A defining characteristic of HIV-1 infection is the ability of the virus to persist within the host. Specifically, MHC-I downregulation by the HIV-1 accessory protein Nef is of critical importance in preventing infected cells from cytotoxic T-cell mediated killing. Nef downregulates MHC-I by modulating the host membrane trafficking machinery, resulting in the endocytosis and eventual sequestration of MHC-I within the cell. In the current report, we utilized the intracellular protein-protein interaction reporter system, bimolecular fluorescence complementation (BiFC), in combination with super-resolution microscopy, to track the Nef/MHC-I interaction and determine its subcellular localization in cells. We demonstrate that this interaction occurs upon Nef binding the MHC-I cytoplasmic tail early during endocytosis in a Rab5-positive endosome. Disruption of early endosome regulation inhibited Nef-dependent MHC-I downregulation, demonstrating that Nef hijacks the early endosome to sequester MHC-I within the
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Recent studies in the SIV-macaque model of HIV infection suggest that Nef-specific CD8+ T-cell responses may mediate highly effective immune control of viraemia. In HIV infection Nef recognition dominates in acute infection, but in large cohort studies of chronically infected subjects, breadth of T cell responses to Nef has not been correlated with significant viraemic control. Improved disease outcomes have instead been associated with targeting Gag and, in some cases, Pol. However analyses of the breadth of Nef-specific T cell responses have been confounded by the extreme immunogenicity and multiple epitope overlap within the central regions of Nef, making discrimination of distinct responses impossible via IFN-gamma ELISPOT assays. Thus an alternative approach to assess Nef as an immune target is needed. Here, we show in a cohort of |700 individuals with chronic C-clade infection that |50% of HLA-B-selected polymorphisms within Nef are associated with a predicted fitness cost to the virus, and that
Stimulation of secondary nef-specific CTLs by UbMNef and UbRNef constructs. PBMCs were isolated from an HIV-1 seropositive donor and stimulated in the presence
HIV-1 evades cytotoxic T cell responses through Nef-mediated downregulation of HLA class I molecules from the infected cell surface. Methods to quantify the impact of Nef on T cell recognition typically employ patient-derived T cell clones; however, these assays are limited by the cost and effort required to isolate and maintain primary cell lines. The variable activity of different T cell clones and the limited number of cells generated by re-stimulation can also hinder assay reproducibility and scalability.
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Wherry and his colleagues have also looked at the role of PD-1 in reducing T-cell responses. In a paper published recently in Nature Immunology2, Wherrys lab reported that inhibiting both PD-1 and another protein called LAG-3 boosted T-cell activity more than blocking either protein alone. But so far those experiments have only been carried out in mice. There are clearly several other pathways that operate in conjunction with PD-1, he says. Moving forward, wed like to see examination of these other pathways in SIV and HIV infection as well ...
2) write down all factor pairs of 4. (Note: since 5 is positive we only need to think about pairs that are either both positive or both negative. Remember a negative times a negative is a positve. As the chart on the right shows you -2*-2 is positive 4...so we do have to consider these two negative factors. This is probabily easier to understand if you watch our video lesson factoring trinonmials ...
HIV patients are at a greater risk of developing atherosclerosis than non-infected individuals, partly due to the impairment of the ATP-Binding Cassette A1 (ABCA1) cholesterol transporter by the HIV-1 viral protein Nef leading to accumulation of cholesterol inside the cell. While studying the possible mechanism of Nef-mediated disruption of cholesterol efflux, we found that ABCA1 interacts with Nef, but a direct interaction with Nef is dispensable for the inactivation of ABCA1. Using mass spectroscopy we identified calnexin as a protein that associates with both ABCA1 and Nef and provided evidence to show that in the presence of Nef, ABCA1-calnexin interaction is disrupted leading to ABCA1 retention in the ER, subsequent degradation and impairment of cholesterol efflux. However, the molecular interactions taking place remained unknown as Nef is not known to enter the ER lumen and the domain of calnexin involved in binding to substrate proteins is located within the ER lumen. We hypothesized that ...
Factor of infectivity and pathogenicity, required for optimal virus replication. Alters numerous pathways of T-lymphocytes function and down-regulates immunity surface molecules in order to evade host defense and increase viral infectivity. Alters the functionality of other immunity cells, like dendritic cells, monocytes/macrophages and NK cells.
Biochemical and cell-based studies indicate that Nef has multiple functions and that these functions are performed through multiple independent interactions with the host cell signal transduction and protein sorting machinery (29, 36, 45). This study indicates that a 239-Nef mutation which disrupts the interactions of Nef required for the downregulation of CD4 expression and for enhanced SIV replication in vitro also disrupts SIV replication in rhesus macaques. Not only were viral loads low early in macaques infected with the SIV containing the 239(EDR)-Nef variant, but there was also a strong selective pressure for revertants and second-site mutations which restored Nef function. The selection of these changes was associated with rises in viral loads, and virus recovered from these animals possessed virologic properties similar to those of wild-type SIV. While the EDR mutation disrupts other Nef functions, such as downregulation of CD28 cell surface expression (T. Swigut and J. Skowronski, ...
We studied the mechanisms underlying the down‐regulation of CD28 expression on the cell surface by HIV‐1 and SIV Nef proteins. Genetic and functional evidence from our studies indicates that Nef induces CD28 endocytosis via an AP‐2 clathrin adaptor‐dependent pathway, and that this is mediated by direct interactions between Nef, AP‐2 and CD28.. Nef probably makes direct contacts with the membrane‐proximal region in the CD28 cytoplasmic domain. This is suggested by the observation that induction of CD28 endocytosis by SIV and HIV‐1 Nef proteins requires partially overlapping but distinct sets of amino acid residues in the membrane‐proximal region of the CD28 cytoplasmic domain. The use of different target sites in the CD28 cytoplasmic domain by HIV‐1 and SIV Nef proteins argues against the possibility that they bind CD28 via a common bridging protein and implies that they contact the CD28 cytoplasmic domain directly.. Previous data indicate that Nef‐induced endocytosis of CD4 ...
In the original manuscript, the text in figure 1 is illegible. Furthermore, there is an unnecessary carriage return (page 1716, ~line 18) crystallographic ... methods. [...]
Jülich, 4 August 2017 - Around 37 million people around the globe are living with HIV. The illness associated with the infection, acquired immune deficiency syndrome (AIDS), can currently be treated with drugs - but is not yet curable. The human immunodeficiency virus (HIV) thus remains one of the greatest challenges for the public health system. The viral NEF (negative factor) protein is viewed as an important virulence factor contributing decisively to the course of the disease. Scientists at Jülichs Institute of Complex Systems and at the Düsseldorf Institut für Physikalische Biologie have now discovered that NEF requires a certain interaction partner in order to reach the plasma membrane of the host cell: the GABA(A) receptor-associated protein (GABARAP). When the researchers switch off this protein, NEF no longer reaches the inside of the plasma membrane. The article published in the journal Scientific Reports contributes to a better understanding of the functions of both the viral and human
HIV-1 Nef is normally an essential pathogenic aspect for HIV/AIDS pathogenesis. needed the membrane layer concentrating on function of Nef and was cell density-dependent. After that, we demonstrated that Nef transfer was cell-cell contact-dependent, as publicity to lifestyle supernatants or exosomes from HIV-infected Jurkat or Nef-expressing Jurkat and 293T led to small Nef recognition in […]. ...
HIV-1 isolate R3A Nef mutants G2, WL58, RR106, LL165, E160NNSLL165, and DD175 fail to induce release of IFN-alpha in pDCs, suggesting that the Nef function responsible for CD4 downregulation is crucial for pDCs stimulation by ...
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hello i was just wondering about some info about the rh factor in your blood. i am18 yrs old and 27 weeks pregnant and my doctor said i had a rh negative factor and only 15% of the pop. has it. So wha...
After a brief uptick at the opening bell, Australian shares fell Monday as a collection of negative factors held prices lower throughout the session.
Investigators have identified functional and structural nef gene defects that occur in HIV-infected paediatric patients with a non- or slow- progressing form of the disease, according to a report published in the November 20th issue of AIDS Research and Human Retroviruses. Dr. Rebeca Geffin, from the University of Miami School of Medicine, and colleagues analysed the structure and function of HIV-1 nef genes in seven children who were perinatally infected with the virus. Of this group, four patients were long-term nonprogressors, one was a slow progressor, and two were rapid progressors. Of the five slow progressors or nonprogressors, only one patient had large deletions of the nef gene, the authors note. The other four had nef genes of normal size but with a higher frequency of discrete changes than the genes of rapid progressors.. On functional analysis, the majority of Nef proteins derived from the study group were capable of binding…kinase p62, indicating that this function is well ...
Viruses encounter changing selective pressures during transmission between hosts, including host-specific immune responses and potentially varying functional demands on specific proteins. The human immunodeficiency virus type 1 Nef protein performs several functions potentially important for successful infection, including immune escape via down-regulation of class I major histocompatibility complex (MHC-I) and direct enhancement of viral infectivity and replication. Nef is also a major target of the host cytotoxic T-lymphocyte (CTL) response. To examine the impact of changing selective pressures on Nef functions following sexual transmission, we analyzed genetic and functional changes in nef clones from six transmission events. Phylogenetic analyses indicated that the diversity of nef was similar in both sources and acutely infected recipients, the patterns of selection across transmission were variable, and regions of Nef associated with distinct functions evolved similarly in sources and ...
Effects of HIV-1 Tat on expression of HLA class I molecules. Dendritic cells transduced with HIV Nef express normal levels of HLA-A and HLA-\b class I molecules
Peng, B.; Voltan, R.; Cristillo, A.D.; Alvord, W.Gregory.; Davis-Warren, A.; Zhou, Q.; Murthy, K.K.; Robert-Guroff, M., 2006: Replicating Ad-recombinants encoding non-myristoylated rather than wild-type HIV Nef elicit enhanced cellular immunity
Lentiviral Nef increases T cell signaling activity, but the molecular nature of the stimulus involved is incompletely described. We explored CD4 T cell lipid raft composition in the presence and absence of Nef. Here, the E2 ubiquitin-conjugating enzyme UbcH7, which acts in conjunction with c-Cbl, is absent from lipid rafts. This Nef-mediated exclusion is associated with failure of ubiquitination of activated Vav. In the presence of Nef, lipid raft Cdc42 is activated and forms a ternary complex between the c-Cbl-interacting protein p85Cool-1/betaPix and c-Cbl, displacing UbcH7 from rafts. Suppression of p85Cool-1/betaPix expression restores UbcH7 raft localization and Vav ubiquitination and diminishes Cdc42 activity. Moreover, p85Cool-1/betaPix knockdown attenuates HIV replication. Thresholds for activation of signaling involve the intricate balance of positive and negative regulators. Here we provide evidence for Nef disruption of a negative regulator of T cell signaling in promoting HIV replication.
Optimal HIV-1 infectivity requires the presence of both the viral factor Nef and the cellular protein cyclophilin A (CyPA) during virion assembly. These two proteins are integral components of HIV-1 particles. Both CyPA and Nef facilitate a step in the viral life cycle occurring between penetration …
University of Leeds Astbury Centre of Structural Molecular Biology and Division of Microbiology Research Fellowship in Structural Biology/Virology Closing date: 4 February 2000 The following positions are available within an expanding research group that is investigating the molecular basis of viral pathogenesis, particularly in relation to hepatitis C virus and HIV. Research Fellow: The structural and functional role of co- and post-translational modifications at the N-terminus of the HIV-1 Nef protein funded by the BBSRC for a period of 3 years from June 2000. Nef is a myristoylated phosphoprotein that is critical for viral pathogenesis. Whilst some structural information about the protein is available the influence of both myristoylation and phosphorylation on the structure of the native protein is unclear. You will therefore undertake structural studies on the native, myristoylated Nef protein expressed either using baculovirus or E.coli systems. This is a joint project between the ...
The PDB archive contains information about experimentally-determined structures of proteins, nucleic acids, and complex assemblies. As a member of the wwPDB, the RCSB PDB curates and annotates PDB data according to agreed upon standards. The RCSB PDB also provides a variety of tools and resources. Users can perform simple and advanced searches based on annotations relating to sequence, structure and function. These molecules are visualized, downloaded, and analyzed by users who range from students to specialized scientists.
Blocking that signal leaves people like those with AIDS - vulnerable to viruses, fungal infections and parasites, but especially micro-bacteria, a group of germs similar to tuberculosis that can cause severe lung damage. Researchers are calling this new disease an adult-onset immunodeficiency syndrome because it develops later in life and they dont know why or how ...
I remember a long while ago i told nef that they would make a great mentor and i still fully believe they would be a great purple mouse! Ive been in lots of shifts with nef and they are always great to see and talk to in a shift, they are friendly and kind when they interact with different people and also in discord its the same, i always see them talking with people and trying to help if someone has a problem or questions. in terms of knowledge i see them play a lot of sec and other jobs so i feel they definitely have lots of experience in many jobs and enough to be able to help lots of people! at the jobs they do they do very well too like sec, having nef on the team is always great and fun! maybe nef hos one day hehe ...
Within poor,women are affected more than men. In many societies, women bear the major responsibility of collection of fuel, fodder, water and other basic substance needs. AS these resources are depleted, the drudgery of women increases and sometimes they have to walk long distance which can cause serious health problems to them and negligence of home and children because of the increased hours of work ...
A class of antibiotic molecules called pleicomacrolides inhibit the Nef protein, which HIV uses to evade the bodys immune system.
We previously reported that the human immunodeficiency virus type 1 NL4-3 Nef is necessary and sufficient to induce a severe AIDS-like disease in transgenic (Tg) mice when the protein is expressed under the regulatory sequences of the human CD4 gene. We have now assayed additional Nef alleles (SF2, JR-CSF, YU10x, and NL4-3 [T71R] Nef alleles), including some from long-term nonprogressors (AD-93, 032an, and 039nm alleles) in the same Tg system and compared their pathogenicities. All these Nef alleles downregulated cell surface CD4 in human cells in vitro and also, with the exception of NefYU10x, in Tg CD4+ T cells. Depletion of double-positive and single-positive thymocytes occurred with all alleles but was less pronounced in NefYU10x Tg mice. A loss of peripheral CD4+ T cells was observed with all alleles but was minimal in NefYU10x Tg mice. In Nef032an and NefSF2 Tg mice, T-cell loss was severe despite lower levels of Tg expression, suggesting a higher virulence of these alleles. All Nef ...
The multifunctional Nef protein of HIV-1 is important for the progression to AIDS. One action of Nef is to down-regulate surface MHC I molecules, helping infected cells to evade immunity. We found that Nef also down-regulates the macrophage-expressed MHC 1b protein HFE, which regulates iron homeostasis and is mutated in the iron-overloading disorder hemochromatosis. In model cell lines, Nef reroutes HFE to a perinuclear structure that overlaps the trans-Golgi network, causing a 90% reduction of surface HFE. This activity requires a Src-kinase-binding proline-rich domain of Nef and a conserved tyrosine-based motif in the cytoplasmic tail of HFE. HIV-1 infection of ex vivo macrophages similarly down-regulates naturally expressed surface HFE in a Nef-dependent manner. The effect of Nef expression on cellular iron was explored; iron and ferritin accumulation were increased in HIV-1-infected ex vivo macrophages expressing wild-type HFE, but this effect was lost with Nef-deleted HIV-1 or when infecting
Human immunodeficiency computer virus (HIV) Nef is a membrane-associated protein decreasing surface expression of CD4 CD28 and major histocompatibility complex class I on infected cells. by the same mutations in Nef that impact CD4 down-regulation FK866 suggesting common molecular interactions. The ability to down-regulate the human CD8 β-chain was conserved in HIV-1 HIV-2 and simian immunodeficiency computer FK866 virus SIVmac239 Nef and required an intact AP-2 complex. The Nef-mediated internalization of receptors such as CD4 major histocompatibility complex class I CD28 and CD8αβ may contribute to the subversion of the host immune system and progression towards AIDS. The human immunodeficiency computer virus type 1 (HIV-1) Nef protein is usually a 27-kDa protein that is abundantly produced during the early phase of viral gene expression (28 54 Nef is usually posttranslationally altered by phosphorylation and due to irreversible attachment of myristic acid to its N terminus it is targeted ...
HIV-1 Nef critically contributes to AIDS in part by augmenting virus titers in infected individuals. Analyzing which of Nefs activities contribute to HIV pathogenesis has been hampered by the lack of a cell culture model in which Nef exerts pronounced effects on HIV replication. The human lymphoid aggregate culture (HLAC) from tonsil maintains the cell populations and cytokine milieu found in vivo, supports a productive infection without exogenous stimulation, and Nef contributes to efficient HIV-1 replication as well as CD4+ T cell depletion in this experimental ex vivo-model. To identify determinants in Nef that mediate these activities, we infected HLAC with a panel of isogenic HIV-1NL4-3 strains that encode for well-characterized mutants of HIV-1SF2 Nef. Determination of HIV-1 replication revealed that enhancement of the virus spread by Nef is governed by a complex set of protein interaction surfaces. In contrast, increased CD4+ T lymphocyte depletion depended on only two protein interaction
HIV-1 Nef critically contributes to AIDS in part by augmenting virus titers in infected individuals. Analyzing which of Nefs activities contribute to HIV pathogenesis has been hampered by the lack of a cell culture model in which Nef exerts pronounced effects on HIV replication. The human lymphoid aggregate culture (HLAC) from tonsil maintains the cell populations and cytokine milieu found in vivo, supports a productive infection without exogenous stimulation, and Nef contributes to efficient HIV-1 replication as well as CD4+ T cell depletion in this experimental ex vivo-model. To identify determinants in Nef that mediate these activities, we infected HLAC with a panel of isogenic HIV-1NL4-3 strains that encode for well-characterized mutants of HIV-1SF2 Nef. Determination of HIV-1 replication revealed that enhancement of the virus spread by Nef is governed by a complex set of protein interaction surfaces. In contrast, increased CD4+ T lymphocyte depletion depended on only two protein interaction
Defining the correlates of immune protection conferred by SIVΔnef, the most effective vaccine against SIV challenge, could enable the design of a protective vaccine against HIV infection. Here we provide a comprehensive assessment of immune responses that protect against SIV infection through detailed analyses of cellular and humoral immune responses in the blood and tissues of rhesus macaques vaccinated with SIVΔnef and then vaginally challenged with wild-type SIV. Despite the presence of robust cellular immune responses, animals at 5 weeks after vaccination displayed only transient viral suppression of challenge virus, whereas all macaques challenged at weeks 20 and 40 post-SIVΔnef vaccination were protected, as defined by either apparent sterile protection or significant suppression of viremia in infected animals. Multiple parameters of CD8 T cell function temporally correlated with maturation of protection, including polyfunctionality, phenotypic differentiation, and redistribution to gut ...
Participants expressing HIV antigens (gag, pol and nef) secrete antigen specific interferon-gamma (IFN-gamma). Levels of unfractionated gag, pol, and nef-specific IFN-gamma were to be measured using an Enzyme Linked Immunospot Assay (ELISPOT), which measures spot forming cells per 10^6 peripheral blood mononuclear cells (SFC per million PBMCs).. No immunogenicity analyses were performed because the results from a previous study, V520-023 (NCT00095576), which used the same vaccine as the one used in this study (NCT00849680) proved it was not efficacious. ...
Ho Chi Minh City health officials have denied a media report that people in the city have been infected with a new disease that US scientists say is similar to AIDS.
There are positive and negative factors that are discovered when selecting the desired site of a greenhouse. There are positive and negative factors that contribute to self-image as addressed in Objective 2 of this LK lesson. The skills to be learned is how to identify the controllable factors that lead to positive self image, just as a horticulturalists must identify what factors can be controlled to achieve proper site selection of a greenhouse. ...
Abcam provides specific protocols for Anti-HIV1 Nef antibody [3F2] (ab25866) : Western blot protocols, Immunoprecipitation protocols
Listen to DaBoyDame, Blac Youngsta & Mozzy featuring Nef The Pharaoh Look At Me Now. DaBoyDame linked up with two of the biggest street rappers, Blac Young...
The purpose of the study is to justify selection of a certain EOR technology based on the results of its practical application. The results of the study are needed to plan economically effective operation of TATNEFT Company in the short term. The paper describes processes taking place in a reservoir when different chemical agents have been injected. Positive and negative factors affecting the efficiency of technologies are described; results of different technologies in the same initial conditions are evaluated. The authors analyze the performance of different EOR technologies in the same area and in the same well.. ...
Hi everybody? Have any of you managed to get Finder to display the thumbnails it generates in Icon mode (CMND+1) to respect their modified...
台灣M·A·C的愛滋關懷浪潮中從不缺席。截至目前為止,台灣M.A.C已募得2千萬2百萬元(共3萬6千支VIVA GLAM薇拉葛蘭唇膏,累積高度相當於6個台北101 ),藉由「薇拉葛蘭VIVA GLAM系列」100%捐贈的力量,M·A·C愛滋公益活動成為台灣最具代表性的公益盛事,實際援助台灣愛滋團體,如關愛之家、露德協會…等,讓HIV/AIDS朋友們擁有最直接的資源與協助。台灣M·A·C特別邀請華人音樂時尚天后 - 蕭亞軒 Elva ...
A highly conserved threonine near the C terminus of gp120 of human immunodeficiency virus (HIV) and simian immunodeficiency virus (SIV) was investigated for its contributions to envelope protein function and virion infectivity. When this highly conserved Thr residue was substituted with anything other than serine (the other amino acid that can accept O-glycosylation), the resulting virus was noninfectious. We found that this Thr was critical for the association of gp120 with the virion and that amino acid substitution increased the amount of dissociated gp120 in the cell culture supernatant. When HIV virions were generated in cells overexpressing polypeptide N-acetylgalactosaminyltransferase 1 (GalNAcT1), viral infectivity was increased 2.5-fold compared to that of virus produced in wild-type HEK293T cells; infectivity was increased 8-fold when the Thr499Ser mutant was used. These infectivity enhancements were not observed when GalNAcT3 was used. Using HEK293T knockout cell lines totally devoid ...
An additional series of mutational analyses revealed a fine specificity for the lipopeptide recognition by the T cells. The 5-mer Nef peptide conjugated with a shorter (C10) saturated fatty acid (C10-GGAIS) showed reduced T cell stimulation activity compared with C14nef5, and no T cell response was detected for C6-GGAIS (Fig. 1F), further confirming that the peptide modification with a fatty acid of the C14 chain length (myristic acid) was essential for activation of the 2N5.1 cells. The N-terminal amino acid sequence (GGAIS) of the Nef protein matches with a typical N-myristoylation motif, Gly-X-X-X-(Ser/Thr), in which X is any amino acid (3). Whereas the serine-to-threonine substitution (C14-GGAIT) did not affect the antigenic activity, alanine substitution for either the second glycine residue (C14-GAAIS) or the isoleucine residue (C14-GGAAS) located between the conserved flanking amino acid residues totally abrogated the activity (Fig. 1F). Furthermore, addition of an amide linkage to the ...
HIV-1 Nef clones isolated from a rare subset of HIV-1-infected elite controllers (EC), with the ability to suppress viral load to undetectable levels in the absence of antiretroviral therapy, are unable to fully downregulate CD4 from the plasma membrane of CD4(+) T cells. Residual CD4 left at the plasma membrane allows Env-CD4 interaction, which leads to increased exposure of Env CD4-induced epitopes and increases susceptibility of infected cells to antibody-dependent cellular cytotoxicity (ADCC). ADCC is mediated largely by natural killer (NK) cells, which control their activation status through the cumulative signals received through activating and inhibitory receptors. Recently, the activating NKG2D receptor was demonstrated to positively influence ADCC responses. Since HIV-1 Nef has been reported to reduce the expression of NKG2D ligands, we evaluated the relative abilities of Nef from EC and progressors to downmodulate NKG2D ligands. Furthermore, we assessed the impact of EC and progressor ...
by Tim Horn, Senior Writer & Editor, AIDSmeds.com. HIVs ability to damage the human immune system might amount to an accident of evolution, notably the loss of the protective function of a viral protein called Nef. Like HIV in humans, related strains of simian immunodeficiency virus (SIV) are rampant among many species of monkeys. Unlike HIV in humans, many primates infected with SIV dont experience immune suppression or suffer the symptoms associated with AIDS. The evidence, published by an international team of researchers in the June 2006 issue of Cell, is the first to offer an explanation for this striking difference. The group found that a viral protein known to help the virus evade the immune system, thereby allowing the SIVs that infect monkeys to persist and multiply with high efficiency, also has a protective role in the host immune system. The SIV Nef protein ratchets down the activation of T-cells following infection in primates, thereby limiting the harmful effects that can be ...
No one ever really discussed the danger of mouthwashes and how it can be a negative factor to a persons health - heres more info about it.
Hello Everyone! I am considering building up a short carbine as a walking-around gun, based upon a new, steel-receiver H&R/NEF Handi-Rifle. The barrel...