Cloning and characterization of rhesus IL-18 binding protein, a natural antagonist to IL-18.

IL-18 is a proinflammatory cytokine that is important for host defense, but is also involved in the pathogenesis of a number of disease processes, ranging from autoimmune disorders to atherosclerosis. IL-18 binding protein (IL-18BP) is a constitutively expressed glycoprotein that specifically neutralizes the effects of IL-18, resulting in decreased production of IFN-gamma and reduction in Th1 immune responses. In this study we cloned and sequenced a full-length cDNA of the rhesus IL-18BP (RhIL-18BP) from the spleen of rhesus macaques (Macaca mulatta) and compared its nucleotide and amino acid sequences to the functional murine and human IL-18BP orthologues. In addition, we fused RhIL-18BP to the Fc portion of human IgG1 to make recombinant RhIL-18BP x Fcgamma1 in order to facilitate its detection by Western blot analysis and determined the approximate molecular weight of RhIL-18BP x Fcgamma1 to be 66 kD. With this fusion protein, we showed that RhIL-18BP was functional and could significantly reduce murine IL-18 and LPS-induced IFN-gamma production by murine splenocytes. Furthermore, we demonstrated the expression of IL-18BP in atherosclerotic lesions in a rhesus model of atherosclerosis, underscoring the need to fully understand the role of this protein as a primary negative regulator of IL-18 in multiple disease processes.

Myocarditis in CD8-depleted SIV-infected rhesus macaques after short-term dual therapy with nucleoside and nucleotide reverse transcriptase inhibitors.

BACKGROUND: Although highly active antiretroviral therapy (HAART) has dramatically reduced the morbidity and mortality associated with HIV infection, a number of antiretroviral toxicities have been described, including myocardial toxicity resulting from the use of nucleotide and nucleoside reverse transcriptase inhibitors (NRTIs). Current treatment guidelines recommend the use of HAART regimens containing two NRTIs for initial therapy of HIV-1 positive individuals; however, potential cardiotoxicity resulting from treatment with multiple NRTIs has not been addressed. METHODOLOGY/PRINCIPAL FINDINGS: We examined myocardial tissue from twelve CD8 lymphocyte-depleted adult rhesus macaques, including eight animals infected with simian immunodeficiency virus, four of which received combined antiretroviral therapy (CART) consisting of two NRTIs [(9-R-2-Phosphonomethoxypropyl Adenine) (PMPA) and (+/-)-beta-2',3'-dideoxy-5-fluoro-3'-thiacytidine (RCV)] for 28 days. Multifocal infiltrates of mononuclear inflammatory cells were present in the myocardium of all macaques that received CART, but not untreated SIV-positive animals or SIV-negative controls. Macrophages were the predominant inflammatory cells within lesions, as shown by immunoreactivity for the macrophage markers Iba1 and CD68. Heart specimens from monkeys that received CART had significantly lower virus burdens than untreated animals (p<0.05), but significantly greater quantities of TNF-α mRNA than either SIV-positive untreated animals or uninfected controls (p<0.05). Interferon-γ (IFN-γ), IL-1ß and CXCL11 mRNA were upregulated in heart tissue from SIV-positive monkeys, independent of antiretroviral treatment, but CXCL9 mRNA was only upregulated in heart tissue from macaques that received CART. CONCLUSIONS/SIGNIFICANCE: These results suggest that short-term treatment with multiple NRTIs may be associated with myocarditis, and demonstrate that the CD8-depleted SIV-positive rhesus monkey is a useful model for studying the cardiotoxic effects of combined antiretroviral therapy in the setting of immunodeficiency virus infection.

Reduced genetic diversity in lymphoid and central nervous system tissues and selection-induced tissue-specific compartmentalization of neuropathogenic SIVsmmFGb during acute infection.

The simian lentivirus strain SIVsmmFGb is a viral swarm population inducing neuropathology in over 90% of infected pigtailed macaques and serves as a reliable model for HIV neuropathogenesis. However, little is understood about the genetic diversity of this virus, how said diversity influences the initial seeding of the central nervous system and lymph nodes, or whether the virus forms distinct genetic compartments between tissues during acute infection. In this study, we establish that our SIVsmmFGb stock virus contains four genetically distinct envelope V1 region groups, three distinct integrase groups, and two Nef groups. We demonstrate that initial central nervous system and lymph node seeding reduces envelope V1 and integrase genetic diversity but has a variable effect on Nef diversity. SIVsmmFGb envelope V1 region genes from the basal ganglia, cerebellum, and hippocampus form distinct genetic compartments from each other, the midfrontal cortex, and the lymph nodes. Basal ganglia, cerebellum, hippocampus, and midfrontal cortex-derived nef genes all form distinct genetic compartments from each other, as well as from the lymph nodes. We also find basal ganglia, hippocampus, and midfrontal cortex-derived integrase sequences forming distinct compartments from both of the lymph nodes and that the hippocampus and midfrontal cortex form separate compartments from the cerebellum, while the axillary and mesenteric lymph nodes compartmentalize separately from each other. Compartmentalization of the envelope V1 genes resulted from positive selection, and compartmentalization of the nef and integrase genes from negative selection. These results indicate restrictions on virus genetic diversity during initial tissue seeding in neuropathogenic SIV infection.

[Ambulatory cryosurgical restoration of benign cervical lesions]. / Ambulante kryochirurgische Sanierung gutartiger Portioveränderungen.

The area of ectopic epithelium and transformations zone of the uterine cervix was treated cryosurgically by liquid nitrogen in 252 patients. The influence of deep freeze lasted four minutes. Neither analgesia nor anaesthesia was necessary. After a single cryosurgery cure rate was 92 per cent, after a second application 98 per cent. Cryotherapy can be performed ambulatorily and is nowadays to be regarded as method of choice to restore benign cervical lesions.

[Bone scintigraphy following cryodevitalization of the mandible]. / Knochenszintigraphie nach Kryodevitalisation der Mandibula.

A tumor of the mandible can be destroyed successfully by cryogenic devitalisation. The revitalisation process was observed in 6 miniature swines by means of nuclear medical bone exploration. The bone metabolism decreased clearly postoperatively, and indeed laterally more than lingually. An increased blood flow and bone metabolism was recognized as an expression of the repair process at the border to the healthy bone after 7 days, later also lingually whereas no reaction was found laterally. The extent of the devitalised bone was 0.5-1.0 cm bigger than the cryosurgical part.

[Methemoglobinemia in the newborn infant--caused by prilocaine?-- A case report]. / Methämoglobinämie beim Neugeborenen--durch Prilocain verursacht?--Eine Falldarstellung.

The occurrence of postpartum cyanosis is primarily linked to pulmonary or cardiac causes. However, under differential diagnoses, methaemoglobinaemia should also be taken into consideration the causes of which may have toxical or hereditary origins. Diagnosis can be made by photospectrometical analysis of the methaemoglobin level in blood. Pathogenetical evidence is obtained by way of reduction method with methylen blue. This method will lead to consistently reliable results only, if the methaemoglobinaemia is of toxic nature. An example of toxically induced neonatal methaemoglobinaemia, which with great probability was caused by pudendus anaesthesia with Prilocain, is discussed in this case report.

[A new device technic for cryotherapy of glaucoma]. / Eine neue Gerätetechnik für die Kryotherapie des Glaukoms.

With nitrogen cryotherapy, the authors achieve lasting normalization of intraocular pressure in more than 90% of their primary glaucoma cases. To accomplish this, the therapeutic parameters (working temperature, diameter of the applicator tip, duration of application, number of application points, applicator contact pressure) must be strictly defined and coordinated. The applicator contact pressure has hitherto been neglected in published data, although it dictates the heat transfer resistance between the applicator and the surface of the bulb and is therefore a major factor influencing the therapeutic effect. A device was therefore developed which indicates the applicator contact pressure continuously during therapy, both optically and acoustically. It renders cryotherapy just as effective as microsurgical methods, with no side-effects.