bluuebunny/crossref_metadata_2025 huggingface dataset:
Investigation of the Prognostic Value of TNF-α Gene Polymorphism among Patients Treated with Infliximab, and the Effects of Infliximab Therapy on TNF-α Production and Apoptosis
Authors: Balog Attila , Klausz Gergely , Gál János , Molnár Tamás , Nagy Ferenc , Ocsovszky Imre , Gyulai Zsófia , Mándi Yvette
Published: October 2004

Objectives: Infliximab, a chimeric anti-tumor necrosis factor (TNF) antibody, is highly effective for the treatment of Crohn’s disease (CD) and rheumatoid arthritis (RA). Our experiments focused on RA and CD patients receiving infliximab. Since cytokine production is largely determined by genetic factors, the promoter polymorphisms of TNF-α were examined among these patients. Additionally, the changes caused by infliximab in the TNF-α-producing ability and apoptosis of peripheral blood mononuclear cells (PBMCs) were investigated. Methods: The TNF-α genotypes were analyzed by PCR-RFLP. The in vitro TNF-α production of the PBMCs was detected by flow cytometric analysis. The TNF-α concentration in the supernatant was measured by bioassay. Apoptosis was detected by annexin V-fluorescein isothiocyanate labeling. Results and Conclusions: 8 of the 12 nonresponder patients carried the TNF A allele associated with high TNF-α production. We suggest that the determination of TNF polymorphism may help identify more suitable candidates for infliximab treatment. Although in vitro infliximab treatment for 48 h resulted in significant (44.2 ± 1.17%) apoptosis in PBMCs, in ex vivo samples from RA patients who received infliximab, apoptosis was only 13.3 ± 1.6%. Furthermore, infliximab did not result in irreversible inhibition of the TNF-α-producing ability or in the significant apoptosis of PBMCs.
Expression of c-myc mRNA in Hepatocellular Carcinomas, Noncancerous Livers, and Normal Livers
Authors: Ikeguchi Masahide , Hirooka Yasuaki
Published: October 2004

Objectives: c-myc is a transcription factor that regulates cell proliferation, differentiation, and apoptosis. However, little is known about the clinical importance of c-myc mRNA expression in a primary hepatocellular carcinoma (HCC). In this study, we investigated whether a correlation existed between the level of c-myc mRNA expression and the occurrence of apoptosis or proliferative activity in an HCC. Methods: The levels of c-myc mRNA expression were quantified by real-time reverse transcriptase polymerase chain reaction (RT-PCR) and compared with immunohistochemical detection of the occurrence of spontaneous apoptosis and proliferative activity of cancer cells in 59 surgically resected HCCs. Results: The mean level of c-myc mRNA expression in 59 HCCs (1.19) was significantly suppressed compared with that in 59 noncancerous liver tissues (1.44, p < 0.0001). The level of c-myc mRNA expression was not correlated with tumor differentiation, tumor progression, proliferative activity of cancer cells, or patient survival. However, there was a significant positive correlation between the levels of c-myc mRNA expression and the occurrence of apoptosis in 59 HCCs (p < 0.001). Conclusions: Inhibition of c-myc mRNA expression may obstruct the induction of apoptosis of HCC cells and lead to uncontrolled cell growth.
Role of Potassium Channels in the Frequency-Dependent Activity of Regenerating Nerves
Authors: Mert Tufan , Gunay Ismail , Daglioglu Yusuf Kenan
Published: October 2004

After a peripheral nerve injury, ion channel organization and the electrical properties of nerve fibers drastically change during the regeneration process. The present study was designed to compare the frequency-dependent characteristics of regenerating nerves in the presence of 4-aminopyridine (4-AP) and tetraethylammonium (TEA). The results showed that increasing the stimulus frequency produced a greater impulse blockade (frequency-dependent block – FDB) and distinct hyperpolarizing afterpotentials (HAPs) in regenerating nerves. In particular, regenerating sciatic nerves 15 days post-crush (dpc) were more sensitive to the frequency-dependent stimulations than 38-dpc and intact nerves in the presence or absence of drugs. The frequency-dependent effects of TEA on the compound action potentials (CAPs) appeared when TEA was applied to 4-AP-treated nerves. This shows that TEA-sensitive channels may not be masked by the myelin. 4-AP was here found to have more pronounced frequency-dependent effects on regenerating nerves than on intact nerves. Delayed depolarization (in 38-dpc: 22.6 ± 1.3 mV and 47.52 ± 3.63 ms, in intact: 12.0 ± 1.9 mV and 88.51 ± 4.72 ms) elicited by 4-AP resulted in an increase in FDBs and HAP amplitudes. These results suggest that 4-AP-sensitive channels may play important roles in frequency-dependent nerve conduction. Consequently, regenerating or myelin damaged nerves are more sensitive to repetitive firing with or without drug. An understanding of the frequency-dependent properties of regenerating nerves may be of value in the treatment of the nerve diseases.
Effects of Modulation of the NO/cGMP Pathway in Tumor Cell Lines Derived from the Upper Airway Tract
Authors: Papapetropoulos Nektarios , Zhou Zongmin , Roussos Charis , Goumas Panagiotis , Papapetropoulos Andreas
Published: October 2004

Nitric oxide (NO) is known to be expressed in a variety of cell types and exert its effects through autocrine and paracrine mechanisms. To characterize the NO/cGMP pathway in tumor cells of the upper airway tract, we studied the cell lines Detroit 562, FaDu and FAT7. Using isoform-specific antibodies, we were unable to detect expression of NO synthases in the above-mentioned cells lines. To evaluate whether tumor cells respond to NO, we exposed cells to the NO donor sodium nitroprusside (SNP). Stimulation of Detroit 562 and FaDu with SNP (10 µmol/l to 1 mmol/l) led to a concentration-dependent increase in cGMP accumulation. In addition, incubation of cells with SNP, but not 8 Br-cGMP, reduced Detroit 562 cell number. As exposure of cells to SNP decreased 3H-thymidine incorporation without inducing DNA fragmentation, we attributed the observed decrease in cell number to inhibition of cell proliferation rather than induction of apoptosis. On the other hand, exposure of Detroit 562 to high concentrations of SNP (1 mmol/l) led to apoptosis and increased the release of vascular endothelial growth factor. We conclude that, although human cell lines derived from the upper airway tract do not produce NO, they respond to NO released by neighboring cells and that exposure to NO exerts an anti-proliferative/apoptotic effect that is independent of cGMP generation.
Comparison of the Inhibitory Efficacy of Four Belladonna Drugs on Gastrointestinal Movement and Cognitive Function in Food-Deprived Mice
Authors: Pan S.Y. , Han Y.F.
Published: October 2004

Belladonna drugs, scopolamine (Sco), atropine (Atr), anisodine (AT3), and anisodamine (Ani), frequently used for gastrointestinal motility disorders often produce adverse effects on the central nervous system. In the present work, these drugs (0.05, 0.5, 5, and 50 µmol kg–1, i.p.) were evaluated for their potential to inhibit gastrointestinal motility and cognition in mice. Results showed that the maximum inhibitory rates of Sco, Atr, AT3, and Ani on gastric emptying and small intestinal movement were 29.78, 40.69, 12.30, and 17.99% and 51.98, 58.46, 46.51, and 46.22%, respectively. The affinities of Sco, Atr, AT3, and Ani for muscarinic receptors in the whole mice were 1.62, 1.48, 2.28, and 1.11 µmol kg–1 for the stomach or 0.30, 1.12, 0.59, and 1.14 µmol kg–1 for the small intestine, respectively. The minimal effective doses for impairing avoidance-response learning were 5 µmol kg–1 for Sco, Atr, or AT3 and 50 µmol kg–1 for Ani. The initial doses for insulting the avoidance-response memory or open-field memory were 0.5, 5, 5, and 50 µmol kg–1 or 5, 5, 5, and >50 µmol kg–1 for Sco, Atr, AT3, and Ani, respectively. We conclude that the relative susceptibility of the mouse’s tissue or function capacities towards the inhibitory effects of belladonna drugs is small intestine > stomach > avoidance-response memory > avoidance-response learning > open-field memory.
Treatment with Buserelin, an Agonist of Gonadotropin-Releasing Hormone, Suppresses Ovarian Hyperstimulation Syndrome Induced in Rabbits
Authors: Oshima K. , Suzuki K. , Makita S. , Yoshida Y.
Published: October 2004

Human menopausal gonadotropin (hMG, 75 IU/body/day) and a gonadotropin-releasing hormone (GnRH) agonist buserelin (1, 10, 100 µg/kg/day) were simultaneously administered to female rabbits by the subcutaneous route for 7 days, and the effects on organ weights, plasma hormones and weight of ascitic fluid were examined. Treatment with hMG increased the ovarian weight, plasma estradiol and weight of ascites, thus indicating that ovarian hyperstimulation syndrome had been induced. Simultaneous treatment with buserelin decreased the changes induced by hMG. GnRH agonists can thus be surmised to reduce the severity of ovarian hyperstimulation syndrome in the rabbit. However, caution is needed when extrapolating the results of this rabbit model to humans.
Modulatory Effect of the PDE-5 Inhibitor Sildenafil in Diabetic Neuropathy
Authors: Patil Chandrashekhar S. , Singh Vijay Pal , Singh Sukhjeet , Kulkarni Shrinivas K.
Published: October 2004

Diabetic neuropathy is one of the most frequent peripheral neuropathies associated with hyperalgesia and hyperesthesia. Besides alteration in the levels of neurotransmitter, alteration in the neuronal nitric oxide synthase (nNOS) is a key factor in the pathogenesis of diabetic neuropathy. The present study was aimed at evaluating the role of PDE-5 inhibitor on nociception in streptozotocin-induced diabetes in animal models of nociception (writhing assay in mice and paw hyperalgesia test in rats). Diabetic animals showed a significant decrease in pain threshold as compared to non-diabetic animals in both tests, indicating diabetes induced hyperalgesia in mice and rats. The PDE-5 inhibitor, sildenafil, significantly increased the pain threshold in both diabetic and non-diabetic animals. However, L-NAME, a non-specific NOS inhibitor and methylene blue (MB), a guanylate cyclase inhibitor blocked the antinociceptive effect. The per se administration of L-NAME or MB augmented the hyperalgesic response in diabetic animals with little or no effect in non-diabetic animals, indicating the alteration of NO-cGMP pathway in diabetes. The results in the present study demonstrate that the decreased nNOS-cGMP system may play a crucial role in the pathogenesis of diabetic neuropathy.
Possible Role of Dexamethasone in Sensitizing the Beta-2-Adrenergic Receptor System in vivo in Calves during Concomitant Treatment with Clenbuterol
Authors: Abraham G. , Gottschalk J. , Ungemach F. R.
Published: October 2004

β2-Agonists blunt the function of the β-adrenoceptor G-protein adenylate cyclase-signalling system, whereas glucocorticoids reverse the agonist-mediated diminished β-adrenergic responses; however, these effects have not been reported in vivo in calf lymphocytes. In this study, we first investigated the presence of the β2-adrenergic receptors on calf lymphocytes, and second we tested the effects of either clenbuterol alone or in combination with dexamethasone on receptor expression and function (isoproterenol-induced intracellular adenosine 3′,5′-cyclic monophosphate (cAMP) formation) in vivo. (–)-[125I]-Iodocyanopindolol (ICYP) binding to intact calf lymphocytes was rapid, saturable (maximal number of binding sites 987 ± 89 ICYP-binding sites/cell, n = 4) and of high affinity (KD value 17.23 ± 2.8 pmol/l, n = 4). These binding sites were of the β2-subtypes of adrenoceptors as indicated by the fact that β-agonists inhibited ICYP binding with an order of potency: (–)-isoproterenol > (–)-adrenaline > (–)-noradrenaline. Furthermore, the selective β2-adrenoceptor antagonist ICI 118.551 was about >1,500 times more potent in inhibiting ICYP binding than was the β1-selective adrenoceptor antagonist CGP 20712A. Consequently, calves were treated with clenbuterol (1.0 µg/kg b.i.d., i.v.) for 9 days alone or simultaneously with dexamethasone (0.1 mg/kg, i.v., once a day for 4 days). Clenbuterol decreased the number of lymphocyte β2-adrenergic receptors by about 40–50% after only 48 h of drug administration. This was accompanied by a decrement in isoproterenol-induced lymphocyte cAMP formation. Upon application of both drugs, dexamethasone restored the clenbuterol-mediated decrease in β2-adrenoceptors and cAMP production. Dexamethasone elevated the number of β2-adrenoceptors and cAMP almost 1.5- to 2-fold at 24 h of drug administration, an effect that persisted for up to 24 h following drug withdrawal. Neither clenbuterol nor the combination with dexamethasone had an influence on the affinity of the receptor for the ligand. The present results demonstrate that dexamethasone in vivo upregulates the number and function of calf lymphocyte β2-adrenoceptors, and thus enhances the sensitivity of the β2-adrenoceptor signal-transduction pathway for clenbuterol during concomitant treatment with both drugs.
Method for Selecting Populations of Rat Articular Chondrocytes That Exhibit Distinct Growth and Metabolic Characteristics, and Their Responses to Growth Factors, PMA and Vitamin D3
Authors: Panasyuk Andrei , Khatib Abdel-Majid , Colantuoni Gladys , Quintero Maritza , Lomri Abderrahim , Mitrovic Dragoslav R.
Published: October 2004

Chondrocytes were released from articular cartilage fragments of 6-week-old Wistar rats by a 2-hour treatment with bacterial collagenase. The cells from one animal were seeded in a 25-cm2 culture flask at a density of 105 cells/cm2. After 1 h, the flask was gently shaken and the medium, containing nonadherent cells, was transferred to a new flask. The attached cells were incubated with 5 ml of fresh medium. This procedure was repeated after 3, 24, 48 and 96 h. Resulting cell populations were then analyzed. The earlier cells attached, the more rapidly they proliferated, and the less collagen and proteoglycan (PG) they produced. The cells that attached after 24 h grew much more slowly, piled up in many areas, exhibited strong alkaline phosphatase activity and calcified extracellular matrix (ECM). Differences in deoxyribonucleic acid (DNA) and protein/PG synthesis were also observed when these cell populations were challenged with growth factors and 12-myristate 13-acetate (PMA). Pretreatment of cells for 2 h with PMA strongly enhanced DNA and PG synthesis only in cultures containing insulin-like growth factor-1. Nonselected rat articular chondrocytes (AC) subcultured at least four times as monolayers still expressed mRNA specific for aggrecan and type II collagen, suggesting conservation of the chondrogenic phenotype. In conclusion, AC of young individuals seem to be heterogeneous with respect to their capacity to proliferate and synthesize ECM. By selecting and expanding in vitro the appropriate cell population, this method could be potentially useful for studies aimed at repairing damaged cartilage.
In vivo Overexpression of Tuftelin in the Enamel Organic Matrix
Authors: Luo Wen , Wen Xin , Wang Hong-Jun , MacDougall Mary , Snead Malcolm L. , Paine Michael L.
Published: October 2004

The primary sequences of human and mouse tuftelin are 89% identical. Both proteins comprise 390 amino acids and produce an acidic protein with an isoelectric point of 5.7, and an unmodified molecular weight of 44 kD. Using fluorescent-tagged tuftelin and amelogenin plasmid constructs we saw little evidence that these two enamel proteins colocalize in ameloblast-like LS-8 cells. Tuftelin is primarily localized to distinct ‘speckled’ domains within the cell cytoplasm. In an attempt to better define a physiological function for tuftelin during amelogenesis, we have produced transgenic mice that overexpress tuftelin in ameloblasts and subsequently the enamel matrix. Tuftelin overexpression impacts dramatically upon the enamel crystallite habit and the enamel prismatic structure. Overexpressing tuftelin results in gross imperfections in enamel that is evident both at the nanoscale and the mesoscale. The most notable difference observed in the transgenic animals, when compared to wild-type animals, is an apparent loss of restricted growth of enamel crystallites along their a-axis and b-axis. This equates to a change in the crystallite aspect ratio. In the transgenic animals the crystallite structures appear more ‘plate’-like in contrast to the symmetric, ‘ribbon’-like crystallite morphology that is a characteristic feature of mammalian enamel.
Expression of Wee1 and Its Related Cell Cycle Components in Mouse Early Stage Follicles
Authors: Park Chang-Eun , Kim Young-Hoon , Jeon Eun-Hyun , Cha Kwang-Yul , Lee Sook-Hwan , Lee Kyung-Ah
Published: October 2004

Wee1 is a kinase regulator of the M-phase promoting factor (a complex of cdc2 and cyclin B1). The present study was performed to determine the role(s) of wee1 in the early stages of mouse ovarian follicles. Expression of wee1 and the correlated cell cycle components, namely cdc2, cyclin B1, and cdc25C, was evaluated by immunohistochemistry. In addition, expression of Tyr15-phosphorylated cdc2 (cdc2-p) was also examined to determine whether wee1 kinase phosphorylates cdc2. Each component except cdc25C was found in the oocyte cytoplasm at all follicular stages, while cdc25C was not detected in primordial follicles. It was found primarily in ovarian interstitial cells and to a small extent in granulosa cells of the developing secondary follicles. To further confirm the expression of cell cycle components in the primordial follicular oocytes, day 1 ovaries were enzymatically and mechanically dissociated, then oocytes were isolated from somatic cells including pre-granulosa cells, and we confirmed that cdc2-p was expressed in oocytes of primordial follicles. The results of the present study led to the conclusion that wee1, without the counteracting cdc25C, would cause meiotic arrest of oocytes by inhibitory phosphorylation of cdc2. Expression of all these proteins in the granulosa cells of growing follicles may regulate granulosa cell mitosis concurrently with the growth of oocytes and follicles.
Expression Patterns of PSA-NCAM in the Human Ganglionic Eminence and Its Vicinity: Role of PSA-NCAM in Neuronal Migration and Axonal Growth?
Authors: Ulfig Norbert , Chan Wood Yee
Published: October 2004

The ganglionic eminence (GE) is a transient but conspicuous structure of the developing forebrain which not only gives rise to a large number of precursor neurons and glial cells for various structures of the forebrain, but in addition serves as an intermediate target for growing axons to the cerebral cortex. To investigate the roles of the highly polysialylated isoform of the neural cell adhesion molecule (PSA-NCAM) in cell migration and axonal growth within the GE and its neighbouring structures, the spatio-temporal expression pattern of PSA-NCAM was examined in human fetal forebrains between 14 and 36 weeks of gestation with a specific immunohistochemical method. Scattered PSA-NCAM-positive cells were found in the centre but more frequently in the marginal zone of the GE. Intensely labelled cells were also identified in the gangliothalamic body, basolateral nuclei of the amygdala and the subventricular and intermediate zone adjacent to the GE. This cellular immunoreactivity started to appear in various structures during the period from 14 to 19 weeks and gradually diminished after 25–28 weeks. Strong immunoreactivity was also detected in fibres running from the intermediate zone of the neocortex to the internal capsule from 16 weeks onwards, and after 24 weeks, the immunoreactivity was gradually decreased. In the vicinity of the GE, between 16 and 22 weeks, short fibre bundles were observed to leave the longitudinally oriented axons of the internal capsule to reach the marginal zone of the GE. Our results suggest a close relationship between PSA-NCAM expression and neuronal migration (over short distances) and transitory axonal projections (target recognition and axonal fasciculation) in the region of the GE.
The Internal Structure of the Nucleus geniculatus lateralis ventralis in the Avian Brain: A Golgi Study and Electron Microscopic Investigation
Authors: Tömböl T. , Eyre M. , Zayats N. , Németh A.
Published: October 2004

Different types of neurons in the ventral geniculate nucleus of the thalamus of chicks were visualised by Golgi impregnation. The dendritic tree of projection neurons branched in a sphere-like territory in both the ventral and middle areas of the lamina externa. The dendrites of projection neurons in the lamina interna descended into the lamina externa and entered both the ventral and middle dendritic areas. One or two dendrites of the lamina interna neurons also emitted branches that developed a dorsal sphere-like dendritic territory. Optic terminals labelled by Golgi impregnation or injection of biotinylated dextran amine were found in these dendritic territories gathered into groups. They established synapses in these areas (synaptic islands or fields without a glial sheath) with different dendritic profiles, and a few gamma-aminobutyric acid (GABA)-positive terminals synapsed with them. No glomerulus-like synaptic complexes ensheathed by glial processes were found. Optic terminals also contacted the stem dendrites of projection neurons and GABA-positive neuron cell bodies and dendrites. Numerous synapses established by both optic and GABA-positive terminals were found on the proximal dendritic stems of the lamina interna projection neurons.
Immunologic Aspects of Chronic Fatigue Syndrome
Authors: Gerrity Timothy R. , Papanicolaou Dimitris A. , Amsterdam Jay D. , Bingham Stephen , Grossman Ashley , Hedrick Terry , Herberman Ronald B. , Krueger Gerhard , Levine Susan , Mohagheghpour Nahid , Moore Rebecca C. , Oleske James , Snell Christopher R.
Published: October 2004

Chronic fatigue syndrome (CFS) is a serious health concern affecting over 800,000 Americans of all ages, races, socioeconomic groups and genders. The etiology and pathophysiology of CFS are unknown, yet studies have suggested an involvement of the immune system. A symposium was organized in October 2001 to explore the possibility of an association between immune dysfunction and CFS, with special emphasis on the interactions between immune dysfunction and other abnormalities noted in the neuroendocrine and autonomic nervous systems of individuals with CFS. This paper represents the consensus of the panel of experts who participated in this meeting. Data suggest that persons with CFS manifest changes in immune responses that fall outside normative ranges, but current research does not provide definitive evidence on whether these immune abnormalities are a cause or result of the illness. It has become clear that CFS cannot be understood based on single measurements of immune, endocrine, cardiovascular, or autonomic nervous system dysfunction. This panel encourages a new emphasis on multidisciplinary research into CFS.
Vasoactive Intestinal Peptide Can Modulate Immune and Endocrine Responses during Lipopolysaccharide-Induced Acute Inflammation
Authors: Bik Wojciech , Wolinska-Witort Ewa , Chmielowska Magdalena , Baranowska-Bik Agnieszka , Rusiecka-Kuczalek Elzbieta , Baranowska Boguslawa
Published: October 2004

Objectives: In many studies, it has been reported that vasoactive intestinal peptide (VIP) may play an important role in modulation of the immunological response. VIP can be produced by immunological cells, and also the receptors for this neuropeptide are present in many of these cells. The aim of our study was to estimate the effects of the administration of exogenous VIP on serum concentrations of proinflammatory cytokines [interleukin (IL)-6 and tumor necrosis factor (TNF)-α] and an anti-inflammatory cytokine (IL-10) during lipopolysaccharide (LPS)-induced acute inflammation. We also estimated the influence of VIP on pituitary [FSH, LH, TSH and prolactin (PRL)], thyroid (T3 and T4), adrenal (corticosterone) and gonadal (testosterone) hormones in response to LPS-induced acute inflammation. Methods: Male Wistar-Kyoto rats were divided into four groups, which received, respectively, placebo (0.9% NaCl), LPS, VIP and VIP with LPS. The TNF-α and IL-6 serum concentrations were measured after 2 h from the time of the administration of the agents, IL-10 was measured after 4 h, and the pituitary, thyroid, adrenal and gonadal hormone concentrations were measured after 2 and 4 h. Cytokine concentrations were estimated using ELISA tests, and hormone concentrations were measured using RIA tests. Results: In our experiments, LPS administration dramatically increased serum proinflammatory cytokine concentrations (TNF-α and IL-6) after 2 h and the anti-inflammatory cytokine (IL-10) after 4 h, as well as increasing the serum corticosterone concentration (after 2 and 4 h) and LH (after 2 h). LPS application decreased serum concentrations of T3 and TSH (both after 2 h), testosterone (after 2 and 4 h), FSH after 4 h and PRL after 4 h. VIP administration decreased the serum IL-10 concentration after 4 h and T3 concentration after 2 h and increased serum concentrations of FSH and corticosterone after 4 h. VIP administrated simultaneously with LPS decreased the LPS-induced increase in IL-6 and corticosterone concentrations (consecutively after 2 and 4 h). VIP also enhanced LPS-induced thyroid hormone (T3 and T4) suppression after 4 h and testosterone suppression after 4 h. Conclusion: We conclude that VIP can modulate not only immune responses but also hormonal responses during acute inflammation.
The Influence of Immunomodulation on Psycho-Neuroimmunological Functions in Benign Multiple Sclerosis
Authors: Haase Claus G. , Tinnefeld Michael , Faustmann Pedro M.
Published: October 2004

Objectives: In multiple sclerosis (MS), several neuroimmunomodulatory effectors are known, including melatonin. They are able to influence disease-related neurophysiogical changes (disability or impaired vision) as well as neuropsychological performance (e.g. cognition and depression). In this study we assessed the relationship between immunomodulation on psycho-neuroimmunological functions in benign multiple sclerosis. Methods: We evaluated 26 young female patients with benign MS treated with/without immunomodulating therapies with regard to their physical disabilities (Expanded Disability Status Scale, EDSS), their visually evoked potentials (VEP), their plasma melatonin concentrations as well as their performance regarding emotional and cognitive tests and compared them with healthy matched controls. Results: Patients with MS showed deficits in cognitive and emotional functions compared to healthy controls, which were in accordance with their increase in EDSS over time. However, in contrast to untreated patients, patients receiving immunotherapy showed significantly increased dysfunction with respect to actual mood (p = 0.02) and a tendency to increased depression scores (p = 0.072). However, neither treatment subgroup had cognitive deficits. In untreated patients, melatonin levels correlated with reduced scores in the cognitive tests (p = 0.045) but not with depression or VEP latencies. Patients with long-standing MS (>10 years) showed a significant correlation (p = 0.01) to their increased depression scores and their melatonin levels, but no correlation with VEP or cognitive dysfunction, compared to patients with shorter disease duration (≤10 years). Conclusion: These results indicate that in MS all aspects of the psycho-neuroimmunological network can be affected. Despite the potential influence of immunomodulation on depression, no connection with melatonin representing the retinohypothalamic tract/pineal gland circuits could be detected. However, visual perception as well as visuoconstructive abilities were affected in MS patients. Neuropsychological tests in MS should concentrate on cognitive variables, which reflect the clinical status more accurately and may be used to monitor disease-modifying therapies.
Inhalation of Low-Level Formaldehyde Enhances Nerve Growth Factor Production in the Hippocampus of Mice
Authors: Fujimaki Hidekazu , Kurokawa Yoshika , Kakeyama Masaki , Kunugita Naoki , Fueta Yukiko , Fukuda Takaichi , Hori Hajime , Arashidani Keiichi
Published: October 2004

Objective: The effects of low-level formaldehyde (FA) inhalation on the amount of nerve growth factor (NGF) in the hippocampus of immunized mice were studied. Methods: Evaluation of NGF in the hippocampus was performed by ELISA and RT-PCR. Results: Exposure to 80 and 400 ppb FA significantly increased the brain NGF levels in the immunized mice. Evaluation of the NGF levels in the hippocampus of immunized mice showed that 400 ppb FA significantly increased the NGF content. The RT-PCR evaluation also showed higher concentrations of hippocampal NGF mRNA in the mice exposed to 80 and 400 ppb FA with immunization. Conclusion: Exposure of immunized mice to low levels of FA significantly increases NGF levels in the hippocampus.
IL-1α and TNF-α Expression in Rat Periapical Lesions and Dental Pulp after Unilateral Sympathectomy
Authors: Bletsa Athanasia , Heyeraas Karin Johanne , Haug Sivakami Rethnam , Berggreen Ellen
Published: October 2004

Objectives: Apical periodontitis is an inflammatory disease characterized by bone resorption, and sympathetic nerves are known to modulate bone resorption and bone remodeling. Higher numbers of osteoclasts and larger periapical lesions have been observed after sympathectomy in rats, but the mechanisms underlying the inhibitory effect of sympathetic nerves on osteoclasts are unknown. This study aimed to test the hypothesis that sympathetic nerves inhibit the production of the bone-resorbing pro-inflammatory cytokines IL-1α and TNF-α in rat periapical lesions. Methods: Rats were unilaterally sympathectomized and apical lesions were induced by exposing the dental pulp of molar teeth to the oral microflora. We quantified the cytokines IL-1α and TNF-α by enzyme-linked immunosorbent assay, and immunohistochemical analysis was done for qualitative localization. Pulp from intact incisor teeth was tested as a control. Results: We showed that IL-1α was increased, but not TNF-α, in the periapical lesions on the sympathectomized side. Both IL-1α and TNF-α were expressed in unexposed pulp. TNF-α was significantly decreased in the denervated incisor pulp, whereas the level of IL-1α remained unchanged. Conclusions: This study suggests that sympathetic nerves have an inhibitory effect on IL-1α in periapical lesions and a stimulatory effect on TNF-α in the intact rat pulp.
The Role of Asymmetry of Nervous and Immune Systems in the Formation of Cellular Immunity of (CBaxC57Bl/6) F1 Mice
Authors: Gontova Irina A. , Abramov Valery V. , Kozlov Vladimir A.
Published: October 2004

We have previously shown the existence of functional asymmetry of the immune system and the role of brain hemispheres and different lobes of thymus in the development of humoral immune response in (CBA×C57Bl/6) F1 mice. The role of asymmetry of the nervous and immune systems in the formation of the cellular immune response [delayed-type hypersensitivity (DTH) reaction] in these mice has been studied in our work. In order to test the influence of asymmetry of the primary immune organs, thymus, on the cellular immune response, mice were thymectomized and then we studied the effect of the injection of cells from contralateral thymus lobes of right-dominant and left-dominant donors by motor asymmetry on how pronounced the DTH reaction in the back left paw was. The injection of thymocytes from right-dominant donors appeared to result in significant differences in DTH reaction between left- and right-handed mice. At the same time, our experiments failed to discover any pronounced role of thymus asymmetry in the formation of DTH reaction. In order to test the influence of asymmetry of peripheral immune organs, regional lymph nodes, on the regulation of cellular immune response, we compared the DTH reaction in left and right paws of mice. We found that the intensity of the DTH reaction to sheep red blood cells in the front paws of (CBA×C57Bl/6) F1 mice depends not only on whether the antigen is injected into the left or right paw but also on the motor asymmetry of the hemispheres. While comparing the DTH reaction in the back left and right paw of mice we showed that in both right- and left-handed mice it was much more pronounced in the left paw than in the right one. The data obtained testify to the functional asymmetry of bilateral lymph nodes located near the forming cellular immune reaction. Thus, the results obtained show that the intensity of DTH reaction in (CBA×C57Bl/6) F1 mice depends on the functional asymmetry of regional lymph nodes and motor of brain hemispheres. The thymus functional asymmetry is of slight importance in DTH reaction.
Methionine-Enkephalin Stimulates Hydrogen Peroxide and Nitric Oxide Production in Rat Peritoneal Macrophages: Interaction of μ, δ and κ Opioid Receptors
Authors: Vujić Vesna , Stanojević Stanislava , Dimitrijević Mirjana
Published: October 2004

Objective: Methionine-enkephalin (MET) modulates various functions of macrophages related to both immune and inflammatory reactions in a naloxone reversible manner, suggesting that opioid receptors are involved in the regulation of macrophage activity. Since an endogenous opioid ligand might interact with more than one type of opioid receptor, the receptor interaction determines its effect on a particular function. Methods: In the present study we have investigated the involvement of different opioid receptor types/subtypes in MET-induced modulation of H2O2 and NO production in macrophages. Thioglycollate-elicited or resident rat peritoneal macrophages were treated in vitro with MET and/or specific antagonists of δ1,2, δ1, δ2, µ and ĸ opioid receptors. Results: MET increased H2O2 production inphorbol myristate acetate-stimulated rat peritoneal macrophages mainly through δ1 opioid receptor. MET also enhanced NOproduction in rat peritoneal macrophages stimulated with lipopolysaccharide through δ1 and µ opioid receptors. The blockade of µ and ĸ receptor facilitated a potentiating effect of MET on H2O2 release, and blockade of ĸ receptor further raised the MET-induced increase of NO production in macrophages. Conclusion: It is concluded that both negative and positive functional interaction between δ, µ and ĸ opioid receptors regulate the influence of MET on H2O2 and NO production in rat peritoneal macrophages.
Dexamethasone Downregulates Chemokine Receptor CXCR4 and Exerts Neuroprotection against Hypoxia/Ischemia-Induced Brain Injury in Neonatal Rats
Authors: Felszeghy Klára , Banisadr Ghazal , Rostène William , Nyakas Csaba , Haour France
Published: October 2004

Objective: Hypoxia/ischemia (H/I) induces rapid and massive brain damage in neonatal rat brain, resulting in long-term consequences on structural and functional maturation of the central nervous system. Inflammatory mediators contribute to these permanent pathological changes, which are sensitive to corticoid treatments. Since the chemokine receptor CXCR4, specific for the SDF-1α/CXCL12 ligand, regulates both apoptotic and neuroregeneration processes, this receptor was quantified 2 days following H/I in neonatal rat brain in relation with dexamethasone (DEX) treatment. Methods: Seven-day-old male rats were exposed to a 90-min hypoxia following unilateral carotid ligation (H/I) and were sacrificed 48 h later. Glucocorticoid-pretreated animals were injected subcutaneously 5 h prior to hypoxia with 0.5 µg/g DEX. Glial fibrillary acidic protein and cresyl violet staining were used for assessing the extent of brain lesion subdivided into necrotic and penumbra-like areas. The density of CXCR4 receptors was determined by quantitative autoradiography using [125I]SDF-1α as a ligand. Results: The H/I resulted in a massive lesion ipsilateral to the carotid ligation, which was extended to cortical, striatal, hippocampal and thalamic areas, while the contralateral hemisphere remained apparently unaffected. DEX decreased the lesion size by reducing mainly the necrotic area. H/I induced a marked increase in CXCR4 receptor binding in the penumbra-like areas. DEX pretreatment decreased CXCR4 receptor density in the penumbra and attenuated astrocytosis. Furthermore, DEX strongly lowered mortality rate and reduced functional recovery time right after hypoxia. Conclusion: The rapid enhancement in CXCR4 chemokine receptor binding in the affected brain areas suggests that SDF-1α/CXCR4 may play a role in the hypoxia-induced inflammatory reaction in the neonatal brain. Attenuation of CXCR4 expression and astrogliosis could contribute to the neuroprotective effect of DEX pretreatment via influencing the inflammatory cascade induced by H/I in the neonatal brain.
Immune Alterations Induced by Social Defeat Do Not Alter the Course of an On-Going BCG Infection in Mice
Authors: Merlot Elodie , Moze Elisabeth , Dantzer Robert , Neveu Pierre J.
Published: October 2004

The timing for applying stressor and primary immunization is known to influence the nature of the immune alterations induced by stress. The aim of the present study was to investigate the consequences of a stress occurring several days after the beginning of a primary infection on the host resistance. For this purpose, we investigated the effects of repeated social defeat on the immune response of mice infected with BCG 11 days before. In vitro production of cytokines in response to LPS or tuberculin, and the sensitivity of spleen cells to corticosterone were assessed 8 days after the end of the stress. Bacterial growth was assessed in the spleen. We demonstrated that social defeat in BCG-infected mice induced a long-term increase in IL-6 and IL-10 production in response to LPS but did not modify the sensitivity of spleen cells to corticosterone. Stress did not affect the specific response to BCG, as shown by the production of cytokines in tuberculin-stimulated cultures. Accordingly, social defeat was unable to influence the mycobacterial growth in vivo. These results support the hypothesis postulating that stress does not affect antigen-specific response when it is applied after priming.
Electric Foot-Shock Stress Drives TNF-α Production in the Liver of IL-6-Deficient Mice
Authors: Chida Yoichi , Sudo Nobuyuki , Motomura Yoshitomo , Kubo Chiharu
Published: October 2004

Objectives: Accumulating evidence has shown that interleukin-6 (IL-6) has pleiotropic effects on a variety of biological functions, including its antiapoptotic potential during liver injury. Our previous work demonstrated that restraint stress-induced elevation of plasma IL-6 negatively regulates plasma tumor necrosis factor-α (TNF-α). Herein, we further clarified the mechanism underlying the above finding and investigated the effect of IL-6 on liver apoptosis triggered by stress. Methods: Male C57BL/6J and IL-6-deficient C57BL/SV129 mice were exposed to 1 h of electric foot-shock stress. Thereafter, the serum, liver and spleen TNF-α levels were measured at several time points. Serum alanine aminotransferase (ALT), liver caspase-3 and terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling (TUNEL) activities were analyzed to evaluate the severity of liver injury and apoptosis. Results: The liver, but not the spleen, of the IL-6-deficient mice exhibited a significant increase in TNF-α level after stress in parallel with serum TNF-α elevation, whereas no such TNF-α responses were found in the wild animals. No significant differences in stress-induced elevation of serum ALT levels, liver caspase-3 activities and the number of TUNEL-positive hepatocytes were found between the wild and IL-6-deficient mice. Conclusions: Taken together, these results indicate that IL-6 may play a critical role in suppressing TNF-α production in the liver, thereby decreasing the blood TNF-α level. In contrast, IL-6 secretion was shown to have no protective effect on stress-triggered liver injury.
c-Fos Expression in the Rat Cerebral Cortex during Systemic GvH Reaction
Authors: Furukawa Hiromu , Yamashita Akira , del Rey Adriana , Besedovsky Hugo
Published: October 2004

Objective: It is becoming clear that the CNS receives signals from the peripheral immune system. In order to identify the areas of the brain that receive information about a specific immune response to allogeneic antigens, we studied the expression of c-Fos, a neural activation marker, in the cerebral cortex following the induction of a graft-vs.-host reaction (GvHR) in rats. Methods: C-Fos expression in the brain was studied by immunohistochemistry. GvHR was induced in (WKY × PVG)F1 rats by injecting 5 × 108 spleen cells from PVG rats. Control rats received syngeneic cells. Results: No c-Fos immunoreactivity (IR) was observed in animals undergoing GvHR in the nucleus tractus solitarii (NTS), the locus coeruleus (LC), the organum vasculosum of lamina terminalis (OVLT), the paraventricular nucleus (PVN) or the central amygdaloid nucleus (Ce). In contrast, 3 days after GvH induction c-Fos IR was observed in the piriform cortex and several other olfactory-related regions indicating the stimulation of the olfactory pathway during GvHR. Strong c-Fos IR was also observed in the occipital visual cortex of animals undergoing a GvHR, suggesting that GvHR can affect visual functions. In addition, GvHR induced c-Fos IR in the prefrontal cortex (Cg3, orbital cortex), a region that has interconnections with most sensory modalities. Double-staining studies indicate that the cells that express the c-Fos signal are neurons. Conclusion: We have defined the distribution of brain neurons that are affected during the induction phase of GvHR. Our results also indicate that the integration and processing of information from the immune system at CNS levels involve different areas during different types of immune responses.
Screening, Diagnosis and Advances in Aortic Aneurysm Surgery
Authors: Daly Kevin J. , Torella Francesco , Ashleigh Ray , McCollum Charles N.
Published: October 2004

Background: Aortic aneurysms are common in the elderly and a frequent cause of sudden death. As elective aneurysm repair has a mortality drastically lower than that associated with rupture, the emphasis must be on early detection and repair free from complications. Recent advances include ultrasound screening for asymptomatic abdominal aortic aneurysm (AAA) and clinical trials on the size of AAA that require repair. Pre-operative assessment, management of cardiac risk, autologous blood transfusion strategies, and endovascular stent graft technology to avoid major open surgery are all issues to be addressed. Methods: Following a computerized Medline search for publications on the detection and treatment of abdominal and thoracic aortic aneurysm, the publications identified were then read and the references within those publications examined for further publications on this topic. We have reviewed these publications without attempting a meta-analysis. Results: Randomized population studies have addressed ultrasound screening for AAA. Attendance for screening was good and AAA detection inexpensive. Screening men from 65 years reduces the mortality from rupture and is cost-effective. Open thoracic and abdominal aneurysm repair has a mortality of around 8%, with myocardial infarction being a frequent cause of death. Pre-operative reduction of cardiac risk by cardiac investigations and β-blockade may reduce this mortality. Autologous transfusion techniques such as acute normovolaemic haemodilution and interoperative cell salvage reduce the need for allogeneic blood and the complications associated with open surgery. Minimally invasive endovascular repair is now possible for 40% of the AAA and an increasing proportion of thoracic aneurysms. Conclusions: The combination of screening, reduced pre-operative risk, and new minimally invasive techniques extends aortic aneurysm treatment into an increasingly elderly population. The combination of these techniques will reduce mortality from ruptured aortic aneurysm in the elderly and also reduce the stress associated with aneurysm surgery.
Aging Changes of the Angioarchitecture and Arterial Morphology of the Spinal Cord in Rats
Authors: Qiu Ming-Guo , Zhu Xing-Hong
Published: October 2004

Background: It is well known that vascular myelopathy increases with age, however, there are no systematic reports on the quantitative capillary density in the spinal cord and no reports concerning the age-related changes of vascularity and capillary density of the spinal cord. Objective: Age-related changes of vascularity and capillary density of the spinal cord and the morphology of the ventral spinal artery were observed and measured so as to provide reference to study the microcirculation of the spinal cord. Methods: Three different age groups of Wistar rats (1–2, 6–7 and >22 months old) were studied by ink-gelatin injection, corrosion casts, biological stereological technique, image analysis and transmission electron microscope. Results: The number of central arteries per centimeter of the spinal cord decreased significantly with age; the ventral spinal artery and central arteries often became tortuous in the >22 month group. Capillary density in the anterior horn was significantly higher than that in the posterior horn. Capillary density in the gray matter decreased significantly with age. These aging changes of the ventral spinal artery in the rat included that the folds of internal elastic membrane were reduced, the internal elastic membrane thinned and disrupted, the endothelial cells degenerated and broke away from the wall, and smooth muscle cells in the tunica media degenerated and protruded to the lumen. The relative content of the elastic fibers decreased significantly with age, while the relative content of the collagenic fibers and the ratio of the C/E increased significantly with age. Conclusions: The difference between capillary density in gray and white matter in the spinal cord has shown that the need of metabolization is adapted to the function of the neurons, and the anterior horn is more sensitive than the posterior horn to anoxia and is easily damaged during ischemia in the spinal cord. Aging changes of capillary density and arterial morphology of the spinal cord may be important contributory factors in vascular myelopathy.