bluuebunny/crossref_metadata_2025 huggingface dataset:
A note on 3‐factorizations of K10*
Authors: Khosrovshahi G. B. , Maysoori Ch. , Tayfeh‐Rezaie B.
Published: August 2002

In this note, we enumerate all nonisomorphic 3‐factorizations of K10. © 2001 John Wiley & Sons, Inc. J Combin Designs 9: 379–383, 2001
Possible automatic cell classification of bone marrow aspirate using the CELL‐DYN 4000® automatic blood cell analyzer
Authors: Yamamura Ryousuke , Yamane Takahisa , Hino Masayuki , Ohta Kensuke , Shibata Hisako , Tsuda Izumi , Tatsumi Noriyuki
Published: August 2002

In clinical hematology, the demand for bone marrow aspiration testing is increasing. However, conventional automatic blood cell analyzers cannot completely analyze erythroblasts, and evaluation has mainly been performed by visual examination (the microscopic method). Using the CELL‐DYN 4000® automatic blood cell analyzer (CD4000) (Abbott Laboratories, North Chicago, IL), specific recognition and classification of erythroblasts by DNA staining is possible. In the present study, using bone marrow blood collected from normal subjects and patients with hematological malignancy, we classified cells by the microscopic method and with the CD4000, and compared the results. Good correlations were found for total nucleated cell count (TNCC), neutrophils, lymphocytes, erythroblasts, and the myeloid series to erythroid series (M/E) ratio. It is possible to detect blasts that emerge in patients with hematological malignancy using the blast flag system installed on the CD4000. Since all of the items can be analyzed in about 80 sec with the CD4000, cells in bone marrow aspirates can be classified faster with this apparatus than by the microscopic method. Therefore, analysis of bone marrow aspirates with this apparatus appears to be very useful not only for laboratory testing but also for clinical screening. J. Clin. Lab. Anal. 16:86–90, 2002. © 2002 Wiley‐Liss, Inc.
Commentary: Advances in the laboratory diagnosis of celiac disease
Authors: McPherson Richard A.
Published: August 2002

Anti‐endomysial autoantibodies are very useful in the diagnosis and monitoring of celiac disease (gluten‐sensitive enteropathy) due to their high sensitivity and specificity for that disorder. The recent discovery that the autoantigen responsible for the endomysial pattern is tissue transglutaminase (tTG) has led to the commercial development of automated enzyme immunoassays for quantitation of that autoantibody. These assays are standardized to provide highly accurate and comparable testing between laboratories for anti‐tTG autoantibodies. Celiac disease is a common genetic disease in populations of Europe and the United States. It has a spectrum of expression ranging from silent or mild to severe, with resulting malabsorption that produces multiple‐organ system effects due to malnutrition. Many cases miss the diagnosis because the symptoms are not classic or the clinical syndrome is not severe. Because the treatment of celiac disease (avoidance of wheat products) is so effective and inexpensive and because celiac disease is so common in selective populations, a highly reliable test for its detection such as anti‐tTG should find wide application in clinical practice. J. Clin. Lab. Anal. 15:105–107, 2001. © 2001 Wiley‐Liss, Inc.
IgA antibodies against endomysium and transglutaminase: A comparison of methods
Authors: Jaskowski Troy D. , Schroder Carl , Martins Thomas B. , Litwin Christine M. , Hill Harry R.
Published: August 2002

Recently, the endomysial antigen has been identified as the protein cross‐linking enzyme known as tissue transglutaminase (tTG). Our objective was to compare a novel enzyme immunoassay (EIA) that detects IgA antibody against tTG to two standard IFA methods utilizing thin tissue sections of rat kidney/rat stomach (KS) and distal primate esophagus (PE) as substrates to detect IgA antibody against endomysium (EMA). Sera from 100 patients suspected of having gluten‐sensitive enteropathy (GSE) and 23 sera possessing various antibodies used for EIA cross‐reactivity studies were included. Additional tests, performed routinely in our laboratory, were utilized to further assess sera from patients suspected having GSE. These tests include anti‐gliadin IgA antibody (AGA) and anti‐reticulin IgA antibody (ARA) and are part of the European Society for Pediatric Gastroenterology and Nutrition (ESPGAN) revised criteria for diagnosing GSE. When compared to IFA using KS, the tTG EIA had a sensitivity of 87.5%, was 97.1% specific, and had an overall agreement of 94.0%. When compared to IFA using PE, the tTG EIA had a sensitivity of 92.6%, was 93.2% specific, and had an overall agreement of 93.0%. When the KS IFA was compared to the PE IFA for EMA, the KS IFA had a sensitivity of 96.3%, was 91.8% specific, and had an overall agreement of 93.0%. The majority of sera that were positive for tTG but were negative by IFA (KS, n = 2/PE, n = 5) possessed IgA antibodies against gliadin and/or reticulin. Five of six sera with negative results by PE IFA were positive by the KS IFA and possessed one or more antibodies to tTG and/or gliadin and/or reticulin. We conclude that the tTG EIA compares well to both KS and PE IFAs when detecting IgA antibody against endomysium. We do not recommend the use of PE to detect EMA primarily because of the inconsistencies (i.e., tissue selection, quality, and preparation) and limited availability of commercially prepared PE tissue. J. Clin. Lab. Anal. 15:108–111, 2001. © 2001 Wiley‐Liss, Inc.
Differential distribution of soluble and complexed forms of prostate‐specific antigen in cyst fluids of women with gross cystic breast disease
Authors: Malatesta Manuela , Mannello Ferdinando , Sebastiani Maurizio , Gazzanelli Giancarlo
Published: August 2002

Gross cystic breast disease (GCBD) is the most common benign disease of the human female breast, and patients with GCBD have an increased risk of breast cancer. The aim of this study was to evaluate the distribution inside apocrine cells and in breast cyst fluids aspirated from gross cysts of prostate‐specific antigen (PSA) molecular forms, and to correlate the different intracystic PSA profiles to the subpopulations of gross cysts. Type I cysts showed a median value of 0.71 &mgr;g/L of total PSA and 0.32 g/L of ACT, significantly different to that of Type II cysts (Wilcoxon P < 0.001). Although large excesses of ACT were detected in all samples, BCF samples and apocrine cells from Type I gross cysts contained about 70% of free PSA, compared to the higher amounts of complexed PSA found in Type II gross cysts. We demonstrate that in apocrine/secretive Type I breast gross cysts the serine protease PSA was mainly present in its free form, in contrast to a major proportion of complexed PSA found in flattened/transudative Type II cysts. Our results are consistent with the notion that a prolonged exposure of apocrine breast cells lining the Type I gross cysts to the proteolytic activity of PSA could be involved in the etiopathogenesis of GCBD. J. Clin. Lab. Anal. 15:81–86, 2001. © 2001 Wiley‐Liss, Inc.
An examination of the psychometric structure of the Multidimensional Pain Inventory
Authors: Deisinger Julie A. , Cassisi Jeffrey E. , Lofland Kenneth R. , Cole Patricia , Bruehl Stephen
Published: August 2002

Factor analyses to create the Multidimensional Pain Inventory (MPI) warrant further examination due to small sample size, the use of separate factor analyses for each MPI section, and lack of evidence for the replicability of the factor model. The present study randomly assigned 972 respondents to one of three samples. Study 1 used data from Sample 1 (n = 452) to examine the replicability of the MPI factor structure reported by Kerns, Turk, and Rudy (1985), and to measure the internal consistencies of MPI scales. In Study 2, confirmatory factor analyses using Sample 2 data (n = 267) verified a revised MPI factor structure that was generated in Study 1; cross‐validation was achieved using data from Sample 3 (n = 253). The revised MPI model differed from the original model in its number of scales and pattern of scale loadings. Recommendations for the refinement of the MPI are proposed. © 2001 John Wiley & Sons, Inc. J Clin Psychol 57: 765–783, 2001.
A psychodynamic perspective on resistance in psychotherapy: Vive la résistance
Authors: Messer Stanley B.
Published: August 2002

The term resistance has an overly negative connotation, indicating a recalcitrant, oppositional tendency on the part of psychotherapy clients. This article emphasizes the inevitability and ubiquity of resistance and argues that it should be greeted as a therapist's friend, not as an enemy. It is the way in which clients present themselves to the world in general and to the therapist in particular. Five forms of resistance are presented, including: resistance to the recognition of feelings, fantasies, and motives; resistance to revealing feelings toward the therapist; resistance as a way of demonstrating self‐sufficiency; resistance as clients' reluctance to change their behavior outside the therapy room; and resistance as a function of failure of empathy on the part of the therapist. Vignettes from the author's practice and from the cases presented in this issue are discussed in terms of these five modes of resistance and their treatment. © 2002 John Wiley & Sons, Inc. J Clin Psychol/In Session 58: 157–163, 2002.
An interpersonal communication perspective on resistance in psychotherapy
Authors: Van Denburg Todd F. , Kiesler Donald J.
Published: August 2002

This article discusses resistance in psychotherapy using the contemporary interpersonal communication model of psychotherapy. This perspective defines resistance as moments during sessions when the patient and therapist are interacting with one another in such a way that the patient is kept from becoming aware of any covert experiences or transactional patterns that are conflictual and anxiety provoking. The ways in which resistance may be conceptualized and worked with are discussed and applied to three patient vignettes, with an emphasis on working with resistances as they are manifested in the patient–therapist relationship. Some of the potential reactions of the therapist to resistance are discussed, including some relatively beneficial and some problematic instances. © 2002 John Wiley & Sons, Inc. J Clin Psychol/In Session 58: 195–205, 2002.
Going for the Goal: Improving youths' problem‐solving skills through a school‐based intervention
Authors: O'Hearn Todd C. , Gatz Margaret
Published: August 2002

This study evaluated Going for the Goal (GOAL), a school‐based intervention designed by Danish and colleagues to teach life skills to at‐risk urban adolescents. We extended previous evaluation of GOAL by including an assessment of means‐ends problem‐solving skills. The 10‐week program was administered to 479 middle school students by 46 trained high school student leaders in a predominantly Hispanic community. The program focused on setting positive, reachable goals; anticipating and responding to barriers to goal attainment; using social support; and building on one's strengths. Results demonstrated gains in knowledge of the skills being taught and improvement in problem‐solving skills. Leaders also showed an increase in their knowledge of life skills. The approach maximizes both community resources and ecological validity while giving high school leaders the chance to benefit in their role as helpers. © 2002 Wiley Periodicals, Inc.
A reunion of ideas: Complementary inquiry and collaborative interventions of spirituality, religion, and psychology
Authors: Moore Thom , Kloos Bret , Rasmussen Rachel
Published: August 2002

The intent of this special issue is to extend community psychology's voice in the growing discourse about spirituality, religion, and psychology. While lively debate will continue about particular religious beliefs, the social significance of religious and spiritual experiences cannot be overlooked. These papers will not enter the debate over the existence of a higher power, but rather they will contribute to the development of knowledge and greater understanding of how these phenomena are interwoven into social life. Some authors focus on how religion and spirituality can serve as independent variables that have an impact on phenomena of interest, from the individual to the institution. In other papers, spirituality is treated more as a dependent variable, important as an outcome in its own right. In several of the papers, religion and spirituality are integral parts of an intervention. In sum, when religion and spirituality are viewed as legitimate variables of psychological study, they offer seemingly endless perspectives on human behavior. © 2001 John Wiley & Sons, Inc.
Role of phospholipases A2 in growth‐dependent changes in prostaglandin release from 3T6 fibroblasts
Authors: Sánchez Teresa , Moreno Juan Jose
Published: August 2002

Previously, we reported a growth‐dependent change in prostaglandin production as a consequence of a marked growth‐dependent alteration in arachidonic acid (AA) mobilization from phospholipids. Our present results show that fetal calf serum (FCS) and 4β‐phorbol‐12‐myristate acetate (PMA) caused an enhancement of phospholipase A2 (PLA2) activity in the membrane fraction of non‐confluent cells allowing PLA2 access to its substrate and the release of AA. Western blot analysis has shown that FCS and PMA increased secreted PLA2 (sPLA2) expression in non‐confluent 3T6 fibroblast cultures. Moreover, FCS and PMA induced dithiothreitol‐sensitive and bromoenol lactone‐sensitive PLA2 activities in cytosol and membrane fraction. However, these stimuli did not modify significantly the PLA2 activity in both fractions when 3T6 fibroblasts reached a high cell density. This could be associated with the impairment of AA mobilization in these cell culture conditions. On the other hand, we observed that FCS and PMA induced the same prostaglandin H synthase‐2 induction in non‐confluent and confluent culture conditions. Moreover, the prostaglandin E2 levels reached in cell culture supernatants were independent of the degree of confluence when AA was added exogenously. These results suggest that the changes of intracellular distribution of PLA2 activity of sPLA2 and iPLA2 stimulated by exogenous stimuli may be controlled by cell density conditions which constitute an important mechanism in the regulation of prostaglandin release.© 2001 Wiley‐Liss, Inc.
Acute endotoxemia prolongs the survival of rat lung neutrophils in response to 12‐O‐tetradecanoyl‐phorbol 13‐acetate
Authors: Sunil Vasanthi R. , Connor Agnieszka J. , Lavnikova Natasha , Gardner Carol R. , Laskin Jeffrey D. , Laskin Debra L.
Published: August 2002

Acute endotoxemia is associated with prolonged survival of adherent neutrophils in the lung vasculature. In the present studies, the effects of inflammatory mediators on signaling pathways regulating neutrophil survival were examined. We found that the protein kinase C activator, 12‐O‐tetradecanoyl‐phorbol 13‐acetate (TPA), but not interferon‐γ (IFN‐γ), prolonged the survival of adherent vasculature lung neutrophils from endotoxemic rats, a response that was correlated with reduced apoptosis. Although endotoxin administration to rats induced the expression of the anti‐apoptotic protein Mcl‐1 in lung neutrophils, TPA had no effect on this response. Endotoxin administration also induced expression of total p38 and p44/42 mitogen activated protein kinases (MAPK) in neutrophils, as well as phosphatidyl inositol 3 kinase (PI3K) and its downstream target protein kinase B (PKB). Treatment of the cells with TPA increased p38 MAPK expression in cells from both control and endotoxin treated animals. Cells from endotoxin treated, but not control animals, were found to exhibit constitutive binding activity of nuclear factor kappa B (NF‐κB) which was blocked by TPA. In contrast, constitutive CCAAT/enhancer binding protein (C/EBP) nuclear binding activity evident in neutrophils from control animals was reduced following endotoxin administration. Moreover, this response was independent of TPA. These data suggest that NF‐κB plays a role in TPA‐induced signaling leading to prolonged survival of adherent vascular neutrophils in the lung during acute endotoxemia. J. Cell. Physiol. 190: 382–389, 2002. © 2002 Wiley‐Liss, Inc.
Delaying S‐phase progression rescues cells from heat‐induced S‐phase hypertoxicity
Authors: VanderWaal R.P. , Griffith C.L. , Wright W.D. , Borrelli M.J. , Roti J.L. Roti
Published: August 2002

The mechanism by which a cell protects itself from the lethal effects of heat shock and other stress‐inducing agents is the subject of much research. We have investigated the relationship between heat‐induced damage to DNA replication machinery and the lethal effects of heat shock, in S‐phase cells, which are more sensitive to heat shock than either G1 or G2. We found that maintaining cells in aphidicolin, which prevents the passage of cells through S‐phase, can rescue S‐phase HeLa cells from the lethal effects of heat shock. When S‐phase, HeLa cells were held for 5–6 h in 3 μM aphidicolin the measured clonogenic survival was similar to that for exponentially growing cells. It is known, that heat shock induces denaturation or unfolding of proteins, rendering them less soluble and more likely to co‐isolate with the nuclear matrix. Here, we show that enhanced binding of proteins involved in DNA replication (PCNA, RPA, and cyclin A), with the nuclear matrix, correlates with lethality of S‐phase cells following heat shock under four different experimental conditions. Specifically, the amounts of RPA, PCNA, and cyclin A associated with the nuclear matrix when cells resumed progression through S‐phase correlated with cell killing. Heat‐induced enhanced binding of nuclear proteins involved with other aspects of DNA metabolism, (Mrell, PDI), do not show this correlation. These results support the hypothesis that heat‐induced changes in the binding of proteins associated with DNA replication factories are the potentially lethal lesions, which become fixed to lethal lesions by S‐phase progression but are repairable if S‐phase progression is delayed. © 2001 Wiley‐Liss, Inc.
Effects of nitric oxide and peroxynitrite on endotoxin‐induced leukocyte adhesion to endothelium
Authors: Lelamali Kumtorn , Wang Wei , Gengaro Patricia , Edelstein Charles , Schrier Robert W.
Published: August 2002

Leukocyte accumulation has been shown to be increased in sepsis. Moreover, in inducible nitric oxide synthase (iNOS) knockout mice, a further increase in leukocyte accumulation has been observed during sepsis, suggesting that nitric oxide (NO) may affect leukocyte/endothelial interaction. Accelerated peroxynitrite formation also occurs during sepsis. In the present study, the effect of peroxynitrite or NO on leukocyte adhesion to nitric oxide synthase (NOS)‐inhibited or endotoxin‐treated endothelium was examined. Bovine aortic endothelial cells were treated with either l‐NAME or lipopolysaccharide (LPS) and interferon‐gamma for 4 hr and subsequent leukocyte adhesion was measured. Both l‐NAME and LPS treatment resulted in increased leukocyte adhesion compared with control. Neither a peroxynitrite donor, SIN‐1, nor a direct NO donor, DETA‐NO, had any effect on leukocyte adhesion to untreated endothelium. However, when the l‐NAME or LPS‐treated endothelial cells were treated simultaneously with either SIN‐1 or DETA‐NO, there was a significant reduction in leukocyte adhesion. Moreover, at the concentrations used in the present study, neither peroxynitrite nor NO showed harmful effects on normal cultured endothelial cells. These data demonstrating inhibition of leukocyte adhesion to endotoxin‐treated endothelium suggest that peroxynitrite or NO may exert a beneficial effect during sepsis. © 2001 Wiley‐Liss, Inc.
Influence of some amino acids and vitamin C on the anodic dissolution of tin in sodium chloride solution
Authors: Morad Mohamed Samir S , Hermas Abo Alhagag A
Published: August 2002

The safety of canned foodstuffs should be evaluated not only by recognition of functional foods but also by the presence of toxicants in the human diet. In this study, the anodic dissolution of tin in 3.5% NaCl solution with and without 25–100 mM glycine, serine, methionine, vitamin C and some of their binary mixtures was studied by means of potentiodynamic and impedance techniques. The corroded tin surface was examined by SEM. The results indicated that the passive behaviour of tin is greatly improved by the presence of 50–100 mM glycine and methionine while such improvement is achieved only at 100 mM serine. Both cysteine and vitamin C showed aggressive action. The influence of the glycine/methionine mixture was found to be similar to that of the individual components, whereas the presence of cysteine in the cysteine/methionine mixture annuled the inhibiting action of methionine. The impedance results revealed that the dissolution reaction of tin in NaCl solution with and without glycine, serine and methionine is controlled by the charge transfer process while in the presence of cysteine and vitamin C, the mixed charge transfer and diffusion control is dominant. A rough, bare tin surface was observed in the presence of cysteine and vitamin C while a clean surface covered by corrosion products, in some zones, was observed in NaCl solution containing methionine.

© 2001 Society of Chemical Industry
Enhanced activities of lipase pretreated with organic solvents
Authors: Matsumoto Michiaki , Kida Koji , Kondo Kazuo
Published: August 2002

The catalytic activities of lipases derived from Pseudomonas sp and pretreated with various organic solvents were investigated. The activity of the solvent‐pretreated lipase was greater than that of native lipase in both the esterification reaction in an organic medium and the hydrolysis reaction in an aqueous medium. With esterification calalysed by pretreated lipase, the product, benzyl octanoate, was detected without time‐lag. Conversions at equilibrium state were correlated with the hydrophobicities of the solvents used. In the hydrolysis reaction, most pretreated lipases yielded increased acid production compared with native lipase. A linear correlation was observed between the solvent hydrophobicity and the relative initial reaction rate of the hydrolysis reaction when using pretreated lipases.

© 2001 Society of Chemical Industry
Hereditary hemorrhagic telangiectasia: Demonstration of portosystemic venous shunts using power Doppler sonography and sonographically guided percutaneous transhepatic portovenography
Authors: Lin Chen‐Chun , Wu Jaw‐Ching , Chou Yi‐Hong , Huang Yi‐Hsiang , Chang Full‐Young , Lee Shou‐Dong
Published: August 2002

Hereditary hemorrhagic telangiectasia is a rare autosomal dominant inherited disease associated with vascular abnormalities, which may occur in any organ. Cases of hereditary hemorrhagic telangiectasia accompanied by intrahepatic portosystemic venous shunts, however, have rarely been described. We report a case of hereditary hemorrhagic telangiectasia in which intrahepatic portosystemic shunts were detected using power Doppler sonography and portovenography with percutaneous transhepatic contrast agent injection. On gray‐scale sonography, the common hepatic artery was dilated, and dilated tubular structures mimicking dilated biliary tracts were found. Power Doppler sonography demonstrated the continuity of tortuous vascular channels connecting a branch of the right portal vein to a branch of the right hepatic vein. The dilated vascular channels and tributaries of the right hepatic vein showed a monophasic waveform pattern on spectral analysis. Portovenography showed a tangle of vascular structures connecting with a branch of the right hepatic vein. © 2001 John Wiley & Sons, Inc. J Clin Ultrasound 29:517–522, 2001.
Changes in renal artery resistance after meal‐induced splanchnic vasodilatation in cirrhotic patients
Authors: Perney Pascal , Taourel Patrice , Gallix Benoit , Dauzat Michel , Joomaye Zouberr , Djafari Mounir , Fabro‐Peray Pascale , Beaufort Philippe , Petrissans Bernard , Bruel Jean Michel , Blanc François
Published: August 2002

Purpose

A relationship between vasomotor tone changes in mesenteric and renal vessels in cirrhotic patients has been suspected but remains controversial. The aim of this study was to assess by duplex Doppler sonography the changes in the circulatory resistance of the renal arteries and superior mesenteric artery (SMA) following meal‐induced splanchnic vasodilatation.

Methods

Twenty‐seven cirrhotic patients and 15 healthy volunteers with no hepatic or renal dysfunction were prospectively included in the study. The resistance index (RI) of the SMA and of the right and left renal arteries was measured by duplex Doppler sonography before and 30 minutes after ingestion of a standard 400‐kcal balanced liquid meal. Values in controls and patients and values before and after the meal were compared, and correlations between RIs, Child‐Pugh class (liver function), and creatinine clearance were assessed in cirrhotic patients.

Results

The fasting renal artery RI was greater in cirrhotic patients than in controls (p < 0.0001), but there was no difference in fasting SMA RIs. After the meal, there was a significant decrease in the SMA RI in controls (0.85 ± 0.04 before versus 0.74 ± 0.03 after meal, p = 0.0001) and in cirrhotic patients (0.85 ± 0.04 before versus 0.77 ± 0.04 after, p = 0.0001) and a significant increase in the renal artery RI (0.57 ± 0.06 before versus 0.62 ± 0.05 after in controls, p = 0.001; 0.68 ± 0.07 before versus 0.70 ± 0.07 after in cirrhotic patients, p = 0.001). No correlation was found in cirrhotic patients between the changes in renal artery RI and the postprandial SMA RI decrease, the Child‐Pugh class, or the creatinine clearance.

Conclusions

Meal‐induced SMA vasodilatation (RI decrease) is associated with a marked increase in the renal artery RI, worsening the renal vasoconstriction in cirrhotic patients. © 2001 John Wiley & Sons, Inc. J Clin Ultrasound 29:506–512, 2001.
Prenatal sonographic diagnosis of Majewski syndrome
Authors: Sirichotiyakul Supatra , Tongsong Theera , Wanapirak Chanane , Chanprapaph Pharuhas
Published: August 2002

Majewski syndrome is an autosomal recessive disorder characterized by short ribs, polydactyly, short limbs, and a cleft lip. A 26‐year‐old woman with no family history of genetic diseases presented at 31 weeks' menstrual age with preterm labor and underwent prenatal sonography to screen for fetal anomalies. Sonography revealed a small thorax, markedly short ribs, micromelia, polydactyly, a cleft lip, dolichocephaly, and severe oligohydramnios. The other structures appeared normal. The most likely diagnosis was Majewski syndrome. Vaginal breech delivery was allowed to proceed. The 1,850‐g male newborn died of respiratory failure shortly after birth. The postnatal appearance and radiographs confirmed the prenatal diagnosis. © 2002 Wiley Periodicals, Inc. J Clin Ultrasound 30:303–307, 2002; Published online in Wiley InterScience (www. interscience.wiley.com). DOI: 10.1002/jcu.10066
Confocal microscopy and stereology: Estimating volume, number, surface area and length by virtual test probes applied to three‐dimensional images
Authors: Kubínová Lucie , Janáček Jiří
Published: August 2002

The opportunities of confocal microscopy applied to morphometry of microscopical structures are presented and demonstrated on stereological methods based on evaluation of optical sections within a thick slice and using computer‐generated virtual test probes. Such methods, allowing arbitrary orientation of the thick slice, can be used for estimating volume, number, surface area, and length. The methods using spatial grid of points, disector, fakir, and slicer probes are described and illustrated by different examples using our freeware 3DTOOLS software and their variance and applicability are discussed. It is shown that shifted triple or quadruple spatial grids of lines are very efficient for the surface area and volume estimation by the fakir method. Microsc. Res. Tech. 53:425–435, 2001. © 2001 Wiley‐Liss, Inc.
Apolipoprotein E immunoreactivity in neurons and neurofibrillary degeneration of aged non‐demented and alzheimer's disease patients
Authors: Gómez‐Ramos Pilar , Mufson Elliott J. , Morán M. Asunción
Published: August 2002

Apolipoprotein E (ApoE) genotype is a risk factor for Alzheimer's disease (AD) but its relationship with neurofibrillary degeneration remains obscure. To further analyze this relationship, hippocampal, entorhinal, temporopolar, and insular cortices of 10 non‐demented and 7 Alzheimer disease brains were studied with both light and electron microscopy. Focus was directed on pretangles and neurons starting to accumulate tangles. ApoE immunolabeling in neurons and tangles was independent of ApoE individual genotype. The majority of the neurons in all of the brains were ApoE‐negative, but virtually every brain also contained groups of ApoE‐immunoreactive neurons, with diffuse cytoplasmic labeling. Most of the ApoE‐positive tangles were extracellular, but a few tangles were shown to be intraneuronal when studied ultrastructurally. No ApoE immunoreactivity was found in neuropil threads, as well as in neurites associated with senile plaques. Double protocols with both AT‐8 and anti‐ApoE antibodies, performed to determine whether ApoE‐positive neurons were pretangle neurons, did not detect cytoplasmic AT‐8 in ApoE‐positive neurons. Though careful electron microscopy studies found ApoE reaction product in an occasional ApoE‐positive pretangle‐like neuron and a few intracellular tangles, these findings do not support that ApoE is necessary for the accumulation of hyperphosphorylated tau protein. The more consistent colocalization of anti‐ApoE and AT‐8 in extracellular tangles reveals that ApoE mainly binds to tangles once they are in the extracellular space, in a manner similar to that described for amyloid fibrils. Microsc. Res. Tech. 55:48–58, 2001. © 2001 Wiley‐Liss, Inc.
A conserved family of doublesex‐related genes from fishes
Authors: Huang Xiao , Cheng Hanhua , Guo Yiqing , Liu Li , Gui Jianfang , Zhou Rongjia
Published: August 2002

The sex‐determining gene Mab‐3 of C. elegans and the doublesex gene of Drosophila each contain a common DM domain and share a similar role. Human doublesex‐related gene DMRT1 also encodes a conserved DM‐related DNA‐binding domain. We present here the amplification of a broad range of DM domain sequences from three fish species using degenerate PCR. Our results reveal unexpected complexity of the DM domain gene family in vertebrates. J. Exp. Zool. (Mol. Dev. Evol.) 294:63–67, 2002. © 2002 Wiley‐Liss, Inc.
Development of embryos from in vitro ovulated and fertilized oocytes of the quail (Coturnix coturnix japonica)
Authors: Olszańska Bozdot;enna , Stȩpińska Urszula , Perry Margaret M.
Published: August 2002

The development of quail embryos obtained after in vitro fertilization of oocytes ovulated in vitro was investigated. About 40% of the specimens, after 18–20 hr of incubation, had undergone cleavage to reach stages IV–VI when viewed under a stereo microscope. However, only 36% of these embryos contained normal, DAPI‐stained nuclei when observed under a fluorescent microscope; the other 64% showing a morphologically normal cleavage pattern did not contain nuclei. Control unfertilized oocytes, ovulated in vitro and cultured for the same time, also sometimes attained the morphologically correct stages IV–VI but their “blastomeres” were always devoid of nuclei. Therefore, it is advisable to monitor early avian embryos for the presence of nuclei when assessing development in culture.

The results demonstrate, for the first time, that cytoplasmic segmentation can occur in the absence of nuclear divisions in the germinal disc of the quail and show the existence and significance of ooplasmic maternal information in birds. This phenomenon is also known for sea urchin and frogs. It is indicative of the role of maternal information in early development. The in vitro method presented here links the steps of ovulation and fertilization with the early cleavage stages under in vitro conditions and may be useful in studying mechanisms of fertilization and differentiation in birds as well as in obtaining transgenic birds by DNA injection or application of foreign, DNA‐carrying sperm. J. Exp. Zool. 292:580–586, 2002. © 2002 Wiley‐Liss, Inc.
Gill circulation: regulation of perfusion distribution and metabolism of regulatory molecules
Authors: Olson Kenneth R.
Published: August 2002

The fish gill is the primary regulatory interface between internal and external milieu and a variety of neurocrine, endocrine, paracrine, and autocrine signals coordinate and control gill functions. Many of these messengers also affect gill vascular resistance, and they, in turn, may be inactivated (or activated) by branchial vessels. Few studies have critically addressed how flow is distributed within the gill filament, the physiological consequences thereof, or the impact of gill hormone metabolism on gill and systemic homeostasis. In most fish, the entire cardiac output perfuses the arterioarterial pathway, and this network probably accounts for the majority of passive‐ and stimulus‐induced changes in vascular resistance. The in‐series arrangement of the extensive gill microcirculation with systemic vessels is also indicative of a high capacity for metabolism of plasma‐borne messengers as well as xenobiotics. Adenosine, arginine vasotocin (AVT), and endothelin (ET) are the most potent gill constrictors identified to date, and all decrease lamellar perfusion. Perhaps not surprising, they are also inactivated by gill vessels. Acetylcholine favors perfusion of the alamellar filamental vasculature, although the physiological relevance of acetylcholine‐mediated responses remains unclear. Angiotensin, bradykinin, urotensin, natriuretic peptides, prostaglandins, and nitric oxide are vasoactive to varying degrees, but their effects on intrafilamental blood flow are unknown. If form befits function, then the complex vascular anatomy of the gill suggests a level of regulatory sophistication unparalleled in other vertebrate organs. Resolution of these issues will be technically challenging but unquestionably rewarding. © 2002 Wiley‐Liss, Inc.
Na+, Cl−, Ca2+ and Zn2+ transport by fish gills: retrospective review and prospective synthesis
Authors: Marshall W.S.
Published: August 2002

The secondary active Cl− secretion in seawater (SW) teleost fish gills and elasmobranch rectal gland involves basolateral Na+,K+‐ATPase and NKCC, apical membrane CFTR anion channels, and a paracellular Na+‐selective conductance. In freshwater (FW) teleost gill, the mechanism of NaCl uptake is more controversial and involves apical V‐type H+‐ATPase linked to an apical Na+ channel, apical Cl−–HCO exchange and basolateral Na+,K+‐ATPase. Ca2+ uptake (in FW and SW) is via Ca2+ channels in the apical membrane and Ca2+‐ATPase in the basolateral membrane. Mainly this transport occurs in mitochondria rich (MR) chloride cells, but there is a role for the pavement cells also. Future research will likely expand in two major directions, molded by methodology: first in physiological genomics of all the transporters, including their expression, trafficking, operation, and regulation at the molecular level, and second in biotelemetry to examine multivariable components in behavioral physiological ecology, thus widening the integration of physiology from the molecular to the environmental levels while deepening understanding at all levels. © 2002 Wiley‐Liss, Inc.
Effects of feeding high dosages of vitamin E to laying hens on thyroid hormone concentrations of hatching chicks
Authors: Engelmann Daniela , Flachowsky Gerhard , Halle Ingrid , Sallmann Hans Peter
Published: August 2002

Long‐term experimental feeding of 20,000 ppm α‐tocopheryl acetate to laying hens caused a significant (P < 0.05) decrease in hatching rates as compared to the control group, which was fed a diet containing 19 ppm α‐tocopherol. When the thyroid hormones in the developing chicks were checked on incubation days 16, 19, 21, and 22, the following results were ascertained: During the latter part of incubation, increases in plasma concentrations of thyroxine and triiodothyronine were observed. No significant differences in hormone concentrations (P > 0.05) between the control and the treatment group were observed during incubation days 16, 19, and 22. However, on the day of hatching (day 21 of incubation) significantly lower (P < 0.05) triiodothyronine concentrations in chick embryos of pipped eggs were found in the treatment group. Moreover, thyroxine concentrations in non‐pipped eggs and in hatched chicks were found to be significantly higher as compared to the control group. Given these results, one concludes that extremely high dosages of vitamin E may affect thyroid hormone concentrations of hatching chicks, and therefore, the chicks might be inhibited in pipping the egg shell. Hypothetically, the hepatic enzyme 5′‐monodeiodinase is involved in the mechanism of inhibition. J. Exp. Zool. 290:41–48, 2001. © 2001 Wiley‐Liss, Inc.
Liver function tests during amoebic liver abscess formation in indomethacin‐treated hamsters
Authors: Sánchez‐Ramírez Blanca , Mata‐González Sergio , Valdez Amelia , Ramos‐Martínez Ernesto , Talamás‐Rohana Patricia
Published: August 2002

Establishment of Entamoeba histolytica infection is facilitated through macrophage effector disruption by a prostaglandin E2 (PGE2)‐mediated mechanism. Infection severity may be measured by weight of abscess formed. Indomethacin (Indo) treatment of infected hamsters reduced abscess weight by 30% at 7 days post‐infection presumably by inhibition of PGE2. To explain reductions in abscess development by Indo treatment, we determined liver functionality in Indo‐treated or untreated animals, either healthy or infected. Determinations of serum glutamic oxaloacetic (SGOT) and glutamic pyruvic (SGPT) transaminases, serum alkaline phosphatase (SAP), total serum protein (TSP), and bilirubinemia were done. SGOT, SGPT, and SAP activities showed a significant increase in their values by 600% at seven days post‐infection in infected animals in both conditions; nonstatistical differences were found between animals treated or not. This increase did not correlate with the percentage of damage. Infected nontreated hamsters showed TSP levels 30% below normal group (P < 0.05). Infected Indo‐treated hamsters had no significant differences compared to normal values. Infected nontreated animals showed an increase in bilirubin, particularly in indirect bilirubin, whereas infected Indo‐treated hamsters showed total bilirubin values lower than normals (P < 0.05), with a decrease in direct bilirubin levels. Our results demonstrated that E. histolytica infection in hamsters produces similar abnormalities in liver function as it does in humans, and that the beneficial effect of Indo treatment on amoebic abscess development is not related with an improvement of liver functionality. J. Exp. Zool. 290:201–206, 2001. © 2001 Wiley‐Liss, Inc.
Food chemical cues elicit general and population‐specific effects on lingual and biting behaviors in the lacertid lizard Podarcis lilfordi
Authors: Cooper William E. , Pérez‐Mellado Valentín
Published: August 2002

Actively foraging lizards are capable of identifying prey using only chemical cues sampled by tongue‐flicking, and the relatively few omnivorous and herbivorous lizards tested similarly can identify both animal and plant foods from chemical cues. Whether lizards that eat plants respond to cues specific to preferred plant types and whether there is geographic variability in responses to cues from various plants correlated with the importance of those plants in local diets is unknown. In three populations of an omnivorous lacertid, the Balearic lizard Podarcis lilfordi, we studied chemosensory sampling and feeding responses to chemical cues from plant and animal foods presented on cotton swabs. Each lizard population is endemic to one islet off the coast of Menorca, Balearic Islands, Spain. Lizards in all three populations discriminated chemical cues from plant and animal foods from control substances. Our results extend findings of prey chemical discrimination and plant chemical discrimination in omnivores, increasing confidence that correlated evolution has occurred between plant diet and chemosensory response to palatable plants. There were no consistent differences among populations in tongue‐flicking and biting responses to stimuli from flowers of syntopic and allopatric plant species. The lizards may respond to cues indicative of palatability in a wide range of plant species rather than exhibiting strong responses only to locally available plant species. Nevertheless, tongue‐flicking and biting frequencies varied among plant species, perhaps indicating food preferences. In addition, there were differences among populations in tongue‐flick rates, latency to bite, and licking behavior. Licking was observed in only one lizard population as a response to floral chemicals from only one of the plants species tested, raising the possibility of a population‐specific linkage between identification of a particular plant species and performance of an appropriate feeding response. J. Exp. Zool. 290:207–217, 2001. © 2001 Wiley‐Liss, Inc.
Carbonic anhydrase isozyme distribution and characterization in metabolic fiber types of the dorsal levator muscle of the blue crab, Callinectes sapidus
Authors: Henry Raymond P. , Bilger S. Michelle , Moss Anthony G.
Published: August 2002

Carbonic anhydrase (CA) distribution and characterization were examined in white and light pink fibers of the dorsal levator muscle of the blue crab. White fibers were structurally and metabolically characterized as fast twitch glycolytic, while the light pink fibers were fast oxidative. All subcellular fractions of both fiber types had significant levels of CA activity; cytoplasmic and microsomal activity was significantly higher in light pink vs white fibers. Cytoplasmic CA from both fiber types was highly sensitive to the inhibitors acetazolamide and chlorzolamide, with Ki values of approximately 2 and 0.4 nM, respectively. Further analysis confirmed that cytoplasmic CA from both fiber types was kinetically similar to the high turnover Type II isoform. It appears that the evolution of the CA Type III isoform, found in vertebrate red muscle, did not occur with the differentiation of metabolic fiber types in crustaceans. Membrane‐associated CA, which was also kinetically similar to the Type II isoform, was 20‐fold higher in light pink fibers, suggesting a physiological role in facilitated CO2 efflux from the muscle fiber during periods of prolonged maximal activity. J. Exp. Zool. 290:234–246, 2001. © 2001 Wiley‐Liss, Inc.
Maturation and fertilization in Lottia gigantea oocytes: Intracellular pH, Ca2+, and electrophysiology
Authors: Gould Meredith C. , Stephano José Luis , Ortíz‐Barrón Bernardino de Jesús , Pérez‐Quezada Isela
Published: August 2002

Intracellular pH and Ca2+ were measured with BCECF‐ and Calcium Green‐dextran during maturation and fertilization of oocytes of the limpet Lottia gigantea. Maturation of oocytes from prophase to metaphase I of meiosis was induced in seawater adjusted to pH 9 with NH4OH. Intracellular pH rose during maturation induction, and maturation was also induced by microinjecting pH 8, but not pH 7, HEPES buffer. Intracellular Ca2+ rose during NH4OH‐induced maturation, but maturation was not inhibited when the increase was blocked by microinjection of BAPTA. When the metaphase I oocytes were fertilized , there was an abrupt increase in intracellular Ca2+, and activation (polar body formation) failed to occur in BAPTA‐injected oocytes. Intracellular pH did not rise during fertilization. These observations show that maturation from prophase to metaphase I of meiosis is pH‐dependent and activation of the metaphase I oocytes is Ca2+‐dependent. A Ca2+ action potential was present in both immature and mature oocytes but was more prominent in mature oocytes whose input resistance was higher. Fertilization produced a long‐lasting (17–20 min) Na+‐dependent fertilization potential with superimposed oscillations resembling Ca2+ action potentials. J. Exp. Zool. 290:411–420, 2001. © 2001 Wiley‐Liss, Inc.
Sex determining genes and sexual differentiation in a marsupial
Authors: Pask Andrew , Renfree Marilyn B.
Published: August 2002

The role of genes in the differentiation of the testis and ovary has been extensively studied in the human and the mouse. Despite over a decade of investigations, the precise roles of genes and their interactions in the pathway of sex determination are still unclear. We have chosen to take a comparative look at sex determination and differentiation to gain insights into the evolution and the conserved functions of these genes. To achieve this, we have examined a wide variety of eutherian sex determining genes in a marsupial, the tammar wallaby, to determine which genes have a conserved and fundamental mammalian sex determining role. These investigations have provided many unique insights. Here, we review the recent molecular and endocrine investigations into sexual development in marsupials, and highlight how these studies have shed light on the roles of genes and hormones in mammalian sex determination and differentiation. J. Exp. Zool. 290:586–596, 2001. © 2001 Wiley‐Liss, Inc.
Sry promoters from domesticus (Tirano) and C57BL/6 mice function similarly in embryos and adult animals
Authors: Xian Meiwei , Zhang Jianqing , Lau Yun‐Fai Chris
Published: August 2002

Introduction of the Y chromosome from a Mus musculus domesticus (Tirano) subspecies into the Mus musculus musculus C57BL/6 (B6) inbred strain background results in sex reversal in XY offspring. It has been hypothesized that the domesticus testis‐determining Y (Tdy) locus is misregulated in B6 genome, thereby impairing sex determination in B6.YDom animals. The identification of a gene in the sex‐determining region on the Y chromosome (Sry) as the Tdy has provided a means to experimentally examine this hypothesis. We have generated several lines of B6 transgenic mice harboring a green fluorescent protein gene directed by a Sry promoter from the domesticus (Tirano) Y chromosome. Detailed analysis of the transgene expression was conducted in both fetal and adult tissues of the transgenic mice. The domesticus Sry promoter was capable of directing the expression of the green fluorescent protein gene in a pattern similar, if not identical, to that of the endogenous B6 Sry gene. These observations suggest that the domesticus Sry promoter is not involved in the postulated misregulation of the domesticus (Tirano) Sry gene in the B6 genomic background. These results are discussed with reference to a second hypothesis invoking incompatible protein interaction(s) as a mechanism of aberrant sex determination in B6.YDom animals. J. Exp. Zool. 290:632–641, 2001. © 2001 Wiley‐Liss, Inc.
Strategies for engineering human chromosomes with therapeutic potential
Authors: Saffery Richard , Andy Choo K. H.
Published: August 2002

Human engineered chromosomes (HECs) have several potential advantages over currently used vectors for gene therapy applications. Firstly, there is no upper size limit to DNA that can be cloned in these vectors. Secondly, their extrachromosomal nature ensures that introduced genes are neither disruptive to, nor affected by, the genome of the host cell. Finally, being solely human in origin, HEC vectors should not evoke adverse host immunogenic responses. Recent advances have produced a variety of HECs via several different approaches. This review focuses on the current methodologies for making HEC vectors, the advantages and problems associated with each strategy, and discusses the outlook for HEC vectors as ex vivo therapeutic agents. Copyright © 2001 John Wiley & Sons, Ltd.
Girth of sparse graphs
Authors: Bollobás Béla , Szemerédi Endre
Published: August 2002

For each fixed k ≥ 0, we give an upper bound for the girth of a graph of order n and size n + k. This bound is likely to be essentially best possible as n → ∞. © 2002 Wiley Periodicals, Inc. J Graph Theory 39: 194–200, 2002; DOI 10.1002/jgt.10023