bluuebunny/crossref_metadata_2025 huggingface dataset:
MASE1 and MASE2: Two Novel Integral Membrane Sensory Domains
Authors: Nikolskaya Anastasia N. , Mulkidjanian Armen Y. , Beech Iwona B. , Galperin Michael Y.
Published: April 2003

Escherichia coli proteins YegE and YaiC contain N-terminal integral membrane regions, followed by the putative diguanylate cyclase (GGDEF, DUF1) domains. The membrane domains of these proteins, named MASE1 (membrane-associated sensor) and MASE2, respectively, were found in other bacterial signaling proteins, such as histidine kinases (MASE1) and an adenylate cyclase (MASE2). Although the nature of the signals sensed by MASE1 and MASE2 is still unknown, MASE1-containing receptors appear to play important roles in bacteria, including iron and/or oxygen sensing by hemerythrine-containing proteins in the sulfate-reducing bacterium Desulfovibrio vulgaris.
O-Glycosylation of a Recombinant Carbohydrate-Binding Module Mutant Secreted by Pichia pastoris
Authors: Boraston Alisdair B. , Sandercock Linda E. , Warren R. Antony J. , Kilburn Douglas G.
Published: April 2003

Carbohydrate-binding modules (CBMs; previously called cellulose-binding domains) make excellent fusion partners for the immobilization or purification of polypeptides. However, their use in eukaryotic hosts has been limited by glycosylation, which interferes with the ability of the CBM to bind to cellulose. We have engineered the C-terminal carbohydrate-binding module from Cellulomonas fimi xylanase 10A such that it lacks N-glycosylation sites. This variant, called CBM2aNgly–, was produced and secreted by the methylotrophic yeast Pichia pastoris and found to be O-glycosylated. The O-linked glycans were composed entirely of mannose in a ratio of 1 mol of mannose to 4 mol of protein. The overall distribution of mannose on the O-glycosylated CBM mutant ranged from 1 to 9 mannose residues with the oligosaccharide sizes ranging from Man1 to Man4. MALDI-TOF (all matrix-assisted-laser-desorption time of flight) mass spectrometry (MS) was used to map the O-glycosylation to three regions of the polypeptide, each region having a maximum of 4 mannose residues attached to each. Glycans chemically released from CBM2aNgly– and analyzed by fluorophore-assisted carbohydrate electrophoresis were found to contain α-1,2-, α-1,3-, and α-1,6-linkages. Significantly, the O-glycosylation did not influence binding, making CBM2aNgly– a suitable fusion partner for polypeptides produced in P. pastoris and other eukaryotic hosts.
Massive Spontaneous Perirenal Hematoma and Accelerated Hypertension in a Patient with Polyarteritis nodosa
Authors: Minardi D. , Dessì-Fulgheri P. , Sarzani R. , Onesta M. , Muçaj A. , Branchi A. , Giangiacomi M. , Mantovani P. , Muzzonigro G.
Published: April 2003

A 43-year-old Albanian man is presented who underwent nephrectomy for a huge right spontaneous perirenal hematoma. The diagnosis of polyarteritis nodosa as the etiology of the hematoma has been made only by histological examination, because of the quick and unforeseeable onset of this complication and the nonspecificity of symptoms. We hypothesize a relationship between reactivation of polyarteritis nodosa and treatment with rifampicin and isoniazid.
Regional Cerebral Blood Flow Patterns and Response to Donepezil Treatment in Patients with Alzheimer’s Disease
Authors: Hanyu Haruo , Shimizu Takeshi , Tanaka Yuriko , Takasaki Masaru , Koizumi Kiyoshi , Abe Kimihiko
Published: April 2003

We attempted to identify the characteristic patterns in regional cerebral blood flow (rCBF) of patients with Alzheimer’s disease who responded to donepezil therapy. Sixty-one patients treated with donepezil were divided into two groups (responders and nonresponders) on the basis of changes in their Mini Mental State Examination (MMSE) scores from baseline to study endpoint. We analyzed single-photon emission computed tomography data (SPECT) at baseline using three-dimensional stereotactic surface projections and compared differences in rCBF between the two groups. Statistical maps showed a significantly lower rCBF of the lateral and medial frontal lobes in the nonresponders than in the responders. There was a significant inverse correlation between the relative rCBF reduction in the frontal lobe and the MMSE change. These results suggest that frontal function, as assessed by SPECT, affects the patient’s response to treatment with donepezil. Measuring rCBF may aid in the selection of possible treatment responders.
C.G. Jung – ein unerkannter Systemtheoretiker?
Authors: Grosskreutz Albrecht
Published: April 2003

Entgegen der Behauptung Toblers, Jung habe der systemtheoretische Ansatz bei der Formulierung seiner Theorie gefehlt, kann belegt werden, dass Jung systemorientiert dachte, seine Erkenntnisse jedoch nicht in eine Systemtheorie, sondern in einen geisteswissenschaftlichen Kontext einzubringen suchte. Kritisiert wird Toblers Versuch, allein mit Hilfe der Systemtheorie erkenntnisleitend vorzugehen, da dies nicht nur zu einer inadäquaten Annäherung an die Analytische Psychologie führt, sondern auch die Lebenswirklichkeit des historisch-empirischen Menschen im 21. Jahrhundert, die durch die Aufgabe der Bewältigung einer Vielzahl von Dissoziationen gekennzeichnet ist, übergeht. Beispielhaft wird dies am systemischen Begriff der Autopoiese gezeigt, die eine Auffassung der Wirklichkeit unter dem Aspekt der Einheitlichkeit und Harmonie suggeriert, während erkenntnisleitend und lebenspraktisch von einem Konzept der Diskontinuität auszugehen ist.
Primary Splenic Hydatidosis
Authors: Durgun Vedat , Kapan Selin , Kapan Metin , Karabiçak Ilhan , Aydogan Fatih , Goksoy Ertugrul
Published: April 2003

Background: A hydatid cyst is still a serious health problem in endemic areas. Invasion of the spleen is rare in hydatid disease. In the medical literature, reports about isolated splenic hydatidoses are quite rare. Also there is not a consensus about the way of treatment. Methods: Between 1978 and 2000, in our clinic approximately 900 patients were operated on for an abdominal hydatidosis; 850 of these patients were treated for a hepatic hydatidosis and 14 patients for an isolated splenic hydatidosis. Six (42.8%) of the latter patients were male and 8 (57.1%) were female. The mean age of the patients was 47.14 ± 4.9 (range 17–72) years. Ten patients (71.4%) presented with a painful mass in the left upper quadrant of the abdomen, and the other 4 patients (28.5%) were asymptomatic and were diagnosed incidentally. Results: All of the patients underwent elective splenectomy. There was no mortality, but complications occurred in 4 (28.5%) patients. The period of hospitalization ranged from 7 to 17 days with a mean of 9.8 days. 1 patient died from an acute myocardial infarction during the 2nd postoperative year, and 1 patient died as a consequence of a traffic accident during the 5th postoperative year. Nine patients, after follow-up periods of between 2 and 14 years, are living free from disease. No recurrence occurred in any of them. In the remaining 3 patients, long-term follow-up could not be maintained. Conclusions: A hydatid cyst must be included in the differential diagnosis of cystic lesions of the spleen. A splenic hydatid cyst should be treated surgically due to the high risk of a rupture, and the ideal procedure in adulthood is standard splenectomy.
Somites, Spinal Ganglia, and Centra
Authors: O'Rahilly R. , Müller F.
Published: April 2003

Serial sections of 99 human embryos from Carnegie stages 8–23 were investigated and 38 graphic reconstructions were evaluated. At stage 9 somite 1 is of appreciable size and is separated from the otic disc, as also in the next several stages by rhombomeres and pharyngeal arches 3 and 4, thereby differing from the chick. At stage 10 somite 1 begins to differentiate into sclerotome and dermatomyotome. At stage 11 spinal neural crest begins to develop. At stage 12 parts of somites 1–4 are being transformed into the hypoglossal cell cord. It is stressed that the numbers of somites present at stages 9–12 are part of the definition of those stages. At stage 13 dense and loose zones begin to be detectable rostrally in the sclerotomes and also, although out of phase, in the perinotochord. Spinal ganglia begin to develop in phase with the somites. At stages 14–16 the maximum number of somites observed was 38–39 rather than 42–44, as usually given. Moreover, they did not extend to the tapered end of the trunk, which is not a (vertebrated) ‘tail’. At stages 17–23 the maximum number of centra was 38–39, including coccygeal vertebrae 4–5. Although most of the somites appear during primary development, all of the spinal ganglia develop during secondary development (stages 13–18). The number of ganglia was at a maximum of 35 at stage 18, but was reduced to 32 already by stage 23. Important points confirmed in this study are that the number of occipital somites in the human is four, and that the level of final closure of the caudal neuropore is future somite 31, which represents approximately future sacral vertebra 2. The interpretation of relevant neural tube defects is discussed in the light of the findings. The ascensus of the conus medullaris during the fetal period is well established, but a concomitant ascent of the situs neuroporicus is proposed here, and has implications for defects that involve tethering of the spinal cord. The main results are integrated in comprehensive graphic representations of the levels and the interrelationships of (a) somites and centra, and (b) somites, neural crest, and spinal ganglia. These may aid in the elucidation of some frequently occurring anomalous conditions.
Long-Term Administration of Intravaginal Dehydroepiandrosterone on Regression of Low-Grade Cervical Dysplasia – A Pilot Study
Authors: Suh-Burgmann Elizabeth , Sivret Julia , Duska Linda R. , Del Carmen Marcella , Seiden Michael V.
Published: April 2003

Although many dysplastic cervical lesions regress spontaneously, treatment is common due to concern for progression. Lesions persist or progress in women whose immune systems are unable to clear infection by human papillomavirus (HPV). Dehydroepiandrosterone (DHEA) is an adrenal steroid that has both immune modulatory and tumor inhibitory activity. A pilot study was conducted to examine the feasibility, safety and potential efficacy of intravaginal DHEA in women with low-grade cervical dysplasia. Twelve women with low-grade dysplasia, confirmed by colposcopic exam, were given 150 mg of intravaginal micronized DHEA daily for up to 6 months. Follow-up evaluations of the cervix were done at 3 and 6 months of use. DHEA, DHEA-S, androstenedione and testosterone levels were also measured. By the end of the study period, 10 of the 12 women (83%) had no evidence of dysplasia; the remaining 2 had normal colposcopic exams but cytology showing atypical cells of undetermined significance. There were no serious side effects. Androstenedione levels were elevated at 3 months, whereas testosterone levels were unchanged over the course of treatment. The results suggest that intravaginal DHEA is safe and well tolerated and may promote regression of low-grade cervical lesions. Further study is needed to establish efficacy.
The Nature and Origin of Intraocular Metallic Foreign Bodies Appearing after Phacoemulsification
Authors: Köse Süheyla , Menteş Jale , Üretmen Önder , Topçuoğlu Nejat , Köktürk Uğur , Yilmaz Hatice
Published: April 2003

In order to evaluate the nature and origin of metallic foreign bodies embedded in the iris after uneventful phacoemulsification, we aimed to produce metallic fragments by applying ultrasound power with the same phaco machine and handpiece in an experimental model in vitro. In a glass bottle, we used linear phaco power of 100% continuously for 5 min first with a new phaco tip and then with a used tip. Afterwards, the fluid in the bottle was filtered through a Millipore filter. The remains on the filter were studied by scanning electron microscopy (SEM) and microprobe. Small particles remained on the filter which was used for the new tip. Microprobe microanalysis showed that these particles were mainly titanium, the same as the phaco tip. SEM of both the new and the used tips showed small fragments on the exterior surface and lumen. Intraocular metallic foreign bodies after phacoemulsification are likely to be shaken loose from the phaco tip. Although it is mostly agreed that these fragments are well tolerated, their overall effect remains to be evaluated in the long term.
Central Hearing Loss with a Bilateral Inferior Colliculus Lesion
Authors: Hoistad Dick L. , Hain Timothy C.
Published: April 2003

A case is presented of a 43-year-old man with bilateral lesions of the inferior colliculus (IC) due to central nervous system lymphoma. Our patient was found to have relatively normal pure-tone averages with severely reduced bilateral word recognition scores. This case illustrates that the presentation of a partial bilateral IC lesion may be of severely reduced word recognition accompanied by nearly normal pure-tone hearing.
Determination of IgG Subclasses and Avidity of Antithyroid Peroxidase Antibodies in Patients with Subclinical Hypothyroidism – A Comparison with Patients with Overt Hypothyroidism
Authors: Silva L.M. , Chavez J. , Canalli M.H.B. , Zanetti C.R.
Published: April 2003

Objective: To evaluate the immunoglobulin G subclasses of anti-TPO and antibody avidity in patients with subclinical hypothyroidism (sH), overt hypothyroidism (H) and a control group (C). Methods: According to the TSH, fT4 and anti-TPO antibody levels, appraised by immunometric assays, 95 female patients were divided into three groups (sH, H and C). IgG subclass levels and avidity were measured by a homemade ELISA. Results were analyzed by nonparametric tests and Spearman’s rank correlation. Results: The predominant IgG subclasses detected in both case groups were IgG1 and IgG4 with a significantly higher level of IgG4 in the sH group. Consequently, the IgG1/IgG4 ratio was significantly lower in sH patients. Conclusion: The higher levels of IgG4 anti-TPO reduced significantly the IgG1/IgG4 ratio in sH patients. These results permit to envisage that increasing this ratio could be useful as a positive predictive factor for the development of overt disease in such patients.
Statistical Parametric Mapping of Brain SPECT Perfusion Abnormalities in Patients with Alzheimer’s Disease
Authors: Lee Yi-Chung , Liu Ren-Shyan , Liao Yi-Chu , Sun Chen-Ming , Wang Po-Shan , Wang Pei-Ning , Liu Hsiu-Chih
Published: April 2003

Brain perfusion in 20 patients with mild Alzheimer’s disease (AD), 20 patients with moderate AD and 20 control subjects (matched for age, gender and education) were assessed by single photon emission computed tomography (SPECT) using technetium-99m hexamethylpropylene amine oxime (99mTc-HMPAO). SPECT images were transformed to a standard size and shape for group comparisons by the voxel-based t test of the statistical parametric mapping techniques. Cerebral hypoperfusion in the left lower parietal area was found in mild AD patients. In moderate AD patients, significant cerebral hypoperfusion was located in bilateral posterior parietotemporal cortices, contiguous anterior occipital lobes, posterior cingulate gyri and, to a lesser extent, in frontal areas.
Wheat Bran and Oat Bran Effectively Reduce Oxidative Stress Induced by High-Fat Diets in Pigs
Authors: Rezar V. , Pajk T. , Marinšek Logar R. , Ješe Janežič V. , Salobir K. , Orešnik A. , Salobir J.
Published: April 2003

Aim: To evaluate the influence of wheat bran and oat bran on the oxidative stress induced by a high proportion of fat in the diet. Methods: Forty-eight growing pigs were penned individually and after an adaptation period divided into four groups. All groups received isocaloric daily rations composed of basal diet which was then supplemented with: starch (controls; CONT), linseed oil (OIL), linseed oil and wheat bran, or linseed oil and oat bran. The experimental period lasted 14-days. The oxidative stress was evaluated by measuring the malondialdehyde (MDA) concentration in blood plasma, the 48-hour urinary MDA excretion, and the degree of leukocyte nuclear DNA damage. Results: In comparison with the CONT group, a significant increase in the MDA concentration in blood plasma and in the MDA excretion in urine was found in the OIL group. The degree of DNA damage in the OIL group was also significantly higher. In comparison with the OIL group, the wheat bran and oat bran supplementation significantly reduced the 24-hour MDA excretion in urine and reduced the degree of DNA damage in leukocytes to the level of the CONT group. Conclusion: The results of the experiments confirmed that a high wheat bran and oat bran intake effectively reduces oxidative stress induced by a high-fat diet.
Expression of COX-2 in Stomach Cancers and Its Relation to Their Biological Features
Authors: Han Shao-Liang , Tang Huai-Jing , Hua Ya-Wei , Ji She-Qing , Lin Dong-Xin
Published: April 2003

Background/Aim: Cyclooxygenase-2 (COX-2) is one of the key isoenzymes in the production of prostaglandins, and is believed to be involved in carcinogenesis. This study was conducted to examine the role of COX-2 in the development and biological behavior of stomach cancer. Methods: Expression of COX-2 at the mRNA and protein levels was analyzed using RT-PCR and immunoblotting assay in 50 cancerous and corresponding non-cancerous tissue specimens. Also, COX-2 expression was detected by an immunohistochemical method in 55 paraffin-embedded gastric adenocarcinoma tissues. Results: Of the 50 carcinoma tissue samples analyzed, 38 (76.0%) had overexpression of COX-2 as compared to the paired non-cancerous specimens. The overexpression of COX-2 (91.7%) was more prevalent in tumors with sizes of >5 cm than those (61.5%) of ≤5 cm (p < 0.05). COX-2 overexpression was higher in tumors with >6 metastatic nodes compared to those with ≤6 metastatic nodes (100.0 vs. 61.3%, p < 0.01). The rate of COX-2 overexpression in non-signet ring cell carcinoma was significantly higher compared with signet ring cell carcinoma (84.1 vs. 16.7%, p < 0.05). The rate of COX-2 overexpression (86.8 or 90.9%) in patients with a cancer invasion depth of pT3 + pT4 or with stage III + IV was significantly higher than that (41.7 or 47.1%) in patients with pT1 + pT2 or with stage I + II (both p < 0.01). Furthermore, consistent with the results at the protein and mRNA levels, immunohistochemical stain indicated that COX-2 protein was overexpressed in 39 of 55 (70.9%) carcinomas, and the 5-year survival rate (25.6%) for patients with COX-2 overexpression seemed better than that (50.0%) of patients without overexpression, though the difference was not significant (p < 0.05). Conclusion: Our results indicate that COX-2 may play a role in the development of gastric cancer and its overexpression is associated with lymphatic metastasis, tumor invasion and differentiation of gastric carcinoma.
Multiple Brain Infarcts: Clinical and Neuroimaging Patterns Using Diffusion-Weighted Magnetic Resonance
Authors: Altieri M. , Metz R.J. , Müller C. , Maeder P. , Meuli R. , Bogousslavsky J.
Published: April 2003

The capability of diffusion-weighted (DW) magnetic resonance imaging (MRI) to identify very early ischemic brain injury better than conventional MRI is well known. This technique, which successfully discriminates acute from old infarcts, is particularly useful in patients with multiple brain infarcts (MBI). Among 142 patients with acute stroke consecutively admitted to our primary care center, we selected 43 patients with two or more brain infarcts on conventional MRI. All patients presented with clinical deficits consistent with acute cerebral ischemia and underwent conventional spin echo for T1 (T1-WI) and T2-weighted images (T2-WI), T1-W gadolinium-enhanced images, and echo-planar technique for DW MRI sequences. Patients underwent DW MRI examinations within 15 days of stroke onset (mean ± SD: 3 ± 3 days). In all but 1 case, the infarcts detected on DW MRI were also visible on T2-WI. The different signal pattern on DW MRI, compared with T2-WI, facilitated the detection of acute infarcts in all patients. T1-WI with gadolinium enhancement was only helpful in 5 (11.6%) patients. DW MRI enabled precise clinicotopographic correlations in 79% of our patients and provided additional clinically relevant findings in 72% of the patients. Based on the neuroradiological findings, patients were divided into three clinicotopographic types of MBI as follows: 13 patients (30.2%) presented with multiple acute infarcts, 24 patients (55.8%) with a single acute infarct and multiple old infarcts, and 6 patients (13.9%) with multiple acute and old infarcts. In conclusion, DW MRI can easily be added to conventional MRI in order to be able to distinguish acute from old infarcts, and to identify acute multiple lesions. Therefore, a better correlation between clinical symptoms and the site of lesions can be obtained, considerably improving patient care.
Fibre Composition of Human Intrinsic Tongue Muscles
Authors: Stål P. , Marklund S. , Thornell L.-E. , De Paul R. , Eriksson P.-O.
Published: April 2003

The muscle fibre composition of three human intrinsic tongue muscles, the longitudinalis, verticalis and transversus, was investigated in four anterior to posterior regions of the tongue using morphological and enzyme- and immunohistochemical techniques. All three muscles typically contained type I, IIA and IM/IIC fibres. Type I fibres expressed slow myosin heavy chain (MyHC), type II fibres fast MyHC, mainly fast A MyHC, whereas type IM/IIC coexpressed slow and fast MyHCs. Type II fibres were in the majority (60%), but regional differences in proportion and diameter of fibre types were obvious. The anterior region of the tongue contained a predominance of relatively small type II fibres (71%), in contrast to the posterior region which instead showed a majority of larger type I and type IM/IIC fibres (66%). In general, the fibre diameter was larger in the posterior region. This muscle fibre composition of the tongue differs from those of limb, orofacial and masticatory muscles, probably reflecting genotypic as well as phenotypic functional specialization in oral function. The predominance of type II fibres and the regional differences in fibre composition, together with intricate muscle structure, suggest generally fast and flexible actions in positioning and shaping the tongue, during vital tasks such as mastication, swallowing, respiration and speech.
Immunolocalization of Fibronectin and Collagen Types I and III in Human Fetal Parotid and Submandibular Glands
Authors: Kumagai Mitsutake , Sato Iwao
Published: April 2003

We examined the distribution of fibronectin and collagen types I and III in human fetuses under a confocal laser scanning microscope using immunohistochemical staining (at 16, 20, 24, 28, 32 weeks’ gestation). The collagen types I and III form the collagenous matrix components in the connective tissue of the parotid and submandibular glands. These extracellular matrix components were detected at various stages around the terminal portion and in the capsule-like connective tissue of the parotid and submandibular glands. However, the extracellular matrix components in the connective tissue around the terminal portion had a stronger reaction than those in the capsule-like connective tissue found on the fringe of the terminal portion. The collagen type I of the parotid gland at 16 weeks’ gestation had a weaker reaction than that of the submandibular gland. When results of the reaction at other stages and other extracellular matrix components of the two salivary glands were compared, collagen type I appeared as early as at 16 weeks’ gestation in either gland. Since collagen type I serves as the core connective tissue, these observations suggest that the formation of connective tissue around the parotid gland occurs before or after 16 weeks’ gestation lagging behind that of the submandibular gland.
A Confocal Microscopic Study of Gonadotropin-Releasing Hormone (GnRH) Neuron Inputs to Dopaminergic Neurons Containing Estrogen Receptor α in the Arcuate Nucleus of GnRH-Green Fluorescent Protein Transgenic Mice
Authors: Mitchell Valerie , Loyens Anne , Spergel Daniel J. , Flactif Martial , Poulain Pierre , Tramu Gerard , Beauvillain Jean-Claude
Published: April 2003

The purpose of the present study was to determine whether the septo-preoptico-tuberoinfundibular gonadotropin-releasing hormone (GnRH) pathway comes in close juxtaposition with tyrosine hydroxylase immunoreactive (TH-IR) neurons in the arcuate nucleus of female mice. Immunohistochemical staining with a TH monoclonal antibody coupled with confocal microscopy was employed on vibratome-cut brain sections of female GnRH-green fluorescent protein (GFP) transgenic mice to evaluate possible appositions between GnRH and tuberoinfundibular dopaminergic (TIDA) neurons. TH-IR neurons of the arcuate nucleus received GnRH neuronal appositions in adult female mice at proestrus and estrus stages. In contrast, no GnRH appositions were observed in adult females at diestrus. Subsequently, double immunohistochemical staining for TH and estrogen receptor-α (ERα) was performed to examine the role of estradiol on this relationship. We found that most TH-IR neurons contacted by GnRH fibers were immunoreactive for ERα. Our observations suggest that GnRH neurons communicate directly with TIDA neurons in the adult female. Furthermore, ERα activation in TIDA neurons may be involved in the formation of connections between GnRH neurons and TIDA neurons.
Antiproliferative Activity of the Extract of Gleditsia sinensis Fruit on Human Solid Tumour Cell Lines
Authors: Chow L.M.C. , Tang J.C.O. , Teo I.T.N. , Chui C.H. , Lau F.Y. , Leung T.W.T. , Cheng G. , Wong R.S.M. , Wong I.L.K. , Tsang K.M.S. , Tan W.Q. , Zhao Y.Z. , Lai K.B. , Lam W.H. , Guo D.A. , Chan A.S.C.
Published: April 2003

Background: The fruit extract of Gleditsia sinensis Lam. (GSE) is a traditional herbal medicine that is saponin-rich. However, its activity on solid tumour cell lines has never been demonstrated. Methods: The activity of GSE was demonstrated in four cancer cell lines (breast cancer MCF-7, MDA-MB231, hepatoblastoma HepG2 and oesophageal squamous carcinoma cell line SLMT-1) using MTT assay, anchorage-independent clonogenicity assay, DNA laddering and in situ cell death detection. Results: The mean MTT50 (the mean concentration of GSE to reduce MTT activity by 50%) ranged from 16 to 20 µg/ml of GSE. An anchorage-independent clonogenicity assay showed that all of the four solid tumour cell lines gradually lost their regeneration potential after treatment with GSE, DNA fragmentation and TUNEL analysis demonstrated that the action of GSE is both dose- and time course-dependent. Conclusions: Our results suggest that GSE has a cytotoxic activity and can induce apoptosis in human solid tumour cell lines.
Establishment of an R-banded rabbit karyotype nomenclature by FISH localization of 23 chromosome-specific genes on both G- and R-banded chromosomes
Authors: Hayes H. , Rogel-Gaillard C. , Zijlstra C. , de Haan N.A. , Urien C. , Bourgeaux N. , Bertaud M. , Bosma A.A.
Published: April 2003

Direct detection of fluorescent in situ hybridization signals on R-banded chromosomes stained with propidium iodide is a rapid and efficient method for constructing cytogenetic maps for species with R-banded standard karyotypes. In this paper, our aim is to establish an R-banded rabbit karyotype nomenclature that is in total agreement with the 1981 G-banded standard nomenclature. For this purpose, we have produced new GTG- and RBG-banded mid-metaphase karyotypes and an updated version of ideograms of R-banded rabbit chromosomes. In addition, to confirm correlations between G- and R-banded chromosomes, we have defined a set of 23 rabbit BAC clones, each containing a specific gene, one marker gene per rabbit chromosome, and we have localized precisely each BAC clone by FISH on both G- and R-banded chromosomes.
Molecular characterization of the porcine gene CAPNS1 encoding the small subunit 1 of calpain on SSC6q1.1→q1.2
Authors: Drögemüller C. , Leeb T.
Published: April 2003

The CAPNS1 gene encodes the small subunit of the calpain proteases. Cloning and characterization of the porcine CAPNS1 gene revealed a highly conserved organization with respect to other mammalian CAPNS1 orthologs. The porcine gene consists of 11 exons spanning approximately 7.3 kb and codes for a peptide of 266 amino acids. Analysis of the naturally occurring variation of the CAPNS1 gene in different pig breeds revealed six SNPs and four small insertion/deletion polymorphisms. The CAPNS1 gene is located immediately adjacent to the COX7A1 gene on SSC6q1.1→q1.2.
THERMAL STABILITY OF ELECTRODEPOSITED NANOCRYSTALLINE NICKEL
Authors: Abraham M. , Holdway P. , Thuvander M. , Cerezo A. , Smith G. D. W.
Published: April 2003

The effect of annealing electrodeposited nanocrystalline nickel at 250°C was studied by microhardness testing, XRD, TEM and three-dimensional atom probe (3DAP) analysis. TEM micrographs revealed that the type of growth is abnormal, i.e. a composite structure is formed with some large (>1 μm) grains embedded in a matrix of nanometre sized grains. 3DAP studies showed no grain boundary segregation in as deposited materials and only low levels in material annealed for 1 h. Upon annealing, a 10% increase in hardness was observed within the first five minutes, after which the hardness was maintained for long times, despite the high volume fraction of large grains. Only after extensive annealing, when the majority of the material was made up of large grains, did the hardness decrease rapidly. The results demonstrate that the hardness is not determined by a law of mixtures but by percolation of the large grains.
Tungsten Laser Alloying of A356 Al Alloy - Tribological Performance and Characterisation
Authors: Staia M. H. , Sanchez C. , Cruz M. R. , Roman A. , Lesage J. , Mesmacque G. , Dahotre N. B.
Published: April 2003

In the present work, sand cast Al alloy A356 was laser alloyed with tungsten using a two stage technique of laser alloying. A continuous wave, fibre optically delivered, Nd-YAG laser was employed for alloying the tungsten powder, and the coatings were produced by varying beam interaction with the substrate by altering the traverse laser velocity. Characterisation of the laser treated alloy was performed by various techniques. Optical (OM) and scanning electron microscopy (SEM) were used to examine the morphology and microstructure of the specimen surfaces and cross-sections. Surface layers alloyed with tungsten contained relatively large amounts of intermetallic compounds such as Ti5Si4, Mg2Si, Al5 W, Al12W, and Al3·21Si0·47 identified by X-ray diffraction. Friction and wear tests were carried out on all specimens by using a ball on disc tribometer under dry conditions. The static partners were balls of 6 mm diameter made of WC + 6%Co. It was determined that the laser treated surfaces presented an increase in wear resistance between 40% and nearly 110% when compared to the wear resistance of the untreated Al alloy, and the improvement was proportional with the laser beam- substrate interaction time.
Corrosion wear response of S phase coated 316L
Authors: Dearnley P. A.
Published: April 2003

S phase is a metastable nitrogen supersaturated fcc phase which, when applied as a coating using reactive magnetron sputtering, can significantly improve the corrosion wear resistance of austenitic stainless steels. Coatings with nitrogen ranges between 0.3 and 44 at.-%N were analysed and tested for hardness, modulus and fracture resistance. Duplex bcc + fcc coatings decreased in hardness with increasing N content, whereas the hardness of monophase S phase coatings increased, reaching a maximum of ∼20 GPa at 28-32 at.-%N. Three types of corrosion wear process affect S phase coated 316L: first, removal of the passive film during sliding contact, second, galvanic attack of the substrate and third, galvanic attack of the counterface material. These three types are discussed and solutions suggested.
Properties and Performance Crtialn of Multilayer Hard Coatings Deposited Using Magnetron Sputter Ion Plating
Authors: Yang S. , Teer D. G.
Published: April 2003

A new CrTiAlN coating has been produced using closed field magnetron sputter ion plating and individual Ti, Al and Cr metal targets. M42 high speed steel coupons, 30 × 30 × 3.0 mm stainless steel (316) and 6.35 mm diameter (¼in) high speed steel twist drills were used as substrates. The coating was structured as follows: a Cr rich metal adhesive layer; a monolithic CrN nitride columnar layer with increasing content of Ti and Al; and finally, the nanoscale CrTiAlN multilayer. The composition ratio of Cr, Ti and Al in the multilayer was optimised and the nitrogen flow was controlled using an optical emission monitor with feedback control to produce stoichiometric CrTiAlN (metal/N ∼50:50). Some of the as coated samples were heat treated in air at 600°C and 900°C, and then the coating compositions and structures were studied using XRD, GDOES, and TEM with special reference to the effects of the heat treatment. The hardness, adhesion and wear properties of the coating were studied and the properties of the as coated and the heat treated samples were compared. It has been shown that the coatings have many desirable properties (hardness, toughness, wear resistance and thermal stability) at high level. The main part of the coating was formed as a nanoscale multilayer and this coating was applied on to twist tool steel drills giving excellent drilling performance.
Synergy between tumor immunotherapy and antiangiogenic therapy
Authors: Nair Smita , Boczkowski David , Moeller Benjamin , Dewhirst Mark , Vieweg Johannes , Gilboa Eli
Published: April 2003

This study tested the hypothesis that combination of antiangiogenic therapy and tumor immunotherapy of cancer is synergistic. To inhibit angiogenesis, mice were immunized with dendritic cells (DCs) transfected with mRNA that encode products that are preferentially expressed during neoangiogenesis: vascular endothelial growth factor receptor-2 (VEGFR-2) and Tie2 expressed in proliferating endothelial cells, and vascular endothelial growth factor (VEGF) expressed in the angiogenic stroma as well as the tumor cells used in this study. Immunization of mice against VEGF or VEGFR-2 stimulated cytotoxic T lymphocyte (CTL) responses and led to partial inhibition of angiogenesis. Antiangiogenic immunity was not associated with morbidity or mortality except for a transient impact on fertility seen in mice immunized against VEGFR-2, but not VEGF. Tumor growth was significantly inhibited in mice immunized against VEGF, VEGFR-2, and Tie2, either before tumor challenge or in the setting of pre-existing disease in murine B16/F10.9 melanoma and MBT-2 bladder tumor models. Coimmunization of mice against VEGFR-2 or Tie2 and total tumor RNA exhibited a synergistic antitumor effect. Synergism was also observed when mice were coimmunized with various combinations of defined tumor-expressed antigens, telomerase reverse transcriptase (TERT) or TRP-2, and VEGF or VEGFR-2. This study shows that coimmunizing mice against angiogenesis-associated and tumor-expressed antigens can deliver 2 compatible and synergistic cancer treatment modalities via a common treatment, namely immunization.
Multidrug resistance protein attenuates gemtuzumab ozogamicin–induced cytotoxicity in acute myeloid leukemia cells
Authors: Walter Roland B. , Raden Brian W. , Hong Tom C. , Flowers David A. , Bernstein Irwin D. , Linenberger Michael L.
Published: April 2003

Gemtuzumab ozogamicin (GO) is a novel immunoconjugate therapy for acute myeloid leukemia (AML). P-glycoprotein (Pgp) confers resistance to GO and is associated with a worse clinical response. To address whether multidrug resistance protein (MRP) affects GO susceptibility, we characterized Pgp, MRP1, and MRP2 expression in CD33+ cell lines and CD33+ AML samples and analyzed the effect of the Pgp inhibitor cyclosporine (CSA) and the MRP inhibitor MK-571 on GO-induced cytotoxicity. MRP1, but not MRP2, expression correlated with MRP activity. MK-571 enhanced GO-induced cytotoxicity in Pgpnegative/MRP-positive NB4 and HL-60 cells. CSA, but not MK-571 alone, restored GO susceptibility in Pgp-positive/MRP-positive TF1 cells; however, MK-571 enhanced cytotoxicity in the presence of CSA. All patient samples exhibited MRP activity, and 17 of 23 exhibited Pgp activity. CSA increased GO-induced cytotoxicity in 12 Pgp-positive samples, whereas MK-571 alone was effective in only one sample with minimal Pgp activity. In 3 Pgp-positive/MRP-positive samples, MK-571 enhanced GO-induced cytotoxicity in the presence of CSA. Thus, MRP1 may attenuate susceptibility to GO. This effect was comparatively less than that for Pgp and required the inhibition of Pgp for detection in cells that coexpressed both transporters. Because MK-571 and CSA failed to affect cytotoxicity in a portion of Pgp-positive/MRP-positive AML samples, additional resistance mechanisms are likely important.
Tissue engineering and the trauma surgeon
Authors: Garner JP
Published: April 2003

The demands for repair and renewal of worn out or injured human tissue grow year on year it is a demand that cannot be met from live human donors. A partial solution may be found from cadaveric or trans-species transplantation of tissue, but these approaches are steeped in the problems of disease transfer and ethical dilemma. Tissue engineering is a new technology that seeks to meet these increasing demands by utilizing novel cell culture methods in vitro to provide tissue replacements in vivo. This article reviews the current state of tissue engineering and its potential for use in the realms of trauma surgery.
Synovial Sarcoma of Childhood and Adolescence: A Multicenter, Multivariate Analysis of Outcome
Authors: Okcu Mehmet Fatih , Munsell Mark , Treuner Joern , Mattke Adrian , Pappo Alberto , Cain Alvida , Ferrari Andrea , Casanova Michela , Ozkan Alp , Raney Beverly
Published: April 2003

Purpose: To identify prognostic factors related to outcome in 219 children and adolescents with synovial sarcoma.

Patients and Methods: We combined the experiences of the four following research groups: Cooperative Weichteilsarkomastudie Group, Germany (n = 95); St. Jude Children’s Research Hospital, Memphis, TN (n = 49); Istituto Nazionale dei Tumori, Milan, Italy (n = 33); and The University of Texas M.D. Anderson Cancer Center, Houston, TX (n = 42). Kaplan-Meier and Cox proportional hazard analyses were performed.

Results: The median age at diagnosis was 13 years (range, 1 to 20 years), and the median follow-up was 6.6 years (range, 0.5 to 30.7 years). The estimated 5-year overall survival and event-free survival rates for the entire group were 80% ± 3% (SE) and 72% ± 3%, respectively. A previously unreported interaction between tumor size and invasiveness was observed that statistically significantly related to outcome. In multivarible analysis, patients with T1B and T2B disease (hazard ratio [HR] = 5.6, 95% confidence interval (CI), 1.9 to 16.2; and HR = 5.9, 95% CI, 2.1 to 16.4, respectively) or Intergroup Rhabdomyosarcoma Study (IRS) Clinical Group III and IV disease (HR = 2.7, 95% CI, 1.2 to 6.5; and HR = 14.1, 95% CI, 4.3 to 31.3, respectively) had poor overall survival. Treatment with radiotherapy was related to improved overall survival (HR = 0.4; 95% CI, 0.2 to 0.7). In IRS Group III patients, objective response to chemotherapy (18 of 30, 60%) correlated with improved survival.

Conclusion: Clinical group, tumor size, and invasiveness are important prognostic factors. Multicenter randomized clinical trials are needed to determine both the effect of chemotherapy on survival and the necessity of local radiotherapy in patients with completely resected tumors.
Provider Delay Among Patients With Breast Cancer in Germany: A Population-Based Study
Authors: Arndt Volker , Stürmer Til , Stegmaier Christa , Ziegler Hartwig , Becker Annelie , Brenner Hermann
Published: April 2003

Purpose: Delaying the diagnosis and initiation of treatment of cancer is likely to result in tumor progression and a worse prognosis. We examined sources and consequences of provider delay among female breast cancer patients in a population-based study in Germany.

Patients and Methods: Three hundred eighty women, who were ages 18 to 80 years and who had invasive breast cancer, were interviewed with respect to the diagnostic process. Provider delay was defined as time from first presentation to a health care provider until initiation of cancer treatment.

Results: Median provider delay was 15 days and did not substantially differ by the specialty of first consulted physician. Delays in the diagnostic work-up were mainly because of erroneous initial suspicion of a benign breast disease or because of time constraints by patients or physicians. Provider delay over 3 months was found in 11% of all breast cancer cases and was associated with patient characteristics such as higher education (odds ratio [OR] = 2.6; 95% confidence interval [CI], 1.3 to 5.4), full-time employment (OR = 2.5; 95% CI, 1.1 to 5.5), family history of breast cancer (OR = 2.8; 95% CI, 1.2 to 6.2), and presenting with a non–breast symptom (OR = 4.3; 95% CI, 1.7 to 10.9). The association between duration of diagnostic work-up and stage at diagnosis was U shaped, with the highest proportions of metastasized breast cancer tumors among women with very short (< 7 days) or very long (> 3 months) duration.

Conclusion: Diagnostic work-up is within reasonably short time limits among most patients with breast cancer in Germany. Although the association between delay and tumor stage seems to be complex, any delay in diagnostic work-up should be kept to a minimum.
Adjuvant Immunization of HLA-A2–Positive Melanoma Patients With a Modified gp100 Peptide Induces Peptide-Specific CD8+T-Cell Responses
Authors: Smith John W. , Walker Edwin B. , Fox Bernard A. , Haley Daniel , Wisner Ketura P. , Doran Teri , Fisher Brenda , Justice Lisa , Wood William , Vetto John , Maecker Holden , Dols Annemiek , Meijer Sybren , Hu Hong-Ming , Romero Pedro , Alvord W. Gregory , Urba Walter J.
Published: April 2003

Purpose: To measure the CD8+T-cell response to a melanoma peptide vaccine and to compare an every-2-weeks with an every-3-weeks vaccination schedule.

Patients and Methods: Thirty HLA-A2–positive patients with resected stage I to III melanoma were randomly assigned to receive vaccinations every 2 weeks (13 vaccines) or every 3 weeks (nine vaccines) for 6 months. The synthetic, modified gp100 peptide, g209–2M, and a control peptide, HPV16 E7, were mixed in incomplete Freund’s adjuvant and injected subcutaneously. Peripheral blood mononuclear cells obtained before and after vaccination by leukapheresis were analyzed using a fluorescence-based HLA/peptide-tetramer binding assay and cytokine flow cytometry.

Results: Vaccination induced an increase in peptide-specific T cells in 28 of 29 patients. The median frequency of CD8+T cells specific for the g209–2M peptide increased markedly from 0.02% before to 0.34% after vaccination (P < .0001). Eight patients (28%) exhibited peptide-specific CD8+T-cell frequencies greater than 1%, including two patients with frequencies of 4.96% and 8.86%, respectively. Interferon alfa-2b–treated patients also had significant increases in tetramer-binding cells (P < .0001). No difference was observed between the every-2-weeks and the every-3-weeks vaccination schedules (P = .59).

Conclusion: Flow cytometric analysis of HLA/peptide-tetramer binding cells was a reliable means of quantifying the CD8+T-cell response to peptide immunization. This assay may be suitable for use in future trials to optimize different vaccination strategies. Concurrent interferon treatment did not inhibit the development of a peptide-specific immune response and vaccination every 2 weeks, and every 3 weeks produced similar results.
Phase II Trial of Intravenous CI-1042 in Patients With Metastatic Colorectal Cancer
Authors: Hamid Oday , Varterasian Mary L. , Wadler Scott , Hecht J. Randolph , Benson Al , Galanis Evanthia , Uprichard Margaret , Omer Charles , Bycott Paul , Hackman Robert C. , Shields Anthony F.
Published: April 2003

Purpose: To evaluate the antitumor activity, safety, immune response, and replication of CI-1042 (ONYX-015), an E1B 55-kd gene-deleted replication-selective adenovirus, administered intravenously to patients with metastatic colorectal cancer

Patients and Methods: Eighteen patients with metastatic colorectal cancer for whom prior chemotherapy failed were enrolled onto an open-label, multicenter, phase II study. CI-1042 was administered intravenously at a dose of 2 × 1012 viral particles every 2 weeks. Patients were evaluated for tumor response and toxicity; in addition, blood samples were taken for adenovirus DNA and neutralizing antibody analysis.

Results: Common toxicities included flu-like symptoms, nausea, and emesis. All 18 patients eventually were removed from study because of progressive disease. Seven patients were assessed as having stable disease after 2 months of treatment, whereas two patients were considered to have stable disease after 4 months. Detectable circulating CI-1042 DNA was identified in 36% of patients 72 hours after last infusion, which is suggestive of ongoing viral replication.

Conclusion: In this phase II study, intravenous CI-1042 was administered safely to patients with advanced colorectal cancer. Toxicity was manageable, consisting primarily of flu-like symptoms. Stable disease was experienced by seven patients for 11 to 18 weeks.
Cell Context-Dependent Differences in the Induction of E2F-1 Gene Expression by 17β-Estradiol in MCF-7 and ZR-75 Cells
Authors: Ngwenya Sharon , Safe Stephen
Published: April 2003

17β-Estradiol (E2) induces E2F-1 gene expression in ZR-75 and MCF-7 human breast cancer cells. Analysis of the E2F-1 gene promoter in MCF-7 cells previously showed that hormone-induced transactivation required interactions between estrogen receptor α (ERα)/Sp1 bound to upstream GC-rich sites and NFYA bound to downstream CCAAT sites within the −169 to −54 region of the promoter. This same region of the E2F-1 promoter was also E2 responsive in ERα-positive ZR-75 cells; however, further analysis of the promoter showed that cooperative ERα/Sp1/NFY interactions were not necessary for hormone-induced transactivation in ZR-75 cells. The upstream GC-rich motifs (−169 to −111) are activated independently by ERα/Sp1 in ZR-75 but not MCF-7 cells, and a construct (pE2F-1jm1) containing the −122 to −54 downstream CCAAT site that bound NFYA was also E2 responsive. E2 also induced reporter gene activity in ZR-75 cells transfected with an expression plasmid for a chimeric protein containing the DNA-binding domain of the yeast GAL4 protein fused to NFYA (pM-NFYA) and a construct containing five tandem GAL4 response elements. Subsequent studies showed that hormonal activation of pE2F-1jm1 and pM-NFYA are dependent on nongenomic pathways in which E2 activates cAMP/protein kinase A. Hormone-dependent regulation of E2F-1 gene expression in ZR-75 and MCF-7 involves the same cis elements and interacting transcription factors but different mechanisms, demonstrating the importance of cell context on transactivation pathways, even among ER-positive breast cancer cell lines.