bluuebunny/crossref_metadata_2025 huggingface dataset:
Talsaclidine (WAL 2014 FU), a muscarinic M1receptor agonist for the treatment of Alzheimer's disease
Authors: Wienrich Marion , Ceci Angelo , Ensinger Helmut A. , Gaida Wolfram , Mendla Klaus D. , Osugi Takeshi , Raschig Andreas , Weiser Thomas
Published: October 2002

Since the discovery of muscarinic receptor subtypes, extensive research has been going on attempting to provide treatments for diseases related to malfunctions of cholinergic receptors. In Alzheimer's disease, the prevailing symptomatology is a progressive decline in cognitive functions that can be related to a continuing degeneration of cholinergic neurons. Postsynaptic muscarinic M1receptors, however, do not decline in number during the course of the disease. Talsaclidine is a functionally selective muscarinic M1receptor agonist that has been developed for the symptomatic treatment of Alzheimer's disease. In functional pharmacological assays, talsaclidine was shown to be a full agonist at M1receptors while having only partial agonist activities at M2‐ and M3receptors. In animal studies, this translated into central cholinergic activation measured in rabbits, cats and rats by EEG recordings, and in an improvement of performance in learning and memory tests in aged animals or in animals with experimentally‐induced cholinergic hypofunction. In these studies, cholinergic side effects became apparent in doses higher than those inducing cognitive improvement. In addition to effects increasing cognitive functions, stimulation of M1receptors that belong to the class of G protein‐coupled, seven transmembrane receptors may have a positive impact on the progression of the disease. Talsaclidine was shown to increase the secretion of the neurotrophic ectodomain of the amyloid precursor protein while simultaneously decreasing the formation of the amyloid β (Aβ) protein that constitutes the amyloid plaques, one of the neuropathological hallmarks of the disease. In Alzheimer patients, talsaclidine decreased the concentration of Aβ in the cerebrospinal fluid. Despite these encouraging data, talsaclidine failed to improve cognitive symptoms when Alzheimer patients were treated for 3 months. This confirms reports from other muscarinic agonists, none of which has convincingly demonstrated clinical efficacy, and points to the fact that either the pharmacological profile of current M1receptor agonists still needs improvement or that muscarinic receptor stimulation alone does not suffice to improve the condition of Alzheimer patients. Drug Dev. Res. 56:321–334, 2002. © 2002 Wiley‐Liss, Inc.
Use of vitamin C and E in the treatment of Alzheimer's disease
Authors: Arlt Sönke , Müller‐Thomsen Tomas , Beisiegel Ulrike
Published: October 2002

Since it has become evident that increased oxidative stress is involved in the pathogenesis of Alzheimer's disease, the therapeutical use of antioxidants has been discussed. To date there are several studies showing that lipid peroxidation in particular is increased not only in brain tissue but also in plasma and cerebrospinal fluid of Alzheimer's disease patients. Accordingly, decreased levels of antioxidants, which are capable of inhibiting lipid peroxidation, have been found in a majority of studies performed on this subject. This article focusses on the two major lipid peroxidation inhibiting vitamins, vitamin C (ascorbate) and vitamin E (α‐tocopherol), in the therapy of Alzheimer's disease. The antioxidant action of these vitamins has been investigated in detail in artherosclerosis research, and the combination of both antioxidants has been shown to efficiently inhibit lipoprotein oxidation in plasma and cerebrospinal fluid following oral supplementation. The only double‐blind placebo‐controlled study that has been performed on the effect of vitamin E alone demonstrated a beneficial effect on the course of Alzheimer's disease. At present there is evidence supporting the combined use of vitamin C and E therapy for Alzheimer's disease, but future clinical studies investigating the effect of these antioxidants must be carried out. Drug Dev. Res. 56:452–457, 2002. © 2002 Wiley‐Liss, Inc.
Statistical verification of the Chesapeake Bay benthic index of biotic integrity
Authors: Alden Raymond W. , Dauer D. M. , Ranasinghe J. A. , Scott L. C. , Llansó R. J.
Published: October 2002

The benthic index of biotic integrity (B‐IBI) developed for the Chesapeake Bay was statistically verified using simulations and a suite of multivariate statistical techniques. The B‐IBI uses a simple scoring system for benthic community metrics to assess benthic community health and to infer environmental quality of benthic habitats in the Bay. Overall, the B‐IBI was verified as being sensitive, stable, robust and statistically sound. Classification effectiveness of the B‐IBI increased with salinity, from marginal performance for tidal freshwater ecosystems to excellent results for polyhaline areas. The greater classification uncertainty in low salinity habitats may be due to difficulties in reliably identifying naturally unstressed areas or may be due to regional ecotones created by stress gradients. Pollution‐indicative species abundance, pollution‐sensitive species abundance, and diversity(Shannon's index) were the most important metrics in discriminating between degraded and non‐degraded conditions in the majority of the habitats. Single metrics often performed as well as the multi‐metric B‐IBI in correctly classifying the relative quality of sites. However, the redundancy in the multi‐metric B‐IBI provided a stable ‘weight of evidence’ system which increased confidence in general conclusions. Confidence limits developed for the B‐IBI scores were used to distinguish among habitats that were degraded, non‐degraded, of intermediate quality, or of indeterminate condition. Copyright © 2002 John Wiley & Sons, Ltd.
A cascadic conjugate gradient algorithm for mass conservative, semi‐implicit discretization of the shallow water equations on locally refined structured grids
Authors: Bonaventura Luca , Rosatti Giorgio
Published: October 2002

A semi‐implicit, mass conservative discretization scheme is applied to the two‐dimensional shallow water equations on a hierarchy of structured, locally refined Cartesian grids. Different resolution grids are fully interacting and the discrete Helmholtz equation obtained from the semi‐implicit discretization is solved by the cascadic conjugate gradient method. A flux correction is applied at the interface between the coarser and finer discretization grids, so as to ensure discrete mass conservation, along with symmetry and diagonal dominance of the resulting matrix. Two‐dimensional idealized simulations are presented, showing the accuracy and the efficiency of the resulting method. Copyright © 2002 John Wiley & Sons, Ltd.
Structured multigrid agglomeration on a data structure for unstructured meshes
Authors: Hannemann V.
Published: October 2002

A key issue for using multigrid as an acceleration technique on unstructured meshes is the generation of the coarse grid levels. Some new ideas are presented to improve the agglomeration of coarse cells in hybrid grids with respect to the structure of hexahedral or prismatic layers. The algorithms are embedded in the standard advancing front method which works locally on the edge based data structure representing the dual grid. The goal is to preserve the topological structure of the hexahedral and prismatic parts as far as possible. As a result of the structure preservation a desired coarsening ratio (number of fine grid volumes per coarse grid volume), responsible for the memory requirements and the computing time spent in looping over all volumes, can be achieved with a minimal number of edges, which is linked to the major working time spent in loops over all edges. Copyright © 2002 John Wiley & Sons, Ltd.
Adjoint shape optimization for steady free‐surface flows
Authors: van Brummelen E. H. , Segal A.
Published: October 2002

Numerical solution of flows that are partially bounded by a freely moving boundary is of great importance in practical applications such as ship hydrodynamics. Free boundary problems can be reformulated into optimal shape design problems, which can in principle be solved efficiently by the adjoint method. This work examines the suitability of the adjoint shape optimization method for solving steady free‐surface flows. It is shown that preconditioning is imperative to avoid mesh‐width dependence of the convergence behaviour. Numerical results are presented for 2D flow over an obstacle in a channel. Copyright © 2002 John Wiley & Sons, Ltd.
Somatic mitochondrial DNA mutations in human chromophobe renal cell carcinomas
Authors: Nagy Anetta , Wilhelm Monica , Sükösd Farkas , Ljungberg Börje , Kovacs Gyula
Published: October 2002

We sequenced the entire mitochondrial genome in 8 chromophobe renal cell carcinomas (RCCs) and corresponding normal kidneys. Our study disclosed 68 known and 45 new sequence variations occurring 132 and 45 times, respectively. We found 6 somatic nucleotide changes in 5 out of the 8 chromophobe RCCs. One A → T substitution occurred in the D‐loop region and an insertion of a 9‐bp sequence in the noncoding region of the MTNC7. One G → A substitution and one C → T substitution were seen in the MTRNR1 and MTRNR2 genes, respectively. One C deletion in MTND5 and one T insertion in the MTND3 gene resulted in frameshift mutations in two tumors. All somatic alterations, with the exception of the 9‐bp insertion, were heteroplasmic changes. Although somatic mtDNA mutations are found in chromophobe RCCs, their role in the maintenance of tumor cell phenotype or in tumorigenesis remains to be elucidated. © 2002 Wiley‐Liss, Inc.
The rock‐hewn churches of Tigrai, Northern Ethiopia: A geological perspective
Authors: Asrat Asfawossen
Published: October 2002

Ethiopia is endowed with many rock‐hewn churches, with the largest group in central and eastern Tigrai. These churches can be divided into four zones: the Atsbi, Hawzen‐Ger'alta, Sinkata‐Adigrat, and Tembien, with more than 100 rock‐hewn churches of different ages, sizes, and histories. However, they have one thing in common: All are carved into sandstone. The Enticho, Adigrat, and Ambaradam sandstones (Permo‐Carboniferous, Triassic—Middle Jurassic, and Early Cretaceous, respectively) are extensively exposed in these areas and are characterized by thick and massive beds of coarse‐ to fine‐grained and well‐sorted successions. These sandstones are easily carved, yet compact enough to withstand pressure. Although limestone, basalt, and crystalline rocks are exposed in the same area, few rock‐hewn churches have been carved into them. The rock type is, therefore, the most important factor in the location of these rock‐hewn churches. © 2002 Wiley Periodicals, Inc.
Differential expression of protein tyrosine kinase genes during microglial activation
Authors: Krady J. Kyle , Basu Anirban , Levison Steven W. , Milner Robert J.
Published: October 2002

Protein tyrosine kinase (PTK) activity is abundant in microglia, but the PTKs that participate in their activation have not been identified. For these studies, we used three paradigms to characterize PTK expression during microglial activation: resting and activated microglia were bulk fractionated from the adult brain, cultured newborn microglia were treated with lipopolysaccharide (LPS) to model the transition from activated toward phagocytic microglia, and PTK expression was examined in activated microglia in situ after facial nerve axotomy. Two PCR‐based strategies were used to show that 21 different PTK genes are expressed by rat brain microglia: 5 receptor PTKs, 10 nonreceptor PTKs, and 6 members of the src family. Seven of the 21 PTKs were examined in greater detail. Five PTK mRNAs (fgr, hck, fak, jak‐2, and flk‐1) increased expression across all three models of activation. We conclude that they represent key components in the cascades that participate in microglial activation. In contrast, expression of fes and fms correlated with stimuli that affect microglial proliferation. Four of the PTKs (hck, fgr, fes, and fms) are believed to be myeloid cell specific and were not expressed by cultured astrocytes. HCK and FAK protein were also not expressed in lysates of immature astrocytes and oligodendrocytes. Because of their putative specificity, these kinases represent potential targets for inhibitors of microglial activation. Because reactive microglia can exacerbate the severity of neurological diseases, the identification of specific kinases that participate in microglial activation represents an important advance toward the development of new therapeutics. GLIA 40:11–24, 2002. © 2002 Wiley‐Liss, Inc.
The relative number of macrophages/microglia expressing macrophage colony‐stimulating factor and its receptor decreases in multiple sclerosis lesions
Authors: Werner Katrin , Bitsch Andreas , Bunkowski Stephanie , Hemmerlein Bernhard , Brück Wolfgang
Published: October 2002

The activation of macrophages/microglia in multiple sclerosis (MS) lesions plays a central role in the effector phase of myelin breakdown. The precise patterns of macrophage/microglia activation during demyelination have not yet been defined. The growth and activating factor macrophage‐colony stimulating factor (M‐CSF) and its specific receptor (M‐CSFR) may be involved in this process. The present study investigated the expression of M‐CSF and M‐CSFR mRNA by in situ hybridization in 60 lesions from 32 MS patients. In the control and periplaque white matter, microglia was almost completely M‐CSFR positive. Irrespective of the demyelinating activity, an increased number of cells expressed M‐CSF or M‐CSFR mRNA within the lesions. However, despite the tremendous increase in macrophages/microglia within the lesions, the relative number of these cells expressing M‐CSF or M‐CSFR decreased. There was no correlation of M‐CSF or M‐CSFR expression with active myelin breakdown. The correlation between the clinical course and the expression of M‐CSF or M‐CSFR mRNA revealed significant differences with the lowest expression in primary progressive MS. These results suggest a downregulation of M‐CSF and M‐CSFR inside the MS plaque probably due to the high amount of macrophage‐derived cytokines or mediators. Nevertheless, the differences in the relative number of cells expressing the M‐CSF/M‐CSFR pathway implicate that this pathway may be an important contributory factor in different forms of MS pathology. GLIA 40:121–129, 2002. © 2002 Wiley‐Liss, Inc.
Cognitive function and caregiver burden: predictive factors for eating behaviour disorders in Alzheimer's disease
Authors: Rivière Stéphanie , Gillette‐Guyonnet Sophie , Andrieu Sandrine , Nourhashemi Fati , Lauque Sylvie , Cantet Christelle , Salva Antoni , Frisoni Giovanni , Vellas Bruno
Published: October 2002

Background

Patients with Alzheimer's disease (AD) commonly develop aversive feeding behaviours. These behaviours lead to weight loss and frequently to physicians using tube feeding. Little is known about factors correlated with feeding difficulties during AD.

Objective

To investigate predictors of aversive feeding behaviours (AFBs) which occured during a one‐year interval among AD patients living at home with a caregiver.

Method

One‐year study initiated in January 1998 with 224 AD patients and their caregivers. Main outcome measures: all patients underwent a nutritional, psychologic and functional evaluation at baseline and one year later. Feeding difficulties were assessed using the Feeding Dependency Scale and the Aversive Feeding Behaviour Inventory. The Burden Interview was also done to assess the material and emotional burden.

Results

Initial feeding difficulties were significantly associated with the age of the caregiver, the severity of the disease and the initial patient's autonomy and psychological functioning (mood and behaviour disorders). AD patients, who lived with a more affected caregiver at baseline, developped feeding difficulties and AFBs during the follow‐up. Logistic regression analysis showed a positive association between AFBs worsening and the initial caregiver's burden after controlling for counfounding factors. On the other hand, memory impairment was inversely associated with AFBs.

Conclusions

Both cognitive impairment and family stress can help in predicting which AD patients living at home will develop AFBs. Nutritional information and support to families are probably the best strategies to prevent AFBs during AD and to improve consequently the patient's and caregiver's quality of life. Copyright © 2002 John Wiley & Sons, Ltd.
Carcinoma in situ in a lateral cervical cyst
Authors: Zimmermann Corinna E. , von Domarus Helmut , Moubayed Pierre
Published: October 2002

Background

The development of an invasive squamous cell carcinoma within a lateral cervical cyst as a result of malignant transformation of the epithelium is considered a rare circumstance. The existence of this entity is a source of controversy in light of the differential diagnosis, which includes a cervical metastasis from an unknown primary carcinoma (CUP‐syndrome). Apart from site, histologic findings, and follow‐up, the principal diagnostic criterion for lateral cervical cyst carcinoma is the histomorphologic demonstration of transition of the benign epithelium into invasive squamous cell carcinoma. Although numerous case reports of this entity exist, carcinoma in situ in a lateral cervical cyst has been reported in only five cases thus far.

Method

In this context, we present the case of a 44‐year‐old patient with a 7‐month history of cervical swelling.

Results

After diagnostic extirpation of the tumor, histologic findings were consistent with a lateral branchial cyst with high‐grade dysplasia and carcinoma in situ of the squamous epithelial lining.

Conclusion

This case of a premalignant lesion within a lateral branchial cyst (ie, carcinoma in situ) supplies the “missing link” and adds credibility to the concept of primary branchiogenic carcinoma. © 2002 Wiley Periodicals, Inc.
Gene therapy for intraperitoneally disseminated pancreatic cancers by Escherichia coli uracil phosphoribosiltransferase (UPRT) gene mediated by restricted replication‐competent adenoviral vectors
Authors: Oonuma Masaru , Sunamura Makoto , Motoi Fuyuhiko , Fukuyama Shouji , Shimamura Hiromune , Yamauchi Jun‐ichiro , Shibuya Kazuhiko , Egawa Shin‐ichi , Hamada Hirofumi , Takeda Kazunori , Matsuno Seiki
Published: October 2002

Although patients with unresectable pancreatic tumors have been treated with 5‐fluorouracil (5FU)‐based combination chemotherapy, the drug resistance of cancer cells presents a crucial therapeutic problem. It was reported that UPRT overcomes 5FU resistance. UPRT catalyzes the synthesis of 5‐fluorouridine monophosphate (FUMP) from Uracil and phosphoribosylpyrophosphate (PRPP). The antitumor effect of 5FU is enhanced by augmenting 5‐fluorodeoxyuridine monophosphate (FdUMP) converted from FUMP, which inhibits thymidylate synthetase (TS). We first demonstrated that injecting an E1‐deficient adenoviral vector (Adv) expressing UPRT (AxCAUPRT) followed by 5‐FU treatment resulted in a volume reduction of xenotransplanted human tumors. In examining the therapeutic effect of AxCAUPRT/5‐FU against peritoneal dissemination, we found that non‐selective gene transduction of AxCAUPRT caused severe adverse effects arising from the increase of F‐dUMP in normal intestine. Because the therapeutic gene delivered by a restricted replication‐competent Adv lacking 55 kDa E1B protein (AxE1AdB) is speculated to be expressed selectively in tumors, mice with established tumors were injected with AxE1AdB and E1‐deleted Adv expressing the lacZ reporter gene (AxCAlacZ). The expression of the reporter gene (lacZ) was selectively enhanced in disseminated tumors. The therapeutic advantage of restricted replication competent Adv that expresses UPRT (AxE1AdB‐UPRT) was evaluated in an intraperitoneal disseminated tumor model. To study the anti‐tumor effect of AxE1AdB‐UPRT/5FU, mice with disseminated AsPC‐1 tumors were administered the Adv, followed by the 5FU treatment. It was shown that the treatment with AxE1AdB‐UPRT/5FU caused a dramatic reduction of the disseminated tumor burden without toxicity in normal tissues. Our results showed that the AxE1AdB‐UPRT/5FU system is a promising tool for intraperitoneal disseminated pancreatic cancer. © 2002 Wiley‐Liss, Inc.
Human dendritic cells induce tumor‐specific apoptosis by soluble factors
Authors: Joo Hong‐Gu , Fleming Timothy P. , Tanaka Yoshiyuki , Dunn Toby J. , Linehan David C. , Goedegebuure Peter S. , Eberlein Timothy J.
Published: October 2002

Dendritic cells (DCs) are the most potent antigen producing cells (APCs) for initiation of immune responses including anti‐tumor immune responses. In previous reports, it has been shown that DCs efficiently take up and process apoptotic or necrotic bodies of tumor cells. It has also been shown that DCs pulsed with tumor cell apoptotic bodies, lysates or peptides generate potent anti‐tumor immune responses. Direct interactions between DCs and viable tumor cells, however, have not been clearly elucidated. We report that monocyte‐derived, CD1a+ immature DCs (iDCs) significantly inhibit the growth of breast tumor cells in coculture and transwell experiments in the presence of soluble CD40 ligand (sCD40L), LPS or both. The growth inhibition effects correlated with cell cycle arrest and apoptosis of breast tumor cells. The effects were associated with morphological changes of tumor cells from a round shape to a flat, spindle shape. In contrast, no inhibition of proliferation or morphological changes was observed on normal PBMC, K562 or breast fibroblasts. Interestingly, iDCs undergoing maturation induced by sCD40L+LPS induced a much stronger growth inhibitory effect than iDCs alone or mature DCs treated with sCD40L+LPS. Fractionation of supernatants showed the anti‐tumor effects were mediated by a TNF‐α‐dependent and ‐independent mechanism. Soluble FasL and TRAIL were not involved. Our findings suggest that maturing DCs have the intrinsic ability to induce cell‐cycle arrest and apoptosis of breast tumor cells through soluble factors, but not normal cells, in addition to their Ag presentation function. © 2002 Wiley‐Liss, Inc.
Prolactin secretion and intracellular Ca2+ change in rat lactotroph subpopulations stimulated by thyrotropin‐releasing hormone
Authors: Huang Chih‐Yang , Kuo Wei‐Wen , Tsai Tsung‐Po , Wu Der‐Jinn , Hsieh Yih‐Shou , Wang Paulus S. , Cheng Chih‐Kai , Liu Jer‐Yuh
Published: October 2002

Thyrotropin‐releasing hormone (TRH) may stimulate lactotrophs to increase intracellular Ca2+ and to secrete prolactin (PRL). In this study, PRL contents in lactotrophs were determined by the sequential cell immunoblot assay (SCIBA) and their changes in intracellular Ca2+ was analyzed by confocal microscopy. Significant correlations were found in the corresponding parameters between TRH treatments with a recovery interval of 2 h. Measuring the PRL contents after the first TRH treatment and then determining the intracellular Ca2+ changes after the second TRH treatment revealed four lactotroph subpopulations. Type I cells (51%) showed significant responses of both PRL secretion and intracellular Ca2+ concentration. Type II cells (22%) increased in PRL secretion, but without changes in intracellular Ca2+. Type III cells (17%) have increased in intracellular Ca2+, but without changes in PRL secretion. Type IV cells (10%) did not show changes in PRL secretion and intracellular Ca2+. J. Cell. Biochem. 87: 126–132, 2002. © 2002 Wiley‐Liss, Inc.
Biochemical characterization of heparan sulfate derived from murine hemopoietic stromal cell lines: A bone marrow‐derived cell line S17 and a fetal liver‐derived cell line AFT024
Authors: Arcanjo Katia , Belo Gisele , Folco Cristiane , Werneck Claudio C. , Borojevic Radovan , Silva Luiz‐Claudio F.
Published: October 2002

Heparan sulfate (HS) present on the surface of hemopoietic stromal cells has important roles in the control of adhesion and growth of hemopoietic stem and progenitor cells. Recent studies have characterized several different heparan sulfate proteoglycans (HSPGs) from both human and murine bone marrow stromal cells. In the present study, we have compared the molecular structure of HS, metabolically labeled with [35S]‐sulfate produced by two distinct preparations of murine hemopoietic stromal cell lines. These comprised a bone marrow‐derived cell line S17 and a fetal liver‐derived cell line AFT024. [35S]‐HS was examined in the cell layers and in the culture medium. We identified and measured the relative proportions of the various glycosaminoglycans (GAGs) in the two stromal cell lines. Chondroitin sulfate (CS) was preponderantly secreted by the stromal cell lines, while HS was relatively more abundant in the cell‐associated fractions. The two types of stromal cells differ in their HS composition, mainly due to different patterns of N‐ and O‐sulfation. The two stromal cell lines expressed mRNA for different HSPGs. Data from reverse transcription PCR revealed that the two stromal cell lines expressed mRNA for glypican and syndecan4. Only AFT024 cell line expressed mRNA for betaglycan. There was no evidence for expression of mRNA for both syndecan1 and syndecan2. [35S]‐sulfated macromolecules could be released from the cell surface of both stromal cell lines by phosphatidylinositol phospholipase C (PI‐PLC), which is consistent with the expression of glypican detected by PCR experiments. J. Cell. Biochem. 87: 160–172, 2002. © 2002 Wiley‐Liss, Inc.
Follow‐up of anti‐Aspergillus IgG and IgA antibodies in bone marrow transplanted patients with invasive aspergillosis
Authors: Centeno‐Lima S. , de Lacerda J.M. , do Carmo J.A. , Abecasis M. , Casimiro C. , Exposto F.
Published: October 2002

A total of 89 patients at risk for, or with invasive aspergillosis (IA) were recruited from bone marrow transplantation (BMT) units in two Lisbon hospitals, and followed for 2½ years to monitor their immune response. Of these patients, six developed probable IA, from which five died. The presence of serum IgG or IgA antibodies against seven Aspergillus recombinant antigens was assessed in patients with IA, using an enzyme‐linked immunosorbent assay (ELISA). In parallel, the serum levels of galactomannan (GM) were also monitored, using the Platelia Aspergillus kit (Sanofi Pasteur, Marnes‐la‐Coquette, France). Superoxide dismutase (Sod) and 94 kDa were the most immunogenic antigens for IgA, while the IgG pattern of recognition changed from patient to patient. From our results we conclude that although follow‐up of antibodies against these antigens should not be used as a diagnostic method, patients with IA do produce an immune response that may influence disease outcome. J. Clin. Lab. Anal. 16:156–162, 2002. © 2002 Wiley‐Liss, Inc.
Ethanol production from raw starch by a recombinant yeast having saccharification and fermentation activities
Authors: Nakamura Yoshitoshi , Sawada Tatsuro , Komatsu Akihiro
Published: October 2002

In order to develop a method for converting raw starch into ethanol efficiently, direct fermentation of ozonized raw starch using a recombinant yeast was investigated. Ozonolysis was carried out as a pretreatment to convert raw starch into ethanol rapidly and efficiently, and then the effect of the ozone degradation conditions on the degree of polymerization and the amount of amylose in a raw starch was determined. Since the degree of polymerization was low and the amount of amylose was high, raw starch treated with an ozone concentration of 40 gm−3 and an ozonation time of 30 min was the material chosen for alcohol fermentation. Though the recombinant yeast could not convert the untreated raw starch, it converted the soluble starch and the ozonized raw starch at a comparatively high yield into ethanol. About 56% of the ozonized raw starch decomposed, and the ethanol concentration obtained from the ozonized raw starch was markedly greater than that obtained from untreated raw starch. The dynamic behavior of cell growth, substrate degradation, and ethanol production was examined in a continuous culture under various dilution rates, and the optimal dilution rate, ie 0.15 h−1, was determined for maximizing the ethanol productivity (amount of ethanol produced per unit time).

© 2002 Society of Chemical Industry
Online monitoring of a bioprocess based on a multi‐analyser system and multivariate statistical process modelling
Authors: Cimander Christian , Mandenius Carl‐Fredrik
Published: October 2002

Multivariate statistical process control (MSPC) was for the first time applied to analyse data from a bioprocess on‐line multi‐analyser system consisting of an electronic nose (EN), a near‐infrared spectroscope (NIRS), a mass spectrometer (MS) and standard bioreactor probes. One hundred and fifty sensor signals from the electronic nose, 1050 wavelength signals from the NIRS, carbon dioxide evolution rate calculated from mass spectrometer signals and standard bioreactor data (eg amount of substrate fed) were interrogated for their ability to model a bioprocess using MSPC. The models obtained were validated on a recombinant Escherichia coli fed‐batch process for tryptophan production. Limiting trajectories were defined in the MSPC models for warning, action, and process experience with respect to biomass and tryptophan concentrations. The results showed the capacity and robustness of MSPC models for monitoring with multi‐analysers and allowed a comparison of the different analysers' suitability for this kind of data processing. Furthermore, the results demonstrate that MSPC models provide a functional and versatile framework for coping with large information flows and are also suited to a variety of other bioprocessing monitoring and control tasks.

© 2002 Society of Chemical Industry
An integrated downstream processing strategy for the recovery and partial purification of penicillin acylase from crude media
Authors: Fonseca L P , Cabral J M S
Published: October 2002

An integrated process strategy for the recovery of penicillin acylase was developed, based on precipitation of non‐enzymatic proteins directly from Escherichia coli homogenates or crude extracts using Rolquat (quaternary ammonium salt) and adsorption of the enzyme on Amp‐Seph (3.8 µmole ampicillin cm−3) under pseudo‐affinity conditions. The effect of pH, concentrations of ammonium sulfate and Rolquat, and also concentrations of protein and cell debris on the precipitation of non‐enzymatic proteins from homogenates and crude extracts of penicillin acylase were analysed. The method of addition of Rolquat to homogenates and crude extracts significantly influenced the size of the precipitated particles. Improved results on the specific activity of penicillin acylase were obtained for 22% and 1% (w/v) of ammonium sulfate and Rolquat, respectively, added sequentially to enzyme solutions and at room temperature. Under these experimental conditions, the specific activity of penicillin acylase in homogenates and crude extracts was enhanced 2.5–3.0‐fold. Finally, the integrated process strategy was implemented first by precipitation of non‐enzymatic proteins and recovery of penicillin acylase directly from the enzyme solution treated with Rolquat using an adsorption/filtration system with an overall yield of 86%. This system allows simultaneously the filtration of cell debris and fine precipitated particles, in situ recovery of penicillin acylase by its adsorption on Amp‐Seph, and selective desorption of the enzyme with a specific activity of 11 IU (mg prot)−1 and a desorption yield of 95%.

© 2002 Society of Chemical Industry
Melanization stimulating factor (MSF) and melanization inhibiting factor (MIF) in the integument of fish
Authors: Zuasti Adelina
Published: October 2002

The present studies were directed to demonstrate that adult fish skin contains putative factors that affect chromatophore and/or chromatoblast function. This hypothesis is based upon the possibility that hypo and/or hyperpigmented areas of the skin are so pigmented because of the localized expression of intrinsic factors that are either stimulatory or inhibitory to the differentiation of specific pigment cell types. In all the morphological and biochemical experiments carried out, we used culture media conditioned by dorsal (DCM) or ventral (VCM) skin from different species of fish. Both DCM and VCM were capable of stimulating differentiation of melanophores in neural crest explants. While the stimulation of melanization is an activity present in both dorsal and ventral skin, an inhibitory activity is also present in ventral skin at such a concentration that it overrides the stimulatory activity afforded by DCM. With biochemical assays, we demonstrated that the three important sequential enzymatic steps in melanogenesis are all stimulated by the conditioned media in a dose‐dependent manner and this results in an increase in the amount of melanin present in cultured cells. The results of our investigations provide strong evidence that there are intrinsic pigment cell regulatory factors in the integument of fish, the inhibitory activity being stronger in the ventrum, and that those factors strongly influence, perhaps even determine, the pigment patterns of fish. Microsc. Res. Tech. 58:488–495, 2002. © 2002 Wiley‐Liss, Inc.
Calponin, caldesmon, and chromatophores: The smooth muscle connection
Authors: Meyer‐Rochow Victor Benno , Royuela Mar
Published: October 2002

Observations on pigment translocations in fish chromatophores and speculations on the chemo‐mechanical transduction processes responsible for the recorded chromatosome motilities are briefly reviewed. The presence of the two smooth muscle proteins caldesmon and calponin is confirmed by immunocytochemistry for melanophores and iridophores of the Antarctic fishes Pagothenia borchgrevinki and Trematomus bernacchii. Troponin, a typical vertebrate skeletal muscle protein is absent from the chromatophores of the two fish species. It is suggested that calponin's role, in the presence of Ca2+ and calmodulin, is that of a modulator and that caldesmon, a molecule that competes with calponin for actin binding sites, is in a position in which it can switch on and off Ca2+‐dependent contractility and relaxation. Freshly caught Antarctic fish are receiving conflicting signals, when hauled from the dark under‐ice to the bright above‐ice environment (nor‐adrenaline secretion promoting aggregation, but exposure to bright light bringing on pigment dispersion); it is in such situations that the two proteins in question could play important roles. The precise nature of their involvement still needs to be worked out, but the fact that they do exist in the chromatophores at all, appears to have an ontogenetic background. Microsc. Res. Tech. 58:504–513, 2002. © 2002 Wiley‐Liss, Inc.
Highlight: BRCA1 and BRCA2 proteins in breast cancer
Authors: Daniel Dianne C.
Published: October 2002

Heterozygous carriers of loss‐of‐function germline mutations in the BRCA1 or BRCA2 breast cancer susceptibility genes have a predisposition to breast and ovarian cancer. Multiple functions have been ascribed to the products of these genes, linking them to pathways that inhibit progression to neoplasia. Various investigators have assigned roles for these tumor suppressor gene products in the cell functions of genome repair, transcription, and growth control. There is emerging evidence that BRCA1 may participate in ubiquitin E3 ligase activity. BRCA1 and BRCA2 have each been implicated in chromatin remodeling dynamics via protein partnering. Ubiquitin ligase and chromatin remodeling activities need not be mutually exclusive and both may function in DNA repair, transcriptional regulation, or cell cycle control. Here we highlight certain recent findings and currently unanswered questions regarding BRCA1 and BRCA2 in breast cancer. Microsc. Res. Tech. 59:68–83, 2002. © 2002 Wiley‐Liss, Inc.
Evolutionary divergence of vertebrate Hoxb2 expression patterns and transcriptional regulatory loci
Authors: Scemama Jean‐luc , Hunter Michael , Mccallum Jeff , Prince Victoria , Stellwag Edmund
Published: October 2002

Hox gene expression is regulated by a complex array of cis‐acting elements that control spatial and temporal gene expression in developing embryos. Here, we report the isolation of the striped bass Hoxb2a gene, comparison of its expression to the orthologous gene from zebrafish, and comparative genomic analysis of the upstream regulatory region to that of other vertebrates. Comparison of the Hoxb2a gene expression patterns from striped bass to zebrafish revealed similar expression patterns within rhombomeres 3, 4, and 5 of the hindbrain but a notable absence of expression in neural crest tissues of striped bass while neural crest expression is observed in zebrafish and common to other vertebrates. Comparative genomic analysis of the striped bass Hoxb2a–b3a intergenic region to those from zebrafish, pufferfish, human, and mouse demonstrated the presence of common Meis, Hox/Pbx, Krox‐20, and Box 1 elements, which are necessary for rhombomere 3, 4, and 5 expression. Despite their common occurrence, the location and orientation of these transcription elements differed among the five species analyzed, such that Krox‐20 and Box 1 elements are located 3′ to the Meis, Hox/Pbx elements in striped bass, pufferfish, and human while they are located 5′ of this r4 enhancer in zebrafish and mouse. Our results suggest that the plasticity exhibited in the organization of key regulatory elements responsible for rhombomere‐specific Hoxb2a expression may reflect the effects of stabilizing selection in the evolution cis‐acting elements. J. Exp. Zool. (Mol. Dev. Evol.) 294:285–299, 2002. © 2002 Wiley‐Liss, Inc.
Signed degree sequences and multigraphs
Authors: Michael T. S.
Published: October 2002

We give necessary and sufficient conditions for the existence of a signed r‐multigraph with a prescribed signed degree sequence. © 2002 Wiley Periodicals, Inc. J Graph Theory 41: 101–105, 2002
On a list coloring conjecture of Reed
Authors: Bohman Tom , Holzman Ron
Published: October 2002

We construct graphs with lists of available colors for each vertex, such that the size of every list exceeds the maximum vertex‐color degree, but there exists no proper coloring from the lists. This disproves a conjecture of Reed. © 2002 Wiley Periodicals, Inc. J Graph Theory 41: 106–109, 2002
Muscle development in Antalis entalis (Mollusca, Scaphopoda) and its significance for scaphopod relationships
Authors: Wanninger Andreas , Haszprunar Gerhard
Published: October 2002

We applied fluorescence staining of F‐actin, confocal laser scanning microscopy, as well as bright‐field light microscopy, SEM, and TEM to examine myogenesis in larval and early juvenile stages of the tusk‐shell, Antalis entalis. Myogenesis follows a strict bilaterally symmetrical pattern without special larval muscle systems. The paired cephalic and foot retractors appear synchronously in the early trochophore‐like larva. In late larvae, both retractors form additional fibers that project into the anterior region, thus enabling retraction of the larval prototroch. These fibers, together with the prototroch, disappear during metamorphosis. The anlagen of the putative foot musculature, mantle retractors, and buccal musculature are formed in late larval stages. The cephalic captacula and their musculature are of postmetamorphic origin. Development of the foot musculature is dramatically pronounced after metamorphosis and results in a dense muscular grid consisting of outer ring, intermediate diagonal, and inner longitudinal fibers. This is in accordance with the proposed function of the foot as a burrowing organ based on muscle‐antagonistic activity. The existence of a distinct pair of cephalic retractors, which is also found in basal gastropods and cephalopods, as well as new data on scaphopod shell morphogenesis and recent cladistic analyses, indicate that the Scaphopoda may be more closely related to the Gastropoda and Cephalopoda than to the Bivalvia. J. Morphol. 254:53–64, 2002. © 2002 Wiley‐Liss, Inc.
Organization of the nervous system in the pygmy cuttlefish, Idiosepius paradoxus ortmann (Idiosepiidae, cephalopoda)
Authors: Shigeno Shuichi , Yamamoto Masamichi
Published: October 2002

The idiosepiid cuttlefish is a suitable organism for behavioral, genetic, and developmental studies. As morphological bases for these studies, organization of the nervous system was examined in Idiosepius paradoxus Ortmann, 1881, using Cajal's silver technique and immunohistochemical staining with anti‐acetylated α‐tubulin antibody. The nervous architecture is generally identical to that described in Sepia and Loligo, but some features characterize the idiosepiid nervous system. The olfactory system is highly developed in the optic tract region. The dorsolateral lobes show large neuropils, connected with each other by a novel well‐fasciculated commissure. Each olfactory lobe is subdivided into two lobules. The neuropils of the anterior and the posterior chromatophore lobes are very poorly developed. Neuronal gigantism is not extensive in the brain; enlarged neuronal cells are visible only in the perikaryal layer of the posterior subesophageal mass. The giant nerve fiber system is of the Sepia type; the axons are not markedly thick and the first‐order giant fibers do not fuse with each other at the chiasma. Three‐dimensional images by whole‐mount immunostaining clarified the innervation pattern in the peripheral nervous system in detail. Two commissural fibers link the left and right posterior funnel nerves ventrally and dorsally. The stellate commissure, which is absent in Sepia and Sepiola, connects the stellate ganglia with each other. A branch of the visceral nerve innervating the median pallial adductor muscle is characteristically thick. Tubulinergic reactivity of the cilia and axons reveals the presence of many ciliated cells giving off an axon toward brain nerves in the surface of the funnel, head integument, arm tips, and epidermal lines. Some of these features seem to reflect the inactive nekto‐benthic life of the idiosepiid cuttlefish in the eelgrass bed. J. Morphol. 254:65–80, 2002. © 2002 Wiley‐Liss, Inc.
New thin wire formulation for time‐domain differential‐equation models
Authors: K. Choong Y. , Sewell P. , Christopoulos C.
Published: October 2002

A novel thin wire formulation for the description of thin wires in coarse numerical models is described. This new approach uses the exact local field expansions around the thin wire which are mapped onto the coarse mesh. The method is accurate, overcomes existing limitations and approximations, does not incur additional computational overheads, requires no empirical factors, and can be extended to other fine feature objects.

In this paper, the complete derivation is presented with numerical results and examples used for validation. Further significant extensions of the work is described as a separate section to emphasize possibilities and generality of the method. Copyright © 2002 John Wiley & Sons, Ltd.
Copper/zinc superoxide dismutase overexpression promotes survival of cortical neurons exposed to neurotoxins in vitro
Authors: Borg Jacques , London Jacqueline
Published: October 2002

Oxidative stress and excitotoxicity have been implicated as triggering factors in various neurodegenerative diseases or acute neurological insults. Cu/Zn superoxide dismutase (SOD1), a potent free radical scavenging factor, may prevent the progression of such diseases. In the present study, we show that SOD1 overexpression promoted the survival of cortical neuronal cultures originating from mice carrying the human SOD1 transgene. SOD1 overexpression significantly protected against the deleterious effect of reactive oxygen species, ceramide, or N‐methyl‐D‐aspartate (NMDA). It also preserved cortical neurons against apoptosis induced by NMDA or ceramide, as revealed by a smaller increase in caspase 3 activity. SOD1 overexpression was correlated with higher SOD1 activity, and neurotoxins induced an increase in SOD1 activity in cultures from both mice. Moreover, the ratio of increase of SOD1 in cultures from nontransgenic vs. transgenic mice was similar in control cultures or following neurotoxins administration. The highest amount of neurotoxin‐induced SOD1 activity was generated by NMDA. Moreover, following exposure to hydrogen peroxide, the cytoskeletal organization was altered, as evidenced by modifications of β‐tubulin or MAP2 labelling. The fact that increased superoxide dismutase activity protected neurons suggests that appropriate control of SOD1 activity is required for neuronal survival under stressful conditions. © 2002 Wiley‐Liss, Inc.
Higher order interference effects in the anisotropic Raman spectra of HCl and DCl in liquid SF6
Authors: Padilla A. , Pérez J.
Published: October 2002

The anisotropic Raman spectra of HCl and DCl in liquid SF6 were calculated in a theoretical framework which introduces the concept of higher order interference effects to describe the non‐additivity of overlapping resonances. The contributions in the different branches are discussed comparatively and the theoretical profiles are compared with existing experimental data. Copyright © 2002 John Wiley & Sons, Ltd.
Study of carvedilol by combined Raman spectroscopy and ab initio MO calculations
Authors: Marques M. P. M. , Oliveira P. J. , Moreno A. J. M. , Batista de Carvalho L. A. E.
Published: October 2002

The novel cardioprotective drug carvedilol was studied by both Raman spectroscopy and ab initio molecular orbital methods (using the density functional theory approach). The spectra, acquired both for the solid samples and DMSO solutions as a function of pH, were assigned in view of the calculated wavenumbers and intensities, and also based on the experimental data obtained for individual compounds which comprise the molecule, namely carbazole and 1,2‐dimethoxybenzene. The pH dependence of the Raman pattern of carvedilol was studied, and the pKa value of its secondary amine group was determined (pKa = 8.25) through pH titration experiments. This kind of information is of great significance for the understanding of the biochemical role of carvedilol, which is strongly determined by the acid–base behaviour of the molecule. Copyright © 2002 John Wiley & Sons, Ltd.
Fatty acid and triacylglycerol composition and thermal behaviour of fats from seeds of Brazilian Amazonian Theobroma species
Authors: Gilabert‐Escrivá M Victoria , Gonçalves Lireny A G , Silva C Rogério S , Figueira Antonio
Published: October 2002

Raw materials for cocoa butter substitutes, replacements or equivalents depend mostly on the unsteady supply from wild stands of plants, while there is no current supply of Neotropical origin. Seed fats from Theobroma species (T cacao, T bicolor, T grandiflorum, T obovatum, T subincanum, T speciosum, T sylvestre and T microcarpum, plus the closely related species Herrania mariae) were analysed for fatty acid and triacylglycerol composition by gas and liquid chromatography respectively, for iodine value, for melting point by open capillary tube and for solid fat content (SFC) by nuclear magnetic resonance. All Theobroma species had significantly lower palmitate levels than T cacao, except for T sylvestre and T speciosum, T microcarpum presented highly unsaturated fat (C18:2), while H mariae had high levels of arachidate. Fats from T sylvestre and T speciosum had a similar iodine value to T cacao and a higher melting point. No fat from the other species presented a similar melting profile to cocoa butter. T sylvestre and T bicolor were the most similar to T cacao but had a higher SFC at human body temperature. T sylvestre and T speciosum seed fats had more POP than cocoa butter. Fats from seeds of T speciosum, T sylvestre and T bicolor could be recommended as cocoa butter substitutes, while fats from species of the section Glossopetalum could be employed in products requiring fats with a lower melting point.

© 2002 Society of Chemical Industry