Neuronal apoptosis by HIV-1 Vpr: contribution of proinflammatory molecular networks from infected target cells

Background: Human immunodeficiency virus type 1 (HIV-1) induces neuronal dysfunction through host cellular factors and viral proteins including viral protein R (Vpr) released from infected macrophages/microglia. Vpr is important for infection of terminally differentiated cells such as macrophages. The objective of this study was to assess the effect of Vpr in the context of infectious virus particles on neuronal death through proinflammatory cytokines released from macrophages.Methods: Monocyte-derived macrophages (MDM) were infected with either HIV-1 wild type (HIV-1wt), Vpr deleted mutant (HIV-1{increment}Vpr) or mock. Cell lysates and culture supernatants from MDMs were analyzed for the expression and release of proinflammatory cytokines by quantitative reverse transcription-PCR and enzyme-linked immunosorbent assay respectively. Mitogen-activated protein kinases (MAPK) were analyzed in activated MDMs by western blots. Further, the effect of Vpr on neuronal apoptosis was examined using primary neurons exposed to culture supernatants from HIV-1wt, HIV-1{increment}Vpr or mock-infected MDMs by Annexin-V staining, MTT and Caspase - Glo® 3/7 assays. The role of interleukin (IL)-1β, IL-8 and tumor necrosis factor (TNF)-α on neuronal apoptosis was also evaluated in the presence or absence of neutralizing antibodies against these cytokines.Results: HIV-1{increment}Vpr-infected MDMs exhibited reduced infection over time and specifically a significant downregulation of IL-1β, IL-8 and TNF-α at the transcriptional and/or protein levels compared to HIV-1wt-infected cultures. This downregulation was due to impaired activation of p38 and stress-activated protein kinase (SAPK)/c-Jun N-terminal kinase (JNK) in HIV-1{increment}Vpr-infected MDMs. The association of SAPK/JNK and p38 to IL-1β and IL-8 production was confirmed by blocking MAPKs that prevented the elevation of IL-1β and IL-8 in HIV-1wt more than in HIV-1{increment}Vpr-infected cultures. Supernatants from HIV-1{increment}Vpr-infected MDMs containing lower concentrations of IL-1β, IL-8 and TNF-α as well as viral proteins showed a reduced neurotoxicity compared to HIV-1wt-infected MDM supernatants. Reduction of neuronal death in the presence of anti-IL-1β and anti-IL-8 antibodies only in HIV-1wt-infected culture implies that the effect of Vpr on neuronal death is in part mediated through released proinflammatory factors.Conclusion: Collectively, these results demonstrate the ability of HIV-1{increment}Vpr to restrict neuronal apoptosis through dysregulation of multiple proinflammatory cytokines in the infected target cells either directly or indirectly by suppressing viral replication. © 2012 Guha et al.; licensee BioMed Central Ltd

Appearance of vascular endothelial growth factor (VEGF) in femoral head in the growing rat

In this study, we examined the appearance of vascular endothelial growth factor (VEGF) in the femoral head of the growing rat using an immunocytochemical technique. Our results showed VEGF-immunopositive cells existed in the inner region and peripheral region of the femoral head at each developmental stage. In the 19-day-old fetus, immunopositive mesenchymal cells were demonstrated in the peripheral region of the femoral head. At 1 to 10 days after birth, VEGF immunoreactivities were observed in the osteoblasts, osteoclasts, periosteum, perichondrium and cartilage matrix of the femur. At 15 days after birth, VEGF immunoreactive chondrocytes appeared in the apex area of the femoral head. In this stage, the femoral head is still constituted by chondrocytes and no apparent vascular formation has been observed. Thereafter, the immunopositive chondrocytes in the femoral head increased in number. The penetration of capillaries was recognized within the ligament of the femoral head at 60 days after birth. The results indicate that some chondrocytes in the femoral head produce VEGF before the beginning of ossification, and that VEGF may play an important role in the penetration of blood vessels into the femoral head from the ligament of the femoral head

Arsenic and boron in geothermal water and their removal

Selecting an As treatment technology for remediation of geothermal waters depends on several key factors. Among these, speciation of As, initial As concentration, regulatory requirements and target treatment levels must be considered. Due to variations in As speciation and large differences in the chemistry and physical properties of geothermal waters, no single technology will adequately meet the needs of every project. Furthermore, successful remediation often requires a combination of two or more treatment technologies. There are several inorganic arsenite species [inorganic As(III); e.g., H3AsO3, H2AsO3 Table 8.1 lists the As content of several geothermal waters in the world. Though large variation of As content is indicated in geothermal sites, extremely high As content is detected in some hot spring areas. This is linked to the mineralogical, chemical and physical characteristics of the soils, sediments and rocks in contact with these waters. In addition, the As is significantly leached from aquifers under the extremely high temperature and pressure conditions. Depending on oxidation-reduction (redox) conditions and biological activity, groundwater and geothermal water may contain As(V) and the more toxic As(III) forms (US EPA 2002). Considering that As contamination can originate from geological materials, the remediation of these materials is usually necessary to reduce As concentrations in associated geothermal waters. In some cases, however, geothermal water contamination is so severe that affordable and effective remediation is not possible. The physical and chemical characteristics of geothermal waters will affect the selection of reliable treatment technologies to work effectively under the high temperature condition. Alaerts and Khouri (2004) identified several factors that affect the costs and feasibility of treating As in geothermal water. The lowering of As drinking water standards (maximum contaminant level, MCL) from 50 to 10 µg L-1 in many countries has resulted in increasing demands for additional removal technologies when geothermal waters are used for drinking and cooking. © 2010 by Taylor & Francis Group, LLC

Appearance of vascular endothelial growth factor (VEGF) in femoral head in the growing rat

In this study, we examined the appearance of vascular endothelial growth factor (VEGF) in the femoral head of the growing rat using an immunocytochemical technique. Our results showed VEGF-immunopositive cells existed in the inner region and peripheral region of the femoral head at each developmental stage. In the 19-day-old fetus, immunopositive mesenchymal cells were demonstrated in the peripheral region of the femoral head. At 1 to 10 days after birth, VEGF immunoreactivities were observed in the osteoblasts, osteoclasts, periosteum, perichondrium and cartilage matrix of the femur. At 15 days after birth, VEGF immunoreactive chondrocytes appeared in the apex area of the femoral head. In this stage, the femoral head is still constituted by chondrocytes and no apparent vascular formation has been observed. Thereafter, the immunopositive chondrocytes in the femoral head increased in number. The penetration of capillaries was recognized within the ligament of the femoral head at 60 days after birth. The results indicate that some chondrocytes in the femoral head produce VEGF before the beginning of ossification, and that VEGF may play an important role in the penetration of blood vessels into the femoral head from the ligament of the femoral head

Selective recovery of lithium from seawater using a novel MnO2 type adsorbent III - benchmark evaluation

The granulation method of λ-MnO2 adsorbent employing chitin-based binder, which has efficient selectivity towards lithium ion, has been developed. The granules of ca. 1 – 2 mm with high resistance to the column operation for seawater (pH = 8.1) can be achieved. The laboratory scale column separation with the granulated adsorbent shows that lithium ions from seawater can be selectively recovered against the majority of co-existing cations. In addition, the elution of Mn from the adsorbent can be prevented. The benchmark column separation plant with seawater intake 200 L/h has been built and the whole process was verified and evaluated. The composition analysis of dried precipitated salts showed ca. 35 % efficiency of lithium recovery in the benchmark plant. In order to enhance the lithium recovery efficiency the following recovery steps are expected when routine techniques are applied

Coxsackievirus B3 genomes detected by polymerase chain reaction: evidence of latent persistency in the myocardium in experimental murine myocarditis

We have investigated the time course after infection in coxsackievirus B3 murine myocarditis to determine the extent (if any) of persistent or latent infection that might be responsible for recurrence. We employed a polymerase chain reaction (PCR) method that can detect an extremely small amount of genome by amplification techniques in four-week-old BALBIc mice (n=140) infected with coxsackievirus B3 by a single intraperitoneal injection of 1x10~? l aque-forming units (PFU)/mouse (Group 1) and 1x10 PFUJmouse (Group 2). Mice were sacrificed on days 3, 5, 7, 10, 14, 21 and 28, and their hearts were resected for RNA extraction. Single chain DNA was synthesized from 1 pg of RNA and the viral genome was amplified by PCR. The virus genome was strongly amplified in Group 1 from days 3 to 10, and in Group 2 from days 5 to 7, but afterwards both amplifications rapidly diminished. However, a positive signal, though very faint, persisted in both groups until day 28, by which time al1 histological evidence of myocarditis had disappeared in Group 2. Our results demonstrated that there was persistent or latent virus infection in the myocardium throughout the entire study period of 28 days. Such persistence might provide a pathomechanism for the exacerbation and recurrence of myocarditis