bluuebunny/crossref_metadata_2025 huggingface dataset:
Proteases from Vipera lebetina Venom Affecting Coagulation and Fibrinolysis
Authors: Siigur J. , Aaspõllu A. , Tõnismägi K. , Trummal K. , Samel M. , Vija H. , Subbi J. , Siigur E.
Published: April 2003

Our studies of the venom from the Levantine viper Vipera lebetina have demonstrated the existence of both coagulants and anticoagulants in the same venom. We showed that V. lebetina venom contains: (1) proteases that degrade fibrinogen, but not fibrin; (2) fibrinolytic enzyme (lebetase); (3) factor X activator (VLFXA); (4) factor V activator (VLFVA). Fibrinolytic enzyme and VLFXA are metalloproteases; the other studied enzymes are serine proteases. α-Fibrinogenase has no homolog among known serine proteases. β-Fibrinogenase is a typical thermostable arginine esterase that hydrolyzes esters and amides of arginine and attacks the β-chain of fibrinogen. Lebetase is a direct-acting fibrinolytic zinc metalloendopeptidase related in amino acid sequence to reprolysins. We used the matrix-assisted laser desorption/ionization time-of-flight mass spectrometry technique for the recovery and identification of peptides released by protease hydrolysis and for the detection of human factor X cleavage products after VLFXA hydrolysis. VLFXA cleaves the Arg52-Ile53 bond in the heavy chain of human factor X and the Arg226-Val227 bond in human factor IX precursor; VLFVA cleaves Arg1545-Ser1546 in factor V.
Lebecetin, a C-Lectin Protein from the Venom of Macrovipera lebetina That Inhibits Platelet Aggregation and Adhesion of Cancerous Cells
Authors: Sarray Sameh , Srairi Najet , Luis Jose , Marvaldi Jacques , El Ayeb Mohamed , Marrakchi Naziha
Published: April 2003

A novel C-lectin protein, lebecetin, was purified and characterized from the venom of Macrovipera lebetina. It is a disulfide-linked heterodimer of 15 and 16 kD. The subunits are homologous to each other and to the other snake venom proteins of the C-type (Ca2+-dependent) lectin superfamily. Lebecetin shows a potent inhibitory effect on whole blood and washed platelets induced by different agonists. It inhibits the agglutination of human fixed platelets in the presence of ristocetin. Lebecetin also interferes with the adhesion of IGR39 melanoma and HT29D4 adenocarcinoma cells. These two lines adhere to lebecetin used as matrix. Lebecetin is also able to strongly reduce IGR39 and HT29D4 cell adhesion to fibrinogen and laminin, but not to fibronectin and collagen types I and IV, respectively. Adhesion properties of lebecetin may thus involve integrin receptors.
Diagnostic Uses of Snake Venom
Authors: Marsh Neville A.
Published: April 2003

Snake venom toxins are invaluable for the assay of coagulation factors and for the study of haemostasis generally. Thrombin-like enzymes (SVTLE) are used for fibrinogen and fibrinogen breakdown product assays as well as detecting dysfibrinogenaemias. Since SVTLE are not inhibited by heparin, they can be used for assaying antithrombin III in samples containing heparin. Snake venom prothrombin activators are utilised in prothrombin assays, whilst Russell’s viper venom (RVV) can be used to assay clotting factors V, VII, X and lupus anticoagulants (LA). Activators from the taipan, Australian brown snake and saw-scaled viper have also been used to assay LA. Protein C (PC) and activated PC (APC) resistance can be measured by means of RVV, ProtacTM (from Southern copperhead snake venom) and STA-Staclot (from Crotalus viridis helleri) whilst von Willebrand factor can be studied with BotrocetinTM(Bothrops jararaca). Finally, snake venom C-type lectins and metalloproteinase disintegrins are being used to study platelet glycoprotein receptors and show great potential for use in the routine coagulation laboratory.
Snake Venom Activators of Factor X: An Overview
Authors: Tans Guido , Rosing Jan
Published: April 2003

Activators of blood coagulation factor X have been described in the venom of many snake species belonging to the genus Viperidae and Crotalidae as well as from a few Elapid species. Based on the structural and functional properties of purified activating principles, factor X activators are either metalloproteases or serine proteases. The best known activator is RVV-X from Russell’s viper (Daboia russelli), a metalloprotease consisting of a heavy chain containing the catalytic domain and two light chains which share homology with C-type lectins and which are thought to exert a regulatory function in the Ca2+-dependent activation of factor X. This activator is also one of the best examples of the use of exogenous activators in coagulation research and in addition it is used in many diagnostic research kits. In this paper, an overview is given of the structural and functional properties of snake venom factor X activators thus far described in the literature.
Effects of Lopap on Human Endothelial Cells and Platelets
Authors: Chudzinski-Tavassi A.M. , Schattner M. , Fritzen M. , Pozner R.G. , Reis C.V. , Lourenço D. , Lazzari M.A.
Published: April 2003

Severe consumption coagulopathy has been detected in rats after Lopap (a prothrombin activator from Lonomia obliqua caterpillar bristles) infusion and in humans after accidental contact with L. obliqua bristles. However, platelet count and antithrombin (AT) levels were only modestly affected, suggesting that a different form of blood coagulation activation may be involved in this hemorrhagic syndrome. Here we describe that Lopap had no effect on aggregation of washed human platelets induced by several agonists, suggesting that it might not impair platelet function in vivo. AT was able to inhibit the amidolytic activity of thrombin generated by incubation of Lopap with prothrombin in a purified system, which may be different from that generated by the prothrombinase complex in vivo. The surface expression of both ICAM-1 and E-selectin but not of VCAM-1 was upregulated by Lopap in cultured HUVEC, suggesting that it may behave differently from other mediators, such as thrombin and tumor necrosis factor-α.
A Cross-Sectional Study of Dehydroepiandrosterone Sulfate in Prepubertal Children with Myelomeningocele
Authors: Trollmann Regina , Langhans Beate , Strehl Elisabeth , Wenzel Dieter , Dörr Helmuth G.
Published: April 2003

Objective: To assess biochemical characteristics of adrenarche in patients with myelomeningocele (MMC), we examined serum levels of dehydroepiandrosterone sulfate (DHEAS) in prepubertal MMC patients. Patients and Methods: The study included a total of 54 prepubertal patients with MMC and shunted hydrocephalus: 13 patients (2 m, 11 f; aged 4.6–10.1 years, mean 8.1 ± 0.4) with isolated pubarche (Tanner stage PH 2–4, B1 or testes volume ≤3 ml) and 41 prepubertal MMC patients without pubarche (17 m, 24 f; aged 2.0–11.9 years; mean 6.8 ± 2.5). DHEAS levels were measured directly by chemiluminescence immunoassay (Nichols, USA). Auxological data (supine length, body mass index (BMI), arm span) and bone age (BA) were recorded. Results(mean ± SD): Basal DHEAS levels correlated with chronological age (CA) (r = 0.32, p < 0.05), BA (r = 0.65, p < 0.01; n = 23), BMI (r = 0.54, p < 0.01) and pubic hair stage (PH1 vs. PH2–4, r = 0.49, p < 0.01). 10/11 patients aged 2–4 years had DHEAS levels in the normal range, whereas 18/40 (45.0%) of the 5- to 9-year-old patients showed elevated levels (>+2 SDS). Ten patients with isolated pubarche (10/13; 2 m, 8 f; CA 8.3 ± 1.5 years) and 9 patients without pubarche (9/41; 6 m, 3 f; CA 6.9 ± 2.1 years) had elevated DHEAS levels (+6.34 and +4.05 SDS, respectively). The values correlated with BA/CA ratio (p < 0.05, n = 23). There was a trend to higher BMI SDS levels in patients with elevated DHEAS levels. Conclusion: Our data show an early and increased activation of adrenal androgen secretion in MMC patients.
Psychological Evaluation of Young Women after Medical Treatment for Central Precocious Puberty
Authors: Baumann D.A. , Landolt M.A. , Wetterwald R. , Dubuis J.M. , Sizonenko P.C. , Werder E.A.
Published: April 2003

Objective: This study aimed at the evaluation of the subjective experience and long-term behavioral and psychological effects of precocious puberty (PP). Methods: 19 female patients who had been treated with GnRH agonists participated in a semistructured interview and completed two standardized checklists. Their parents completed the Child Behavior Checklist (CBCL). Results: The CBCL yielded significantly elevated Internalizing and Total Behavior Problem scores. An elevated risk was found for patients with short adult stature and a relatively late onset of PP. The latter tended to neuroticism, to accentuation of their physical appearance, and felt significantly more insecure than age-related non-PP girls. Conclusion: Our findings suggest that PP can lead to specific behavioral problems, and that patients with a risk factor may need psychosocial support.
Early Fractures and Occult Hyperthyroidism: McCune-Albright Syndrome?
Authors: Di Candia Stefania , Weber Giovanna , Mora Stefano , de Pellegrin Maurizio , Chiumello Giuseppe
Published: April 2003

McCune-Albright syndrome (MAS) is a sporadic disease characterized by fibrous bone dysplasia, café-au-lait spots and hyperfunctional endocrinopathies. We report a 2.5-year-old child with MAS with an early and nonclassic onset. He was admitted to our attention for frequent fractures without clinical signs of endocrinopathies, found to have asymptomatic, nonautoimmune hyperthyroidism. The diagnosis of MAS was based on RX and MR imaging associated with hyperthyroidism. It is not clear if there was a correlation between the severity of bone disease and the presence of thyroid disorder. At the moment no standard treatment exists for bone fibrous dysplasia and hyperthyroidism in children before the age of 6 years.
Measurements of Urinary Leptin and Capillary Leptin: Alternative Tools for the Assessment of the Leptin Status?
Authors: Dimitriou Triantafillia , Huesemann Sabine , Neubert Annette , Remer Thomas
Published: April 2003

Objectives: The aim of this study was to check whether leptin is reliably measurable in urine samples of children, adolescents, and adults and to examine whether capillary leptin measurements can be utilized as an alternative tool to assess the leptin status. Methods: Two studies were performed. In both studies, leptin was quantified by an ultrasensitive and highly specific enzyme immunoassay (ELISA; R & D Systems). Anthropometric measures were taken from all study subjects, and body fat was calculated using skinfold thickness measurements. In study 1, leptin was analyzed in 24-hour urine samples of 155 healthy children and adolescents and 26 healthy adults after a methodological modification of the assay necessary for urine analysis. In study 2, venous and capillary blood samples were collected in 26 healthy adults within 10 min on the same day. Results: After adapting the assay system to urine matrix, the detection range was 20–160 pg/ml. Only in 2 of 181 urine samples reproducibly measurable urinary leptin concentrations in the lowest detection range were found. In study 2, a close correlation was found between log capillary and log venous leptin concentrations (r = 0.98, p < 0.001) and between log capillary as well as log venous leptin levels and percent body fat (r = 0.86, p < 0.001). Conclusions: Our results based on one of the most specific and sensitive ELISAs currently available show that leptin is generally undetectable in the urine from healthy children, adolescents and adults. Thus, urinary leptin excretion cannot be used as a noninvasive marker of the leptin status. Our findings in healthy adults show that the merely moderately invasive determination of capillary leptin allows a reliable assessment of the individual leptin status and may be used instead of venous leptin as a biochemical indicator of body fatness.
Alteration of Endothelin-1 Concentration in STZ-Induced Diabetic Rat Nephropathy
Authors: Itoh Yasutoshi , Nakai Akira , Kakizawa Hiroaki , Makino Masaki , Fujiwara Kentaro , Kobayashi Takako , Kato Taiya , Nagata Mutsuko , Oda Naohisa , Katsumata Hideki , Nagasaka Akio , Itoh Mitsuyasu
Published: April 2003

Background: Recently, an endothelin (ET-1) with a potent vasoconstrictive activity and stimulative activity of vascular muscular cell growth was discovered and blood ET-1 levels were higher in diabetic patients than in healthy subjects, suggesting that high ET-1 levels assist development and progression of diabetic microangiography. Methods: We examined renal function, and serum and tissue ET-1 levels in streptozotocin (STZ)-induced diabetic rats treated with a prostaglandin (PG) I2 derivative to investigate the effect of PGI2 in diabetic vascular disturbance. Results: Renal weight, urinary albumin, urinary N-acetyl-β,D-glucosaminidase (NAG) and serum ET-1 levels increased in STZ-induced diabetic rats, and a tendency to increase in renal tissue ET-1 levels was observed. Furthermore, electron-microscopic findings in the kidneys showed mesangial cell proliferation and mesangial matrix expansion which might be caused by diabetic nephropathy. The PGI2 derivative reduced urinary albumin and NAG levels in STZ-induced rats. It was considered, therefore, that the PGI2 derivative is effective in diabetic nephropathy. As the PGI2 derivative also reduced renal tissue ET-1 levels, improvement of diabetic nephropathy partially was considered to result from the reduction of renal tissue ET-1 levels. Conclusion: In STZ-induced rats, increased serum ET-1 levels and a tendency to increase in renal tissue ET-1 levels were associated with increases in urinary albumin and NAG levels, and these levels were decreased by a PGI2 derivative. These findings suggested that increased ET-1 concentrations assist development and progression of diabetic nephropathy, especially diabetic microangiopathy, and the PGI2 derivative may be effective for inhibition of diabetic microangiopathy mediated by reduction of ET-1 concentrations.
Growth Hormone Therapy in the Glucocorticosteroid-Dependent Child: Metabolic and Linear Growth Effects
Authors: Mauras Nelly
Published: April 2003

Pharmacological doses of glucocorticosteroids given chronically are associated with a variety of negative side effects which impact the prolonged use of these potent anti-inflammatory agents. They have catabolic effects on protein, resulting in poor tissue healing, an increased incidence of infections and accelerated bone loss. Insulin resistance to both hepatic and peripheral tissues is a common consequence of chronic steroid use, leading at times to impaired carbohydrate metabolism. Steroids affect both the release and the effects of growth hormone (GH) at the target sites, hence becoming functional GH antagonists. When administered to growing children the side effects of glucocorticosteroid treatment are further compounded by a potent and significant suppression of linear growth. Ample experimental and clinical data support a role for GH therapy in counteracting some of the effects of glucocorticosteroids. Using isotope dilution methods we have previously shown that both GH and insulin-like growth factor (IGF)-I can decrease the protein wasting effects of prednisone administration in man. IGF-I has also been shown to enhance type I collagen formation in hydrocortisone-treated human osteoblasts. GH (through IGF-I) significantly enhances linear growth; thus, in states of ‘functional’ GH deficiency, such as that observed in chronic steroid use, GH may also have a potentially beneficial effect. Studies in children on chronic prednisone doses with cystic fibrosis, chronic renal failure or juvenile rheumatoid arthritis have all shown beneficial effects on linear growth after prolonged GH therapy. Data from a recent study of ours using GH in children with steroid-dependent inflammatory bowel disease showed that GH treatment was associated with increased lean body mass, decreased adiposity and increased linear growth. Marked increases in IGF-I concentrations and in kinetic measures of bone calcium accretion (using calcium tracers) were also observed, without any deterioration of disease activity scores or carbohydrate tolerance. In conclusion, GH therapy may play a role in the treatment of children on chronic steroids both as a growth promoting agent and as an anabolic agent on whole body protein and bone. Longer term studies will be needed to better define the safety and efficacy of this approach.
From Fixed to Individual Dosing in Growth Hormone-Deficient Adults
Authors: Vance Mary Lee
Published: April 2003

Initial research studies of growth hormone (GH) replacement in adults employed what is now considered to be high doses of GH, from 12 to 25 µg/kg per day. While the beneficial effects of GH were demonstrated, there was a considerable incidence of side effects, most commonly edema and arthralgia. These side effects abated with a dose reduction. More recently, the Growth Hormone Research Society recommended initiating GH replacement at a dose of 0.3 mg/day irrespective of total or ideal body weight. The United States Food and Drug Administration approved pharmaceutical company package inserts that recommended beginning with a dose of 6 µg/kg per day with a maximum dose of 25 µg/kg per day and limiting the maximal dose to 12.5 µg/kg per day in adults over the age of 35 years. These should be considered as guidelines.
Testosterone Deficiency and Replacement
Authors: Howell Simon , Shalet Stephen
Published: April 2003

In the human male, testosterone is the major circulating androgen. More than 95% of circulating testosterone is secreted by the testis with a production rate of 6–7 mg/day. The clinical effects of androgens are numerous, and testosterone deficiency is associated with a number of clinical abnormalities. Overt hypogonadism results in reductions in bone mineral density, alterations in body composition and effects on mood, aggressive behaviour, cognitive function, sexual function and several factors important for cardiovascular risk. Androgen replacement in this context is clearly beneficial, and numerous studies have demonstrated improvements in bone and muscle mass, reductions in body fat, and positive effects on quality of life following treatment. The benefits of therapy in men with milder degrees of hypogonadism, and elderly men with ‘physiological’ testosterone deficiency, are less clear-cut, and the appropriate biochemical cut-off below which replacement should be offered has not been clearly defined. Several options are available for androgen replacement in adult men. Oral testosterone, intramuscular injections, subcutaneous implants and transdermal therapy have all been used. Each mode of delivery has advantages and drawbacks and the choice between them will often depend on patient reference. Recent advances include the development of longer-acting intramuscular preparations, which offer more stable androgen levels with fairly infrequent injections, and testosterone gel which appears to provide transdermal replacement without a high incidence of skin reactions. This article will examine the evidence concerning the impact of male hypogonadism and the response to androgen therapy. The question of who to treat will be addressed with particular reference to mild hypogonadism and hypogonadism in the elderly. Finally, an overview of the different modes of replacement therapy will be presented.
Pathophysiology of Chronic Urticaria
Authors: Greaves Malcolm W.
Published: April 2003

Chronic urticaria includes several different subsets with distinct pathophysiologies, and with important implications for investigation and treatment. Chronic ‘idiopathic’ urticaria represents a special challenge, which, until recently, was not taken up by dermatological or immunological investigators. However, it has now emerged that at least 30% of patients possess histamine-releasing autoantibodies against FcΕR1, or less commonly IgE itself. These autoantibodies are causative. Recent work implicates complement activation in most cases. Functional (i.e. histamine releasing) autoantibodies are specific to chronic urticaria. However, immunoreactive (non-histamine-releasing) anti-FcΕR1 autoantibodies can be found in sera of patients with physical urticarias and with autoimmune connective tissue and bullous diseases. The reason for the occurrence of this disease type in some individuals but not others is unclear. One possibility is the development, in genetically predisposed persons, of autoantibodies by molecular mimicry – perhaps against lipopolysaccharide of Helicobacter pylori, an organism frequently infecting the upper gastrointestinal tract of chronic urticaria patients.
Kit and c-kit Mutations in Mastocytosis: A Short Overview with Special Reference to Novel Molecular and Diagnostic Concepts
Authors: Féger Frédéric , Ribadeau Dumas Antoine , Leriche Laurence , Valent Peter , Arock Michel
Published: April 2003

Mastocytosis is a heterogeneous group of hematopoietic disorders characterized by abnormal growth and accumulation of mast cells (MC) in one or more organs. Clinical symptoms occur as a result of the release of chemical mediators and/or of pathologic infiltration of MC in various tissues. Although the initial events leading to mastocytosis have not yet been unraveled, acquired alterations in the c-kit gene coding for the receptor of stem cell factor (SCF), a major cytokine involved in MC growth, have been described in a significant number of patients. Of particular interest are point mutations resulting in a constitutively activated SCF receptor. Such mutations are probably involved in the abnormal (SCF-independent) proliferation of MC in these patients. New therapeutic strategies may be envisaged to inhibit the deregulated kinase activity of these mutant forms of c-kit.
Histopathological and Immunohistochemical Aspects of Mastocytosis
Authors: Horny Hans-Peter , Valent Peter
Published: April 2003

The diagnosis of mastocytosis is based on histological evidence of a focal increase in tissue mast cells. Immunohistochemical staining with antitryptase antibodies is strongly recommended in all cases of suspected mastocytosis because mast cell infiltrates may be small and scanty. Mastocytosis may be difficult to distinguish from other hematological malignancies, in which an increase in mast cells is frequently seen. The expression of the T cell-associated antigen CD2 has been shown to be exclusively found on neoplastic mast cells in mastocytosis. The demonstration of expression of vascular endothelial growth factor by mast cells is consistent with the finding of angiogenesis which is commonly seen in tissue infiltrates of mastocytosis.
Mast Cell Hyperplasia: Role of Cytokines
Authors: Bischoff Stephan C. , Sellge Gernot
Published: April 2003

Mast cell hyperplasia is found in different pathologies such as chronic inflammatory processes, fibrotic disorders, wound healing or neoplastic tissue transformation. The functional significance of the accumulation of mast cells in these processes is largely unknown. It is now established that bone marrow-derived mast cell progenitors circulate in peripheral blood and subsequently migrate into the tissue where they undergo final maturation under the influence of local microenvironmental factors. Cytokines are of particular importance for mast cell recruitment, development, and function. Stem cell factor (SCF) is a unique mast cell growth factor, since mast cells disappear completely in the absence of SCF. However, several other cytokines such as IL-3 and IL-4 have been shown to influence mast cell proliferation and function also. This review focuses on the role of cytokines in the regulation of mast cell hyperplasia.
Abnormal Expression of CD Antigens in Mastocytosis
Authors: Escribano Luis , Díaz-Agustín Beatriz , Núñez Rosa , Prados Aranzazu , Rodríguez Ramón , Orfao Alberto
Published: April 2003

Human mast cells are myeloid cells derived from human pluripotential CD34+ stem cells. Normal mast cells exhibit a myeloid immunophenotype characterized by the expression of CD117, CD33 and FcΕRI in the absence of reactivity for CD14, CD15 and lymphoid-lineage-associated antigens. Multiparametric flow-cytometric studies have shown that mast cells from mastocytosis display unique immunophenotypic characteristics, including coexpression of CD2 and CD25 antigens together with abnormally high levels of the activation-related antigens CD35, CD63 and CD69 among others. Such aberrant immunophenotypic features are of great relevance in the diagnosis and differential diagnosis of the disease, flow-cytometric immunophenotyping of mast cells representing the most sensitive method for the diagnosis of tissue involvement in mastocytosis. From the pathogenetic point of view, the immunophenotypical patterns described suggest the existence of profound changes regarding the adhesion and activation status of mast cells in mastocytosis and may represent a useful tool for a better understanding of some pathophysiological aspects of the disease.
Spectrum of Associated Clonal Hematologic Non-Mast Cell Lineage Disorders Occurring in Patients with Systemic Mastocytosis
Authors: Sperr Wolfgang R. , Horny Hans-Peter , Valent Peter
Published: April 2003

Systemic mastocytosis (SM) is a myeloproliferative disease affecting multipotent and/or mast cell-committed hematopoietic progenitor cells. In a significant subgroup of patients (10–35%), an associated clonal hematologic non-mast cell lineage disorder (AHNMD) occurs. These AHNMDs can be classified according to recently established WHO criteria. Most AHNMDs resemble myeloid malignancies such as acute myeloid leukemia, myeloproliferative disorders or myelodysplastic syndromes. In only a few cases, lymphoproliferative disorders are diagnosed. Patients with SM-AHNMD have a less favorable prognosis concerning survival when compared to indolent SM. No general guidelines for the treatment of patients with SM-AHNMD have been established so far. A reasonable straightforward approach may be to treat the AHNMD in those patients in the same way as if no coexisting SM exists.
Annexin 5 Messenger Ribonucleic Acid Expression in Pituitary Gonadotropes Is Induced by Gonadotropin-Releasing Hormone (GnRH) and Modulates GnRH Stimulation of Gonadotropin Release
Authors: Kawaminami Mitsumori , Etoh Seishi , Miyaoka Hiroko , Sakai Mina , Nishida Mayumi , Kurusu Shiro , Hashimoto Inoru
Published: April 2003

Our previous studies on annexin 5, a member of the annexin family of proteins, have shown its expression in the anterior pituitary gland, its preferential distribution in gonadotropes, and its increase after ovariectomy. In the present study, we examined (1) whether annexin 5 is synthesized in gonadotropes, (2) whether its expression is under the control of gonadotropin-releasing hormone (GnRH), and (3) the effect of annexin 5 on gonadotropin release. Large cells, also called castration cells, appeared in anterior pituitary tissue 3 weeks after ovariectomy. These cells have been confirmed to be hyperfunctioning gonadotropes and are easily discriminated from other pituitary cells without immunostaining. Using in situ hybridization with a digoxigenin-labeled ribonucleic acid probe, enhanced expression of annexin 5 messenger ribonucleic acid (mRNA) in these gonadotropes was clearly demonstrated. Northern blot analysis showed an increase in the level of annexin 5 mRNA expression 3 weeks after ovariectomy. It was lessened 3 h after the injection of Cetrorelix (GnRH antagonist, 10 µg i.v.). Administration of a GnRH analog [GnRHa; Des-Gly 10 (Pro9) GnRH ethylamide, 0.2 ml of 2.5 µg/ml saline ten times intraperitoneally at 30-min intervals] significantly increased pituitary annexin 5 mRNA. In primary cultures of anterior pituitary cells, recombinant rat annexin 5 stimulated luteinizing hormone (LH) and follicle-stimulating hormone (FSH) release in a dose-dependent manner. Concomitant administration of annexin 5 (1 µg/ml) and GnRHa augmented the LH and FSH release induced by GnRHa. After a 1-hour incubation, cycloheximide (10 µg/ml) apparently inhibited the LH response to GnRHa, while annexin 5 (2 µg/ml) moderated this inhibition. Further, the antisense oligodeoxynucleotide to annexin 5 mRNA blunted the LH response to GnRHa. It is thus concluded that annexin 5 is synthesized in the gonadotropes under the effect of GnRH, and it is suggested that annexin 5 synthesis mediates at least partly GnRH receptor signaling to stimulate gonadotropin secretion.
Weekly Administration of Gemcitabine Plus Docetaxel in Patients with Advanced Breast Cancer: A Phase 1 Study
Authors: Brugnatelli Silvia , Danova Marco , De Bella Manuela Tamburo , Vaglica Marina , Manuguerra Giovanna , Riccardi Alberto , Palmeri Sergio
Published: April 2003

Objective: This study was designed to determine the maximum tolerable dose (MTD) of gemcitabine plus docetaxel, both given on a weekly schedule, in patients with pretreated metastatic breast cancer (MBC). Methods: Heavily pretreated patients with MBC, aged 18–75 years with World Health Organization performance status of 0–2 were enrolled. Three escalating weekly doses of docetaxel (30, 35 and 40 mg/m2) followed by a weekly fixed dose of gemcitabine, 800 mg/m2, were administered on days 1, 8 and 15 of a 28-day cycle. Dose-limiting toxicity (DLT) included grade >3 hematologic toxicity and grade >2 stomatitis, asthenia, diarrhea or organ-specific toxicity (except alopecia). Dose escalation was stopped if ≧3 of 5 patients at any dose level experienced DLT. Results: Eighteen patients (median age 56 years) received a mean of 4.1 (range 1–6) cycles. Asthenia, stomatitis and leukopenia were the main DLTs. One of 5 patients had DLT at dose level 1 and 2 of 5 patients at dose level 2. At dose level 3, 3 of 5 patients had DLTs. Three additional patients treated at dose level 3 confirmed that the MTD had been reached. Therefore, the recommended docetaxel dose in combination with gemcitabine 800 mg/m2 for phase II studies was established at the next lower dose, 35 mg/m2. Of 12 evaluable patients, 7 (58%) achieved an objective response. Conclusions: Gemcitabine 800 mg/m2 plus docetaxel 35 mg/m2 on days 1, 8 and 15 of a 28-day cycle is a safe regimen which shows activity in heavily pretreated patients with MBC. Further phase II investigations with this combination are now warranted.
Correlation between Early Treatment Failure and Ki67 Antigen Expression in Blast Cells of Children with Acute Lymphoblastic Leukaemia before Commencing Treatment
Authors: Nowicki Michał , Miśkowiak Bogdan , Kaczmarek-Kanold Małgorzata
Published: April 2003

Objectives: An attempt has been made to demonstrate the value of the immunocytochemical assay of Ki67 antigen expression in blast cells in children with acute lymphoblastic leukaemia (ALL) before initiation of treatment and its correlation with early treatment failure. Methods: Bone marrow specimens were obtained before treatment from children hospitalised in the years 1997–2000. A total of 60 children diagnosed with ALL have been examined. Immunocytochemical staining for Ki67 expression was based on the ABC technique. Results: Out of 45 children assigned to the low risk group, the presence of Ki67 Ag was demonstrated in 31 cases (68.8%). Out of 15 patients in the high risk group, Ki67 Ag expression in blast cells was positive in 8 children (53.3%). The fraction of immunopositive cells in these groups ranged from 19.8 to 81.3% compared to 5% in the control group. Early treatment failure was observed in both groups and these were closely related to the lack of Ki67 expression observed at the beginning of treatment. Conclusion: The results indicate a possible connection between the Ki67 immunonegative blast pattern and early leukaemia progression. It may also justify routine determination of Ki67 Ag before the treatment of ALL is initiated.
Lymphocyte Activation with Localized pGL1-TNF-Alpha Gene Therapy in a Glioma Model
Authors: Gridley Daila S. , Li Jun , Kajioka Eric H. , Dutta-Roy Radha , Andres Melba L. , Timiryasova Tatyana M. , Fodor Istvan
Published: April 2003

The major goal of this study was to evaluate the effects of tumor necrosis factor-α (TNF-α), delivered as pGL1-TNF-α, on hematological variables, as well as C6 tumor growth in athymic mice treated with and without radiation. pGL1-TNF-α was administered intratumorally at low to high doses (15, 150 and 450 µg) in all three phases of this study. In phase A, pGL1-TNF-α expression within tumors was dose dependent and transient, with highest levels seen at 18 h after injection, whereas no TNF-α protein was detected in plasma. Low erythrocyte counts, hemoglobin, and hematocrit were associated with tumor presence, but the reduction in these variables was most striking in the group receiving 450 µg of pGL1-TNF-α, the group that also exhibited thrombocytopenia at 72 h. In phase B, treatment with pGL1-TNF-α at 15 or 150 µg resulted in the greatest degree of splenomegaly, increased spontaneous blastogenesis by splenocytes, and high leukocyte and lymphocyte numbers in the spleen. In these same two groups, flow cytometry analyses of spleen cells showed that high levels of natural killer (panNK+) cells, B (CD19+) lymphocytes, and cells expressing the CD71 and CD25 activation markers were present (p < 0.05). An enhancing effect was also noted in some of the measurements with parental plasmid p WS4 and tumor presence. In phase C, the slowest tumor progression was observed in the groups receiving 15 and 150 µg pGL1-TNF-α together with radiation; tumor volumes were 51 and 43% smaller, respectively, than for PBS-injected controls by the end of the study. Collectively, these results show that localized treatment with pGL1-TNF-α is hematologically nontoxic at low doses and support the premise that activation of lymphocytes may contribute to the antitumor effects of radiation against a highly aggressive brain tumor.
Anticarcinogenic Effects of Isoflavones May Be Mediated by Genistein in Mouse Mammary Tumor Virus-Induced Breast Cancer
Authors: Mizunuma Hirobumi , Kanazawa Kyotaro , Ogura Shigeto , Otsuka Shin , Nagai Hideo
Published: April 2003

Isoflavones are known to exert anticancer effects. These effects were examined using two isoflavones, biochanin A and daidzein, in a mouse mammary tumor virus (MMTV)-induced spontaneous breast cancer model. Emphasis was placed on isoflavone metabolism by the intestinal microflora and changes in estrogen levels. Germ-free (Gf) mice and their conventionalized (Cv) counterparts were assigned to three diet groups: (1) control diet, (2) biochanin A and (3) daidzein. In all groups, urine was collected from virgin female mice to analyze isoflavone metabolism by high performance liquid chromatography. These studies revealed changes of biochanin A into genistein, and of daidzein into equol, which were accelerated in the Cv animals. However, the Gf mice could not transform biochanin A into genistein, or daidzein into equol. Estrogen levels in the control and daidzein diet groups were lower in the Gf mice than in the Cv mice. The biochanin A group showed no differences in estrogen levels between the Cv and Gf animals. Four-week-old male and female animals were paired in the Gf and Cv groups. The female animals delivered and lactated repeatedly and were observed for the development of mammary cancer by palpation, twice weekly, until 15 months of age. The Cv mice showed a significantly lower incidence of breast cancer in the biochanin A diet group than in the control or daidzein groups (p < 0.05). These results suggest that the anticarcinogenic effects in this system might be produced not by daidzein or equol, but by biochanin A and/or genistein. In the Gf animals, the incidence of breast cancer was significantly higher in the biochanin A group than in the control group (p < 0.05), probably due to the increased level of estradiol in the former group. The biochanin A group tended to have a higher incidence of breast cancer than the daidzein group in the Gf group, although no significant differences were noted. Thus, no anticarcinogenic effect was produced by biochanin A alone in the Gf mice. In view of the results presented, genistein derived from biochanin A following metabolic processes in the intestinal microflora most likely acts as an inhibitor in breast carcinogenesis; biochanin A is most likely a precursor of genistein.
Are the Effects of Tamoxifen on the Serum Lipid Profile Modified by Apolipoprotein E Phenotypes?
Authors: Liberopoulos Evagellos , Karabina Sonia-Athena , Tselepis Alexandros , Bairaktari Eleni , Nicolaides Costas , Pavlidis Nikolaos , Elisaf Moses
Published: April 2003

Background: Tamoxifen has favorable effects on the serum lipid profile. It has been suggested that the apolipoprotein (Apo) E phenotype can influence serum lipid parameters; the ApoE allele 4 (ApoE4) is associated with higher total and low-density lipoprotein (LDL) cholesterol levels. The ApoE phenotype also affects lipid responses to diets or treatment with statins. However, the effect of tamoxifen on the lipid profile in different ApoE phenotypes is unknown. Patients and Methods: In the present study, we evaluated the effects of tamoxifen on the serum lipid profile in 11 ApoE4-positive postmenopausal women with breast cancer (phenotypes 3/4 and 4/4) compared with 33 ApoE4-negative women (phenotypes 3/2 and 3/3). Serum lipid parameters [high-density (HDL), LDL and total cholesterol, triglycerides, ApoAI, ApoB and lipoprotein (a)] were measured after an overnight fast before treatment and after 3 and 12 months. ApoE isoforms were determined by isoelectric focusing of delipidated very-low-density lipoproteins (VLDL). Results: During the follow-up period, serum levels of total and LDL cholesterol and ApoB decreased significantly in both groups, but no significant differences were found. Concentrations of serum HDL cholesterol were not significantly different between both groups. However, serum ApoAI levels increased significantly in ApoE4-negative subjects (p = 0.00005), but no significant changes in ApoE4-positive women were observed. Serum triglyceride levels increased by 23.2% (p < 0.05) in ApoE4-positive patients, but they did not change significantly in ApoE4-negative patients. The LDL/HDL cholesterol ratio decreased similarly in the two groups, but the ApoAI/ApoB ratio, which may be a better predictor of cardiovascular events, significantly changed in the ApoE4-negative subjects. Finally, the median level of Lp(a) decreased by 43.4% in the ApoE4-negative patients, whereas it did not change significantly in the ApoE4-positive group. Conclusion: In postmenopausal Greek women with breast cancer, the levels of Lp(a) and triglycerides and the ApoAI/ApoB ratio respond more favorably to tamoxifen treatment in ApoE4-negative than in ApoE4-positive patients.
Ultrasound Biomicroscopy of Residual Vitreous Base after Vitreoretinal Surgery
Authors: Pierro Luisa , Azzolini Claudio , Codenotti Marco , Locatelli Cristina , Tremolada Gemma , Brancato Rosario
Published: April 2003

Residual vitreous base after vitreoretinal surgery was evaluated by ultrasound biomicroscopy (UBM). Twenty aphakic and pseudoaphakic patients (20 eyes) undergoing surgery for different vitreoretinal diseases were evaluated by high-frequency (50 MHz), high-resolution (50 µm) UBM, performed the day before surgery, weekly up to 1 month after surgery and then monthly. One week after surgery, the vitreous remnants were ‘hardly visible’ in 3 cases, ‘visible’ in 6 cases and ‘highly visible’ in 11. At the end of the follow-up (2.2 months), the ‘hardly visible’ cases increased to 6 and the ‘visible’ cases to 12, while the ‘highly visible’ cases decreased to 2. UBM demonstrated that vitreous base remnants were present in spite of accurate surgery; a spontaneous volume reduction of vitreous was observed during the follow-up.
Farbduplexsonographie der Temporalarterien: Biopsieersatz bei Arteriitis temporalis?
Authors: Schmid Rudenz , Hermann Marianne , Yannar Ahmet , Baumgartner Ralf W.
Published: April 2003

The diagnosis of temporal arteritis (TA) is generally confirmed by biopsy. To investigate the diagnostic accuracy of color duplex sonography (CDS), both temporal arteries of 20 patients with suspected TA were prospectively insonated prior to biopsy. Detection of ≧1 hypoechogenic perivascular halo was used as CDS criterion, a temporal artery biopsy and the criteria of the American College of Rheumatology (ACR) as references. The frequency of halo disappearance after 3 months of steroid therapy was also studied. CDS showed TA in 6, biopsy in 12 and ACR criteria in 15 patients. CDS sensitivity was 50 and 40%, and specificity 100%, using the biopsy and the ACR criteria, respectively. After 3 months of steroid treatment, 1 patient still showed halos. In conclusion, detection of halos confirms, whereas the absence of halos does not exclude the diagnosis of TA suggesting that ultrasound may replace biopsy in single patients with typical clinical signs and symptoms and a halo.
Postural Modifications of the Oscillatory Potentials of the Electroretinogram in Primary Open-Angle Glaucoma
Authors: Ferreri Giuseppe , Buceti Rosaria , Ferreri Felicia M.B. , Roszkowska Anna M.
Published: April 2003

The postural variations in the retinal microcirculation in glaucomatous patients were studied by evaluation of the oscillatory potentials (OPs) of the ERG. The OPs in scotopic adaptation were examined in subjects with primary open-angle glaucoma and in an age- and sex-matched normal control group in different body positions (seated, supine, anti-Trendelenburg, Trendelenburg). In the seated position, the difference of mean OP amplitude between the control group and the glaucomatous patients was highly significant (p < 0.001). In the normal subjects the OP amplitude in the Trendelenburg position was statistically lower with respect to the values obtained in all other positions (p < 0.05). In the glaucomatous patient group, the OP amplitude in the anti-Trendelenburg position was increased as compared to the other positions (p < 0.05). The study showed a reduction in amplitude of OPs in glaucomatous patients and their different behaviour in both groups with changes in body position.
Effect of Botulinum Toxin A on Facial Wrinkle Lines in Koreans
Authors: Lew Helen , Yun Young Soo , Lee Sang Yeul , Kim Sung Joo
Published: April 2003

Two kinds of botulinum toxin type A were clinically evaluated in rhytidectomy. Twenty Korean patients with facial wrinkles were fully assessed following treatments with random injections. The mean degree of wrinkles before the injections was 2.83 and the mean corrective effect was 70.0% at least 3 months afterward. The effect lasted less than 6 months in only 9 cases. The complications were tingling sensations in 3 cases (15.0%), temporary lid swelling in 5 cases (25.0%) and lagophthalmos in 3 cases (15.0%). No serious or permanent adverse effects were observed. Botulinum toxin type A rhytidectomy was a very effective method of removing various facial wrinkles although the treatment for complications and side effects will need to be considered.
Ocular Firework Injuries at New Year’s Eve
Authors: Sacu Stefan , Ségur-Eltz Nikolaus , Stenng Karin , Zehetmayer Martin
Published: April 2003

Purpose: To prospectively study mechanisms and injury characteristics of ocular firework burns. Methods: A prospective analysis of all patients with firework injuries attending the Department of Ophthalmology, University of Vienna, between 1994 and 2001. We looked for classes of fireworks and mechanisms of injuries. The number of diagnoses was established and their severity classified. Results: During this period (8 years) we identified 116 eyes of 102 patients. 67/102 (66%) of all injuries were caused by class II and III fireworks. Patients ranged between 4 and 83 years. Ocular injuries occurred more frequently in males (69, 68%) and affected the right eye in 53 patients (52.5%). Ocular firework injuries to minors (under the age of 18 years) occurred in 48 (49%). The most common types of injury were skin and corneal erosions and abrasions (32/116, 28%). 11/116 eyes (10%) had severe injuries. 8 of 11 severe injuries (72%) occurred in minors (<18 years). 2 patients (1.7%) developed permanent blindness. In 32/116 patients (28%), the ocular trauma resulted in visual impairment, mainly due to corneal scars or retinal pathologies. In all patients, the severity index was 1.4 ± 0.8. In minors the severity index was 1.6 ± 1.0, in adults 1.2 ± 0.5 (p = 0.013). Conclusion: Injuries in minors were significantly more severe than those in adults. Possible preventive measures include legislation, education of minors and eye protection.
Coagulative, Fibrinolytic and Metabolic Pattern in Patients with Central Retinal Vein Occlusion
Authors: Fruschelli M. , Puccetti L. , Bruni F. , Auteri A.
Published: April 2003

Central retinal vein occlusion (CRVO) is an important cause of visual loss. Many risk factors have been associated with CRVO onset at various ages. Among them diabetes mellitus, hypertension, immunologic disorders, increase in blood viscosity and coagulation, decrease of fibrinolysis have been reported in many subjects. The aim of our study was to detect the metabolic, coagulative and fibrinolytic pattern in 54 patients (26 men, 28 women, mean age 50.4 ± 12.3) affected by CRVO. We excluded from the study patients with other ocular disorders. A fibrinolytic impairment is the most common feature in our population. It occurs either in dysmetabolic or in nonmetabolic subjects. Such data suggest a prominent role of the fibrinolytic system in the pathogenesis of CRVO.
The Effects of Bupivacaine and Lidocaine on the Corneal Endothelium when Applied into the Anterior Chamber at the Concentrations Supplied Commercially
Authors: Guzey Mustafa , Satici Ahmet , Dogan Zeki , Karadede Sezin
Published: April 2003

Purpose: To investigate the direct toxic effects of bupivacaine HCl 0.5% and lidocaine HCl 2%, two commonly used injectable local anesthetic agents, on the corneal endothelium when applied intracamerally. Methods: Two groups were formed, each consisting of 12 pigmented rabbits, and 0.2 ml of the anesthetic agent were injected intracamerally into the right eyes. The central corneal thicknesses and corneal clarities were evaluated preoperatively and at 3, 6, 9, 12 h and 1, 3, 7 days postoperatively. While the central corneal thicknesses were evaluated by ultrasonic pachymetry, the corneal opacification scored between 0 and 3 was assessed by biomicroscopic examination and photographs. Results: Both bupivacaine and lidocaine caused corneal thickening in the 3- to 12-hour measurements. In addition, there was significant corneal opacification in both groups in the 3-hour and 3-day measurements. The corneal thickening and corneal opacification determined during 3- and 6-hour measurements in the eyes which received intracameral bupivacaine were significantly higher than those determined in the lidocaine-injected group. In both groups, the corneal thickness and corneal clarity scores returned to the preoperative values on the 1st and 7th days, respectively. Conclusions: When applied into the anterior chamber at the concentrations supplied commercially, both bupivacaine and lidocaine cause statistically significant corneal thickening and clinically significant corneal opacification. It should be noted that the injection of these agents into the anterior chamber during the operation at the concentrations supplied commercially may be a potential risk factor for endothelial injury.