bluuebunny/crossref_metadata_2025 huggingface dataset:
Transient gene expression: Recombinant protein production with suspension‐adapted HEK293‐EBNA cells
Authors: Meissner Petra , Pick Horst , Kulangara Alexandra , Chatellard Philippe , Friedrich Kirstin , Wurm Florian M.
Published: August 2002

Transient gene expression (TGE) in mammalian cells at the reactor scale is becoming increasingly important for the rapid production of recombinant proteins. We improved a process for transient calcium phosphate‐based transfection of HEK293‐EBNA cells in a 1–3 L bioreactor volume. Cells were adapted to suspension culture using a commercially available medium (BioWhittaker, Walkersville, MD). Process parameters were optimized using a plasmid reporter vector encoding the enhanced green fluorescent protein (EGFP/CLONTECH, Palo Alto, CA, USA). Using GFP as a marker‐protein, we observed by microscopic examination transfection efficiencies between 70–100%. Three different recombinant proteins were synthesized within a timeframe of 7 days from time of transfection to harvest. The first, a human recombinant IgG1‐type antibody, was secreted into the supernatant of the cell culture and achieved a final concentration of >20 mg/L. An E. coli‐derived DNA‐binding protein remained intracellular, as expected, but accumulated to such a concentration that the lysate of cells, taken up into the entire culture volume, gave a concentration of 18 mg/L. The third protein, a transmembrane receptor, was expressed at 3–6 × 106 molecules/cell. © 2001 John Wiley & Sons, Inc. Biotechnol Bioeng 75: 197–203, 2001.
Validation of a high‐performance liquid chromatographic method for the enantiospecific quantitation of zopiclone in plasma
Authors: Gebauer Markus G. , Alderman Christopher P.
Published: August 2002

Zopiclone is a hypnosedative with clinical effects similar to benzodiazepines but thought to have less potential for rebound insomnia and withdrawal effects. Zopiclone is administered as a racemic mixture, and an enantiospecific method of analysis of zopiclone in plasma is desirable in the study of pharmacokinetic drug interactions. We report a modification of an HPLC method reported by Foster et al. using a closely related structural analogue of zopiclone as internal standard. Zopiclone was detected at 306 nm and linear calibration curves were constructed in the range of 1.0–250 ng/mL for each enantiomer. The % CV at 2.5 ng/mL was 12.0% for (−)‐zopiclone and 14.3% for (+)‐zopiclone, and the limit of quantification of each enantiomer was 2.5 ng/mL. At higher concentrations, the coefficient of variation was less than 10%. The nominal concentration of quality control samples was predicted with an accuracy within a range of ±11.6%. The method was used in the analysis of plasma obtained from psychiatric patients. One sample obtained following a non‐fatal overdose with zopiclone contained the metabolites (−)‐N‐oxide zopiclone and both enantiomers of desmethyl zopiclone. The metabolite enantiomers were resolved on the column with retention times similar to zopiclone. The N‐oxide metabolite co‐eluted with internal standard. Copyright © 2002 John Wiley & Sons, Ltd.
Environmental management of transnational corporations in India—are TNCs creating islands of environmental excellence in a sea of dirt?
Authors: Ruud Audun
Published: August 2002

This paper discusses how and to what extent local environmental practices at affiliated units of transnational corporations (TNCs) are influenced by TNC headquarters (HQ). The study focuses on intra‐firm dynamics of what is termed ‘cross‐border environmental management’ of TNCs. The study documents that the environmental management of TNC‐affiliated units in India are strongly influenced by HQ's environmental policies and standards. However, it is found that there are deviations in local practices from intentions and policy commitments stated at HQ. This can be particularly attributed to local economic and political factors. Cross‐border environmental management is making a difference. However it is limited to affiliated TNC units and few additional external environmental impacts are documented. Copyright © 2002 John Wiley & Sons, Ltd. and ERP Environment
Anatomic relation between the nuchal ligament (ligamentum nuchae) and the spinal dura mater in the craniocervical region
Authors: Dean N.A. , Mitchell B.S.
Published: August 2002

There are inconsistencies between the descriptions of the physical connections between the spinal cervical dura and the surrounding tissues. This study was undertaken to clarify the relationship between the spinal dura, the nuchal ligament and the suboccipital muscles. Dissections were performed on embalmed cadavers: in nine the relevant structures were removed en bloc, whereas in one a sagittal section was prepared. In all specimens it was possible to demonstrate continuity in the midline between the nuchal ligament and the posterior spinal dura at the atlanto‐occipital and atlanto‐axial intervals. No such attachments were found caudal to the arch of the axis. In addition, there was a connective tissue bridge between the deep aspect of the rectus capitis posterior minor muscle to the transverse fibers of the posterior atlanto‐occipital membrane that extended laterally to blend with the perivascular tissue of the vertebral arteries. The present study is, we believe, the first to describe continuity between the nuchal ligament and the dura at the atlanto‐occipital interspace, and confirms previous descriptions of similar connections at the atlanto‐axial level. Knowledge of the exact attachments of the dura may contribute to understanding the biomechanics of the cervical spine and of the possible etiology of some types of cervicogenic headaches. Clin. Anat. 15:182–185, 2002. © 2002 Wiley‐Liss, Inc.
Factors affecting children's disclosure of secrets in an investigatory interview
Authors: Hartwig Janine , Wilson J. Clare
Published: August 2002

The present study aimed to identify how children's understanding of the investigatory interview process influenced their disclosure of secrets. Forty‐four 6‐year‐ and 7‐year‐old children participated in the study. Four short stories were read to each child. The stories were about young children who witnessed an event (e.g. witnessing a person stealing a pizza) and were asked to keep the witnessed event secret, then they were interviewed by an authority figure (e.g. security guard) and asked repeatedly about what they had witnessed. After each story, a series of questions were asked that related to factors that might aid the disclosure of the secret. Three main factors were investigated: children's perception of the interviewer's knowledge of the incident; children's perception of the purpose of the interview; and children's understanding of the purpose of asking repeat questions. Results indicated that a child's perception of how much an interviewer appeared to know about a secret, and whether the child was repeatedly asked about the secret do influence a child's disclosure of a secret. However, there was no effect for the purpose of the interview. Copyright © 2002 John Wiley & Sons, Ltd.
The advanced technology program: A Welfare‐to‐Work success story
Authors: Pagenette Karen , Kozell Cheryl
Published: August 2002

This chapter describes the program, administrative implications, and lessons learned from the Advanced Technology Program, a welfare‐to‐work program developed by Oakland Community College in Michigan, over the past six years.
Stent implantation for long‐segment coarctation of aorta in infant with facial and mediastinal hemangioma
Authors: Matitiau Abraham , Birk Einat , Schonfeld Tommy , Zalzstein Eli , Berant Michael , Bruckheimer Elchanan
Published: August 2002

We report a case of an infant with an extensive hemangioma encompassing the thoracic aorta, associated with complex coarctation. Surgical approach was abandoned for fear of bleeding. The complexity of the coarctation made it unsuitable for balloon dilation. We implanted a stent with significant angiographic improvement and resolution of systemic hypertension. Cathet Cardiovasc Intervent 2002;55:510–512. © 2002 Wiley‐Liss, Inc.
A comparative study of light transmission aggregometry and automated bedside platelet function assays in patients undergoing percutaneous coronary intervention and receiving abciximab, eptifibatide, or tirofiban
Authors: Simon Daniel I. , Liu Christopher B. , Ganz Peter , Kirshenbaum James M. , Piana Robert N. , Rogers MD Campbell , Selwyn MD Andrew P. , Popma Jeffrey J.
Published: August 2002

Platelet inhibition is central to the efficacy of glycoprotein (GP) IIb‐IIIa antagonist therapy, but is not routinely measured during percutaneous coronary intervention (PCI). Data directly comparing the antiplatelet effects of these agents are also limited. Therefore, we compared ex vivo platelet function by standard light transmission aggregometry (LTA) and two automated bedside platelet function assays in 36 patients undergoing PCI with GP IIb‐IIIa inhibitors. At baseline and 10 min following clinically recommended bolus and infusion of abciximab (0.25 mg/kg, 0.125 μg/kg/min), eptifibatide (180 μg/kg, 2 μg/kg/min), or tirofiban (10 μg/kg, 0.1 μg/kg/min), we measured 20 μM ADP‐ and 1.9 mg/mL collagen‐induced platelet aggregation using LTA. Platelet function was also assessed using the bedside Accumetrics Ultegra‐Rapid Platelet Function Assay (RPFA) and the Xylum Clot Signature Analyzer (CSA). The degree of platelet inhibition, as assessed by LTA, varied significantly between the clinically recommended doses of these GP IIb‐IIIa antagonists. RPFA measurements agreed closely with LTA for abciximab, but tended to overestimate the degree of platelet inhibition for small molecules. CSA demonstrated profoundly inhibited shear‐induced platelet function, but lacked sensitivity to discriminate between agents. These findings may have implications for the results of trials comparing the efficacy of these agents in patients undergoing PCI. Cathet Cardiovasc Intervent 2001;52:425–432. © 2001 Wiley‐Liss, Inc.
Reproducibility of intravascular ultrasound and intracoronary Doppler measurements
Authors: Gaster Anne Louise , Korsholm Lars , Thayssen> Per , Pedersen, Knud E. , Haghfelt Torben H.
Published: August 2002

The aim of this study was to evaluate the reproducibility of intravascular ultrasound (IVUS) and intracoronary (IC) Doppler flow velocity measurements. The use of IVUS and IC Doppler has been suggested as a means for percutaneous coronary intervention (PCI) guidance in a series of studies. This would require an acceptable level of accuracy and reproducibility of these two methods for lesion evaluation. In this study, the main focus was on the issue of reproducibility. One hundred and eight patients referred for PCI entered into the study. Inter‐ and intraobserver variability was measured. Catheter difference was assessed. On‐line and off‐line measurements were compared. MUSIC criteria were assessed off‐line, twice. Calculated and measured diameters were compared. After having obtained initial IC Doppler measurements, the Doppler wire was immediately withdrawn and repositioned for reacquisition of Doppler measurements. IVUS measurements are reproducible and reliable off‐line and, to a slightly lesser degree, on‐line. Area measurements should be performed more than once and the mean used for vessel description. Lumen diameters should be calculated from the mean of the area measurements. A measuring technique consensus should be reached and adhered to. CFR measurements can be used to determine reduced vs. normal flow reserve. In this study, it was found that proximal to distal velocity ratio and diastolic to systolic velocity ratio variability made these parameters unsuitable for PCI guidance. Cathet Cardiovasc Intervent 2001;53:449–458. © 2001 Wiley‐Liss, Inc.
Healing response to the clamshell device for closure of intracardiac defects in humans
Authors: Kreutzer Jacqueline , Ryan Carol A. , Gauvreau Kimberlee , Van Praagh Richard , Anderson James M. , Jenkins Kathy J.
Published: August 2002

The healing response to intracardiac devices in humans is largely unknown. During regulatory trials using the Clamshell device in over 800 patients, attempts were made to perform histopathological evaluation of all explanted devices. We reviewed all those with complete histopathological examination (n = 12) from Fontan baffles (n = 4), ventricular septal defects (n = 2), and atrial septal defects (ASD; n = 6), explanted at 2.7 months to 3.6 years (median, 1.6 years), at autopsy (n = 1) or surgery (n = 11), performed for residual defects (n = 5), atrial masses (n = 3), or Fontan revision (n = 3). All but one were nearly (n = 3) or completely (n = 8) covered by pseudointima, composed of fibroelastic tissue, predominantly collagen, with focal foreign body reaction in contact with fabric, without acute inflammation or infection. Atrial masses of granulation tissue were present in three cases (ASD), opposite to protruding fractured arms. No associations were identified between coverage and closure status, position, arm fractures, or implant period. In conclusion, the healing response to transcatheter Clamshell implantation in humans is characterized by a relatively rapid development of a nonthrombotic pseudointima composed of fibroelastic tissue with minimal foreign body reaction. Cathet Cardiovasc Intervent 2001;54:101–111. © 2001 Wiley‐Liss, Inc.
Optimal treatment of nonaorto ostial coronary lesions in large vessels: Acute and long‐term results
Authors: Tan Ren P. , Kini Annapoorna , Shalouh Eli , Marmur Jonathan D. , Sharma Samin K.
Published: August 2002

Percutaneous interventions of nonaorto ostial coronary lesions are usually complex, often requiring a combined approach of debulking and stenting, insertion of multiple guidewires and long procedure duration. Debulking with atherectomy device preserves side‐branch patency by reducing plaque shift while coronary stenting minimizes vessel recoil and restenosis. We retrospectively evaluated the acute and long‐term results of rotational atherectomy (group R, n = 94), coronary stenting (group S, n = 39), and combination of rotational atherectomy and stenting (group R‐S, n = 59) in a total of 192 patients with nonaorto ostial lesions. The number of patients with diabetes mellitus and rest angina was significantly higher in groups S and R‐S. Clinical success rates were high and procedural complication rates were low and comparable in all three groups. Despite the similar reference vessel size and preprocedure minimal lumen diameter (MLD), postprocedure MLD showed a trend toward larger lumen in groups S (3.15 ± 0.18 mm) and R‐S (3.21 ± 0.16 mm). Group S had significantly higher incidence of side‐branch narrowing (30.7%), requiring intervention (15.4%). At long‐term follow‐up (mean of 9 ± 4 months), target lesion revascularization rate was significantly lower in groups R‐S (11.9%) and S (15.4%) compared to group R (28.9%; P = 0.02). Our nonrandomized data suggest that stenting with or without rotational atherectomy provides the best long‐term approach for the interventional treatment of nonaorto ostial coronary lesions. The clinical benefit and cost effectiveness of performing rotational atherectomy before stent implantation to reduce the incidence of side‐branch closure requires further study. Cathet Cardiovasc Intervent 2001;54:283–288. © 2001 Wiley‐Liss, Inc.
Measurement of radial artery spasm using an automatic pullback device
Authors: Kiemeneij Ferdinand , Vajifdar Bhavesh U. , Eccleshall Simon C. , Laarman GertJan , Slagboom Ton , van der Wieken Ron
Published: August 2002

Current evaluation of radial artery spasm (RAS), a frequent finding during the transradial approach for coronary angiography and angioplasty (TRA), is subjective. A quantitative measure of RAS will help in evaluation and comparison of management strategies. The objectives of the study were to assess the feasibility and safety of using an automatic pullback device (APD) for removal of transradial introducer sheaths and to establish a parameter to quantify RAS. In 50 consecutive transradial procedures, the APD was used to measure the force required for sheath removal. The mean maximal pullback force (MPF) was 0.53 ± 0.52 kg (range, 0.1–3.0 kg). In 48 (96%) cases, the MPF was reached within the first 5 sec of pullback. All patients with clinical RAS (n = 4) had an MPF greater than 1.0 kg, while the remaining had an MPF less than 1.0 kg. All patients with severe pain during sheath removal (n = 3) had an MPF greater than 1.0 kg, while no patient with an MPF less than 1.0 kg had severe pain. It is feasible and safe to remove transradial introducer sheaths using the APD. The MPF is achieved within the first 5 sec of pullback and is a reliable parameter to quantify RAS. An MPF more than 1.0 kg correlates with clinical RAS and is associated with severe pain during sheath removal. Cathet Cardiovasc Intervent 2001;54:437–441. © 2001 Wiley‐Liss, Inc.
Centrosome and spindle pole microtubules are main targets of a fluorescent taxoid inducing cell death
Authors: Abal Miguel , Souto André A. , Amat‐Guerri Francisco , Acuña A. Ulises , Andreu Jose M. , Barasoain Isabel
Published: August 2002

Microtubules offer a very large local concentration of binding sites for cytotoxic taxoids or for hypothetical endogenous regulators. Several compounds from diverse sources stabilize microtubules and arrest cell division similarly to the antitumour drug Taxol. We have investigated the subcellular location of the Taxol binding sites, employing a fluorescent taxoid (FLUTAX) that reversibly interacts with the Taxol binding sites of microtubules and induces cellular effects similar to Taxol. The specific binding of FLUTAX to a fraction of the available cellular binding sites effectively inhibits division of cultured human tumour cells at G2/M, and triggers apoptotic death. The loci of reversible binding, directly imaged in intact U937 cells treated with cytotoxic doses of fluorescent taxoid are the centrosomes, with a few associated microtubules in interphase cells, and the spindle pole microtubules in mitotic cells, instead of uniformly labelling the microtubule cytoskeleton. Cytoskeletal lesions induced and visualized with FLUTAX consist of microtubule bundles and abnormal mitoses, including monopolar spindles and highly fluorescent multipolar spindles. The multiple asters and monopolar spindles mark arrested U937 leukaemia and OVCAR‐3 ovarian carcinoma cells on their path to apoptosis (as well as K562, HeLa, and MCF‐7 cells). Depending on the FLUTAX treatment, OVCAR‐3 cells died from abnormal mitosis or from a subsequent interphase with double chromatin content and lobulated nuclei (micronuclei), indicating impairment of centrosome separation. Fragmented centrosomes could be observed in FLUTAX‐treated non‐transformed 3T3.A31 cells, which developed micronuclei but were resistant to apoptosis. These results strongly suggest that centrosomal impairment by taxoid binding during interphase, in addition to the suppression of the kinetochore microtubule dynamics in the mitotic spindle, is a primary cause of cell cycle de‐regulation and cell death. Cell Motil. Cytoskeleton 49:1–15, 2001. © 2001 Wiley‐Liss, Inc.
De novo formation of cytokeratin filament networks originates from the cell cortex in A‐431 cells
Authors: Windoffer Reinhard , Leube Rudolf E.
Published: August 2002

Of the three major cytoskeletal filament systems, the intermediate filaments are the least understood. Since they differ fundamentally from the actin‐ and microtubule‐based networks by their lack of polarity, it has remained a mystery how and where these principally endless filaments are formed. Using a recently established epithelial cell system in which fluorescently labeled intermediate filaments of the cytokeratin type can be monitored in living cells, we address these issues. By multidimensional time‐lapse fluorescence microscopy, we examine de novo intermediate filament network formation from non‐filamentous material at the end of mitosis and show that it mirrors disassembly. It is demonstrated that filament formation is initiated from the cell cortex without focal preference after cytokinesis. Furthermore, it is shown that this process is dependent on energy, on the integrity of the actin filament network and the microtubule system, and that it can be inhibited by the tyrosine phosphatase inhibitor pervanadate. Based on these observations, a two‐step working model is proposed involving (1) interactions within the planar cortical layer acting as an organizing center forming a two‐dimensional network and (2) subsequent radial dynamics facilitating the formation of a mature three‐dimensional network. Cell Motil. Cytoskeleton 50:33–44, 2001. © 2001 Wiley‐Liss, Inc.
Spin‐1 nuclear quadrupole resonance theory with comparisons to nuclear magnetic resonance
Authors: Lee Y.K.
Published: August 2002

The theory of pure spin‐1 nuclear quadrupole resonance (NQR) excitation and detection is presented with comparisons to nuclear magnetic resonance (NMR). Even though the dominating Hamiltonians in pure NQR and NMR have different symmetry properties, in both single resonant frequency spin‐1 NQR and spin‐½ NMR the spin dynamics are isolated to two resonant energy levels, leading to close parallels in the calculations of their respective signals. The theory of spin‐1 NQR is presented in terms of matrix calculations and fictitious spin‐½ operators, and the analysis focuses on the similarities and differences of the spin‐1 NQR and spin‐½ NMR physical interpretations and rotational symmetry properties. © 2002 Wiley Periodicals, Inc. Concepts in Magn Reson 14, 155–171, 2002.
Expression of CD40 ligand (CD154) in B and T lymphocytes of Hodgkin disease
Authors: Clodi Katharina , Asgari Zahra , Younes Mamoun , Palmer J. Lynn , Cabanillas Fernando , Carbone Antonino , Andreeff Michael , Younes Anas
Published: August 2002

BACKGROUND

The malignant Hodgkin and Reed–Sternberg (H/RS) cells of Hodgkin disease (HD) express CD30 and CD40 receptors that can activate nuclear factor kappa B and transduce survival signals. The authors have reported previously that the B lymphocytes of HD express CD30 ligand (CD30L, CD153). Furthermore, they and others have reported previously that the CD40L survival pathway is augmented in patients with B‐cell malignancies, as CD40L was constitutively expressed by the malignant B cells and infiltrating T cells, and sera from those patients contained elevated levels of soluble CD40L. In this study, the authors investigated the hypothesis that the survival of H/RS cells was similarly promoted by an augmented CD40L signals in HD patients.

METHODS

The expression of CD40L on lymphocyte subsets of patients with classic HD was determined by two‐color fluorescent‐activated cell sorter analysis. Serum soluble CD40L levels were determined by enzyme linked immunosorbent assay.

RESULTS

CD40L was constitutively expressed on both the T and B cells of HD patients but was more prominently expressed on the B lymphocytes. Soluble CD40L was detected in the serum of 17 of 37 patients (45%) and was higher than 1 ng/mL in 4 patients (10%). Both interleukin (IL)‐4 and IL‐10, which are known to be secreted by H/RS cells and surrounding T cells, up‐regulated CD40L expression on normal B cells.

CONCLUSIONS

Thus, the expression of CD40L and CD30L on the B cells of HD patients suggests that B lymphocytes may play a role in the regulation of H/RS cell growth in vivo. Depriving H/RS cells from CD30L and CD40L survival signals by eliminating B cells from HD lesions may be of therapeutic value. Cancer 2002;94:1–5. © 2002 American Cancer Society.
A phase II study of neoadjuvant biochemotherapy for stage III melanoma
Authors: Gibbs Peter , Anderson Clay , Pearlman Nathan , LaClaire Stacy , Becker Maude , Gatlin Kristi , O'Driscoll Martin , Stephens Janet , Gonzalez Rene
Published: August 2002

BACKGROUND

Phase II studies of biochemotherapy (combining interleukin‐2, interferon‐α, and multiagent chemotherapy) have reported high response rates and a significant number of durable complete responses in patients with metastatic melanoma.

METHODS

A pilot Phase II study was performed to explore the safety and activity of neoadjuvant biochemotherapy in patients with Stage III melanoma. Forty‐eight patients were enrolled between April 1996 and May 1999. The median age of the patients was 46 years (range, 19–70 years). Two cycles of biochemotherapy were administered prior to and after complete lymph node dissection. Each cycle was comprised of cisplatin, 20 mg/m2 intravenously (i.v.), on Days 1–4; vinblastine, 1.6 mg/m2 i.v., on Days 1–4; dacarbazine, 800 mg/m2 i.v., on Day 1; interleukin‐2, 9 × 106 IU/m2/day i.v. over 24 hours, on Days 1–4; and interferon‐α, 5 × 106 IU/m2/day subcutaneously, on Days 1–5, every 3 weeks. Twelve patients did not have measurable disease. All patients were evaluable for toxicity and survival.

RESULTS

Clinical responses were observed in 14 of 36 patients (38.9%) with measurable disease, including 13 partial responses (36.1%) and 1 complete response (2.8%). Complete pathologic responses were noted in 4 patients (11.1%). Toxicity, although severe, was manageable and typically short‐lived. There were no treatment‐related deaths reported. At a median follow‐up of 31 months, 38 of the 48 patients (79.2%) were alive and 31 patients (64.6%) remained free of disease progression.

CONCLUSIONS

Neoadjuvant biochemotherapy appears to have promising activity in patients with Stage III melanoma. A larger multicenter study currently is underway to explore this approach further. Cancer 2002;94:470–6. © 2002 American Cancer Society.
Molecular markers of prognosis in astrocytic tumors
Authors: Rasheed Ahmed , Herndon James E. , Stenzel Timothy T. , Raetz Jacqueline G. M. , Kendelhardt Jason , Friedman Henry S. , Friedman Allan H. , Bigner Darell D. , Bigner Sandra H. , McLendon Roger E.
Published: August 2002

BACKGROUND

Astrocytoma is a primary brain tumor that affects 20,000 Americans each year. To date, only age and histologic grade stand out as independent predictors of survival. There is now increased interest in the use of molecular markers as objective standards against which to establish diagnosis and grade.

METHODS

The study evaluated human glioma tumor suppressor genes and associated loci in fresh snap‐frozen gliomas from 63 males and 37 females, with a median age of 42 years, including 19 low‐grade astrocytomas. The tumor samples were selected so that about equal numbers of glioblastomas from younger and older patients were represented in the series. Methods for suppressor gene and genetic loci evaluation included loss of heterozygosity (LOH) analysis, multiplex polymerase chain reaction analysis, and gene sequencing.

RESULTS

Low‐grade astrocytomas had the least number of molecular abnormalities. LOH on 9p and/or CDKN2A deletion occurred more often in glioblastomas (P < 0.001), LOH on 17p/TP53 mutations occurred more frequently in anaplastic astrocytomas (AAs; P = 0.112), and LOH on 10q/PTEN mutation frequency was similar in glioblastomas and AAs (P < 0.001). Poorer survival was associated significantly with the occurrence of either deletion of p16 (P = 0.031), LOH on 9p (P = 0.016), or LOH on 10q (P = 0.0007). The absence of LOH on 17p and the presence of PTEN mutation were associated marginally with survival. Even though TP53 mutations were more frequent among younger patients with glioblastoma, they had no statistically significant effect on survival after adjustment for age (P = 0.62). In all multivariate models, age and grade were the only significant predictors of survival or were nearly significant predictors of survival.

CONCLUSIONS

The results suggest that LOH on 9p and p16 deletions may prove to be objective standards for the diagnosis of patients with high‐grade gliomas, although the absence of these abnormalities is nonprognostic. Cancer 2002;94:2688–97. © 2002 American Cancer Society.

DOI 10.1002/cncr.10544
ErbB2 overexpression correlates with increased expression of vascular endothelial growth factors A, C, and D in human breast carcinoma
Authors: Yang Wentao , Klos Kristine , Yang Ying , Smith Terry L. , Shi Daren , Yu Dihua
Published: August 2002

BACKGROUND

The angiogenic factor vascular endothelial growth factor (VEGF)‐A plays an important role in breast cancer progression. However, the involvement of VEGF‐C and VEGF‐D, two newer members of the VEGF family, in breast carcinoma and their relationship with clinicopathologic parameters have not been clearly demonstrated.

METHODS

In this study, the expression levels of VEGF‐A, VEGF‐C, and VEGF‐D protein in 107 breast carcinoma cases and 22 nonmalignant breast tissue samples were examined by immunohistochemistry and quantitated by image analysis.

RESULTS

Higher expression of VEGF‐C and VEGF‐D was found in breast carcinomas than in nonmalignant breast tissue samples. Moreover, expression of VEGF‐A, VEGF‐C, and VEGF‐D was significantly and positively correlated with ErbB2 expression. High levels of VEGF‐A expression were associated with shorter disease‐free survival (DFS). Patients with tumors expressing high levels of VEGF‐C or VEGF‐D showed a notable trend for worse DFS, however, it was not statistically significant. The combination of VEGF‐A and VEGF‐C status predicted survival better than either marker alone.

CONCLUSIONS

Our study suggests that expression of the angiogenic and lymphangiogenic factors (i.e., VEGFs) might be regulated at least in part by ErbB2. In addition, the combination of VEGF‐A and VEGF‐C status may better predict prognosis of patients with breast carcinoma than VEGF‐A alone. Cancer 2002;94:2855–61. © 2002 American Cancer Society.

DOI 10.1002/cncr.10553
Neuronal development in larval chiton Ischnochiton hakodadensis (Mollusca: Polyplacophora)
Authors: Voronezhskaya Elena E. , Tyurin Sergei A. , Nezlin Leonid P.
Published: August 2002

Chitons are the most primitive molluscs and, thus, a matter of considerable interest for understanding both basic principles of molluscan neurogenesis and phylogeny. The development of the nervous system in trochophores of the chiton Ischnochiton hakodadensis from hatching to metamorphosis is described in detail by using confocal laser scanning microscopy and antibodies raised against serotonin, FMRFamide, and acetylated α‐tubulin. The earliest nervous elements detected were peripheral neurons located in the frontal hemisphere of posthatching trochophores and projecting into the apical organ. Among them, two pairs of unique large lateral cells appear to pioneer the pathways of developing adult nervous system. Chitons possess an apical organ that contains the largest number of neurons among all molluscan larvae investigated so far. Besides, many pretrochal neurons are situated outside the apical organ. The prototroch is not innervated by larval neurons. The first neurons of the developing adult central nervous system (CNS) appear later in the cerebral ganglion and pedal cords. None of the neurons of the larval nervous system are retained in the adult CNS. They cease to express their transmitter content and disintegrate after settlement. Although the adult CNS of chitons resembles that of polychaetes, their general scenario of neuronal development resembles that of advanced molluscs and differs from annelids. Thus, our data demonstrate the conservative pattern of molluscan neurogenesis and suggest independent origin of molluscan and annelid trochophores. J. Comp. Neurol. 444:25–38, 2002. © 2002 Wiley‐Liss, Inc.
Central organization of the electrosensory system in the paddlefish (Polyodon spathula)
Authors: Hofmann Michael H. , Wojtenek Winfried , Wilkens Lon A.
Published: August 2002

The central connections of the electrosensory system were studied in the paddlefish Polyodon spathula by injecting biotinylated dextran amines into the dorsal octavolateral nucleus (DON), the cerebellum, and the mesencephalic tectum. The sole target of primary electrosensory fibers is the ipsilateral dorsal octavolateral nucleus. The principal neurons ascending from this nucleus project to the torus semicircularis, the lateral mesencephalic nucleus, and the mesencephalic tectum. The mesencephalic tectum projects back to the nucleus preeminentialis, which, in turn, projects to the cerebellar auricles and to the DON. The auricles are the main source of parallel fibers in the cerebellar crest ventral to the DON. The DON also receives input from the contralateral DON. These descending feedback loops are very similar to those of other electrosensory fishes. However, the paddlefish is unique in having three mesencephalic targets of electrosensory information. It is the only bony fish known to have extensive projections directly to the mesencephalic tectum and to a lateral mesencephalic nucleus in addition to the torus semicircularis. J. Comp. Neurol. 446:25–36, 2002. © 2002 Wiley‐Liss, Inc.
Callosal connections correlate preferentially with ipsilateral cortical domains in cat areas 17 and 18, and with contralateral domains in the 17/18 transition zone
Authors: Olavarria Jaime F.
Published: August 2002

Previous studies have shown that the distribution of callosal connections in the 17/18 callosal zone of the cat is patchy at a small scale, but the mechanisms that determine this periodic pattern remain unclear. The present study investigated this issue by correlating the distribution of retrogradely labeled callosal cells with the underlying patterns of ocular dominance columns (ODCs) revealed transneuronally after intraocular injections of wheat germ agglutinin‐horseradish peroxidase. The density of labeled callosal cells was found to vary significantly between adjacent territories dominated by different eyes, indicating that the distribution of callosal cells is significantly biased toward domains that are eye specific. Moreover, callosal connections relate to the pattern of ODCs in a rather unique way: callosal cells correlate preferentially with contralateral ODCs within the 17/18 transition zone (TZ), and with ipsilateral ODCs in regions of areas 17 and 18 located outside the TZ. Similar results were obtained in cats raised with strabismus, indicating that the overlap between right and left ODCs present in normal cats does not influence the correlation between callosal neurons and ODCs. The results are consistent with the hypothesis that callosal linkages are stabilized during development by interhemispheric correlated activity driven by bilateral projections from temporal retina. It is proposed that developmental constraints imposed by both this retinally driven mechanism and the pattern of ODCs are likely to determine not only the association of callosal clusters with specific sets of ODCs, but also important aspects of the functional characteristics of the callosal pathway in cat striate cortex. J. Comp. Neurol. 433:441–457, 2001. © 2001 Wiley‐Liss, Inc.
Catecholamine‐containing cells in the central nervous system and periphery of Aplysia californica
Authors: Croll Roger P.
Published: August 2002

Previous studies have suggested the presence of numerous catecholamine‐containing cells in both the central ganglia and peripheral tissues of Aplysia, but they often offered conflicting or incomplete accounts of numbers, locations, and morphologies. The current study combines aldehyde‐induced histofluorescence and tyrosine hydroxylase‐like immunoreactivity together with confocal microscopy to provide details of these cells. Approximately 35–50 neurones in the cerebral ganglia, 4–8 neurones in the pedal ganglia, 5 neurones in the buccal ganglia, and numerous small fibres in various nerve trunks exhibited both immunoreactivity and aldehyde‐induced fluorescence. Approximately 20 cells in the pedal ganglia and 4 cells in the buccal ganglia exhibited only immunoreactivity whereas 15–20 neurons in the cerebral ganglia exhibited only aldehyde‐induced fluorescence. No somata in the pleural or abdominal ganglia exhibited aldehyde‐induced fluorescence or immunoreactivity. Both aldehyde‐induced histofluorescence and immunoreactivity also labelled what appeared to be two classes of catecholamine‐containing cells in the gill, siphon, oesophagus, rhinophore, tentacle, and reproductive organs. The more numerous, but smaller cells had subepithelial somata and processes penetrating the overlying body wall, thus suggesting a sensory function. Another class of neurones had larger somata, often located more deeply within the tissue, and occasionally appeared to be multipolar. Processes from these various peripheral cells appeared to comprise the major component of afferent fibres and to form an extensive peripheral plexus, often associated with various muscles. The morphologies of the peripheral cells thus suggest involvement in both local and centrally mediated reflexes and responses, but additional studies must test such hypothesised functions and determine the sensory modalities that the cells mediate. J. Comp. Neurol. 441:91–105, 2001. © 2001 Wiley‐Liss, Inc.
Three‐dimensional reconstruction of scleral cold thermoreceptors of the cat eye
Authors: Heppelmann Bernd , Gallar Juana , Trost Brigitte , Schmidt Robert F. , Belmonte Carlos
Published: August 2002

Sensory endings that respond to local cooling were identified electrophysiologically in the cat's sclera. Functionally identified scleral thermal fibers were then used to analyze the structural characteristics of cold receptor endings. Four Aδ units sensitive to controlled cooling of their scleral receptive fields were recorded. The receptive areas were mapped, demarcated with pins and examined electron microscopically using extensive three‐dimensional reconstructions. The supporting tissue within the receptive areas of cold units consisted of dense collageneous tissue with a small number of blood vessels that were either veins or capillaries. Aδ nerve fibers were found within these tissue blocks presumably corresponding with cold sensitive fibers. Small nerves and single nerve fibers devoid of a perineurium were found in all parts of the tissue, only occasionally passing a blood vessel. The terminal portions showed axonal swellings all along the unmyelinated segment filled with mitochondria, glycogen particles, and some vesicles. About 30% of the terminal axonal membrane is not covered by Schwann cells. In the unmyelinated distal portion, the mitochondrial content ranged from 0.012 to 0.038 μm3 mitochondrial volume per μm2 nerve fiber membrane. In comparison with sensory endings in the cat's knee joint, cold receptors in the cat sclera showed many similarities in their three‐dimensional structure with polymodal nociceptor endings of the knee joint but contain less mitochondria. This suggests that cold sensory endings do not require specialized cellular processes for the transduction of cold stimuli, as is the case for multimodal transduction and sensitization in the terminal portion of polymodal nociceptors. J. Comp. Neurol. 441:148–154, 2001. © 2001 Wiley‐Liss, Inc.
CD45‐assisted PanLeucogating for accurate, cost‐effective dual‐platform CD4+ T‐cell enumeration
Authors: Glencross Deborah , Scott Lesley E. , Jani Ilesh V. , Barnett David , Janossy George
Published: August 2002

BACKGROUND

North American and European guidelines for dual‐platform (DP) flow cytometry recommend absolute CD4 T‐cell counts to be calculated from two parameters: the absolute lymphocyte counts obtained on a hematology analyzer and the percentages of CD4+ cells among lymphocytes (CD4%/lympho) obtained by flow cytometry. Nevertheless, the identification of lymphocytes is error‐prone: a poor match between these common denominators in the two systems is the main source of inaccuracy. In contrast, total leucocyte counts (white cell counts [WCC]) and CD4% among the gated CD45+ leucocytes (CD4%/leuco) can be determined with greater accuracy. Methods: We introduced “PanLeucogating,” i.e., we used total leucocytes as the common denominator for improving the precision of DP absolute CD4 counting. Correlations and Bland‐Altman tests were used for statistical analysis. Results: First, 22 stabilized blood product samples were provided by U.K. National External Quality Assessment Scheme (NEQAS) and a higher accuracy and precision of CD4 counts were documented using PanLeucogating compared with lymphocyte gating. Next, 183 fresh and 112 fixed (TransFix) whole blood samples were used to compare DP methods and single‐platform (SP) methodology, including both volumetric and bead‐based techniques. A particularly high correlation and comparable precision of absolute CD4 counts were observed between the SP volumetric method and DP PanLeucogating (R2 = 0.990; bias 6 ± SD 17%). The SP volumetric method showed lower levels of agreement with the DP lymphocyte gating (R2 = 0.758; bias 14 ± SD 51%) and with the SP bead‐based method (R2 = 0.923; bias 4 ±SD 31%). Conclusions: These observations show that DP leucocyte counts (WCC) should replace lymphocyte counts as the “common denominator” although CD4%/lympho values can, as an extra step, be also provided readily if requested. When coupled with quality control for WCC on hematology analyzers, the DP method with CD45 PanLeucogating represents a robust CD4 T‐cell assay that is as accurate as the SP volumetric technique. This DP method uses only two, CD45 and CD4, antibody reagents and can be run on any pair of hematological analyzer plus flow cytometer. Cytometry (Clin. Cytometry) 50:69–77, 2002. © 2002 Wiley‐Liss, Inc.
Selection of lymphocyte gating protocol has an impact on the level of reliability of T‐cell subsets in aging specimens
Authors: Bergeron M. , Nicholson J.K.A. , Phaneuf S. , Ding T. , Soucy N. , Badley A.D. , Hawley Foss N.C. , Mandy F.
Published: August 2002

BACKGROUND

In the past decade, human immunodeficiency virus (HIV) lymphocyte immunophenotyping has evolved significantly. New fluorochromes, new multicolor reagents, enhanced instruments, and the capacity to provide absolute cell counts using the single‐platform technique have all contributed to the reliability of T‐cell subset measurements. In this study, four gating protocols were evaluated to select the most robust method for T‐cell subset enumeration. Methods: Peripheral blood specimens from 21 HIV+ and 20 HIV− individuals were monitored up to 96 h. Aliquots of specimens were stored at room temperature and analyzed at 6 (baseline), 48, 72, and 96 h. Aliquots were stained with CD45‐fluorescein isothiocyanate (FITC)/CD3PC5/CD4RD1/CD8ECD. Data analysis was performed with all four gating protocols. Results: Only with fresh blood did all protocols provide similar results. From samples that were 48 h old, the choice of gating strategy had a dramatic impact on immunophenotyping results. The largest deviations from baseline values occurred at 96 h and gating protocols that included dual light scatter gates provided the greatest shift of T‐cell subset values over time. The gating protocols that were based exclusively on cell lineage‐specific gates gave the most robust T‐cell values up to 96 h. Conclusion: By selecting the appropriate gating protocol, the temporal integrity of specimens can be extended up to 4 days. Cytometry (Clin. Cytometry) 50:53–61, 2002. © 2002 Wiley‐Liss, Inc.
Malignant pleural effusions due to small‐cell lung carcinoma: A cytologic and immunocytochemical study
Authors: Chhieng David C. , Ko Ellen C. , Yee Herman T. , Shultz Jane J. , Dorvault Christine C. , Eltoum Isam A.
Published: August 2002

Patients with small‐cell lung carcinoma (SCLC) rarely present with pleural effusions. Based on morphology alone, recognition of SCLC in effusion cytology may be challenging because of the resemblance of neoplastic cells to lymphocytes. Immunocytochemistry may be helpful in its diagnosis. The objective of this study was to review the morphology and evaluate the use of immunocytochemistry in diagnosing SCLC in pleural fluids. Patients with SCLC who presented with pleural effusions were identified during a 6‐yr period. The cytology and medical records were reviewed. Formalin‐fixed, paraffin‐embedded cell blocks of fluid specimens were immunostained with neuroendocrine markers (chromogranin A and synatophysin), cytokeratin 20 (CK20), and thyroid transcription factor‐1 (TTF‐1). The latter is a nuclear transcription protein that is expressed in normal respiratory epithelium and also in more than 90% of SCLCs. Of the 256 patients diagnosed with SCLC during the study period, 8 (2.7%) patients (3 females and 4 males, age range from 56–85 yr) also developed pleural effusions. One patient had 2 fluid specimens during the course of their disease, giving a total of 9 specimens. Four specimens had a positive cytologic diagnosis of SCLC, and 2 were initially diagnosed as suspicious for SCLC. The remaining 3 specimens were negative for SCLS. The specimens with a positive or suspicious diagnosis showed single and aggregates of small to medium‐sized single cells with a high nuclear:cytoplasmic (N:C) ratio, round to angulated nuclei, and salt‐and‐pepper chromatin. Nuclear molding was also noted. Five out of 6 (83%) specimens with a positive or suspicious diagnosis of SCLC were positive for both chromogranin A and TTF‐1. Synaptophysin was positive in 3 of 6 (50%) positive or suspicious cases. None of the cases were positive for CK20. All cases with a negative cytologic diagnosis were negative for chromogranin A, synatophysin, CK20, and TTF‐1. In conclusion, patients with SCLC rarely present with pleural effusions. The cytology of SCLC is characteristic. The use of immunocytochemistry, particularly with antibodies to chromogranin A, TTF‐1, and CK 20, aids in the differential diagnosis. Diagn. Cytopathol. 25:356–360, 2001. © 2001 Wiley‐Liss, Inc.
Cervical biopsy/cytology correlation data can be collected prospectively and shared clinically
Authors: Cioc Adina M. , Julius Carmen J. , Proca Daniela M. , Tranovich Virginia L. , Keyhani‐Rofagha Sedigheh
Published: August 2002

Cervical cytology (Cy) and biopsy (Bx) correlation is used by institutions for the evaluation of their cytodiagnostic capabilities as a part of overall laboratory quality improvement (QI). However, the data obtained from correlation are not routinely included in most surgical pathology (SP) reports. Our laboratory's procedure is to include the correlation of the patient's previous (most recent) cytology smear in the surgical pathology report of all/any gynecologic surgical pathology specimens. We reviewed this process for the time period between July 1998–June 1999. Any noncorrelating cases were assigned a correlation review code by the reviewing cytopathologist: major Cy diagnostic error (DE1), minor Cy diagnostic error (DE2), Cy sampling error (Cy SE), or biopsy sampling error (Bx SE). Of 3,486 cases reviewed, 3,229 cases were satisfactory for correlation studies. Concordant results were found in 86.9%. Cy DE1 due to either Cy screening or interpretation errors or both were found in 0.2% (n = 7) of all cases, while Cy DE2 due to the same were found in 1% (n = 32). Bx SE accounted for discrepancies in 6.8% (n = 220) of all cases, while 5.1% (n = 164) of the total cases were discrepancies due to Cy SE. Follow‐up Bx was available in 97.2% (n = 214) of the Bx SE, and showed 16.4% (n = 35) to be major discrepancies and 83.6% (n = 179) to be minor discrepancies. Cervical Cy/Bx correlation is useful for the evaluation of a laboratory's QI. It is also useful for the identification of either Cy or Bx SE. While QI data exist as “internal use only” documents, SE data (as part of the CC (correlation comment) included in SP reports) are vital to a specific/given patient. Bx SE was identified in 6.3% of our patients, indicating a possible need for rebiopsy. This type of QI data may be shared clinically, and may direct the management for maximum diagnostic and patient benefit. Diagn. Cytopathol. 2002;26:49–52. © 2002 Wiley‐Liss, Inc.