bluuebunny/crossref_metadata_2025 huggingface dataset:
Methyl palmitate prevents CCl4‐induced liver fibrosis
Authors: Rodríguez‐Rivera Adriana , Galicia‐Moreno Marina , Reyes‐Gordillo Karina , Segovia José , Vergara Paula , Moreno Mario G. , Shibayama Mineko , Tsutsumi Víctor , Muriel Pablo
Published: July 2008

Liver fibrosis is characterized by an excess of collagen fiber deposition, and it is known that Kupffer cells play an important role by immunomodulation of the toxic response. Methyl palmitate (MP) is an effective Kupffer cell inhibitor. The aim of this work was to evaluate the effect of MP on experimental liver fibrosis. Four groups were formed: the control group, which received the vehicles only; CCl4 group (0.4 g kg−1, i.p., three times a week, for eight weeks); CCl4 plus MP (300 mg kg−1, i.p., daily); and MP alone. Alanine aminotransferase was increased by CCl4, and MP did not prevent this increase. Lipid peroxidation was increased markedly by CCl4; again, MP was not able to prevent this effect. Fibrosis increased nearly 6‐fold (measured as liver hydroxyproline content) in the CCl4 group; MP preserved the normal content of collagen. These results were corroborated by histopathology. To elucidate the antifibrogenic mechanism of MP, we measured the production of TGF‐β; CCl4 increased this cytokine several‐fold, and MP abolished this increase. Collectively the present results indicate that MP possesses a strong antifibrogenic effect at least in the CCl4 model of fibrosis. The antifibrotic effect of MP is probably associated with its ability to reduce TGF‐β content, maybe by immunomodulation of Kupffer cells functioning. Copyright © 2008 John Wiley & Sons, Ltd.
Application of structure–activity relationships to investigate the molecular mechanisms of hepatocyte toxicity and electrophilic reactivity of α,β‐unsaturated aldehydes
Authors: Chan Katie , Poon Raymond , O'Brien Peter J.
Published: July 2008

Covalent binding of reactive electrophiles to cellular targets is a molecular interaction that has the potential to initiate severe adverse biological effects. Therefore, a measure for electrophilic reactivity with biological nucleophiles could serve as an important correlate to toxic effects such as hepatocyte death. To determine if electrophile reactivity correlates with rat hepatocyte cytotoxicity, the inherently electrophilic α,β‐unsaturated aldehydes were chosen for investigation. Reactivity was measured with simple assays that used glutathione, a soft nucleophile, and butylamine, a harder nucleophile, as models for protein thiol and amine nucleophilic sites, respectively. Despite their higher reactivity with thiols, a linear relationship was only observed between hepatocyte cytotoxicity and amine reactivity. Structure–activity relationships were also investigated for hepatocyte toxicity, and results showed toxicity was well modelled by log P and electronic parameters ELUMO and partial charge of the carbonyl carbon (
). Hydrophobicity and electronic descriptors were only significant in separate distinct models, suggesting that there were simultaneously occurring mechanisms that affected toxicity. Log P was linked to the ease of oxidation by a microsomal aldehyde dehydrogenase enzyme, while the electronic descriptors and amine reactivity were linked to direct alkylation. Even with the presence of electrophile characteristics, α,β‐unsaturated aldehyde hepatocyte toxicity could not be predicted exclusively by electrophilic reactivity as oxidative metabolism was also a factor for toxicity. Copyright © 2008 John Wiley & Sons, Ltd.
Increased levels of urinary nitrite and nitrotyrosine in Yusho victims 40 years after accidental poisoning with polychlorinated biphenyls in Nagasaki, Japan
Authors: Shimizu Kazuhiro , Ogawa Fumihide , Thiele Jens J. , Lee Jeong Boem , Bae SangJae , Sato Shinichi
Published: July 2008

Forty years have passed since the accidental poisoning with polychlorinated biphenyls (PCB) in Japan in 1968, named Yusho. High concentrations of PCB are still detected in the serum of the Yusho victims. PCB produces superoxide (O2−) in the metabolic process and we reported high concentrations of serum nitrite, a stable metabolite reflecting nitric oxide (NO), in the Yusho victims. NO reacts with O2− and immediately produces peroxynitrite (ONOO−). ONOO− causes nitration of tyrosine residues and produces nitrotyrosine (NT). Therefore, we measured urinary concentrations of nitrite and NT in the victims and age‐matched controls. The mean urinary concentrations of nitrite and NT were significantly higher than in the controls. There was a positive correlation between urinary nitrite and NT in the Yusho victims. Furthermore, there was a positive correlation between the ratio of urinary NT to nitrite and serum PCB concentrations in the Yusho victims. It was considered that the emergence of some ailments could be presumed to have been caused by high levels of urinary nitrite and NT in the Yusho victims. Copyright © 2008 John Wiley & Sons, Ltd.
Secretion of cortisol and aldosterone as a vulnerable target for adrenal endocrine disruption — screening of 30 selected chemicals in the human H295R cell model
Authors: Ullerås Erik , Ohlsson Åsa , Oskarsson Agneta
Published: July 2008

The adrenal gland is a vulnerable target for toxic insult. Disruption of adrenal steroidogenesis and hormone secretion may cause serious effects on human health. A human in vitro model is needed to predict effects, and elucidate mechanisms of endocrine disruption and adrenal toxicity. The human adrenocortical cell line H295R has been used to screen for effects on sex hormones. Here, we have analyzed the effect of 30 potential endocrine disrupting chemicals on the secretion of cortisol and aldosterone from the H295R cells, using specific ELISA assays. The effect of chemicals was analyzed for basal and forskolin‐ or angiotensin II‐stimulated hormone secretion. The chemicals were tested at the highest concentration where they displayed no evident unspecific cytotoxicity. Quantitative and qualitative differences in effects on hormone secretion were demonstrated for the various chemicals. A subset of the chemicals displayed different effects on cortisol and aldosterone secretion, and in some cases the effects were different between basal and stimulated hormone secretion. Aminoglutethimide, prochloraz, ketoconazole, 6‐hydroxyflavone, imazalil and etomidate had the most marked inhibitory effects on cortisol (with or without forskolin) and ketoconazole, 6‐hydroxyflavone, imazalil and etomidate had the most marked effects on aldosterone (with or without angiotensin II). The results are discussed in terms of known effects, structural similarity and possible mechanisms. We have shown that adrenal steroidogenesis is a vulnerable target for toxic insult and that the H295R assay is a useful in vitro model for screening purposes. Copyright © 2008 John Wiley & Sons, Ltd.
Paratracheal lymph nodes: A new sonographic finding in autoimmune thyroiditis
Authors: Serres‐Créixams Xavier , Castells‐Fusté Ignasi , Pruna‐Comella Xavier , Yetano‐Laguna Victorina , Garriga‐Farriol Victòria , Gallardo‐Agromayor Elena
Published: July 2008

Purpose.

To assess the clinical value of paratracheal lymph nodes (PLNs) as a novel sonographic finding in autoimmune thyroiditis (AT).

Methods.

A total of 309 consecutive patients underwent sonographic examinations of the thyroid between 1998 and 2003. A single radiologist assessed the sonographic findings of AT and PLNs. All patients underwent serological tests for antimicrosomal antibodies (AMAs). Patients with clinical, cytological, or laboratory findings of thyroiditis formed the AT group with a positive AMA test (n = 199). Controls were patients with no signs of nodular thyroid disease, normal thyrotropin, negative AMA, and benign cytology (n = 110).

Results.

PLNs were seen in 184 of 199 patients in the AT group and in 28 of 110 controls (P < 0.001) (sensitivity of 93.4%, specificity of 74.5% in the diagnosis of AT). PLNs in controls were fewer (2.8 ± 1.5 versus 4.7 ± 2.6; P < 0.001) and smaller (8.2 ± 2.4 mm versus 10.7 ± 3.3 mm; P < 0.001) than in the AT group.

Conclusion.

PLNs are often present in patients with AT and are detectable with sonography. Radiologists should be aware of the importance of including the paratracheal region in the evaluation of the thyroid gland. © 2008 Wiley Periodicals, Inc. J Clin Ultrasound, 2008
Improved prenatal aneuploidy screening using the novel advanced first‐trimester screening algorithm: A multicenter study of 10,017 pregnancies
Authors: Schmidt Peter , Hörmansdörfer Cindy , Pruggmayer Michael , Schütte Christian , Neumann Armin , Gerritzen Andreas , Vaske Bernhard , Hillemanns Peter , Scharf Alexander
Published: July 2008

Purpose.

It has been postulated that the maternal age component should be completely excluded from first‐trimester screening (FTS) for fetal aneuploidies. In this study, we tested a new algorithm known as advanced first‐trimester screening (AFS), which disregards maternal age.

Method.

In a multicenter study, FTS findings were retrieved from 10,017 pregnancies. FTS risk assessment was performed using the Nicolaides method, and the AFS score was calculated. The results of both methods were compared.

Results.

Within this population, 81 fetuses had an abnormal karyotype. The sensitivity of the 2 algorithms was 86.4%. When the AFS method was used, the positive predictive value rose from 9.6% (FTS) to 12.4% (AFS). Using AFS, the test positive rate could be decreased by 161 cases (−22.2%) (p < 0.0001), due to a reduction of false positive cases. As a result, the false positive rate of AFS was 24.5% lower than that of FTS, while the same number of aneuploidies was detected.

Conclusion.

AFS can markedly reduce the rate of false positive test results. If these results are confirmed by larger multicenter studies, the new AFS will represent a great improvement in fetal aneuploidy screening. © 2008 Wiley Periodicals, Inc. J Clin Ultrasound, 2008
Extracranial and intracranial vertebral artery dissection: Long‐term clinical and duplex sonographic follow‐up
Authors: Wessels Tiemo , Mosso Maria , Krings Timo , Klötzsch Christof , Harrer Judith U.
Published: July 2008

Purpose.

To determine the value of color Doppler sonography (CDUS) in the diagnosis and follow‐up of patients with extracranial and intracranial vertebral artery (VA) dissection.

Method.

Thirty‐three patients aged 42 ± 12 years with 40 VADS confirmed via digital subtraction angiography (DSA) (n = 37) and magnetic resonance angiography (MRA) (n = 3) were included in the study. All patients were investigated with extracranial CDUS and transcranial CDUS (TCCDUS) over a mean ± SD follow‐up period of 42 ± 24 months and occurrence of new ischemic symptoms was assessed. Sonographic results were compared with initial and follow‐up angiographic results.

Results.

At presentation, 24/33 (73%) patients had suffered an ischemic stroke, 5/33 (15%) had a transient ischemic attack (TIA), and 4/33 (12%) were asymptomatic. Two patients had a recurrent vertebrobasilar TIA; there was no recurrent stroke. The initial DSA findings consisted of 14 stenoses, 20 tapered occlusions, and 6 pseudoaneurysms. During follow‐up, 63% of the vessels recanalized. Sonographic findings were consistent with angiographic findings in 80% at the initial examination and in 86% during follow‐up. The main reason for discordant results was the failure of CDUS to detect pseudoaneurysms. No recurrence occurred in the vertebral arteries (VA), but 1 patient had an asymptomatic carotid artery dissection during follow‐up.

Conclusion.

Recurrent TIA or stroke after VAD appears to be extremely rare, independent of recanalization or persistent occlusion of the affected artery. CDUS and TCCDUS provide reliable follow‐up of VAD in all patients presenting with stenosis or occlusion, but do not allow for detection of pseudoaneurysms of the VA. © 2008 Wiley Periodicals, Inc. J Clin Ultrasound, 2008
Hybridization betweenCrocodylus acutusandCrocodylus moreletiiin the Yucatan Peninsula: I. evidence from mitochondrial DNA and morphology
Authors: Cedeño‐Vázquez José Rogelio , Rodriguez David , Calmé Sophie , Ross James Perran , Densmore Llewellyn D. , Thorbjarnarson John B.
Published: July 2008

The American crocodile (Crocodylus acutus) and the Morelet's crocodile (C. moreletii) are broadly sympatric in Belize and Mexico. The presence of morphologically anomalous individuals in the overlapping range area suggests possible hybridization between these species. Analysis of 477 base pairs of the mitochondrial tRNAPro‐tRNAPhe‐Dloop region revealed the presence of pureC.acutus(N=43) andC. moreletii(N=56), as well as a high proportion of interspecific hybrids (N=17, 14.6%) in the Yucatan Peninsula, Mexico. Although all individuals could be assigned to one species or other based on phenotypic characters, some had been characterized as potential hybrids in the field by anomalous scale counts. The hybridization zone lies along the area of sympatry betweenC. acutusandC. moreletiiinvestigated in this study, but extends further inland if hybrid localities from Belize are included. Hybridization in the Yucatan Peninsula is bidirectional, which indicates considerably more genetic contact between these species than previously recognized, and is probably more detrimental to the genetic integrity of smallerC. acutuspopulations. A more intensive study of the pattern of hybridization is warranted and supports continued classification ofC. acutusas a critically threatened species in the Yucatan Peninsula.J. Exp. Zool. 309A:661–673, 2008. © 2008 Wiley‐Liss, Inc.
Spider silk aging: initial improvement in a high performance material followed by slow degradation
Authors: Agnarsson Ingi , Boutry Cecilia , Blackledge Todd A.
Published: July 2008

Spider silk possesses a unique combination of high tensile strength and elasticity resulting in extraordinarily tough fibers, compared with the best synthetic materials. However, the potential application of spider silk and biomimetic fibers depends upon retention of their high performance under a variety of conditions. Here, we report on changes in the mechanical properties of dragline and capture silk fibers from several spider species over periods up to 4 years of benign aging. We find an improvement in mechanical performance of silk fibers during the first year of aging. Fibers rapidly decrease in diameter, suggesting an increase in structural alignment and organization of molecules. One‐year old silk also is stiffer and has higher stress at yield than fresh silk, whereas breaking force, elasticity, and toughness either improve or are unaffected by early aging. However, 4‐year old silk shows signs of degradation as the breaking load, elasticity, and toughness are all lower than in fresh silk. Aging, however, does not reduce the tensile strength of silk. These data suggest initially rapid reorganization and tighter packaging of molecules within the fiber, followed by longer‐term decomposition. We hypothesize that possibly the breakdown of amino acids via emission of ammonia gas, as is seen in long‐term aging of museum silkworm fabrics, may contribute. Degradation of spider silk under benign conditions may be a concern for efforts to construct and utilize biomimetic silk analogs. However, our findings suggest an initial improvement in mechanical performance and that even old spider silk still retains impressive mechanical performance. J. Exp. Zool. 309A:494–504, 2008. © 2008 Wiley‐Liss, Inc.
Ectopic dermal ridge configurations on the interdigital webbings and postaxial marginal portion of the hindlimb in Hammertoe mutant mice (Hm)
Authors: Kimura Sumiko , Schaumann Blanka A. , Shiota Kohei
Published: July 2008

The effects of the hereditary malformation of Hammertoe mutant mice (gene symbol Hm) on the digital pads and dermal ridge configurations on their hindlimbs were examined. In the wild‐type (+/+) mice with normally separated digits, dermal ridges developed only on the pads. Heterozygous (Hm/+) and homozygous (Hm/Hm) mutant mice, however, had a broad big toe, fused interdigital soft tissues, reduced claws, an extra rudimentary postaxial digit and camptodactyly. The dermal ridges appeared not only on the pads, affected in their number and configurations, but also on the ventral surface of the interdigital webbings and postaxial marginal area exhibiting an extra rudimentary digit and webbing. These aberrant configurations may be related to the abnormal occurrence of programmed cell death (PCD) in the interdigital zones and the postaxial marginal portion in Hm/+ and Hm/Hm mice. That is, the diminished cell death may fail to decrease the cell density in the interdigital zones and postaxial marginal portion and result in the webbing and an extra rudimentary digit and webbing, respectively. Simultaneously, it could also interrupt the migration of surviving cells of these areas toward the neighboring digits and the distal area of the sole and produce the ectopic dermal ridges on the way to the as yet unformed (presumptive) digital and plantar volar pads. The present findings suggest that normal interdigital and pre/postaxial PCD contributes not only to the separation of digits, the initial formation of individual digits of different sizes, and the inhibition of the extra digit but also to the development of the presumptive digital and plantar pads, including dermal ridges. J. Morphol., 2008. © 2008 Wiley‐Liss, Inc.
Recombinant arginine deiminase reduces inducible nitric oxide synthase iNOS‐mediated neurotoxicity in a coculture of neurons and microglia
Authors: Yu Hao‐Hsin , Wu Fe‐Lin Lin , Lin Shan‐Erh , Shen Li‐Jiuan
Published: July 2008

Modulation of nitric oxide (NO) production is considered a promising approach to therapy of diseases involving excessive inducible nitric oxide synthase (iNOS) expression, such as certain neuronal diseases. Recombinant arginine deiminase (rADI, EC3.5.3.6) catalyzes the conversion of L‐arginine (L‐arg), the sole substrate of NOS for NO production, to L‐citrulline (L‐cit) and ammonia. To understand the effect of the depletion of L‐arg by rADI on NO concentration and neuroprotection, a direct coculture of neuron SHSY5Y cells and microglia BV2 cells treated with lipopolysaccharide (LPS) and interferon‐γ (IFN‐γ) was used as a model of iNOS induction. The results showed that rADI preserved cell viability (4‐fold higher compared with the cells treated with LPS/IFN‐γ only) by the MTT assay, corresponding with the results of neuronal viability by neuron‐specific immunostaining assay. NO production (mean ± SD) decreased from 67.0 ± 1.3 to 19.5 ± 5.5 μM after a 2‐day treatment of rADI by the Griess assay; meanwhile, induction of iNOS protein expression by rADI was observed. In addition, rADI substantially preserved the neuronal function of dopamine uptake in the coculture. The replenishment of L‐arg in the coculture eliminated the neuroprotective and NO‐suppressive effects of rADI in the coculture, indicating that L‐arg played a crucial role in the effects of rADI. These results highlight the important role of L‐arg in the neuron‐microglia coculture in excessive induction of iNOS. Regulation of L‐arg by ADI demonstrated that rADI has a potentially therapeutic role in iNOS‐related neuronal diseases. © 2008 Wiley‐Liss, Inc.
Light and food signals cooperate to entrain the rat pineal circadian system
Authors: Wu Tao , Jin Yuanxiang , Kato Hisanori , Fu Zhengwei
Published: July 2008

Light and food signals are two dominant Zeitgeber for central and peripheral circadian clocks, respectively. However, whether and how changes in light and food stimuli affect circadian rhythms of mammalian pineal clock genes are still largely unknown. For the first time, in the present study, we established a method for analysis of the expression levels of most clock genes (Bmal1, Per1, Per2, Per3, Cry1, Dec1 and Dec2) and AANAT in a single rat pineal gland and used it to investigate individual and cooperative effects of feeding and light cycle reversals on the pineal circadian system. Feeding reversal (RF) alone for 7 days had only weak effect on the rhythmicity of clock gene expression in the pineal gland, whereas light/dark reversal for the same period without any change in the feeding regime could not absolutely reverse the expression phases of all examined genes either. However, reversal of the LD cycle together with the feeding regime completely resynchronized the circadian gene expression in the pineal gland within 7 days. These data suggest that photic regulation is the dominant cue that synchronizes the pineal circadian rhythm more efficiently than the feeding‐related cue, but food availability does contribute to reentrainment of the pineal circadian system. In addition, the inability of RF in the pineal circadian system suggests that the pineal gland is more strongly coupled to the master clock than other peripheral organs and alternatively distinguishes it from other peripheral organs in the hierarchically organized mammalian circadian systems. © 2008 Wiley‐Liss, Inc.
Fibroblast growth factor‐2 mRNA expression in the brainstem and spinal cord of normal and chronic spinally transected urodeles
Authors: Moftah Marie , Landry Marc , Nagy Frédéric , Cabelguen Jean‐Marie
Published: July 2008

Descending pathways in the spinal cord of adult urodele amphibians show a high regenerative ability after body spinal cord transection; regenerated axons regrow into the transected spinal cord, and hindlimb locomotor recovery occurs spontaneously. Little is currently known about the molecular basis of spinal cord regeneration in urodeles, but it is believed that fibroblast growth factor‐2 (FGF2) may play an important role by inducing proliferation of neural progenitor cells. The aim of our study, using in situ hybridization in adult Pleurodeles waltlii, was twofold: 1) to document FGF2 mRNA expression pattern along the brainstem‐spinal cord of intact salamanders and 2) to investigate the changes in this pattern in animals unable to display hindlimb locomotor movements and in animals having fully recovered hindlimb locomotor activity after body spinal cord transection. This design establishes a firm basis for further studies on the role of FGF2 in functional recovery of hindlimb locomotion. Our results revealed a decreasing rostrocaudal gradient in FGF2 mRNA expression along the brainstem‐spinal cord in intact animals. They further demonstrated a long‐lasting up‐regulation of FGF2 mRNA expression in response to spinal transection at the midtrunk level, both in brainstem and in the spinal cord below the injury. Finally, double immunolabeling showed that FGF2 was up‐regulated in neuroglial, presumably undifferentiated, cells. Therefore, we propose that FGF2 may be involved in cell proliferation and/or neuronal differentiation after body spinal cord transection in salamander and could thus play an important role in functional recovery of locomotion after spinal lesion. © 2008 Wiley‐Liss, Inc.
Topology and patch‐clamp analysis of the sodium channel in relationship to the anti‐lipid a antibody in campylobacteriosis
Authors: Usuki Seigo , Nakatani Yoshihiko , Taguchi Kyoji , Fujita Tetsuhiro , Tanabe Shinya , Ustunomiya Iku , Gu Yihua , Cawthraw Shaun A. , Newell Diane G. , Pajaniappan Mohanasundari , Thompson Stuart A. , Ariga Toshio , Yu Robert K.
Published: July 2008

An infecting strain VLA2/18 of Campylobacter jejuni was obtained from an individual with campylobacteriosis and used to prepare chicken sera by experimental infection to investigate the role of serum anti‐ganglioside antibodies in Guillain‐Barré syndrome. Both sera of the patient and chicken contained anti‐ganglioside antibodies and anti‐Lipid A (anti‐Kdo2‐Lipid A) antibodies directed against the lipid A portion of the bacterial lipooligosaccharide. The anti‐Kdo2‐Lipid A activities inhibited voltage‐gated Na (Nav) channel of NSC‐34 cells in culture. We hypothesized that anti‐Kdo2‐Lipid A antibody acts on the functional inhibition of Nav1.4. To test this possibility, a rabbit peptide antibody (anti‐Nav1.4 pAb) against a 19‐mer peptide (KELKDNHILNHVGLTDGPR) on the α subunit of Nav1.4 was produced. Anti‐Nav1.4 pAb was cross‐reactive to Kdo2‐Lipid A. Anti‐Kdo2‐lipid A antibody activity in the chicken serum was tested for the Na+ current inhibition in NSC‐34 cells in combination with μ‐Conotoxin and tetrodotoxin. Contrary to our expectations, the anti‐Kdo2‐Lipid A antibody activity was extended to Nav channels other than Nav1.4. By overlapping structural analysis, it was found that there might be multiple peptide epitopes containing certain dipeptides showing a structural similarity with v‐Lipid A. Thus, our study suggests the possibility that there are multiple epitopic peptides on the extracellular domains of Nav1.1 to 1.9, and some of them may represent target sites for anti‐Kdo2‐Lipid A antibody, to induce neurophysiological changes in GBS by disrupting the normal function of the Nav channels. © 2008 Wiley‐Liss, Inc.
Cloning, expression, and functional analysis of rhesus monkey trace amine‐associated receptor 6: Evidence for lack of monoaminergic association
Authors: Xie Zhihua , Vallender Eric J. , Yu Naichen , Kirstein Shelli L. , Yang Hong , Bahn Mary E. , Westmoreland Susan V. , Miller Gregory M.
Published: July 2008

Several recent studies report an association between trace amine‐associated receptor 6 (TAAR6) and susceptibility to schizophrenia and bipolar affective disorder in humans. However, endogenous TAAR6 agonists and the receptor signaling profile and brain distribution remain unclear. Here, we clone TAAR6 from the rhesus monkey and use transfected cells to investigate whether this receptor interacts with brain monoamines and a psychostimulant drug to trigger cAMP signaling or extracellular signal‐regulated kinase (ERK) phosphorylation, while investigating its expression profile in the rhesus monkey brain. Unlike TAAR1, rhesus monkey TAAR6 did not alter cAMP levels in response to 10 μM of monoamines (dopamine, norepinephrine, serotonin, β‐phenylethylamine (β‐PEA), octopamine, tryptamine, and tyramine) or methamphetamine in stably transfected cells in vitro. Real‐time cell electronic sensing analysis indicated that the receptor did not alter cell impedance or change the effect of forskolin on cell impedance at exposure to 20 μM of each monoamine, suggesting a lack of either Gs or Gi‐linked signaling. Whereas κ opioid receptor activation led to ERK phosphorylation at exposure to 1 μM U69593, rhesus monkey TAAR6 had no such effect at exposure to 10 μM of monoamines or methamphetamine. Membrane and cell surface localization of TAAR6 was confirmed by immunocytochemistry, biotinylation, and Western blot testing with a TAAR6 antibody in the transfected cells. Real‐time reverse transcriptase–polymerase chain reaction amplification showed that TAAR6 mRNA was undetectable in selected rhesus monkey brain regions. Together, the data reveal that TAAR6 is unresponsive to brain monoamines and is not expressed in rhesus monkey brain monoaminergic nuclei, suggesting TAAR6 lacks direct association with brain monoaminergic neuronal function. © 2008 Wiley‐Liss, Inc.
Histamine‐induced Ca2+ entry in human astrocytoma U373 MG cells: Evidence for involvement of store‐operated channels
Authors: Barajas Margarita , Andrade Arturo , Hernandez‐Hernandez Oscar , Felix Ricardo , Arias‐Montaño Jose‐Antonio
Published: July 2008

Glial and glia‐derived cells express a variety of receptors for neurotransmitters and hormones, the majority of which evoke both Ca2+ release from intracellular stores and Ca2+ entry across the plasma membrane. We investigated the links between histamine H1 receptor activation, Ca2+ release from intracellular stores and Ca2+ influx in human astrocytoma U373 MG cells. Histamine, through a H1 receptor‐mediated effect, evoked an increase in cytoplasmic free calcium concentration ([Ca2+]i) that occurred in two phases: an initial, transient, increase owing to Ca2+ mobilization from intracellular pools, and a second, sustained increase dependent on both Ca2+ influx and continuous receptor occupancy. The characteristics of histamine‐induced increases in [Ca2+]i were similar to the capacitative entry evoked by emptying of the Ca2+ stores with thapsigargine, and different from that observed when Ca2+ influx was activated with OAG (1‐oleoyl‐2‐acetyl‐sn‐glycerol), a diacylglycerol (DAG) analog. OAG application or increased endogenous DAG, resulting from DAG kinase inhibition, reduced the histamine‐induced response. Furthermore, activation of the DAG target, protein kinase C (PKC), by TPA (12‐O‐tetradecanoyl 4β‐phorbol 13α‐acetate) resulted in inhibition of the histamine‐induced Ca2+ response, an action prevented by PKC inhibitors. By using reverse transcriptase–polymerase chain reaction analysis, mRNAs for transient receptor potential channels (TRPCs) 1, 4, and 6 as well as for STIM1 (stromal‐interacting molecule) and Orai1 were found to be expressed in the U373 MG cells, and confocal microscopy using specific antibodies revealed the presence of the corresponding proteins. Therefore, TRPCs may be candidate proteins forming store‐operated channels in the U373 MG cell line. Further, our results confirm the involvement of PKC in the regulation of H1 receptor‐induced responses and point out to the existence of a feedback mechanism acting via PKC to limit the increase in [Ca2+]i. © 2008 Wiley‐Liss, Inc.
Acute brain cytokine responses after global birth hypoxia in the rat
Authors: Ashdown Helen , Joita Silvia , Luheshi Giamal N. , Boksa Patricia
Published: July 2008

The main cause of hypoxic/ischemic brain damage in term human neonates is intrauterine asphyxia, in which the whole body is subjected to hypoxia. Inflammatory cytokines are thought to play an important role in modulating hypoxic/ischemic damage in immature brain. Evidence for this from animal models is based mainly on studies that used a model of carotid artery ligation with hypoxia in postnatal rats. However, little is known about the role of cytokines in brain injury after whole‐body hypoxia at the time of birth. This study used a well‐established rat model of global birth hypoxia to assess mRNA and protein expression of three key proinflammatory cytokines, interleukin‐1β (IL‐1β), IL‐6, and tumor necrosis factor‐α, (TNF‐α) in the brain, liver, and kidney of neonates. We observed decreased IL‐1β and TNF‐α protein, and decreased IL‐6 mRNA in brains of neonates in the 2 hr after birth hypoxia but increased IL‐6 and IL‐1β in liver compared with vaginally born controls. Increasing the severity of the insult by increasing the period of anoxic exposure further decreased brain IL‐1β, whereas delivering anoxia under hypothermic conditions, known to be neuroprotective, attenuated the decrease in brain IL‐1β. These data suggest that decreased brain levels of inflammatory cytokines may modulate central nervous system responses to global birth hypoxia in rats. Our findings of decreased brain cytokine expression after global birth hypoxia contrast with reports of increased brain cytokines after carotid artery ligation with hypoxia in postnatal rats; possible reasons for these differences are discussed. © 2008 Wiley‐Liss, Inc.
Defective insulin signaling pathway and increased glycogen synthase kinase‐3 activity in the brain of diabetic mice: Parallels with Alzheimer's disease and correction by insulin
Authors: Jolivalt C.G. , Lee C.A. , Beiswenger K.K. , Smith J.L. , Orlov M. , Torrance M.A. , Masliah E.
Published: July 2008

We have evaluated the effect of peripheral insulin deficiency on brain insulin pathway activity in a mouse model of type 1 diabetes, the parallels with Alzheimer's disease (AD), and the effect of treatment with insulin. Nine weeks of insulin‐deficient diabetes significantly impaired the learning capacity of mice, significantly reduced insulin‐degrading enzyme protein expression, and significantly reduced phosphorylation of the insulin‐receptor and AKT. Phosphorylation of glycogen synthase kinase‐3 (GSK3) was also significantly decreased, indicating increased GSK3 activity. This evidence of reduced insulin signaling was associated with a concomitant increase in tau phosphorylation and amyloid β protein levels. Changes in phosphorylation levels of insulin receptor, GSK3, and tau were not observed in the brain of db/db mice, a model of type 2 diabetes, after a similar duration (8 weeks) of diabetes. Treatment with insulin from onset of diabetes partially restored the phosphorylation of insulin receptor and of GSK3, partially reduced the level of phosphorylated tau in the brain, and partially improved learning ability in insulin‐deficient diabetic mice. Our data indicate that mice with systemic insulin deficiency display evidence of reduced insulin signaling pathway activity in the brain that is associated with biochemical and behavioral features of AD and that it can be corrected by insulin treatment. © 2008 Wiley‐Liss, Inc.
Quality of fresh‐cut tomatoes as affected by salt content in irrigation water and post‐processing ultraviolet‐C treatment
Authors: Kim Hyun‐Jin , Fonseca Jorge M , Kubota Chieri , Kroggel Mark , Choi Ju‐Hee
Published: July 2008

BACKGROUND: There is little information available concerning the effect of sanitizing agents, including ultraviolet (UV‐C) light, on nutritional composition of fresh‐cut produce. In this study, the influence of UV‐C light irradiation (3.2–19.2 kJ m−2) on the nutritional quality of fresh‐cut tomato cv. Durinta grown under either low (2.4/2.8 dS m−1) or high (4.9/7.7 dS m−1) electrical conductivity (EC) of the hydroponic nutrient influx/efflux solution was investigated.

RESULTS: When grown under low EC UV‐C light treatment reduced development of microbial population of fresh‐cut tomato, increased phenolic content and delayed degradation of vitamin C after 7 days of storage at 4–6 °C. UV‐C light irradiation had no significant effect on appearance, color or lycopene content of fresh‐cut tomato. In recently harvested intact tomatoes, the lycopene and vitamin C contents were 30% higher in tomatoes irrigated with high EC, whereas the phenolic content was unaffected by the salt treatment. After fresh‐cut processing, phenolic and vitamin C contents of fresh cuts produced from tomatoes treated with high EC decreased by > 10%. This decline was initially accelerated in tomato treated with UV‐C light irradiation.

CONCLUSIONS: Our results revealed that fresh‐cut tomatoes are affected dissimilarly by UV‐C treatments, depending on the degree of salt stress undergone by plants before harvest. We determined the impact of a sanitizing procedure on nutritional composition, an effect often overlooked but with great significance to ensure overall quality of fresh produce. Our findings revealed the need for more nutritional studies addressing the effect of multiple stress factors from pre‐harvest to post‐processing stages. Copyright © 2008 Society of Chemical Industry
Chlorophyll degradation and resulting catabolite formation in stored Japanese bunching onion (Allium fistulosum L.)
Authors: Dissanayake Prasajith K , Yamauchi Naoki , Shigyo Masayoshi
Published: July 2008

BACKGROUND: Loss of green colour is the main factor in quality deterioration of stored Japanese bunching onion (JBO, Allium fistulosum L.) leaves. Elucidation of chlorophyll (Chl) degradation has not been performed to date. In the present study, Chl catabolism and Chl‐degrading enzyme activities in stored JBO leaves were investigated.

RESULTS: Green leaves of JBO stored at 25 °C turned yellow within 3 days, whereas no changes occurred at 4 °C. Pheophytin (Phy) a, chlorophyllide (Chlide) a, pheophorbide (Pheide) a and C132‐hydroxychlorophyll (OHChl) a were the main catabolites of Chl a and diminished at 25 °C concomitant with leaf yellowing, whereas no significant reductions were observed at 4 °C. The activities of Chl‐degrading peroxidase and Mg‐dechelation increased significantly at 25 °C but not at 4 °C.

CONCLUSION: Chl degradation was accelerated and catabolite levels (mainly Phy a, Chlide a, Pheide a and OHChl a) were diminished in JBO leaves stored at 25 °C, suggesting that the catabolites are rapidly converted into their following forms. Activities of Mg‐dechelation and Chl‐degrading peroxidase were significantly enhanced during storage at 25 °C, indicating that these two enzymes have major roles in Chl degradation in stored JBO leaves. Copyright © 2008 Society of Chemical Industry
In vitro gas production profiles and fermentation end‐products in processed peas, lupins and faba beans
Authors: Azarfar Arash , Tamminga Seerp , Pellikaan WF , van der Poel Antonius FB
Published: July 2008

BACKGROUND: An experiment was carried out to establish whether using a pre‐compacting device (expander) changes the contribution of dry matter (DM) and degradative behaviour of peas, lupins and faba beans over the different fractions (non‐washable fraction, NWF; insoluble washable fraction, ISWF; soluble washable fraction, SWF). Samples of the entire concentrate ingredients (WHO ingredients) and their different fractions (NWF, ISWF and SWF) were subjected to three processes (Retsch milling, R; expander treatment, E; expander‐pelleting, EP) and their fermentation characteristics were evaluated using an in vitro gas production technique.

RESULTS: In peas and faba beans, both the E and EP processes increased the size of the NWF (P < 0.05) and decreased the size of the SWF compared with the R process. The maximum fractional rate of gas production in the first phase of fermentation was higher in the E and EP samples than in the R samples (P < 0.05). In lupins and faba beans the E and EP processes shifted the pattern of fermentation towards a more glucogenic fermentation, as represented by a lower non‐glucogenic to glucogenic ratio (NGR). Ammonia production (NH3‐N) in the E and EP samples was significantly (P < 0.05) lower than that in the R samples.

CONCLUSION: It is concluded that the E and EP processes provide a certain level of protection against ruminal breakdown to dietary protein and shift the pattern of fermentation towards a more glucogenic fermentation. Copyright © 2008 Society of Chemical Industry
Composition and radical‐scavenging activity of Thymus glabrescens Willd. (Lamiaceae) essential oil
Authors: Maksimović Zoran , Stojanović Danilo , Šoštarić Ivan , Dajić Zora , Ristić Mihailo
Published: July 2008

BACKGROUND: Knowledge about the chemical and therapeutic properties of Thymus glabrescens Willd. (Lamiaceae) is scarce and inconsistent. Therefore the main objectives of this study were to determine the yield and chemical composition of essential oils from wild‐growing T. glabrescens populations, to assess their radical scavenging activity and to correlate the results with published data in order to deduce which components are responsible for the activity.

RESULTS: The plant material yielded between 4.0 and 8.0 mL kg−1 of essential oil. All samples contained considerable but variable concentrations of thymol (22.3–55.1%), depending on the source. Radical‐scavenging activities of the oils were estimated by 1,1‐diphenyl‐2‐picrylhydrazyl free radical (DPPH) assay against butylated hydroxytoluene (BHT) and thymol as positive controls. The observed activities (IC50 values ranged from 94 to 230 µg mL−1) were strongly influenced by thymol concentration, as verified by rapid screening for DPPH radical‐scavenging activity on thin layer chromatography (TLC) plates and regression analysis.

CONCLUSION: These results represent the first report on the free radical‐scavenging activity of T. glabrescens essential oil and one of the first comprehensive reports on its composition. Thymus glabrescens could be used in the food industry for seasoning purposes or for preserving processed foods from oxidative degradation. Copyright © 2008 Society of Chemical Industry
Fertilization in vitro with spermatozoa from different mice increased variation in the developmental potential of embryos compared to artificial parthenogenetic activation
Authors: Miao De‐Qiang , Liang Bo , Wang Jun‐Zuo , Wang Hui‐Li , Cui Wei , Liu Yong , Tan Jing‐He
Published: July 2008

Although successful embryo development is dependent upon genetic and epigenetic contributions from both the male and female, the male potential to adversely affect embryo development has been scarcely studied. It is unclear whether the sperm variation among different males would affect the outcome of oocyte evaluation by embryo development following fertilization. In the present study, variation in the developmental potential of mouse embryos was first compared between in vitro fertilization with epididymal spermatozoa from different males and Sr2+ parthenogenetic activation using oocytes of different qualities, and then the effect of male on fertilization and embryo development was examined using randomly chosen oocytes and spermatozoa from cauda epididymidis, vas deferens or electro‐ejaculates. Rates of fertilization and blastocyst formation were significantly higher with spermatozoa from cauda epididymidis or vas deferens than with ejaculated spermatozoa. Rates of embryonic development differed significantly between different males, but not between different ejaculates of the same male. Analysis of standard errors of means and coefficients of variance indicated that as long as multiple males were involved, the variation in oocyte fertilization/activation and blastocyst formation was always higher after fertilization than after Sr2+ parthenogenetic activation whether spermatozoa were collected from epididymidis, vas deferens or ejaculates and regardless of oocyte qualities. It is concluded that (1) epididymal mouse spermatozoa fertilize more oocytes than ejaculated spermatozoa under identical experimental conditions; (2) like farm animals, the mice also show a remarkable male effect on the developmental potential of in vitro produced embryos although they are supposed to be less genetically diverse; (3) parthenogenetic activation is recommended for assessment of oocyte quality to exclude the effect of male. Mol. Reprod. Dev. 76: 239–245, 2009. © 2008 Wiley‐Liss, Inc.
The role of annexins in tumour development and progression
Authors: Mussunoor S , Murray GI
Published: July 2008

The annexins are a super‐family of closely related calcium and membrane‐binding proteins. They have a diverse range of cellular functions that include vesicle trafficking, cell division, apoptosis, calcium signalling and growth regulation. Many studies have shown the annexins to be among the genes whose expression are consistently differentially altered in neoplasia. Some annexins show increased expression in specific types of tumours, while others show loss of expression. Mechanistic studies relating the changes in annexin expression to tumour cell function, particularly tumour invasion and metastasis, angiogenesis and drug resistance, are now also emerging. Changes in the expression of individual annexins are associated with particular types of tumour and hence the annexins may also be useful biomarkers in the clinic. Copyright © 2008 Pathological Society of Great Britain and Ireland. Published by John Wiley & Sons, Ltd.
Suppression of putative tumour suppressor gene GLTSCR2 expression in human glioblastomas
Authors: Kim Y‐J , Cho Y‐E , Kim Y‐W , Kim J‐Y , Lee S , Park J‐H
Published: July 2008

Glioma tumour‐suppressor candidate region gene 2 (GLTSCR2/PICT‐1) is localized within the well‐known 1.4 Mb tumour‐suppressive region of chromosome 19q, which is frequently altered in various human tumours, including diffuse gliomas. Aside from its chromosomal localization, several lines of evidence, including PTEN‐phosphorylating and cell‐killing activities, suggests that GLTSCR2 participates in the suppression of tumour growth and development. However, little is known about the biological functions and molecular mechanisms of GLTSCR2 as a tumour suppressor gene. We investigated the pathological significance of GLTSCR2 expression in association with the development and progression of glioblastomas, the most common malignant brain tumour. We used real‐time PCR and western blot analysis to examine the expression levels of GLTSCR2 mRNA and protein in glioblastomas, normal brain tissue and in non‐glial tumour tissue of different origin, and found that GLTSCR2 expression is down‐regulated in glioblastomas. In addition, direct sequencing analysis and fluorescence in situ hybridization clearly demonstrates the presence of genetic alterations, such as a nonsense mutation and deletion, in the GLTSCR2 gene in glioblastomas. Finally, our immunohistochemical study demonstrates that GLTSCR2 is sequentially down‐regulated according to the histological malignant progression of the astrocytic glial tumour. Taken together, our results suggest that GLTSCR2 is involved in astrocytic glioma progression. Copyright © 2008 Pathological Society of Great Britain and Ireland. Published by John Wiley & Sons, Ltd.
Increased KIT signalling with up‐regulation of cyclin D correlates to accelerated proliferation and shorter disease‐free survival in gastrointestinal stromal tumours (GISTs) with KIT exon 11 deletions
Authors: Haller F , Löbke C , Ruschhaupt M , Schulten H‐J , Schwager S , Gunawan B , Armbrust T , Langer C , Ramadori G , Sültmann H , Poustka A , Korf U , Füzesi L
Published: July 2008

Gastrointestinal stromal tumours (GISTs) with deletions in KIT exon 11 are characterized by higher proliferation rates and shorter disease‐free survival times, compared to GISTs with KIT exon 11 point mutations. Up‐regulation of cyclin D is a crucial event for entry into the G1 phase of the cell cycle, and links mitogenic signalling to cell proliferation. Signalling from activated KIT to cyclin D is directed through the RAS/RAF/ERK, PI3K/AKT/mTOR/EIF4E, and JAK/STATs cascades. ERK and STATs initiate mRNA transcription of cyclin D, whereas EIF4E activation leads to increased translation efficiency and reduced degradation of cyclin D protein. The aim of the current study was to analyse the mRNA and protein expression as well as protein phosphorylation of central hubs of these signalling cascades in primary GISTs, to evaluate whether tumours with KIT exon 11 deletions and point mutations differently utilize these pathways. GISTs with KIT exon 11 deletions had significantly higher mitotic counts, higher proliferation rates, and shorter disease‐free survival times. In line with this, they had significantly higher expression of cyclin D on the mRNA and protein level. Furthermore, there was a significantly higher amount of phosphorylated ERK1/2, and a higher protein amount of STAT3, mTOR, and EIF4E. PI3K and phosphorylated AKT were also up‐regulated, but this was not significant. Ultimately, GISTs with KIT exon 11 deletions had significantly higher phosphorylation of the central negative cell‐cycle regulator RB. Phosphorylation of RB is accomplished by activated cyclin D/CDK4/6 complex, and marks a central event in the release of the cell cycle. Altogether, these observations suggest increased KIT signalling with up‐regulation of cyclin D as the basis for the unfavourable clinical course in GISTs with KIT exon 11 deletions. Copyright © 2008 Pathological Society of Great Britain and Ireland. Published by John Wiley & Sons, Ltd.