bluuebunny/crossref_metadata_2025 huggingface dataset:
Plasma Homocyst(e)ine Concentrations and the Risk of Subtypes of Cerebral Infarction
Authors: Shimizu Haruki , Kiyohara Yutaka , Kato Isao , Tanizaki Yumihiro , Ueno Hirofumi , Kimura Yasuhiro , Iwamoto Hiromitsu , Kubo Michiaki , Arima Hisatomi , Ibayashi Setsuro , Fujishima Masatoshi
Published: April 2003

Moderately elevated plasma total homocyst(e)ine (tHcy) levels have been linked with cardiovascular disease. However, the findings of previous studies regarding the relationship between tHcy levels and subtypes of cerebral infarction (CI) have been conflicting. The aim of the present study was to examine this issue in a community-based case-control study performed in Hisayama Town in Japan. Fasting tHcy levels were compared among 75 CI cases, of which 43 were lacunar (LI), 24 atherothrombotic (ATI) and 8 cardioembolic infarctions (CEI), and 248 age- and sex-matched healthy controls. The mean tHcy concentrations were higher in CI than in controls (13.0 vs. 11.8 µmol/l; p = 0.018). LI and CEI also had significantly higher tHcy levels than did the corresponding controls (12.3 vs. 11.3 µmol/l for LI; p = 0.037 and 16.3 vs. 12.7 µmol/l for CEI; p = 0.036). The same tendency was also observed for ATI, but the difference was only marginally significant probably due to the small number of the cases (13.4 vs. 11.9 µmol/l; p = 0.087). After adjustment for age, sex, hypertension, serum creatinine, total protein, folate and vitamin B12 levels, the risk of LI was not significant in the second tertile of the tHcy distribution, but significantly increased in the third compared with the first tertile (adjusted odds ratio, AOR, 3.4; 95% confidence limits, CL, 1.3–8.9; p = 0.015), while the risk of ATI was significant even in the second tertile (AOR, 5.0; 95% CL, 1.0–23.7; p = 0.042) and higher in the third tertile (AOR, 7.5; 95% CL, 1.5–38.3; p = 0.015). However, the odds ratios for CEI could not be estimated, as there was no case of CEI in the first tertile. These findings suggest that elevated tHcy is an independent risk factor for all subtypes of CI, but that its impact is higher in ATI and probably in CEI than in LI.
Sonographic Characterisation of Radiation-Induced Carotid Artery Stenosis
Authors: Lam W.W.M. , Liu K.H. , Leung S.F. , Wong K.S. , So N.M.C. , Yuen H.Y. , Metreweli C.
Published: April 2003

Background and Purpose: To study the distribution, extent and sonographic characterisation of radiation-induced carotid artery stenosis in nasopharyngeal carcinoma (NPC) patients. Methods: The distribution of plaques, the extent of stenosis, and the sonographic characterisation of the plaque at maximum stenosis were recorded in 71 NPC patients. The results were compared with the ultrasound results of a control group of 142 patients presenting with symptoms of cerebrovascular disease or carotid bruit. Results: NPC patients had a higher incidence of carotid stenosis (77 vs. 50.7%). The common carotid arteries were most commonly affected by radiation-induced stenosis (93/142 vs. 37/284 in the control group), whereas the carotid bulb was the most commonly affected (56/284) site in the control group. Significantly more NPC patients had moderate-to-severe stenosis (21/71 vs. 27/142). Analysis of the sonographic appearance of radiation-induced and atherosclerotic plaques showed more diffuse involvement in the post-radiation group. Non-calcified plaques and intraplaque hypoechoic foci were also more frequent in the post-radiation group. Conclusions: Radiation-induced carotid stenosis is more diffuse in distribution, is associated with more severe luminal stenosis and has different sonographic plaque characterisation compared with carotid stenosis without radiation exposure.
Evaluation of Progression and Spread of Atherothrombosis
Authors: Droste Dirk W. , Ringelstein E. Bernd
Published: April 2003

Symptomatic atherothrombosis in one vascular bed is usually indicative of disseminated disease. Indeed, involvement of multiple beds is common in everyday clinical practice, and these patients are at much higher risk of ischaemic events. The prevention of manifestations following atherothrombosis is therefore an important therapeutic goal in these patients. Some causative risk factors demonstrate affinities to particular arterial domains. Cigarette smoking, for example, is particularly associated with atherothrombotic involvement of the pelvic and lower limb arteries, whereas arterial hypertension is associated with the intracranial cerebral arteries. The degree, spread and progression of atherosclerosis can be assessed using various non-invasive and invasive modalities: high-resolution Doppler ultrasound, ankle-brachial index (ABI) measurement, magnetic resonance (MR), computed tomography (CT) and intra-arterial angiography. Indicators of atherothrombotic risk include increased carotid artery intima-media thickness, microembolic signals on transcranial Doppler ultrasonography and low ABI. There is a strong rationale for the inclusion of the ABI measurement as part of the routine clinical examination to assess the cardiovascular risk in patients with identified risk factors. Furthermore, detection of a low ABI should serve as a trigger for patient management with aggressive antiplatelet therapy. The generalized nature of atherothrombosis and the methods for evaluating the spread of disease are illustrated through the case history of a patient with disseminated atherothrombotic disease.
Morphology of Isolated Colonic Crypts
Authors: Robert Marie E. , Singh Satish K. , Ikuma Mitsuhiro , Jain Dhanpat , Ardito Thomas , Binder Henry J.
Published: April 2003

The traditional paradigm of colonic fluid and electrolyte transport includes a spatial separation of absorptive and secretory processes to surface and crypt cells, respectively. Recent studies of isolated microperfused colonic crypts revealed constitutive Na-dependent fluid absorption while secretion is regulated by one or more neurohumoral agonists. One obvious reason for the difference found in microdissected crypts is their separation from the lamina propria milieu. While it has been shown that isolated crypts are devoid of obvious lamina propria elements, including pericryptal fibroblasts, detailed morphologic information of the content of isolated crypts has been lacking. To characterize the morphology of the isolated crypt, we performed transmission electron microscopy (TEM) and immunofluorescence on microdissected and Ca2+ chelated crypts. Crypt cell type analysis was carried out separately on intact rat colon using light microscopy. TEM revealed a complete lack of either lamina propria cells or extracellular material in crypts isolated by either technique. TEM also revealed a subtle difference between the two isolation methods, with intact basal membranes in microdissected crypts but focal disruption of basal membranes in Ca2+- chelated crypts. Immunofluorescent stains for two basement membrane components (laminin and collagen type IV) revealed the presence of adherent basement membrane only on microdissected crypts; evidence that the plane of separation differs in these two preparations. Crypt cell type analysis on intact rat colon revealed an equal proportion of goblet cells in the right and left colon (approximately 50%) when measuring the middle 70% of the crypts – the area studied during crypt microperfusion. This morphologic analysis will increase our understanding of the observed physiology of isolated colonic crypts.
Sex Differences in Brainstem Neural Activation after Injury to the TMJ Region
Authors: Bereiter David A.
Published: April 2003

The basis for a higher prevalence of painful temporomandibular disorders (TMD) among women than men is not known. The present study used Fos-like immunoreactivity (Fos-LI) to quantify the pattern and magnitude of neural activation within the trigeminal brainstem complex of male and female rats caused by acute inflammatory injury to the temporomandibular joint (TMJ) region. Also, Fos-LI was assessed in animals given morphine, a preferential mu opioid receptor agonist, or U50,488H, a selective kappa opioid agonist, prior to TMJ injury to determine if opioid modulation of neural activation was similar in males and females. The general pattern of Fos-LI after TMJ injury was similar in males and females. This pattern was characterized by a high density of Fos-positive neurons in the dorsal paratrigeminal nucleus (dPa5), subnucleus interpolaris/caudalis transition region (Vi/Vc-vl), and in the superficial laminae at the subnucleus caudalis/upper cervical spinal cord (Vc/C2) junction ipsilateral to TMJ injury. In contrast to other regions the number of Fos-positive neurons produced at the Vc/C2 junction was proportional to the concentration of mustard oil injected into the TMJ region. In addition, proestrus females produced higher levels of Fos-LI at the Vc/C2 junction than diestrus females or males. Morphine caused a greater dose-related reduction in Fos-LI at the dPa5 and Vc/C2 junction in males than females. By contrast, U50,488H caused a dose-related reduction in Fos-LI only at the Vc/C2 junction of proestrus females. These results support the hypothesis that the Vc/C2 junction region plays a critical role in the integration of pain signals originating from the TMJ region and may underlie sex differences in sensory processing related to TMJ pain.
Drug Discovery and Target Validation
Authors: Nuttall Mark E.
Published: April 2003

Recent drug discovery has been driven largely by a genomics-based approach. This revolution in pharmaceutics is based on localized expression of either a novel gene or homologue of a known gene found in cDNA libraries made from normal versus diseased tissue. The choice and quality of cDNA library is critical for the success of this approach. Expression is normally verified at the cellular level by either immunocytochemistry or in situ hybridization. Activity of the recombinant protein in secondary cell-based assays allows highthroughput screens to be formulated to identify small-molecule effectors of this protein. More recently, a proteomics approach has also been incorporated into this process. This technology directly measures proteins whose expression is localized in disease tissue as the basis for cell-based screens to look for either activators or inhibitors, of this activity. The majority of screens are designed to look for inhibitors. Activity of small-molecules found by screening gives rise to pharmacokinetic studies and verification of activity in animal models of the disease. Structure-activity relationship (SAR) optimization of these small-molecules allows for suitable oral bioavailability and pharmacokinetics, resulting in compounds progressing from discovery to development. Based on these strategies, we have developed inhibitors of osteoclast-mediated bone resorption and are currently screening for bone anabolic agents. In addition, we have also developed small-molecule caspase inhibitors which prevent chondrocyte apoptosis and retain cell function in an attempt to find therapeutic agents to either prevent or treat osteoarthritis. These agents may well have utility in the treatment of temporomandibular joint diseases.
Muscle Spindles in Extraocular Muscles of Human Infants
Authors: Bruenech J. Richard , Ruskell Gordon L.
Published: April 2003

The capacity of muscle spindles in adult human extraocular muscles (EOM) to provide effective proprioception was questioned on the grounds of their peculiar morphology. Their appearance could be attributable to ageing and to test this possibility examples of infant muscle spindles have been examined. Forty encapsulated structures from five extraocular muscles removed post mortem from 4 infant patients aged 6 days, 5, 23 and 30 months were examined by means of light microscopy using serial transverse sections. Seven of them were identified as false spindles. The remaining 33 structures, identified as spindles, contained a total of 175 intrafusal fibres varying from 2 to 12 (mean: 6) in each. 130 of these fibres (74.3%) were of nuclear chain type. Unequivocal evidence of bag fibres was not found. Spindles lacked or had a limited equatorial expansion, and the inner capsule was incomplete and irregularly shaped. 45 (25.7%) of the intrafusal fibres had extrafusal features with large diameters, peripherally placed nuclei, no equatorial modification and without associated sensory nerve terminals. Serial sections revealed that a majority of the nuclear chain fibres were interrupted, fragmented or terminated abruptly, and most spindles contained at least one incomplete fibre. These observations show that the atypical features observed in adult human EOM spindles are also present in infants and are therefore not attributable to ageing.
Electron Microscopy of the Canine Corneal Basement Membranes
Authors: Abrams George A. , Bentley Ellison , Nealey Paul F. , Murphy Christopher J.
Published: April 2003

The purpose of this study was to characterize the surface topographical features of the epithelial and endothelial (Descemet’s) basement membranes of the canine cornea. Corneas were obtained from young, healthy dogs (<2 years old) with no history or evidence of previous ocular disease. The epithelium and endothelium was carefully removed preserving the anterior and posterior basement membranes. The specimens were examined by transmission electron microscopy and scanning electron microscopy. The epithelial and endothelial basement membrane surface topography is an intricate meshwork of pores and fibers measuring in the nanometer size range. The features of the endothelial basement membrane overall are smaller in size than the epithelial basement membrane. These surface topographical features may incite changes in epithelial and endothelial cell behavior.
Study of the Association between Alzheimer’s Disease and Angiotensin-Converting Enzyme Gene Polymorphism Using DNA from Lymphocytes
Authors: Cheng Chih-Ya , Hong Chen-Jee , Liu Hsiu-Chih , Liu Tsung-Yun , Tsai Shih-Jen
Published: April 2003

Angiotensin-converting enzyme (ACE) may play a role in cognition and memory. A recent study found that a 287-bp insertion/deletion (I/D) polymorphism of the ACE gene is associated with susceptibility to Alzheimer’s disease (AD). However, this finding has not been replicated by all studies. These discrepancies may be due to the difference in ethnic background. Therefore, we investigated the possibility of such an association in 173 AD patients and 286 normal controls from a Chinese population. We found that there is an increased frequency of the ACE I allele in AD patients and the odds ratio for bearers of the I allele was 2.88. Our findings further support previous reports of an association between ACE polymorphism and AD. The implication of the ACE I/D polymorphism in the pathogenesis of AD warrants further exploration.
Integrated Skin Protection from Workplace Irritants: A New Model for Efficacy Assessment
Authors: Berndt U. , Gabard B. , Schliemann-Willers S. , Wigger-Alberti W. , Zitterbart D. , Elsner P.
Published: April 2003

Background: Up to date, the benefit of the combined use of different work-process-depending protective measures for the prevention of occupational irritant contact dermatitis has not yet been proven scientifically. Objective: The study was aimed at assessing the efficacy of an integrated skin protection from workplace irritants. Methods: A repetitive irritation test with sodium lauryl sulphate was used to induce an irritant contact dermatitis on the forearms of 40 subjects. Preventive measures, i.e. barrier cream, skin cleanser and skin care cream, were applied alone and together in different combinations. The resulting protective effects were evaluated clinically and by bio-engineering methods. Results and Conclusion: (1) The highest benefit was achieved if all three protective measures were combined. (2) Thus, the efficacy of the integrated skin protection was confirmed. (3) However, the use of barrier cream appeared to be the most important part.
Vampire Bat Plasminogen Activator DSPA-Alpha-1 (Desmoteplase): A Thrombolytic Drug Optimized by Natural Selection
Authors: Schleuning Wolf-Dieter
Published: April 2003

Plasminogen activators are enzymes found in all vertebrate species investigated so far. Their physiological function is the generation of localized proteolysis in the context of tissue remodeling, wound healing and neuronal plasticity. The common vampire bat (Desmodus rotundus) is a New World species that feeds exclusively on blood. Its saliva contains highly potent plasminogen activators, specialized in rapid lysis of fresh blood clots. Biochemical and pharmacological evidence indicates that these plasminogen activators represent a new class of thrombolytics with pharmacological and toxicological properties superior to human tissue-type plasminogen activator, the clot dissolving agent now most frequently used in medicine. A form of the enzyme produced by recombinant DNA technology is currently employed to test this hypothesis in clinical studies.
A Novel Snake Venom Disintegrin That Inhibits Human Ovarian Cancer Dissemination and Angiogenesis in an Orthotopic Nude Mouse Model
Authors: Markland Francis S. , Shieh Kate , Zhou Qing , Golubkov Vladislav , Sherwin Russell P. , Richters Valda , Sposto Richard
Published: April 2003

OVCAR-5 is a human epithelial carcinoma cell line of the ovary, established from the ascitic fluid of a patient with progressive ovarian adenocarcinoma without prior cytotoxic treatment. The unique growth pattern of ovarian carcinoma makes it an ideal model for examining the anticancer activity of contortrostatin (CN), a homodimeric disintegrin from southern copperhead venom. FACS analysis revealed that OVCAR-5 is integrin αvβ3 negative, but αvβ5 positive. CN effectively blocks the adhesion of OVCAR-5 cells to several extracellular matrix proteins and inhibits tumor cell invasion through an artificial basement membrane. In a xenograft nude mouse model with intraperitoneal introduction of OVCAR-5 cells, intraperitoneal injection of CN was used for therapy. Tumor dissemination in CN-treated versus control groups was studied by gross examination, and antiangiogenic potential was examined by factor VIII immunohistochemistry and image analysis. CN not only significantly inhibited ovarian cancer dissemination in the nude mouse model, but it also dramatically prevented the recruitment of blood vessels to tumors at secondary sites.
Interaction of Bothrojaracin with Prothrombin
Authors: Zingali Russolina B. , Bianconi M. Lucia , Monteiro Robson Q.
Published: April 2003

Bothrojaracin (BJC) is a 27-kD protein from Bothrops jararaca venom that interacts with α-thrombin (KD = 0.7 nM) through both anion-binding exosites I and II. Recently, it has been shown that BJC interacts with the exosite I precursor (proexosite I) on human prothrombin (KD = 75 nM), forming a 1:1 Ca2+-independent noncovalent complex with the zymogen. Complex formation was associated with inhibition of zymogen activation by Oxyuranusscutellatus venom. In addition, BJC strongly decreased the prothrombin activation by factor Xa only in the presence of factor Va. A similar effect was observed in the presence of phospholipids, suggesting that BJC specifically inhibits the interaction of prothrombin with factor Va. It is proposed that BJC has two independent mechanisms for anticoagulation: (1) inhibition of exosite-I-dependent activities on α-thrombin, and (2) inhibition of prothrombin activation through interaction with proexosite I.
Lonomia Genus Caterpillar Envenomation: Clinical and Biological Aspects
Authors: Arocha-Piñango Carmen Luisa , Guerrero Belsy
Published: April 2003

Persons who have been in contact with Lonomia achelous or Lonomia obliqua caterpillars present external and internal bleeding and opening of recently healed wounds. Hematological tests show normal platelet count, prolonged prothrombin time, activated partial thromboplastin time and thrombin time, totally corrected by normal plasma. Decreased fibrinogen (Fg), factor (F) V, FXIII, plasminogen and α2-antiplasmin with increased FVIII: C, von Willebrand factor, Fg degradation products and D dimers. Tissue plasminogen activator, plasminogen activator inhibitor and protein C varied. In L. achelous biological fluids, compounds with anticoagulant or procoagulant properties have been identified. In L. obliqua bristle extracts, mainly procoagulant activities have been identified. Subcutaneous injections of L. achelous crude extracts and a semipurified fraction reduce Fg, plasminogen and FXIII in rabbits. Intravenous injections of a very purified fraction of L. achelous in rabbits produce lysis of preformed thrombi, a decrease of Fg, plasminogen, α2-antiplasmin, FXIII and inhibition of postthrombolytic thrombus growth. Subcutaneous injections of L. obliqua bristle extracts prolong prothrombin time and activated partial thromboplastin time and reduce FXIII. Intravenous injections of crude bristle extract and a purified fraction of L. obliqua induce disseminated intravascular coagulation.
Identification of Anticoagulant Activities in Salivary Gland Extracts of Four Horsefly Species (Diptera, Tabanidae)
Authors: Kazimírová M. , Šulanová M. , Kozánek M. , Takáč P. , Labuda M. , Nuttall P.A.
Published: April 2003

Anticoagulant activities against the extrinsic and intrinsic coagulation pathways were identified in salivary gland extracts (SGE) prepared from four tabanids (Hybomitra muehlfeldi, Tabanus autumnalis, Haematopota pluvialis, Heptatoma pellucens). All extracts prolonged human plasma clotting time in a dose-dependent manner and inhibited thrombin activity in the chromogenic substrate assay. Horsefly SGE did not inhibit factor Xa. Partial purification of SGE proteins using reversed-phase high-performance liquid chromatography revealed species-specific differences in the elution profiles and range of fractions with anticoagulant activities.
Soluble Complement Receptor Type 1 Protects Rats from Lethal Shock Induced by Anti-Crry Antibody following Lipopolysaccharide Priming
Authors: Mizuno Masashi , Nishikawa Kazuhiro , Okada Noriko , Matsuo Seiichi , Okada Hidechika
Published: April 2003

Background: In rats primed with a trace amount of lipopolysaccharide (LPS), acute lethal shock is induced following the injection of monoclonal antibody against a membrane inhibitor of complement (anti-Crry). Administration of cobra venom factor to exhaust complement before the LPS priming can prevent the lethal reaction. Therefore, we evaluated whether soluble complement receptor type 1 (sCR1), which inhibits complement reaction, can interfere with lethal shock when administered after LPS priming or even after anti-Crry injection. Methods: sCR1 was administered intravenously before or after the administration of anti-Crry, and the effects on blood pressure and acute lethality were determined. Results: Administration of sCR1 could rescue rats from lethal shock even when it was administered after anti-Crry injection, which immediately causes a blood pressure decrease leading to lethal shock. Conclusion: sCR1 may be an effective treatment for acute shock involving complement activation.
Atopic Phenotype in Children Is Associated with Decreased Virus-Induced Interferon-α Release
Authors: Bufe Albrecht , Gehlhar Kirsten , Grage-Griebenow Evelin , Ernst Martin
Published: April 2003

Background: Interferon-α (IFN-α) production in humans is an early event in the nonspecific cellular response to viruses and mediates a wide range of antiviral and immunoregulatory activities. Little is known about the role of IFN-α in allergic disease. Methods: In the present study, we performed a retrospective comparative analysis of 88 children with and without an atopic phenotype for virus-induced IFN-α production in blood cultures. Results: We were able to demonstrate that patients with allergic asthma (aA) produced significantly lower amounts of virus-induced IFN-α than healthy children and patients with nonallergic asthma (naA). Furthermore, the number of eosinophils in atopic children as a marker for allergic inflammation correlated negatively with the IFN-α level in blood cultures. Additionally, we found differences between aA and naA patients with respect to the capacity to produce IFN-γ. Although atopy is thought to be associated with a Th2 cytokine response, in our study, IFN-γ release was not reduced in the allergic children. In contrast, patients with allergic rhinitis showed a significant increase in IFN-γ release compared to naA patients. Conclusions: In our study, an early atopic phenotype was related to a reduction in virus induced IFN-α release from blood cultures. Thus, after further prospective evaluation, the IFN-α level may serve as an additional in vitro marker for the definition of atopy in children.
Smouldering Mastocytosis: A Novel Subtype of Systemic Mastocytosis with Slow Progression
Authors: Valent Peter , Akin Cem , Sperr Wolfgang R. , Horny Hans-Peter , Metcalfe Dean D.
Published: April 2003

Systemic mastocytosis (SM) is a clonal disease that shows an either indolent or an aggressive clinical course. Utilizing established criteria, indolent SM can readily be discriminated from the rare aggressive subvariants of SM in most cases. In a small group of patients, however, clinical and laboratory parameters are indicative of slow progression without signs of aggressive disease or an associated hemopoietic neoplasm. These SM patients exhibit a high burden of mast cells, hypercellular marrow and organomegaly. Because of the ‘intermediate’ course and uncertain prognosis, these cases have been referred to as smouldering SM. In the present article, we discuss clinical and laboratory findings in smouldering SM and review the current literature. In addition, the pathophysiology of this novel subtype of SM is discussed.
Perinatal Maternal Food Restriction Induces Alterations in Hypothalamo-Pituitary-Adrenal Axis Activity and in Plasma Corticosterone-Binding Globulin Capacity of Weaning Rat Pups
Authors: Léonhardt Marion , Lesage Jean , Dufourny Laurence , Dickès-Coopman Anne , Montel Valérie , Dupouy Jean-Paul
Published: April 2003

We investigated the effects of perinatal maternal malnutrition on the hypothalamo-pituitary-adrenal (HPA) axis activity in both basal and stressful conditions in newborn rats at weaning. Mothers from the control group were fed ad libitum. Mothers exposed to food restriction received 50% (FR50) of the daily intake of pregnant dams during the last week of gestation (Pre group), lactation (Post group) or both periods (PP group) in order to compare the long-term effects of gestational and/or lactational restriction. FR50 reduced the body growth of pups from the Post and PP groups as soon as day 11 until day 21 after birth. At weaning, pups of the Post and PP groups showed reduced adrenal, thymus and liver weights. Although the plasma adrenocorticotropic hormone (ACTH) level was reduced in pups, FR50 affected neither corticotropin-releasing hormone expression and peptide synthesis in the hypothalamus nor proopiomelanocortin expression in the adenohypophysis. Basal circulating levels of corticosterone were not markedly affected by FR50, but free corticosterone concentration was increased in the PP group. Plasma corticosterone-binding globulin (CBG) was decreased in newborns from both the Post and PP groups. Mineralocorticoid receptor gene expression was significantly increased in both CA1 and CA3 hippocampal areas in the PP group. Glucocorticoid receptor gene expression was increased in CA1, CA2 and dentate gyrus hippocampal areas in the Pre group, as well as in CA1, CA3 and DG areas in the Post group. The ether inhalation-induced plasma ACTH increase was weaker in pups from the Post and PP groups. Similarly, the ether inhalation-induced plasma corticosterone increase returned to basal levels in the Post group, or to weaker values than baseline in the PP group 90 min after this stressful procedure. The present work suggests that maternal food restriction during the perinatal period (gestation and lactation) or during lactation only reduces the postnatal somatic growth of pups and disturbs the activity of the HPA axis at weaning under both resting and stress conditions. A reduction in the plasma CBG-binding capacity, associated with a probable increase in hippocampal corticosteroid receptors, could reinforce glucocorticoid-mediated negative feedback and shorten stress-induced activation of the HPA axis in pups at weaning.
Hypophyseal-Portal Somatostatin (SRIH) and Jugular Venous Growth Hormone Secretion in the Conscious Unrestrained Ewe
Authors: Veldhuis Johannes D. , Fletcher Terry P. , Gatford Kathryn L. , Egan Adrian R. , Clarke Iain J.
Published: April 2003

Somatostatin (SRIH) release into hypophyseal portal blood varies reciprocally with growth hormone (GH) pulse generation in the male rat. However, few studies have directly evaluated this relationship in the female of any species. To address this issue, we carried out intensive (5 min) and extended (240 min) simultaneous monitoring of hypophyseal portal SRIH and internal jugular GH secretion in 7 unanesthetized ewes. Bihormonal synchrony was assessed by three statistically independent but complementary analyses: (i) cross-approximate entropy (X-ApEn) analysis to appraise the conditional regularity of SRIH/GH release patterns; (ii) cross-correlation analysis of paired sample SRIH and GH release rates, and (iii) probability analysis of random versus nonrandom SRIH and GH discrete pulse concordance. From a one-variable perspective, ApEn analysis documented consistently more irregular patterns of SRIH than GH release (94 ± 4.3 and 72 ± 8.1%, respectively, of the mean irregularity of 1,000 individual random-shuffled cognate series, p = 0.034). From a two-variable perspective, X-ApEn analysis revealed a nearly mean random relationship between SRIH and GH release patterns (group mean ± SEM, 94 ± 4.5% of the mean asynchrony of 1,000 randomly shuffled SRIH/GH pairs). Cross-correlation analysis disclosed highly variable linkages between SRIH and GH secretion; viz, negative cross-correlations in 5 sheep, positive relationships in 4, and both positive and negative SRIH/GH associations in 2 animals, wherein changes in SRIH secretion either preceded or followed those of GH. Peak detection by model-free cluster analysis quantified a total of 28 SRIH and 31 GH release episodes. Corresponding interpulse intervals (min) were comparable (37 ± 4 (SRIH) and 43 ± 12 (GH)), but the mean fractional (%) amplitude of SRIH peaks was 3.5-fold lower (60 ± 10%) than that for GH (225 ± 50%) (p = 0.024). Pulse-concordance probability testing showed that discrete peaks of SRIH and GH secretion coincided only 33% of the time, although this value exceeded chance expectation (p < 10–4). In summary, the present analysis applies intensive (5 min) and extended (240 min) simultaneous sampling of hypophyseal-portal and jugular venous blood to quantitate the degree of coordinate SRIH and GH secretion in the unanesthetized ovariectomized ewe. Thereby, we unmask highly irregular SRIH release patterns, and nearly random SRIH and GH associations. We conclude that, to the extent that in vivo sampling reflects physiological SRIH/somatotrope activity, the female sheep maintains complex time-varying interactions between SRIH and GH release.
Direct Visualization of Internalization and Intracellular Trafficking of Dopamine-Releasing Protein-36aa
Authors: Smith Sean , Ramirez Victor D.
Published: April 2003

Dopamine-releasing protein (DARP) is a potent regulator of dopamine (DA) release known to be involved in the development of rat catecholaminergic systems. In the present study, we examined the internalization and transport of DARP-36aa, a synthetic peptide derived from the N-terminal sequence of DARP, in rat C6 glioma cells. A colloidal gold DARP-36aa conjugate (DARP-36aa: AU) and biotinylated DARP-36aa were employed to visualize internalization and intracellular transport of DARP-36aa. Electron microscopy demonstrated that DARP-36aa: AU was rapidly incorporated into C6 glioma cells. Internalization via clathrin-coated pits and vesicles was clearly observed followed by transport and sorting of DARP-36aa: AU into multivesicular bodies, tubulo-vesicular endosomes, and lysosomes. Internalization of DARP-36aa: AU was also examined in primary mesencephalic cell cultures where a similar pattern of internalization and transport via clathrin-coated pits and vesicles was observed. Fluorescence microscopy using a biotinylated DARP-36aa/avidin-rhodamine conjugate revealed that DARP-36aa is diffusely distributed on the plasmalemma prior to internalization at 4°C. Following a 30-min incubation at 37°C DARP-36aa was concentrated in the cytosol, particularly in areas surrounding cellular projections and the perikaryon. Immunocytochemical studies employing biotinylated DARP-36aa and an anti-clathrin heavy chain antibody demonstrated that DARP-36aa and clathrin colocalize during DARP-36aa internalization. We also observed a marked increase in tyrosine phosphorylation of a 45-kD protein in response to DARP-33a stimulation in C6 glioma cells. Genistein, a specific tyrosine kinase inhibitor, significantly inhibited DARP-36aa: AU internalization and transport in C6 glioma cells. These findings suggest that tyrosine kinase activity may result in DARP-36aa receptor-mediated endocytosis.
Circadian Rhythms of Dopamine and Dihydroxyphenyl Acetic Acid in the Mouse Striatum: Effects of Pinealectomy and of Melatonin Treatment
Authors: Khaldy Hoda , León Josefa , Escames Germaine , Bikjdaouene Leila , García Joaquín José , Acuña-Castroviejo Darío
Published: April 2003

The existence of dopamine (DA)-melatonin (aMT) relationships is well documented in several brain areas of the mammalian central nervous system such as the retina and hypothalamus or the nigrostriatal system. For instance, aMT tempers 1 methyl-4-phenyl-1,2,3,6-tetrahydropyridine-induced nigrostriatal damage in C57BL/6 mice. In this mouse strain however, rhythmic production of aMT and its possible interaction with striatal DA is still unclear. In the present work we investigated circadian variations in pineal production of aMT and striatal DA levels in C57BL/6 mice. Effects of pinealectomy and aMT administration were also assessed. Intact, pinealectomized and pinealectomized + aMT-treated mice and their respective control groups were sacrificed at six different times during the 24-hour period. In control animals, aMT displayed a circadian rhythm with a narrow peak at midnight. The peak of aMT coincided with the nadir of the DA rhythm present in the striatum. Shortly after the decrease of DA levels, an increase in 3,4-dihydroxyphenylacetic acid (DOPAC), the main DA metabolite, was observed. The rhythmic changes of DA and DOPAC levels in the striatum were blunted by pinealectomy, whereas administration of aMT (0.1–10 mg/kg) during 6 days to pinealectomized mice restored the rhythms in a dose-dependent manner. Striatal levels of 3-methoxytyramine and homovanillic acid did not change during the 24-hour cycle. The serotonergic system, assessed by the determination of 5-hydroxytryptamine and 5-hydroxyindole-3-acetic acid concentration in striatum, did not show significant time-dependent changes in control animals and was not affected by pinealectomy or aMT treatment. These data substantiate the existence of a link between pineal function, melatonin secretion and DA circadian rhythm in the mouse striatum.
Phase I Study of Vinorelbine and Carboplatin Combination in Patients with Taxane and Anthracycline Pretreated Advanced Breast Cancer
Authors: Kosmas C. , Agelaki S. , Giannakakis T. , Mavroudis D. , Kouroussis Ch. , Kalbakis K. , Papadouris S. , Souglakos J. , Malamos N. , Georgoulias V.
Published: April 2003

Aim: To define the maximum tolerated dose (MTD) and the dose-limiting toxicities (DLTs) of the carboplatin-vinorelbine combination in pretreated patients with advanced breast cancer. Patients and Methods: Patients with histologically confirmed metastatic breast cancer relapsing or progressing after prior taxane and anthracycline containing chemotherapy were enrolled. Cohorts of 3–6 patients were treated at successive dose levels (DLs) with escalated doses of carboplatin [range, area under the curve (AUC) 4–6] on day 1 and vinorelbine (range, 20–35 mg/m2) on days 1 + 8 recycled every 28 days. Results: Twenty-seven patients with a median age of 58 years and performance status (WHO) of 0–2 were treated at 6 DLs. All patients were assessable for toxicity and 20 for response. DLT was reached at carboplatin 6 AUC and vinorelbine 35 mg/m2, and therefore, this was considered as the MTD. Prophylactic G-CSF administration could not allow further dose escalation. The recommended dose for further phase II testing was defined at carboplatin 6 AUC on day 1 and vinorelbine 30 mg/m2 on days 1 and 8. Among 98 administered treatment cycles 41 (42%) and 7 (7%) were complicated with grades 3 and 4 neutropenia and thrombocytopenia, respectively. Nonhematologic toxicities included grade 2 peripheral neuropathy in 3 cycles and grades 2 and 3 fatigue in 32 (32%). Conclusion: The present study determined the feasibility of the combination of carboplatin at AUC 6 (day 1) and vinorelbine at 30 mg/m2 (days 1 and 8 ) without G-CSF support in patients with taxane and anthracycline pretreated advanced breast cancer. Phase II studies at these doses should follow in order to determine the activity of the regimen.
Phase I Study of Gemcitabine and Liposomal Doxorubicin in Relapsed Ovarian Cancer
Authors: D’Agostino Giuseppe , Ferrandina Gabriella , Garganese Giorgia , Salerno M. Giovanna , Lorusso Domenica , Farnetano M. Giuseppina , Mancuso Salvatore , Scambia Giovanni
Published: April 2003

Twenty-three patients were enrolled in a phase I study conducted to determine the maximum tolerated doses (MTD) of combined liposomal doxorubicin (CAE) and gemcitabine (GEM) in relapsed ovarian cancer patients. A total of 82 courses are evaluable, with a median number of three cycles administered per patient (range 2–8). GEM was administered on days 1 and 8 by 30-min intravenous infusion immediately after CAE given by 60-min intravenous infusion on day 1; cycles were repeated every 21 days. The starting doses were CAE 20 mg/m2 and GEM 600 mg/m2. Following dose levels were 20/800; 20/1,000; 30/800; 30/1,000; 35/800, and 35/1,000 for CAE and GEM, respectively. The MTD was reached at dose level 5, with febrile neutropenia and thrombocytopenia as dose-limiting toxicities. After the MTD, granulocyte-colony stimulating factor was administered in 15% of cycles. Non-hematological toxicity was mild and manageable. All patients are so far evaluable for response. Among them, 5 partial responses (21.7%; 95% confidence interval, CI: 4.9–38.5), 5 disease stabilizations (21.7%, 95% CI: 4.9–38.5) and 13 progressions (56.6%, 95% CI: 36.4–76.8) have been registered. These results warrant further research in a phase II study.
ARHI/NOEY2 Inactivation May Be Important in Breast Tumor Pathogenesis
Authors: Hisatomi Hisashi , Nagao Kumi , Wakita Kazuyuki , Kohno Norio
Published: April 2003

Allelic loss frequently occurs on the short arm of chromosome 1 in human breast carcinoma, suggesting that the ARHI/NOEY2 gene, an imprinted putative tumor suppressor gene, is involved in the pathogenesis of the tumor entity. To clarify the clinical importance of ARHI/NOEY2 mRNA in breast cancer, we studied whether ARHI/NOEY2 inactivation might contribute to tumors arising in the breast. An ARHI/NOEY2 message was detected by real-time PCR analysis in all noncancerous breast tissues, but was not detected in 2 of 26 breast cancer tissue samples. In 10 of 26 breast cancer tissue samples ARHI/NOEY2 mRNA was substantially reduced. ARHI/NOEY2 expression was lost or markedly reduced in 12 of 26 (46.15%) breast cancer tissue samples. In summary, we conclude that decreased ARHI/NOEY2 mRNA expression may play an important role in the pathogenesis of breast cancer.
Comparisons of Risk Factors and Visual Field Changes between Juvenile-Onset and Late-Onset Primary Open-Angle Glaucoma
Authors: Ko Yu-Chieh , Liu Catherine Jui-Ling , Chou Joe Ching-Kuang , Chen Mei-Ru , Hsu Wen-Ming , Liu Jorn-Hon
Published: April 2003

Purpose: To identify the differences in risk factors and visual field (VF) changes between juvenile primary open-angle glaucoma (JOAG) and late-onset chronic open-angle glaucoma (COAG). Methods: The demographic and presenting clinical data of 27 JOAG and 30 COAG patients were retrospectively reviewed. Comparisons between the two groups were performed using Mann-Whitney U test, Wilcoxon signed-rank test, and Fisher’s exact test. Results: A family history of glaucoma (37%) and a history of steroid usage (14.8%) were identified in JOAG patients only. The JOAG patients had a longer axial length (p < 0.001) and more often a myopic refractive state (p < 0.001) than the COAG patients. Patients with COAG showed a deeper (p = 0.016) and a more extensive (p = 0.008) defect in the superior than in the inferior hemifield, as well as a deeper (p = 0.016) and a more extensive (p = 0.001) defect in the superior than in the inferior arcuate area, while JOAG patients showed symmetric VF defects between the superior and inferior hemifields. Purely diffuse VF depression is more common in JOAG than in COAG patients (p = 0.03). Conclusions: JOAG patients demonstrated more axial myopic changes than patients with COAG as well as a pattern of superior-inferior symmetric VF defects. Axial myopia may play a critical role in the pathogenesis of JOAG.
Optic Neuropathy from Folic Acid Deficiency without Alcohol Abuse
Authors: Hsu Cynthia T. , Miller Neil R. , Wray Misty L.
Published: April 2003

A 47-year-old woman with a 2-month history of bilateral progressive visual loss was found to have a bilateral retrobulbar optic neuropathy. Her serum vitamin B12 concentration and hemoglobin level were normal, but her serum folic acid concentration was decreased. The patient had a minimal alcohol intake and moderate tobacco use that had been unchanged for over 20 years; however, she had markedly altered her diet 4 years earlier in the setting of clinical depression. After treatment with oral folic acid and diet modification without change in her tobacco or alcohol use, the patient’s visual function returned to normal. This case supports the role of folic acid deficiency as an important cause of some cases of nutritional optic neuropathy.
Typical Ultrasound Biomicroscopic Findings Seen in Ocular Hypotony
Authors: Roters Sigrid , Engels Bert F. , Szurman Peter , Krieglstein Günter K.
Published: April 2003

Detailed imaging of the ciliary body region by means of ultrasound biomicroscopy can provide information that is crucial for further treatment whereas clinical evaluation and 10-MHz standard B scan are less helpful. Background: To evaluate the suitability of ultrasound biomicroscopy (UBM) for elucidating the causes of ocular hypotony. Patients and Methods: A retrospective chart review was performed to determine the usefulness of UBM in the evaluation of ocular hypotony. UBM, B scan, and a detailed clinical examination had been performed. Sixty patient charts with prolonged ocular hypotony of different causes were reviewed from January 1994 to December 1998. The information obtained by the UBM was classified into three groups: ‘diagnostic’ (group 1), ‘helpful’ (group 2), ‘not helpful’ (group 3). Results: Of the 60 eyes, 45 UBM examinations (75%) were in group 1, 9 eyes (15%) were assigned to group 2 and 6 eyes (10%) were in group 3. The duration of hypotony had no influence. Conclusion: In cases of ocular hypotony, the cause was related to ciliary body pathologies in 80%. Only UBM could distinguish tractional from dehiscence ciliary body detachment which required a different management approach. UBM with its high resolution imaging of the anterior segment – including the ciliary body – is highly suitable for diagnostic clarification.
Re-Worsening Factor after Successful Vitrectomy for Diabetic Retinopathy: Optic Disc Fibrovascular Proliferation and Macular Disease
Authors: Sakamoto Taiji , Fujisawa Kimihiko , Kinukawa Naoko , Ishibashi Tatsuro , Inomata Hajime
Published: April 2003

Purpose: To investigate the factors that influence the visual-changing pattern in proliferative diabetic retinopathy even after successful vitrectomy. Methods: One hundred and forty-seven consecutive eyes were retrospectively reviewed for 6–48 (average 20) months, and were divided into the following 4 groups based on their changing pattern of vision: group A, the visual acuity improved postoperatively and maintained the maximal corrected vision throughout the observation period (n = 49); group B, the visual acuity improved postoperatively but deteriorated thereafter (n = 68); group C, the visual acuity remained the same as before operation (n = 17), and group D, the visual acuity deteriorated immediately after vitrectomy (n = 13). Various issues including systemic conditions, blood tests, preoperative ocular findings, the operative procedures and postoperative complications were reviewed based on the patient records. These issues were analyzed by Spearman’s rank correlation, χ2 test, an analysis of variance (ANOVA) and the Kruskal-Wallis test. Finally, the discriminate factors between groups A and B were examined by a stepwise logistic regression analysis. Results: The following tendencies were observed in all 4 groups: younger patients tended to show a better visual-changing pattern (p = 0.02); patients with younger age at diabetes onset had a better visual-changing pattern after vitrectomy (p = 0.001), and a lower hemoglobin (Hb) A1c level is associated with a better visual changing pattern (p = 0.017). Preoperative rubeosis and macular detachment were frequently found in groups C and D, as well as postoperative rubeosis, vitreous bleeding and retinal detachment. Finally, a stepwise logistic regression analysis showed both fibrovascular proliferation (p = 0.016) from the optic disc and postoperative macular disease (p = 0.0009) to be significant factors for differentiating group A from group B. Conclusions: In addition to the factors which have already been indicated to affect the visual outcome of a vitrectomy, preoperative findings such as optic disc fibrovascular proliferation and postoperative macular disease were found to affect the visual-changing pattern after a successful vitrectomy. The optimal timing of surgery is very important not only in order to obtain good visual acuity but also to maintain good vision even after a successful vitrectomy.