bluuebunny/crossref_metadata_2025 huggingface dataset:
The Clinical Safety of Viagra
Authors: LIM PETER H.C. , MOORTHY P. , BENTON KENNETH G.F.
Published: July 2009

Abstract: Erectile dysfunction (ED) in men is amenable to correction with Viagra in a majority of patients. The accumulated experience of prescribing Viagra across the broad continuum of men suffering from ED is sufficient for a meaningful assessment of the safety of Viagra in clinical practice. The use of Viagra necessitates caution in cardiac failure and when used within six months of acute myocardial infarction and stroke. It is inadvisable in patients with unstable angina pectoris. The co‐administration of Viagra with organic nitrates, for example, glyceryl trinitrate or isosorbide dinitrate, is unsafe. The relative contraindications to Viagra in cardiovascular disease are uncontrolled hypertension and impaired cardiac reserve. With respect to interactions with other drugs, the potential influence on the metabolism of Viagra by medications that affect the cytochrome‐P‐450 system does not translate into clinical effects. The vasodilatory properties of sildenafil citrate are largely responsible for unwanted effects. The most common side effects are headache, flushing (due to vasodilation), and dyspepsia (due to relaxation of the smooth muscle of the gastroesophageal sphincter with reflux). In the recommended single‐dose range (25‐100 mg), the use of Viagra for erectile dysfunction, in the absence of contraindications, is extremely safe provided the drug is taken under proper conditions.
Glucocorticoid Receptor Downregulation in Early Diagnosed Rheumatoid Arthritis
Authors: HUISMAN A. MARGRIET , VAN EVERDINGEN A. AMALIA , WENTING MARION J.G. , VAN REESEMA DICK R. SIEWERTSZ , LAFEBER FLORIS P.J.G. , JACOBS JOHANNES W.G. , BIJLSMA JOHANNES W.J.
Published: July 2009

Abstract: In patients with rheumatoid arthritis (RA) with a mean duration of six years, glucocorticoid receptor (GR) downregulation has been reported without change in cortisol levels. This phenomenon might play a role in the etiopathogenesis of RA. If this is the case, GR downregulation should also be found in early‐diagnosed RA. We compared GR expression and cortisol levels of early RA patients with those of sex and age matched healthy controls. In early (disease duration less than one year) RA patients (52F/29M; mean [SE] age 63 ± 2 years) and in 39 controls (23F/16M; mean age 63 ± 2 years) blood samples were taken between 8:00 and 10:00 a.m. GR expression (GR number), cortisol levels (fluorescence polarization immunoassay, FPIA), ESR, CRP, and the presence of swollen and painful knees were determined. RA was diagnosed according to the ACR classification criteria. In early RA a significantly lower GR expression was found compared to controls. Interestingly, cortisol levels were also significantly lower. Both findings were due to the observed differences in the female population. In the male population no differences were found. No correlations were found among parameters of disease activity, nor was there a relation between GR expression and serum cortisol levels. In female patients with early‐diagnosed RA a decrease in glucocorticoid receptor expression was found as well as a decrease in cortisol levels. Since normal glucocorticoid receptor expression was found in the male population with early‐diagnosed RA, it remains questionable if glucocorticoid receptor expression is causally involved in pathogenesis of RA.
Androgens and Estrogens Modulate the Immune and Inflammatory Responses in Rheumatoid Arthritis
Authors: CUTOLO MAURIZIO , SERIOLO BRUNO , VILLAGGIO BARBARA , PIZZORNI CARMEN , CRAVIOTTO CHIARA , SULLI ALBERTO
Published: July 2009

Abstract: Generally, androgens exert suppressive effects on both humoral and cellular immune responses and seem to represent natural anti‐inflammatory hormones; in contrast, estrogens exert immunoenhancing activities, at least on humoral immune response. Low levels of gonadal androgens (testosterone/dihydrotestosterone) and adrenal androgens (dehydroepiandrosterone and its sulfate), as well as lower androgen/estrogen ratios, have been detected in body fluids (that is, blood, synovial fluid, smears, salivary) of both male and female rheumatoid arthritis patients, supporting the possibility of a pathogenic role for the decreased levels of the immune‐suppressive androgens. Several physiological, pathological, and therapeutic conditions may change the sex hormone milieu and/or peripheral conversion, including the menstrual cycle, pregnancy, the postpartum period, menopause, chronic stress, and inflammatory cytokines, as well as use of corticosteroids, oral contraceptives, and steroid hormonal replacements, inducing altered androgen/estrogen ratios and related effects. Therefore, sex hormone balance is still a crucial factor in the regulation of immune and inflammatory responses, and the therapeutical modulation of this balance should represent part of advanced biological treatments for rheumatoid arthritis and other autoimmune rheumatic diseases.
Melatonin Participates in the Control of Testosterone Secretion from Rat Testis
Authors: VALENTI SANDRA , GIUSTI MASSIMO
Published: July 2009

Abstract: The androgen milieu plays an important role in the control of immunity, and melatonin (MLT) is known to modulate the immune response as well. Our recent studies have focused on the role of MLT in controlling androgen secretion from rat testis. The presence of MLT receptors, their modulation, and the effect of MLT on steroidogenesis have been studied on Percoll‐purified rat Leydig cells (LCs) cultured in vitro. MLT receptors present in adult rat LCs (Bmax= 46.7 ± 3.5 fmol/mg protein; Kd= 88.7 ± 6.2 pmol/l) are coupled through a pertussis toxin‐sensitive G‐protein and are downregulated by prolonged exposure to MLT itself. When acutely added for three hours, MLT (0.4‐400 nM) inhibits T secretion in response to LH, forskolin, and GnRH, but sensitizes cAMP‐dependent T secretion when present in the preincubation media (16 h). The acute inhibition of adenylyl cyclase and the block of 17‐20‐desmolase (with increased 17‐OH‐progesterone) and of calcium release from the intracellular stores, followed by sensitization of the adenylyl cyclase activity in the long term, are described as mechanisms of MLT action. Ultimately, our studies suggest that MLT is likely to control the immune response not only through its direct effects, but also by finely modulating the androgen milieu.
Fatigue in Daily Life in Patients with Primary Sjögren's Syndrome and Systemic Lupus Erythematosus
Authors: GODAERT G. L.R. , HARTKAMP A. , GEENEN R. , GARSSEN A. , KRUIZE A. A. , BIJLSMA J. W.J. , DERKSEN R.H. W.M.
Published: July 2009

Abstract: Fatigue is a well‐recognized complaint with major impact on daily life in primary Sjögren's syndrome (PSS) and systemic lupus erythematosus (SLE). Previous research has not taken into account several crucial aspects of fatigue. This study examined various aspects of fatigue in the daily life of patients with PSS and SLE and in healthy controls. We compared age‐adjusted, repeated measurements of fatigue across the day of female patients with SLE (n= 20, mean age 43.4 ± 11.3), with PSS (n= 28, mean age 53.7 ± 13.9) and healthy participants (n= 30, mean age 50.5 ± 13.4). General and physical fatigue was significantly higher in patients than in healthy participants. Groups did not differ with respect to average levels of reduced motivation or mental fatigue. Both general and physical fatigue and reduced activity varied significantly during the day. Adjusting for depressive symptoms, groups showed significantly different time courses during the day. In healthy participants and patients with SLE, fatigue first decreased and then increased, whereas a rather opposite course—at least for the first part of the day—was observed in patients with PSS. Using an ecologically valid assessment method, we demonstrated substantially higher levels of daily fatigue in SLE and PSS patients as compared to healthy participants, thereby jeopardizing these patients' quality of life. The effect of disease on variations in fatigue over the day should be the subject of further inquiry, especially as it might clarify underlying mechanisms.
Blunted ACTH and Cortisol Responses to Systemic Injection of Corticotropin‐Releasing Hormone (CRH) in Fibromyalgia
Authors: RIEDEL WALTER , SCHLAPP ULRIKE , LECK STEFANIE , NETTER PETRA , NEECK GUNTHER
Published: July 2009

Abstract: Thirteen female patients suffering from fibromyalgia (FM) and thirteen female age‐matched controls were intravenously injected with a bolus dose of 100 μg corticotropin‐releasing hormone (CRH), and the evoked secretion pattern of ACTH, cortisol, somatostatin, and growth hormone (GH) was followed up for two hours, together with the plasma levels of CRH. The increases of ACTH and cortisol following CRH were not significantly different between controls and FM patients. The increase of plasma CRH following its injection was significantly higher in FM patients and lasted about 45 min, paralleled by an increase of somatostatin with a similar time course. Basal GH levels were significantly lower in FM patients. GH increased in FM patients 90 min after injection of CRH, coincident with decreasing CRH and somatostatin levels, while GH levels in controls rather decreased with the lowest values occurring 90 min after CRH. The results support the concept that the hormonal secretion pattern frequently observed in FM patients is primarily caused by CRH, possibly as a response to chronic pain and stress. The elevated levels of CRH in the circulation of FM patients suggest elevated levels of CRH‐binding protein, which could explain why the levels of ACTH and cortisol between controls and FM following CRH do not differ.
Regulation of Apoptosis in Endocrine Autoimmunity
Authors: SALMASO C. , BAGNASCO M. , PESCE G. , MONTAGNA P. , BRIZZOLARA R. , ALTRINETTI V. , RICHIUSA P. , GALLUZZO A. , GIORDANO C.
Published: July 2009

Abstract: Dysregulation of apoptosis is associated with the pathogenesis of organ‐specific autoimmune diseases, through altered target organ susceptibility. Apoptosis signaling pathways can be initiated through activation of death receptors such as Fas. A comparative analysis of the expression of Fas and FasL, the antiapoptotic molecule Bcl‐2, and apoptosis in both thyrocytes and thyroid‐infiltrating lymphocytes (TILs) from patients with either Graves' disease (GD) or Hashimoto's thyroiditis (HT) was performed. GD thyrocytes expressed less Fas than HT thyrocytes, whereas GD TILs had higher levels of Fas and FasL than HT TILs. GD thyrocytes expressed higher levels of Bcl‐2 compared with HT thyrocytes. The opposite pattern was observed in GD (low Bcl‐2) and HT (high Bcl‐2) TILs. Consistently, thyrocyte apoptosis was marked in HT and poor in GD thyroids, and TIL apoptosis was marked in GD and poor in HT. Our findings suggest that in GD thyroid the regulation of Fas/FasL/Bcl‐2 favors apoptosis of infiltrating lymphocytes. Moreover, the reduced levels of Fas/FasL and increased levels of Bcl‐2 should favor thyrocyte survival and hypertrophy associated with stimulatory thyroid‐stimulating hormone receptor antibodies. In contrast, the regulation of Fas/FasL/Bcl‐2 expression in HT can promote thyrocyte apoptosis via homophylic Fas‐FasL interactions, and a gradual reduction in thyrocyte numbers leading to hypothyroidism. Fas‐mediated apoptosis may be a general mechanism of cell damage in destructive organ‐specific autoimmunity.
Role of the PKA‐Regulated Transcription Factor CREB in Development and Tumorigenesis of Endocrine Tissues
Authors: ROSENBERG D. , GROUSSIN L. , JULLIAN E. , PERLEMOINE K. , BERTAGNA X. , BERTHERAT J.
Published: July 2009

Abstract: The cAMP pathway plays a major role in the development of endocrine tissues and various molecular defects of key components of this pathway (G protein, receptors, PKA, etc.) have been observed in endocrine tumors. The ubiquitous transcription factor CREB (cAMP‐response element binding protein) binds to the cAMP response element (CRE) and stimulates transcription after phosphorylation on Ser133 by PKA. The CREB family of transcription factors contains three members: CREB, CREM, and ATF‐1. Targeted expression of dominant‐negative mutants of CREB in transgenic mice leads to somatotrophs or thyroid hypoplasia. GH‐secreting adenomas are benign secreting tumors expressing an activated mutant Gαs protein (Gsp) in about 40% of cases. In GH‐secreting adenomas CREB is always expressed and often highly phosphorylated. The CREM isoform ICER is stimulated by cAMP, and its expression is increased in Gsp‐harboring tumors. After transfection in pituitary somatotroph cells, activating mutations of Gs protein (Gsp) and overexpression of wild‐type GαS stimulate transcription of various CRE‐containing promoters via CREB in a Ser133‐specific‐dependent manner. Activation of the cAMP pathway by ACTH is required for adrenal cortex (AdCx) maintenance and steroidogenesis. CREB is expressed in normal AdCx. Alterations of CRE binding proteins with loss of CREB expression and compensatory overexpression of CREMτ is observed in the human adrenocortical cancer cell line H295R. Similar alterations are found at the protein level in human malignant adrenocortical tumors. In conclusion, the CREB family of transcription factors plays an important role in the development, differentiation, and proliferation of endocrine tissues. Various alterations of the CREB family of transciption factors can be observed in endocrine tumors.
The Essential Role of RIα in the Maintenance of Regulated PKA Activity
Authors: AMIEUX PAUL S. , MCKNIGHT G. STANLEY
Published: July 2009

Abstract:Cloning of the individual regulatory (R) and catalytic (C) subunits of the cAMP‐dependent protein kinase (PKA) and expression of these subunits in cell culture have provided mechanistic answers about the rules for PKA holoenzyme assembly. One of the central findings of these studies is the essential role of the RIα regulatory subunit in maintaining the catalytic subunit under cAMP control. The role of RIα as the key compensatory regulatory subunit in this enzyme family was confirmed by gene knockouts of the three other regulatory subunits in mice. In each case, RIα has demonstrated the capacity for significant compensatory regulation of PKA activity in tissues where the other regulatory subunits are expressed, including brain, brown and white adipose tissue, skeletal muscle, and sperm. The essential requirement of the RIα regulatory subunit in maintaining cAMP control of PKA activity was further corroborated by the knockout of RIα in mice, which results in early embryonic lethality due to failed cardiac morphogenesis. Closer examination of RIα knockout embryos at even earlier stages of development revealed profound deficits in the morphogenesis of the mesodermal embryonic germ layer, which gives rise to essential structures including the embryonic heart tube. Failure of the mesodermal germ layer in RIα knockout embryos can be rescued by crossing RIα knockout mice to Cα knockout mice, supporting the conclusion that inappropriately regulated PKA catalytic subunit activity is responsible for the phenotype. Isolation of primary embryonic fibroblasts from RIα knockout embryos reveals profound alterations in the actin‐based cytoskeleton, which may account for the failure in mesoderm morphogenesis at gastrulation.
Gsα Mutations and Imprinting Defects in Human Disease
Authors: WEINSTEIN LEE S. , CHEN MIN , LIU JIE
Published: July 2009

Abstract: Gs is the ubiquitously expressed heterotrimeric G protein that couples receptors to the effector enzyme adenylyl cyclase and is required for receptor‐stimulated intracellular cAMP generation. Activated receptors promote the exchange of GTP for GDP on the Gs α‐subunit (Gsα), resulting in Gs activation; an intrinsic GTPase activity of Gsα deactivates Gs by hydrolyzing bound GTP to GDP. Mutations of Gsα residues involved in the GTPase reaction that lead to constitutive activation are present in endocrine tumors, fibrous dysplasia of bone, and McCune‐Albright syndrome. Heterozygous loss‐of‐function mutations lead to Albright hereditary osteodystrophy (AHO), a disease characterized by short stature, obesity, and skeletal defects, and are sometimes associated with progressive osseous heteroplasia. Maternal transmission of Gsα mutations leads to AHO plus resistance to several hormones (e.g., parathyroid hormone) that activate Gs in their target tissues (pseudohypoparathyroidism type IA), while paternal transmission leads only to the AHO phenotype (pseudopseudohypoparathyroidism). Studies in both mice and humans demonstrate that Gsα is imprinted in a tissue‐specific manner, being expressed primarily from the maternal allele in some tissues and biallelically expressed in most other tissues. This likely explains why multihormone resistance occurs only when Gsα mutations are inherited maternally. The Gsα gene GNAS1 has at least four alternative promoters and first exons, leading to the production of alternative gene products including Gsα, XLαs (a novel Gsα isoform expressed only from the paternal allele), and NESP55 (a chromogranin‐like protein expressed only from the maternal allele). The fourth alternative promoter and first exon (exon 1A) located just upstream of the Gsα promoter is normally methylated on the maternal allele and is transcriptionally active on the paternal allele. In patients with parathyroid hormone resistance but without AHO (pseudohypoparathyroidism type IB), the exon 1A promoter region is unmethylated and transcriptionally active on both alleles. This GNAS1 imprinting defect is predicted to decrease Gsα expression in tissues where Gsα is normally imprinted and therefore to lead to renal parathyroid hormone resistance.
A Review of Mutual Transmission of Important Infectious Diseases between Livestock and Wildlife in Europe
Authors: FRÖLICH K. , THIEDE S. , KOZIKOWSKI T. , JAKOB W.
Published: July 2009

Abstract: Oral vaccination of red foxes against rabies has been practiced in Europe since 1978 and has succeeded in greatly reducing the occurrence of this disease in foxes: this is an example of coordinated activity against a disease that affects both wild and domestic animals as well as humans. Some examples of diseases that affect both domestic and wild animals in Europe are: classical swine fever (hog cholera) in wild boars and domestic swine; myxomatosis and rabbit hemorrhagic disease in domestic and wild rabbits; bovine viral diarrhea (BVD) in cattle and roe deer; contagious ecthyma in domestic sheep and goats and also in, e.g., chamois, muskox, and reindeer; Mycobacterium bovis in cattle, wild boars, badgers, and deer; and brucellosis in a broad range of livestock and wildlife in all European countries. In addition, serological surveys performed in different free‐ranging ungulate species revealed the presence of alphaherpesviruses related to bovine herpesvirus‐1 in 7 European countries; and a study of malignant catarrhal fever in deer in Germany might indicate that in this case sheep are the main reservoir species. Although many data on infectious diseases are available in various European countries, there is more need for systematic surveillance and coordinated research.
Health Monitoring and Conservation of Wildlife in Sweden and Northern Europe
Authors: MÖRNER TORSTEN
Published: July 2009

Abstract: Monitoring of wildlife diseases started in Scandinavia as sporadic postmortem examinations in the early 20th century. In 1945 a monitoring program for wildlife health was initiated. The program is today an integrated part of the National Environmental Monitoring programs in Sweden. The total material today comprises more than 80000 recorded investigations. There are similar programs in Denmark, Finland, and Norway, but no comparable program exists in other Northern European countries. The program has led to discoveries of new diseases in many mammals and birds, and demonstration of several environmental pollutants, such as mercury, lead, and cadmium, in wildlife. The success of the program is due to several factors such as: a rich wildlife with economically important game species; location of the program at the National Veterinary Institute with excellent facilities and competent staff, good cooperation with hunter and conservation organizations, stable financial support, a cool climate, and a successful relationship with media. Current work also includes more focused investigations on different specific pathogens and pollutants, as well as investigations in single animal species.
Global Analysis of Brucella melitensis Proteomes
Authors: MUJER CESAR V. , WAGNER MARY ANN , ESCHENBRENNER MICHEL , HORN TROY , KRAYCER JO ANN , REDKAR RAJENDRA , HAGIUS SUE , ELZER PHILIP , DELVECCHIO VITO G.
Published: July 2009

Abstract: Brucella melitensis is a facultative, intracellular, gram‐negative cocco‐bacillus that causes Malta fever in humans and brucellosis in animals. There are at least six species in the genus, and the disease is classified as zoonotic because several species infect humans. Using 2‐D gel electrophoresis and mass spectrometry, we have initiated (i) a comprehensive mapping and identification of all the expressed proteins of B. melitensis virulent strain 16M, and (ii) a comparative study of its proteome with the attentuated vaccinal strain Rev 1. Comprehensive proteome maps of all six Brucella species will be generated in order to obtain vital information for vaccine development, identification of pathogenicity islands, and establishment of host specificity and evolutionary relatedness.
Expression of RoTat 1.2 Cross‐reactive Variable Antigen Type in Trypanosoma evansi and T. equiperdum
Authors: CLAES FILIP , VERLOO D. , DE WAAL D. T. , URAKAWA T. , MAJIWA P. , GODDEERIS B. M. , BÜSCHER P.
Published: July 2009

Abstract: The variable antigen type (VAT) RoTat 1.2 has been cloned from a T. evansi strain, isolated in 1982 from a water buffalo in Indonesia. All T. evansi isolates hitherto tested express this VAT. In a study on the differential diagnosis of T. equiperdum and T. evansi in horses, we investigated serological evidence for the expression of RoTat 1.2 in 11 T. evansi and six T. equiperdum populations originating from Asia, Europe, Africa, and the Americas. Preinfection sera and sera of days 7, 14, 25, and 35 post‐infection (p.i.) were analyzed for the presence of antibodies reactive with RoTat 1.2 in immune trypanolysis, ELISA/T. evansi and CATT/T. evansi. Within the duration of the experiment, all rabbits infected with T. evansi became positive in the three serological tests. Five out of six rabbits infected with T. equiperdum also became positive in the three tests. Only one T. equiperdum strain (the OVI strain from South Africa) did not induce the production of antibodies reactive with RoTat 1.2 and thus might not contain or express a VSG that shares epitopes similar to those on the RoTat 1.2 VSG. The data lead to the conclusion that T. equiperdum can express VSGs containing epitopes serologically similar to those in the T. evansi RoTat 1.2 VAT. This explains, in part, why the antibody detection tests based on Ro Tat 1.2 VSG cannot reliably distinguish between the infections caused by T. evansi and those caused by T. equiperdum. There are no data that contradict the possibility that the putative T. equiperdum strains, which express VSGs with epitopes similar to those on RoTat 1.2, are actually T. evansi.
The Fencing Issue Relative to the Control of Foot‐and‐Mouth Disease
Authors: SUTMOLLER PAUL
Published: July 2009

Abstract: Certain livestock diseases in sub‐Saharan Africa, such as foot‐and‐mouth disease are difficult to control because of the large numbers of infected wildlife hosts. These wildlife disease reservoirs form a continuous hazard of transmittal of the diseases to domestic livestock, which limits the access of livestock products from southern Africa to international markets. The disease reservoirs are often found in border areas between countries with susceptible species and infected reservoir animals continuously crossing the border. A regional approach to disease control is probably the only way to achieve any real progress. Here we review the positive and negative attributes of fencing as a control mechanism for disease transmission.
Avian Cholera on North Coast California
Authors: BOTZLER RICHARD G.
Published: July 2009

Abstract: Between 1945 and 2001, avian cholera (Pasteurella multocida infection) was confirmed at 27 epizootics in 18 different years on northcoastal California. Estimated mortality ranged from 1 to 6750 birds per site, with a median total mortality of about 1000 birds per year. Eight epizootics involved < 150 birds; thus, minor epizootics were common. Annual total wildfowl mortality ranged from 0.4% to 7.0% of estimated live populations; median annual mortality for American coots (Fulica americana) (11.5%) surpassed that of tundra swans (Cygnus columbianus) (0.2%) and ducks (0.2%). Coots comprised > 50% of total wildfowl mortality in 16 of 17 epizootics. Overall, coots comprised 82% of known avian cholera mortality, but only 34% of the live wildfowl present; ducks and swans died much less frequently. Wildfowl at one site consistently died in a sequential pattern; there was no sequential mortality at other sites.
Bovine Tuberculosis in Michigan Wildlife
Authors: PAYEUR JANET B. , CHURCH S. , MOSHER L. , ROBINSON‐DUNN B. , SCHMITT S. , WHIPPLE D.
Published: July 2009

Abstract: White‐tailed deer in Michigan are now recognized as a reservoir host of bovine tuberculosis (TB). It has been determined that the most likely cause of bovine TB infection in the deer is from congregating in artificially high numbers at feed sites. The presence of a wildlife reservoir of TB in Michigan poses a serious threat to the control and eradication programs that are now in their final stages in the United States.
Control and Eradication of Chelonian Tick Infestations, with Particular Reference to Vectors of Heartwater
Authors: BURRIDGE MICHAEL J. , SIMMONS LEIGH‐ANNE , PETER TREVOR F. , MAHAN SUMAN M.
Published: July 2009

Abstract: Studies using the African tortoise tick (Amblyomma marmoreum) and leopard tortoises (Geochelone pardalis) demonstrated that cyfluthrin and permethrin were safe and efficacious acaricides for control of Amblyomma ticks on tortoises. A protocol was developed that successfully eradicated an A. sparsum infestation from a tortoise breeding facility in Florida. It involved treatment of all tortoises with a permethrin formulation, followed by treatment of the premises with a cyfluthrin formulation. Sentinel tortoises were later placed on the treated premises to establish successful tick eradication.
Eradication of the Tropical Bont Tick in the Caribbean
Authors: PEGRAM RUPERT G. , GERSABECK EDWARD F. , WILSON DAVID , HANSEN JORGEN W.
Published: July 2009

Abstract: The progress and problems in the Caribbean Amblyomma Program (CAP) are reviewed since its inception in 1995. During 1998, there were funding and administrative management problems. USDA resolved the acute funding crisis, and after three years of negotiation, the CAP has now secured an additional euro 1.5 million from the European Community. Changes in administration in 1998 included the withdrawal of IICA from the program, and the transition during the decentralization of administrative and financial management from FAO headquarters to the Regional Office for Latin America and the Caribbean, based in Chile. A general overview of technical progress and one case study, St. Kitts, is presented. One major concern that emerged during 2000 is that the elimination of the small remaining tropical bont tick (TBT) “hot spots” in both St. Kitts and St. Lucia remained elusive. Why is this so? Egrets? Alternative residual hosts? Or is it fatigue in both technical and administrative management functions? Of even greater concern is the finding of two, apparently new, foci in St. Croix (USVI) in the north and St. Vincent in the south. A critical overview of the program has identified one major remaining constraint—an appropriate management support function at both regional and, in some countries, at the national level. A proposal for a revised management strategy, coupled with the identification of a future strategy to succeed the CAP, namely a Caribbean Animal Resources Management (CARM) Program.
Radiopharmaceutical Treatment of Pheochromocytomas
Authors: SISSON JAMES C.
Published: July 2009

Abstract: Malignant pheochromocytomas, a group of tumors that include metastatic paragangliomas, often produce hypertension and episodic symptoms from secretion of norepinephrine and sometimes epinephrine. In addition, the tumors usually manifest progressive metastases. Blockade of alpha and beta adrenergic receptors will control blood pressure and symptoms, but reduction of the malignancy has been difficult to achieve. Meta‐iodobenzylguanidine (MIBG) follows the pathways of norepinephrine and, when labeled with 131‐I, will concentrate sufficiently in the pheochromocytoma to impart therapeutic radiation. More than 100 patients have received treatment with 131‐I‐labeled MIBG at multiple medical centers. Individual doses were 3.7 to 18.5 GBq (100 to >500 mCi), and many patients received several doses separated by a few months. Partial remissions, recorded as decreased tumor presence and tumor function, have been observed in one‐third or more of the treated patients. However, complete remissions are rare, and recurrence/progression within two years is the rule. Toxicity was generally modest and temporary. Subsequent chemotherapy increased the benefits attained by 131‐I MIBG, but, in a small series of patients, this combination did not further change the outcome. Nevertheless, selective radiation from 131‐I MIBG or a similar radiopharmaceutical could play a valuable role in treatments that combine several types of attacks on this recalcitrant malignancy.
A Twenty‐Year Trip through the Chromaffin Cell
Authors: GARCÍA ANTONIO G.
Published: July 2009

Abstract: Described here is the origin of a fruitful, long‐lasting, and friendly international group of scientists whose main interests are chromaffin cells and exocytosis. Meetings have been held every two years in different countries of Europe and North America, and in Japan, Australia, and Israel. We have no formal society or written rules, and our intention is good science and friendship. The first of our International Symposia on Chromaffin Cell Biology (ISCCB‐1) was held in Ibiza (Spain) in 1982, and the most recent (ISCCB‐11) was held in San Diego (USA) in 2001. These symposia are attended by 100‐150 scientists from most European countries, Japan, Israel, the United States, Canada, South America, and Australia. The interest in the chromaffin cell as a model for studying basic mechanisms of calcium signaling, cell‐cell communication, exocytosis, and membrane and vesicle trafficking has been wide; I predict that this interest will grow in the coming years.
Neuroendocrine Cell Type‐Specific and Inducible Expression of Chromogranin/Secretogranin Genes
Authors: MAHATA SUSHIL K. , MAHAPATRA NITISH R. , MAHATA MANJULA , O'CONNOR DANIEL T.
Published: July 2009

Abstract: The chromogranin/secretogranins (Cg/Sg) are a family of soluble, acidic proteins representing major constituents in secretory vesicle cores of virtually all neuroendocrine tissues. We and others have identified the cyclic adenosine monophosphate response element (CRE) as the crucial promoter element responsible for neuroendocrine cell type‐specific expression of the Cg/Sg genes. In addition to CRE, GC‐rich domains in chromogranin B (CgB) and serum response element (SRE) in secretogranin II (SgII) promoters appear to play important roles in neuroendocrine cell type‐specific expression of CgB and SgII genes. Nicotinic‐cholinergic and peptidergic chromaffin cell stimuli evoke catecholamine secretion and augment biosynthesis of Cg/Sg genes. These stimuli signal to CgA gene transcription through the CRE in cis and through protein kinase A, protein kinase C, and mitogen‐activated protein kinase and CRE‐binding protein in trans. In addition to CRE, a GC‐rich domain in CgB and SRE in SgII promoters also play important roles in mediating inducible expression of the CgB and SgII genes. We conclude that CRE, GC‐rich domains, and SRE are crucial determinants of both cell type‐specific and secretagogue‐inducible expression of the Cg/Sg genes.
Proteolytic Processing of Chromogranins Is Modified in Brains of Transgenic Mice
Authors: LASLOP ANDREA , BECKER ALEXANDRA , LINDBERG IRIS , FISCHER‐COLBRIE REINER
Published: July 2009

Abstract: In normal brain, secretogranin II and chromogranins A and S are extensively converted by endopeptidases to the peptides secretoneurin, GE‐19, and PE‐11, respectively. After genetic knockout of PC2 or its helper protein 7B2, but not after mutation of carboxypeptidase E, endoproteolytic processing decreased, as indicated by appearance of intermediate‐sized processing products.
Influence of the Anx7 (+/−) Knockout Mutation and Fasting Stress on the Genomics of the Mouse Adrenal Gland
Authors: SRIVASTAVA MEERA , KUMAR PREETI , LEIGHTON XIMENA , GLASMAN MIRTA , GOPING GERTRUDE , EIDELMAN OFER , POLLARD HARVEY B.
Published: July 2009

Abstract: The Anx7 gene codes for a Ca2+/GTPase with calcium channel and membrane fusion properties that has been proposed to regulate exocytotic secretion in chromaffin and other cell types. We have previously reported that the homozygous Anx7 (+/−) knockout mouse has an embryonically lethal phenotype. However, the viable heterozygous Anx7 (+/−) mouse displays a complex phenotype that includes adrenal gland hypertrophy, chromaffin cell hyperplasia, and defective IP3 receptor (IP3R) expression. To search for a molecular basis for this phenotype, we have used cDNA microarray technology and have challenged control and mutant mice with fed or fasting conditions. We report that in the absence of the Anx7/IP3R signaling system, the cells in the adrenal gland are unable to discriminate between the fed and fasted states, in vivo. In control chromaffin cells, fasting is accompanied by an increased expression of structural genes for chromaffin cell contents, including chromogranin A and B, and DβH. There are also genes whose expression is specifically reduced. However, the Anx7 (+/−) mutation results in sustained expression of these nutritionally sensitive genes. We hypothesize that the calcium signaling defect due to the missing IP3R may be responsible for the global effects of the mutation on nutritionally sensitive genes. We further hypothesize that the tonically elevated expression of chromogranin A, a reportedly master control “switch” for dense core granule formation, may contribute to the process driving glandular hypertrophy and chromaffin cell hyperplasia in the Anx7 (+/−) mutant mouse.
Phosphorylation of Tyrosine Hydroxylase in Isolated Mice Adrenal Glands
Authors: TOSKA KAREN , KLEPPE RUNE , COHEN PHILIP , HAAVIK JAN
Published: July 2009

Abstract: The site‐specific phosphorylation of tyrosine hydroxylase (TH) was studied in mouse adrenal tissue. On addition of arsenite, a rapid increase in Ser19 phosphorylation occurred, concomitant with phosphorylation of p38 protein kinase. This is consistent with studies in other mammals, indicating a role of stress‐activated protein kinases in TH regulation.
Cholinergic Activation of Glucose Transport in Bovine Chromaffin Cells Involves Calmodulin and Protein Kinase Cζ Signaling
Authors: SERCK‐HANSSEN GULDBORG , GRØNNING MONA , FLADEBY CATHRINE , SKAR ROBERT
Published: July 2009

Abstract: The aim of the present study was to delineate possible signaling pathways involved in acetylcholine (Ach)‐induced glucose transport in chromaffin cells, a widely applied model system for sympathetic neurons. Acute Ach stimulation (10 min) enhanced the rate of glucose transport through activation of both nicotinic and muscarinic receptors. The calmodulin antagonist, W13, and the protein kinase C (PKC) inhibitor, staurosporine, each partially depressed Ach‐induced glucose transport, with staurosporine exhibiting the stronger inhibitory effect. Pretreating the cells with phorbol 12‐myristate 13‐acetate (PMA) to downregulate PKC activity did not affect the nicotine‐induced glucose transport, but completely attenuated that activated by muscarine, suggesting that Ach activation of transport involved both diacylglycerol‐independent (PKCζ) and diacylglycerol‐dependent PKCs (PKCα/PKCε). The PI 3‐kinase inhibitor, wortmannin, diminished the Ach response, consistent with activation of the PKCs by the upstream PI 3‐kinase‐dependent phosphoinositide‐dependent kinase, PDK1. Cholinergic activation strongly activated the ERK1/ERK2 cascade and p38 MAP kinase, but only p38 MAP kinase appeared to play a role, however minor, in nicotine‐induced glucose uptake. The results are consistent with PKCs being more important than calmodulin in coupling cholinergic activation to glucose transport in chromaffin cells, but additional, yet unidentified, signaling pathways appear to be needed to obtain full activation of glucose transport in response to Ach.
Effects of Methyllycaconitine and Related Analogues on Bovine Adrenal α3β4* Nicotinic Acetylcholine Receptors
Authors: BRYANT DARRELL L. , FREE R. BENJAMIN , THOMASY SARA M. , LAPINSKY DAVID J. , ISMAIL K. A. , ARASON K. M. , BERGMEIER STEPHEN C. , McKAY DENNIS B.
Published: July 2009

Abstract: Adrenal secretion and binding studies were performed using ring E analogues of methyllycaconitine to assess structural determinants affecting activity on bovine adrenal α3β4* nicotinic receptors. The most potent analogues are as potent as many inhibitors of adrenal secretion. Our data support the potential use of methyllycaconitine analogues to generate nicotinic receptor subtype‐specific compounds.
Localization of Phosphatidylinositol 4,5‐P2 Important in Exocytosis and a Quantitative Analysis of Chromaffin Granule Motion Adjacent to the Plasma Membrane
Authors: HOLZ RONALD W. , AXELROD DANIEL
Published: July 2009

Abstract: A slow ATP‐dependent priming step precedes a rapid, Ca2+‐dependent triggering step in exocytosis in chromaffin cells and in most, if not all, differentiated secretory cells. A major component of ATP‐dependent secretion in permeabilized cells reflects the maintenance of the polyphosphoinositides, especially PtdIns‐4,5‐P2. Here we summarize recent experiments with PH‐GFP (binds to PtdIns‐4,5‐P2) that indicate that PtdIns‐4,5‐P2 is localized primarily on the plasma membrane in chromaffin cells, and that it is this pool that plays a role in exocytosis. It is demonstrated that transiently expressed PH‐GFP inhibits secretion in subsequently permeabilized cells. Recent studies using total internal reflection fluorescent microscopy (TIRFM) to measure chromaffin granule motion adjacent to the plasma membrane are also summarized. The quantitative analysis indicates that chromaffin granule motion is highly restricted and suggests that chromaffin granules are caged or tethered immediately adjacent to the plasma membrane.
Rim and Exocytosis: Rab3a‐Binding and Secretion‐Enhancing Domains Are Separate and Function Independently
Authors: SUN LEI , BITTNER MARY A. , HOLZ RONALD W.
Published: July 2009

Abstract: Rim1 has been identified in brain by its ability to bind Rab3a‐GTP and has been postulated to be a Rab3a effector protein. Like Rabphilin3, another putative Rab3a effector protein, Rim1 modulates secretion and contains a zinc‐finger and two C2 domains. We have investigated the structural basis for the ability of Rim1 to bind Rab3a‐GTP and to stimulate exocytosis in chromaffin cells.
Exocytosis Studies in a Chromaffin Cell‐Free System
Authors: WIEGAND ULRICH K. , DON‐WAUCHOPE ANDREW , MATSKEVICH IOULIA , DUNCAN RORY R. , GREAVES JENNIFER , SHIPSTON MICHAEL J. , APPS DAVID K. , CHOW ROBERT H.
Published: July 2009

Abstract: We have developed a system for the real‐time study of regulated exocytosis in living, cultured bovine adrenal chromaffin cells (BCCs). Exocytosis was monitored by the use of total internal reflection fluorescence (TIRF) microscopy to image single large dense‐core secretory vesicles (LDCVs). Fluorescent labeling of LDCVs was achieved either with the membrane‐permeant weak base, acridine orange (AO), or by transduction of BCCs so as to express a fluorescent chimeric “cargo” protein that is targeted to LDCVs. In either case, exocytosis is visible by the disappearance of a vesicle accompanied by a bright flash as the fluorescent contents leave the acidic LDCV lumen, move towards the source of the evanescent wave, and disperse into the extracellular medium. Furthermore, for the first time, we have developed a broken‐cell system for real‐time imaging in BCCs, in which individual plated cells are mechanically “unroofed” with a jet of intracellular medium, leaving a membrane patch with docked vesicles on the coverslip. In this cell‐free system, a subpopulation of docked granules undergoes exocytosis in response to calcium. This approach allows us direct experimental access to membrane‐docked LDCVs in order to investigate the dependence of exocytosis on defined protein components and intracellular conditions at the single‐vesicle level. In addition, this system can be used for a reconstitution analysis of the exocytosis machinery. Finally, we demonstrate the use of 2D+1 image analysis for visualizing single‐vesicle exocytosis. We use this approach for a rapid analysis of larger numbers of imaged vesicles.
Neuropeptides and Chromogranins
Authors: LASLOP ANDREA , MAHATA SUSHIL K.
Published: July 2009

Abstract: In large dense core vesicles, also referred to as chromaffin granules in adrenomedullary chromaffin cells, transmitters or hormones are stored together with neuropeptides and chromogranins. For most neuropeptides, functions have been established and new findings on their secretion, receptors, and synthesis regulation are reported. The functions of chromogranins are less clear, and possible roles as peptide precursors, Ca2+ regulators, inducers of secretory granule biogenesis, and as nuclear constituents are discussed.
WE‐14, a Chromogranin A‐Derived Neuropeptide
Authors: CURRY W. J. , BARKATULLAH S. C. , JOHANSSON A. N. , QUINN J. G. , NORLEN P. , CONNOLLY C. K. , McCOLLUM A. P. , McVICAR C. M.
Published: July 2009

Abstract: The neuropeptide WE‐14 is derived from the posttranslational processing of chromogranin A (CgA). While CgA is expressed in a preponderance of neuroendocrine cells, WE‐14 is generated in a distinct subpopulation of CgA‐immunopositive cells, most notably in the adrenal, pituitary, and parathyroid glands. Physiological and pharmacological studies have demonstrated that CgA is cleaved to generate WE‐14 in the adrenal chromaffin cell population and in the enterochromaffin‐like (ECL) cells of the oxyntic mucosa. Pathological analyses of neuroendocrine tumors have revealed a heterogeneous pattern of WE‐14 immunostaining, with variable concentrations quantified and chromatographically resolved in tissue extracts. Phylogenetic surveys have demonstrated that WE‐14 exhibits an ancient lineage, while ontogenetic examination has shown that it is generated at an early stage during fetal development. Putative WE‐14 receptor binding sites have been identified in several tissues; however, the physiological role of WE‐14 remains enigmatic.
Genetic Basis of Clinical Catecholamine Disorders
Authors: GARLAND EMILY M. , HAHN MAUREEN K. , KETCH TERRY P. , KELLER NANCY R. , KIM CHUN‐HYUNG , KIM KWANG‐SOO , BIAGGIONI ITALO , SHANNON JOHN R. , BLAKELY RANDY D. , ROBERTSON DAVID
Published: July 2009

Abstract: Norepinephrine and epinephrine are critical determinants of minute‐to‐minute regulation of blood pressure. Here we review the characterization of two syndromes associated with a genetic abnormality in the noradrenergic pathway. In 1986, we reported a congenital syndrome of undetectable tissue and circulating levels of norepinephrine and epinephrine, elevated levels of dopamine, and absence of dopamine‐β‐hydroxylase (DBH). These patients appeared with ptosis and severe orthostatic hypotension and lacked sympathetic noradrenergic function. In two persons with DBH deficiency, we identified seven novel polymorphisms. Both patients are compound heterozygotes for a variant that affects expression of DBH protein via impairment of splicing. Patient 1 also has a missense mutation in DBH exon 2, and patient 2 carries missense mutations in exons 1 and 6. Orthostatic intolerance is a common syndrome affecting young women, presenting with orthostatic tachycardia and symptoms of cerebral hypoperfusion on standing. We tested the hypothesis that abnormal norepinephrine transporter (NET) function might contribute to its etiology. In our proband, we found an elevated plasma norepinephrine with standing that was disproportionate to the increase in levels of dihydroxphenylglycol, as well as impaired norepinephrine clearance and tyramine resistance. Studies of NET gene structure revealed a coding mutation converting a conserved alanine residue in transmembrane domain 9 to proline. Analysis of the protein produced by the mutant cDNA demonstrated greater than 98% reduction in activity relative to normal. The finding of genetic mutations responsible for DBH deficiency and orthostatic intolerance leads us to believe that genetic causes of other autonomic disorders will be found, enabling us to design more effective therapeutic interventions.
Analysis of Signaling Pathways Using Functional Proteomics
Authors: RESING KATHERYN A.
Published: July 2009

Abstract: Advances in analytical methods for protein analysis by mass spectrometry provide new tools for global analysis of the expressed protein profile of cells (referred to as proteomics). Currently, available methodology samples only part of the proteome. This is sufficient for analysis of signal transduction, because signaling pathways contain enzymes, which modify high‐abundance proteins other than those of the pathway. Thus, modulation of the signaling through a pathway will produce a “footprint” in the proteome that is characteristic of a specific cell phenotype. Comparison of different samples to identify these differences in posttranslational modification or protein expression is referred to as functional proteomics. This review surveys the methods in widest use in functional proteomics, as well as a few promising new ones. Although proteomic analyses were first conducted 26 years ago, a renewed interest is fueled by several recent advances. Most important are the availability of public genome and protein databases and the development of high‐sensitivity, easy‐to‐use mass spectrometers and database search engines capable of exploiting these databases. Other important advances include improved two‐dimensional polyacrylamide gel electrophoresis (2D‐PAGE), computer programs for analysis of the 2D‐PAGE gel images, protocols for proteolytic digestion of proteins in excised gel pieces, and low‐flow chromatography methods. Despite the limitations of these methods, they can distinguish subtle changes in the phenotype of cells, providing the basis for future studies in regulation of the phenotype.
The Association of Vesicular Contents and Its Effects on Release
Authors: WIGHTMAN R. MARK , TROYER KEVIN P. , MUNDORF MICHELLE L. , CATAHAN REBECCA
Published: July 2009

Abstract: The oxidation of catecholamines with a carbon‐fiber electrode can be used to monitor exocytosis at the single cell level at a variety of different types of cells. These measurements allow release to be followed from individual vesicles and have revealed several unique aspects concerning the coupling between release and storage. The strong association of the vesicular components in chromaffin cells dictates the time course of extrusion of the vesicle contents. Furthermore, liberation of the Ca2+ normally stored within the vesicles can promote exocytosis without an external Ca2+ source.