bluuebunny/crossref_metadata_2025 huggingface dataset:
Dietary Copper Deficiency Reduces the Elevation of Blood Pressure Caused by Nitric Oxide Synthase Inhibition in Rats
Authors: Saari Jack T.
Published: April 2003

Nitric oxide (NO)-mediated vasodilation is diminished in blood vessels of copper (Cu)-deficient rats. This study examined whether Cu deficiency affects blood pressure regulation via an effect on NO metabolism. Male, weanling rats were fed diets ranging from 0.4 to 7.2 mg Cu/kg diet for 5 weeks. Blood pressure was measured via arterial cannulation before and after intravenous injection of NG-nitro-L-arginine methyl ester (L-NAME, 10 mg/kg body weight), a NO synthase inhibitor. Plotting blood pressure against the liver Cu concentration (22–241 nmol/g) revealed that control mean arterial blood pressure (MAP; 104–152 mm Hg) was not correlated with the liver Cu concentration. However, both MAP after L-NAME treatment (MAP1, 148–190 mm Hg) and the elevation of MAP caused by L-NAME (ΔMAP; 8–68 mm Hg) were positively correlated with the liver Cu concentration (r = 0.52 for MAP1, r = 0.42 for ΔMAP; p < 0.05). This finding suggests that NO plays a significant role in maintaining basal blood pressure in Cu-adequate rats that is impaired by Cu deficiency.
Secular Trends in Pubertal Development
Authors: Karlberg Johan
Published: April 2003

Objective: To describe the secular trend in pubertal development in relation to the secular trend in height. Methods: Literature review of cross-sectional, longitudinal and twin studies. Results: Globally, there is a secular trend in adolescent growth for an increased mean final height at adulthood. To a lesser extent, there is also a secular trend towards earlier puberty. However, it seems that the two trends are not strongly connected. The increase in average height over the generations can, to a large extent, be explained by an earlier onset of the growth hormone-dependent phase of growth in early life. The age when this growth ‘spurt’ occurs in childhood strongly correlates with final height, but is not related to the age at peak height velocity during puberty. There is a large variation in the timing of the onset of puberty; it is largely influenced genetically, as implied by studies in twins. The single environmental factor that stands out as most significant – possibly explaining as much as 25% of the variation in the timing of puberty – is simply nutritional status in childhood; overnutrition and obesity seem to trigger pubertal onset. However, recent studies have identified that both shortness and thinness at birth are also associated with earlier pubertal maturation – a reverse of their impact during childhood years. Conclusions: More longitudinal studies are needed to understand the short- and long-term consequences of secular changes in both final height and pubertal development before we know how important the trends are.
LH and FSH Receptor Mutations and Their Effects on Puberty
Authors: Huhtaniemi Ilpo T.
Published: April 2003

Several mutations have been detected in the genes encoding the gonadotrophin receptors, and their phenotypic effects are observed in sexual differentiation, pubertal development and fertility of the affected individuals. Both activating and inactivating mutations are known for the gonadotrophin receptors. These mutations are rare, but they are very elucidating with respect to the details of gonadotrophin action, as well as clarifying the molecular pathogenesis of certain disorders in the development of reproductive functions. Proper diagnosis of these conditions by molecular biological techniques makes it possible to offer specific treatment for patients and proper counselling for patients and their families.
Puberty in Children with Cancer
Authors: Shalet Stephen M. , Brennan Bernadette M.D.
Published: April 2003

Cancer may impinge on puberty either directly through a mass lesion effect on the reproductive axis or indirectly through hormones secreted by tumours, for example human chorionic gonadotrophin, or weight loss, or the actual presence of a chronic disease process per se. The more frequent pubertal problems faced by children with cancer are due to the impact of treatment either on the central nervous system, the hypothalamic-pituitary axis or the gonad; in this review, we concentrate on these complications and their potential management.
Novel Treatment of Delayed Male Puberty with Aromatase Inhibitors
Authors: Dunkel Leo , Wickman Sanna
Published: April 2003

Background: As the evidence for the role of oestrogens in epiphyseal closure appears unequivocal, we hypothesized that boys with constitutional delay of puberty would attain greater adult height if oestrogen action was suppressed. Methods: We conducted a randomized, double-blind, placebo-controlled study in which we treated boys with constitutional delay of puberty with testosterone plus placebo or testosterone plus a potent fourth-generation aromatase inhibitor, letrozole. Findings: Letrozole effectively inhibited oestrogen synthesis. The 17β-oestradiol concentrations increased in the untreated group and in the testosterone/placebo-treated group, but in the testosterone/letrozole-treated group no such increase was observed until letrozole treatment was discontinued. Testosterone concentrations were threefold higher in the testosterone/letrozole-treated group than in the other groups. Within 18 months, bone age had advanced by 1.1 ± 0.3 years in the untreated group and by 1.7 ± 0.3 years in the testosterone/placebo-treated group, but only by 0.9 ± 0.2 years in the testosterone/letrozole-treated group (p = 0.02 between treatment groups). Predicted adult height did not change significantly in the untreated group and in the testosterone/placebo-treated group, whereas in the testosterone/letrozole-treated group the increase was 5.1 ± 1.2 cm (p = 0.004). Conclusions: Our findings suggest that, if oestrogen action is inhibited in growing adolescents, adult height will increase. This observation provides a rationale for studies aimed at delaying bone maturation in several growth disorders.
Recent Advances in Understanding Aminoglycoside Ototoxicity and Its Prevention
Authors: Wu Wei-Jing , Sha Su-Hua , Schacht Jochen
Published: April 2003

Studies over the last decade have left little doubt that reactive oxygen species (ROS) participate in the cellular events leading to aminoglycoside-induced hearing loss. The evidence ranges from the demonstration of aminoglycoside-mediated ROS formation in vitro to the prevention of ototoxicity by antioxidants in guinea pig in vivo. Here we review a hypothesis of the mechanism of toxicity, discuss possible causes underlying the gradient in base-to-apex sensitivity of outer hair cells, and present recent results on the adult mouse as a new animal model of aminoglycoside ototoxicity and its prevention.
Telomerase Activity and Its Correlation with the Proliferative Potential of Bone Marrow in Aplastic Anemia in Children
Authors: Shen JianBin , Tang JingYan , Zhao JinCai , Pan Ci , Chen Jing , Zhou Xin , Wang YaoPing
Published: April 2003

Objective: To explore telomerase activity and its association with the proliferative potential of hematopoietic stem cells in bone marrow (BM) in patients with aplastic anemia (AA). Methods: Telomerase activity of mononuclear cells separated from BM was determined with the TRAPezeTM kit. BM specimens from 22 cases with AA and 7 normal controls were included. SPSS10.0 was applied to analyze data derived from telomerase activity and colony-forming unit-granulocyte and monocyte. Results: The median telomerase activity level of BM in AA was 2-fold higher than that in normal controls. There was an inverse correlation in AA between telomerase activity and colony-forming unit (r = 0.78, p < 0.05). BM of chronic AA expressed higher telomerase than that of acute AA. Conclusions: Telomerase activity in AA was increased and may be the result of the negative feedback of hematopoietic potential. Telomerase activity varied between the subtypes of AA. Telomerase activity was conversely correlated with the proliferative potential of BM in AA.
Factors Influencing the Amount of Topical Preparations Applied
Authors: Matveev Nikolay V. , Maibach Howard I.
Published: April 2003

The studies on the application of topical preparations are reviewed. The data obtained demonstrate that in most cases the quantities of sunscreens, industrial protective creams and topical medications may differ greatly on different anatomic sites and may also vary due to numerous factors. The data on influencing factors in various studies were partially controversial, therefore a possible study design is suggested to resolve these controversies. It is also suggested that the standard layer thickness of sunscreens, used by standardization authorities to evaluate the sun-protective factor, should be reviewed, due to the evident failure of the majority of sunbathers to comply with the standard layer thickness used now by the ASA (2 mg/m2) or DIN (1.5 mg/m2). It is also important to have a standard of applied amounts of topical medications to be able to compare the effectiveness and bioequivalence of different topical medications precisely. The use of special delivery methods (such as flurandrenolide tape) demonstrates one approach to the standardization of topical treatment dosing.
Urinary Stone Formation: Dent’s Disease Moves Understanding Forward
Authors: Sayer John A. , Simmons Nicholas L.
Published: April 2003

Renal stones form in the late collecting duct in a complex milieu involving salts and protein components of the urine together with direct interactions at the epithelial cells lining the duct. The operation of newly discovered physiological controls that limit crystal formation by feedback mechanisms which sense the luminal environment are discussed. Adhesion at the epithelial surface and intracellular processing of crystals comprise a previously unrecognised mechanism for limiting crystal growth, which may be disrupted resulting in disease. Dent’s disease is discussed as a paradigm of a complex renal tubular disease resulting in renal stone formation. Defects in endosomal acidification, due to ablation of the CLC-5 voltage-gated Cl– channel, result in defects in both proximal and collecting duct endosomal traffic leading to stone formation.
Ionic Radiocontrast Media Disrupt Intercellular Contacts via an Extracellular Calcium-Independent Mechanism
Authors: Schick Cornelia Sonja , Bangert Renate , Kübler Wolfgang , Haller Christlieb
Published: April 2003

Direct cytotoxic effects of radiocontrast (RC) agents have been implicated in radiocontrast nephropathy (RCIN). The interaction between extracellular calcium, which plays a central role in intercellular contacts, and the in vitro toxicity of RC was tested in Madin-Darby canine kidney (MDCK) cell monolayers grown on permeable supports. Cell viability was determined by trypan blue exclusion. The function of intercellular junctions was assessed by measuring the electrical transmonolayer resistance (TMR). The cell contacts were examined with indirect immunofluorescence microscopy using antibodies against the junctional proteins E-cadherin, ZO-1 and occludin. The ionic RC agents diatrizoate and ioxaglate (74 mg iodine/ml), but not the nonionic compounds iohexol or iodixanol, decreased ionized calcium (Ca2+) in the incubation media from 1.48 ± 0.04 mM (control) to 0.89 ± 0.06 mM (diatrizoate), respectively to 1.05 ± 0.08 mM (ioxaglate). Diatrizoate, and to a lesser extent ioxaglate, reduced the number of viable MDCK cells and showed a redistribution of the E-cadherin, ZO-1 and occludin immunofluorescence signal with a parallel decrease of the TMR indicating an impaired monolayer integrity. A similar reduction of extracellular Ca2+ through EGTA failed to reproduce these effects. Conversely, raising Ca2+ in diatrizoate-containing media to control levels did not abrogate its toxicity. In conclusion, the ionic RC agents diatrizoate and ioxaglate, but not the nonionic compounds iohexol or iodixanol, reduce extracellular Ca2+ in vitro. However, this reduction of Ca2+ does not explain their cytotoxic effects which could contribute to the pathogenesis of RCIN in vivo by opening intercellular junctions.
The Plasma Homocysteine Levels Are Increased in Polycystic Ovary Syndrome
Authors: Loverro Giuseppe , Lorusso Filomenamila , Mei Loredana , Depalo Raffaella , Cormio Gennaro , Selvaggi Luigi
Published: April 2003

Women with polycystic ovary syndrome (PCOS) have several cardiovascular disease risk factors. Since hyperhomocysteinemia is associated with early atherosclerosis, it was postulated that the homocysteine levels are higher in PCOS patients than in control subjects which, therefore, may explain the cardiovascular disease risk. Thirty-five women with PCOS and 20 healthy subjects were studied. Endocrine assays, lipid profile, homocysteine and insulin level determinations, and ultrasound evaluation were performed in all subjects. We found significantly higher mean plasma homocysteine concentrations in patients with PCOS as compared with controls (10.4 ± 4.4 vs. 7.2 ± 1.5 ng/dl; p < 0.003). These data show that in PCOS early atherosclerosis is not exclusively dependent on hyperinsulinemia and elevated lipid profile – PCOS patients are exposed to significantly higher homocysteine levels which might increase the cardiovascular disease risk.
Cortical-Sparing Laparoscopic Adrenalectomy in a Patient with Multiple Endocrine Neoplasia Type IIA
Authors: Porpiglia F. , Destefanis P. , Bovio S. , Allasino B. , Orlandi F. , Fontana D. , Angeli A. , Terzolo M.
Published: April 2003

We describe the case of a patient affected by multiple endocrine neoplasia type IIA with a new diagnosis of an asymptomatic right pheochromocytoma. The patient underwent laparoscopic adrenalectomy with adrenal sparing. The removal of the tumor was successful with preservation of about one third of the adrenal gland. At the time of the last follow-up, the patient is well with partial hypoadrenalism without replacement therapy. The limitations to cortical-sparing adrenalectomy imposed by traditional open surgery (small tumor with peripheral location) can be reconsidered using the laparoscopic approach. Laparoscopic cortical-sparing adrenalectomy should become the gold standard for treatment of bilateral pheochromocytoma. The advantages of this technique are its efficacy and its reduced invasiveness with a low rate of complications either during the operation or in the postoperative period. Moreover, the preservation of a portion of the adrenal cortex may prevent the need for a life-long steroid replacement therapy.
Inhibin B Levels in Adolescents and Young Adults with Type 1 Diabetes
Authors: Salardi Silvana , Zucchini Stefano , Cicognani Alessandro , Gualandi Stefano , Barbieri Eveline , Cacciari Emanuele
Published: April 2003

Objective/Methods: To assess exocrine and endocrine testicular function in subjects with diabetes, we evaluated serum inhibin B, gonadotrophins and testosterone levels in 33 male adolescent and young adult patients affected by type-1 diabetes (age 21.0 ± 5 years; range 14.2–33.3), with a mean disease duration of 12.7 ± 5.8 years (range 1.5–25.3) and various metabolic control (HbA1c 7.8 ± 1.5%; range 5.5–13.2) and compared them with those of an age-matched group of 36 healthy control subjects (age 19.5 ± 4.1 years; range 13.6–28.1). Both patients and controls had a testicular volume ≧15 ml. Inhibin B was measured by ELISA method. Results/Conclusion: Diabetics and controls had comparable inhibin B (203 ± 74 vs. 221 ± 69 pg/ml, respectively) and follicle-stimulating hormone (FSH) levels, while luteinizing hormone (LH) and testosterone levels were significantly higher in the diabetic group. Inhibin B was negatively correlated both in patients and controls with FSH, while a negative correlation with LH was found only in the diabetic group. We conclude that our young diabetic males, after a mean disease duration of 12 years and various metabolic control, had inhibin B and FSH levels comparable to those of normal subjects. Therefore, they seem to have a regular testicular function and in particular a normal seminiferous tubule/Sertoli cell activity despite sustained hyperglycemia.
Experimental Split Cord Malformations
Authors: Emura Takaki , Asashima Makoto , Furue Miho , Hashizume Kohei
Published: April 2003

Objective: To induce experimental split cord malformations (SCMs) produced through the surgical induction of a dorsal midline fistula. Methods: In addition, the theory of embryogenesis of SCMs was verified by examining the developmental process of this experimentally induced anomaly. In Cynopus pyrrhogaster (amphibian) embryos (stage 18), the neural plate and notochord were split regionally to construct a fistula that appeared to be the ectopic neurenteric canal. Following this procedure, the embryonic development was traced morphologically and histologically. Results: Following the incubation and breeding period, split cord malformation was observed in some animals. Scoliosis, spina bifida, vertebral anomaly and subcutaneous manifestations were also observed with SCMs. Conclusions: The observations made in these experimentally induced SCMs are consistent with the findings in human SCMs. We report an experimental animal model of split cord malformation, in which double spinal cords were developed in the spinal canal. In addition, we examined the embryogenesis of SCMs. This study indicates that SCMs may arise through a process of dorsal midline fistula of the neural plate.
Duration of Antibiotic Therapy for the Treatment of Shunt Infection: A Surgeon and Patient Survey
Authors: Arthur Adam S. , Whitehead William E. , Kestle John R.W.
Published: April 2003

The ideal duration of antibiotic treatment for shunt infection remains a major unanswered question in pediatric neurosurgery. To date, no study has objectively determined the best length of treatment, i.e. that which minimizes both the length of hospital stay and the chance of reinfection. This study was undertaken to determine whether an increase in reinfection risk would be tolerated if the duration of therapy were shortened. Sixty-one members of the American Society of Pediatric Neurosurgeons (44 responding) and 831 patients or parents of patients with shunts (385 responding) were surveyed using similar questionnaires. Sixty-four percent of neurosurgeons (28/44) and 54% of patients (178/325) responding to the survey would tolerate an increased reinfection risk in order to gain a shorter duration of treatment. These data support the feasibility of a planned randomized study to determine the ideal length of antibiotic treatment for shunt infection.
Significance of Vitamin D Receptor Gene Polymorphism for Risk and Disease Severity of Prostate Cancer and Benign Prostatic Hyperplasia in Japanese
Authors: Hamasaki T. , Inatomi H. , Katoh T. , Ikuyama T. , Matsumoto T.
Published: April 2003

Objective: Recent studies have demonstrated an association between vitamin D receptor (VDR) genotype and prostate cancer. Currently, there is a scarcity of data regarding the association of VDR genotype with benign prostatic hyperplasia (BPH). The purpose of this study was to investigate the TaqI VDR polymorphism in Japanese prostate cancer patients, Japanese BPH patients and Japanese controls in order to determine if an association exists between VDR genotype and the risk of developing prostate cancer and BPH as well as disease severity. Methods: 110 prostate cancer patients, 83 BPH patients and 90 male age-matched controls were genotyped for a previously described TaqI restriction fragment length polymorphism at codon 352 in exon 9 of the VDR gene. Products were digested into T allele or t allele according to the absence or presence of a TaqI restriction site with individuals being classified as TT, Tt or tt. Results: The frequency of the genotype tt was higher in the control group (6.7%) compared to patients with prostate cancer (1.8%) and BPH (3.6%) but this was not statistically significant. However, the frequency of the genotype TT was significantly higher among prostate cancer patients with locally advanced or metastatic disease (T3/ T4/N1/M1) compared to controls (p = 0.001). In addition, the genotype TT was significantly higher among prostate cancer patients with a high Gleason grade of tumor (grade 5) compared to controls (p = 0.0001). In addition, the genotype TT was statistically higher in BPH patients with high prostate volume (volume >50 cm3) compared to controls (p = 0.001). Conclusion: These data demonstrate that VDR genotype plays an important role in determining the risk of more advanced and aggressive prostate cancer as well as prostatic enlargement in Japanese men.
The Effect of Diltiazem on the Healing of Traumatic Urethral Inflammation
Authors: Dündar Mehmet , Koçak Izzet , Erkus Muhan , Erol Haluk
Published: April 2003

Objective: To investigate the effect of diltiazem, a calcium channel blocker, on healing of the traumatic urethral inflammation when applied systemically or locally. Materials and Methods: 21 adult male Wistar rats (230–250 g) were assigned to group 1 (n = 7) = control, group 2 (n = 7) = local application or group 3 (n = 7) = systemic application. In group 1, only a urethral injury was achieved at the 12-o’clock position by gently introducing and drawing a tiny hook in the urethra until urethral bleeding occurred. In group 2, after the same procedure, 10 mg/kg diltiazem was applied retrogradely via a 22-gauge Angiocath intraurethrally for 5 consecutive days, while the same account of the drug was administered intraperitoneally in group 3. After 21 days, the rats were sacrificed for urethrectomy. Pathologically, the thickness of connective tissue, the regularity of the epithelial lining, the presence of the inflammation and the density of collagen were evaluated with Masson’s trichrome staining. The Mann-Whitney U test was used for statistical analyses. Results: The mean connective tissue thickness was 0.77 ± 0.39, 1.01 ± 0.77 and 0.93 ± 0.53 µm in groups 1, 2 and 3, respectively. The differences between the groups were insignificant (p > 0.05). The hyperplastic epithelial lining in the study groups, with both systemic and local applications, was markedly infrequent and the inflammation was less prominent. However, these differences did not reach statistical significance. Conclusions: Diltiazem appears not to have any preventive effect on connective tissue formation when applied locally or systemically in our urethral injury model.
Treatment of Advanced Hepatocellular Carcinoma with Biweekly High-Dose Gemcitabine
Authors: Ulrich-Pur Herbert , Kornek Gabriela Verena , Fiebiger Wolfgang , Schüll Birgit , Raderer Markus , Scheithauer Werner
Published: April 2003

Introduction: Until today, an optimal palliative treatment regimen has not been defined for patients with advanced hepatocellular carcinoma. Since the novel cytidine analog gemcitabine has shown strong antitumor effects in vitro in a human hepatoma cell line and its therapeutic potential seems well established in several different tumors including gastrointestinal adenocarcinomas, the present phase II trial using a dose-intensified biweekly administration schedule was initiated. Patients and Methods: 17 patients with histologically confirmed unresectable advanced or metastatic hepatoma were treated with gemcitabine 2,200 mg/m2 given as a 30-min intravenous infusion on days 1 and 15. Treatment courses were repeated every 4 weeks. Results: All patients were evaluable for response and toxicity assessment. No objective response was achieved, stable disease occurred in 8 patients (47%), and 9 progressed while on chemotherapy. The median time to progression was 4 months (range 1.5–14 months), and the median survival time was 8.5 months (range 2.5–16.0+ months). Treatment was well tolerated with mild or moderate leukopenia, thrombocytopenia and anemia representing the most common side effects. Gastrointestinal and other subjective toxicities were infrequent and also generally mild. Conclusions: In view of the disappointing treatment results, gemcitabine using this particular dose regimen should not be considered for further investigation in patients with advanced hepatocellular carcinoma.
Transmission of Mutans Streptococci in Adults within a Chinese Population
Authors: Nie Min , Fan Mingwen , Bian Zhuan
Published: April 2003

The purpose of this study was to investigate whether there was transmission between adults in Chinese families using chromosomal DNA fingerprinting. Plaque samples were obtained from buccal and occlusal surfaces of 11 married couples. The colonization levels of mutans streptococci were estimated as colony-forming units per milliliter, and caries experience was scored by decayed, missing and filled teeth. Information about medical history, diet regimes and age at marriage was obtained. The isolates were serotyped by biochemical test and genotyped using the restriction endonuclease HaeIII. The procedure was repeated after 3 months. The results showed that 1 couple had the same genotype of mutans streptococcus at the first examination, but this could not be repeated for the husband who had lost his mutans streptococci at the second examination. On the contrary, another couple that did not have the same mutans streptococcal genotype at the beginning had the same genotype after 3 months. No matching of genotypes was observed within 8 couples. In 1 male, no mutans streptococci were detected, therefore that couple was not considered. These data indicate that spouses had a chance to be infected by strains of mutans streptococci from another person. The results suggest that there may be transmission between adults in Chinese families, but it may be difficult for mutans streptococci to colonize another mouth permanently.
Molecular cytogenetic analysis consistently identifies translocations involving chromosomes 1, 2 and 15 in five embryonal rhabdomyosarcoma cell lines and a PAX-FOXO1A fusion gene negative alveolar rhabdomyosarcoma cell line
Authors: Roberts I. , Gordon A. , Wang R. , Pritchard-Jones K. , Shipley J. , Coleman N.
Published: April 2003

Rhabdomyosarcoma in children is a “small round blue cell tumour” that displays skeletal muscle differentiation. Two main histological variants are recognised, alveolar (ARMS) and embryonal (ERMS) rhabdomyosarcoma. Whereas consistent chromosome translocations characteristic of ARMS have been reported, no such cytogenetic abnormality has yet been described in ERMS. We have used multiple colour chromosome painting to obtain composite karyotypes for five ERMS cell lines and one PAX-FOXO1A fusion gene negative ARMS. The cell lines were assessed by spectral karyotyping (SKY), tailored multi-fluorophore fluorescence in situ hybridisation (M-FISH) using series of seven colour paint sets generated to examine specific abnormalities, and comparative genomic hybridisation (CGH). This approach enabled us to obtain karyotypes of the cell lines in greater detail than previously possible. Several recurring cytogenetic abnormalities were demonstrated, including translocations involving chromosomes 1 and 15 and chromosomes 2 and 15, in 4/6 and 2/6 cell lines respectively. All six cell lines demonstrated abnormalities of chromosome 15. Translocations between chromosomes 1 and 15 have previously been recorded in two primary cases of ERMS by conventional cytogenetics. Analysis of the translocation breakpoints may suggest mechanisms of ERMS tumourigenesis and may enable the development of novel approaches to the clinical management of this tumour.
Conservation of RET proto-oncogene splicing variants and implications for RET isoform function
Authors: Carter M.T. , Yome J.L. , Marcil M.N. , Martin C.A. , Vanhorne J.B. , Mulligan L.M.
Published: April 2003

The RET proto-oncogene encodes a receptor tyrosine kinase required for development of the kidney and neural crest-derived cell types. Alternative splicing of the 3′ exons of human RET results in three protein isoforms with distinct C-termini: RET9, RET51, and RET43. These RET isoforms show differential binding to downstream adapter molecules, suggesting they may have distinct signaling functions. We have characterized Ret 3′ sequences in mouse and investigated alternative splicing of this region. We found that the organization of Ret 3′ sequences is very similar to human RET. The mouse locus also has alternatively spliced C-terminal coding regions, and the sequences corresponding to RET9 and RET51 are highly conserved in both position and sequence with the human locus. Further, we compared the predicted C-terminal amino acids of RET9 and RET51 in seven vertebrate species, and found that they are well conserved. We have identified sequence encoding a putative ret43 isoform in mouse, however the predicted amino acid sequence showed low homology to human RET43. Our data suggest that RET isoforms are evolutionarily highly conserved over a broad range of species, which may indicate that each isoform has a distinct role in normal RET function.
Comparative mapping of five coding DNA sequences on cattle chromosomes 7 and 25
Authors: Goldammer T. , Kata S.R. , Brunner R.M. , Schwerin M. , Womack J.E.
Published: April 2003

Comparative mapping of four genes and one unknown coding DNA sequence in breakpoint positions of bovine chromosomes (BTA) 7 and 25 are presented. Performing a genome data base search five bovine expressed sequence tags from the MARC library matched with human genes coding for the general transcription factor IIIC polypeptide 1 (GTF3C1), the hypothetical protein KIAA0556, the interleukin 4 receptor (IL4R), the regulatory factor X-associated ankyrin-containing protein (RFXANK), and with an unknown human coding sequence partially homologous to the genomic cosmid clone R30923. Loci for these sequences were COMPASS predicted on BTA7 or BTA18 and to BTA18 or BTA25. Mapping was performed in a cattle-hamster somatic hybrid cell panel and a cattle-hamster 5000 rad whole genome radiation hybrid panel. GTF3C1, KIAA0556 and IL4R were assigned to the centromere region of BTA25 and RFXANK and R30923 close to the centromere of BTA7. The assignments contribute to the identification of evolutionary chromosome break points between human chromosomes 16 and 19 and BTA7, BTA18, and BTA25.
The mouse Arhgef6 gene: cDNA sequence, expression analysis, and chromosome assignment
Authors: Kutsche K. , Gal A.
Published: April 2003

Mutations in ARHGEF6, encoding a guanine nucleotide exchange factor for Rho GTPases, have been shown to cause X-chromosomal non-specific mental retardation (MRX) in human. Here we report the identification and characterization of the orthologous mouse gene, Arhgef6. The gene transcript of approximately 4.4 kb is present in various mouse tissues indicating ubiquitous expression as found for human ARHGEF6. The open reading frame encodes a polypeptide of 771 amino acids with high homology to human ARHGEF6. The structural motifs of both proteins are conserved including an N-terminal CH domain, followed by an SH3 domain, and a tandem organization of the DH and PH domains. Analysis of the temporal expression pattern revealed that Arhgef6 is strongly expressed in the very early embryo and somewhat less in later stages. A genomic cosmid clone containing Arhgef6 exons 1 and 2 has been isolated and used for mapping the gene on the mouse X chromosome.
Chorionic Villus Sampling: A 15-Year Experience
Authors: Papp Csaba , Beke Artúr , Mezei Gábor , Tóth-Pál Ernõ , Papp Zoltán
Published: April 2003

The authors describe experiences gained over the period of 1984–1999 at two medical centers with chorionic villus sampling (CVS). Altogether 1,149 CVSs had been performed between the 10th and 32nd gestational weeks. Prior to 1993 the transcervical approach (TC-CVS), after 1994 the transabdominal method (TA-CVS) was used. Analysis of data collected within the framework of this study was based on the following factors: indications for sampling, complications and incidence of pregnancy loss. 91.6% of the CVSs were carried out for the purposes of cytogenetic examination of the fetus. Over the past few years an increasing number of procedures had been carried out for molecular-genetic tests (7.6% of the total number of cases). Though the primary indication for cytogenetic tests was the advanced age of the mother, a remarkable increase in the number of samplings had taken place for the purpose of examining ‘suspicious ultrasound findings’, minor anomalies detected by ultrasound. In this group the proportion of pathological cases was significantly higher (14%) than in all the other samplings, carried out for other indications. This data in itself underlines the importance of ultrasound screening performed in the 18–20th weeks of gestation. Over the first half of the period being reviewed (1984–1993, TC-CVS), a fetal loss of 4.8% occurring within 3 weeks from the date of sampling, dropped to 1.7% in the period subsequent to year 1994 (TA-CVS). In cases of TA-CSV, both the complications and spontaneous abortions were fewer. In 74.1% of the cases studied, birth had taken place after the 37th week of gestation. Premature births (6.4%) and stillbirth rate (1.1%) did not exceed normal rates observed in the general population. On the basis of our results, it is safe to say that in prenatal diagnosis, TA-CVS is a real alternative method of mid-trimester amniocentesis and it is recommended for use at any stage of the pregnancy.
Congenital Bowing of Long Bones: Prenatal Ultrasound Findings and Diagnostic Dilemmas
Authors: Farra Chantal , Piquet Caroline , Guillaume Marc , D’Ercole Claude , Philip Nicole
Published: April 2003

Objective: Bowing of the long bones can be easily detected on antenatal ultrasound screening, but it is a nonspecific sign that can be associated with a variety of conditions, each denoting a different prognosis. Among these conditions, three well-differentiated bone dysplasias share bowed long bones as a main clinical manifestation. Campomelic dysplasia and Stüve-Wiedemann syndrome are characterized by a poor prognosis. Conversely, the overall prognosis of children affected with kyphomelic dysplasia is good, the intelligence and motor development are normal and the radiological abnormalities tend to improve and regress with age. Case Report: We report a case of prenatal detection of short and bowed femora at the 22nd week of gestation. Careful US examination as well as in utero X-ray of the skeleton allowed the exclusion of campomelic dysplasia. In the absence of an unambiguous diagnosis, the pregnancy was continued. At birth, the child presented with clinical and radiological features consistent with a diagnosis of kyphomelic dysplasia. Conclusion: This case illustrates the difficulties in making an accurate diagnosis and consequently giving a prognosis when isolated femoral bowing is found on fetal ultrasound examination.
Identification and Characterisation of the IgE-Binding Proteins 2S Albumin and Conglutin γ in Almond (Prunus dulcis) Seeds
Authors: Poltronieri P. , Cappello M.S. , Dohmae N. , Conti A. , Fortunato D. , Pastorello E.A. , Ortolani C. , Zacheo G.
Published: April 2003

Background: Almond proteins can cause severe anaphylactic reactions in susceptible individuals. The aim of this study was the identification of IgE-binding proteins in almonds and the characterisation of these proteins by N-terminal sequencing. Methods: Five sera were selected from individuals with a positive reaction to food challenge. Sodium dodecylsulphate-polyacrylamide gel electrophoresis and immunoblotting were performed on almond seed proteins. Purified IgE-binding proteins were tested for immunoblot inhibition with sera pre-incubated with extracts of hazelnut and walnut. Results: N-terminal sequences of the 12-, 30- and 45-kD proteins were obtained. The 45- and 30-kD proteins shared the same N terminus, with 60% homology to the conglutin γ heavy chain from lupine seed (Lupinus albus) and to basic 7S globulin from soybean (Glycine max). The sequences of the N-terminal 12-kD protein and of an internal peptide obtained by endoproteinase digestion showed good homology to 2S albumin from English walnut (Jug r 1). Immunoblot inhibition experiments were performed and IgE binding to almond 2S albumin and conglutin γ was detected in the presence of cross-reacting walnut or hazelnut antigens. Conclusions: Two IgE-binding almond proteins were N-terminally sequenced and identified as almond 2S albumin and conglutin γ. Localisation and conservation of IgE binding in a 6-kD peptide obtained by endoproteinase digestion of 2S albumin was shown.
Theophylline Inhibits NF-κB Activation in Human Peripheral Blood Mononuclear Cells
Authors: Umeda Masashi , Ichiyama Takashi , Hasegawa Shunji , Kaneko Miho , Matsubara Tomoyo , Furukawa Susumu
Published: April 2003

Background: Theophylline not only dilates the bronchi, but also modulates the production of proinflammatory cytokines and inhibits inflammation. Theophylline exerts an antiinflammatory effect on allergic inflammation through inhibition of NF-ĸB activation in mast cells. However, the action of theophylline on monocytes/macrophages and T cells is unknown. Methods: We examined whether or not theophylline inhibits tumor necrosis factor (TNF)-α-induced activation of the nuclear transcription factor NF-ĸB, a factor that is essential for the expression of proinflammatory cytokines, in human monocytic U-937 cells, a T cell line (Jurkat) and peripheral blood mononuclear cells (PBMC). The inhibitory effect of theophylline on TNF-α-induced NF-ĸB activation was evaluated by Western blotting, flow cytometry and chloramphenicol acetyltransferase (CAT) assaying. Expression of the IĸBα protein was evaluated by Western blotting. Results: Western blotting demonstrated that theophylline inhibits NF-ĸB activation in U-937 and Jurkat cells and PBMC. Flow cytometry demonstrated that theophylline inhibits NF-ĸB activation in U-937 and Jurkat cells in a dose-related manner. CAT assaying indicated that NF-ĸB-dependent reporter gene expression is inhibited in U-937 cells pretreated with theophylline. Western blotting of cytoplasmic extracts of U-937 cells revealed that this inhibition was linked to theophylline-induced preservation of expression of the IĸBα protein. Conclusions: These findings are consistent with the idea that theophylline suppresses the production of proinflammatory cytokines via inhibition of NF-ĸB activation through preservation of the IĸBα protein in monocytes/macrophages and T cells.
Effect of Extracellular Matrix Proteins on Platelet-Activating Factor-Induced Eosinophil Chemotaxis
Authors: Yoshikawa Mamoru , Matsumoto Kenji , Iida Makoto , Akasawa Akira , Moriyama Hiroshi , Saito Hirohisa
Published: April 2003

Background: Eosinophils accumulate in tissues during various allergic inflammatory processes. Selective eosinophil recruitment is the result of orchestrated events involving cell adhesion molecules and chemoattractants. However, the exact mechanism of the regulation of integrin avidity during interstitial migration is poorly understood. Objective: The purpose of this study was to investigate whether extracellular matrix proteins might activate human eosinophil chemotaxis and, if so, to clarify the mechanism in terms of integrin avidity. Method: Eosinophils were purified from the peripheral blood of healthy donors. Eosinophil migration was measured using Boyden chambers with filter membranes coated with fibronectin (Fn), vitronectin (Vn), laminin (Ln), hyaluronic acid, collagen type IV, or bovine serum albumin (BSA) overnight. Platelet-activating factor (PAF) was introduced into the lower chamber of each well. Eosinophils were placed in the upper chamber after incubation with IL-5 for 15 min. The number of eosinophils that transmigrated into the lower chamber was calculated by measuring the eosinophil peroxidase activity. Results: Fn, Ln and BSA enhanced PAF-induced chemotaxis of eosinophils. Inhibition experiments using blocking monoclonal antibodies showed that in the early phase of chemotaxis, Fn and Ln facilitated eosinophil chemotaxis that was mediated by α4 and α6 integrins, respectively. In the late phase of chemotaxis, BSA, but not other matrix proteins, facilitated both chemokinesis and chemotaxis that was mediated by β2 integrin. Conclusion: Our data strongly suggest that during chemotaxis, matrix proteins might activate eosinophils via binding with integrins to facilitate PAF-induced chemotaxis, and that such a mechanism might participate in allergic inflammatory processes.
Phase II Study of Uracil-Tegafur in Patients with Metastatic Pancreatic Cancer
Authors: Ueno Hideki , Okada Shuichi , Okusaka Takuji , Ikeda Masafumi , Kuriyama Hitoshi
Published: April 2003

Objective: Uracil-tegafur (UFT) has been reported to have a broad anti-tumor activity in a variety of malignancies including colorectal cancer and breast cancer. However, its activity in pancreatic cancer has not been fully evaluated. The aim of the present study was to evaluate the anti-tumor activity and toxicity of UFT in patients with metastatic pancreatic cancer. Methods: All patients were required to have a pathologic diagnosis of pancreatic adenocarcinoma with measurable metastatic lesions, and no prior chemotherapy. A dose of 360 mg/m2/day of UFT was administered orally until the appearance of disease progression or unacceptable toxicity. Results: Twenty-two patients were entered into this study. Of 21 patients evaluable for response, no patient achieved an objective tumor response; one showed no change, and the remaining 20 showed progressive disease. The median survival time for all patients was 4.2 (range: 0.9–9.0) months. The most common toxicities were nausea/vomiting and anorexia. Five patients (23%) had to discontinue UFT treatment because of gastrointestinal toxicity. Conclusion: This schedule of UFT did not demonstrate a significant anti-tumor activity against metastatic pancreatic cancer.
Transcatheter Arterial Embolization with Zinostatin Stimalamer for Hepatocellular Carcinoma
Authors: Okusaka Takuji , Okada Shuichi , Ueno Hideki , Ikeda Masafumi , Iwata Ryoko , Furukawa Hiroyoshi , Takayasu Kenichi , Moriyama Noriyuki , Sato Tosiya , Sato Keiko
Published: April 2003

Zinostatin stimalamer (SMANCS) is a lipophilic intra-arterial chemotherapeutic agent for hepatocellular carcinoma (HCC). In our previous study, transcatheter arterial infusion chemotherapy using SMANCS for HCC showed a response rate of 20%. In an effort to obtain a superior anti-tumor effect against HCC, we conducted a phase II study of transcatheter arterial embolization (TAE) using SMANCS and gelatin sponge in 50 chemotherapy-naive patients with HCC. Four milligrams SMANCS plus 4 ml lipiodol emulsion was injected into the hepatic artery, followed by an injection of gelatin sponge. The responses were evaluated by computed tomography (CT) 1 month after treatment and thereafter every 3–4 months. One patient (2%) showed complete response and 15 patients (30%) had partial response resulting in an overall response rate of 32% (16/50; 95% confidence interval 19–45%). In 33 patients (66%), the disease remained stable, and 1 patient (2%) showed progressive disease. In 35 patients (70%), the rate of necrotic area to whole tumor was more than 50% according to the evaluation method using lipiodol accumulation in CT. The 1-, 3- and 5-year survival rates were 90, 55 and 19%, respectively. Grade 3 hematological toxicity was observed as thrombocytopenia in 2 patients (4%). Grade 3 and 4 non-hematological toxicity (liver dysfunction) occurred in 17 (34%) and 7 patients (14%), respectively. TAE using SMANCS, which was well tolerated, may be an effective treatment for advanced HCC.
mdm2-p53 Interaction: Lack of Correlation with the Response to 5-Fluorouracil in Advanced Colorectal Cancer
Authors: Paradiso A. , Ranieri G. , Simone G. , Silvestris N. , Costa A. , de Lena M. , Leone A. , Vallejo C. , Lacava J.
Published: April 2003

Objective: We investigated the relevance of mdm2 and p53 primary tumour expression to the clinical outcome of a consecutive series of advanced colorectal cancer patients treated with a 5-fluorouracil-based chemotherapy. Methods: The expression of p53 and mdm2 was analyzed by an immunohistochemical assay in 80 formalin-fixed paraffin embedded primary tumour samples and related to the clinical response to 5-fluorouracil therapy and to the prognosis of the patients. In a subgroup of 46 tumours, the apoptotic index (AI) as determined by the Tunel technique was also assessed. Results: Nuclear immunostaining of p53 and mdm2 was present in 42 and 30% of the cases, respectively. No correlation was demonstrated between p53 and mdm2 expression (rs = –0.01; p > 0.05). With regard to the clinical outcome, no statistical association was demonstrated among p53 and mdm2 expression, AI, probability of clinical response to treatment, time to progression, or overall survival. The subgroup of patients with a p53-negative/mdm2-positive tumour showed a worse response rate (15%); however, mdm2-positive/AI-negative tumours showed a 0% (0/7) probability of a clinical response as compared with 30% (9/30) of the remaining tumour patient subgroups; this also translated in a significantly worse overall survival probability (p = 0.01 by log rank). Conclusions: The analysis of mdm2 expression does not add significant clinical information in colorectal cancers with a different p53 status. Conversely, further analysis of AI seems to give data of a promising prognostic-predictive value.
ICAM-1 Expression on the Surface of T Lymphocytes in Patients with Uveitis: A Comparative Study between the Eye and Peripheral Blood
Authors: Marcos Martin Carmen , Santos Lacomba Manuel , Serrano Molina Carlos Ignacio , Ramirez Chamond Rafael , Collantes Estevez Eduardo
Published: April 2003

Purpose: We studied the surface expression of intracellular adhesion molecule 1 (ICAM-1) on peripheral and intra-ocular T lymphocytes in patients with active uveitis. Methods: Two-colour flow-cytometric analysis was performed on cells isolated from aqueous humour and peripheral blood of 23 patients with active uveitis and 16 control patients who were to undergo cataract extraction, in order to determine the percentage of cells expressing CD4, CD8 and ICAM-1 (CD54) molecules. Results: In the aqueous humour of patients with uveitis, we found an increase in the percentage of CD4+ and CD8+ lymphocytes, co-expressing the ICAM-1 molecule as compared to control patients (p < 0.0001). In peripheral blood, these uveitis patients exhibited a significant decrease in the percentage of CD4+ICAM-1+ (p = 0.0106) and CD8+ICAM-1+ (p = 0.0014) as compared to control subjects. The comparative study of cells from the aqueous humour and peripheral blood showed that the percentage of CD4+ICAM-1+ cells was higher in the aqueous humour (p < 0.0001). Comparison of the aqueous humour and peripheral blood for CD8+ICAM-1+ cells revealed no significant differences. In addition, we found a high negative correlation for the CD8+ICAM-1+ subset between the aqueous humour and peripheral blood. Conclusions: Our results suggest a greater local participation for CD4+ICAM-1+ cells but not for CD8+ICAM-1+ cells in the pathogenesis of uveitis.