAHEDI H. , MERAT S. , RASHIDIOON A. , GHODDOOSI A. , MALEKZADEH R.
Published: August 2005

Background : Irritable bowel syndrome has been treated with selective serotonin reuptake inhibitors but there is not enough evidence from controlled trials to prove their effectiveness.

Aim : To compare the effects of fluoxetine and placebo in the treatment of pain and constipation‐predominant irritable bowel syndrome in a double‐blind randomized‐controlled trial.

Methods : Forty‐four cases meeting Rome II criteria for irritable bowel syndrome with predominance of pain and constipation were included in this study. Organic causes were ruled out by detailed history, physical examination, laboratory tests and colonoscopy. Participants were then randomly assigned to receive either fluoxetine or placebo for 12 weeks. Symptoms addressed by the Rome II criteria were recorded during treatment and 4 weeks after termination of treatment.

Results : Fluoxetine was significantly more effective than placebo in decreasing abdominal discomfort, relieving feeling and sense of bloating, increasing frequency of bowel movements and decreasing consistency of stool. Mean number of symptoms per patient decreased from 4.6 to 0.7 in the fluoxetine group vs. 4.5 to 2.9 in controls (P < 0.001).

Conclusions : Fluoxetine is an effective and well‐tolerated short‐term treatment for pain and constipation‐predominant irritable bowel syndrome.
Direct and indirect costs in a prospective cohort of patients with pancreatic cancer
Authors: MÜLLER‐NORDHORN J. , BRÜGGENJÜRGEN B. , BÖHMIG M. , SELIM D. , REICH A. , NOESSELT L. , ROLL S. , WIEDENMANN B. , WILLICH S. N.
Published: August 2005

Background : Pancreatic cancer is an aggressive cancer with a low survival time. So far, there have been no studies assessing direct and indirect costs in individual patients.

Aim : To assess prospectively the cost of illness in patients with pancreatic cancer.

Methods : Patients were consecutively included at first admission to hospital. Sociodemographic factors, medical resource use and employment status were assessed by patient interviews and from medical records in a standardized way. Costs were calculated from the perspectives of the hospital, social insurance and society. Linear regression analyses were used to determine factors associated with costs.

Results : A total of 57 patients were admitted with suspected pancreatic cancer. Of these patients, 45 (79%) had pancreatic cancer as final diagnosis, 11 (19%) pancreatitis and one patient cystadenoma. The median survival was 10.9 months in patients with pancreatic cancer. Per month of observation from societal perspective, total costs were €4075 for patients. Costs of hospitalizations were responsible for 75% of total costs. In multivariable analyses, surgery, a lower educational level, younger age, and the prevalence of metastases were significantly associated with higher total costs.

Conclusions : Costs are considerable in patients with pancreatic cancer. Our results may facilitate further economic evaluations and aid health policy‐makers in resource allocation.
Clinico‐biochemical prediction of biliary cause of acute pancreatitis in the era of endoscopic ultrasonography
Authors: LIU C. L. , FAN S. T. , LO C. M. , TSO W. K. , WONG Y. , POON R. T. P. , LAM C. M. , WONG B. C. , WONG J.
Published: August 2005

Background : Occult biliary stones escape detection on conventional investigations, and clinico‐biochemical systems proposed for predicting biliary pancreatitis has low predictive values.

Aim : To evaluate the accuracy of clinico‐biochemical parameters for prediction of biliary pancreatitis in patients undergoing endoscopic ultrasonography.

Methods : Early endoscopic ultrasonography was performed on 139 patients presenting with acute pancreatitis within 24 h of admission. The aetiologies of all patients were determined after complete evaluations, and clinico‐biochemical characteristics of patients with a biliary cause (biliary group) and non‐biliary causes (non‐biliary group) were compared.

Results : Biliary pancreatitis was diagnosed in 107 patients and 32 patients had non‐biliary causes. The biliary group belonged to a significantly older age group, had a female predominance, significantly more derangement of liver function and a higher incidence of severe attack of acute pancreatitis. On multivariate analysis, female sex, age >58 years and serum alanine aminotransferase >150 U/L were independent predictive factors for biliary cause of acute pancreatitis. Using these three factors for prediction of biliary cause, the sensitivity was 93% and overall accuracy was 85%.

Conclusion : Clinico‐biochemical prediction for biliary cause of acute pancreatitis improves in the era of endoscopic ultrasonography with a higher sensitivity and overall accuracy. In centres where endoscopic ultrasonography is inaccessible or local expertise is unavailable, clinico‐biochemical prediction of biliary cause of acute pancreatitis may provide a useful alternative in the initial management of this group of patients.
On the limitation of 6‐tioguaninenucleotide monitoring during tioguanine treatment
Authors: DE BOER N. K. H. , DE GRAAF P. , WILHELM A. J. , MULDER C. J. J. , VAN BODEGRAVEN A. A.
Published: August 2005

Background : Tioguanine (thioguanine) has been proposed as a rescue thiopurine for azathioprine or mercaptopurine intolerant inflammatory bowel disease patients. The use of tioguanine leads to high 6‐tioguaninenucleotide (6‐thioguaninenucleotide) levels in red blood cells but, contra‐intuitively, these have yet not been associated with an increased risk of myelotoxicity.

Aim : To assess the role of 6‐tioguaninenucleotide concentrations in developing myelotoxicity during tioguanine treatment.

Methods : Database analysis of 25 patients treated with tioguanine. Clinical findings and laboratory parameters were related to 6‐tioguaninenucleotide levels.

Results : One patient developed a myelodepression (21 mg TG/day for 3 months and 6‐tioguaninenucleotide 714 pmol/8 × 108 red blood cells). 6‐Tioguaninenucleotide levels varied greatly between individuals (mean 6‐tioguaninenucleotide level 621 pmol/8 × 108 red blood cells, s.d. 340 pmol/8 × 108 red blood cells and range 34–1653 pmol/8 × 108 red blood cells). The TG dosages (mean 20.6 mg/day and median 20 mg/day) did not correlate with 6‐tioguaninenucleotide levels (r = 0.31, N.S.). High 6‐tioguaninenucleotide levels (>450 pmol/8 × 108 red blood cells) did not effect haemoglobin concentrations (mean 8 mmol/L), peripheral leucocyte (mean 7.5 × 109/L) or platelet counts (mean 298 × 109/L). No correlations were established between laboratory parameters, type of disease and 6‐tioguaninenucleotide level.

Conclusion : High 6‐tioguaninenucleotide levels in erythrocytes (>450 pmol/8 × 108) during TG treatment compared to azathioprine or mercaptopurine treatment are not indicative for (developing) myelotoxicity.
Allopurinol safely and effectively optimizes tioguanine metabolites in inflammatory bowel disease patients not responding to azathioprine and mercaptopurine
Authors: SPARROW M. P. , HANDE S. A. , FRIEDMAN S. , LIM W. C. , REDDY S. I. , CAO D. , HANAUER S. B.
Published: August 2005

Background : Many non‐responders to azathioprine or mercaptopurine (6‐mercaptopurine) have high normal thiopurine methyltransferase activity and preferentially metabolize mercaptopurine to produce 6‐methylmercaptopurine instead of the active 6‐tioguanine (6‐tioguanine) metabolites.

Aim : To describe the use of allopurinol in mercaptopurine/azathioprine non‐responders to deliberately shunt metabolism of mercaptopurine towards 6‐tioguanine.

Methods : Fifteen thiopurine non‐responders whose metabolites demonstrated preferential metabolism towards 6‐methylmercaptopurine are described. Subjects were commenced on allopurinol 100 mg po daily and mercaptopurine/azathioprine was reduced to 25–50% of the original dose. Patients were followed clinically and with serial 6‐tioguanine and 6‐methylmercaptopurine metabolite measurements.

Results : After initiating allopurinol, 6‐tioguanine levels increased from a mean of 185.73 ± 17.7 to 385.4 ± 41.5 pmol/8 × 108 red blood cells (P < 0.001), while 6‐methylmercaptopurine decreased from a mean of 10 380 ± 1245 to 1732 ± 502 pmol/8 × 108 RBCs (P < 0.001). Allopurinol led to a decrease in white blood cell from a mean of 8.28 ± 0.95 to 6.1 ± 0.82 × 108/L (P = 0.01).

Conclusions : The addition of allopurinol to thiopurine non‐responders with preferential shunting to 6‐methylmercaptopurine metabolites appears to be an effective means to shift metabolism towards 6‐tioguanine.
Fatigue in adult coeliac disease
Authors: SINISCALCHI M. , IOVINO P. , TORTORA R. , FORESTIERO S. , SOMMA A. , CAPUANO L. , FRANZESE M. D. , SABBATINI F. , CIACCI C.
Published: August 2005

Background : Fatigue is reported by many adults at the moment of diagnosis of coeliac disease and during follow‐up.

Aim : To evaluate the prevalence, characteristics and associations of fatigue in adult coeliac disease patients.

Methods : The investigated sample comprised adults from Campania, Italy. A total of 130 coeliac disease patients were consecutively recruited in both treated (59 on gluten‐free diet) and untreated conditions (71 on normal diet). The control group was made up of 80 healthy controls. Coeliac disease patients and healthy controls underwent laboratory tests, a set of questionnaires for studying fatigue: visual analogue scale for fatigue, chronic fatigue syndrome questionnaire, fatigue severity scale and a modified version of the Zung self‐rating depression scale.

Results : Coeliac disease patients showed a significantly lower body mass index than controls (P = 0.0001), lower serum iron (P = 0.04). The entire cohort of coeliac disease patients reported greater modified version of the Zung self‐rating depression scale score (P = 0.001), greater visual analogue scale for fatigue score (P = 0.0001) and greater chronic fatigue syndrome questionnaire score (P = 0.0001) compared with healthy controls. Coeliac disease patients on a gluten‐free diet had a significantly higher modified version of the Zung self‐rating depression scale score than coeliacs on a normal diet (P = 0.001). The prevalence of pathological modified version of the Zung self‐rating depression scale score was 17% in all coeliac disease patients and 0% in healthy controls. A significant correlation was found between modified version of the Zung self‐rating depression scale score and fatigue scale scores in coeliacs on a normal diet. Presence/absence of gastrointestinal symptoms did not show any significant correlation with modified version of the Zung self‐rating depression scale score and fatigue scale scores. In coeliacs on a gluten‐free diet, modified version of the Zung self‐rating depression scale and fatigue scales scores did not significantly differ from coeliacs on a normal diet and were not related to dietetic compliance.

Conclusion : In coeliacs, fatigue is a common finding, which ameliorates with the gluten‐free diet and is strictly correlated to depression although coeliacs on a gluten‐free diet showed more frequent and more severe depression symptoms than coeliacs on a normal diet.
Malic enzyme 1genotype is associated with backfat thickness and meat quality traits in pigs
Authors: Vidal O. , Varona L. , Oliver M. A. , Noguera J. L. , Sànchez A. , Amills M.
Published: August 2005

Malic enzyme 1 (ME1) is a part of the tricarboxylate shuttle that provides NADPH and acetyl‐CoA required in fatty acid biosynthesis. The pigME1locus maps on the proximal end of chromosome 1, where a quantitative trait loci (QTL) affecting fat deposition has been previously described. We amplified fragments of 1457 and 1459 bp that corresponded to the complete coding region and the 3′‐untranslated region (UTR), respectively, of the pigME1gene. The sequences of these two fragments in pigs from three breeds (Landrace, Large White and Piétrain) contained five single nucleotide polymorphisms (SNP) in the 3′‐UTR: C1706T, G1762T, A1807C, C1857A and T1880A. Three haplotypes were found in two generations of a selected Landrace population: H1 (C1706G1762A1807C1857A1880), H2 (C1706G1762A1807C1857T1880) and H3 (T1706T1762C1807A1857T1880). Using Bayesian association analyses, significant associations (highest posterior density at 95%) betweenME1genotype and backfat (BF) thickness at 171 days and muscular pH were found in a Landrace population.
The Evi1 proto‐oncoprotein blocks endomitosis in megakaryocytes by inhibiting sustained cyclin‐dependent kinase 2 catalytic activity
Authors: Kilbey Anna , Alzuherri Hadi , McColl Joan , Calés Carmela , Frampton Jonathan , Bartholomew Christopher
Published: August 2005

The 3q21q26 syndrome leukaemias are characterised by dystrophic megakaryocytes, elevated platelet counts, ectopic EVI1 protein production and poor prognosis. To investigate the molecular basis of this disease, we developed a model system to examine the biological activity of EVI1 in a megakaryocyte progenitor cell line. For this purpose, Evi1 was conditionally expressed in human erythroleukaemia cells (HEL) that progress along the megakaryocyte lineage in the presence of 12‐O‐tetradecanoylphorbol 13‐acetate (TPA). TPA‐stimulated HEL cells normally undergo: (1) growth arrest; (2) altered morphology; (3) endomitosis and (4) characteristic changes in gene expression, including reduction of the erythroid‐specific glycophoryn A and elevation of the specific glycoproteins GPIIIa and GPVI. Enforced Evi1 expression alone had no effect upon HEL cell proliferation or differentiation but a phenotype was manifest upon stimulation to differentiate. Evi1‐expressing, TPA‐treated HEL cells still showed growth arrest, had reduced and enhanced glycophoryn A and GPIIIa mRNA's, respectively, but failed to significantly elevate GPVI mRNA. This was accompanied by inhibition of endomitosis and altered cell morphology. Sustained CDK2 catalytic activity, typically associated with megakaryocyte endomitosis, was dramatically decreased in TPA‐stimulated Evi1‐expressing HEL cells because of significantly reduced levels of cyclin A. Therefore, enforced Evi1 expression could inhibit megakaryocyte differentiation although retention of some characteristic molecular changes, in combination with a block in endomitosis and altered morphology, suggest a defect in lineage progression. These results suggest that ectopic Evi1 expression contributes to a defective megakaryocyte differentiation programme and is likely to contribute to the phenotype observed in 3q21q26 syndrome leukaemias.
Allergic airway inflammation induces tachykinin peptides expression in vagal sensory neurons innervating mouse airways
Authors: Dinh Q. T. , Mingomataj E. , Quarcoo D. , Groneberg D. A. , Witt C. , Klapp B. F. , Braun A. , Fischer A.
Published: August 2005

Background
Allergic airway inflammation has been shown to induce pro‐inflammatory neuropeptides such as tachykinin peptides substance P (SP) and neurokinin A (NKA) together with related peptide like calcitonin gene‐related peptide (CGRP) in nodose sensory neurons innervating guinea‐pig airways.

Objective
The present study was designed to examine the effects of allergen sensitization and challenge on the SP/NKA expression in the jugular–nodose ganglion neurons innervating the murine airways.

Methods
Using retrograde neuronal tracing technique in combination with double‐labelling immunohistochemistry, the expression of SP/NKA was investigated in a murine model of allergic airway inflammation.

Results
Allergic airway inflammation was found to induce the expression of SP/NKA (13.2±1.43% vs. 5.8±0.37%, P<0.01) in large‐diameter (>20 μm) vagal sensory neurons retrograde labelled with Fast blue dye from the main stem bronchi.

Conclusion
Based on the induction of tachykinins in airway‐specific large‐sized jugular–nodose ganglia neurons by allergic airway inflammation, the present study suggests that allergen sensitization and challenge may lead to de novo induction of tachykinins in neurons. This may partly contribute to the pathogenesis of airways diseases such as allergic airway inflammation.
Plasminogen activator inhibitor type 1 (PAI‐1) in plasma and adipose tissue in HIV‐associated lipodystrophy syndrome. Implications of adipokines
Authors: He G. , Andersen O. , Haugaard S. B. , Lihn A. S. , Pedersen S. B. , Madsbad S. , Richelsen B.
Published: August 2005

Background  PAI‐1, an important inhibitor of fibrinolysis, is increased in obese subjects and has been shown to be an independent risk factor for cardiovascular disease. In the present study, we investigated the association between circulating levels of PAI‐1 and locally produced PAI‐1 in adipose tissue and body fat distribution and adipokines (TNF‐α, TNF receptors, IL‐6, IL‐8) in patients with and without HIV‐associated lipodystrophy syndrome (HALS).

Materials and methods  Eighteen men with HALS and 18 men with HIV but without HALS were investigated. DEXA and computed tomography scan were performed to determine total body fat and visceral adipose tissue mass. Insulin sensitivity was determined by the euglycaemic clamp technique. Plasma levels of PAI‐1 and cytokines were determined. In addition, PAI‐1, TNF‐α, IL‐6 and IL‐8 mRNA levels in subcutaneous adipose tissue were measured by real‐time reverse transcriptase polymerase chain reaction.

Results  HALS patients were characterized by a 3‐fold increased visceral adipose tissue (P < 0·001) and reduced limb fat (P < 0·01) as compared with non‐HALS patients but with no difference in total fat mass between the groups. Plasma PAI‐1 was increased in HALS patients (16·7 ng mL−1 vs. 8·2 ng mL−1, P < 0·05). Plasma PAI‐1 was positively correlated with BMI (r = 0·74, P < 0·01), plasma TNF‐α level (r = 0·64, P < 0·01), sTNFR‐I (r = 0·38, P < 0·05), and visceral fat (r = 0·67, P < 0·01). Moreover, plasma PAI‐1 was negatively associated with insulin sensitivity (r = −0·57, P < 0·01) and the percentage of limb fat (r = −0·57, P < 0·01). A positive correlation was found between plasma PAI‐1 and TNF‐α mRNA level. No association was, however, found between plasma PAI‐1 and PAI‐1 mRNA level in adipose tissue.

Conclusion  Plasma PAI‐1 is increased in HALS patients and it is suggested that dysregulation of the TNF‐system (high TNFα and high sTNFR1) may play a role in up‐regulating PAI‐1 in HALS.
Ischaemia or reperfusion: which is a main trigger for changes in nitric oxide mRNA synthases expression?
Authors: Pevni D. , Frolkis I. , Shapira I. , Schwartz D. , Schwartz I. F. , Chernichovski T. , Lev‐Ran O. , Sharony R. , Uretzky G.
Published: August 2005

Objective  To investigate alterations in endothelial nitric oxide synthase and inducible nitric oxide synthase mRNA expressions and nitric oxide release in the myocardium during ischaemia/reperfusion and determine whether these changes are ischaemic and/or reperfusion dependent.

Materials and methods  Isolated rat hearts were perfused by a modified Langendorff system. Following 1 h of global cardioplegic ischaemia, left ventricle haemodynamic parameters were recorded at baseline and during 30 min of reperfusion. Levels of endothelial, inducible nitric oxide synthases mRNA expression and nitric oxide release were measured at baseline, after ischaemia and at 30 min of reperfusion.

Results  Global cardioplegic ischaemia caused a significant depression of left ventricular function and a decrease of coronary flow. Postischaemic intensities of the endothelial nitric oxide synthase mRNA bands were significantly lower than at baseline (P < 0·01). There were no significant differences in endothelial nitric oxide synthase mRNA band intensities immediately after ischaemia compared to the end of reperfusion, nor between the intensities of inducible nitric oxide synthase mRNA bands at baseline, at end of ischaemia and at end of reperfusion. Nitric oxide in the myocardial effluent was below detectable levels at all measured points.

Conclusion  Ischaemic injury causes down‐regulation of endothelial nitric oxide synthase mRNA expression, which is then associated with reduction of coronary flow during reperfusion, representing one possible mechanism of ischaemia/reperfusion injury. We did not find expected elevations of inducible nitric oxide synthase mRNA expression during ischaemia or reperfusion and we suggest that ischaemia/reperfusion injury is not associated with nitric oxide overproduction.
Effect of soil aggregate size on infiltration and erosion characteristics
Authors: Abu‐Hamdeh N. H. , Abo‐Qudais S. A. , Othman A. M.
Published: August 2005

Detachment of soil particles by the processes of splash from rainfall and shallow flow from surface runoff is influenced by soil cohesion, soil aggregate properties, and characteristics of this flow. We have evaluated relationships between rates of detachment, aggregate size, and tensile strength of the soil. Soil and water losses were determined in the laboratory from sieved air‐dry samples on three aggregate size ranges of two clay loam soils differing in particle‐size distribution and organic matter. Tensile strength was measured for each aggregate size range. The results showed that as clod size increased, detachment rates increased and interaggregate tensile strength decreased. Wash erosion increased as initial clod size increased despite a decrease in runoff. Final rates of loss by splash were greater from the largest clods than from the smaller clods. Finally, splashed material was larger in size than material washed off. The fact that the size of the splashed material was larger than washed‐off material shows that material in the wash suffered more impact by raindrops and thus was more likely to be fragmented.
Identification of factor VIII gene mutations in 101 patients with haemophilia A: mutation analysis by inversion screening and multiplex PCR and CSGE and molecular modelling of 10 novel missense substitutions
Authors: Jayandharan G. , Shaji R. V. , Baidya S. , Nair S. C. , Chandy M. , Srivastava A.
Published: August 2005

Summary.  Haemophilia A (HA) is an X‐linked bleeding disorder caused by diverse mutations in the human coagulation factor VIII (FVIII) gene. We have analysed DNA from 109 unrelated Indian patients with HA for their FVIII gene defects. Among these patients 89 (82%) had severe (FVIII:C <1%) HA, 11 (10%) had moderate (FVIII:C 1–5%) HA and nine (8%) had mild (FVIII:C 5–30%) HA. These patients were first screened for the common intron 22 and intron 1 inversions. Inversion negative samples were screened for point mutations by a multiplex PCR and conformation sensitive gel electrophoresis strategy. Mutations were identified in 101 of the 109 patients. These included two (2%) intron 1 and 51 (51%) intron 22 inversions, four (4%) gross deletions and 44 (43%) point mutations. Twenty‐nine novel causative mutations, including 11 missense, seven frameshift, five nonsense mutations, three splice site defects and three gross deletions were detected. Ten of the novel missense mutations were studied by molecular modelling. Two different (Thr2253Pro and Pro1392fs) mutations were seen in four unrelated families and FVIII gene haplotyping suggested a common founder effect. Seven of these 109 patients had inhibitors. Among them, four had intron 22 inversions, one had a novel gross deletion (delexon 2–9) and one a nonsense mutation (Trp1535Stop). In one of these patients, no mutation could be identified in the FVIII gene. A Thr2253Pro novel mutation and an intron 22 inversion were identified in two female haemophiliacs. The data from this study suggests that the spectrum of gene defects in Indian patients with HA is as heterogeneous as reported in other populations.
Screening women with menorrhagia for underlying bleeding disorders: the utility of the platelet function analyser and bleeding time
Authors: Philipp C. S. , Miller C. H. , Faiz A. , Dilley A. , Michaels L. A. , Ayers C. , Bachmann G. , Dowling N. , Saidi P.
Published: August 2005

Summary.  Menorrhagia is a very common clinical problem among women of reproductive age and recent studies have suggested that underlying bleeding disorders, particularly von Willebrand's deficiency and platelet function defects, are prevalent in women presenting with menorrhagia. The objective of this study was to determine the utility of the platelet function analyser (PFA‐100) and bleeding time (BT) as initial screening tests for underlying bleeding disorders in women with menorrhagia. In this study, 81 women with a physician diagnosis of menorrhagia underwent PFA‐100 testing, BT and comprehensive haemostatic testing. The effectiveness of the PFA‐100 and BT as screening tools in women with menorrhagia was assessed using results of haemostatic testing for von Willebrand's disease (VWD) and platelet dysfunction. In women presenting with menorrhagia, the PFA‐100 had a sensitivity 80%, specificity 89%, positive predictive value (PPV) 33%, negative predictive value (NPV) 98% and efficiency 88% for VWD. For platelet aggregation defects, the PFA‐100 closure time had a sensitivity 23%, specificity 92%, PPV of 75%, NPV of 52% and efficiency 55%. The data suggest that the PFA‐100 may be useful in stratifying women with menorrhagia for further von Willebrand testing; however, neither the PFA‐100 nor the BT tests are effective for purposes of classifying women for standard platelet aggregometry testing in women presenting with menorrhagia.
Addressing current challenges in haemophilia care: consensus recommendations of a European Interdisciplinary Working Group
Authors: Ludlam C. A. , Mannucci P. M. , Powderly W. G. ,   The European Interdisciplinary Working Group
Published: August 2005

Summary.  Current challenges facing haemophilia care were identified and reviewed by an interdisciplinary group of experts in haemostasis and thrombosis, infectious disease, epidemiology, pharmacoeconomics and public health who met in February 2005 in Brussels. The outcome of this meeting was a series of consensus recommendations proposed to address the following three challenges: (i) developing the next generation of haemophilia specialists; (ii) reducing the risk that emerging pathogens present to safe haemophilia care and (iii) providing haemophilia care in an environment of cost constraint. It is intended that these consensus recommendations will form the basis of a concerted effort by leading haemophilia clinicians to secure future resources for the development and improvement of haemophilia care throughout Europe.