bluuebunny/crossref_metadata_2025 huggingface dataset:
Method for preparing thin sections of untreated equine hoof horn for electron microscopic examination
Authors: Budras Klaus‐Dieter , Schiel Christina , Mülling Christoph Karl Wolfgang , Patan Bianca
Published: October 2002

The preparation of hard tissues such as the equine hoof horn for electron microscopic examination is very difficult. In particular the penetration of fixatives and chemicals used during fixation and embedding is a problem. The objective of this study was to find and implement an alternative method enabling the preparation of high‐quality thin sections of hoof horn and other hard tissue, which maintains the hard tissue ultrastructure and can be used for immuno‐labeling. Compared to commonly used fixation and embedding techniques, the preparation of thin sections from untreated material method saves time and material and provides equivalent ultrastructural information. Furthermore, thin sections from untreated material are significantly larger and more homogeneous, more resistant to the electron ray, as well as more suitable for sectioning. The electron microscopical pictures obtained allow a comparison to previous test results achieved with fixed and embedded material. Using the preparation of thin sections from untreated material method, fixation and embedding artifacts are avoided, providing a clearer interpretation of the electron microscopical findings. Considerable advantages are achieved by using immunohistochemical techniques with untreated horn specimens because fixation invariably decreases antigenicity. Microsc. Res. Tech. 58:114–120, 2002. © 2002 Wiley‐Liss, Inc.
Neurotrophins and their receptors in the primary olfactory neuraxis
Authors: Carter Lindsay A. , Roskams A. Jane
Published: October 2002

The primary olfactory pathway is an elegant and simple system in which to study neurogenesis and neuronal plasticity because of the simple fact that olfactory receptor neurons (ORNs) are continually generated throughout the adult lifetimes of vertebrates. Thus, neuronal birth, differentiation, survival, axon pathfinding, target recognition, synapse formation, and cell death are developmental events that can be examined in the mature olfactory epithelium (OE). Neurotrophins (nerve growth factor, brain‐derived neurotrophic factor, and neurotrophin 3, and 4/5) are a family of bioactive peptides that exert their effects by interacting with high‐ and low‐affinity receptors on the surfaces of responsive cells, and have been implicated in several stages of neuronal development throughout the central and peripheral nervous system (CNS and PNS). There has been significant interest within the olfactory community as to how these multifunctional peptides might regulate the cycle of degeneration and regeneration of olfactory receptor neurons. The focus of this review is to highlight what is known about the actions of neurotrophins in the primary olfactory pathway, and to pinpoint future directions that will enable us to further understand their role in olfactory receptor neuron development and turnover. Microsc. Res. Tech. 58:189–196, 2002. © 2002 Wiley‐Liss, Inc.
Experimental manipulation of steroid concentrations in circulation and in egg yolks of turtles
Authors: JANZEN FREDRIC J. , WILSON MATTHEW E. , TUCKER JOHN K. , FORD STEPHEN P.
Published: October 2002

Steroid hormones in egg yolks are increasingly recognized as an important component of maternal and offspring fitness in oviparous vertebrates. Yet, except for in birds, the mechanism by which females allocate these resources is poorly understood. We manipulated systemic levels of hormones in reproductively mature female red‐eared slider turtles (Trachemys scripta elegans) with silastic implants to test the hypothesis that hormones are allocated to developing follicles as a quantitative function of circulating levels in the females. Turtles exhibited similar amounts (<1 ng/ml) of circulating steroids (dihydrotestosterone, estradiol‐17β, or testosterone) in early September immediately prior to experimental manipulation. After treatment with silastic implants, circulating levels of steroids increased markedly. By the following April after hibernation, circulating levels of dihydrotestosterone had returned to preimplantation levels, but circulating levels of estradiol‐17β and testosterone in estradiol‐17β‐ and testosterone‐treated turtles, respectively, remained substantially elevated through April. Focusing on testosterone, we detected nearly six‐fold higher concentrations in yolk from mature follicles from testosterone‐treated turtles than in yolk from mature follicles from control turtles. Our results provide support for the hypothesis that concentrations of steroids in egg yolks of turtles reflect circulating concentrations of steroids during follicular development rather than the hypothesis that females selectively allocate specific amounts of steroid hormones to each egg separately. Our findings also highlight an unambiguous physiological mechanism by which nongenetic maternal effects in oviparous species can directly influence the nutritional milieu experienced by developing embryos. © 2002 Wiley‐Liss, Inc.
Maternally derived yolk hormones vary in follicles of the painted turtle, Chrysemys picta
Authors: BOWDEN RACHEL M. , EWERT MICHAEL A. , FREEDBERG STEVEN , NELSON CRAIG E.
Published: October 2002

The transfer of hormones from a female to her offspring is known to occur in egg laying vertebrates, and the potential for these early, maternally derived hormones to influence sex determination in reptiles with temperature‐dependent sex determination is intriguing. In the present study, we examine variation in the concentrations of progesterone, testosterone, and estradiol among three follicle size classes within a female painted turtle (Chrysemys picta) and among females across four periods that span the pre‐ to post‐nesting season. Females were collected, and both follicles and shelled eggs (when present) were harvested for hormone analysis. Progesterone levels did not vary seasonally. However, the concentration of progesterone did vary among and within follicle classes, and was primarily dependent upon ovulatory state: Recently ovulated follicles (as yolks within shelled eggs) contained significantly more progesterone than unovulated follicles. Concentrations of testosterone were low and did not vary either among size classes or across the season. Estradiol levels decreased with increasing follicle size and were higher later in the nesting season. Thus, hormone concentrations varied among follicle sizes and states but in patterns that differed among hormones. This variation has the potential to influence sex determination. © 2002 Wiley‐Liss, Inc.
Elevated levels of trimethylamine oxide in deep‐sea fish: evidence for synthesis and intertissue physiological importance
Authors: TREBERG JASON R. , DRIEDZIC WILLIAM R.
Published: October 2002

Tissue levels of trimethylamine oxide (TMAO) were compared for seven teleost and two elasmobranch species captured from three depth ranges: shallow (<150 m), moderate (500–700 m), and deep (1,000–1,500 m). Within the teleosts, the deep‐caught species had significantly greater TMAO content than shallow‐ or moderate‐caught species. In all teleosts, muscle had substantially more TMAO than all other tissues. Kidney or, in some cases, liver had elevated trimethylamine (TMA) content, 2.20–9.65 mmol/kg, along with appreciable trimethylamine oxidase (TMAoxi) activity, suggesting active TMAO synthesis. No correlation was found between TMAoxi activity and TMAO content. The elasmobranchs in this study, Squalus acanthias and Centroscyllium fabricii from shallow and deep water, respectively, were both squaliform sharks. The deep‐caught species had significantly more TMAO in all tissues than the shallow species. Furthermore, urea was significantly less in the deep species in all tissues except liver, while the urea:TMAO ratio was significantly less in all tissues. As with teleosts, the TMAO content of muscle was substantially higher for both elasmobranchs than in all other tissues. TMAoxi was below levels of detection in both elasmobranch species, suggesting that TMAO is obtained solely from the diet. This study expands the trend of increased muscle TMAO in deep‐sea fish to a variety of other tissues. The accumulation of TMAO in various tissues in deep‐sea teleosts and the accumulation of TMAO and concurrent urea decrease in a deep‐sea elasmobranch in comparison to a shallow water species strongly support the contention that TMAO is of physiological importance in deep‐sea fish. © 2002 Wiley‐Liss, Inc.
Aperture pattern ontogeny in angiosperms
Authors: Ressayre Adrienne , Godelle Bernard , Raquin Christian , Gouyon Pierre Henri
Published: October 2002

Pollen grains display a wide range of variation in aperture number and arrangement (pattern) in angiosperms. Apertures are well‐defined areas of the pollen wall surface that permit pollen tube germination. For low aperture numbers, aperture patterns are characteristic of the major taxonomic divisions of angiosperms. This paper presents a developmental model that explains most of the aperture patterns that are recorded in angiosperms. It is based on the analysis of the different events that occur during meiosis and lead to microspore differentiation. It demonstrates that variation occurring during meiosis in angiosperms is sufficient to produce the core morphological set of the most commonly observed pollen morphologies. J. Exp. Zool. (Mol. Dev. Evol.) 294:122–135, 2002. © 2002 Wiley‐Liss, Inc.
Separation index of a graph
Authors: Vince Andrew
Published: October 2002

The concepts of separation index of a graph and of a surface are introduced. We prove that the separation index of the sphere is 3. Also the separation index of any graph faithfully embedded in a surface of genus g is bounded by a funtion of g. © 2002 Wiley Periodicals, Inc. J Graph Theory 41: 53–61, 2002
Aspects of outreach: a framework for discussion of the social benefits of microfinance
Authors: Schreiner Mark
Published: October 2002

Wide agreement about the goal of microfinance—to improve the welfare of the poor—has not led to wide agreement about how best to achieve that goal. To aid discussion, I propose a framework for outreach—the social benefits of microfinance—in terms of six aspects: worth, cost, depth, breadth, length, and scope. The framework encompasses both the poverty approach to microfinance and the self‐sustainability approach. The poverty approach assumes that great depth of outreach can compensate for narrow breadth, short length, and limited scope. The self‐sustainability approach assumes that wide breadth, long length, and ample scope can compensate for shallow depth. I show how to use the framework for BancoSol of Bolivia. Copyright © 2002 John Wiley & Sons, Ltd.
Synthesis of [18F]‐labeled 2′‐deoxy‐2′‐fluoro‐5‐methyl‐1‐β‐D‐arabinofuranosyluracil ([18F]‐FMAU)
Authors: Alauddin Mian M. , Conti Peter S. , Fissekis John D.
Published: October 2002

Synthesis of 2′‐deoxy‐2′‐[18F]fluoro‐5‐methyl‐1‐β‐D‐arabinofuranosyluracil ([18F]‐FMAU) is reported. 2‐Deoxy‐2‐[18F]fluoro‐1,3,5‐tri‐O‐benzoyl‐α‐D‐arabinofuranose 2 was prepared by the reaction of the respective triflate 1 with tetrabutylammonium[18F]fluoride. The fluorosugar 2 was converted to its 1‐bromo‐derivative 3 and coupled with protected thymine 4. The crude product mixture (5a and 5b) was hydrolyzed in base and purified by HPLC to obtain the radiolabeled FMAU 6a. The radiochemical yield of 6a was 20–30% decay corrected (d.c.) in four steps with an average of 25% in four runs. Radiochemical purity was >99% and average specific activity was 2300 mCi/μmol at the end of synthesis (EOS). The synthesis time was 3.5–4.0 h from the end of bombardment (EOB). Copyright © 2002 John Wiley & Sons, Ltd.
Identification of epitope‐like consensus motifs using mRNA display
Authors: Baggio Rick , Burgstaller Petra , Hale Stephen P. , Putney Andrew R. , Lane Meghan , Lipovsek Dasa , Wright Martin C. , Roberts Richard W. , Liu Rihe , Szostak Jack W. , Wagner Richard W.
Published: October 2002

The mRNA display approach to in vitro protein selection is based upon the puromycin‐mediated formation of a covalent bond between an mRNA and its gene product. This technique can be used to identify peptide sequences involved in macromolecular recognition, including those identical or homologous to natural ligand epitopes. To demonstrate this approach, we determined the peptide sequences recognized by the trypsin active site, and by the anti‐c‐Myc antibody, 9E10. Here we describe the use of two peptide libraries of different diversities, one a constrained library based on the trypsin inhibitor EETI‐II, where only the six residues in the first loop were randomized (6.4 × 107 possible sequences, 6.0 × 1011 sequences in the library), the other a linear‐peptide library with 27 randomized amino acids (1.3 × 1035 possible sequences, 2 × 1013 sequences in the library). The constrained library was screened against the natural target of wild‐type EETI, bovine trypsin, and the linear library was screened against the anti‐c‐myc antibody, 9E10. The analysis of selected sequences revealed minimal consensus sequences of PR(I,L,V)L for the first loop of EETI‐II and LISE for the 9E10 epitope. The wild‐type sequences, PRILMR for the first loop of EETI‐II and QKLISE for the 9E10 epitope, were selected with the highest frequency, and in each case the complete wild‐type epitope was selected from the library. Copyright © 2002 John Wiley & Sons, Ltd.
Study of laser ablation in the in vivo rabbit brain with MR thermometry
Authors: Chen Lili , Wansapura Janaka P. , Heit Gary , Butts Kim
Published: October 2002

Purpose

To investigate the peak temperature and thermal dose (T43) as tissue damage indicators for thermal therapy.

Materials and Methods

The proton resonant frequency (PRF) shift thermal coefficient was calibrated on six in vivo rabbit brains during interstitial laser ablation. The peak temperature and T43 were correlated with the lesion boundary observed on T2‐weighted spin‐echo (SE) MRI at 4 hours post‐heating in seven thermal lesions using direct MR measurement and analysis based on a binary discriminate model.

Results

The peak temperature and T43 were 48.3 ± 1.7°C and 191 ± 219 minutes, respectively, from the direct MR measurement. The values derived by the binary discriminate analysis were 47.8 ± 2.2°C and 28 ± 41 minutes, respectively.

Conclusion

Our results suggest that tissue damage in rabbit brain 4 hours after thermal ablation can be predicted reliably from a threshold temperature of approximately 48°C. J. Magn. Reson. Imaging 2002;16:147–152. © 2002 Wiley‐Liss, Inc.
Quantitation of viral load in neonatal herpes simplex virus infection and comparison between type 1 and type 2
Authors: Kimura Hiroshi , Ito Yoshinori , Futamura Masahide , Ando Yoshihiro , Yabuta Yumi , Hoshino Yo , Nishiyama Yukihiro , Morishima Tsuneo
Published: October 2002

Neonatal herpes simplex virus (HSV) infection is a severe disease with high mortality and morbidity in spite of the development of effective anti‐viral therapies. The viral load in neonatal herpes simplex virus (HSV) infection was measured retrospectively in 37 patients. HSV DNA copy numbers in serum and cerebrospinal fluid (CSF) were quantified using a real‐time PCR assay. Patients with disseminated infection had a higher viral load in their sera. whereas patients with central nervous system (CNS) infection exhibited a higher viral load in the CSF. The viral load was significantly higher in the serum of patients who died later. Interestingly, patients with HSV type‐2 infection exhibited more CNS involvement and neurological impairment, together with a high viral load in the CSF, than did HSV type‐1 patients. These results suggest that quantitation of HSV viral load may be useful for assessing the prognosis, and may provide additional information for the management of neonatal HSV infection. J. Med Virol. 67:349–353, 2002. © 2002 Wiley‐Liss, Inc.
Genotypes and S‐gene variability of Mexican hepatitis B virus strains
Authors: Sánchez L.V. , Maldonado M. , Bastidas‐Ramírez B.E. , Norder H. , Panduro A.
Published: October 2002

The genotypes and subtypes of 15 Mexican hepatitis B virus strains were determined by sequencing and phylogenetic analysis of the small S‐gene. The most predominant strains were found to be divergent genotype/subtype F/adw4 strains (66.6%), followed by A/adw2 (20.0%), D/ayw3 (6.7%), and G/adw2 (6.7%). The S‐genes of the Mexican genotype F strains and two Nicaraguan strains described previously formed a subcluster with more than 4% divergence from the other strains within this genotype. The Mexican strains within genotypes A and D showed the highest homology with strains from Europe and the United States. Ten amino acid substitutions not described previously were found in the S‐genes of strains from nine chronic carriers, whereas the S gene in strains from six acute hepatitis B patients were highly conserved as compared to their respective genotypes. One genotype F strain from an HBsAg positive chronic carrier had a T to A mutation at position 647, forming a translational stop at codon 216. Two genotype F strains from HBsAg negative chronic carriers had a Val180 instead of an Ala found in the other genotype F strains. This study shows that a divergent genotype F predominates in Mexican strains analyzed, which presented amino acid substitutions not reported previously outside the a determinant. J. Med. Virol. 68:24–32, 2002. © 2002 Wiley‐Liss, Inc.
Large‐scale comparison of experimental adjuvants with herpes simplex virus vaccine reveals a correlation of protection with IgG2a and IgG2b responses
Authors: Simms John R. , Jennings Roy , Richardson Vernon J. , Heath Andrew W.
Published: October 2002

The potential of a large number of commercial and experimental adjuvant preparations to enhance the immunogenicity of an HSV‐1 glycoprotein subunit vaccine was investigated. Evaluation was based on toxicity, HSV‐specific antibody production, and protection against lethal challenge. All adjuvants tested increased the titer of antigen specific Ig levels when compared to subunit vaccine alone. However, following challenge, a broad range of protective responses were noted. Statistically significant correlations were observed between IgG antibody levels post immunization and the observed protection and these were particularly associated with antibodies of the IgG2a and IgG2b subclasses. The results emphasize the requirement of adjuvants for vaccine formulation when using subunit preparations, and demonstrate that the magnitude and efficacy of the induced immune response varies greatly with the choice of adjuvant. J. Med. Virol. 68:82–91, 2002. © 2002 Wiley‐Liss, Inc.
Genotype spectrum of cervical human papillomavirus infection among sexually transmitted disease clinic patients in Hong Kong
Authors: Chan Paul K.S. , Mak K.H. , Cheung Jo L.K. , Tang Nelson L.S. , Chan Denise P.C. , Lo K.K. , Cheng Augustine F.
Published: October 2002

Previous studies have suggested that a few uncommon human papillomavirus (HPV) genotypes are prevalent in Chinese cervical cancer patients. To elucidate the genotype spectrum of HPVs circulating among Hong Kong Chinese, a cross‐sectional study was conducted on 553 women who attended a public sexually transmitted disease clinic. HPV DNA was detected from cervical samples using the polymerase chain reaction, followed by genotype identification based on restriction fragment length polymorphism and direct sequencing. The prevalence of HPV was 30.6% for all types combined, 14.8% for high‐risk types, 10.8% for low‐risk types, and 7.1% for unknown‐risk types. Among the HPV‐positive women, 89.9% had single type infections; whereas the other 10.1% harboured more than one HPV type. HPV11 was the most prevalent genotype, detected in 5.1% of subjects; followed by HPV16 (4.9%), HPV58 (4.3%), HPV6 (3.3%), and HPV53 and CP8304 (2.2% each). Other less common genotypes found were HPV18, 33, 39, 61, LVX160, MM4, MM7 (range: 0.7–1.6%); HPV26, 45, 54, 56, 59, and LVX100 (range: 0.4–0.5%); HPV35, 40, 52, 55, 68, MM8, and MM9 (0.2% each). This study shows that HPV58 is the second most common high‐risk HPV genotype circulating among Chinese female sexually transmitted disease clinic patients in Hong Kong. This together with previous reports of the high prevalence of HPV58 among Chinese cervical cancer patients accentuate the importance of developing vaccines targeting at this otherwise uncommon genotype. J. Med. Virol. 68: 273–277, 2002. © 2002 Wiley‐Liss, Inc.
Unusual manifestations of primary cytomegalovirus infection in patients without HIV infection and without organ transplants
Authors: Crowley Brendan , Dempsey Jed , Olujohungbe Ade , Khan Abdul , Mutton Ken , Hart C.A.
Published: October 2002

Primary cytomegalovirus (CMV) infection with marked constitutional symptoms is rare in immunocompetent individuals and in those with iatrogenic immunosuppression, except transplant recipients. Four patients admitted to hospital with clinical illnesses associated with primary CMV infection were identified over a 12‐month period. Their medical records were reviewed with regard to clinical and laboratory data, and outcome. Primary CMV infection was defined by the concomitant presence of CMV IgM and low avidity CMV IgG antibody. Of two patients with no known underlying illness, one presented with thrombocytopenic purpura and the other with vasculitis. Two patients receiving immunosuppressants for underlying ulcerative colitis presented with CMV‐induced pancytopenia and CMV colitis. Atypical lymphocytosis was a feature on blood film examination in three of the four cases. One patient with disseminated CMV infection died of progressive multiorgan failure despite antiviral treatment. CMV disease following primary CMV infection should be considered in otherwise immunocompetent individuals with atypical lymphocytosis on blood film analysis, and particularly in patients on immunosuppressants such as those with ulcerative colitis, since early diagnosis and treatment with antiviral drugs may improve outcome. J. Med. Virol. 68: 237–240, 2002. © 2002 Wiley‐Liss, Inc.
Electromagnetic field representations and computations in complex structures III: network representations of the connection and subdomain circuits
Authors: Russer Peter , Mongiardo Mauro , Felsen Leopold B.
Published: October 2002

When complex structures are divided into subdomains for electromagnetic field computations, it is necessary to specify which of the tangential electromagnetic field components at the boundaries may be regarded as independent and which have to be treated as dependent. In this third of our three‐part study, we establish some rules for the choice of primary and secondary fields at the connection interface, i.e. at the interface between adjacent subdomains. The discretized field continuity equations at the connection interface provide the connection network whose canonical forms are illustrated, and it is shown that the normalized scattering matrix is symmetric, orthogonal and unitary.

Tellegen's theorem is introduced in order to provide the basis for consistent choice of primary and secondary fields and for deriving canonical forms of the connection network. The field problem is systematically treated by partitioning and by specifying canonical Foster representations for the subcircuits. Connection between different subdomains is obtained by selecting the appropriate independent field quantities via Tellegen's theorem. For each subdomain, as well as for the entire circuit, an equivalent circuit extraction procedure is feasible, either in closed form for subdomains amenable to analytical treatment or via the relevant pole structure description when a numerical solutions is available. Moreover, circuit‐based resonant expansions in the complex frequency plane are provided here for the analytic dyadic Green's functions in [1]. Copyright © 2002 John Wiley & Sons, Ltd.
Neuronal nitric oxide synthase modulates basal catecholamine secretion in bovine chromaffin cells
Authors: Vicente S. , González M.P. , Oset‐Gasque M.J.
Published: October 2002

The role of endogenously produced nitric oxide (NO) in the regulation of basal catecholamine (CA) secretion was studied in chromaffin cells. Treatment of chromaffin cells with nitric oxide synthase (NOS) inhibitors produced a dose‐dependent increase in basal catecholamine secretion, which paralleled their ability to inhibit NOS activity. This inhibitory profile was similar to that found in neurons, suggesting the constitutive expression of neuronal NOS (nNOS) in these cells, which was confirmed by Western blot analysis. A study of the kinetics and pharmacology of nNOS activity expressed in chromaffin cells in culture indicated that NOS activity is calcium‐dependent, increases with time, and is highly dependent on both intracellular concentrations of L‐arginine (Km ∼ 4 μM, Vmax = 908 ± 60 pmol/hr × 106 cells) and transport of L‐arginine into the cells (exhibiting two affinity constants of k1 = 3.2 ± 0.3 μM and k2 = 126 ± 5.5 μM). The effects of NOS inhibitors on CA secretion were mediated by the L‐arginine‐NO‐cGMP pathway, insofar as exogenous L‐arginine was able to partially block the increase in CA secretion evoked by them, and 1H‐[1,2,4]oxadiazolo[4,3‐a]quinoxaline‐1‐one (ODQ), a specific inhibitor of guanylate cyclase, and zaprinast, an inhibitor of the cGMP phosphodiesterase, were able to increase and inhibit, respectively, basal CA secretion in a dose‐dependent manner. These results suggest that chromaffin cells exhibit a tonic production of NO by nNOS that keeps the basal CA secretion at low levels, and this could be necessary for maintaining a normotensive state. © 2002 Wiley‐Liss, Inc.
Striatal neurochemical changes in transgenic models of Huntington's disease
Authors: Ariano Marjorie A. , Aronin Neil , Difiglia Marian , Tagle Danilo A. , Sibley David R. , Leavitt Blair R. , Hayden Michael R. , Levine Michael S.
Published: October 2002

Transgenic mouse models of Huntington's disease (HD) were examined following the onset of overt behavioral symptoms. The HD transgenic mice demonstrated profound striatal losses in D1, D2, and D3 dopamine (DA) receptor proteins in comparison with their nonsymptomatic, age‐matched littermate controls. In parallel, a robust increase in the striatal D5 DA receptor subtype occurred in the transgenic compared with the wild‐type control mice. This receptor elevation was accompanied by heightened cyclic AMP levels, which may be induced by the adenylyl cyclase‐linked D5 receptor. This is a unique result; normal striatal D5 protein levels are modest and not thought to contribute substantially to cyclic AMP‐mediated DA signaling mechanisms. Simple compensatory up‐regulation of D5 DA receptors in response to D1 receptor subtype loss does not explain our findings, because genetic inactivation of the D1 DA receptor does not alter levels of D5 DA receptor expression. Immunofluorescent detection of tyrosine hydroxylase showed that nigrostriatal DA containing terminals were reduced, further supporting that disturbances in DA signaling occurred in HD transgenic models. The substance P‐containing striatal efferent pathway was more resistant to the HD mutation than met‐enkephalin‐producing striatal projection neurons in the transgenics, based on neuropeptide immunofluorescent staining. Analogous findings in multiple transgenic models suggest that these changes are due to the presence of the transgene and are not dependent on its composition, promotor elements, or mouse strain background. These findings suggest modifications in the striatal DA system and that its downstream signaling through cyclic AMP mechanisms is disrupted severely in HD following onset of motor symptoms. © 2002 Wiley‐Liss, Inc.
Molecular analysis of the vagal motoneuronal degeneration after right vagotomy
Authors: Ji Junfeng , Dheen S. Thameem , Tay Samuel Sam Wah
Published: October 2002

The aim of this study was to investigate the vagal motoneuronal degeneration after right vagotomy using in situ hybridization, RT‐PCR, and immunohistochemistry methods. The morphology of the vagal motoneurons in dorsal motor nucleus of the vagus nerve (DMV) and nucleus of ambiguus (NA) after right vagotomy was examined by using Nissl staing and TUNEL. The expression of inducible nitric oxide synthase (iNOS), bcl‐2, bax, and caspase‐3 in DMV and NA of rats after right vagotomy was studied. Additionally, the involvement of the N‐methyl‐D‐aspartate (NMDA) receptor‐calcium‐neuronal nitric oxide synthase (nNOS) pathway in the vagal motoneuronal degeneration was addressed by double‐immunolabeling analysis of nNOS with NMDAR1 and calbindin D28K in right‐vagotomized rats. The neurons in right DMV and NA displayed a darkly stained, shrunken morphology at 1 day and 5 days following right vagotomy as shown by Nissl staining. Quantitative analysis revealed that, at 1 day and 5 days following right vagotomy, the number of neurons in right DMV, but not NA, was significantly reduced in comparison with that of control rats. Occasional TUNEL‐positive neurons were detected in right DMV of rat at 1 day after right vagotomy. The expression of iNOS protein and mRNA was absent in DMV and NA of control rats. However, the iNOS mRNA expression was induced bilaterally in DMV and NA at 1 day postoperation and continued to be up‐regulated until 5 days after vagotomy as shown by in situ hybridization. Immunohistochemistry analysis also showed the increased expression of iNOS in bilateral DMV and NA of vagotomized rats. RT‐PCR analysis revealed the enhanced bcl‐2 and reduced bax mRNA levels and subsequent up‐regulation of both bcl‐2 and bax mRNA in right sides of the vagotomized brainstems at 1 day and 5 days postoperation, respectively. In situ hybridization analysis confirmed the up‐regulation of bcl‐2 and bax mRNA in right DMV and NA of the rats at 5 days following operation. Immunohistochemistry analysis showed up‐regulated Bcl‐2 immunoreactivity and undetectable changes in Bax immunoreactivity in DMV and NA of rats at 1 day after vagotomy, whereas enhancement of both Bcl‐2 and Bax immunoreactivity was observed at 5 days postoperation. In addition, the caspase‐3 mRNA level was elevated ipsilaterally in DMV and NA at 1 day and 5 days following right vagotomy. Double‐immunofluorescence analysis showed complete colocalization of nNOS with NMDAR1 and with calbindin in ipsilateral DMV and NA at 10 days following right vagotomy. This study suggests that the signal pathway for NMDAR1‐calcium‐nNOS and the up‐regulation of iNOS in DMV and NA may be involved in the vagal motor neurodgeneration after right vagotomy. Furthermore, our results imply that the apoptosis pathway mediated by Bcl‐2, Bax, and caspase‐3 may be activated in vagal motoneurons after right vagotomy. © 2002 Wiley‐Liss, Inc.
Enhanced resolution modelling study on anthropogenic climate change: changes in extremes of the hydrological cycle
Authors: Voss Reinhard , May Wilhelm , Roeckner Erich
Published: October 2002

Changes in variability and extremes of the hydrological cycle are studied in two 30 year simulations using a general circulation model at high horizontal resolution. The simulations represent the present‐day climate and a period in which the radiative forcing corresponds to a doubling of the present‐day concentrations of atmospheric greenhouse gases. In most regions and seasons the probability density function of daily precipitation experiences a stretching associated with a higher probability of heavy precipitation events in the warmer climate. Whereas extremely long wet spells show only moderate changes, the extremely long dry spells are extended at middle latitudes over most land areas. At high latitudes the changes in annual maximum river runoff are mainly controlled by changes in snow budget. Eight out of 14 selected major rivers show a statistically significant change in 10 year return values of the annual maximum discharge. In two cases a significant decrease is found and in six cases there is a significant increase. Copyright © 2002 Royal Meteorological Society
Decontamination of aflatoxin B1‐contaminated corn by ammonium persulphate during fermentation
Authors: Burgos‐Hernández Armando , Price Ralph L , Jorgensen‐Kornman Karen , López‐García Rebeca , Njapau Henry , Park Douglas L
Published: October 2002

The decontamination of aflatoxin B1(AFB1)‐contaminated corn, which is required if the corn is to be suitable for alternative use, by an ammoniation–fermentation integrated process was studied. This process could be used for the production of fuel ethanol from aflatoxin‐containing corn. Different concentrations (0.25, 0.50, 1.00, 1.5 and 2.0% w/w) of ammonium persulphate were tested in the detoxification of AFB1‐contaminated corn during fermentation. In order to increase the decontamination of corn, 0.5 and 1.0% (w/w) azodicarbonamide, benzoyl peroxide and hydrogen peroxide were tested. Peroxides were added at three different stages of the fermentation process: liquefaction, saccharification and fermentation. Levels of AFB1 and ethanol were determined after each fermentation process. Treated corn was tested for mutagenic potential using the Ames test with TA100 tester strain and pure AFB1 as positive control. Addition of 2.0% (w/w) ammonium persulphate caused the highest level of decontamination without affecting ethanol production. Addition of peroxides did not significantly (P < 0.05) increase ethanol production or significantly (P < 0.05) improve the decontamination process. The best processes for decontamination of corn and for ethanol production included the addition of 2.0% (w/w) ammonium persulphate for both and of 1.0 and 0.5% (w/w) benzoyl peroxide respectively. All treated corn samples showed no mutagenic potential. Possible industrial use of these processes is discussed.

© 2002 Society of Chemical Industry
Effect of feeding oilseed supplements to dairy cows on ruminal and milk fatty acid composition
Authors: McNamee Brian F , Fearon Ann M , Pearce Jack
Published: October 2002

The objective of this study was to compare the effects of oilseed‐based supplements, rapeseed and linseed, against a barley‐based control, on the fatty acid composition, and subsequent solid fat ratio, of the milk fat from dairy cows. In addition, as a means of understanding the digestive processes which influence the milk fat composition, ruminal extracts were collected from the cows and analysed for fatty acid composition. Four lactating dairy cows each fitted with a rumen fistula were provided with silage and one of four concentrate diets. The main constituent of the concentrate supplements was either rapeseed (ground or unground), linseed (unground) or a barley control. The diets were offered in accordance with a 4 × 4 Latin square arrangement. The oilseed‐supplemented concentrates provided the cows with 620–640 g fatty acids day−1. Experimental treatments were provided to the cows for 2 weeks, after which ruminal extracts were collected over a 24 h period and a milk sample was taken. All extracts were analysed for fatty acid composition. The diets fed influenced the long‐chain fatty acid composition of the ruminal extracts and milk fat. The proportion of C18:1n‐9 in the ruminal extracts increased from 202–224 to 282–321 g kg−1 of the total fatty acids when the cows were provided with the rapeseed‐based diets. The linseed‐based diet increased the C18:1n‐9 proportion of the ruminal extracts from 164 to 218 g kg−1 of the total fatty acids. Both rapeseed‐based diets also resulted in a higher proportion of C18:0 in the ruminal extract, possibly owing to biohydrogenation of the dietary fatty acids. This proportion of C18:0 in the ruminal extract was lowest immediately after feeding, increasing to a maximum 4–6 h later. Both rapeseed‐based concentrates increased the proportion of C18:1n‐9 in the milk fat to approximately 300 g kg−1 of the total fatty acids as compared with 214 g kg−1 for the control. The proportion of C18:1n‐9 in the milk fat from the cows offered the linseed‐based concentrate was 246 g kg−1 of the total fatty acids. There were also significant decreases in the proportions of C16:0 in the milk fat from the cows offered all oilseed‐based concentrates. There was no difference between the fatty acid compositions of the milk fats from the cows fed the ground or unground rapeseed‐based supplements. The oilseed‐based supplements also resulted in significant decreases in the solid fat content of the milk fat at temperatures ranging from 0 to 35 °C, which would be indicative of a softer, more spreadable butter.

© 2002 Society of Chemical Industry
Profiles of volatile compounds and sensory analysis of three blends of coffee: influence of different proportions of Arabica and Robusta and influence of roasting coffee with sugar
Authors: Sanz Cristina , Maeztu Laura , Zapelena Ma Jose , Bello José , Cid Concepción
Published: October 2002

One hundred and forty‐six volatile compounds were identified and quantified using a static headspace sampler in three blends of coffee: Arabica/Robusta 80:20 (A80:R20) natural roasted coffee, Arabica/Robusta 20:80 (A20:R80) natural roasted coffee and Arabica/Robusta 20:80 with 50% of Robusta coffee roasted with sugar (A20:R80 50% Torrefacto). The different proportion of Arabica and Robusta coffee in the blend A80:R20 versus A20:R80 influenced the amounts of 20 chemical families of volatile compounds. Aldehydes, ketones, alcohols, pyrroles, pyrazines, furans, thiazoles, thiophenes, esters, oxazoles, lactones, sulphur compounds, pyridines, alkanes, alkenes, phenolic compounds, benzenic compounds, acids, pyranones and terpenes were present in higher quantities in the sample containing 80% of Arabica coffee, whereas sulphur compounds were more abundant in the coffee with 80% of Robusta. Sensory differences were also found between the two blends of coffee in the burnt, caramel, nutty, earthy and roasty notes. Torrefacto coffee, widely consumed in Spain, is obtained by roasting coffee with sugar. Higher quantities of ketones, alcohols, pyrazines, furans, pyridines, alkanes, phenolic compounds, pyranones and terpenes were found in the blend A20:R80 50% Torrefacto coffee versus A20:R80 natural roasted coffee. These differences in the volatile fraction were perceived by our panellists in the intensities of the nutty, roasty, earthy, burnt and caramel notes.

© 2002 Society of Chemical Industry
Methods for the preparation of cell walls from potatoes
Authors: Jardine W G , Doeswijk‐Voragen C H L , MacKinnon I M R , van den Broek L A M , Ha M A , Jarvis M C , Voragen A G J
Published: October 2002

A group of new methods is described for preparing cell walls from potatoes and processed potato products. Starting from raw domestic potatoes, starch is degraded enzymatically after a very brief 100 °C gelatinisation step conducted after homogenisation to minimise the time required for heat transfer. Protein is removed by detergent and phenol extraction. This procedure (method 1) gave cell wall preparations containing <5% starch, with minimal degradation of wall polysaccharides. It did not, however, remove starch efficiently from industrial potatoes in which the starch content is much higher. A different procedure, method 2, was used in this case. In method 2 a 20 min starch gelatinisation step was used but the temperature was restricted to 70 °C and the pH to 4.0, with the aim of protecting pectins from depolymerisation. Method 2 and method 1A, which is a hybrid procedure involving the starch gelatinisation step from method 2 and other steps from method 1, gave low‐starch cell walls from industrial as well as domestic potatoes. These methods are suitable for a range of potato types and potato products and are either more efficient or more convenient than previous procedures for cell wall isolation.

© 2002 Society of Chemical Industry
Transfer of heat and moisture during oven baking of potatoes
Authors: Wilson W D , MacKinnon I M , Jarvis M C
Published: October 2002

Potatoes were baked for up to 60 min in a conventional fan‐assisted oven. Temperature profiles within the potato tubers were determined both by direct measurement and by following the inward progress of starch gelatinisation, which occurs at 65 °C. Temperature profiles with time were S‐shaped and about 30 min was needed for the centre to reach 100 °C. From the nature of the temperature profiles, long potato tubers will cook faster than round tubers of the same weight, and a varietal ‘shape factor’ was defined to quantify this effect. The slow temperature rise, compared to other forms of cooking, was due to evaporative cooling. Moisture loss was linear with time after an initial lag and reached 15–20% of the mass of the potato after 1 h of cooking. Approximately half of the moisture was lost from the outermost layer of the potato under the periderm, leaving behind a dried layer of flesh that appeared to restrict water transport and, unlike any other part of the potato, could exceed 100 °C. This has consequences for the thermal development of flavour compounds as well as for perceived texture.

© 2002 Society of Chemical Industry
Loss of ascorbic acid during storage of Conference pears in relation to the appearance of brown heart
Authors: Eccher Zerbini Paola , Rizzolo Anna , Brambilla Ada , Cambiaghi Paola , Grassi Maurizio
Published: October 2002

To investigate the causes of brown heart (BH) in Conference pears, the contents of ascorbic acid (AA) and dehydroascorbic acid (DHAA) in fruits were studied during the first period of storage. AA was studied in 1999 and 2000, DHAA in 1999 only. Pears from normal and very late harvest stored immediately or after a delay in a controlled atmosphere with 2% O2 and low (0.7%) or high (5%) CO2 were compared. DHAA content did not change during storage in any treatment. AA decreased in storage according to an exponential model. The rate of AA loss was different in the two years and was higher in late‐harvest fruits and in those stored in 5% CO2. BH appeared in different treatments when AA decreased below a threshold, which was about 2 mg kg−1 fresh weight, corresponding to 5% of the AA content at harvest. The frequency and severity of symptoms depended linearly on the time spent by fruits in low‐ascorbate conditions. Advanced maturity at harvest and storage in high CO2 determine the appearance of BH and increase the rate of AA loss, probably through a reduction in the turnover of antioxidants.

© 2002 Society of Chemical Industry
Relationship between free amino acids and glucosinolates in primary and secondary inflorescences of 11 broccoli (Brassica oleracea L var italica) cultivars grown in early and late seasons
Authors: Rosa Eduardo , Gomes M Helena
Published: October 2002

Amino acids have been identified as the precursors of a class of secondary plant metabolites, the glucosinolates, but there is no report on the relationship between the two in vivo. In this work we have investigated the relationship between free amino acids and glucosinolates in 11 broccoli cultivars grown in early (April–July) and late (August–January) seasons. Free amino acids and glucosinolates were quantified by HPLC in primary and secondary inflorescences at maturity stage. Analysis of the data revealed that no significant correlation exists between free amino acids and glucosinolates, suggesting that the precursors of these compounds might be used during the first step of the biosynthetic process to form intermediates, the aldoximes and thiohydroximates, which will then be used in the second stage of the biosynthesis to form, apart from glucosinolates, other secondary plant metabolites.

© 2001 Society of Chemical Industry
Sole brachytherapy of the tumor bed after conservative surgery for T1 breast cancer: Five‐year results of a phase I–II study and initial findings of a randomized phase III trial
Authors: Polgár Csaba , Sulyok Zoltán , Fodor János , Orosz Zsolt , Major Tibor , Takácsi‐Nagy Zoltán , Mangel László C. , Somogyi András , Kásler Miklós , Németh György
Published: October 2002

Background and Objectives

The objectives of this study were to test the feasibility of sole interstitial high‐dose‐rate brachytherapy (HDR‐BT) after breast‐conserving surgery (BCS) for T1 breast cancer in a phase I–II study, and to present the initial findings of a phase III trial comparing the efficacy of tumor bed radiotherapy (TBRT) alone with conventional whole breast radiotherapy (WBRT).

Methods

Forty‐five prospectively selected patients with T1 breast cancer undergoing BCS were enrolled into a phase I–II study of TBRT alone, using interstitial HDR implants. HDR‐BT of 7 × 4.33 Gy (n = 8) and 7 × 5.2 Gy (n = 37) was delivered to the tumor bed. Based on the results of this phase I–II study, a further 126 patients were randomized to receive 50 Gy WBRT (n = 63) or TBRT alone (n = 63); the latter consisted of either 7 × 5.2 Gy HDR‐BT (n = 46) or 50‐Gy wide‐field electron irradiation (n = 17). Breast cancer related events and side effects were assessed.

Results

In the phase I–II study, at a median follow‐up of 57 months, 2 (4.4%) local, 3 (6.7%) axillary, and 3 (6.7%) distant failures were observed. Two patients (4.4%) died of breast cancer. The 5‐year probability of cancer‐specific, relapse‐free and local recurrence‐free survival was 90.0%, 85.9%, and 95.6%, respectively. The cosmetic results were judged to be excellent in 44 of 45 patients (97.8%). Severe (higher than grade 2) skin sequelae or fibrosis was not found. Symptomatic fat necrosis occurred in one patient (2.2%). In the phase III study, at a median follow‐up of 30 months, the locoregional tumor control was 100% in both arms. The 3‐year probability of cancer‐specific and relapse‐free survival was 98.1% and 98.4% in the WBRT group and 100% and 94.4% in the TBRT group, respectively (P = NS). There was no significant difference between the two treatment arms regarding the incidence of radiation side effects.

Conclusions

Five‐year results of our phase I–II study prove that sole HDR‐BT of the tumor bed with careful patient selection and adequate quality assurance is a feasible alternative to WBRT. However, long‐term results of phase III trials are required to determine the equivalence of TBRT alone, compared with WBRT in the management of selected patients with early breast cancer. J. Surg. Oncol. 2002;80:121–128. © 2002 Wiley‐Liss, Inc.
Hepatic arterial infusion of 5‐fluorouracil and cisplatin for unresectable or recurrent hepatocellular carcinoma with tumor thrombus of the portal vein
Authors: Itamoto Toshiyuki , Nakahara Hideki , Tashiro Hirotaka , Haruta Naoki , Asahara Toshimasa , Naito Akira , Ito Katsuhide
Published: October 2002

Background and Objectives

This study was designed to evaluate the efficacy of hepatic arterial infusion of 5‐fluorouracil (5‐FU) and cisplatin (CDDP) for unresectable or recurrent hepatocellular carcinoma (HCC) with tumor thrombus of the trunk or first branches of the portal vein (PVTT).

Methods

Seven unresectable or recurrent HCC patients with PVTT were enrolled in this study. A one‐week course of chemotherapy consisted of intraarterial administration of CDDP (10 mg on days 1–5) for 1 h and 5‐FU (250 mg on days 1–5) for 24 h, followed by cessation of administration for the subsequent 2 days (days 6 and 7). Three or more sequential courses of chemotherapy were given through an implanted reservoir.

Results

Serum α‐fetoprotein (AFP) levels before the chemotherapy were >20 ng/ml in six of the seven patients. Serum AFP levels were decreased in four of the six patients after chemotherapy, including two patients (cases 1 and 7) whose AFP levels later returned to normal. Six of the seven patients had measurable lesions in the liver, with a response rate of 33%. In three of the seven patients (43%), PVTTs decreased in size or disappeared after chemotherapy. The mean and median survival periods of all patients were 8.0 and. 7.5 months, respectively.

Conclusions

The chemotherapy described in this report is beneficial in terms of survival for HCC patients with PVTT for whom transcatheter arterial embolization or surgical treatment is contraindicated. J. Surg. Oncol. 2002;80:143–148. © 2002 Wiley‐Liss, Inc.