Genetic Analysis of Cytokine Promoters in Nonhuman Primates: Implications for Th1/Th2 Profile Characteristics and SIV Disease Pathogenesis

The shift from a predominant synthesis of prototype Th1 cytokines to Th2 or Th0 type of cytokines by antigen activated PBMC's from HIV infected humans and SIV infected disease susceptible rhesus macaques (RM) has been shown to be associated with disease progression. Paradoxically, antigen activated PBMC's from sooty mangabeys (SM), which are naturally infected with SIV and are disease resistant despite high viral loads, maintain a predominant Th2 cytokine profile. It has been reasoned that the resistance to perturbations of cytokine synthesis by slow and/or nonprogressor HIV infected patients and SIV infected disease susceptible RM is secondary to inherited polymorphisms within the promoter regions for cytokines. Similar promoter polymorphisms could also contribute to the cytokine profile of PBMC's from SM. To address this issue promoter regions for the major Th1/Th2 cytokines from RM and SM were cloned and sequenced. Sequence analysis of promoter fragments of IL-4, IL-10, IL-12 p40, IFN-gamma and TNF-alpha from the two monkey species showed varying degree of homology ranging from high degree of homology detected for IFN-gamma promoter (>99%) to relatively high degree of polymorphism detected for TNF-alpha promoter (94% homology). In addition, several variable regions within the promoters of IL-12 p40, IL-10 and TNF-alpha in the two species contain polymorphisms in sequences that constitute binding sites of known transcription factors (TF). Such differences are likely to differentially bind TF and thus either qualitatively and/or quantitatively affect the regulation of cytokine synthesis in these two species and potentially contribute to disease progression and/or resistance

Distinct host cell proteins incorporated by SIV replicating in CD4+ T Cells from natural disease resistant versus non-natural disease susceptible hosts

<p>Abstract</p> <p>Background</p> <p>Enveloped viruses including the simian immunodeficiency virus (SIV) replicating within host cells acquire host proteins upon egress from the host cells. A number of studies have catalogued such host proteins, and a few have documented the potential positive and negative biological functions of such host proteins. The studies conducted herein utilized proteomic analysis to identify differences in the spectrum of host proteins acquired by a single source of SIV replicating within CD4⁺T cells from disease resistant sooty mangabeys and disease susceptible rhesus macaques.</p> <p>Results</p> <p>While a total of 202 host derived proteins were present in viral preparations from CD4⁺T cells from both species, there were 4 host-derived proteins that consistently and uniquely associated with SIV replicating within CD4⁺T cells from rhesus macaques but not sooty mangabeys; and, similarly, 28 host-derived proteins that uniquely associated with SIV replicating within CD4⁺T cells from sooty mangabeys, but not rhesus macaques. Of interest was the finding that of the 4 proteins uniquely present in SIV preparations from rhesus macaques was a 26 S protease subunit 7 (MSS1) that was shown to enhance HIV-1 'tat" mediated transactivation. Among the 28 proteins found in SIV preparations from sooty mangabeys included several molecules associated with immune function such as CD2, CD3ε, TLR4, TLR9 and TNFR and a bioactive form of IL-13.</p> <p>Conclusions</p> <p>The finding of 4 host proteins that are uniquely associated with SIV replicating within CD4⁺T cells from disease susceptible rhesus macaques and 28 host proteins that are uniquely associated with SIV replicating within CD4⁺T cells from disease resistant sooty mangabeys provide the foundation for determining the potential role of each of these unique host-derived proteins in contributing to the polarized clinical outcome in these 2 species of nonhuman primates.</p

Reactivation of Human Acetylcholinesterase and Butyrylcholinesterase Inhibited by Leptophos-Oxon with Different Oxime Reactivators in Vitro

We have evaluated in vitro the potency of 23 oximes to reactivate human erythrocyte acetylcholinesterase (AChE) and plasma butyrylcholinesterase (BChE) inhibited by racemic leptophos-oxon (O-[4-bromo-2,5-dichlorophenyl]-O-methyl phenyl-phosphonate), a toxic metabolite of the pesticide leptophos. Compounds were assayed in concentrations of 10 and 100 μM. In case of leptophos-oxon inhibited AChE, the best reactivation potency was achieved with methoxime, trimedoxime, obidoxime and oxime K027. The most potent reactivators of inhibited BChE were K033, obidoxime, K117, bis-3-PA, K075, K074 and K127. The reactivation efficacy of tested oximes was lower in case of leptophos-oxon inhibited BChE

The Development of Mouse APECED Models Provides New Insight into the Role of AIRE in Immune Regulation

Autoimmune polyendocrinopathy candidiasis ectodermal dystrophy is a rare recessive autoimmune disorder caused by a defect in a single gene called AIRE (autoimmune regulator). Characteristics of this disease include a variable combination of autoimmune endocrine tissue destruction, mucocutaneous candidiasis and ectodermal dystrophies. The development of Aire-knockout mice has provided an invaluable model for the study of this disease. The aim of this review is to briefly highlight the strides made in APECED research using these transgenic murine models, with a focus on known roles of Aire in autoimmunity. The findings thus far are compelling and prompt additional areas of study which are discussed

Simple Method for Selective Amplification of cDNA from a Defined Promoter

A simplified technique for the detection of transcripts from a defined promoter is described. After reverse transcription, a PCR target sequence is selectively added to the 3′ end of cDNA strands by DNA polymerase extension directed by an oligonucleotide template. Those cDNA molecules that do not have ends within a few nucleotides of the promoter start site are not extended and thus are excluded from subsequent amplification. Even when amplified products are visualized by ethidium bromide staining of agarose gels, this method requires only 1% of the RNA usually needed for detection of mRNA by standard RNase protection utilizing radiolabeled probes. In contrast to direct detection of cDNA by PCR, this procedure restricts amplification to a narrow subset of transcripts even when other overlapping colinear transcripts are present. We call this detection procedure specific amplification of cDNA ends (SPACE)

Czech Honeydew Honeys—A Potential Source of Local Medical Honey with Strong Antimicrobial Activity

An increasing resistance of microbes to antibiotics, the emergence of multidrug-resistant and extremely resistant strains, and the long time needed to develop new antibiotics are driving the search for additional sources of antibacterial agents. The aim of the study was to compare the efficacy of Czech honeys with already available pharmaceutical agents containing medicinal honey, and to perform basic biochemical analysis of Czech samples, including detection of undesirable chemical substances. The results showed strong antibacterial activity of Czech honeydew honeys compared to the control group, especially against G+ pathogens, with an average MIC of 9.44% compared to 17.54%, and comparable activity against G− of 16.48% versus 16.66%. In addition to the strong antibacterial activity, this study confirmed the safety and quality of Czech honeys and helped to select the character of a possible source for in vivo testing and subsequent clinical trials

Antibiotic Resistance in Nosocomial Bacteria Isolated from Infected Wounds of Hospitalized Patients in Czech Republic

Hospitalized patients with wounds face an increased risk of infection with multi-drug-resistant nosocomial bacteria. In this study, samples from almost 10,000 patients from big hospitals in Czech Republic with infected wounds were analyzed for the presence of bacterial pathogens. In 7693 patients (78.8%), bacterial etiological agents were identified. Members of the Enterobacterales (37.1%) and Staphyloccus aureus (21.1%) were the most prevalent pathogens. Staphyloccus aureus showed methicillin resistance in 8.6%. Almost half of the Klebsiella pneumoniae isolates were ESBL-positive and 25.6% of the Enterobacter spp. isolates were AmpC-positive. The third most prevalent Pseudomonas aeruginosa showed resistance to 19&ndash;32% of the antipseudomonal antibiotics tested. Based on the results, amoxicillin/clavulanic acid, ampicillin/sulbactam or piperacillin/tazobactam combined with gentamicin can be recommended for antibiotic treatment of infected wounds. Once the etiological agent is identified, the therapy should be adjusted according to the species and its resistance