bluuebunny/crossref_metadata_2025 huggingface dataset:
Nitric Oxide
Authors: MOHANAKUMAR KOCHUPURACKAL P. , THOMAS BOBBY , SHARMA SUDARSHANA M. , MURALIKRISHNAN DHANASEKHARAN , CHOWDHURY RUKHSANA , CHIUEH CHUANG C.
Published: July 2009

Abstract: Indirect evidence, including neuroprotection against 1‐methyl‐4‐phenyl‐1,2,3,6‐tetrahydropyridine (MPTP)‐neurotoxicity by nitric oxide synthase (NOS) inhibitors and resistance of transgenic animals deficient in NOS, is controversial. We have reviewed evidence in favor of oxidative stress during the development of MPTP‐neurotoxicity and the influence of antioxidants, including nitric oxide (NO) and NO donors, on MPTP‐induced dopaminergic neurotoxicity. Systemic administration of MPTP causes dose‐dependent generation of hydroxyl radicals (OH) in vivo in the striatum in mice; OH scavengers protect dopaminergic neurons from this insult. On the other hand the role of NO in MPTP‐neurotoxicity is controversial. Hitherto, no direct evidence for the involvement of NO in MPTP neurotoxicity has been available. MPTP does not affect inducible‐NOS mRNA level or its expression in SN or the striatum. Nitroglycerine, a NO donor, can attenuate MPTP‐induced dopamine depletion in the striatum by virtue of its OH scavenging action. Several other NO donors have also been shown to scavenge the OH generated, following Fenton chemistry in vitro, and to protect against in vivo dopaminergic neurotoxicity by small mass iron complex formation. This evidence suggests that NO renders protection against MPTP‐induced OH‐mediated nigrostriatal lesions, acting as an antioxidant.
Nitric Oxide Triggers Classic Ischemic Preconditioning
Authors: LOCHNER AMANDA , MARAIS ERNA , TOIT EUGENE DU , MOOLMAN JOHAN
Published: July 2009

Abstract: The role of NO in the classic ischemic preconditioning phenomenon of the myocardium is not well defined, and was investigated by using the isolated perfused rat heart as a model. Hearts were preconditioned with 3 × 5 minute ischemia in the presence and absence of the NOS inhibitors l‐NAME (50 μM) and l‐NNA (50 μM), and the guanylyl cyclase inhibitor ODQ (20 μM). These inhibitors significantly attenuated the protective effect of preconditioning against 25‐min global ischemia (as measured by functional recovery), specifically if administered during the triggering phase. Cyclic infusions (3 × 5 min) of the NO‐donors SNAP (50 μM) and SNP (100 μM) elicited protection against both 25‐min global or low‐flow ischemia. Hearts preconditioned with NO donors displayed significantly superior functional reserve, if stimulated with adrenaline, compared to hearts preconditioned with ischemia. Although the NO donors SNAP and SNP both activated p38 MAPK during the preconditioning protocol, protection was accompanied by significantly decreased p38 MAPK activity during sustained ischemia, as was the case in ischemic preconditioning. We conclude that (1) NO is a trigger for classic preconditioning, (2) cGMP generation plays an important role in its protection, (3) attenuation of p38 MAPK during sustained ischemia accompanies NO preconditioning and may mediate cardiac protection, and (4) preconditioning with NO may be more advantageous than using ischemia.
Neural‐Immune Interactions in Health and Disease
Authors: ESKANDARI FARIDEH , STERNBERG ESTHER M.
Published: July 2009

Abstract:Many lines of research have established the numerous routes by which the immune and central nervous systems (CNS) communicate. The CNS signals the immune system via hormonal and neuronal pathways and the immune system signals the CNS through similar routes via immune mediators and cytokines. The primary hormonal pathway by which the CNS regulates the immune system is the hypothalamic‐pituitary‐adrenal (HPA) axis, through the hormones of the neuroendocrine stress response. The sympathetic nervous system regulates immune system function primarily via adrenergic neurotransmitters released through neuronal routes. Neuroendocrine regulation of immune function is essential for survival during stress or infection and to modulate immune responses in inflammatory disease. Glucocorticoids are the main effector endpoint of the neuroendocrine response system.
Immune Homeostasis Requires Several Biologic Factors Including Glucocorticoid Hormones
Authors: CANCEDDA CORRADO , FILACI GILBERTO , PUPPO FRANCESCO , GHIO MASSIMO , CONTINI PAOLA , INDIVERI FRANCESCO
Published: July 2009

Abstract: Immunological tolerance can be achieved by several mechanisms including suppressor cells, soluble factors, and neurohormonal mediators. On the cellular level, we isolated a population of CD8+CD28− T cells capable of inhibiting anti‐CD3 mAb‐induced proliferation of autologous peripheral blood mononuclear cells in an HLA‐I nonrestricted manner via production of IFN‐γ and IL‐6. Interestingly, CD8+CD28− T cells from systemic lupus erythematosus patients with active disease do not display this inhibitory activity and show a marked imbalance between inhibitory (IL‐6) and stimulatory (IL‐12) cytokines. For soluble factors, we studied soluble HLA molecules (sHLAs) and double‐stranded DNA (ds‐DNA). Soluble HLA‐I (sHLA‐I) molecules induce soluble Fas ligand (sFasL) secretion and trigger apoptosis in phytohemagglutin (PHA)‐activated Fas+ T cells. Double‐stranded DNA binds to HLA‐II molecules and inhibits HLA‐II‐mediated antigen presentation. On the neurohormonal side, we focused our attention on the immunological activity of corticosteroids (CTSs). CTSs inhibit recirculation of CD4+ T cells, suppress the proliferation and immunological function of activated T cells, and induce apoptosis of activated lymphocytes. Taken together, these data suggest the presence of a complex network of immunoregulatory mechanisms in which CTSs play a strong role supporting their recognized efficacy in the treatment of inflammatory and immunological diseases.
Differential Male and Female Adrenal Cortical Steroid Hormone and Cortisol Responses to Interleukin‐6 in Humans
Authors: SILVA CÂNDIDA , INÊS LUIS SOUSA , NOUR DOLORES , STRAUB RAINER H. , DA SILVA JOSÉ ANTÓNIO P.
Published: July 2009

Abstract: Evidence from experimental animal studies show that sex hormones influence the glucocorticoid response to a variety of inflammatory and noninflammatory stimuli. In this study we assessed gender differences in the response of ACTH and cortisol in normal young male and female humans following intravenous infusion of human IL‐6 in various dosages. Males presented a significantly stronger ACTH production in response to IL‐6 than females. Peak cortisol response, however, was similar in males and females. Cortisol/ACTH ratios were significantly higher in females than in males, both at baseline and after each of the IL‐6 dosages. These results suggest that an effective glucocorticoid response requires similar levels of IL‐6 in males and females. However, they also suggest that the adrenals of males and females have different sensitivities to ACTH (higher in females) and possibly also to direct IL‐6 stimulation.
Interactions between Glucocorticoids and Cytokines in the Bone Microenvironment
Authors: ANGELI A. , DOVIO A. , SARTORI M. L. , MASERA R. G. , CEOLONI B. , PROLO P. , RACCA S. , CHIAPPELLI F.
Published: July 2009

Abstract: Cytokines belonging to the so‐called interleukin‐6 (IL‐6) or gp130 cytokine family, notably IL‐6 and IL‐11, are known as pro‐resorptive cytokines, in that they promote osteoclastogenesis. Glucocorticoid (GC)‐induced osteoporosis is admittedly the most frequent secondary osteoporosis. The pathogenesis still has many unresolved issues. Although the effects of GCs on cytokine production and recognition have been extensively studied, little is known about the effects of cytokines on GC action at the target level. We have focused on the effects of IL‐6 and IL‐11 on specific binding by type II GC receptors (GRs) in two human osteoblast‐like cell lines (Saos‐2 and MG‐63) that have remarkably different constitutive expression of these cytokines and GRs as well. We have provided evidence that IL‐6 upregulates GR binding sites, while IL‐11 downregulates these sites, as determined by radioligand binding assay and Scatchard analysis. GR affinity (Kd) did not change after exposure to both cytokines. A number of experiments were consistent with the view that in human osteoblast‐like cells, cytokines of the IL‐6 family have autocrine modulatory effects on GRα (GRβ is a variant that does not bind specifically in our method). Complex effects of GCs on the system(s) of proinflammatory/anti‐inflammatory cytokines and conversely of these cytokines on GC action could account for the dynamics of bone loss in patients given GCs and conceivably having high concentrations of these cytokines in the bone microenvironment.
Estrogens and the Vascular Endothelium
Authors: CID MARIA C. , SCHNAPER H. WILLIAM , KLEINMAN HYNDA K.
Published: July 2009

Abstract: Estrogens exert important regulatory functions on vessel wall components, which may contribute to the increased prevalence and severity of certain chronic inflammatory and autoimmune diseases in females and the lower cardiovascular risk observed in premenopausal women. Endothelial cells have been recently identified as targets for estrogens, and estrogen receptors have been demonstrated in endothelial cells from various vascular beds. This review focuses on the regulatory function of estrogens in endothelial cell responses relevant to vessel inflammation, injury, and repair; estrogen effects on nitric oxide production and release; estrogen modulation of endothelial cell adhesion molecule expression; and estrogen regulation of angiogenesis. The mechanisms through which estrogen regulates endothelial cell functions are complex and involve both genomic and nongenomic mechanisms.
Influence of Cytokines and Growth Factors on Distinct Steroidogenic Enzymesin Vitro
Authors: HERRMANN M. , SCHÖLMERICH J. , STRAUB R. H.
Published: July 2009

Abstract:Cytokines (IL‐1, IL‐6, IL‐8, IL‐11, TNF, IFN‐γ, and TGF‐β) and growth factors (EGF, bFGF, aFGF, and KGF) play an important role in modulation of hormone secretion by directly influencing specific enzyme steps of steroidogenesis in various endocrine cell types. For this tabular data collection, the following enzyme steps were considered: steroidogenic acute regulatory protein (StAR), side chain cleavage enzyme (P450scc), 3 beta‐hydroxysteroid dehydrogenase, 17‐alpha‐hydroxylase/17,20‐lyase (P450c17), 17‐beta‐hydroxysteroid‐dehydrogenase, aromatase complex, 5‐alpha‐reductase, P450c21, DHEAS sulfatase, and DHEA sulfotransferase. This collection summarizes the current information on how the mentioned cytokines and growth factors influence particular enzyme steps.
Hormone Replacement Therapy and Chronic Graft‐versus‐Host Disease Activity in Women Treated with Bone Marrow Transplantation for Hematologic Malignancies
Authors: BALLEARI ENRICO , GARRÈ SIMONA , VAN LINT MARIA TERESA , SPINELLI SIMONETTA , CHIODI SANDRA , REPETTO ENRICO , MASSA GIULIO , BACIGALUPO ANDREA , GHIO RICCARDO
Published: July 2009

Abstract: Several lines of experimental evidence suggest that sex hormones may influence the development and activity of chronic graft‐versus‐host disease (cGVHD), which frequently occurs in patients undergoing allogeneic bone marrow transplantation (ABMT). Following ABMT, young women are commonly treated with hormone replacement therapy (HRT) because of irreversible gonadal failure. It seemed therefore worthwhile to investigate the effects of this therapy on the activity of cGVHD. Premenopausal women treated with ABMT for hematological malignancies between January 1997 and December 2000 were evaluated for cGVHD activity. They were divided into two groups, depending on whether or not they were treated with HRT. Seventy‐one women qualified for the present study: 39 received HRT (treated group), while 32 did not (controls). In both groups of patients, cGVHD activity score was comparable before the start of HRT. No differences were observed in cGVHD activity score between the HRT group and controls after 3, 6, 12, and 24 months from the start of HRT. Furthermore, HRT did not induce any increase in the cGVHD activity score in the treated group of patients at any time from the start of HRT. According to present data, HRT did not appear to influence the activity of cGVHD in young women who underwent ABMT for hematological malignancies. Therefore, we can safely propose this therapy for women with gonadal failure after ABMT.
Delayed Neutrophil Apoptosis Induced by Synovial Fluid in Rheumatoid Arthritis
Authors: OTTONELLO LUCIANO , FRUMENTO GUIDO , ARDUINO NICOLETTA , BERTOLOTTO MARIA , MANCINI MARINA , SOTTOFATTORI ENZO , DALLEGRI FRANCO , CUTOLO MAURIZIO
Published: July 2009

Abstract: The fate of neutrophils at sites of inflammation, where these cells are likely exposed to both anti‐ and proapoptotic influences, needs to be clarified. To investigate this issue, we studied the survival of neutrophils in the presence of articular fluids from RA joints before and after immune complex activation. Eight of eleven samples of RA synovial fluid studied were found to inhibit spontaneous and immune complex‐stimulated neutrophil apoptosis. No relationships were found between GM‐CSF and TNF‐α concentrations measured on each sample of synovial fluid studied and the levels of neutrophil apoptosis detectable in the presence of the same synovial fluid. Furthermore, no activity on neutrophil survival was observed at either physiologic or pharmacologic concentrations of estradiol. On the contrary, the synovial fluid anti‐apoptotic activity correlates (r2= 0.8818, p < 0.0001) with the adenosine detected at concentrations in each sample ranging from 18.7 to 52.4 μM. Finally, synovial fluids were incapable of interfering with neutrophil activation evaluated as superoxide anion production. Our results suggest that the microenvironment of rheumatoid synovial fluid is a proinflammatory milieu responsible for the in loco persistence of activated and long‐surviving neutrophils.
Antiproliferative‐Antiinflammatory Effects of Methotrexate and Sex Hormones on Cultured Differentiating Myeloid Monocytic Cells (THP‐1)
Authors: CUTOLO MAURIZIO , SULLI ALBERTO , CRAVIOTTO CHIARA , FELLI LAMBERTO , PIZZORNI CARMEN , SERIOLO BRUNO , VILLAGGIO BARBARA
Published: July 2009

Abstract: Methotrexate (MTX) is believed to exert both antiproliferative and antiinflammatory effects in a dose‐related manner in a majority of rheumatoid arthritis (RA) patients along with an abrupt flare of the disease after drug discontinuation. To investigate the antiproliferative and antiinflammatory actions of MTX and the combined action of sex hormones, we evaluated these effects in differentiated monocytic myeloid cells (THP‐1) prestimulated with testosterone (T) or 17‐β estradiol (E2). The effects of MTX and T combined treatment (T/MTX) on THP‐1 cells showed a significant inhibition of cell proliferation when compared with E2/MTX‐ treated cells or controls: 53% at 72 h versus E2‐treated cells; 58% at 96 h versus E2‐treated cells; and 41% versus controls, respectively. Bax and Fas CD95 expression was found increased in T‐treated cells: 14% T at 48 h vs. E2‐treated cells and controls; 45% T at 72 h versus E2‐treated cells and controls; 97% at 96 h versus E2‐treated cells and 37% versus controls for Bax: 33%, 41%, and 42% T versus E2‐treated cells for Fas. Moreover, a significant decrease of IL‐12 levels in T/MTX treated cells was found at any time when compared to E2‐treated cells. In summary, the association of testosterone and MTX compared to MTX alone suggests possible synergistic actions. Therefore, the enhancing antiinflammatory effects exerted by androgens might represent a further explanation for the reduced frequency of inflammatory diseases in male subjects.
Serum Prolactin Concentrations in Male Patients with Rheumatoid Arthritis
Authors: SERIOLO BRUNO , FERRETTI VALENTINO , SULLI ALBERTO , FASCIOLO DANIELA , CUTOLO MAURIZIO
Published: July 2009

Abstract: Altered serum prolactin (PRL) levels have been reported in autoimmune diseases; however, data of serum PRL concentrations in rheumatoid arthritis (RA) are contradictory. We evaluated the PRL status in men affected by RA and the possible relationships among serum PRL levels, bone mass, and disease activity. We investigated 29 men affected by RA and 30 age‐ and sex‐matched controls. All patients were evaluated for serum PRL levels, parameters of disease activity, and bone mineral density (BMD) at L2‐L4 and the femoral neck. Serum PRL levels were found significantly higher in men with RA than in controls (p= 0.001). High serum PRL levels were significantly correlated with duration of RA and some laboratory parameters of RA disease activity. A negative correlation between femoral BMD and serum PRL levels were found (r=−0.821, p= 0.001). Male patients affected by RA showed high serum PRL levels. The serum PRL concentration was found to be increased in relation to the duration and the activity of the disease. Serum PRL levels do not seem to have any relationship with the BMD, at least in RA.
Melatonin Serum Levels in Rheumatoid Arthritis
Authors: SULLI A. , MAESTRONI G.J. M. , VILLAGGIO B. , HERTENS E. , CRAVIOTTO C. , PIZZORNI C. , BRIATA M. , SERIOLO B. , CUTOLO M.
Published: July 2009

Abstract:The pineal hormone melatonin (MLT) exerts a variety of effects on the immune system. MLT activates immune cells and enhances inflammatory cytokine and nitric oxide production. Cytokines are strongly involved in the synovial immune and inflammatory response in rheumatoid arthritis (RA) and reach the peak of concentration in the early morning, when MLT serum level is higher. Nocturnal MLT serum levels were evaluated in 10 RA patients and in 6 healthy controls. Blood samples were obtained at 8 and 12 p.m., as well as at 2, 4, 6, and 8 a.m. MLT serum levels at 8 p.m. and 8 a.m. were found to be higher in RA patients than in controls (p< 0.05). In both RA patients and healthy subjects, MLT progressively increased from 8 p.m. to the first hours of the morning, when the peak level was reached (p< 0.02). However, MLT serum level reached the peak at least two hours before in RA patients than in controls (p< 0.05). Subsequently, in RA patients, MLT concentration showed a plateau level lasting two to three hours, an effect not observed in healthy controls. After 2 a.m., MLT levels decreased similarly in both RA patients and healthy subjects. Several clinical symptoms of RA, such as morning gelling, stiffness, and swelling, which are more evident in the early morning, might be related to the neuroimmunomodulatory effects exerted by MLT on synovitis and might be explained by the imbalance between cortisol serum levels (lower in RA patients) and MLT serum levels (higher in RA patients).
Role of Interleukin‐6 in Stress Response in Normal and Tumorous Adrenal Cells and during Chronic Inflammation
Authors: WILLENBERG HOLGER S. , PÄTH GÜNTER , VÖGELI THOMAS A. , SCHERBAUM WERNER A. , BORNSTEIN STEFAN R.
Published: July 2009

Abstract:Interleukin‐6 (IL‐6) is the end‐product of a cytokine signaling cascade and is secreted by specialized immune cells during inflammation. It has a great influence on many functions, including differentiation, stimulation, and activation of immune cells, or other cells of neuroendocrine origin. Thus, IL‐6 serves as a key messenger in its communication with the neuroendocrine system, and serves as a potent activator of the hypothalamic‐pituitary‐adrenal axis at all levels. Changes in the levels of expression of this cytokine and its receptor have been observed during chronic inflammatory disease, and have been associated with tumorigenesis. Therefore, we studied the effect of IL‐6 on normal and adenomatous human adrenal cellsin vitro. The expression of IL‐6 receptor mRNA was quantified within the same tissue. IL‐6 potently stimulated cortisol secretion from dispersed normal human adrenal cells. We found immunoreactivity for the IL‐6 receptor on cultured cells and paraffin‐embedded sections of adrenal tissues. Further, there was a more pronounced expression of IL‐6 mRNA in adrenal adenomas of patients with Cushing's syndrome, compared to normal human adrenals. Despite this fact, the sensitivity of cells of adenomatous adrenal glands to IL‐6 was significantly decreased relative to cells from normal controls. These results were confirmed employing the permanent adrenocortical cancer cell line model NCI‐H295. We infer that the loss of responsivity of tumorous adrenal cells to IL‐6, and in part corticotropin, is an important step in the process of adrenal tumorigenesis by which regulation by differentiating proteins is bypassed.
Effects of Chronic Food Restriction Stress and Chronic Psychological Stress on the Development of Adjuvant Arthritis in Male Long Evans Rats
Authors: ŠEREŠ J. , STANČÍKOVÁ M. , ŠVÍK K. , KRSOVÁ D. , JURČOVIČOVÁ J.
Published: July 2009

Abstract: The aim of the present experiments was to study the effect of stress of chronic food restriction (FR) and of repeated psychological challenge (PS) on the development of adjuvant arthritis in the Long Evans male rats. In the FR series, four groups of animals were compared: non‐treated control (C) and arthritic (AA) rats, both with free access to food and water, and two analogous groups with a 40% food restriction—FR and AA‐FR. All animals were killed 22 days after injection of cFA. In the PS series, stress was induced by random daily exposures of the rats to isolation, over‐crowding, food/water deprivation, foot shock, tilting, fear for 14 days before cFA injection and 12 days thereafter (groups: C, AA, PS, and AA‐PS). Arthritis causes swelling of the hindpaw, which was prevented in the AA‐FR group. PS causes more severe disease symptoms: AA‐PS rats had more severe hindpaw swelling than AA rats. Forty percent food restriction associated with elevated CORT levels mitigated inflammatory parameters activated during AA. PS worsened the disease. These results suggested that activated CORT is not the only cause of disease suppression, but some metabolic changes during FR play a role.
Mechanisms of Pain in Arthritis
Authors: SCHAIBLE HANS‐GEORG , EBERSBERGER ANDREA , VON BANCHET GISELA SEGOND
Published: July 2009

Abstract: Inflammation in the joint causes peripheral sensitization (increase of sensitivity of nociceptive primary afferent neurons) and central sensitization (hyperexcitability of nociceptive neurons in the central nervous system). The processes of sensitization are thought to be the basis of arthritic pain that appears as spontaneous pain (joints at rest) and hyperalgesia (augmented pain response on noxious stimulation and pain on normally nonpainful stimulation). Sensitization also facilitates efferent neuronal processes through which the nervous system influences the inflammatory process. Peripheral sensitization is produced by the action of inflammatory mediators such as bradykinin, prostaglandins, neuropeptides, and cytokines which activate corresponding receptors in proportions of nerve fibers. In addition, the expression of receptors, for example, bradykinin and neurokinin 1 receptors, is upregulated during inflammation. The development of hyperexcitability of spinal cord neurons is produced by various transmitter/receptor systems that constitute and modulate synaptic activation of the neurons. The key transmitter is glutamate that activates N‐methyl‐d‐aspartate (NMDA) and non‐NMDA receptors on spinal cord neurons. Blockade of these receptors prevents and reduces central sensitization. Excitatory neuropeptides (substance P and calcitonin gene‐related peptide) further central sensitization. Central sensitization also is facilitated by mediators that have complex actions (e.g., prostaglandin E2). Spinal PGE2 binds to receptors at presynaptic endings of primary afferent neurons (thus influencing synaptic release) and to receptors on postsynaptic spinal cord neurons. The administration of PGE2 to the spinal cord surface produces changes of responsiveness of spinal neurons similar to peripheral inflammation, and spinal indomethacin to the spinal cord attenuates development of hyperexcitability significantly.
Expression of IL‐15 and IL‐15 Receptor Isoforms in Select Structures of Human Fetal Brain
Authors: KUROWSKA M. , RUDNICKA W. , MAŚLIŃSKA D. , MAŚLIŃSKI W.
Published: July 2009

Abstract: IL‐15, a key cytokine linking innate and acquired immunity, is expressed in many cell types and tissues. Recent data indicate constitutive expression of IL‐15 in human neural cell lines and tissues. The aim of the present study was to examine the expression patterns of mRNA encoding IL‐15 and IL‐15 receptor α (IL‐15Rα) isoforms in select structures of human fetal brain. We report that mRNA for IL‐15 and IL‐15Rα isoforms were expressed in all tested brain structures: cerebral cortex, cerebellum, hippocampus, and thalamus. However, the levels of IL‐15 and IL‐15Rα mRNA were higher in the hippocampus and cerebellum in comparison with cortex and thalamus. Moreover, higher levels of cytosol in comparison with membrane‐bound IL‐15 isoform were present in all brain structures. The constitutive, but distinct, expression of IL‐15 and its receptors in select human fetal brain structures suggests that IL‐15 plays a role in their development and physiology.
Carotid Atherosclerosis in Patients with Rheumatoid Arthritis
Authors: RODRIGUEZ G. , SULLI A. , CUTOLO M. , VITALI P. , NOBILI F.
Published: July 2009

Abstract: A preliminary study was undertaken to investigate the severity of atherosclerotic lesions of neck arteries in patients with rheumatoid arthritis (RA) in comparison with matched controls. The working hypothesis derives from the recent view that the atherosclerotic plaque is essentially an inflammatory lesion. Thus, patients affected by a chronic inflammatory disease, such as RA, might show acceleration of atherosclerosis despite treatment with anti‐inflammatory drugs. In 19 patients with RA the prevalence of atherosclerosis of internal carotid arteries, as seen on high‐resolution duplex‐scanning of neck arteries, was significantly (p < 0.05) higher than in 19 controls, whereas no significant difference was found for the other arteries. Factors underlying this finding are not known; increased levels of homocysteine and other thiol compounds that may enhance atherosclerosis in RA patients deserve further investigation. Moreover, more specific factors of RA are currently being investigated. In fact, immune complexes fixing C1q bind to endothelial receptors, thus triggering an upregulation of adhesion molecules, such as E‐selectin and intercellular and vascular cell adhesion molecules 1, on the endothelium surface.
Prolactin/Cortisol Ratio in Rheumatoid Arthritis
Authors: ZOLI ANGELO , FERLISI ESTER MARIA , LIZZIO MARCO , ALTOMONTE LORENZO , MIRONE LUISA , BARINI ANGELA , SCUDERI FLAVIA , BARTOLOZZI F. , MAGARO MARIO
Published: July 2009

Abstract: Prolactin (PRL) and glucocorticoids are hormones involved in the regulation of the immune system. Rheumatoid arthritis (RA) is an inflammatory condition that presents a diurnal rhythm of disease activity. PRL/cortisol ratio, and IL‐1β and TNF‐α levels were determined in patients with RA and in control subjects at 0600, 1000, 1400, 1800, 2200, and 0200 hours. In patients with RA we observed higher PRL/cortisol ratio at 0200 hours, whereas IL‐1β and TNF‐α reached their highest serum levels at 0200 and 0600 hours. In patients with RA we observed an imbalance in favor of proinflammatory hormones as opposed to levels of antiinflammatory hormones during nocturnal hours together with increased levels of IL‐1β and TNF‐α of the diurnal rhythm of disease activity.
Mutations of the Gene Encoding the Protein Kinase A Type I‐α Regulatory Subunit (PRKAR1A) in Patients with the “Complex of Spotty Skin Pigmentation, Myxomas, Endocrine Overactivity, and Schwannomas” (Carney Complex)
Authors: STRATAKIS CONSTANTINE A.
Published: July 2009

Abstract: Carney complex (CNC) is a familial multiple neoplasia syndrome associated with abnormal skin and mucosal pigmentation. The complex has features overlapping those of McCune‐Albright syndrome (MAS) and the other multiple endocrine neoplasias (MENs). CNC is inherited as an autosomal dominant trait, and the responsible genes have been mapped by linkage analysis to loci at 2p16 and 17q22‐24. Because of its unusual biochemical features (e.g., paradoxical responses to various endocrine signals) and its clinical similarities to MAS, genes implicated in cyclic nucleotide‐dependent signaling, including GNAS1 (which is responsible for MAS), had been considered likely candidates for causing CNC. The gene encoding the protein kinase A (PKA) type I‐α regulatory subunit (RIα), PRKAR1A, had been mapped to 17q22‐24; loss‐of‐heterozygosity (LOH) analysis using polymorphic markers from this region revealed consistent changes in tumors from patients with CNC, including those from one family previously mapped to 17q22‐24. Investigation of a polymorphic site within the 5′ of the PRKAR1A gene showed segregation with the disease and retention of the allele bearing the disease gene in CNC tumors. Mutations of the PRKAR1A gene were also found to have occurred de novo in sporadic cases of CNC; no mutations were found in kindreds mapping to 2p16. Thus, genetic heterogeneity in CNC was confirmed; in total, 41% of all patients with CNC had mutations in the PRKAR1A gene. All mutations were frameshifts, insertions, and deletions that led to nonsense mRNA and premature termination of the predicted peptide product. Functional studies in CNC tumors suggested that inactivating mutations of the PRKAR1A gene led to nonsense mRNA decay (the mutant peptide product was not present) and were associated with dysregulated PKA activity, increased responsiveness to cAMP, and excess of type‐II PKA activity. We conclude that the PRKAR1A gene, coding for the RIα subunit of PKA, a critical cellular component of a number of cyclic nucleotide‐dependent signaling pathways, is mutated in a subset of patients with CNC. In their tumors, there is LOH of the normal allele, suggesting that normal RI‐α may have tumor suppression function in the tissues affected by CNC. An excess of type‐II PKA activity was present in affected tissues, which may be responsible for the apparent tumorigenicity of PRKAR1A mutations in endocrine tissues.
Regulation of Phospholipase D and Secretion in Mast Cells by Protein Kinase A and Other Protein Kinases
Authors: CHOI WAHN SOO , CHAHDI AHMED , KIM YOUNG MI , FRAUNDORFER PAUL F. , BEAVEN MICHAEL A.
Published: July 2009

Abstract:Functions attributed to phospholipase (PL) D include the regulation of intracellular trafficking of Golgi‐derived vesicles and secretion of granules from mast cells. We have reported that activation of PLD and secretion in a rat mast cell (RBL‐2H3) line is substantially enhanced by cholera toxin, a known activator of protein kinase (PK) A. Here we review the evidence that (1) the synergistic interactions of cholera toxin and other pharmacological agents on mast cell secretion are attributable to the synergistic activation of PLD via PKA, CaM kinase II, and PKC and (2) both PLD1 and PLD2 participate in this process. For example, treatment with cholera toxin, thapsigargin, and phorbol 12‐myristate 13‐acetate (which activate PKA, CaM kinase II, and PKC, respectively) exhibit synergy in the stimulation of both PLD and secretion. These kinases and PLD are likely confined to membrane components, as similar synergistic interactions could be demonstrated in permeabilized cells. The regulation of PLD and secretion by these kinases is also apparent from studies of inhibitors of PKA and other kinases. Also, by overexpression of either PLD1 or PLD2 it is apparent that both isoforms respond to the same stimuli as endogenous PLD, although PLD1 is largely associated with secretory granules and PLD2 with plasma membrane. The studies reveal interesting differences in the regulation of the translocation of granules (regulated by PKA) and the fusion of these granules with the plasma membrane (regulated by Ca2+and PKC). The pathological/physiological implications of the regulation of PLD by PKA require further evaluation in other cell systems.
Relating National Veterinary Services to the Country's Livestock Industry
Authors: WINDSOR ROGER S.
Published: July 2009

Abstract: At the end of WW II, the British Government of the time decided that it was essential for Britain to become self‐sufficient in food. In consequence there was a large investment in services to agriculture and in particular many new veterinary investigation centers were opened to help farmers produce more animal products. The upsurge in world trade led the Government of Mrs. Thatcher to decide that livestock was just another commodity and so there has been a massive scaling down of money available to assist the livestock farmer. For Botswana the livestock industry is vital to the well‐being of the people and successive Governments have continued to invest in veterinary services. As a consequence, Botswana has one of the best and most efficient Veterinary Services in Africa. By contrast, the livestock industry in Perú has an insignificant effect on the gross national product. The fiber exports from camelids are a small international market, while the dairy industry is unable to provide sufficient milk for the nation. Partly as a result of this, the Peruvian Government invests very little in the livestock industry or the veterinary services that support it. Vietnam is in a transitional stage: there is a large but as yet unorganized livestock industry with a mass of smallholder farmers. The Government has made a large investment in people in the Department of Animal Health but without a concomitant investment in equipment and training. If the industry is to develop, it will require much more investment from the government. These countries will be discussed in more detail and an attempt will be made to show how by relating the services to the livestock industry, governments can improve services and at the same time cut the costs.
PCR and Molecular Detection for Differentiating Vibrio Species
Authors: SPARAGANO O.A. E. , ROBERTSON P.A. W. , PURDOM I. , McINNES J. , LI Y. , YU D.‐H. , DU Z.‐J. , XU H.‐S. , AUSTIN B.
Published: July 2009

Abstract: Vibriosis is an economically important disease of fish, marine invertebrates (particularly penaeid shrimps), and large marine mammals and is responsible for high mortality rates in aquaculture worldwide. Some Vibrio species are also responsible for zoonoses, whereas others are relatively nonpathogenic. Using 16S‐ and 23S‐based PCR reactions, we obtained species‐specific patterns and a 470‐bp band, respectively. DNA sequences obtained on the 23S rRNA gene allowed us to identify species‐specific probes for Vibrio parahaemolyticus, V. alginolyticus, V. anguillarum and for a cluster of taxonomically related species: V. carchariae/harveyi/campbelii. A phylogenetic tree based on the 23S sequences confirmed previous results obtained by Western blotting.
Early Diagnosis of Johne's Disease in the American Bison by Monoclonal Antibodies Directed against Antigen 85
Authors: LEID JEFF G. , HUNTER DAVID , SPEER C. A.
Published: July 2009

Abstract: Several monoclonal antibodies derived from hybridomas from mice that had been immunized with recombinant Mycobacterium bovis antigen 85 (Ag85) were tested for reactivity against antigen 85 (Ag85) from M. bovis and against sera from 100 bison inoculated with M. paratuberculosis and from 100 control bison from a disease‐free herd. Monoclonal antibodies mAb85.1, mAb85.44.1, mAb85.44.9, and mAb85.96 reacted against three or four 30‐33‐kDa bands of the Ag85 complex of M. bovis. Importantly, these mAbs also reacted with bands of similar molecular weight in the sera of bison inoculated with M. paratuberculosis. Additionally, when sera from 198 bison in four herds were reacted against mAb85.1 and mAb85.96, 26 bison reacted positively for the presence of Ag85 by either mAb or by both. These preliminary results indicate that monoclonal antibodies may eventually lead to a reliable diagnostic test for the early detection of M. paratuberculosis infections in ruminants as well as to a means for identifying contaminated dairy products.
Pan‐Mediterranean Comparison for the Molecular Detection of Theileria annulata
Authors: SPARAGANO OLIVIER A.E. , CARELLI GRAZIA , CECI LUIGI , SHKAP VARDA , MOLAD THEA , VITALE FABRIZIO , LORIA GUIDO R. , REALE STEFANO , CARACAPPA SANTO , BOUATTOUR ALI , ALMERIA SONIA , CASTELLA JOAQUIM , CORCHERO EDUARDO , HABELA MIGUEL
Published: July 2009

Abstract: Seven laboratories decided to compare their molecular diagnostic techniques to identify Mediterranean theileriosis caused by Theileria annulata. Each laboratory used either PCR or PCR and reverse line blot hybridization (RLB) to identify T. annulata. Five laboratories sent their own samples to laboratory 4 to be recoded and passed on to at least two other laboratories. A total of 120 blood samples were analyzed during this study, generating 540 results. Laboratory 1 sent only T. annulata‐infected samples (positive control batch), and all the laboratories testing this batch found 100% infection. Laboratory 2 sent only negative samples from a Mediterranean area where T. annulata was unknown, and two laboratories out of three found a few positive samples in these negative samples. For the remaining samples, detection performance was variable. Agreement between laboratories ranged from 21.4 to 91.3%. The overall mismatch between laboratories was around 30% by whatever technique used. This paper describes the methodological parameters that could explain the variation of results.
Reverse Line Blot Hybridization Used to Identify Hemoprotozoa in Minorcan Cattle
Authors: ALMERIA SONIA , CASTELLÀ JOAQUIM , FERRER DAVID , GUTIÉRREZ JUAN FRANCISCO , ESTRADA‐PEÑA AGUSTIN , SPARAGANO OLIVIER
Published: July 2009

Abstract: Piroplasmosis, a tick‐borne protozoal disease, is an important disease affecting domestic and wild animals. We performed PCR‐based reverse line blot hybridization (RLB) assays on blood samples obtained from 133 cattle exposed to ticks in field conditions in Minorca (Balearic Islands, Spain) in three different seasons. The oligonucleotides used were those for Theileria annulata, T. buffeli, T. taurotragi, T. velifera, Babesia bigemina, B. bovis, B. divergens, and B. major. The RLB technique allowed the simultaneous identification of T. annulata, T. buffeli, B. bigemina, and B. bovis as the piroplasms present in cattle in Minorca. Of the 133 animals, only 4 were not infected by any of the studied parasites. The results indicated endemic piroplasm infection in cattle in Minorca; especially important was the presence of T. annulata. The RLB was highly sensitive and allowed the simultaneous detection and identification of the Theileria and Babesia species in carrier cattle, which cannot be achieved by classical identification methods.
Construction and Evaluation of a Recombinant Foot‐and‐Mouth Disease Virus
Authors: VAN RENSBURG HESTER G. , MASON PETER W.
Published: July 2009

Abstract: The South African Territories (SAT) types of foot‐and‐mouth disease virus (FMDV) show marked genomic and antigenic variation throughout sub‐Saharan Africa. This variation is geographically linked and requires the use of custom‐made vaccines. Adaptation of field isolates as vaccine strains is cumbersome, time consuming, and expensive. As an alternative to the adaptation process, the construction of recombinant FMD viruses followed by the production of conventional, inactivated vaccines utilizing these viruses is proposed. The advantage of such a strategy would be the ability to manipulate the antigenicity of these viruses by substituting the antigenic coding regions (i.e., structural proteins) of a full‐length cDNA clone of a suitable strain. A chimeric cDNA clone between types A and SAT 2 was constructed by inserting the external capsid‐coding region of the vaccine strain ZIM/7/83/2 into the genetic backbone of the A12 cDNA clone. Preliminary evaluation of the recombinant FMD virus revealed a slower growth rate for the recombinant than the parental ZIM/7/83/2, although similar antigen yields could be obtained. The chimera was found to be thermally less stable than the parental strain, suggesting it to be an inferior strain for inactivated vaccine production.
Rapid Genotyping of Bacillus anthracis Strains by Real‐Time Polymerase Chain Reaction
Authors: PATRA GUY , WILLIAMS LEANNE E. , QI YUAN , ROSE SHARON , REDKAR RAJENDRA , DELVECCHIO VITO G.
Published: July 2009

Abstract: Rapid and accurate identification of Bacillus anthracis is critical for patient care as well as outbreak control. We have developed 3 separate PCR based assays using fluorescence resonance energy transfer (FRET) to detect the presence of pXO1, pXO2 plasmids and a chromosomal marker. A set of amplification primers and probes were used in each assay. The probes were ad jacently placed inside the primer sites and were 1‐bp apart. The upstream probe was labeled with fluorescein at the 3′ end, and the downstream probe had Cy5 attached at the 5′ end. The probes are included in the PCR reactions and hybridize with the PCR products as they are formed. Binding of probes to PCR products results in transfer of energy from fluorescein to Cy5, resulting in emission from Cy5. Increase in fluorescence, indicating amplification, was monitored in real time on a LightCycler™ LC24. Initial denaturation of target sequences was accomplished at 95°C for 1 min, followed by 28 cycles of denaturation at 95°C for 0 sec, annealing at 58°C for 15 sec, and elongation at 72°C for 5 sec. These assays are specific and can be performed on as little as 25 ng of total DNA or crude cell lysate a from fresh colony. It is thus possible to deter mine the genotype of B. anthracis strains in less than 1 hour.
IFN‐γ As an Indicator of Successful Immunization of Goats Vaccinated with a Killed Cowdria ruminantium Vaccine
Authors: ESTEVES I. , BENSAID A. , MARTINEZ D. , TOTTE P.
Published: July 2009

Abstract: Cowdria ruminantium‐induced production of IFN‐γ was measured by ELISA on a weekly basis during the course of vaccination with killed organisms emulsified in ISA50. Upon challenge, all (3/3) vaccinated animals that gave the lowest IFN‐γ response died of peracute cowdriosis. On the other hand, only one of three animals showing high IFN‐γ responses to vaccination died, but with a delay of 4 days in comparison with naïve controls. Thus, there seems to be a threshold level of IFN‐γ below which the probability for vaccinated animals to survive a lethal challenge is very low. During challenge, a much lower, but still physiologically meaningful production of IFN‐γ was detected using the 24‐hour whole blood assay on day 5 after infection in animals controlling the infection. In contrast, IFN‐γ production was absent or negligible in naïve and vaccinated animals that died within 8‐10 days after infection. Although these results need to be validated on a larger number of animals, they strongly suggest that IFN‐γ is a useful indicator of protective immunity in animals immunized with killed Cowdria.
Cowdria ruminantium Antigens of around 15 kDa Are Potent Inducers of IFN‐γ
Authors: GUNTER N. , ESTEVES I. , KANDASSAMY Y. , MARTINEZ D. , BENSAID A. , VAN KLEEF M. , DU PLESSIS D. , TOTTE P.
Published: July 2009

Abstract: Cellular responses induced in two Creole goats by vaccination with killed Cowdria ruminantium (Cowdria) were confirmed by IFN‐γ production and interleukin‐2 receptor (IL‐2R) expression. Both CD4+ and CD8+ but not WC1+ T cells showed a substantial increase in cell surface expression of IL‐2R molecules in response to whole Cowdria lysate. Cowdria (Welgevonden strain) proteins were fractionated using continuous‐flow electrophoresis and tested for their ability to induce IFN‐γ production by PBMC collected three weeks after the first inoculation and one week after the booster injection. Pooled fractions of around 15, 22, and 24 kDa were found to induce significant IFN‐γ production in both vaccinated animals on one of the two occasions. Antigens of around 15 kDa induced substantially higher IFN‐γ production than any other fractions in both animals. These pilot experiments pave the way towards the identification of proteins/genes that have potential for the development of a recombinant vaccine against heartwater.
Sequencing of a 15‐kb Ehrlichia ruminantium Clone and Evaluation of the cpg1 Open Reading Frame for Protection against Heartwater
Authors: LOUW ELMARIÉ , BRAYTON K. A. , COLLINS N. E. , PRETORIUS A. , VAN STRIJP F. , ALLSOPP B. A.
Published: July 2009

Abstract: A 1.2 kb polymorphic fragment from the Gardel isolate of Ehrlichia (formerly Cowdria) ruminantium was used to isolate a 15kb clone from the E. ruminantium Welgevonden LambdaGEM‐11 library. This clone, WL2EL1, was subcloned and sequenced. Eight open reading frames (ORFs) were identified. The ORF in WL2EL1 which contained the Welgevonden homologue of the 1.2 kb polymorphic fragment was designated Cowdria polymorphic gene 1 (cpg1). The cpg1 ORF was cloned into pCMViUB, a genetic vaccine vector. Mice and sheep were immunized with pCMViUB/cpg1 by intramuscular injection and gene gun inoculation. Although all of the immunized mice died, there was a trend for mice that received larger amounts of pCMViUB/cpg1 DNA to survive longer. Four out of five sheep immunized with the construct survived lethal challenge.
The Innate Resistance of Kenana Cattle to Tropical Theileriosis (Theileria annulata Infection) in the Sudan
Authors: BAKHEIT M. A. , LATIF A. A.
Published: July 2009

Abstract: A study was carried out to assess the innate resistance of the indigenous Kenana breed of cattle in the Sudan to tropical theileriosis, Theileria annulata infection of cattle. Nine susceptible Kenana calves were obtained from an area free from tropical theileriosis and the vector tick Hyalomma anatolicum anatolicum and were found negative to T. annulata antibodies in the indirect fluorescent antibody test. They were infected by inoculation of 1.0 mL T. annulata sporozoite stabilate. Three Friesian calves were also infected and served as susceptible controls. The percent of schizont parasitosis (Macroschizont Index, MSI) in the Kenana cattle was reduced by 70% compared to the Friesian calves. The percent of piroplasm parasitemia was also significantly lower in the Kenana calves. The rate of white blood cell reduction was significantly greater in the Friesian calves (P < 0.05). These differences were attributed to the high rate of schizont multiplication in the control cattle. Seventy‐eight percent (7/9) of the Kenana cattle recovered spontaneously, and only 22% required treatment compared to 100% mortality in the Friesian controls. These differences were attributed to the high rate of schizont multiplication in the control cattle and, on the other hand, ability of the Kenana cattle to limit the MSI, resulting in less severe damage to the lymphoid tissue during the acute phase of the disease.
Residual Effect of Antibabesial Drugs on the Live Redwater Blood Vaccines
Authors: COMBRINK M. P. , TROSKIE P. C. , DE WAAL D. T.
Published: July 2009

Abstract:It has been demonstrated that the attenuated organisms used in the unfrozen South AfricanBabesia bovisandB. bigemina(redwater) vaccines are susceptible for longer periods to the residual effect of the anti‐babesial drugs diminazene and imidocarb dipropionate than the virulent field strains. Reports of vaccine failures in some animals vaccinated with the frozen South African redwater vaccines after prophylactic treatment with imidocarb dipropionate have led us to reinvestigate the validity of the recommended prescribed waiting periods. Results indicated that waiting periods before administration of the frozenB. bovisandB. bigeminavaccines in animals that have been treated with diminazene at 3.5 mg/kg live weight, compare favorably with results initially obtained for the unfrozen vaccines at 4 and 8 weeks, respectively. However, the inhibitory effect of imidocarb dipropionate at 3.0 mg/kg live weight on the infectivity of both frozenB. bovisandB. bigeminavaccines is longer than previously anticipated and necessitated changing the minimum waiting periods before administration of these vaccines from 8 to 12 weeks and 16 to 24 weeks, respectively.