Brca1 is expressed in human microglia and is dysregulated in human and animal model of ALS

Background: There is growing evidence that microglia are key players in the pathological process of amyotrophic lateral sclerosis (ALS). It is suggested that microglia have a dual role in motoneurone degeneration through the release of both neuroprotective and neurotoxic factors. Results: To identify candidate genes that may be involved in ALS pathology we have analysed at early symptomatic age (P90), the molecular signature of microglia from the lumbar region of the spinal cord of hSOD1(G93A) mice, the most widely used animal model of ALS. We first identified unique hSOD1(G93A) microglia transcriptomic profile that, in addition to more classical processes such as chemotaxis and immune response, pointed toward the potential involvement of the tumour suppressor gene breast cancer susceptibility gene 1 (Brca1). Secondly, comparison with our previous data on hSOD1(G93A) motoneurone gene profile substantiated the putative contribution of Brca1 in ALS. Finally, we established that Brca1 protein is specifically expressed in human spinal microglia and is up-regulated in ALS patients. Conclusions: Overall, our data provide new insights into the pathogenic concept of a non-cell-autonomous disease and the involvement of microglia in ALS. Importantly, the identification of Brca1 as a novel microglial marker and as possible contributor in both human and animal model of ALS may represent a valid therapeutic target. Moreover, our data points toward novel research strategies such as investigating the role of oncogenic proteins in neurodegenerative diseases.

The coastal environment and human health: microbial indicators, pathogens, sentinels and reservoirs

Innovative research relating oceans and human health is advancing our understanding of disease-causing organisms in coastal ecosystems. Novel techniques are elucidating the loading, transport and fate of pathogens in coastal ecosystems, and identifying sources of contamination. This research is facilitating improved risk assessments for seafood consumers and those who use the oceans for recreation. A number of challenges still remain and define future directions of research and public policy. Sample processing and molecular detection techniques need to be advanced to allow rapid and specific identification of microbes of public health concern from complex environmental samples. Water quality standards need to be updated to more accurately reflect health risks and to provide managers with improved tools for decision-making. Greater discrimination of virulent versus harmless microbes is needed to identify environmental reservoirs of pathogens and factors leading to human infections. Investigations must include examination of microbial community dynamics that may be important from a human health perspective. Further research is needed to evaluate the ecology of non-enteric water-transmitted diseases. Sentinels should also be established and monitored, providing early warning of dangers to ecosystem health. Taken together, this effort will provide more reliable information about public health risks associated with beaches and seafood consumption, and how human activities can affect their exposure to disease-causing organisms from the oceans.

Phantom mutation hotspots in human mitochondrial DNA

Phantom mutations are systematic artifacts generated in the course of the sequencing process. Contra common belief these artificial mutations are nearly ubiquitous in sequencing results, albeit at frequencies that may vary dramatically. The amount of artifacts depends not only on the sort of automated sequencer and sequencing chemistry employed, but also on other lab-specific factors. An experimental study executed on four samples under various combinations of sequencing conditions revealed a number of phantom mutations occurring at the same sites of mitochondrial DNA (mtDNA) repeatedly. To confirm these and identify further hotspots for artifacts, > 5000 mtDNA electropherograms were screened for artificial patterns. Further, > 30000 published hypervariable segment 1 sequences were compared at potential hotspots for phantom mutations, especially for variation at positions 16085 and 16197. Resequencing of several samples confirmed the artificial nature of these and other polymorphisms in the original publications. Single-strand sequencing, as typically executed in medical and anthropological studies, is thus highly vulnerable to this kind of artifacts. In particular, phantom mutation hotspots could easily lead to misidentification of somatic mutations and to misinterpretations in all kinds of clinical mtDNA studies.

Survey of plankton blooms and red tides and their interaction with environmental factors through monitoring satellite images

One of the most important marine ecological phenomena is red tide which is created by increasing of phytoplankton population, influenced by different factors such as climate condition changes, utrification hydrological factors and can leave sever and undesired ecological and economical effects behind itself in the case of durability. Coast line of Hormozgan is about 900km from east to west, within the range of geographical coordinates of 56 16 23.8, 26 58 8.8 to 54 34 5.33 and 26 34 32 eastern longitude and northern latitude, seven sampling stations were considered and sampled for a period of one year from October 2008 to October 2009. after the analysis of Satellite images, monthly, during the best time. In several stages, samplings were performed. In each station, three samples were collected for identification and determination of Bloom- creating species abundance. Cochlodinium polykrikoides was the species responsible for the discoloration which occurred at October 2008 in Hormozgan marine water. Environmental parameters such as sea surface temperature, pH, salinity, Dissolved Oxygen concentration, Total Dissolved Solids (T.D.S.), conductivity, nitrate, nitrite and phosphate and also chlorophyll a were measured and calculated. Kruscal Wallis test was used to compare the densities between different months, seasons and the studied stations. Mann-whitney test from Nonparametric Tests was used for couple comparison. Pearson correlation coefficient was used to determine the relationship between physical and chemical data set and the abundance of Cochlodinium polykrikoides. Multivariate Regression and analysis of variance (ANOVA) also were used to obtain the models and equations of red tide occurrence relationship, environmental parameters and nutrient data. The highest density was 26 million cells per liter in Qeshm station. A meaningful difference was observed between sampling months and seasons but there was no between sampling stations which indicates that in favorable conditions, the occurrence of this phenomenon by the studied species is probable. Regarding to β coefficients of nitrate, temperature, phosphate, Total Dissolvable Solutions (T.D.S) and pH these parameters are effective on the abundance of this species and red tide occurrence. Increase in these factors can represent the effects and outcomes of human activities and increase in marine pollution.

The effect of spirulina (fresh and dry) on some biological factors in Penaeus semisulcatus larvae

Spirulina is a filamentous cyanobacteria with many applications in food and drug industries, as a food in human, aquaculture, vet and poultry industries… . Semi and mass culture of Spirulina carries out in different countries. This study was carried out in five phases in order to produce this microalga in Iran. The first phase, Spirulina pure stock was imported from Indonesia. After identification of species, it was cultured in laboratory until we took 20 liters of biomass. The semi-mass culture was carried out in green house. Cell concentration and size of Spirulina were recorded during culture daily and their growth rates were calculated. After two weeks, when the size of Spirulina was suitable, biomass of Spirulina was harvested then accumulated Spirulina weighted and dried in 24 hours in laboratory. In order to microbiological study, the samples of Spirulina (dry and fresh) were cultured on blood agar medium and coliforms were counted. The chemical composition of produced Spirulina was measured by standard methods. Fatty acid and amino acid profiles were acquired by GC and HPLC instruments, respectively. The amount of chlorophyll in Spirulina was determined by spectroscopy method. Also astaxanthin pigment as an important carotenoid was measured by HPLC in Spirulina and Penaeus semisulcatus larvae fed on Spirulina. At final phase of this project, larva fed on produced Spirulina (biomass and dry powder) was compared to Z plus, microencapsulated Spirulina (M.C.F) and Chaetoceros algae as control. This experiment was carried from zoa to early post larvae stage then survival and growth rate of larvae were recorded. The growth rate of larvae was evaluated with ANOVA test and survival rate of treatments was assessed by Log Rank (Mantel –Cox) test. Also during larvae stage, two parameters of water such as nitrate and nitrite were measured in zoa, mysis and post larvae stages. The results of this study were shown that colifom counts were 1.85×106 and 92.3×105 coliform per ml in fresh and dry spirulina, respectively. Protein percent of dry spirulina was 50.93 % (dry weight) and the amount of astaxanthin in spirulina and larvae fed on spirulina were 0.21 and 0.01 mg/kg, respectively. The most survival rate of larvae were observed in zoa III (88.8%) with Z plus supplement treatment, in mysis III (76.5%) combination of Z plus and dry spirulina in comparative between treatments. Larvae growth (4.5mm) of control in early post larvae was the best.

Impacts of socio-economic factors on sediment yield in the Upper Yangtze River

In recent years, the role of human activities in changing sediment yield has become more apparent for the construction of hydraulic engineering and water conservation projections in the Upper Yangtze River, but it has not been evaluated at the macro scale. Taking Sichuan Province and Chongqing City as an example, this paper studies the relationship between socio-economic factors and sediment yield in the Upper Yangtze River based on section data in 1989 and 2007. The results show that sediment yield is significantly correlated with population density and cultivated area, in which the former appears to be more closely related to sediment yield. Moreover, in the relation of sediment yield vs. population density, a critical value of population density exists, below which the sediment yield increases with the increase of population density and over which the sediment yield increases with the decrease of population density. The phenomenon essentially reflects the influence of natural factors, such as topography, precipitation and soil property, and some human activities on sediment yield. The region with a higher population density than critical value is located in the east of the study area and is characterized by plains, hills and low mountains, whereas the opposite is located in the west and characterized by middle and high mountains. In the eastern region, more people live on the lands with a low slope where regional soil erosion is slight; therefore, sediment yield is negatively related with population density. In contrast, in the western region, the population tends to aggregate in the areas with abundant soil and water resources which usually lead to a higher intensity of natural erosion, and in turn, high-intensity agricultural practices in these areas may further strengthen local soil erosion. It is also found that population tends to move from the areas with bad environment and high sediment yield to the areas with more comfortable environment and less sediment yield. The natural factors have greater influence on sediment yield of western region than that of eastern region. Generally, the natural factors play a dominant role on sediment yield in the Upper Yangtze River.

Affinity and Specificity of Levamlodipine-Human Serum Albumin Interactions: Insights into Its Carrier Function

The affinity and specificity of drugs with human serum albumin (HSA) are crucial factors influencing the bioactivity of drugs. To gain insight into the carrier function of HSA, the binding of levamlodipine with HSA has been investigated as a model system by a combined experimental and theoretical/computational approach. The fluorescence properties of HSA and the binding parameters of levamlodipine indicate that the binding is characterized by one binding site with static quenching mechanism, which is related to the energy transfer. As indicated by the thermodynamic analysis, hydrophobic interaction is the predominant force in levamiodipine-HSA complex, which is in agreement with the computational results. And the hydrogen bonds can be confirmed by computational approach between levamlodipine and HSA. Compared to predicted binding energies and binding energy spectra at seven sites on HSA, levamlodipine binding HSA at site I has a high affinity regime and the highest specificity characterized by the largest intrinsic specificity ratio (ISR). The binding characteristics at site I guarantee that drugs can be carried and released from HSA to carry out their specific bioactivity.

Direct electrochemical detection of glucose in human plasma on capillary electrophoresis microchips

We developed an electrochemical detector on a hybrid chip for the determination of glucose in human plasma. The microchip system described in this paper consists of a poly(dimethylsiloxane) (PDMS) layer containing separation and injection channels and an electrode plate. The copper microelectrode is fabricated by selective electroless deposition. The fabrication of the decoupler is performed by platinum electrochemical deposition on the metal film formed by electroless deposition. Factors influencing the performance, including detection potential, separation field strength, and buffer concentration, were studied. The electrodes exhibited good stability and durability in the analytical procedures. Under optimized detection conditions, glucose responded linearly from 10 muM to 1 mM. Finally, glucose in human plasma from three healthy individuals and two diabetics was successfully determined, giving a good prospect for a new clinical diagnostic instrument.

Factors that effect antioxidant activity of C-phycocyanins from Spirulina platensis

Currently, antioxidants are added in the human diet to prevent free radical-induced cell damage, and there has been an explosive interest in the use of antioxidant nutritional supplements. The effects of different factors on the antioxidant activity of phycocyanins (PCs) were studied. The results showed that PCs generated hydroxyl radicals in the light, while scavenging them in the dark. When PCs were denatured by sodium dodecyl sulfate, urea and in alkaline condition, their ability to generate hydroxyl radicals disappeared and that of scavenging them greatly increased. This showed that the phycobilin moiety is the main part of PC involved in scavenging hydroxyl radicals. Trypsin hydrolysis of PCs showed that the apoprotein portion of the molecule also made a significant contribution to the antioxidant activity.

Community structure of macrobenthos in coastal water off Rushan, southern Shandong Peninsula, and the relationships with environmental factors

To understand the present actuality of the marine ecosystem in the southern coastal water region of the Shandong Peninsula and the impact of the global change and the human activities to the marine ecosystem of the region, the macrobenthic community structure was researched based on data from 26 sampling stations carried out on four seasonal cruises from December 2006 to November 2007. The data was analyzed using PRIMER 6.0 and SPSS 15.0 software packages. The results showed that 236 macrobenthic species in total were collected from the research region by the field works. Most of the species belong to Polychaeta (76 species), Mollusca (75) and Crustacea (60). Of which, 33 species were common species by the four cruises. The dominant species were different among the four seasons, however, the polychaete species Nephtys oligobranchia and Sternaspis scutata were always dominant in the four seasons. The abundances and biomasses of the macrobenthos from the research region were variable in tire four seasons. The results of CLUSTER and MDS analysis showed that the similarities of macrobenthic structures among the stations were low, most of the similarities were at about 40% of similarity values, only that of two stations were up to 60%. In accordance with the similarity values of the macrobenthic structures, the 26 stations were clustered as six groups at arbitrary similarity level of 30%. The ABC curve indicated that the marcofauna communities in the research region had riot been disturbed distinctly. The results of BIOENV and BVSTEP (Spearman) analysis implied that the concentrations of organic matter in bottom water and heavy metal copper in sediment, water depth and temperature of bottom were the most significant environmental factors to affect the macrobentic community.

Expression of C-reactive protein in human lung epithelial cells and up-regulation by cytokines and carbon particles.

C-reactive protein (CRP) is the prototypic human acute-phase protein and is found at increased levels in the blood during episodes of inflammation. CRP was generally thought to be produced only by hepatocytes; however, several studies have shown extrahepatic synthesis of CRP. A previous study showed that PM10 and ultrafine carbon black (ufCB) were able to induce CRP expression in A549 cells. This study aims to examine the factors that lead to the production of CRP in A549 cells. A549 human lung epithelial cells were treated with cytokines (interleukin 6, tumor necrosis factor , interferon , or interleukin 1) or carbon particles (CB and ufCB) for 18 h. It was found that CRP could be expressed within the cells and that CRP was secreted from the cells particularly with tumor necrosis factor , CB and ufCB treatments. It was also found that this expression of CRP with CB and ufCB treatments was dependent on nuclear factor kappa B (NFB). The expression of CRP in A549 cells may indicate an important role for CRP expression and secretion from lung epithelial cells in response to inflammatory stimuli.

Human papillomavirus detection in women with and without human immunodeficiency virus infection in Colombia

Background: HIV infection leads to a decreasing immune response, thereby facilitating the appearance of other infections, one of the most important ones being HPV. However, studies are needed for determining associations between immunodeficiency caused by HIV and/or the presence of HPV during the course of cervical lesions and their degree of malignancy. This study describes the cytological findings revealed by the Papanicolaou test, laboratory characteristics and HPV molecular profile in women with and without HIV infection. Methods: A total of 216 HIV-positive and 1,159 HIV-negative women were invited to participate in the study; PCR was used for the molecular detection of HPV in cervical samples. Statistical analysis (such as percentages, Chi-square test and Fisher's exact test when applicable) determined human papillomavirus (HPV) infection frequency (single and multiple) and the distribution of six types of high-risk-HPV in women with and without HIV infection. Likewise, a logistic regression model was run to evaluate the relationship between HIV-HPV infection and different risk factors. Results: An association was found between the frequency of HPV infection and infection involving 2 or more HPV types (also known as multiple HPV infection) in HIV-positive women (69.0% and 54.2%, respectively); such frequency was greater than that found in HIV-negative women (44.3% and 22.7%, respectively). Statistically significant differences were observed between both groups (p = 0.001) regarding HPV presence (both in infection and multiple HPV infection). HPV-16 was the most prevalent type in the population being studied (p = 0.001); other viral types had variable distribution in both groups (HIV-positive and HIV-negative). HPV detection was associated with <500 cell/mm(3) CD4-count (p = 0.004) and higher HIV-viral-load (p = 0.001). HPV-DNA detection, <200 cell/mm(3) CD4-count (p = 0.001), and higher HIV-viral-load (p = 0.001) were associated with abnormal cytological findings. Conclusions: The HIV-1 positive population in this study had high multiple HPV infection prevalence. The results for this population group also suggested a greater association between HPV-DNA presence and cytological findings. HPV detection, together with low CD4 count, could represent useful tools for identifying HIV-positive women at risk of developing cervical lesions.

Human interfaces to structured documents

This research investigates some of the reasons for the reported difficulties experienced by writers when using editing software designed for structured documents. The overall objective was to determine if there are aspects of the software interfaces which militate against optimal document construction by writers who are not computer experts, and to suggest possible remedies. Studies were undertaken to explore the nature and extent of the difficulties, and to identify which components of the software interfaces are involved. A model of a revised user interface was tested, and some possible adaptations to the interface are proposed which may help overcome the difficulties. The methodology comprised: 1. identification and description of the nature of a ‘structured document’ and what distinguishes it from other types of document used on computers; 2. isolation of the requirements of users of such documents, and the construction a set of personas which describe them; 3. evaluation of other work on the interaction between humans and computers, specifically in software for creating and editing structured documents; 4. estimation of the levels of adoption of the available software for editing structured documents and the reactions of existing users to it, with specific reference to difficulties encountered in using it; 5. examination of the software and identification of any mismatches between the expectations of users and the facilities provided by the software; 6. assessment of any physical or psychological factors in the reported difficulties experienced, and to determine what (if any) changes to the software might affect these. The conclusions are that seven of the twelve modifications tested could contribute to an improvement in usability, effectiveness, and efficiency when writing structured text (new document selection; adding new sections and new lists; identifying key information typographically; the creation of cross-references and bibliographic references; and the inclusion of parts of other documents). The remaining five were seen as more applicable to editing existing material than authoring new text (adding new elements; splitting and joining elements [before and after]; and moving block text).

Identification of major factors influencing ELISpot-based monitoring of cellular responses to antigens from Mycobacterium tuberculosis.

A number of different interferon-gamma ELISpot protocols are in use in laboratories studying antigen-specific immune responses. It is therefore unclear how results from different assays compare, and what factors most significantly influence assay outcome. One such difference is that some laboratories use a short in vitro stimulation period of cells before they are transferred to the ELISpot plate; this is commonly done in the case of frozen cells, in order to enhance assay sensitivity. Other differences that may be significant include antibody coating of plates, the use of media with or without serum, the serum source and the number of cells added to the wells. The aim of this paper was to identify which components of the different ELISpot protocols influenced assay sensitivity and inter-laboratory variation. Four laboratories provided protocols for quantifying numbers of interferon-gamma spot forming cells in human peripheral blood mononuclear cells stimulated with Mycobacterium tuberculosis derived antigens. The differences in the protocols were compared directly. We found that several sources of variation in assay protocols can be eliminated, for example by avoiding serum supplementation and using AIM-V serum free medium. In addition, the number of cells added to ELISpot wells should also be standardised. Importantly, delays in peripheral blood mononuclear cell processing before stimulation had a marked effect on the number of detectable spot forming cells; processing delay thus should be minimised as well as standardised. Finally, a pre-stimulation culture period improved the sensitivity of the assay, however this effect may be both antigen and donor dependent. In conclusion, small differences in ELISpot protocols in routine use can affect the results obtained and care should be given to conditions selected for use in a given study. A pre-stimulation step may improve the sensitivity of the assay, particularly when cells have been previously frozen.

Differential inhibition of human immunodeficiency virus type 1 in peripheral blood mononuclear cells and TZM-bl cells by endotoxin-mediated chemokine and gamma interferon production.

Bacterial lipopolysaccharide (endotoxin) is a frequent contaminant of biological specimens and is also known to be a potent inducer of beta-chemokines and other soluble factors that inhibit HIV-1 infection in vitro. Though lipopolysaccharide (LPS) has been shown to stimulate the production of soluble HIV-1 inhibitors in cultures of monocyte-derived macrophages, the ability of LPS to induce similar inhibitors in other cell types is poorly characterized. Here we show that LPS exhibits potent anti-HIV activity in phytohemagglutinin-stimulated peripheral blood mononuclear cells (PBMCs) but has no detectable anti-HIV-1 activity in TZM-bl cells. The anti-HIV-1 activity of LPS in PBMCs was strongly associated with the production of beta-chemokines from CD14-positive monocytes. Culture supernatants from LPS-stimulated PBMCs exhibited potent anti-HIV-1 activity when added to TZM-bl cells but, in this case, the antiviral activity appeared to be related to IFN-gamma rather than to beta-chemokines. These observations indicate that LPS stimulates PBMCs to produce a complex array of soluble HIV-1 inhibitors, including beta-chemokines and IFN-gamma, that differentially inhibit HIV-1 depending on the target cell type. The results also highlight the need to use endotoxin-free specimens to avoid artifacts when assessing HIV-1-specific neutralizing antibodies in PBMC-based assays.