bluuebunny/crossref_metadata_2025 huggingface dataset:
Role of the mTOR Pathway in Normal and Tumoral Adrenal Cells
Authors: De Martino M.C. , van Koetsveld P.M. , Pivonello R. , Hofland L.J.
Published: September 2010

The mammalian target of rapamycin (mTOR) is a kinase of the phosphoinositide 3-kinase (PI3Ks)/protein kinase B (PKB or AKT) signaling pathway, which is one of the most important intracellular mediators of the activity of growth factors receptors, including vascular endothelial growth factor (VEGF) and insulin-like growth factors (IGFs). Dysregulation of the mTOR pathway has been found in many human tumors. Therefore, the mTOR pathway is considered as a target for antineoplastic therapy in several malignancies. Presently, the role and functions of mTOR and its signaling pathway in the normal and pathological adrenal gland has not been clarified yet. However, many growth factors and growth factor receptors, which are considered to play a role in the pathogenesis of adrenal tumors, can at least in part exert their effects through the activation of PI3K/AKT/mTOR pathway. Dysregulation of AKT has been reported in adrenocortical carcinomas and adrenomedullary tumors, named pheochromocytomas. Adrenocortical carcinomas and malignant pheochromocytomas are aggressive tumors with poor prognosis and scant treatment options. Therefore, new treatment options are warranted for these malignancies. On the basis of the current knowledge, mTOR could play a role in the pathogenesis of both adrenocortical carcinomas and pheochromocytomas. Moreover, mTOR inhibitors, interfering with the activation of several mitogenic and angiogenic factors, could be considered as a novel treatment opportunity for the management of malignant adrenal tumors.
Role of Dopamine Receptors in Normal and Tumoral Pituitary Corticotropic Cells and Adrenal Cells
Authors: Boschetti Mara , Gatto Federico , Arvigo Marica , Esposito Daniela , Rebora Alberto , Talco Miryam , Albertelli Manuela , Nazzari Elena , Goglia Umberto , Minuto Francesco , Ferone Diego
Published: September 2010

The recent depiction of dopamine receptors (DRs) in tumors that cause Cushing’s syndrome (CS) has renewed the debate about the dopamine control on pituitary-adrenal axis, and opened interesting new perspectives for medical treatment of CS. The new insights arise from the recent accurate characterization of DR subtypes expression within tumors causing CS, the discovery of new mechanisms, such as the dimerization between DRs and other G-protein coupled receptors (CPCRs), including somatostatin receptors (SSTRs), and the recent availability of new agents targeting these receptor subtypes. Corticotropic adenomas express DR subtype 2 (D2R), together with different SSTR subtypes (ssts), in particular sst5. In vitro, activation of D2R inhibits ACTH release in the majority of cultures of corticotropic cells, whereas, in vivo, dopaminergic agents display an inhibitory effect on cortisol levels in a subset of patients with CS. In animal models the receptor profile can be deeply modulated in specific environmental conditions, that may resemble the different clinical phases of CS. The new insights about DRs and receptor-targeting drugs may offer different approaches for medical treatment of CS: combination therapies with different types of compounds, treatment with novel molecules (hybrid compounds) with a wider spectrum of activity, or even pretreatment manipulation of receptor profile. Finally, recent studies showed that D2R is also significantly expressed in ectopic ACTH-secreting tumors and in both normal and tumoral adrenal tissues. Dopamine-agonists may decrease cortisol levels in a number of these patients, strengthening the current (re)emerging interest in DRs as possible targets for medical treatment of CS.
Role of Somatostatin Receptors in Normal and Tumoral Pituitary Corticotropic Cells
Authors: Hofland Leo J. , Lamberts Steven W.J. , Feelders Richard A.
Published: September 2010

Normal and tumoral pituitary corticotropic cells express sst2 and sst5, of which sst5 is the predominantly expressed receptor subtype. Somatostatin (SS) inhibits pituitary adrenocorticotropin hormone (ACTH) secretion in vitro, but the sensitivity to SS is strongly regulated by glucocorticoids. In pathological conditions of a low endogenous cortisol level, i.e. in patients with adrenal insufficiency and in patients with Nelson’s syndrome, SS and sst2-preferring SS analogs (SSA), such as octreotide, are able to lower circulating ACTH and cortisol levels. On the other hand, sst2-preferring SSA seem not effective in lowering ACTH and cortisol levels in patients with untreated Cushing’s disease (CD), in which circulating cortisol levels are high. This is likely due to the downregulation of sst2 receptors by glucocorticoids. sst5 receptor expression is more resistant to the inhibitory effect of glucocorticoids. In recent years, novel sst subtype-selective and universal SSA have been developed. In particular, SSA with a high sst5-binding affinity are potent inhibitors of ACTH secretion by pituitary corticotropic adenoma cells. This knowledge has initiated clinical trials evaluating the efficacy of these novel SSA in patients with CD, with the aim to lower circulating ACTH and cortisol levels by targeting multiple ssts on the corticotropic adenoma cells. In this minireview, the effects of SS in the regulation of normal and tumoral ACTH secretion, the role of sst subtypes involved herein, as well as the potentials of novel SSA in the treatment of patients with recurrent or persisting CD are discussed.
Epidemiology of Cushing’s Syndrome
Authors: Steffensen Charlotte , Bak Ann Mosegaard , Zøylner Rubeck Kristine , Jørgensen Jens Otto Lunde
Published: September 2010

Overt Cushing’s syndrome is a rare disorder with an annual incidence of 2–3/million of which benign adrenal adenomas account for 0.6/million. The female:male ratio is 3:1. Preliminary data indicate a high proportion of subclinical Cushing’s syndrome in certain risk populations such as patients with type 2 diabetes or osteoporosis. The clinical implications of these observations are presently unclear. Surgery remains first line treatment for overt disease and initial cure or remission is obtained in 65–85% of patients with Cushing’s disease. Late recurrences, however, occur in up to 20% and the risk does not seem to plateau even after 20 years of follow-up. A 2- to 3-fold increase in mortality is observed in most studies, and this excess mortality seems confined to patients in whom initial cure was not obtained. Cushing’s syndrome continues to pose diagnostic and therapeutic challenges and life-long follow-up is mandatory.
Skeletal Diseases in Cushing’s Syndrome: Osteoporosis versus Arthropathy
Authors: Kaltsas Gregory , Makras Polyzois
Published: September 2010

Structural and functional impairment of the skeletal system remains an important cause of morbidity and disability in patients with Cushing’s syndrome (CS). Glucocorticoid (GC) excess inhibits bone formation and calcium absorption from the gut, increases bone resorption, and alters the secretion of gonadotropin and growth hormones, cytokines and growth factors influencing bone. Both overt and subtle endogenous hypercortisolism affect bone, leading to vertebral fractures in up to 70% of patients. Fracture risk is related to age at onset, duration and severity of the disease and individual susceptibility to GCs that is genetically determined. Bone mineral density (BMD) measurement at the lumbar spine should be performed as a screening test in all patients with CS due to the preferential loss of trabecular bone induced by GCs. The higher risk of fractures at comparable BMD values with controls suggests that bone quality features, not assessed by routine BMD approaches, are also important and should be addressed when indicated applying specific radiological means. Successful treatment of GC excess is associated with improvement in bone mass which, although delayed and often incomplete, reduces the risk of osteoporotic fractures. Bisphosphonates can induce a more rapid improvement in BMD than cortisol normalization alone and can be used in patients with increased risks for further fractures and/or persistent hypercortisolemia to prevent further bone loss. Anabolic agents have not as yet been systemically used. Avascular necrosis, mainly of the femoral neck, and growth arrest in children are the most common skeletal disorders unrelated to osteoporosis encountered in patients with endogenous hypercortisolism.
Pituitary Radiotherapy for Cushing’s Disease
Authors: Losa Marco , Picozzi Piero , Redaelli Maria Grazia , Laurenzi Andrea , Mortini Pietro
Published: September 2010

Background: The treatment of choice for Cushing’s disease is pituitary surgery. Second-line treatments include repeat pituitary surgery, radiation therapy, medical therapy, and bilateral adrenalectomy. The most used modalities to irradiate patients with Cushing’s disease include fractionated radiotherapy and single-dose Gamma Knife. We aim to review the efficacy and safety of radiotherapy in patients with persistent or recurring Cushing’s disease. Results: Remission of Cushing’s disease after radiotherapy ranges from 42 to 83%. There seems to be no clear difference according to the technique of radiation used. Most patients experience remission of disease within 3 years from treatment, with only few cases reaching normal cortisol secretion after a longer follow-up. Control of tumor growth varies from 93 to 100%. Severe side effects of radiotherapy, such as optic neuropathy and radionecrosis, are uncommon. New-onset hypopituitarism is the most frequent side effect of radiation, occurring in 30–50% of patients treated by fractionated radiotherapy while it has been reported in 11–22% of patients after Gamma Knife. Conclusion: Radiotherapy is an effective second-line treatment in patients with Cushing’s disease not cured by surgery. Consideration of the advantages and disadvantages of radiotherapy in comparison with other therapeutic options should always be carried out in the single patient before deciding the second-line therapeutic strategy for persisting or recurring Cushing’s disease.
Hypertension in Cushing’s Syndrome: From Pathogenesis to Treatment
Authors: Cicala Maria Verena , Mantero Franco
Published: September 2010

Hypertension is one of the most distinguishing features of endogenous Cushing’s syndrome (CS), as it is present in about 80% of adult patients whereas in children its prevalence is about 47%. Hypertension in CS is significantly correlated with the duration of hypercortisolism and results from the interplay between several pathophysiological mechanisms regulating plasma volume, peripheral vascular resistance and cardiac output, all of which are increased in this state. Glucocorticoids cause hypertension through several mechanisms: their intrinsic mineralocorticoid activity; through activation of the renin-angiotensin system; by enhancement of vasoactive substances, and by causing suppression of the vasodilatory systems. In addition, glucocorticoids may exert some hypertensive effects on cardiovascular regulation through the CNS via both glucocorticoid and mineralocorticoid receptors. Hypertension in CS usually resolves with surgical removal of the tumor, but some patients require pharmacological antihypertensive treatment both pre- and postoperatively. Thiazides and furosemide should be avoided, while adrenergic blockade and calcium channel antagonists are usually ineffective. Mineralocorticoid receptor antagonists, Ang II blockers and ACE inhibitors are good anti-hypertensive options; PPAR-γ agonists may help in many aspects of the insulin resistance syndrome. The relatively selective glucocorticoid receptor antagonist Mifepristone (RU 486) could reduce blood pressure in patients with CS. Neuromodulatory agents such as the serotonin inhibitors cyproheptadine and ritanserin, valproid acid, dopamine agonists, somatostatin analogs may occasionally be effective, as well as drugs acting directly at the adrenal levels, such as Ketoconazole and aminoglutetimide or even opDDD. Treating hypertension in CS remains a difficult task and a big challenge, in order to decrease the morbidity and mortality associated with the disease.
Neuropsychiatric Disorders in Cushing’s Syndrome
Authors: Pereira Alberto M. , Tiemensma Jitske , Romijn Johannes A.
Published: September 2010

Glucocorticoids are crucial in the initiation and consolidation of the stress response. Patients with active Cushing’s syndrome (CS) are exposed to excessive endogenous glucocorticoid levels. In these patients, psychopathology is often being observed. The most common co-morbid disorder is major depression, but to a lesser extent mania and anxiety disorders have also been reported. A severe clinical presentation of CS often also includes depression. Reduction of glucocorticoid synthesis or action, either with metyrapone, ketoconazole, or mifepristone, rather than treatment with antidepressant drugs, is generally successful in relieving depressive symptoms, as well as other disabling symptoms. Following successful surgical treatment of hypercortisolism, both physical and psychiatric signs and symptoms improve substantially. However, it appears that patients do not completely return to their premorbid level of functioning and persistent impairment of quality of life and cognitive function has been reported despite long-term cure. At present, it is not clear whether, and to which extent, psychopathology still affects general well-being after long-term cure of CS.
Pathophysiology of Diabetes Mellitus in Cushing’s Syndrome
Authors: Pivonello Rosario , De Leo Monica , Vitale Pasquale , Cozzolino Alessia , Simeoli Chiara , De Martino Maria Cristina , Lombardi Gaetano , Colao Annamaria
Published: September 2010

Cushing’s syndrome is commonly complicated with an impairment of glucose metabolism, which is often clinically manifested as diabetes mellitus. The development of diabetes mellitus in Cushing’s syndrome is both a direct and indirect consequence of glucocorticoid excess. Indeed, glucocorticoid excess induces a stimulation of gluconeogenesis in the liver as well as an inhibition of insulin sensitivity both in the liver and in the skeletal muscles, which represent the most important sites responsible for glucose metabolism. In particular, glucocorticoid excess stimulates the expression of several key enzymes involved in the process of gluconeogenesis, with a consequent increase of glucose production, and induces an impairment of insulin sensitivity either directly by interfering with the insulin receptor signaling pathway or indirectly, through the stimulation of lipolysis and proteolysis and the consequent increase of fatty acids and amino acids, which contribute to the development of insulin resistance. Moreover, the peculiar distribution of adipose tissue throughout the body, with the predominance of visceral adipose tissue, significantly contributes to the worsening of insulin resistance and the development of a metabolic syndrome, which participates in the occurrence and maintenance of the impairment of glucose tolerance. Finally, glucocorticoid excess is able to impair insulin secretion as well as act at the level of the pancreatic beta cells, where it inhibits different steps of the insulin secretion process. This phenomenon is probably responsible for the passage from an impairment of glucose tolerance to an overt diabetes mellitus in susceptible patients with Cushing’s syndrome.
Antigenic Heterogeneity of the NS3 Proteins in Hepatitis C Virus Genotypes 1 and 6
Authors: Qiao Weizhen , Xie Ping , Cai Bing , Li Lingling
Published: September 2010

Objective: To investigate the effect of sequence variability between different genotypes of hepatitis C virus (HCV) on the antigenic properties of the NS3 proteins, and to evaluate the significance of the proteins in the serological diagnosis. Methods: The recombinant expression plasmids of pET/NS3-1 and pET/NS3-6 were constructed with the NS3 region fragments from HCV genotypes 1 and 6, respectively. The antigenic reactivity of the recombinant NS3 proteins was determined. The NS3 nucleotide sequences were aligned with 6 HCV full-length strains representing different genotypes. Results: The two recombinant NS3 proteins were generated and enzyme immunoassay tests were developed. The positive rates of anti-NS3 were 61.2 and 58.8% by using the NS3 antigens from HCV genotypes 1 and 6, respectively. This difference had no statistical significance (p > 0.05). However, it was interesting that 16 samples gave a discordant result for different genotype NS3 antigens.The two partial NS3 sequences shared approximately 63.9–79.9% identity with the different genotype strains. Conclusion: The primary structure of the HCV NS3 region is highly variable. Thus, sequence variability has a profound effect on the antigenic properties of the NS3 regions, and the antigenic differences should be taken into account for the development of more effective diagnostic tests.
The Importance of Nutrition in the Prevention of Ocular Disease with Special Reference to Cataract
Authors: Agte Vaishali , Tarwadi Kirtan
Published: September 2010

Background: The lens is the ocular structure most susceptible to oxidative damage. Antioxidants, micronutrients and phytochemicals have been extensively studied for their possible effects to prevent or delay the progression of various eye diseases. Objectives: A brief overview of the updated literature on the role of antioxidants and micronutrients in the prevention and treatment of ocular diseases is to be presented with an emphasis on cataract. Data Sources: PubMed search and individual papers from journals. Data Synthesis: The review discusses linkages of various micronutrients and antioxidants as well as oxidative stress with cataract. Dietary interventions as strategy for prevention of cataract and other ocular disorders are also reviewed. Conclusions: Consumption of food-based antioxidants like β-carotene lutein and zeaxanthin seem to be useful for the treatment of macular degeneration and cataracts. Supplements of vitamin A, vitamin C, vitamin E and zinc may prevent advanced age-related macular degeneration only in high-risk individuals.
Mechanisms of Retinal Ganglion Cell Injury in Aging and Glaucoma
Authors: Chrysostomou Vicki , Trounce Ian A. , Crowston Jonathan G.
Published: September 2010

Aging is the greatest risk factor for glaucoma, implying that intrinsic age-related changes to retinal ganglion cells, their supporting tissue or both make retinal ganglion cells susceptible to injury. Changes to the ocular vasculature, connective tissue of the optic nerve head and mitochondria, which have been documented with advancing age and shown to be exacerbated in glaucoma, may predispose to glaucomatous injury. When considering such age-related changes, it is difficult to separate pathological change from physiological change, and cause from consequence. The insults that predispose aged retinal ganglion cells to injury are likely to be varied and multiple; therefore, it may be more relevant to identify and treat common mechanisms that predispose to retinal ganglion cell failure and/or death. We suggest that mitochondrial dysfunction, as either a cause or consequence of injury, renders retinal ganglion cells sensitive to degeneration. Therapeutic approaches that target mitochondria and promote energy production may provide a general means of protecting aged retinal ganglion cells from degeneration, regardless of the etiology.
Oxidative Damage and the Prevention of Age-Related Cataracts
Authors: Beebe David C. , Holekamp Nancy M. , Shui Ying-Bo
Published: September 2010

Purpose: Cataracts are often considered to be an unavoidable consequence of aging. Oxidative damage is a major cause or consequence of cortical and nuclear cataracts, the most common types of age-related cataracts. Methods: In this review, we consider the different risk factors, natural history and etiology of each of the 3 major types of age-related cataract, as well as the potential sources of oxidative injury to the lens and the mechanisms that protect against these insults. The evidence linking different oxidative stresses to the different types of cataracts is critically evaluated. Results: We conclude from this analysis that the evidence for a causal role of oxidation is strong for nuclear, but substantially lower for cortical and posterior subcapsular cataracts. The preponderance of evidence suggests that exposure to increased levels of molecular oxygen accelerates the age-related opacification of the lens nucleus, leading to nuclear cataract. Factors in the eye that maintain low oxygen partial pressure around the lens are, therefore, important in protecting the lens from nuclear cataract. Conclusions: Maintaining or restoring the low oxygen partial pressure around that lens should decrease or prevent nuclear cataracts.
Retinal Aging and Sirtuins
Authors: Ozawa Yoko , Kubota Shunsuke , Narimatsu Toshio , Yuki Kenya , Koto Takashi , Sasaki Mariko , Tsubota Kazuo
Published: September 2010

The process of aging involves the accumulating changes in the microenvironment that lead to cell senescence or apoptosis, and subsequent tissue or organ dysfunction. Multiple extrinsic and intrinsic events that cause DNA instability are associated with aging. Cells containing unstable DNA are biologically vulnerable, and if the DNA damage is too great for the cell to repair, it becomes senescent or dies by apoptosis. Thus, the cell’s capacity to repair its DNA determines the progress of aging, at least in part. Here, we focus on the sirtuins, the mammalian homologs of the yeast life-span-extending molecule, Sir2. Among the sirtuin family proteins in mammals, the one most similar to yeast Sir2 is SIRT1, which is involved in multiple pathways, including the repair of DNA double-strand breaks. Although the role of SIRT1 in mammalian longevity is not clear, it is expressed throughout the retina, where it may suppress aging. In fact, a mutant mouse model of retinal degeneration shows an abnormal subcellular localization of SIRT1 protein and accelerated retinal cell apoptosis. Further analyses are required to elucidate the mechanism of DNA damage and repair, including the contributions of the sirtuins, in the aged or diseased retinas, which will help us understand the mechanisms of retinal aging.
Repeat Traumatic Brain Injury in the Juvenile Rat Is Associated with Increased Axonal Injury and Cognitive Impairments
Authors: Prins M.L. , Hales A. , Reger M. , Giza C.C. , Hovda D.A.
Published: September 2010

Among the enormous population of head-injured children and young adults are a growing subpopulation who experience repeat traumatic brain injury (RTBI). The most common cause of RTBI in this age group is sports-related concussions, and athletes who have experienced a head injury are at greater risk for subsequent TBI, with consequent long-term cognitive dysfunction. While several animal models have been proposed to study RTBI, they have been shown to either produce injuries too severe, were conducted in adults, involved craniotomy, or failed to show behavioral deficits. A closed head injury model for postnatal day 35 rats was established, and single and repeat TBI (1-day interval) were examined histologically for axonal injury and behaviorally by the novel object recognition (NOR) task. The results from the current study demonstrate that an experimental closed head injury in the rodent with low mortality rates and absence of gross pathology can produce measurable cognitive deficits in a juvenile age group. The introduction of a second injury 24 h after the first impact resulted in increased axonal injury, astrocytic reactivity and increased memory impairment in the NOR task. The histological evidence demonstrates the potential usefulness of this RTBI model for studying the impact and time course of RTBI as it relates to the pediatric and young adult population. This study marks the first critical step in experimentally addressing the consequences of concussions and the cumulative effects of RTBI in the developing brain.
Corpus Callosum and Inferior Forebrain White Matter Microstructure Are Related to Functional Outcome from Raised Intracranial Pressure in Child Traumatic Brain Injury
Authors: Tasker Robert C. , Gunn Westland Amber , White Deborah K. , Williams Guy B.
Published: September 2010

In severe paediatric traumatic brain injury (TBI), a common focus of treatment is raised intracranial pressure (ICP). We have previously reported frontal cerebral vulnerability with executive deficits from raised ICP in paediatric TBI. Now, using diffusion tensor imaging (DTI) in a different population, we have examined fractional anisotropy (FA), and mean, axial and radial diffusivity (MD, AD, RD) in 4 regions of the corpus callosum (CC) and in both inferior frontal regions. Our aim was to examine during the chronic phase of TBI whether the CC cross-sectional area correlated with regional DTI metrics of white matter microstructure, with global outcome ratings of function (Functional Independence Measure and Multiattribute Health Status Classification) and with performance in the Rey-Osterrieth Complex Figure (ROCF) test. We examined 33 paediatric TBI cases who were followed, on average, 4.9 years after severe injury. All cases had received mechanical ventilation during their acute treatment and, a priori, they were assigned to a non-ICP or a raised ICP group. Twenty-two participants had mainly right-sided injury at the time of acute ictus. The findings confirm that severe TBI in childhood, complicated by intracranial hypertension, results in CC vulnerability. In the chronic phase of recovery, it is reduced in the cross-sectional area, it is more compact and thinned, and the anterior region is disproportionately small. Late after raised ICP, we have also found that individuals exhibit regional microstructural abnormality with combined reduced FA and increased MD, AD and RD. Smaller size and such microstructural changes in the anterior CC were associated with similar right-sided (rather than left-sided) frontal microstructural changes in the ICP group. Taken together, this evidence points to an interaction between raised ICP-related brain tissue perturbation and focal frontal extracallosal injury, leading to anterior CC regional vulnerability, most likely wallerian degeneration. At long-term follow-up, this lack of white matter integrity in the anterior CC is correlated with functional outcome, particularly in aspects of social interaction and the copy component of the ROCF test, which suggests that the CC-to-forebrain function warrants further study in chronic TBI.
Polihexanide: A Safe and Highly Effective Biocide
Authors: Kaehn K.
Published: September 2010

Polihexanide is a broad-spectrum antiseptic with excellent tolerance and a low-risk profile. The physicochemical action on the bacterial envelope prevents or impedes the development of resistant bacterial strains. Thus, polihexanide is particularly suitable and useful in the struggle against multidrug-resistant bacteria. The ecological database is still incomplete. There is some evidence that biodegradation requires adsorption to inert surfaces and that only a small number of bacterial species are capable of utilizing polihexanide.
Review on the Efficacy, Safety and Clinical Applications of Polihexanide, a Modern Wound Antiseptic
Authors: Hübner N.-O. , Kramer A.
Published: September 2010

Infected wounds are still one of the great challenges in medicine. In the last decade, it has become increasingly clear that antimicrobial chemotherapy is limited by the spread of antimicrobial resistance. Fortunately, new, highly effective antiseptic substances with a broad antimicrobial spectrum are available, so local treatment is expected to get increasingly more important in wound therapy. This paper reviews the antiseptic agent polihexanide (polyhexamethylene biguanide, PHMB), one of the most promising substances available today, from a clinical point of view, focusing on efficacy, safety and clinical applications.
Cardiovascular Disease in Cushing’s Syndrome: Heart versus Vasculature
Authors: De Leo Monica , Pivonello Rosario , Auriemma Renata S. , Cozzolino Alessia , Vitale Pasquale , Simeoli Chiara , De Martino Maria Cristina , Lombardi Gaetano , Colao Annamaria
Published: September 2010

Cushing’s syndrome (CS) causes metabolic abnormalities that determine an increased cardiovascular risk not only during the active phase of the disease but also for a long time after cure. Cardiovascular complications, such as premature atherosclerosis, coronary artery disease, heart failure, and stroke, in patients with CS cause a mortality rate higher than that observed in a normal population. The increased cardiovascular risk is mainly due to metabolic complications, such as metabolic syndrome, but also to vascular and cardiac alterations such as atherosclerosis and cardiac structural and functional changes. In the clinical management of patients with CS the focus should be on identifying the global cardiovascular risk and the aim should be to control not only hypertension but also other correlated risk factors, such as obesity, glucose intolerance, insulin resistance, dyslipidemia, endothelial dysfunction and the prothrombotic state. Considering that remission from hypercortisolism is often difficult to achieve and that the cardiovascular risk can persist even during disease remission, care and control of all cardiovascular risk factors should be one of the primary goals during the follow-up of these patients.
Quantitative Determination of Hepatitis C Core Antigen in Therapy Monitoring for Chronic Hepatitis C
Authors: Moscato Giovanna A. , Giannelli Gianluigi , Grandi Barbara , Pieri Daniela , Marsi Ombretta , Guarducci Isabella , Batini Irene , Altomare Emanuele , Antonaci Salvatore , Capria Alfonso , Pellegrini Giovanni , Sacco Rodolfo
Published: September 2010

The correlation and kinetics of hepatitis C virus (HCV) RNA and HCV core antigen levels in chronic hepatitis C patients treated with pegylated interferon + ribavirin were evaluated in order to envision a combined use of the two assays in therapy monitoring. HCV core antigen levels by a chemiluminescent immunoassay (Abbott ARCHITECT) and HCV-RNA levels by branched DNA (bDNA) or real-time PCR have been evaluated on plasma specimens from 32 patients treated for chronic hepatitis C. An early virological response (undetectable levels of HCV-RNA 4 weeks after start of treatment) was found in 10/23 subjects (43.5%) followed up for 5 months or more. The response was linked to the HCV genotype (20% in genotype 1B vs. 61.5% in other genotypes; p < 0.05). HCV RNA and HCV antigen showed a good correlation (r = 0.814); HCV antigen was still detectable in 3 samples with undetectable (<615 IU/ml) RNA by bDNA, while no differences in clinical sensitivity were recorded in comparison with real-time PCR. These findings suggest that HCV-RNA and HCV antigen may be used at different time points in order to tailor therapy monitoring to individual needs.
Increased Neutralizing Antibody Response after Simultaneous Immunization with Leucogen and the Feline Leukemia Virus Transmembrane Protein
Authors: Langhammer Stefan , Fiebig Uwe , Kurth Reinhard , Denner Joachim
Published: September 2010

To develop improved vaccination strategies against feline leukemia virus (FeLV), rats were immunized with the transmembrane envelope protein p15E of FeLV alone or in combination with the commercial vaccine Leucogen® comprising the nonglycosylated FeLV surface envelope protein. Binding and neutralizing antibodies were induced in both groups and in the group immunized with Leucogen alone. Higher titers of antibodies neutralizing FeLV were induced by simultaneous immunization with Leucogen and p15E compared to the responses using Leucogen or p15E alone, suggesting that combination vaccines should be used in the future. Epitope mapping of p15E-specific antibodies induced by simultaneous immunization with Leucogen and p15E revealed the same pattern of response as obtained after immunization with p15E alone: one epitope was localized in the membrane-proximal external region (MPER) and the other in the fusion peptide-proximal region, and they are related to the epitopes detected after immunization with p15E of the porcine endogenous retrovirus and the koala retrovirus. The data indicate that these epitopes in the MPER are an effective target for neutralization and that antigens containing them may therefore prove to be a useful component of vaccines against retroviruses, including HIV-1.
Activation of T-Cell-Factor-Dependent Transcription by Kaposi’s Sarcoma-Associated Herpesvirus Replication Transactivation Activator
Authors: Cha Seho , Yoon Hee Jung , Kim Su-Jung , Seo Taegun
Published: September 2010

Objective: To elucidate the cellular function of Kaposi’s sarcoma-associated herpesvirus (KSHV) replication transactivation activator (RTA) at the transcriptional level. Methods: Transcriptional activation of T-cell-factor (TCF)-dependent genes by RTA was determined using the luciferase reporter assay. The specific regions of RTA required for the activation of TCF-dependent transcription and association with β-catenin were established. Results: RTA specifically activated TCF-dependent transcription in a dose-dependent manner, to an extent comparable to the activation achieved by latency-associated nuclear antigen. In contrast, other KSHV viral proteins investigated, such as basic leucine zipper and viral interferon regulatory factor 1, did not affect TCF-dependent transcription. The C-terminal region of RTA appeared to be necessary for transcription. However, RTA did not affect the β-catenin level or the subcellular localization thereof. Conclusion: Our results collectively demonstrate that KSHV RTA activates TCF-dependent transcription without involving the β-catenin pathway.
Factors That Influence Data Quality in Caries Experience Detection: A Multilevel Modeling Approach
Authors: Mutsvari T. , Lesaffre E. , García-Zattera M.J. , Diya L. , Declerck D.
Published: September 2010

Caries experience detection is prone to misclassification. For this reason, calibration exercises which aim at assessing and improving the scoring behavior of dental raters are organized. During a calibration exercise, a sample of children is examined by the benchmark scorer and the dental examiners. This produces a 2 × 2 contingency table with the true and possibly misclassified responses. The entries in this misclassification table allow to estimate the sensitivity and the specificity of the raters. However, in many dental studies, the uncertainty with which sensitivity and specificity are estimated is not expressed. Further, caries experience data have a hierarchical structure since the data are recorded for the surfaces nested in the teeth within the mouth. Therefore, it is important to report the uncertainty using confidence intervals and to take the clustering into account. Here we apply a Bayesian logistic multilevel model for estimating the sensitivity and specificity. The main goal of this research is to find the factors that influence the true scoring of caries experience accounting for the hierarchical structure in the data. In our analysis, we show that the dentition type and tooth or surface type affect the quality of caries experience detection.
Response Analysis of Stimulating Efficacy of Polihexanide in an in vitro Wound Model with Respiratory Ciliary Epithelial Cells
Authors: Roth C. , Beule A.G. , Kramer A. , Hosemann W. , Kohlmann T. , Scharf C.
Published: September 2010

In animal wound models, accelerated wound closure has been shown by use of polihexanide applied in antimicrobially effective concentrations. Additionally, an increased ATP production of keratinocytes in vitro induced by polihexanide was demonstrated and interpreted as a stimulatory effect on cell proliferation. Based on these results and the clinical reports on improved wound healing after introduction of polihexanide for preoperative antisepsis in the nasal cavity, polihexanide was tested in a wound model on respiratory ciliary epithelial cells allowing measurement of the healing process after artificial injury. 0.5 µg/ml polihexanide accelerated wound healing in terms of proliferation and migration significantly after an exposure time of 1 and 96 h. At a concentration of 1 µg/ml polihexanide, the stimulation of wound healing was significantly increased only after an exposure time of 96 h. This is the first study to demonstrate acceleration of wound healing in a standardized in vitro model using an epithelial cell line. Considering the present results and previous reports on the impact of polihexanide on wound healing, the conclusion is drawn that the positive effect of polihexanide on wound healing is a separate, dose-dependent effect independent of its antiseptic properties.
Blockade of Proteinase-Activated Receptor-4 Inhibits the Eosinophil Recruitment Induced by Eotaxin-1 in the Pleural Cavity of Mice
Authors: Braga A.D. , Miranda J.P. , Ferreira G.M. , Bilheiro R.P. , Duarte I.D. , Francischi J.N. , Klein A.
Published: September 2010

Objective and Design: Although proteinase-activated receptor (PAR)-4 has been implicated in inflammation, its role in regulating eosinophil recruitment in response to chemoattractants has not yet been demonstrated. To investigate the contribution of proteinases and PAR-4 activation to eosinophil migration in response to eotaxin-1 or leukotriene B4 (LTB4), the effects of aprotinin or PAR-4 antagonist trans-cinnamoyl-YPGKF-NH2 (tcY-NH2) on eosinophil migration induced by these chemoattractants were investigated. Methods: BALB/c mice were pretreated with aprotinin or tcY-NH2 (30 µg/mouse) prior to intrapleural injection of LTB4 or eotaxin-1 and the number of infiltrating eosinophils was determined 48 h later. Results: Aprotinin (1 mg/kg) inhibited eosinophil recruitment induced by eotaxin-1 (p < 0.01), but not that induced by LTB4. Moreover, tcY-NH2 treatment inhibited eosinophil recruitment in response to eotaxin-1 (p < 0.01 by ANOVA/Tukey post-test). Conclusion: These data suggest that aprotinin-inhibited proteinases participate in eosinophil migration induced by eotaxin-1 and that PAR-4 activation plays an important role in regulating this migration.
Inactivation of Macrophage Rab7 by Burkholderia cenocepacia
Authors: Huynh Kassidy K. , Plumb Jonathan D. , Downey Gregory P. , Valvano Miguel A. , Grinstein Sergio
Published: September 2010

Strains of the Burkholderia cepacia complex can survive within macrophages by arresting the maturation of phagocytic vacuoles. The bacteria preclude fusion of the phagosome with lysosomes by a process that is poorly understood. Using murine macrophages, we investigated the stage at which maturation is arrested and analyzed the underlying mechanism. Vacuoles containing B. cenocepacia strain J2315, an isolate of the transmissible ET12 clone, recruited Rab5 and synthesized phosphatidylinositol-3-phosphate, indicating progression to the early phagosomal stage. Despite the fact that the B. cenocepacia-containing vacuoles rarely fused with lysosomes, they could nevertheless acquire the late phagosomal markers CD63 and Rab7. Fluorescence recovery after photobleaching and use of a probe that detects Rab7-guanosine triphosphate indicated that activation of Rab7 was impaired by B. cenocepacia, accounting at least in part for the inability of the vacuole to merge with lysosomes. The Rab7 defect was not due to excessive cholesterol accumulation and was confined to the infected vacuoles. Jointly, these experiments indicate that B. cenocepacia express virulence factors capable of interfering with Rab7 function and thereby with membrane traffic.
Nuclease Expression by Staphylococcus aureus Facilitates Escape from Neutrophil Extracellular Traps
Authors: Berends Evelien T.M. , Horswill Alexander R. , Haste Nina M. , Monestier Marc , Nizet Victor , von Köckritz-Blickwede Maren
Published: September 2010

Neutrophils are key effectors of the host innate immune response against bacterial infection. Staphylococcus aureus is a preeminent human pathogen, with an ability to produce systemic infections even in previously healthy individuals, thereby reflecting a resistance to effective neutrophil clearance. The recent discovery of neutrophil extracellular traps (NETs) has opened a novel dimension in our understanding of how these specialized leukocytes kill pathogens. NETs consist of a nuclear DNA backbone associated with antimicrobial peptides, histones and proteases that provide a matrix to entrap and kill various microbes. Here, we used targeted mutagenesis to examine a potential role of S. aureus nuclease in NET degradation and virulence in a murine respiratory tract infection model. In vitro assays using fluorescence microscopy showed the isogenic nuclease-deficient (nuc-deficient) mutant to be significantly impaired in its ability to degrade NETs compared with the wild-type parent strain USA 300 LAC. Consequently, the nuc-deficient mutant strain was significantly more susceptible to extracellular killing by activated neutrophils. Moreover, S. aureus nuclease production was associated with delayed bacterial clearance in the lung and increased mortality after intranasal infection. In conclusion, this study shows that S. aureus nuclease promotes resistance against NET-mediated antimicrobial activity of neutrophils and contributes to disease pathogenesis in vivo.
Association between the Severity of Cerebral Small Vessel Disease, Pulsatility of Cerebral Arteries, and Brachial Ankle Pulse Wave Velocity in Patients with Lacunar Infarction
Authors: Kim Dae-Hyun , Choi Jae-Hyeong , Moon Jung-Soo , Kim Hyun-Jung , Cha Jae-Kwan
Published: September 2010

Background/Aims: Stiff arteries cause high pulse pressure and flow to be transmitted to distal organs during systole, damaging the cerebral microvasculatures. We investigated the association between the severity of cerebral small vessel disease (CSVD), the pulsatility of cerebral arteries, and the stiffness of large arteries in patients with lacunar infarction (LI). Methods: Subjects included 120 patients with LIs, who underwent MRI, transcranial Doppler ultrasonography, and brachial ankle pulse wave velocity (BaPWV). We compared the severity of CSVD, the pulsatile index (PI) of both middle cerebral arteries (MCAs), and BaPWV. Results: Patients with higher BaPWV were more likely to have multiple LIs and more severe white matter lesions, and tended to have higher age, systolic blood pressure, pulse pressure, heart rate, and the PI of MCAs. The PI of MCAs significantly correlated with BaPWV (with PI: r = 0.441, p < 0.001) and age (with PI: r = 0.538, p < 0.001). Multiple linear regression analysis indicated that aging (β = 0.008, p < 0.001) and BaPWV (β = 0.001, p = 0.007) were independent determinants of the PI of MCAs after adjustment for sex, pulse pressure, hypertension, and diabetes. Conclusion: These findings suggest that systemic arterial stiffness may be associated with the severity of CSVD and pulsatility of cerebral arteries.
Does ambulation modify venous thromboembolism risk in acutely ill medical patients?
Authors: Girard Frederick , Samama Meyer , Amin Alpesh
Published: September 2010

In the US, ambulatory status is often a criterion for stopping prophylaxis for venous thromboembolism (VTE). In an analysis of the prophylaxis in MEDical patients with ENOXaparin (MEDENOX) trial, ambulatory status was assessed as outcome and patients grouped accordingly for further analysis. Rates of VTE and bleeding were evaluated. Using multivariate logistic regression, the relationships between thromboprophylaxis, VTE risk, and ambulatory status were assessed. Ambulatory status was reached in 607/1,084 patients, in a mean time of 4.4 days. Thromboprophylaxis was provided for 7.3 and 7.7 days in the ambulatory and non-ambulatory groups. Although VTE rates were lower in ambulatory patients, enoxaparin 40 mg once daily significantly reduced the risk of VTE vs. placebo in ambulatory (3.3% vs. 10.6%; relative risk [RR] = 0.31; 95% confidence interval [CI], 0.13ﺹ0.78; p=0.008) and non-ambulatory patients (9.0% vs. 19.7%; RR = 0.46; 95% CI, 0.23ﺹ0.91; p=0.02). Major bleeding was not significantly different between enoxaparin and placebo in either group. By multivariate regression analysis, VTE risk in ambulatory patients was lower with enoxaparin vs. placebo (odds ratio [OR] = 0.28; 95% CI, 0.11ﺹ0.74; p=0.01), but higher in patients with a history of VTE (OR = 3.74; 95% CI, 1.59ﺹ8.84; p=0.003) or cancer (OR = 2.12; 95% CI, 1.00ﺹ4.48; p=0.049). Despite timely mobilisation, patients who become ambulatory are at VTE risk and experience a significant risk reduction with enoxaparin 40 mg. Therefore, it is essential that ambulatory patients receive recommended thromboprophylaxis.
Association between inflammatory biomarkers and platelet aggregation in patients under chronic clopidogrel treatment
Authors: Bernlochner Isabell , Steinhubl Steven , Braun Siegmund , Morath Tanja , Jaitner Juliane , Stegherr Julia , Mehilli Julinda , Beckerath Nicolas von , Schömig Albert , Kastrati Adnan , Sibbing Dirk
Published: September 2010

Inflammatory processes in the vessel wall are associated with progression of atherosclerosis and myocardial infarction. Both high levels of C-reactive protein (CRP) and high on-clopidogrel treatment platelet reactivity (HPR) have been linked to an increased risk of ischaemic events after percutaneous coronary intervention (PCI). The aim of this study was to explore the association between biomarker levels of inflammation and platelet reactivity. Stable patients (n=1,223) eligible for this study were under chronic antiplatelet treatment with aspirin and clopidogrel due to prior coronary stent placement. ADP-induced platelet aggregation (in AU*min) was measured on a Multiplate analyser. The primary outcome measure of this retrospective study was the ADP-induced platelet aggregation in patients with versus those without elevated CRP levels. Of the patients 15.5% (n=189) showed elevated CRP levels (≥5 mg/l). Platelet aggregation (median [interquartile range]) was significantly higher in patients with elevated CRP levels compared to patients with normal (<5 mg/l) CRP levels (305 [202–504] AU*min vs. 218 [144–384] AU*min; p<0.001). A multivariable linear regression model that adjusted for known predictors of HPR confirmed a significant independent association between elevated CRP levels and high ADP-induced platelet aggregation values (p=0.0002). Elevated WBC count and fibrinogen levels were also associated with higher platelet aggregation values (p<0.001 for both). In conclusion, elevated levels of CRP, WBC count and fibrinogen were significantly associated with high platelet reactivity in patients under chronic clopidogrel treatment. Whether a direct relation between platelets and inflammation exists, as well as the clinical impact of our results, warrants further investigations.