bluuebunny/crossref_metadata_2025 huggingface dataset:
3,4‐Methylenedioxymethamphetamine increases interleukin‐1β levels and activates microglia in rat brain: studies on the relationship with acute hyperthermia and 5‐HT depletion
Authors: Orio Laura , O'Shea Esther , Sanchez Veronica , Pradillo Jesus M. , Escobedo Isabel , Camarero Jorge , Moro Maria A. , Green A. Richard , Colado M. Isabel
Published: June 2004

3,4‐Methylenedioxymethamphetamine (MDMA) administration to rats produces acute hyperthermia and 5‐HT release. Interleukin‐1β (IL‐1β) is a pro‐inflammatory pyrogen produced by activated microglia in the brain. We examined the effect of a neurotoxic dose of MDMA on IL‐1β concentration and glial activation and their relationship with acute hyperthermia and 5‐HT depletion. MDMA, given to rats housed at 22°C, increased IL‐1β levels in hypothalamus and cortex from 1 to 6 h and [3H]‐(1‐(2‐chlorophenyl)‐N‐methyl‐N‐(1‐methylpropyl)3‐isoquinolinecarboxamide) binding between 3 and 48 h. Increased immunoreactivity to OX‐42 was also detected. Rats became hyperthermic immediately after MDMA and up to at least 12 h later. The IL‐1 receptor antagonist did not modify MDMA‐induced hyperthermia indicating that IL‐1β release is a consequence, not the cause, of the rise in body temperature. When MDMA was given to rats housed at 4°C, hyperthermia was abolished and the IL‐1β increase significantly reduced. The MDMA‐induced acute 5‐HT depletion was prevented by fluoxetine coadministration but the IL‐1β increase and hyperthermia were unaffected. Therefore, the rise in IL‐1β is not related to the acute 5‐HT release but is linked to the hyperthermia. Contrary to IL‐1β levels, microglial activation is not significantly modified when hyperthermia is prevented, suggesting that it might be a process not dependent on the hyperthermic response induced by MDMA.
Increased expression of the gene for the Y1 receptor of neuropeptide Y in the amygdala and paraventricular nucleus of Y1R/LacZ transgenic mice in response to restraint stress
Authors: Mele Paolo , Oberto Alessandra , Serra Mariangela , Pisu Maria Giuseppina , Floris Ivan , Biggio Giovanni , Eva Carola
Published: June 2004

A sustained increase in the brain concentrations of neuroactive steroids was previously shown to induce Y1 receptor gene expression in the amygdala of Y1R/LacZ transgenic mice which harbour a construct comprising the murine Y1 receptor gene promoter and the lacZ reporter gene. We now investigated the effects of restraint stress on both the cerebrocortical concentrations of neuroactive steroids and Y1 receptor gene expression in the amygdala and hypothalamic paraventricular nucleus (PVN) of Y1R/LacZ transgenic mice. The cerebrocortical concentrations of allopregnanolone and allotetrahydrodeoxycorticosterone were significantly increased immediately after a 1‐h exposure to restraint stress and had returned to control values within 30 min. Expression of Y1R/LacZ was increased in the amygdala and PVN 6 h after restraint. The 5α‐reductase inhibitor finasteride, that prevented the increase in neuroactive steroid concentrations, did not block that in transgene expression induced by 1‐h restraint. Daily exposure to restraint for 10 days also increased the cerebrocortical concentrations of neuroactive steroids but failed to affect transgene expression. Acute but not repeated restraint thus increases Y1 receptor gene expression in the amygdala and PVN, suggesting that tolerance develops towards this stressor. The effect of acute restraint is not mediated by the increase in the brain concentrations of neuroactive steroids but may rather reflect a ligand‐induced increase in Y1 receptor gene transcription. Data support a role of Y1 receptors in the behavioural and neuroendocrine responses to stress.
Parkin attenuates manganese‐induced dopaminergic cell death
Authors: Higashi Youichirou , Asanuma Masato , Miyazaki Ikuko , Hattori Nobutaka , Mizuno Yoshikuni , Ogawa Norio
Published: June 2004

Manganese as environmental factor is considered to cause parkinsonism and induce endoplasmic reticulum stress‐mediated dopaminergic cell death. We examined the effects of manganese on parkin, identified as the gene responsible for familial Parkinson's disease, and the role of parkin in manganese‐induced neuronal cell death. Manganese dose‐dependently induced cell death of dopaminergic SH‐SY5Y and CATH.a cells and cholinergic Neuro‐2a cells, and that the former two cell types were more sensitive to manganese toxicity than Neuro‐2a cells. Moreover, manganese increased the expression of endoplasmic reticulum stress‐associated genes, including parkin, in SH‐SY5Y cells and CATH.a cells, but not in Neuro‐2a cells. Treatment with manganese resulted in accumulation of parkin protein in SH‐SY5Y cells and its redistribution to the perinuclear region, especially aggregated Golgi complex, while in Neuro‐2a cells neither expression nor redistribution of parkin was noted. Manganese showed no changes in proteasome activities in either cell. Transient transfection of parkin gene inhibited manganese‐ or manganese plus dopamine‐induced cell death of SH‐SY5Y cells, but not of Neuro‐2a cells. Our results suggest that the attenuating effects of parkin against manganese‐ or manganese plus dopamine‐induced cell death are dopaminergic cell‐specific compensatory reactions associated with its accumulation and redistribution to perinuclear regions but not with proteasome system.
Dopamine D2 receptor activates extracellular signal‐regulated kinase through the specific region in the third cytoplasmic loop
Authors: Takeuchi Yusuke , Fukunaga Kohji
Published: June 2004

To investigate whether the third cytoplasmic loop and the C‐terminal cytoplasmic tail of dopamine D2 receptor (D2R) are involved in extracellular signal‐regulated kinase (ERK) activation and subsequent regulation of transcription factors, we established NG108‐15 cells stably expressing D2LR and D2SR deleted 40 amino acid residues in the third cytoplasmic loop (NGD2LR‐3rd‐dele and NGD2SR‐3rd‐dele) or the C‐terminal cytoplasmic tail (NGD2LR‐C‐dele and NGD2SR‐C‐dele) and evaluated these receptors' functions using luciferase reporter gene assay. Immunocytochemical studies showed similar intracellular distributions of D2LR‐3rd‐dele and D2SR‐3rd‐dele to D2LR and D2SR, respectively. Quinpirole‐induced inhibition of forskolin‐induced cyclic AMP responsive element (CRE) activation was not affected by the deletion of 40 amino acid residues. However, nuclear factor‐kappa B (NF‐κB) activation by D2R‐3rd‐dele was largely attenuated compared to that by D2R. Similarly, ERK or serum‐responsive element (SRE) activation by quinpirole treatment was totally abolished in NGD2R‐3rd‐dele cells. Moreover, D2R‐C‐dele was diffusely distributed or clustered in the cell bodies and lost the receptor functions. Taken together, the 40 amino acid residues in the third cytoplasmic loop are essential for the ERK activation but not for inhibition of adenylyl cyclase through Gi/o proteins. In addition, the C‐terminal cytoplasmic tail is essential for membrane association of D2Rs to elicit the receptor functions.
Immature nicastrin stabilizes APH‐1 independent of PEN‐2 and presenilin: identification of nicastrin mutants that selectively interact with APH‐1
Authors: Shirotani Keiro , Edbauer Dieter , Kostka Marcus , Steiner Harald , Haass Christian
Published: June 2004

γ‐Secretase is a high molecular mass aspartyl protease complex composed of presenilin (PS1 or PS2), nicastrin (Nct), anterior pharynx‐defective‐1 (APH‐1) and presenilin enhancer‐2 (PEN‐2). The complex mediates the intramembraneous proteolysis of β‐secretase cleaved β‐amyloid precursor protein (APP) leading to the secretion of the Alzheimer's disease‐associated amyloid β‐peptide (Aβ). In order to dissect functionally important domains of Nct required for γ‐secretase complex assembly, maturation, and activity we mutated evolutionary conserved amino acids. The mutant Nct variants were expressed in a cellular background with significantly reduced endogenous Nct. Mutant Nct was functionally investigated by its ability to restore PS, APH‐1 and PEN‐2 expression as well as by monitoring the accumulation of the APP C‐terminal fragments, the immediate substrates of γ‐secretase. We identified three independent mutations within the ectodomain of Nct, which rescued expression of APH‐1 but not of PEN‐2 or PS and thus failed to restore γ‐secretase activity. Interestingly, these immature Nct variants selectively bound to APH‐1, suggesting a stable Nct/APH−1 interaction independent of PS and PEN‐2. Consistent with this finding, expression of APH‐1 remained largely unaffected in the PS double knock‐out and immature Nct co‐immunoprecipitated with APH‐1 in the absence of PS and PEN‐2. Taken together, our findings suggest that immature Nct can stably interact with APH‐1 to form a potential scaffold for binding of PS and PEN‐2. Moreover, binding of the latter two complex partners critically depends on the integrity of the Nct ectodomain.
Neuroprotective effects of the mGlu5R antagonist MPEP towards quinolinic acid‐induced striatal toxicity: involvement of pre‐ and post‐synaptic mechanisms and lack of direct NMDA blocking activity
Authors: Popoli Patrizia , Pintor Annita , Tebano Maria Teresa , Frank Claudio , Pepponi Rita , Nazzicone Valeria , Grieco Rosa , Pèzzola Antonella , Reggio Rosaria , Minghetti Luisa , De Berardinis Maria Anna , Martire Alberto , Potenza Rosa Luisa , Domenici Maria Rosaria , Massotti Marino
Published: June 2004

The aim of this work was to investigate the potential neuroprotective effects of the metabotropic glutamate receptor 5 (mGlu5R) antagonist 2‐Methyl‐6‐(phenylethynyl)‐pyridine (MPEP) towards quinolinic acid (QA)–induced striatal excitoxicity. Intrastriatal MPEP (5 nmol/0.5 µL) significantly attenuated the body weight loss, the electroencephalographic alterations, the impairment in spatial memory and the striatal damage induced by bilateral striatal injection of QA (210 nmol/0.7 µL). In a second set of experiments, we aimed to elucidate the mechanisms underlying the neuroprotective effects of MPEP. In microdialysis studies in naive rats MPEP (80–250 µm through the dialysis probe) significantly reduced the increase in glutamate levels induced by 5 mm QA. In primary cultures of striatal neurons MPEP (50 µm) reduced the toxicity induced by direct application of glutamate [measured as release of lactate dehydrogenase [LDH]). Finally, we found that 50 µm MPEP was unable to directly block NMDA‐induced effects (namely field potential reduction in corticostriatal slices, as well as LDH release and intracellular calcium increase in striatal neurons). We conclude that: (i) MPEP has neuroprotective effects towards QA‐induced striatal excitotoxicity; (ii) both pre‐ and post‐synaptic mechanisms are involved; (iii) the neuroprotective effects of MPEP do not appear to involve a direct blockade of NMDA receptors.
A novel lipopolysaccharide‐modulated Jun binding repressor in intron 2 of CYP2E1
Authors: Tindberg Niclas , Bengtsson Inger , Hu Yin
Published: June 2004

Cytochrome P450 2E1 (CYP2E1) exhibits a pronounced oxidase activity that may mediate apoptotic injury in glial cells as well as hepatocytes. Strict regulation of CYP2E1 and it's activity is therefore thought to be crucial. We have studied CYP2E1 transcriptional regulation in primary cortical glial cells and have identified a novel repressor element at +1452/+1460 in intron 2 of the rat CYP2E1 gene. The element very potently repressed CYP2E1 and SV40 promoters and consisted of the non‐palindromic core sequence 5′‐TTCCACTCA‐3′. Jun proteins were found to interact with the site. The protein complexes were also found to contain an as yet unidentified protein of ≈60 kDa, probably with DNA binding properties similar to G‐box binding factors found in, e.g. Arabidopsis thaliana. Stimulation with lipopolysaccharide, or overexpression of the mitogen‐activated protein kinase kinase kinase, MEKK‐1, further deepened the repression in primary cortical glial cells. It is suggested that this novel Jun binding repressor helps to control basal expression levels of CYP2E1, and modulates the response to inflammatory factors. Future in vivo experiments will, however, be required for a full appreciation of the role of this repressor in the complex regulation of CYP2E1 during inflammatory conditions.
Multi‐modal regulation of endogenous D1 dopamine receptor expression and function in the CAD catecholaminergic cell line
Authors: Pasuit Jennifer B. , Li Zhuting , Kuzhikandathil Eldo V.
Published: June 2004

Dopamine receptors exhibit tissue‐ and cell type‐specific expression that is modulated during development, aging and in diseases such as Parkinson's. The molecular mechanisms regulating expression of dopamine receptors are not well understood, in part due to the lack of a model cell line that not only expresses endogenous dopamine receptors but also has the requisite regulatory mechanisms. Here, we demonstrate that the CAD catecholaminergic cell line expresses D1, D2, D3 and D5 dopamine receptor subtypes and associated signaling proteins. CAD cell differentiation induced by serum withdrawal increases the levels of D1 receptor mRNA by transcriptional up‐regulation. This increase is also mimicked by the neurotrophin NT3. Interestingly, the increase of D1 receptor mRNA does not result in increased levels of D1 receptor protein in differentiated CAD cells. Furthermore, while the D1 receptor protein is expressed in differentiated CAD cells, it loses its ability to activate adenylyl cyclase. We demonstrate that the post‐transcriptional regulation is not due to decreased D1 receptor mRNA stability or generation of a truncated D1 receptor mRNA, and that the down‐regulation of D1 receptor function in differentiated CAD cells is mediated by post‐translational mechanisms that decrease cell surface receptor expression by altering receptor processing and trafficking.
MGLuR5 activation reduces β‐amyloid‐induced cell death in primary neuronal cultures and attenuates translocation of cytochrome c and apoptosis‐inducing factor
Authors: Movsesyan Vilen A. , Stoica Bogdan A. , Faden Alan I.
Published: June 2004

Activation of metabotropic glutamate receptor 5 (mGluR5) has been shown to reduce caspase‐dependent apoptosis in primary neuronal cultures induced by staurosporine and etoposide. β‐Amyloid (Aβ)‐induced neurotoxicity in culture appears to be in part caspase mediated. In the present studies the effects of treatment with an mGluR5 agonist or antagonist on Aβ‐induced neuronal apoptosis were examined in rat cortical neuronal cultures. Pretreatment with the selective mGluR5 agonist (RS)‐2‐chloro‐5‐hydroxyphenylglycine (CHPG) markedly reduced the number of apoptotic cells after exposure to Aβ (25–35), as well as associated LDH release. Blockade of mGluR5 by the selective antagonist, 2‐methyl‐6‐(phenylethynyl)pyridine (MPEP) attenuated these effects of CHPG. A similar neuroprotective effect of mGluR5 activation by CHPG was observed in cultures treated with full‐length Aβ peptide (1–42). CHPG attenuated Aβ (25–35)‐induced cytochrome c release and decreased levels of active caspase‐3 protein. CHPG also reduced translocation of apoptosis‐inducing factor (AIF) induced by Aβ (25–35). Thus, mGluR5 activation limits the release of mitochondrial proteins associated with induction of both caspase‐dependent and ‐independent apoptosis.
In vivo inhibition of Aβ production by memapsin 2 (β‐secretase) inhibitors
Authors: Chang Wan‐Pin , Koelsch Gerald , Wong Stephen , Downs Deborah , Da Huining , Weerasena Vajira , Gordon Brian , Devasamudram Thippeswamy , Bilcer Geoffrey , Ghosh Arun K. , Tang Jordan
Published: June 2004

We have previously reported structure‐based design of memapsin 2 (β‐secretase) inhibitors with high potency. Here we show that two such inhibitors covalently linked to a ‘carrier peptide’ penetrated the plasma membrane in cultured cells and inhibited the production of β‐amyloid (Aβ). Intraperitoneal injection of the conjugated inhibitors in transgenic Alzheimer's mice (Tg2576) resulted in a significant decrease of Aβ level in the plasma and brain. These observations verified that memapsin 2 is a therapeutic target for Aβ reduction and also establish that transgenic mice are suitable in vivo models for the study of memapsin 2 inhibition.
Focal dysfunction of the proteasome: a pathogenic factor in a mouse model of amyotrophic lateral sclerosis
Authors: Kabashi Edor , Agar Jeffrey N. , Taylor David M. , Minotti Sandra , Durham Heather D.
Published: June 2004

Mutations in the Cu/Zn‐superoxide dismutase (SOD‐1) gene are responsible for a familial form of amyotrophic lateral sclerosis (fALS). The present study demonstrated impaired proteasomal function in the lumbar spinal cord of transgenic mice expressing human SOD‐1 with the ALS‐causing mutation G93A (SOD‐1G93A) compared to non‐transgenic littermates (LM) and SOD‐1WT transgenic mice. Chymotrypsin‐like activity was decreased as early as 45 days of age. By 75 days, chymotrypsin‐, trypsin‐, and caspase‐like activities of the proteasome were impaired, at about 50% of control activity in lumbar spinal cord, but unchanged in thoracic spinal cord and liver. Both total and specific activities of the proteasome were reduced to a similar extent, indicating that a change in proteasome function, rather than a decrease in proteasome levels, had occurred. Similar decreases of total and specific activities of the proteasome were observed in NIH 3T3 cell lines expressing fALS mutants SOD‐1G93A and SOD‐1G41S, but not in SOD‐1WT controls. Although overall levels of proteasome were maintained in spinal cord of SOD‐1G93A transgenic mice, the level of 20S proteasome was substantially reduced in lumbar spinal motor neurons relative to the surrounding neuropil. It is concluded that impairment of the proteasome is an early event and contributes to ALS pathogenesis.
NMDA‐receptor regulation of muscarinic‐receptor stimulated inositol 1,4,5‐trisphosphate production and protein kinase C activation in single cerebellar granule neurons
Authors: Young Kenneth W. , Garro M. Asier , Challiss R. A. John , Nahorski Stefan R.
Published: June 2004

Inositol 1,4,5‐trisphosphate (InsP3) production in single cerebellar granule neurons (CGNs) grown in culture was measured using the PH domain of phospholipase C δ1 tagged with enhanced green fluorescent protein (eGFP‐PHPLCδ1). These measurements were correlated with changes in intracellular free Ca2+ determined by single cell imaging. In control CGNs, intracellular Ca2+ stores appeared replete. However, the refilling state of these stores appeared dependent on the fluorophore used to measure Ca2+‐release. Thus, methacholine (MCH), acting via muscarinic acetylcholine‐receptors (mAchRs), mobilised intracellular Ca2+ in cells loaded with fluo‐3 and fura‐4f, but not fura‐2. Confocal measurements of single CGNs expressing eGFP‐PHPLCδ1 demonstrated that MCH stimulated a robust peak increase in InsP3, which was followed by a sustained plateau phase of InsP3 production. In contrast, glutamate‐induced InsP3 signals were weak or not detectable. MCH‐stimulated InsP3 production was reduced by chelation of intracellular Ca2+ with BAPTA, and emptying of intracellular stores with thapsigargin, indicated a positive feedback effect of Ca2+ mobilisation onto PLC activity. In CGNs, NMDA‐ and KCl‐mediated Ca2+‐entry significantly enhanced MCH‐induced InsP3 production. Furthermore, mAchR‐mediated PLC activation appeared sensitive to the full dynamic range of intracellular Ca2+ increases stimulated by 100 μm NMDA. This dynamic regulation was also observed at the level of PKC activation indicated by an enhanced translocation of eGFP‐tagged myristoylated alanine‐rich C kinase substrate (MARCKS) protein in cells stimulated with MCH. Thus, NMDA‐mediated Ca2+ influx and PLC activation may represent a coincident‐detection system whereby ionotropic and metabotropic signals combine to stimulate InsP3 production and PKC‐mediated phosphorylation events in CGNs.
Netrin‐1 is crucial for the establishment of the dorsal column‐medial lemniscal system
Authors: Kubota Chikara , Nagano Takashi , Baba Hisatoshi , Sato Makoto
Published: June 2004

The dorsal column‐medial lemniscal system is a significant sensory pathway that mediates touch and limb position sense. In this system, axons from the second‐order neurons in the dorsal column nuclei form the internal arcuate fibers, cross the ventral midline (floor plate) within the medulla oblongata, and then project to the thalamus as the medial lemniscus. Here we demonstrate that Netrin‐1, which is secreted from the floor plate in the medulla oblongata, is indispensable to the formation of the dorsal column‐medial lemniscal system. Axons from the dorsal column nuclei cross the midline at around embryonic day 11 in mice. Concurrently, Netrin‐1 mRNA and its receptor DCC (deleted in colorectal cancer) were expressed in the floor plate and commissural axons there, respectively. In our explant culture experiments, the floor plates of the embryonic 11‐day‐old mutant Netrin‐1 homozygous mice did not attract axons from the dorsal column nuclei of ICR mice, while those from the wild type littermates did. Moreover, we observed that although the dorsal column nuclei developed in situ in mutant mice, their axons were not attracted toward the floor plate: they did not cross midline and remained ipsilaterally, without forming the internal arcuate fibers, in embryonic 17‐day‐old mutant Netrin‐1 homozygous mice.
Endocannabinoids accumulate in spinal cord of SOD1G93A transgenic mice
Authors: Witting Anke , Weydt Patrick , Hong Soyon , Kliot Michel , Mőller Thomas , Stella Nephi
Published: June 2004

Approximately 2% of amyotrophic lateral sclerosis (ALS) cases are caused by mutations in the super oxide dismutase 1 (SOD1) gene and transgenic mice for these mutations recapitulate many features of this devastating neurodegenerative disease. Here we show that the amount of anandamide (AEA) and 2‐arachidonoylglycerol (2‐AG), two endocannabinoids that have neuroprotective properties, increase in spinal cord of SOD1G93A transgenic mice. This increase occurs in the lumbar section of spinal cords, the first section to undergo neurodegeneration, and is significant before overt motor impairment. Our results show that chronic neurodegeneration induced by a genetic mutation increases endocannabinoid production possibly as part of an endogenous defense mechanism.
Prevalence of HFE mutations among the Thai population and correlation with iron loading in haemoglobin E disorder*
Authors: Viprakasit Vip , Vathesathokit Prin , Chinchang Worrawut , Tachavanich Kalaya , Pung‐Amritt Parichat , Wimhurst Victoria L. C. , Yenchitsomanus Pa‐Thai , Merryweather‐Clarke Alison T. , Tanphaichitr Voravarn S.
Published: June 2004

Abstract:  Co‐inheritance of HFE mutations has a substantial role in iron overload in β‐thalassaemia carriers in north European populations where two HFE mutations, C282Y and H63D, are prevalent. In Thailand, there was little information about the allele frequency of HFE mutations. It is of interest to determine whether such determinants represent a potential risk in developing iron overload as nearly 40% of the Thai population carry either one of thalassaemia or haemoglobinpathy alleles. A total of 380 normal controls from five different regions including Bangkok were screened for the HFE C282Y, H63D and IVS5+1 G→A alleles. In addition, 70 individuals with homozygous haemoglobin E (Hb EE) were also tested and their genotypes were correlated with levels of serum ferritin. H63D is the major HFE mutation found in the Thai population with an average allele frequency of 3% (range 1–5%). One individual was heterozygous for the splice site mutation IVS5 + 1 G → A, and the C282Y allele was not detected. In the Hb EE group, five individuals had iron deficiency (ferritin <12 μg/L) and the remaining 65 individuals had a wide range of serum ferritin levels of 16–700 μg/L. Four individuals with Hb EE were heterozygous for the H63D allele. No significant difference in serum ferritin level was detected in this group with or without the HFE mutation (137.2 ± 78 vs. 116.3 ± 128 μg/L). HFE mutations are relatively uncommon among the Thai population, and the average allele frequency of the ancient H63D mutation is similar to that of other countries in this region. Because of their paucity, it appears that these alleles are less likely to be responsible for high ferritin levels and iron loading in individuals with Hb E related disorders.
Different efficacy of mycophenolate mofetil as salvage treatment for acute and chronic GVHD after allogeneic stem cell transplant
Authors: Kim Jong Gwang , Sohn Sang Kyun , Kim Dong Hwan , Lee Nan Young , Suh Jang Soo , Lee Kun Soo , Lee Kyu Bo
Published: June 2004

Abstract: Objective: The therapeutic options currently available for treating refractory graft‐vs.‐host disease (GVHD) are limited. Therefore, the present study evaluated the efficacy of mycophenolate mofetil (MMF) as a salvage treatment for acute and chronic GVHD in allograft patients.

Methods: Twenty‐six consecutive patients with refractory acute (13 patients) or chronic (13 patients) GVHD were enrolled. The first‐line treatment for all patients with acute GVHD consisted of a combination of cyclosporine A (CyA) and steroids, while the first‐line treatment for chronic GVHD was steroids with or without CyA according to the risk group. MMF was added at a dose of 1.5 or 2 g daily and the steroids were tapered in the refractory cases.

Results: Four (30.8%) of 13 patients with refractory acute GVHD responded to MMF. When analyzing the overall results according to the type of acute GVHD, improvement was observed in four (30.8%) of 13 skin cases, four (44.4%) of nine liver cases, and two (22.9%) of nine gut cases. Ten (76.9%) of 13 patients with refractory chronic GVHD responded to MMF. The common side effects were gastrointestinal disturbance (26.9%) or infectious complications (23.1%). The estimated 2‐yr survival rate for patients with acute GVHD and chronic GVHD was 33.3% and 53.9%, respectively.

Conclusions: MMF would appear to be effective and safe for treating refractory chronic GVHD, yet not as effective for treating refractory acute gut GVHD. Accordingly, a prospective randomized clinical trial is warranted to assess the impact of MMF in the treatment of refractory GVHD.
Hematologic and molecular spontaneous remission following sepsis in acute monoblastic leukemia with translocation (9;11): a case report and review of the literature
Authors: Müller Claudia I. , Trepel Martin , Kunzmann Regina , Lais Angela , Engelhardt Rupert , Lübbert Michael
Published: June 2004

Abstract:  Spontaneous remission in patients with acute myeloid leukemia (AML) is a rarely reported phenomenon of usually short duration. The etiology remains unclear, but an association with preceding blood transfusions or bacterial infections has been reported. Triggered immune responses are suggested to play a potential role in the development of spontaneous remission. Acute monocytic leukemia was diagnosed in a 61‐yr‐old male patient. Cytogenetic analysis revealed a sole translocation (9;11) (q22;q23) and RT‐PCR the MLL/AF9 fusion gene. As a result of the patient's reduced performance status and septic condition, cytostatic therapy was withheld. No microorganisms could be detected. Hematologic and molecular remission occurred after initiating antibiotic therapy without any cytostatic treatment; 29 months after the initial diagnosis, he is in complete remission, and excellent physical condition. Our report includes a review of the literature since 1985, reporting cases of patients with AML and spontaneous remission together with informative cytogenetics. Balanced translocations such as in core binding factor (CBF) leukemias appear somewhat overrepresented. We speculate that AML‐specific T cells might be relevant for induction of spontaneous remission and need to be further investigated.
Fulminant bronchiolitis obliterans organizing pneumonia following 2 d of treatment with hydroxyurea, interferon‐α and oral cytarabine ocfosfate for chronic myelogenous leukemia
Authors: Kalambokis Georgios , Stefanou Dimitrios , Arkoumani Evanthia , Kitsanou Margarita , Bourantas Konstantinos , V. Tsianos Epameinondas
Published: June 2004

Abstract:  A 65‐yr‐old man developed increasing dyspnea and fulminant respiratory failure 48 h after introduction of hydroxyurea, oral cytarabine ocfosfate (YNK01) and interferon‐α for treatment of Philadelphia chromosome‐positive chronic myelogenous leukemia. The chest radiograph showed bilateral patchy infiltrates while computed tomography revealed multiple bullas, ground glass opacities, and patchy consolidations with possible cavitation. Bronchoscopic examination was normal and microbiological tests performed on all biologic fluids were negative. The patient did not respond to multiple antibiotic treatment and corticosteroid administration and died of progressive respiratory failure 5 d after chemotherapy introduction. The postmortem lung examination was consistent with the diagnosis of bronchiolitis obliterans organizing pneumonia (BOOP).
Safety and efficacy of caspofungin and liposomal amphotericin B, followed by voriconazole in young patients affected by refractory invasive mycosis
Authors: Cesaro Simone , Toffolutti Tiziana , Messina Chiara , Calore Elisabetta , Alaggio Rita , Cusinato Riccardo , Pillon Marta , Zanesco Luigi
Published: June 2004

Abstract:  Objective: Data on the use of combination of liposomal amphotericin B and caspofungin followed by voriconazole, as maintenance or further rescue treatment, in 10 patients with invasive mycosis are reported. Material and methods: The diagnoses were acute leukemia (7), myelodysplastic syndrome (1) and Hodgkin's lymphoma (1). All patients developed an invasive mycosis (proven, 3; probable, 6; and possible, 1) refractory to first‐line antifungal treatment (liposomal amphotericin B in all patients except one who received fluconazole). Results: Rescue therapy with a combination of caspofungin and liposomal amphotericin B was well tolerated, hypokalemia, and thrombophlebitis being the most common side‐effects. Combination therapy was administered for a median of 17 d, range 6–40. Among the nine patients with proven or probable mycosis, one was not evaluated because of early death caused by massive hemoptysis whilst in the remaining eight patients, the response was classified as complete, stable and failure in four, three, and one patients, respectively. Complete response was also observed in patient with possible mycosis. Eight of nine patients received voriconazole for a median of 75 d, range 42–194. Voriconazole was well tolerated although some drug interactions were observed during treatment with methotrexate and digoxin. After a median follow‐up of 125 d, nine of 10 patients are alive. Overall, a favorable response to antifungal treatment (including the case of possible mycosis) was obtained in eight of 10 patients. Conclusion: These data suggest that medical antifungal treatment may be intensified in severely ill patients without significantly compromising patient safety. The combination of synergistic antifungal drugs as well as their sequential use warrants further investigation by a larger randomized controlled study.
Disease‐ or therapy‐related bone marrow damage cannot be overcome by changes in stem cell source or dose in allogeneic transplantation
Authors: Novotny J. R. , Rosenthal C. , Elmaagacli A. H. , Dürig J. , Beelen D. W. , Dührsen U.
Published: June 2004

Abstract:  Objective:  To test whether the functional impairment of the host bone marrow (BM) microenvironment pre‐existing at the time of transplantation could be overcome by the increased content of immature cells in allogeneic peripheral blood stem cell transplantation (PBSCT) when compared with bone marrow transplantation (BMT).

Methods:  Cobble stone area forming cells (CAFC) were assayed in normal BM and BM after allogeneic BMT and PBSCT after stable engraftment. Groups were compared by two‐tailed t‐test.

Results:  While BM from 11 normal controls contained an average of 778.8 CAFC‐d35 per 106 low density bone marrow cells (LDBMC, range 453–1231 per 106 LDBMC), BM from patients after BMT contained an average of 123.7 CAFC‐d35 per 106 LDBMC (range 38–257) per 106 LDBMC. BM from patients transplanted with PBSC after myeloablative conditioning contained 128.3 (range 46–305) CAFC‐d35 per 106 LDBMC (P = 0.89 compared with BMT). Similar results were obtained when patients after PBSCT with non‐myeloablative conditioning were included (P = 0.62 compared with BMT). CAFC numbers in patients transplanted in early stages of myeloid leukaemia (acute myeloid leukaemia first remission, chronic myeloid leukaemia first chronic phase) were significantly higher than CAFC numbers in patients transplanted in more advanced stages (P = 0.008) or myelodysplastic syndrome (P = 0.023). The lowest CAFC numbers were found in two cases of retransplantation.

Conclusion:  Our findings indicate that the functional state of the BM microenvironment rather than stem cell dose or source is limiting for the homing and engraftment of immature haemopoietic cells in clinical transplantation.
Intracellular IFN‐γ expression by CD3+/CD8+ cell subset in B‐CLL patients correlates with stage of the disease
Authors: Podhorecka Monika , Dmoszynska Anna , Rolinski Jacek
Published: June 2004

Abstract:  Changes in the cytokine network may be responsible for malignant cell accumulation in B‐cell chronic lymphocytic leukaemia (B‐CLL). Among different cytokines of question interferon gamma (IFN‐γ) is indicated to prevent malignant cells from entering apoptosis. The aim of the study was to determine IFN‐γ production capacity of T‐cell subsets and B lymphocytes in B‐CLL patients in comparison with healthy individuals and during disease progression. Forty patients with newly diagnosed, untreated B‐CLL and 20 healthy individuals were studied. The two‐ and three‐colour flow cytometry techniques were used to detect intracellular cytokine expression. We detected statistically significantly higher percentage of both CD3+/CD4+/IFN‐γ+ and CD3+/CD8+/IFN‐γ+ in patients than in controls (P < 0.001 in both cases). Moreover the percentage of CD3+/CD8+/IFN‐γ+ cells correlated with stage of the disease (R = 0.39, P = 0.01) and parameters of disease progression like lymphocyte count and total tumour mass score (R = 0.33, P = 0.03 and R = 0.31, P = 0.04, respectively). By contrast, the percentage of CD19+/IFN‐γ+ cells in B‐CLL group was lower than in controls (P < 0.01). These findings indicate that T‐cell populations rather than malignant B cells are the source of IFN‐γ in B‐CLL patients. The subset of CD3+/CD8+ cells expressing IFN‐γ seems to play a special role in the disease progression and more precise investigation should elucidate its role as a prognostic marker in B‐CLL and a target for therapeutic strategies.
Excessive production of tumor necrosis factor‐alpha by bone marrow T lymphocytes is essential in causing bone marrow failure in patients with aplastic anemia
Authors: Hara Takeshi , Ando Kazuki , Tsurumi Hisashi , Moriwaki Hisataka
Published: June 2004

Abstract:  Aplastic anemia (AA) is regarded as an immunological disorder because of the clinical effect of immunosuppressive therapy. Recent studies have reported that cytokines play an important role in the development of AA. In the present study, we measured levels of T‐cell derived intracellular cytokine production in peripheral blood and bone marrow (BM) of patients with AA. We demonstrated that BM lymphocytes, particularly CD4+ and CD8+ T cells, in patients with AA produced significantly higher amounts of tumor necrosis factor‐alpha (TNF‐α), compared with lymphocytes in normal controls. We have previously reported that expression of TNF receptor (R)1 and TNFR2 in the CD34+ CD38− and CD34+ CD38+ fractions of patients with AA is significantly higher than those in normal control. These results indicate that BM stem cells in patients with AA may possess high sensitivity to TNF‐α. This in turn suggests that TNF‐α affects hematopoiesis at an earlier stage in AA patients than in normal controls. We strongly support the hypothesis that a simultaneous increase in TNF‐α production by BM lymphocytes and sensitivity of stem cells to TNF‐α leads to BM failure in AA.
WT1 gene expression: useful marker for minimal residual disease in childhood myelodysplastic syndromes and juvenile myelo‐monocytic leukemia?
Authors: Bader Peter , Niemeyer Charlotte , Weber Gerrit , Coliva Tiziana , Rossi Vincenzo , Kreyenberg Hermann , Gerecke Anja , Biondi Andrea
Published: June 2004

Abstract:  The WT1 gene is considered to be highly expressed in patients with acute myeloid leukemia (AML), acute lymphoblastic leukemia and chronic myeloid leukemia and is thought to play a key role in maintaining the viability of leukemia cells. However, little is known about the WT1 gene expression levels in pediatric patients with juvenile myelo‐monocytic leukemia (JMML) and myelodysplastic syndromes (MDS). We studied WT1 expression in diagnostic bone marrow (BM) and peripheral blood (PB) samples of 90 patients with JMML, low grade MDS, advanced MDS and myelodysplasia‐related AML in BM (n = 20) and PB (n = 18) samples of normal healthy volunteer donors.
Matrix metalloproteinase inhibitor reduces apoptosis induction of bone marrow cells in MDS‐RA
Authors: Arimura Kosei , Arima Naomichi , Matsushita Kakushi , Ohtsubo Hideo , Fujiwara Hiroshi , Kukita Toshimasa , Ozaki Atsuo , Hagiwara Takafumi , Hamada Heiichiro , Yoshino Kohichiro , Tei Chuwa
Published: June 2004

Abstract:  Background and objectives: We examined the involvement of apoptosis with myelodysplastic syndrome (MDS) accompanied by peripheral cytopenias despite normo‐hypercellular bone marrow. Materials and methods: Bone marrow smears from 31 patients with MDS‐refractory anemia (RA) and five normal controls were stained using the in situ end labeling (ISEL) method. Next, the inhibitory effects of a caspase‐3 inhibitor, matrix metalloproteinase inhibitor (MMPI), anti‐tumor necrosis factor (TNF)‐α or anti‐Fas antibody upon the apoptosis induction in overnight cultures of bone marrow cells from the patients were examined. Further, TNF‐α, transforming growth factor (TGF)‐β and soluble Fas ligand (sFasL) concentrations in culture supernatants of the cells were assessed by enzyme‐linked immunosorbent assay (ELISA). Results: The incidence of ISEL‐positive cells among MDS patients was significantly higher than in normal controls (50.8 ± 14.0% vs. 11.3 ± 2.4%; P < 0.0001). A caspase‐3 inhibitor reduced significantly the ISEL‐positive rates (32.6 ± 15.2% vs. 50.2 ± 16.5%; P < 0.0001). Anti‐TNF‐α or anti‐Fas antibody reduced the ISEL‐positive rates significantly (28.2 ± 6.0%, 29.2 ± 5.8%, vs. 44.2 ± 3.4%, P < 0.001, P = 0.001, respectively). KB‐R7785 also significantly decreased the ISEL‐positive rates (18.0 ± 9.3% vs. 43.6 ± 14.0%; P < 0.0001). The concentration of TNF‐α was significantly reduced by KB‐R7785 (P < 0.05), whereas that of TGF‐β was not. Concentration of sFasL was under detectable level in the present assay system. The derivatives of KB‐R7785 that can be administrated orally showed inhibitory effect on apoptosis induction as well. Conclusions: These findings suggest that MMPIs inhibits the apoptosis induction of MDS bone marrow cells via the inhibition of TNF‐α and probably sFasL secretion, and that MMPIs can be used to control the abnormal induction of apoptosis in MDS.
Serum angiogenin is not elevated in patients with early B‐cell chronic lymphocytic leukemia but is prognostic factor for disease progression
Authors: Molica Stefano , Vitelli Gaetano , Levato Domenico , Giannarelli Diana , Vacca Angelo , Cuneo Antonio , Ribatti Domenico , Digiesi Giovanna
Published: June 2004

Abstract:  The association between angiogenin and cancer progression and poor outcome in solid tumors has been documented, but its significance in leukemias has not been evaluated. Using an ELISA technique (Quantikine Human Angiogenin Immunoassay; R&D Systems), we measured serum angiogenin levels in 77 previously untreated Binet stage A B‐cell chronic lymphocytic leukemia (CLL) patients. No difference in angiogenin serum levels could be found between patients (median: 295 ng/mL; range: 74–1700) and 15 age‐ and sex‐matched healthy controls (median: 264 ng/mL; range: 29–1835) (P = NS; Mann–Whitney test). Increased angiogenin serum level was associated with higher LDH (P = 0.03) and β2‐m (P = 0.007) concentrations. However, angiogenin did not reflect the extent of bone marrow (BM) angiogenesis as evaluated by microvessel area (P = 0.611), circulating levels of vascular endothelial growth factor (VEGF) (P = 0.873) and basic fibroblastic growth factor (FGF‐2) (P = 0.421). When the 25 patients with available data were stratified into the four major cytogenetic categories (normal karyotype, 13q as a sole aberration, 12q trisomy, 11q or 17p deletion) and aberrations were compared with angiogenin serum levels, no correlation was found (P = 0.651; Kruskall–Wallis test). A cut‐off of angiogenin serum level corresponding to median (i.e. 330 ng/mL) or higher identified later upstaging and longer progression‐free survival (PFS). The 5‐yr PFS was 51.5% for patients with angiogenin levels lower than median and 85% for patients with higher values [P = 0.03; hazard ratio (HR) = 2.86; 95% CI: 1.08–6.72]. Although in multivariate analysis only Rai substages (P = 0.00001) and peripheral blood lymphocytosis (P = 0.009) retained their prognostic significance, angiogenin could be incorporated into the Rai substages thus leading to the identification of the following risk categories: (i) stage 0 (angionenin >330 ng/mL); (ii) stage 0 (angiogenin <330 ng/mL) + stage I–II (angiogenin >330 ng/mL); and (iii) stage I–II (angiogenin <330 ng/mL). The 40‐month PFS were as follows: 85%, 65%, 25% (χ2 for trend = 6.33; d.f. = 1; P = 0.01). In conclusion, serum angiogenin levels although not increased in comparison with healthy controls, may predict clinical outcome of patients with early CLL and help to refine Rai's stratification.
The application of North American CT scan criteria to an Australian population with minor head injury
Authors: Rosengren David , Rothwell Sean , Brown Anthony FT , Chu Kevin
Published: June 2004

Objective:  To determine potential changes in the number of CT head scans performed if the New Orleans Criteria (NOC) or Canadian CT Head Rule (CCTR) was applied to an Australian emergency department population of minor head injured (MHI) patients.

Methods:  A retrospective chart review was conducted in an adult metropolitan teaching hospital in Brisbane. All patients presenting over a 3‐month period with a GCS Score of 15 following an MHI and had a CT head scan performed were selected for analysis. Using clinically significant CT abnormalities and neurological intervention as the outcome measures, the NOC and CCTR were applied to determine if CT scanning was considered necessary.

Results:  Of the 240 patients reviewed, 230 had a normal CT scan and 10 had clinically significant CT abnormalities. One patient with CT abnormality required neurosurgical intervention. Application of the NOC would have resulted in a 3.8% (95% CI 1.7–7.0%) reduction in CT scans performed without missing any patients with CT abnormalities or requiring neurological intervention. Application of the CCTR using both high and low risk factors would have resulted in a 46.7% (95% CI 40.2–53.2%) reduction in CT scans performed without missing the patient requiring neurological intervention, but would not have detected two patients with clinically significant CT abnormalities.

Conclusion:  Neither the NOC nor the CCRT appear suitable for significantly reducing the number of normal CT head scans performed without missing clinically significant CT abnormalities when applied to our current clinical practice.
Wheeze and mediastinal mass: A challenging patient
Authors: Heinz Peter , Dunne John
Published: June 2004

Airway obstruction is a recognized complication in children with mediastinal masses. They typically present with difficulty in breathing and associated respiratory noises. General anaesthesia in these patients can lead to complete airway obstruction with fatal consequences. Successful management in the ED necessitates rapid recognition of the underlying problem and appropriate intervention. We report the case of a 7‐year‐old boy presenting with respiratory collapse and describe the management that led to successful resuscitation.
Extensive surgical emphysema following restorative dental treatment
Authors: Aquilina Peter , McKellar Geoffrey
Published: June 2004

We present a patient with extensive surgical emphysema following the dental restoration the upper left first molar (tooth 26) with a high speed turbine handpiece. The clinical findings and management of subcutaneous cervical emphysema are discussed.
Not just gastroenteritis: Thyroid storm unmasked
Authors: Charles Rabind Antony , Goh Su‐Yen
Published: June 2004

Establishing the diagnosis of thyroid storm is difficult in the ED, especially where there is no antecedent history of thyroid disease or clinical clues like goitre, exophthalmos or altered mentation, yet early recognition and treatment are essential in reducing mortality and morbidity from this endocrine emergency. We present a case where suspected infective gastroenteritis in a newly diagnosed diabetic masked the major symptomatology of thyroid storm, and review the diagnosis and management of thyrotoxic crisis.
A casemix model for estimating the impact of hospital access block on the emergency department
Authors: Stuart Peter
Published: June 2004

Objective: To determine the ED activity and costs resulting from access block.

Methods: A casemix model (AWOOS) was developed to measure activity due to access block. Using data from four hospitals between 1998 and 2002, ED activity was measured using the urgency and disposition group (UDG) casemix model and the AWOOS model with the purpose of determining the change in ED activity due to access block.

Results: Whilst the mean length of stay in ED (admitted patients) increased by 93% between 1998 and 2002, mean UDG activity increased by 0.63% compared to a mean increase in AWOOS activity of 24.5%. The 23.9% difference between UDG and AWOOS activity represents the (unmeasured) increase in ED activity and costs for the period 1998–2002 resulting from access block.

Conclusion: The UDG system significantly underestimates the activity in EDs experiencing marked access block.
Intermediate dose metoclopramide is not more effective than standard dose metoclopramide for patients who present to the emergency department with nausea and vomiting: A pilot study
Authors: Cham Swee , Basire Mary , Kelly Anne‐Maree
Published: June 2004

Objective:  To determine whether intermediate dose metoclopramide is more effective than standard dose metoclopramide for patients who present to the ED with nausea and vomiting.

Methods:  This prospective, single‐blind, randomized trial was conducted in the ED of two community teaching hospitals in a convenience sample of adult patients who presented to ED with nausea and vomiting. Patients were randomized to receive either 10 mg metoclopramide IV or 0.4 mg/kg IV (up to a maximum dose of 32 mg). The outcomes measured were: change in nausea score (measured on a 11‐point verbal rating scale), requirement for other anti‐emetic drug administration, and presence of side‐effects.

Results:  Fifty‐eight patients were eligible for analysis (34 in 10 mg group and 24 in 0.4 mg/kg group). Median reduction in nausea score in 10 mg group was four (range 0–10, 95% CI 3–5) compared with five for 0.4 mg/kg group (range −1–10, 95% CI 4–6). This difference was not statistically significant (P = 0.629). Five patients in the 10 mg group required rescue anti‐emetic, compared with three in the 0.4 mg/kg group (P = 1.00). There were no side‐effects in the 10 mg group and two in the 0.4 mg/kg group.

Conclusion:  This study suggests that there is no difference in effectiveness between 10 mg and 0.4 mg/kg of metoclopramide in the ED population with nausea and vomiting.