From Geraniums to Guadalajara on the Virtues of Early Retirement.

This is the story of author's life from high school through retirement. The author took early retirement after 25 years of practice in a multispecialty clinic then founded a free clinic in Mexico that the author ran for 15 years.

Direct Support Professional and Frontline Supervisor Perspectives on Work-Life in a Pandemic.

Direct support professionals (DSPs) and frontline supervisors (FLSs) play an invaluable role in delivering home and community-based services to people with intellectual and developmental disabilities (IDD). DSPs provide support with employment, community living, developing social connections, health and well-being, and many other areas. FLSs' primary responsibility is to provide direction and guidance to DSPs in their work; however, they also frequently provide direct support to people with IDD. This workforce has been in crisis with high turnover and vacancy which threaten the inclusion of individuals with disabilities in their communities. The COVID-19 pandemic exacerbated an already fraught situation.

Anti-inflammatory effects of natural product formulations on murine dendritic cells.

The popularity and availability of herbal extracts has increased dramatically over the last decade, providing an inexpensive manner of self-medication. Although the efficacy of individual extracts is currently being studied intensively, research regarding complex mixtures is limited. Therefore, we evaluated the effects of three complex formulations, including BRC-301, a polyherbal extract; BRC-304, a mixture of vitamins, minerals, antioxidant enzymes, botanical extracts, and carotenoids; and BRC-306, a proprietary blend of Uncaria tomentosa (cat's claw) and Phytolens(®) on murine dendritic cells (DCs). We hypothesized that these formulations would decrease the inflammatory responsiveness and innate function of DCs. In order to address this hypothesis, we evaluated the effects of BRC-301, BRC-304, and BRC-306 on DC2.4 cells and assessed the effects of BRC-301 on bone marrow-derived DCs (bmDCs). Lipopolysaccharide (LPS) stimulation of DC2.4 cells and bmDCs induced production of nitric oxide (NO), TNF-α, and IL-6, a response that was modulated by concomitant treatment with non-cytotoxic concentrations of BRC-301. In contrast, only the production of NOor IL-6 by LPS-activated DC2.4 cells was affected by BRC-304 or BRC-306, respectively. Flow cytometric evaluation following concurrent BRC-301 and LPS treatment revealed an increased relative expression of CD11c, CD86, and CD54 on bmDCs and an increased frequency of bmDCs expressing MHC II. Finally, BRC-301 enhanced the uptake of fluorescein isothiocyanate-conjugated ovalbumin by bmDCs. Taken together, these results suggest that these commercially available formulations modulate the innate responsiveness of murine DCs and may enhance their ability to initiate T cell-mediated immunity.

Interlaboratory comparison of three multiplexed bead-based immunoassays for measuring serum antibodies to pneumococcal polysaccharides.

Serotype-specific IgG, as quantified by a standardized WHO enzyme-linked immunosorbent assay (ELISA), is a serologic end point used to evaluate pneumococcal polysaccharide-based vaccine immunogenicity. Antibodies to each vaccine polysaccharide in licensed multivalent vaccines are quantified separately; this is laborious and consumes serum. We compared three bead-based immunoassays: a commercial assay (xMAP Pneumo14; Luminex) and two in-house assays (of the Health Protection Agency [HPA] and Centers for Disease Control and Prevention [CDC]), using the WHO-recommended standard reference and reference sera (n = 11) from vaccinated adults. Multiple comparisons of the IgG concentrations for seven conjugate vaccine serotypes were performed by sample (percent error), serotype (equivalency testing), and laboratory (concordance correlation coefficient [CCC]). When comparing concentrations by sample, bead-based immunoassays generally yielded higher antibody concentrations than the ELISA and had higher variability for serotypes 6B, 18C, and 23F. None of the three assays met the current WHO recommendation of 75% of sera falling within 40% of the assigned antibody concentrations for all seven serotypes. When compared by serotype, the CDC and HPA tests were equivalent for five of seven serotypes, whereas the Luminex assay was equivalent for four of seven serotypes. When overall mean IgG concentrations were compared by laboratory, a higher level of agreement (CCC close to 1) was found among bead-based immunoassays than between the assays and WHO assignments. When compared to WHO assignments, the HPA assay outperformed the other assays (r = 0.920; CCC = 0.894; coefficient of accuracy = 0.972). Additional testing with sera from immunogenicity studies should demonstrate the applicability of this methodology for vaccine evaluation.

Anti-inflammatory effects of natural product formulations on murine macrophages.

The popularity of herbal supplements, especially those with purported anti-inflammatory effects, has drastically increased in recent years as more people have turned to natural therapeutics. As the supplement industry is loosely regulated, the safety and efficacy of these products is poorly understood. In the present study, we examined the effects of natural product formulations prepared by the Biotics Research Corporation (BRC) on cyclooxygenase (COX) enzyme activity. We also evaluated the immune responsiveness of RAW264.7 macrophages, a key cell population involved in the inflammation, to those formulations. As a result, three supplements, BRC-301, BRC-304, and BRC-306, selectively inhibited COX-2, the inducible isoform involved in inflammation. Further evaluation of these three products indicated that BRC-304 and BRC-306 produced minimal effects on the production of inflammatory mediators by lipopolysaccharide (LPS)-stimulated macrophages. BRC-301 decreased the LPS-induced production of nitric oxide and IL-6, as well as CD40 expression. Collectively, these results suggest that the BRC-301 extract, comprising several polyphenolic natural products, may have a protective effect in chronic inflammatory disorders.

Evaluation of body mass index, pre-vaccination serum progesterone levels and anti-anthrax protective antigen immunoglobulin G on injection site adverse events following anthrax vaccination in women.

BACKGROUND: In 2002, CDC initiated the Anthrax Vaccination Program (AVP) to provide voluntary pre-exposure anthrax vaccination for individuals at high risk for exposure to Bacillus anthracis spores. The AVP offered an opportunity to investigate hypothesized reasons for a reported gender difference in injection site adverse events (AEs) following anthrax vaccine adsorbed (AVA). OBJECTIVES: To evaluate in women the impact of body mass index (BMI), pre-vaccination serum progesterone levels, and pre-vaccination anti-anthrax protective antigen immunoglobulin G concentrations (anti-PA IgG) on the occurrence of AEs following subcutaneous AVA vaccination. METHODS: Participants' BMI was determined at enrollment. Also, pre-vaccination blood samples were assayed for serum progesterone and anti-PA IgG. Post-vaccination solicited AEs were recorded by participants using a 4-day diary card. RESULTS: Obese group had an elevated risk for arm soreness. Decreased pre-vaccination serum progesterone level was associated with arm swelling. Increased pre-vaccination anti-PA IgG was associated with itching on the arm; and within the obese group, was associated with arm swelling, lump or knot, redness, soreness, and warmth. CONCLUSIONS: In AVA vaccinated women, obesity was associated with arm soreness and decreased pre-vaccination serum progesterone levels were associated with increased rate of arm swelling. Increased pre-vaccination anti-PA IgG may be associated with an increased frequency of itching on the arm, and in obese women, may increase the occurrence of arm swelling, lump or knot, redness, and warmth. Administering AVA according to a woman's menstrual phase may reduce the occurrence of certain injection site reactions.

Toll-like receptor ligand-induced activation of murine DC2.4 cells is attenuated by Panax notoginseng.

The medicinal herb, Panax notoginseng, has been used for thousands of years in traditional Chinese medicine and possesses anti-inflammatory properties. Dendritic cells (DCs) play a central role in the regulation of both inflammation and adaptive immunity. The aim of this study was to investigate the potential for notoginseng extracts to modulate Toll-like receptor (TLR) ligand-induced activation of cultured DC2.4 cells. Following stimulation with LPS, CpG or poly(I:C) and treatment with 0-50micorg/ml notoginseng extract for 24 h, DCs were evaluated for various phenotypic and functional readouts. Notoginseng reduced the LPS-, CpG- and poly(I:C)-induced production of TNF-alpha by DC2.4 cells. Also, IL-6 production by notoginseng-treated cells stimulated with LPS and CpG but not poly(I:C) was reduced when compared to controls. TLR ligand-induced CD40 expression was attenuated by notoginseng. In contrast, notoginseng decreased CD86 levels on DCs activated with LPS and poly(I:C) but not CpG. Inhibition of TNF-alpha production was time-dependent in LPS-stimulated cells, occurring only with pretreatment or concurrent treatment of notoginseng but not after delayed addition of the herbal extract. Additionally, ginsenoside Rg1 more effectively inhibited LPS-stimulated cytokine production by DC2.4 cells than ginsenoside Rb1. Taken together, these results demonstrate that notoginseng inhibits the production of specific inflammatory molecules and innate immune responsiveness by DCs following TLR activation.

Panax notoginseng attenuates LPS-induced pro-inflammatory mediators in RAW264.7 cells.

Herbals or dietary supplements are not regulated as drugs by the United States Food and Drug Administration (USFDA) although many may have associated therapeutic effects and toxicities. Therefore, the immunomodulatory effects of the herbal extract Panax notoginseng on cultured macrophages (RAW264.7 cells) were investigated to address potential therapeutic or toxic effects. Cells were stimulated with LPS (1 microg/ml) and treated with notoginseng at 5, 25 and 50 microg/ml. Notoginseng inhibited the LPS-induced production of TNF-alpha and IL-6 by the cultured macrophages in a concentration-dependent manner. The expression of COX-2 and IL-1 beta mRNA was also attenuated by notoginseng. TNF-alpha production was inhibited in samples treated with notoginseng 24h before, or at the same time as LPS stimulation, but not in samples treated 8h after LPS stimulation. Notoginseng reduced expression of the accessory molecules CD40 and CD86 on the RAW264.7 cells while CD14 and TLR4 expression remained unaffected. Furthermore, Rb1 and Rg1 ginsenosides also inhibited macrophage production of TNF-alpha, but to a lesser extent than did the whole notoginseng extract. Collectively, these results indicate that notoginseng inhibits LPS-induced activation of RAW264.7 macrophages and demonstrates that notoginseng possesses anti-inflammatory and immunosuppressive properties in vitro.

Differential activity of lipoic acid enantiomers in cell culture.

It is unclear whether the two enantiomeric forms (R & S) of lipoic acid (LA) share similar pharmacological activity and the exact cellular targets of LA are not well identified. We oxidatively stressed 3 cell culture systems representing different cell types. Mitochondrial metabolism was the primary endpoint. When C6 glioma was damaged by hydrogen peroxide (H2O2), all forms of LA protected. Racemic and S-LA were less effective than the R-isomer that was also protective in tertiary butyl hydroperoxide (TBHP)-damaged C6 glioma. In PC12 cells, little damage was produced by TBHP; R-LA increased mitochondrial metabolism above the level of non-damaged control. In H2O2 damaged PC12 cells, R-LA and racemic LA (but not S-LA) not only protected against damage, but increased mitochondrial metabolism above the non-damaged control level. When BAE cells were damaged with H2O2, R- and racemic LA protected while S-LA was ineffective.

Inhibition of COX isoforms by nutraceuticals.

Humans have two isoforms of Prostaglandin H Synthase or cyclooxygenase: COX-1 and COX-2. COX-1 is cytoprotective. COX-2 inhibitors reduce inflammation without the risk of ulceration and kidney damage. The ideal nutraceutical would inhibit COX-2 synthesis while preserving COX-1 synthesis. The hypothesis for this research was that COX inhibitors would fall primarily into three categories: COX-2 specific inhibition, non-specific inhibition (COX-1 and COX-2), and minimal inhibition. The human Cayman COX inhibitor screening assay was used to determine the inhibitory concentration 50 (IC50) of COX-1/ COX-2 activity of each nutraceutical. The assay was run, in duplicate, with three concentrations of a suspected inhibitor, a standard curve of eight concentrations, a non-specific binding sample, and a maximum binding sample. The inhibition and concentration of each sample was then put on a multiple regression best-fit line and the IC50 determined. For comparison, ibuprofen, rofecoxib, naproxen, and indomethacin were used. Positive results were seen for ipriflavone, resveratrol, MSV-60, amentoflavone, ruscus extract and notoginseng. Glucosamine, nexrutine, and berberine did not inhibit either isoform.