Regulatory T Cells, a Potent Immunoregulatory Target for CAM Researchers: Modulating Tumor Immunity, Autoimmunity and Alloreactive Immunity (III)

Regulatory T (T(reg)) cells are the major arbiter of immune responses, mediating actions through the suppression of inflammatory and destructive immune reactions. Inappropriate T(reg) cell frequency or functionality potentiates the pathogenesis of myriad diseases with ranging magnitudes of severity. Lack of suppressive capability hinders restraint on immune responses involved in autoimmunity and alloreactivity, while excessive suppressive capacity effectively blocks processes necessary for tumor destruction. Although the etiology of dysfunctional T(reg) cell populations is under debate, the ramifications, and their mechanisms, are increasingly brought to light in the medical community. Methods that compensate for aberrant immune regulation may not address the underlying complications; however, they hold promise for the alleviation of debilitating immune system-related disorders. The dominant immunoregulatory nature of T(reg) cells, coupled with recent mechanistic knowledge of natural immunomodulatory compounds, highlights the importance of T(reg) cells to practitioners and researchers of complementary and alternative medicine (CAM)

Regulatory T Cells, a Potent Immunoregulatory Target for CAM Researchers: Modulating Allergic and Infectious Disease Pathology (II)

Regulatory T (T(reg)) cells maintain dominant control of immune responses to foreign materials and microbes. Appropriate T(reg) cell suppression of immune responses is essential for the maintenance of efficacious defensive responses and the limitation of collateral tissue damage due to excess inflammation. Allergy and infection are well studied and frequent afflictions in which T(reg) cells play an essential role. As such, they provide excellent models to communicate the significance and relevance of T(reg) cells to complementary and alternative medicine (CAM)

Regulatory T Cells, a Potent Immunoregulatory Target for CAM Researchers: The Ultimate Antagonist (I)

Over the past decade, great interest has been given to regulatory T (T(reg)) cells. A vast body of evidence has shown the existence and highlighted the importance of T(reg) cells in the active suppression of immune system responses. This form of immunoregulation is the dominant means utilized by the immune system to reach a harmony between reciprocal response processes in order to ensure adequate host defense with minimal host detriment. Therapeutically targeting T(reg) cells is a direct and powerful means to manipulate the immune system to achieve beneficial effects on various disease pathologies, including allergy, autoimmunity and cancer, as well as the facilitation of organ transplantation. This powerful target for immunoregulation is of much concern to practitioners and researchers of complementary and alternative medicine because it allows a great deal of control and certainty in dealing with the prevalence of debilitating immune system-related disorders for which there has been little remedy outside of Western Medicine

Pyrophosphate-Dependent ATP Formation from Acetyl Coenzyme A in Syntrophus aciditrophicus, a New Twist on ATP Formation.

UnlabelledSyntrophus aciditrophicus is a model syntrophic bacterium that degrades key intermediates in anaerobic decomposition, such as benzoate, cyclohexane-1-carboxylate, and certain fatty acids, to acetate when grown with hydrogen-/formate-consuming microorganisms. ATP formation coupled to acetate production is the main source for energy conservation by S. aciditrophicus However, the absence of homologs for phosphate acetyltransferase and acetate kinase in the genome of S. aciditrophicus leaves it unclear as to how ATP is formed, as most fermentative bacteria rely on these two enzymes to synthesize ATP from acetyl coenzyme A (CoA) and phosphate. Here, we combine transcriptomic, proteomic, metabolite, and enzymatic approaches to show that S. aciditrophicus uses AMP-forming, acetyl-CoA synthetase (Acs1) for ATP synthesis from acetyl-CoA. acs1 mRNA and Acs1 were abundant in transcriptomes and proteomes, respectively, of S. aciditrophicus grown in pure culture and coculture. Cell extracts of S. aciditrophicus had low or undetectable acetate kinase and phosphate acetyltransferase activities but had high acetyl-CoA synthetase activity under all growth conditions tested. Both Acs1 purified from S. aciditrophicus and recombinantly produced Acs1 catalyzed ATP and acetate formation from acetyl-CoA, AMP, and pyrophosphate. High pyrophosphate levels and a high AMP-to-ATP ratio (5.9 ± 1.4) in S. aciditrophicus cells support the operation of Acs1 in the acetate-forming direction. Thus, S. aciditrophicus has a unique approach to conserve energy involving pyrophosphate, AMP, acetyl-CoA, and an AMP-forming, acetyl-CoA synthetase.ImportanceBacteria use two enzymes, phosphate acetyltransferase and acetate kinase, to make ATP from acetyl-CoA, while acetate-forming archaea use a single enzyme, an ADP-forming, acetyl-CoA synthetase, to synthesize ATP and acetate from acetyl-CoA. Syntrophus aciditrophicus apparently relies on a different approach to conserve energy during acetyl-CoA metabolism, as its genome does not have homologs to the genes for phosphate acetyltransferase and acetate kinase. Here, we show that S. aciditrophicus uses an alternative approach, an AMP-forming, acetyl-CoA synthetase, to make ATP from acetyl-CoA. AMP-forming, acetyl-CoA synthetases were previously thought to function only in the activation of acetate to acetyl-CoA

Novel Diagnosis of Lyme Disease: Potential for CAM Intervention

Lyme disease (LD) is the most common tick-borne disease in the northern hemisphere, producing a wide range of disabling effects on multiple human targets, including the skin, the nervous system, the joints and the heart. Insufficient clinical diagnostic methods, the necessity for prompt antibiotic treatment along with the pervasive nature of infection impel the development and establishment of new clinical diagnostic tools with increased accuracy, sensitivity and specificity. The goal of this article is 4-fold: (i) to detail LD infection and pathology, (ii) to review prevalent diagnostic methods, emphasizing inherent problems, (iii) to introduce the usage of in vivo induced antigen technology (IVIAT) in clinical diagnostics and (iv) to underscore the relevance of a novel comprehensive LD diagnostic approach to practitioners of Complementary and Alternative Medicine (CAM). Utilization of this analytical method will increase the accuracy of the diagnostic process and abridge the time to treatment, with antibiotics, herbal medicines and nutritional supplements, resulting in improved quality of care and disease prognosis

High throughput analysis of proteome perturbations induced by radiation, radiomitigators and chemotherapeutics

Proteomics has flourished as a result of major advances in computational power, data storage technologies, and network architecture, all of which enable the acquisition, handling, and processing of enormous data sets. As mass spectrometers continue to break resolution and sensitivity barriers, the limitations of qualitative and quantitative proteomic experiments will no longer result from instrumentation constraints, but rather from the quality and availability of robust sample preparation methods and post-processing algorithms. My dissertation focuses on the development of technologies in experimental proteomic workflows that precede and that follow mass spectrometric analysis - areas that are integral to the future of proteomics. Enhanced Filter Aided Sample Preparation (eFASP) incorporates Tween-20 for plastic passivation, and n-octylglucoside and deoxycholic acid for digestion enhancement. In E. coli, eFASP reduces sample losses associated with normal FASP by almost 300%, and significantly increases hydrophobic protein representation when compared to normal FASP and to a routine precipitation-based sample preparation. eFASP is validated in the analysis of the ultra-stable proteinaceous sheath of Methanospirillum hungatei. eFASP and label-free quantitative analysis revealed a 40.7 kDa polypeptide (Q2FRN9) encoded by Mhun_2271 as the predominant sheath component by 10-fold. eFASP is combined with sample fractionation methods and software tools for the quantitative analysis of bacterial and human samples. Hydrophilic interaction chromatography (HILIC) enables small-scale fractionation for less complex samples: Syntrophus aciditrophicus, Anaerobaculum hydrogeniformans, and melanoma-derived exosomes. S. aciditrophicus and A. hydrogeniformans are profiled for proteome shifts resulting changes in carbon source. Treatment of melanoma cells with B-Raf inhibitor, PLX-4720, results in the packaging of two tyrosine kinases, VEGFR and PDGFR-B, into secreted exosomes. Upregulation of these receptor tyrosine kinases has been found to be a result of the release of feedback inhibition following treatment with MAPK targeted therapeutics. Complex whole-cell proteome analysis is effected by high-resolution electrostatic-repulsion hydrophilic interaction chromatography (ERLIC) fractionation of eFASP-processed samples. eFASP-ERLIC facilitated the quantitative analysis of normal and ATM-deficient human lymphoblastoid cell lines (LCLs) exposed to ionizing radiation (IR) and to YEL2, a potent radiomitigator. Exposure to IR resulted in the durable upregulation of NRF2-oxidative stress response components in Normal LCLs, while thrombin signaling and pro-survival proteins were upregulated in ATM-deficient LCLs. YEL2 treatment upregulated DNA repair components and enriched pro-survival pathways, which implicate the involvement of multiple processes in YEL2-mediated radioresistance