bluuebunny/crossref_metadata_2025 huggingface dataset:
Polymorphism in polypropylene fibers
Authors: Broda Jan
Published: July 2003

The structure of polypropylene fibers formed by different spinning conditions was studied. The investigations were carried out using wide‐angle X‐ray scattering (WAXS) and small‐angle X‐ray scattering (SAXS) methods. It was stated that in noncolored fibers the mesophase and the monoclinic α forms were formed. The highest content of the mesophase was observed for fibers extruded at the higher temperature and taken at the medium take‐up velocity. In fibers colored with quinacridone pigment the structure containing the monoclinic α and the trigonal β forms was obtained. The highest content of the β form occurred in fibers taken at the lowest take‐up velocity. The increase of the take‐up velocity caused a rapid drop in β form content. In fibers taken at higher velocities only the α form was formed. It was found that the content of different forms as well lamellar thickness depends on crystallization conditions, which were influenced by the formation parameters. © 2003 Wiley Periodicals, Inc. J Appl Polym Sci 89: 3364–3370, 2003
A combinatorial approach in designing hydrophilic surfaces for solid‐phase peptide synthesis
Authors: Ercole Francesca , FitzGerald Michael , Perera Senake , Ede Nicholas , Riley Phil , Campbell Rhonda
Published: July 2003

A hydrophilic surface suitable for solid‐state peptide synthesis was developed on a solid support called a lantern. The split‐and‐mix combinatorial technique was used to prepare about 500 surfaces in a very short time. Surfaces were analyzed according to values for gel formation, percentage weight grafted, grafted copolymer composition, and number of functional groups per lantern. These values were correlated to the purity of a peptide synthesized on these surfaces. © 2003 Wiley Periodicals, Inc. J Appl Polym Sci 89: 3371–3378, 2003
Crystallization kinetics and PVT behavior of poly(vinylidene fluoride) in process conditions
Authors: Pantani R. , Speranza V. , Besana G. , Titomanlio G.
Published: July 2003

The so‐called fluoropolymers have gained, in recent years, considerable industrial success, and the increasing industrial interest in this class of materials has caused a need for better characterization of the properties of interest for processability, for instance, for injection molding or extrusion. In this work, the pressure–volume–temperature (PVT) relationship of a poly(vinylidene fluoride) is described by combining specific volumes of amorphous and crystalline phases present in the material. The volumes of the two phases are described simply by thermal expansion and compressibility coefficients drawn from standard PVT data below and above the crystallization range. Within the crystallization range, the material volume is assumed to change from amorphous to crystalline according to the evolution of an overall crystallinity degree, which is described by the Nakamura nonisothermal formulation of an Avrami crystallization kinetic model. Model parameters are identified by comparison with standard calorimetric results, PVT data, and final densities of thin samples solidified during quenches conducted with cooling rates of several hundreds of Kelvins/second. The resulting model allows the description of the PVT behavior of PVDF in the pressure ‐and cooling‐rate ranges of interest for processing. © 2003 Wiley Periodicals, Inc. J Appl Polym Sci 89: 3396–3403, 2003
Oligophrenin‐1, a Rho GTPase‐activating protein (RhoGAP) involved in X‐linked mental retardation, is expressed in the enteric nervous system
Authors: Xiao Junhua , Neylon Craig B. , Hunne Billie , Furness John B.
Published: July 2003

Oligophrenin‐1 is a RhoGTPase‐activating protein (RhoGAP) that is involved in the regulation of shape changes in dendritic spines, and outgrowth of axons and dendrites in the brain. These changes in neuronal morphology are central to the mechanisms of plasticity, learning, and memory. Although the enteric nervous system also exhibits long‐term changes in neuronal function, the expression and involvement of oligophrenin‐1 has not previously been investigated. We show by RT‐PCR analysis that oligophrenin‐1 mRNA is expressed in the myenteric plexus (MP) of the guinea pig ileum. Sequencing of RT‐PCR products showed that guinea pig oligophrenin‐1 mRNA is 98% and 87% homologous to human and mouse oligophrenin‐1, respectively, except that a 42 bp sequence is absent from the guinea pig mRNA. This 42 bp sequence codes for a sequence of 14 amino acids located near the carboxy‐terminal end of the RhoGAP domain in the human sequence. An antibody that recognizes human oligophrenin‐1 identified a 91 kDa protein band in rat and mouse brain lysates and in guinea pig sciatic nerve, and a 36 kDa protein band in both purified enteric ganglion cell and brain lysate from guinea pig. Oligophrenin‐1 is localized specifically to neurons and varicose axons in the MPs and submucosal plexuses (SMPs) of the guinea pig and rat, but is not detectable in glial cells, smooth muscle, or other cell types. These findings indicate that oligophrenin‐1 is expressed in the enteric nervous system, where it may regulate morphological changes in axons and dendrites, and thus modulate neuronal connectivity. Anat Rec Part A 273A:671–676, 2003. © 2003 Wiley‐Liss, Inc.
Effect of lacunocanalicular architecture on hydraulic conductance in bone tissue: Implications for bone health and evolution
Authors: Mishra Sanjay , Knothe Tate Melissa L.
Published: July 2003

Bone tissue health depends largely on efficient fluid and solute transport between the blood supply and cells that are the living component of the tissue. We hypothesized that the lacunocanalicular hydraulic network, which is defined by the pericellular fluid space that is common to all bone tissue, is optimized to transport fluid and solutes between the blood supply and bone cells. An analytical study was carried out to evaluate the effect of osteonal architecture, including the osteon diameter, number of annular lamellar regions, and number and length of canalicular channels, on fluid transport between the blood supply and bone cells. On the basis of this analysis, we conclude that osteon size is limited to the distance over which fluid and solutes can be transported efficiently between the blood supply and cells. This analytic model suggests that hydraulic conductivity is highest in lamellar regions closest to the Haversian canal (HC) and decreases with increasing distance from the blood supply, reaching a plateau after the fifth lamella (169 μm radius). Furthermore, an increase in the diameter of the HC, or a decrease in the length of canaliculi, reduces the hydraulic conductivity within the lacunocanalicular network. Applying the principle of minimal expenditure of energy to this analysis, the path distance comprising five or six lamellar regions represents an effective limit for fluid and solute transport between the blood supply and cells; beyond this threshold, hydraulic resistance in the network increases and additional energy expenditure is necessary for further transportation. This suggests that transport is optimized to meet metabolic demands concomitant with a minimal expenditure of energy. This fundamental new insight into bone structure and physiology may provide a new basis of understanding for tissue engineering, bone physiology in health and disease, and evolutionary biology. Anat Rec Part A 273A:752–762, 2003. © 2003 Wiley‐Liss, Inc.
Histological studies of bone formation during pedicle restoration and early antler regeneration in roe deer and fallow deer
Authors: Kierdorf Uwe , Stoffels Elke , Stoffels Dieter , Kierdorf Horst , Szuwart Thomas , Clemen Günter
Published: July 2003

The purpose of the present study was to examine the process of bone formation in the regenerating cranial appendages of roe deer (Capreolus capreolus) and fallow deer (Dama dama) during the early postcasting period. After the antlers are cast, osteoclastic and osteoblastic activities lead to a smoothing of the pedicle's separation surface, a strengthening of the pedicle bone, and a partial restoration of the distal pedicle portion that was lost along with the cast antler. Initially, bone formation occurs by intramembranous ossification, but early during the regeneration process cartilage is formed at the tips of the cranial appendages, and is subsequently replaced by bone in a process of endochodral ossification. Shortly after the antlers are cast, the cambium layer of the periosteum in the distal pedicle is markedly enlarged, which suggests that the periosteum serves as a cell source for the bone‐forming tissue covering the exposed pedicle bone. The histological findings of our study are consistent with the view that the bony component of the regenerating cranial appendages of deer is largely derived from the pedicle periosteum. Based on findings in other bone systems, we speculate that stem cells that can undergo both osteogenic and chondrogenic differentiation are present in the pedicle periosteum. The early onset of chondrogenesis in the regeneration process is regarded as an adaptation to the necessity of producing a huge volume of bone within a short period. This parallels the situation in other cases of chondrogenesis in membrane bones. Anat Rec Part A 273A:741–751, 2003. © 2003 Wiley‐Liss, Inc.
Immunolocalization of calbindin D28k and vitamin D receptor during root formation of murine molar teeth
Authors: Onishi Tomoyuki , Okawa Rena , Murakami Hiroaki , Ogawa Tomohiro , Ooshima Takashi , Wakisaka Satoshi
Published: July 2003

Cells in the epithelial rest of Malassez (ERM cells) express calbindin D28k (CB); however, the hormonal regulation of CB in ERM cells remains to be elucidated. We investigated the immunohistochemical localization of CB and 1,25‐dihydroxyvitamin D3 receptor (VDR) during root formation of mouse molar teeth in order to clarify whether the expression of CB in ERM cells is dependent on vitamin D. At the early stage of root formation (postnatal (PN) days 10–14), both CB‐ and VDR‐immunoreactive cells were observed intermittently along the root surface. In the apical portion, almost all CB‐immunoreactive cells showed VDR immunoreactivity; however, VDR‐immunoreactive cells in the most apical portion were immunonegative for CB. In the middle and cervical portions, the distributions of the two proteins were completely different. At the late stage of root formation (PN28d) and in adult animals, CB immunoreactivity was distributed in cells found along the acellular cementum at the bifurcation region, as well as between the dentin and cellular cementum in the apical portion (although these lacked immunoreactivity for VDR). The present results indicate that CB expression in newly disrupted cells from Hertwig's epithelial root sheath occurs in a vitamin‐D dependent manner, whereas the expression of CB in mature ERM cells may be independent of vitamin D. Anat Rec Part A 273A:700–704, 2003. © 2003 Wiley‐Liss, Inc.
Immortalization of normal human embryonic fibroblasts by introduction of either the human papillomavirus type 16 E6 or E7 gene alone
Authors: Yamamoto Akito , Kumakura Shin‐ichi , Uchida Minoru , Barrett J. Carl , Tsutsui Takeki
Published: July 2003

The ability of the human papillomavirus type 16 (HPV‐16) E6 or E7 gene to induce immortalization of normal human embryonic fibroblast WHE‐7 cells was examined. WHE‐7 cells at 9 population doublings (PD) were infected with retrovirus vectors encoding either HPV‐16 E6 or E7 alone or both E6 and E7 (E6/E7). One of 4 isolated clones carrying E6 alone became immortal and is currently at >445 PD. Four of 4 isolated clones carrying E7 alone escaped from crisis and are currently at >330 PD. Three of 5 isolated clones carrying E6/E7 were also immortalized and are currently at >268 PD. The immortal clone carrying E6 only and 2 of the 3 immortal clones carrying E6/E7 expressed a high level of E6 protein, and all the immortal clones carrying E7 alone and the other immortal clone carrying E6/E7 expressed a high level of E7 protein when compared to their mortal or precrisis clones. The immortal clones expressing a high level of E6 or E7 protein were positive for telomerase activity or an alternative mechanism of telomere maintenance, respectively, known as ALT (alternative lengthening of telomeres). All the mortal or precrisis clones were negative for both phenotypes. All the immortal clones exhibited abrogation of G1 arrest after DNA damage by X‐ray irradiation. The expression of INK4a protein (p16INK4a) was undetectable in the E6‐infected mortal and immortal clones, whereas Rb protein (pRb) was hyperphosphorylated only in the immortal clone. The p16INK4a protein was overexpressed in all the E7‐infected immortal clones and their clones in the pre‐crisis period as well as all the E6/E7‐infected mortal and immortal clones, but the pRb expression was downregulated in all of these clones. These results demonstrate for the first time to our knowledge that HPV‐16 E6 or E7 alone can induce immortalization of normal human embryonic fibroblasts. Inactivation of p16INK4a/pRb pathways in combination with activation of a telomere maintenance mechanism is suggested to be necessary for immortalization of normal human embryonic fibroblasts by these viral oncogenes. The susceptibility of human cells to immortalization may be related to the state of differentiation of the cells. © 2003 Wiley‐Liss, Inc.
Overexpression of thymidylate synthase mediates desensitization for 5‐fluorouracil of tumor cells
Authors: Saga Yasushi , Suzuki Mitsuaki , Mizukami Hiroaki , Kohno Takahiro , Takei Yuji , Fukushima Masakazu , Ozawa Keiya
Published: July 2003

The prognosis of cancers of various organs overexpressing thymidylate synthase (TS) has been reported to be poor. It has been suggested that the poor prognosis is partly due to a low sensitivity of TS‐overexpressing tumors to TS‐targeting 5‐fluorouracil (5‐FU). To investigate the relationship between TS expression and sensitivity to 5‐FU, we used the TS‐overexpressing cervical cancer cell line SKG‐II/TS and SKG‐I/TS that had been established by TS gene transfer. The 50% growth inhibitory concentration (IC50) of 5‐FU for SKG‐II/TS was 24 ± 6.0 μM, which was 6 times as high as that for the control (4.0 ± 1.1 μM), showing significantly decreased sensitivity to 5‐FU (p < 0.01). The IC50 of 5‐FU for SKG‐I/TS was 90 ± 15 μM, which was over 2 times as high as that for the control (40 ± 0.6 μM), showing significantly decreased sensitivity to 5‐FU (p < 0.05). Thus, TS‐overexpressing tumors have decreased sensitivity to 5‐FU, which may be one of the factors that determine the prognosis of these tumors. © 2003 Wiley‐Liss, Inc.
Cohort studies of association between self‐reported allergic conditions, immune‐related diagnoses and glioma and meningioma risk
Authors: Schwartzbaum Judith , Jonsson Fredrik , Ahlbom Anders , Preston‐Martin Susan , Lönn Stefan , Söderberg Karin C. , Feychting Maria
Published: July 2003

An inverse association between self‐reported allergies and glioma and meningioma risk, has been previously observed in case‐control studies. Approximately 27% (median) of the information on both glioma and meningioma in these studies, however, is collected from proxy respondents. In fact, the odds ratios (OR) among previous brain tumor studies are inversely related to the proportion of proxy respondents (Pearson correlation coefficient = −0.94; 95% CI = −1.00 to −0.65); this correlation suggests bias. We therefore constructed 3 cohorts based on the Swedish Twin, Hospital Discharge, and Cancer Registries. In Cohorts I (14,535 people developed 37 gliomas and 41 meningiomas) and II (29,573 people developed 42 gliomas and 26 meningiomas) median time from self‐report of allergies to brain tumor diagnosis was 15.4 years. Cohort III, which overlaps with Cohorts I and II (52,067 people developed 68 gliomas and 63 meningiomas), was linked to the Swedish Hospital Discharge Registry where pre‐brain tumor immune‐related discharge diagnoses were recorded. Allergies are inversely associated with glioma risk in Cohort I (Hazard ratio [HR] = 0.45; 95% CI = 0.19–1.07) and among high grade (III and IV, HR = 0.45; 95% CI = 0.11–1.92) but not low grade (I and II, HR = 2.60; 95% CI = 0.86–7.81) gliomas in Cohort II. In Cohort III, immune‐related discharge diagnoses are also inversely associated with glioma (HR = 0.46; 95% CI = 0.14–1.49). There is no strong evidence against (and some for) the hypothesis that allergies reduce glioma risk. © 2003 Wiley‐Liss, Inc.
A common 936 C/T gene polymorphism of vascular endothelial growth factor is associated with decreased breast cancer risk
Authors: Krippl Peter , Langsenlehner Uwe , Renner Wilfried , Yazdani‐Biuki Babak , Wolf Gerald , Wascher Thomas C. , Paulweber Bernhard , Haas Josef , Samonigg Hellmut
Published: July 2003

A common 936 C/T polymorphism in the gene for the vascular endothelial growth factor (VEGF) has been associated with VEGF plasma levels. In our case‐control study, we investigated the role of this polymorphism for breast cancer risk. VEGF genotype was determined in 500 women with breast cancer and 500 sex‐ and age‐matched healthy control subjects. Carriers of a 936T‐allele were more frequent among controls (29.4%) than among patients (17.6%; p = 0.000014). The odds ratio for carriers of a 936T‐allele for breast cancer was 0.51 (95% confidence interval 0.38–0.70). Additionally, VEGF plasma levels were determined in 21 nonsmoking post‐menopausal controls; carriers of a 936T allele had significantly lower levels (median 23 pg/ml; range 6–50 pg/ml) than noncarriers (37; 21–387; p = 0.034). We conclude that carriers of a VEGF 936T‐allele are at decreased risk for breast cancer, this, however, requiring further confirmation in a larger study. © 2003 Wiley‐Liss, Inc.
Test characteristics of visual inspection with 4% acetic acid (VIA) and Lugol's iodine (VILI) in cervical cancer screening in Kerala, India
Authors: Sankaranarayanan Rengaswamy , Wesley Ramani , Thara Somanathan , Dhakad Namrata , Chandralekha Bharathykutty , Sebastian Paul , Chithrathara K. , Parkin Donald Maxwell , Nair Madhavan Krishnan
Published: July 2003

Simple and inexpensive methods based on visual examination of the cervix are currently being investigated as alternative methods of cervical screening. The test characteristics of visual inspection with 4% acetic acid (VIA), and Lugol's iodine (VILI) and conventional cytology were investigated in a cross‐sectional study involving 4,444 women aged 25 to 65 years in Kerala, India. While detection of any acetowhite area constituted a low‐threshold positive VIA, detection of well‐defined, opaque acetowhite lesions close to or touching the squamocolumnar junction constituted a high‐threshold positive VIA test. Detection of definite yellow iodine nonuptake areas in the transformation zone close to or touching the squamocolumnar junction constituted a positive VILI test. Cytology was considered positive if reported as atypia or worse lesions. All screened women were evaluated by colposcopy and biopsies were directed in 1,644 women (37.0%), which allowed the direct estimation of sensitivity, specificity and predictive values. The reference diagnosis was based on a combination of histology and/or colposcopy. True disease status was defined as CIN 2 and worse lesions. A total of 149 (3.4%) women had CIN 2 or worse lesions. The sensitivities of low‐threshold VIA, high‐threshold VIA, VILI and cytology to detect CIN 2 or worse disease were 88.6%, 82.6%, 87.2% and 81.9%, respectively; the corresponding specificities were 78.0%, 86.5%, 84.7% and 87.8%. Our results indicate that VIA and VILI are suitable alternate screening tests to cytology for detecting cervical neoplasia in low‐resource settings. © 2003 Wiley‐Liss, Inc.
Expression of tumor markers hyaluronic acid and hyaluronidase (HYAL1) in head and neck tumors
Authors: Franzmann Elizabeth J. , Schroeder Grethchen L. , Goodwin William Jarrard , Weed Donald T. , Fisher Penelope , Lokeshwar Vinata B.
Published: July 2003

Characteristic behaviors of head and neck squamous cell carcinoma (HNSCC) include a propensity to occur as multiple synchronous and metachronous tumors, frequent recurrence and metastasis. Early detection of HNSCC and monitoring its recurrence are necessary to improve prognosis. Hyaluronic acid (HA), a component of extracellular matrix, promotes metastasis. Small fragments of HA stimulate angiogenesis. HA fragments are generated when hyaluronidase (HAase), an endoglycosidase, degrades the HA polymer. Using the HA test (an ELISA‐like assay) we found that saliva HA levels are 4.9‐fold elevated in 11 HNSCC patients (2841 ± 887 ng/mg protein) when compared to 6 normal controls (579.3 ± 122.6 ng/mg protein; p = 0.00238). HNSCC patients included in our study were patients with cancers of the oral cavity (n = 4), pharynx (n = 7) and larynx (n = 1). The HA levels were also elevated in MDA‐1483, FaDu and HEp‐2 cell lines when compared to the transformed keratinocyte line HEK‐001. Saliva HAase levels measured using the HAase test (an ELISA‐like assay) were 3.7‐fold elevated in HNSCC patients (10.4 ± 1.4 mU/mg protein) when compared to normal controls (2.8 ± 0.7 mU/mg protein; p = 0.0028). MDA‐1483 and HEp‐2 cells secreted 7‐ to 11‐fold higher levels of HAase in their conditioned media (CM) when compared to FaDu cells, and the latter secreted 1.5‐fold more HAase than HEK‐001 cells. Reverse transcriptase (RT)‐PCR analysis detected the expression of full‐length HYAL1 type HAase transcript in tumor cells. None of the cells exhibited the expression of PH20 in RT‐PCR analysis. Immunoblot analysis confirmed the expression of a ∼55 kDa HYAL‐related protein in tumor cell CM and in patients' saliva. The pH activity profile and optimum (pH 4.4) of the HAase activity present in HNSCC patients' or normal saliva and that secreted in the CM of tumor cells closely resembled that of the partially purified HYAL1 type HAase. The profiles of HA species in HNSCC patients' and normal saliva are different. The high‐stage HNSCC patients' saliva contains a high‐molecular‐mass HA species and HA fragments, in addition to the HA species present in the normal individual's saliva. These results show that HYAL1 is the major tumor‐derived HAase expressed in HNSCC. Furthermore, HA and HAase may be sensitive and specific markers for detecting HNSCC and monitoring its recurrence. Further studies are needed to confirm these preliminary studies. © 2003 Wiley‐Liss, Inc.
Thyroid cancer risk after thyroid examination with131I: A population‐based cohort study in Sweden
Authors: Dickman Paul W. , Holm Lars‐Erik , Lundell Göran , Boice John D. , Hall Per
Published: July 2003

Ionizing radiation is the only established cause of thyroid cancer, though the effect of diagnostic administration of131I on thyroid cancer risk appears minimal. The annual number of thyroid examinations using radioiodine is currently 5 per 1,000 individuals worldwide, so this issue is of public health importance. Our objective was to evaluate the excess risk of thyroid cancer following a range of known doses of131I administered for diagnostic purposes. We conducted a nationwide, population‐based cohort study in Sweden including all 36,792 individuals who received131I for diagnostic purposes during 1952–1969 and were alive and free of thyroid cancer 2 years after exposure. Accrual of person‐time at risk commenced 2 years after the first131I administration. Follow‐up for cancer was to the end of 1998. Standardized incidence ratios (SIRs) were calculated as the ratio between the observed and expected numbers of thyroid cancers. Estimates were stratified by previous exposure to external radiation therapy to the neck, reason for thyroid examination,131I dose, sex, age at exposure and time since exposure. Thyroid cancers (n= 129) were diagnosed during 886,618 person‐years at risk. Excess thyroid cancers were observed only among the 1,767 patients who reported previous external radiation therapy to the neck [SIR = 9.8, 95% confidence interval (CI) 6.3–14.6] and among those originally referred due to suspicion of a thyroid tumor (SIR = 3.5, 95% CI 2.7–4.4 for 11,015 patients without previous external radiation therapy). The 24,010 patients without previous exposure to external radiation therapy to the neck who were referred for a reason other than suspicion of a thyroid tumor received an estimated dose to the thyroid of 0.94 Gy. Among these patients, 36 thyroid cancers were observed compared to 39.5 expected (SIR = 0.91, 95% CI 0.64–1.26). We found no evidence that administration of131I for diagnostic purposes increases risk of thyroid cancer. However, our study included few patients under age 20, so the results apply primarily to exposure among adults. Our data suggest that protraction of dose may result in a lower risk than brief X‐ray exposure of the same total dose. © 2003 Wiley‐Liss, Inc.
Human papillomavirus positivity predicts favourable outcome for squamous carcinoma of the tonsil
Authors: Li Wei , Thompson Carol H. , O'Brien Christopher J. , McNeil Edward B. , Scolyer Richard A. , Cossart Yvonne E. , Veness Michael J. , Walker D. Murray , Morgan Gary J. , Rose Barbara R.
Published: July 2003

Mutations in the p53 and retinoblastoma (pRb) pathways associated with the use of tobacco and alcohol are common in squamous cell carcinoma (SCC) of the head and neck. Cell cycle proteins are also affected by human papillomavirus (HPV), which may also have an aetiological role in cancers at particular sites, most notably the tonsil. Attempts to identify prognostic molecular markers in head and neck cancers have met with conflicting results, but few studies have been undertaken with tumours of known HPV status at a single anatomic site. In our study 86 tonsil cancers were analysed for HPV status by sequence analysis of polymerase chain reaction products and for the expression of cell cycle proteins (p53, p21CIP1/WAF1, pRb, p16INK4A, cyclin D1 and p27KIP1) by immunohistochemistry. The HPV status could be established in 67 of the tumours. Thirty‐one (46%) of these were HPV‐positive, predominantly (28/31) for HPV16. Findings were related to tumour recurrence and patient survival. None of the cell cycle proteins independently predicted recurrence or survival. Patients with HPV‐positive tumours, however, were significantly less likely (p< 0.05) to have recurrence or to die of disease than those with HPV‐negative tumours, after adjusting for the effects of the cell cycle proteins, clinical stage, pathological node status, tumour grade, age, gender and treatment. These findings support the concept that HPV‐positive tonsil cancers may be a distinct biological group with less aggressive characteristics. Screening of tonsil cancers for HPV DNA may help optimise treatment and provide more accurate prognostic information. © 2003 Wiley‐Liss, Inc.
Analysis of the transcription regulator, CNOT7, as a candidate chromosome 8 tumor suppressor gene in colorectal cancer
Authors: Flanagan James , Healey Sue , Young Joanne , Whitehall Vicki , Chenevix‐Trench Georgia
Published: July 2003

Loss of heterozygosity (LOH) on the short arm of chromosome 8 occurs at high frequencies in many tumor types, including colorectal carcinoma. We have previously used microcell‐mediated chromosome transfer (MMCT) to map an approximately 7.7 Mb colorectal cancer suppressor region (CRCSR) at 8p22‐23.1. We have now taken a candidate gene approach to identify the putative tumor suppressor gene located within the CRCSR. CNOT7 encodes a subunit of the CCR4‐Not transcription complex and is located at 8p22. We showed that CNOT7 is expressed in normal colonic mucosa and in colonic crypt cells, as well as in colorectal cell lines and primary tumors. We assembled a panel of 88 primary colorectal tumors comprising 20 MSI‐high (high microsatellite instability), 19 MSI‐low and 49 MSS (microsatellite stable) tumors for mutation analysis of the CNOT7 gene. Denaturing high‐performance liquid chromatography (DHPLC) analysis of the entire coding region of the CNOT7 gene revealed only one somatic missense mutation in an MSS tumor. The rarity of somatic mutations in CNOT7, and its expression in primary colorectal tumors and cell lines, suggests that CNOT7 is not the target tumor suppressor gene in the 8p22‐23.1 CRCSR. © 2003 Wiley‐Liss, Inc.
Baicalein and baicalin are potent inhibitors of angiogenesis: Inhibition of endothelial cell proliferation, migration and differentiation
Authors: Liu Jun‐Jen , Huang Tien‐Shang , Cheng Wen‐Fang , Lu Fung‐Jou
Published: July 2003

In recent studies, we have shown that baicalein and baicalin, 2 major flavonoids of Scutellaria baicalensis Georgi, exhibit anticancer activity against several cancers in vitro. In our present study, we assessed their potential as anti‐angiogenic agents in vivo employing chicken chorioallantoic membrane (CAM) assay and in vitro human umbilical vein endothelial cells (HUVECs) culture. When CAMs were treated with either baicalein or baicalin for 48 hr, the angiogenic response induced by basic fibroblast growth factor (bFGF) was markedly reduced in a dose‐dependent manner. Further characterization showed that both flavonoids exhibited dual antiproliferative (at low dose) and apoptogenic (at high dose) effects on HUVECs. In biochemical analysis, treatment of HUVECs with baicalein and baicalin for 24 hr resulted in a dose‐dependent decrease in the matrix metalloproteinase (MMP)‐2 activity. Moreover, the migration of endothelial cells and the differentiation of endothelial cells into branching networks of tubular structures in vitro were also inhibited by these 2 flavonoids in a dose‐dependent manner. Baicalein is more potent than baicalin in anti‐angiogenesis in vivo as well as in vitro. Taken together, the results of our study provide evidence that baicalein and baicalin possess an anti‐angiogenesis potential that is a previously unrecognized biologic activity. © 2003 Wiley‐Liss, Inc.
Increased in situ gelatinolytic activity in renal cell tumor tissues correlates with tumor size, grade and vessel invasion
Authors: Kamiya Naoto , Kishimoto Takashi , Suzuki Hiroyoshi , Sekita Nobuyuki , Nagai Yuichiro , Oosumi Nobuyuki , Kito Hiroki , Tochigi Naobumi , Shinbo Masataka , Nemori Ryouichi , Ichikawa Tomohiko , Igarashi Tatsuo , Ito Haruo , Ishikura Hiroshi
Published: July 2003

Degradation of collagen, or gelatinolysis, by tumor cells is one of the most important events in tumorigenesis. We investigate the possible relationship between the in situ gelatinolytic activities exerted by matrix metalloproteinases (MMPs) and clinico‐pathological factors in renal cell tumor (RCT) patients. Using the film in situ zymography (FIZ) method, we determined in situ localization of MMP‐like gelatinolytic activities in cancerous and normal tissues in the kidney (n = 51). To clarify the MMP(s) responsible for the gelatinolytic activity in RCTs, we examined the expressions of MMP‐2 and MMP‐9 in the kidney tissues by means of gelatin zymography (GZG). MMP expression was also detected by RT‐PCR and Western blotting analysis. We then investigated the associations of MMP expression, as detected by GZG, with the intensity of gelatinolytic activity, as determined by FIZ. We analyzed the possible relationship of FIZ findings to several clinico‐pathological factors such as tumor size, grade, vessel invasion, histologic type, stage and metastasis. FIZ demonstrated that all tumor and normal kidney tissues showed in situ gelatinase activities, and that gelatinolytic activities in RCTs were much stronger than those of normal kidney tissues. There was a statistically significant correlation between the intensity of MMP‐like gelatinolytic activity and tumor size, tumor grade and vessel invasion (p < 0.05), but not between it and histological type, tumor stage or metastatic status. FIZ showed that tumor tissues in 5 of the 6 patients with fatal outcome exhibited the intense gelatinolytic pattern. Stronger in situ gelatinolytic patterns were documented in cases with higher MMP‐2 expression. The molecular species of MMPs detected by GZG were confirmed by RT‐PCR and Western blotting analysis. The FIZ technique enables a direct assessment of in situ gelatinolytic activity in RCT tissues. The intensity of the activity seems to affect the biology of RCT tissues. Our results also indicate a major role for MMP‐2 in in situ gelatinolysis in RCT tissues. © 2003 Wiley‐Liss, Inc.
Mapping protein expression in mouse pancreatic islets by immunolabeling and electron energy loss spectrum‐imaging
Authors: Goping Gertrud , Pollard Harvey B. , Srivastava Meera , Leapman Richard
Published: July 2003

A combination of immuno‐electron microscopy and electron energy‐loss spectrum‐imaging was used to map the distributions of endocrine polypeptide hormones and proteins in mouse pancreatic islet of Langerhans. Tissue was analyzed from control animals and from mice that were heterozygous for the Anx7 gene, which defines a Ca2+/GTP‐dependent membrane fusion and ion channel protein. The heterozygous Anx7 (+/−) mouse displays defects in IP3 receptor mediated Ca2+ signaling and insulin secretion. Therefore, information was obtained about the distributions of the hormones insulin and glucagon, as well as the proteins ANX7 and the IP3 receptor. Insulin secretion appears to be defective in the mutants. It was found from immunolabeling experiments that expression of the IP3 receptor is reduced in mutant islets compared to control islets. Subcellular distributions of sulfur and nitrogen obtained by electron energy‐loss spectrum‐imaging showed that the insulin concentrations of β granules were essentially the same in control and mutant islets. By contrast, immunogold labeling of mutant islets shows more insulin immunoreactivity in the β granules. It follows that insulin may be packaged differently in mutant islets, making antigenic determinants more available to the labeling antibody. The increased rate of insulin secretion in the hyperplastic mutant islets can be explained by compensatory increases in islet size, rather than by an increased insulin concentration in the β cells. The results indicate that reduced ANX7 expression leads to defects in the IP3 receptor expression in the endocrine cells of the mutant mouse. Increased size of the islet or of adrenal medulla may be a compensatory mechanism for secretion defect by individual endocrine cells. Defects in IP3 receptor expression, and documented consequences of a Ca2+ signaling defect, lead to other changes in organelles such as the mitochondrial number in islet β‐cells. The effects and consequences of reduced ANX7 expression on mitochondria are evident in ultrastructural observations. Microsc. Res. Tech. 61:448–456, 2003. Published 2003 Wiley‐Liss, Inc.
Ultrastructural changes in feline dental pulp with periodontal disease
Authors: Ghoddusi Jamileh
Published: July 2003

A light and transmission electron microscopic study was conducted on dental pulp on cats suffering periodontal disease. After extraction, pulp tissues were fixed and embedded in Epon‐Araldite. Thick layers of predentin (50 μm) and odontoblasts (30 μm) were observed. In thin sections, odontoblasts showed many mitochondria and secretary vesicles. Some capillaries with several fenestrations were located within the odontoblastic layer. All the sections of pulp examined displayed a generalized infiltration of chronic inflammatory cells. Fibroblasts displayed lytic changes in some areas. These findings imply that the pulp is significantly affected by periodontal disease and furcation‐involved teeth should be a carefully considered factor when dental treatment is planned. Microsc. Res. Tech. 61:423–427, 2003. © 2003 Wiley‐Liss, Inc.
Changes in particle area measurements due to SEM accelerating voltage and magnification
Authors: Yañez María Julia , Barbosa Silvia Elena
Published: July 2003

Scanning electron microscopy (SEM) images of polymer blends followed by digital image analysis is a rapid and easy method for the measurement of particle size and dispersion. The particle size determination is done with appropriate off‐line image analysis software. However, it is necessary to understand how machine parameters involved in the formation of the SEM image influence area measurements of morphological features. In this work, the influence of the accelerating voltage used during image acquisition was examined with standard samples and with polymer blend samples. A systematic study centered on two mutually exclusive assumptions of area variation or no area variation with accelerating voltage was carried out. The off‐line image analysis software was then calibrated according to the assumptions. The main conclusion of this study was that kV has an important influence on area measurement in SEM images. This effect was observed for different standard materials (metallic and polymeric) and for the range of magnifications used. The higher the accelerating voltages, the greater the error at high magnification for polymer samples. As the beam energy increases, the primary electrons penetrate more deeply into the solid specimen, producing low‐resolution signals. These signals degrade the image and surface details, which became less well defined. Therefore, images of polymer samples must be taken at lower accelerating voltages so the desired surface details can be imaged clearly. To avoid area measurement errors, particle measurement must be done with the calibration of the off‐line image analysis software corresponding to the accelerating voltage and magnification used for the acquired images. Microsc. Res. Tech. 61:463–468, 2003. © 2003 Wiley‐Liss, Inc.
Morphological abstraction of thyroid tumor cell nuclei using morphometry with factor analysis
Authors: Murata Shin‐Ichi , Mochizuki Kunio , Nakazawa Tadao , Kondo Tetsuo , Nakamura Nobuki , Yamashita Hiroto , Urata Yoji , Ashihara Tsukasa , Katoh Ryohei
Published: July 2003

Various morphonuclear studies by digital image analysis have successfully been applied to quantify the nuclear morphology, including chromatin distribution pattern, in cytology of various organs; however, the majority of past reports have not shown correlation between the quantitative data by digital image analysis and cytological findings in practical diagnosis. In this report, we present the usefulness of morphological abstraction to combine the objective data and subjective observation in cytological diagnosis. Randomly selected, 100 cells in each Papanicolaou‐stained ABC smear samples of 39 benign and malignant thyroid tumor cases were studied. Gray‐level image data provided seven parameters for nuclear size, four parameters for nuclear shape, and 16 parameters showing the nuclear chromatin patterns from high‐dimensional texture analysis of using co‐occurrence and run‐length matrices. To statistically abstract nuclear morphology, factor analysis was used. Factor analysis classified morphological nuclear characters as abstraction parameter into five abstract parameters composed of nuclear size, shape, heterogeneity, and contrast and homogeneity of chromatin pattern. The nuclei of papillary carcinoma showed larger size, more irregular shape, and higher contrast of chromatin pattern than those of the benign group. The follicular carcinomas have larger nucleus in each cell and more monotonous chromatin pattern among cells in each case than those of the benign group. Morphological abstraction by morphometry with factor analysis may provide a practical approach to the detection of the underlying characteristics of nuclear morphology in aspiration biopsy cytology. Microsc. Res. Tech. 61:457–462, 2003. © 2003 Wiley‐Liss, Inc.
Applications of environmental scanning electron microscopy (ESEM) in biomaterials field
Authors: Manero J.M. , Gil F.J. , Padrós E. , Planell J.A.
Published: July 2003

Various tasks were undertaken in our laboratory where environmental scanning electron microscopy (ESEM) has been of particular interest within the biomaterials field. The possibility of observing wet samples, as well as the fact that sample preparation is minimal, has improved shorter time scales and lower costs in microscopy. Minimal preparation has also reduced the possibility of introducing artifacts. Examples like cell cultures used for pit resorption assays, calcium phosphate deposition processes, and dissolution of phosphate glasses used as biomaterials are presented. Finally, a servohydraulic testing machine designed for mechanical testing in situ in ESEM has allowed the study of shape memory alloys for orthodontic applications or the behavior of different adhesives used in odontology. Microsc. Res. Tech. 61:469–480, 2003. © 2003 Wiley‐Liss, Inc.
The impact of information and communications technology on managerial practices: the use of codes of conduct
Authors: Healy Mike , Iles Jennifer
Published: July 2003

The widespread adoption of advanced information and communications technology (ICT) is generally regarded as a powerful driver for undermining bureaucratic management. ICT‐enabled organizations are often associated with flattened organizational structures, decentralization of resources and activities, teamwork and the empowerment of employees.

Those lines of argument are questioned with reference to knowledge management, security, the development and implementation of intranets and intranets and the findings of surveys carried out over a period of 3 years on the use of codes of conduct governing the use of ICT by employees in a wide range of organizations.

Evidence is presented that the increasing use of Internet technology has not been accompanied by the expected move away from bureaucratic management but rather by increasing employee regulation and discipline, motivated largely by management concerns about the security and efficient operation of information systems.

Copyright © 2003 John Wiley & Sons, Ltd.
Cyclin E overexpression and amplification in human tumours
Authors: Schraml Peter , Bucher Christoph , Bissig Heidi , Nocito Antonio , Haas Philippe , Wilber Kim , Seelig Steven , Kononen Juha , Mihatsch Michael J , Dirnhofer Stefan , Sauter Guido
Published: July 2003

Cyclin E amplification and overexpression have recently been described in several tumour types. However, many tumour entities have never been examined for cyclin E alterations. Numerous and time‐consuming experiments were previously required to determine the significance of potential oncogenes across different tumour types. To overcome this problem, tissue microarrays (TMAs) consisting of 3670 primary tumours from 128 different tumour types, 709 metastases, and 354 normal tissues were generated. Cyclin E alterations were then analysed by fluorescence in situ hybridization (FISH) and immunohistochemistry (IHC). Cyclin E gene amplification was observed in 15 different tumour types and subtypes, ie rhabdomyosarcoma, urinary bladder cancer (three subtypes), ovarian cancer (two subtypes), malignant fibrous histiocytoma, adenocarcinoma of the small intestine, medullary breast cancer, gall bladder adenocarcinoma, phaeochromocytoma, gastric adenocarcinoma, squamous cell carcinoma of the uterine cervix, colonic adenocarcinoma, and endometrial carcinoma. Cyclin E protein accumulation was found in 48 different tumour types. The use of TMA technology has enabled us to expand considerably our knowledge of cyclin E alterations in human tumours. The occurrence of amplification and overexpression in many different tumour types suggests that cyclin E plays an important role in tumour biology. Copyright © 2003 John Wiley & Sons, Ltd.
Molecular characterization of Patched‐associated rhabdomyosarcoma
Authors: Kappler Roland , Calzada‐Wack Julia , Schnitzbauer Udo , Koleva Milena , Herwig Astrid , Piontek Guido , Graedler Florian , Adamski Jerzy , Heinzmann Ulrich , Schlegel Jürgen , Hemmerlein Bernhard , Quintanilla‐Martinez Leticia , Hahn Heidi
Published: July 2003

Mutations in the human homologue of Drosophila Patched1 (PTCH1) have been found in several common tumours including basal cell carcinoma, medulloblastoma, and rhabdomyosarcoma (RMS). Medulloblastoma and RMS are also present in the murine model for Ptch1 deficiency. Tumours in heterozygous Ptch1neo67/+ mice consistently exhibit elevated transcript levels of the proto‐oncogene Gli1, of Ptch1 itself, and of the insulin‐like growth factor 2 (Igf2). The present study has investigated additional molecular changes in RMSs of Ptch1 mutant mice by means of microarray analysis and protein expression analysis. The data show activation of the cell survival‐promoting Akt/protein kinase B (Pkb). Furthermore, RMSs express increased levels of the anti‐apoptotic protein Bcl‐2 and of genes and proteins known to inhibit cell proliferation, including Gadd45a and p27kip1. Taken together, the data suggest that the formation of RMSs in Ptch1 mutants is associated with the ability of tumour cells to resist apoptosis. Copyright © 2003 John Wiley & Sons, Ltd.
Discriminating expression of differentiation markers evolves in transplants of benign and malignant human skin keratinocytes through stromal interactions
Authors: Tomakidi Pascal , Stark Hans‐Juergen , Herold‐Mende Christel , Bosch Franz Xaver , Steinbauer Heinrich , Fusenig Norbert E. , Breitkreutz Dirk
Published: July 2003

Accumulating evidence indicates a decisive role for the adjacent stroma in tumour growth and dissemination. However, it is not clear how far altered differentiation such as expression of aberrant keratins and vimentin, common in invasive human carcinomas, may reflect intrinsic cell properties or a response to the tumour environment. We have addressed this by transplanting benign and malignant human HaCaT‐ras keratinocytes, seeded on collagen matrix, onto nude mice. Initially, epithelia derived from benign and malignant cells, being separated from host stroma by collagen, were poorly organized and exhibited the same differentiation markers, as identified by immunofluorescence and in situ hybridization. Epidermal basal and suprabasal keratins were expressed persistently even upon contact with newly formed stroma and malignant cell invasion. In contrast, non‐epidermal keratins (K4/K13, K8/18, K19), which were similarly synthesized by benign and malignant cells in culture and in early transplants, were differentially regulated with increasing stromal vicinity. While both proteins and mRNAs were downregulated in benign epithelia, the malignant, invasive tumour cells continuously expressed these non‐epidermal keratins throughout (K19), suprabasally (K4/13) or at invasive sites (K8/18). Furthermore, the mesenchymal protein vimentin was expressed de novo in invasive areas confronting tumour stroma. Thus, atypical tissue markers, similarly synthesized in isolated cells in vitro, are downregulated in benign but maintained and upregulated in malignant epithelia. This is presumably caused by the neighbouring stroma being permanently activated by malignant epithelia. Copyright © 2003 John Wiley & Sons, Ltd.
Extracellular matrix remodelling: the role of matrix metalloproteinases
Authors: Stamenkovic Ivan
Published: July 2003

Matrix metalloproteinases (MMPs) are a growing family of metalloendopeptidases that cleave the protein components of the extracellular matrix and thereby play a central role in tissue remodelling. For many years following their discovery, MMPs were believed to function primarily as regulators of ECM composition and to facilitate cell migration simply by removing barriers such as collagen. It is becoming increasingly clear, however, that MMPs are implicated in the functional regulation of a host of non‐ECM molecules that include growth factors and their receptors, cytokines and chemokines, adhesion receptors and cell surface proteoglycans, and a variety of enzymes. MMPs therefore play an important role in the control of cellular interactions with and response to their environment in conditions that promote tissue turnover, be they physiological, such as normal development, or pathological, such as inflammation and cancer. This review summarizes some of the recent discoveries that have shed new light on the role of MMPs in physiology and disease. Copyright © 2003 John Wiley & Sons, Ltd.