Inhibitory effect of selected medicinal plants on the release of pro-inflammatory cytokines in lipopolysaccharide-stimulated human peripheral blood mononuclear cells

The inhibitory activities of the methanol extracts from 20 selected medicinal plants on the release of pro-inflammatory cytokines in human peripheral blood mononuclear cells (PBMCs) were evaluated. The major compound from the most active plant extract was also investigated. The inhibitory effect of the methanol extracts on the release of pro-inflammatory cytokines was tested by incubating PBMCs with the sample and then stimulating by lipopolysaccharide at 0.1 μg/ml. The level of cytokines was determined using enzyme-linked immunosorbent assay. Among the extracts tested, Andrographis paniculata extract demonstrated the strongest inhibition of interleukin (IL)-1β, IL-1α, and IL-6 release, with IC₅₀ values of 1.54, 1.06, and 0.74 μg/ml, respectively. The IC₅₀ value of A. paniculata extract was significantly higher than that of andrographolide on IL-1α, IL-1β, and IL-6 (p < 0.001) release. The IC₅₀ values of andrographolide for IL-1α, IL-1β, and IL-6 were significantly higher (p < 0.001) than that of dexamethasone. Cymbopogon citratus and Zingiber officinale strongly inhibited the release of IL-1β, with IC₅₀ values of 3.22 and 3.17 μg/ml, respectively. To our knowledge, this is the first report that A. paniculata extract and its major compound andrographolide strongly inhibited the release of IL-1α, whereas previous studies only showed their inhibitory effect on the release of another IL-1 family member, IL-1β. The results show that these extracts and this compound have potential effects as anti-inflammatory agents by inhibiting the release of pro-inflammatory cytokines.     

Impact of a high-density grid catheter on long-term outcomes for structural heart disease ventricular tachycardia ablation

Journal of Interventional Cardiac Electrophysiology - Substrate mapping has highlighted the importance of targeting diastolic conduction channels and late potentials during ventricular tachycardia...     

Occupational leptospirosis in a Colombian Caribbean area

OBJECTIVE: To establish the seroprevalence of infection by Leptospira in an occupational setting in Cordoba. MATERIAL AND METHODS: A cross-sectional study was conducted among 334 farmworkers, butchers, and garbage collectors, to identify the presence of anti-leptospira IgM antibodies, in the Department of Cordoba, Colombia. Stratified sampling proportional to the number of inhabitants and occupation was used to select the sample population (confidence level 99.9%, error 0.5%, prevalence 72%).The SPSS software 11.0 version was used to perform non parametric tests with p < 0.05, as well as odds ratios with confidence intervals. RESULTS: The prevalence of previous infection by Leptospira was high (13.1%). No differences among areas were found, however, higher infection was associated with living in Cienaga de Oro municipality (OR = 3.52 Cl 1.70-7.26) (p = 0,00283). Being a farmer was also a risk factor for infection (OR = 2.04 Cl 1.080-3.85) (p = 0.025), as well as drinking water from a dam (OR = 2.4 CI 1.24-4.70) (p = 0.00787). CONCLUSIONS: The rate of infection is important and a significant public health problem in this area of the Colombian Caribbean coast.     

Insulin Regulates the Unfolded Protein Response in Human Adipose Tissue

Endoplasmic reticulum (ER) stress is increased in obesity and is postulated to be a major contributor to many obesity-related pathologies. Little is known about what causes ER stress in obese people. Here, we show that insulin upregulated the unfolded protein response (UPR), an adaptive reaction to ER stress, in vitro in 3T3-L1 adipocytes and in vivo, in subcutaneous (sc) adipose tissue of nondiabetic subjects, where it increased the UPR dose dependently over the entire physiologic insulin range (from ∼35 to ∼1,450 pmol/L). The insulin-induced UPR was not due to increased glucose uptake/metabolism and oxidative stress. It was associated, however, with increased protein synthesis, with accumulation of ubiquitination associated proteins, and with multiple posttranslational protein modifications (acetylations, methylations, nitrosylations, succinylation, and ubiquitinations), some of which are potential causes for ER stress. These results reveal a new physiologic role of insulin and provide a putative mechanism for the development of ER stress in obesity. They may also have clinical and therapeutic implications, e.g., in diabetic patients treated with high doses of insulin.     

Platelets are efficient and protective depots for storage,distribution, and delivery of lysosomal enzyme in mice with Hurler Syndrome

Use of megakaryocytes/platelets for transgene expression may take advantage of their rapid turnover and protective storage in platelets and reduce the risk of activating oncogenes in hematopoietic stem and progenitor cells (HSCs). Here, we show that human megakaryocytic cells could overexpress the lysosomal enzyme, α-l-iduronidase (IDUA), which is deficient in patients with mucopolysaccharidosis type I (MPS I). Upon megakaryocytic differentiation, the amount of released enzyme increased rapidly and steadily by 30-fold. Using a murine MPS I model, we demonstrated that megakaryocyte/platelets were capable of producing, packaging, and storing large amounts of IDUA with proper catalytic activity, lysosomal trafficking, and receptor-mediated uptake. IDUA can be released directly into extracellular space or within microparticles during megakaryocyte maturation or platelet activation, while retaining the capacity for cross-correction in patient’s cells. Gene transfer into 1.7% of HSCs led to long-term normalization of plasma IDUA and preferential distribution of enzyme in liver and spleen with complete metabolic correction in MPS I mice. Detection of GFP (coexpressed with IDUA) in Kupffer cells and hepatocytes suggested liver delivery of platelet-derived IDUA possibly via the clearance pathway for senile platelets. These findings provide proof of concept that cells from megakaryocytic lineage and platelets are capable of generating and storing fully functional lysosomal enzymes and can also lead to efficient delivery of both the enzymes released into the circulation and those protected within platelets/microparticles. This study opens a door for use of the megakaryocytes/platelets as a depot for efficient production, delivery, and effective tissue distribution of lysosomal enzymes.The potential of therapeutic benefits from genetically modified hematopoietic stem cells (HSCs) has been supported in recent gene therapy clinical trials (1, 2). High transgene dosage or selective growth of genetically corrected HSCs appears to be necessary for achieving clinical efficacy. However, genotoxic risk caused by proviral integration-associated oncogenesis is directly concomitant with high numbers of integration events or clonal expansion (3, 4). New approaches are needed to balance the need for high transgene frequency while limiting the associated increased risk of oncogenesis.Platelets are anuclear, secretory particulate entities containing proteins stored in cytoplasmic granules that can be released upon activation (5). Healthy adults produce 2–5 × 10¹¹ platelets daily with a baseline activation rate of 1–5% (6). Use of megakaryocytes (MKs)/platelets for transgene expression may (i) take advantage of this immense cell mass and its rapid turnover (5–9 d); (ii) provide protective storage of the transgene product, which is essential for proteins sensitive to plasma pH; and (iii) continuously dispense proteins via degranulation from platelet activation at baseline (without detectable injury) and/or at sites of vascular injury. Highly efficient protein production and delivery could further reduce the need for high transgene frequency and the risk of activating oncogenes in HSCs and all their progeny. Although using platelets as a delivery system has been demonstrated for the expression of coagulation factors to treat inherited bleeding disorders in mice (7, 8), there has been no report of the feasibility of using MKs/platelets for the generation of nonhematologic proteins.Lysosomal storage diseases (LSDs) are a group of inherited disorders, often affecting multiple organs including the liver and spleen, with a cumulative incidence of 1 in 5,000–7,000 live births (9). Overexpressing lysosomal enzymes in platelets not only can provide the protection of pH-sensitive enzymes and continuous enzyme release via low physiological levels of platelet activation but may also offer the benefit of on-target delivery of platelet-derived enzymes to spleen and liver in the process of platelet clearance (10). However, maintaining proper posttranslational modifications for appropriate lysosomal trafficking and intercellular lysosomal enzyme transfer is essential for metabolic cross correction in treating these multiorgan diseases (11). It is not known whether lysosomal enzymes generated from the MK/platelet lineage would be fully functional and capable of correcting lysosomal deficits in diseased cells.In this study, we used a mouse model of Hurler syndrome, which is the severe form of mucopolysaccharidosis type I (MPS I), one of most common LSDs. It is caused by the deficiency of α-l-iduronidase (IDUA) and consequent accumulation of glycosaminoglycans (GAGs) (12, 13). We show that MKs are capable of producing large amounts of IDUA with proper catalytic function, lysosomal trafficking and receptor-mediated uptake, which could be sorted to and stored within platelets. The IDUA can be released directly into the extracellular space or within microparticles (MPs) during MK maturation or platelet activation, while retaining its ability to cross-correct cells derived from patients with MPS I.