Cinnamic Acid Bornyl Ester Derivatives from Valeriana wallichii Exhibit Antileishmanial In Vivo Activity in Leishmania major-Infected BALB/c Mice

Human leishmaniasis covers a broad spectrum of clinical manifestations ranging from selfhealing cutaneous leishmaniasis to severe and lethal visceral leishmaniasis caused among other species by Leishmania major or Leishmania donovani, respectively. Some drug candidates are in clinical trials to substitute current therapies, which are facing emerging drugresistance accompanied with serious side effects. Here, two cinnamic acid bornyl ester derivatives (1 and 2) were assessed for their antileishmanial activity. Good selectivity and antileishmanial activity of bornyl 3-phenylpropanoate (2) in vitro prompted the antileishmanial assessment in vivo. For this purpose, BALB/c mice were infected with Leishmania major promastigotes and treated with three doses of 50 mg/kg/day of compound 2. The treatment prevented the characteristic swelling at the site of infection and correlated with reduced parasite burden. Transmitted light microscopy and transmission electron microscopy of Leishmania major promastigotes revealed that compounds 1 and 2 induce mitochondrial swelling. Subsequent studies on Leishmania major promastigotes showed the loss of mitochondrial transmembrane potential (ΔΨm) as a putative mode of action. As the cinnamic acid bornyl ester derivatives 1 and 2 had exhibited antileishmanial activity in vitro, and compound 2 in Leishmania major-infected BALB/c mice in vivo, they can be regarded as possible lead structures for the development of new antileishmanial therapeutic approaches

Antileishmanial Lead Structures from Nature: Analysis of Structure-Activity Relationships of a Compound Library Derived from Caffeic Acid Bornyl Ester

Bioassay-guided fractionation of a chloroform extract of Valeriana wallichii (V. wallichii) rhizomes lead to the isolation and identification of caffeic acid bornyl ester (1) as the active component against Leishmania major (L. major) promastigotes (IC50 = 48.8 µM). To investigate the structure-activity relationship (SAR), a library of compounds based on 1 was synthesized and tested in vitro against L. major and L. donovani promastigotes, and L. major amastigotes. Cytotoxicity was determined using a murine J774.1 cell line and bone marrow derived macrophages (BMDM). Some compounds showed antileishmanial activity in the concentration range of pentamidine and miltefosine which are the standard drugs in use. In the L. major amastigote assay compounds 15, 19 and 20 showed good activity with relatively low cytotoxicity against BMDM, resulting in acceptable selectivity indices. Molecules with adjacent phenolic hydroxyl groups exhibited elevated cytotoxicity against murine cell lines J774.1 and BMDM. The Michael system seems not to be essential for antileishmanial activity. Based on the results compound 27 can be regarded as new lead structure for further structure optimizatio

Cinnamic Acid Bornyl Ester Derivatives from <i>Valeriana wallichii</i> Exhibit Antileishmanial <i>In Vivo</i> Activity in <i>Leishmania major</i>-Infected BALB/c Mice

<div><p>Human leishmaniasis covers a broad spectrum of clinical manifestations ranging from self-healing cutaneous leishmaniasis to severe and lethal visceral leishmaniasis caused among other species by <i>Leishmania major</i> or <i>Leishmania donovani</i>, respectively. Some drug candidates are in clinical trials to substitute current therapies, which are facing emerging drug-resistance accompanied with serious side effects. Here, two cinnamic acid bornyl ester derivatives (<b>1</b> and <b>2</b>) were assessed for their antileishmanial activity. Good selectivity and antileishmanial activity of bornyl 3-phenylpropanoate (<b>2</b>) <i>in vitro</i> prompted the antileishmanial assessment <i>in vivo</i>. For this purpose, BALB/c mice were infected with <i>Leishmania major</i> promastigotes and treated with three doses of 50 mg/kg/day of compound <b>2</b>. The treatment prevented the characteristic swelling at the site of infection and correlated with reduced parasite burden. Transmitted light microscopy and transmission electron microscopy of <i>Leishmania major</i> promastigotes revealed that compounds <b>1</b> and <b>2</b> induce mitochondrial swelling. Subsequent studies on <i>Leishmania major</i> promastigotes showed the loss of mitochondrial transmembrane potential (ΔΨ_m) as a putative mode of action. As the cinnamic acid bornyl ester derivatives <b>1</b> and <b>2</b> had exhibited antileishmanial activity <i>in vitro</i>, and compound <b>2</b> in <i>Leishmania major</i>-infected BALB/c mice <i>in vivo</i>, they can be regarded as possible lead structures for the development of new antileishmanial therapeutic approaches.</p></div

A comparative study of lipid profile and autonomic functions in vegetarian and non-vegetarian postmenopausal women

Background: The prevalence of dyslipedaemia, autonomic dysfunction leading to cardiovascular diseases, increases with menopause and an ageing population. Autonomic dysfunction as measured by lower heart rate variability is an established risk factor for cardiac death. Diet and nutrition have been extensively investigated as risk factors for major cardiovascular diseases and are also linked to other cardiovascular risk factors. Objectives: To compare lipid profile and autonomic functions of postmenopausal women on vegetarian and non- vegetarian diet. Materials and Methods: 120 Postmenopausal women (menopausal duration and age-matched) without any gross systemic disease from an Industrial population were selected. Sixty women were on vegetarian diet and 60 on non-vegetarian diet. BMI and waist/hip ratios were calculated, lipid profile was analyzed, and autonomic function tests were carried out. A comparison was done between the two groups using Students t test. Pearson′s correlation coefficient was calculated between the independent variable (lipid profile parameters) and the dependent variables (deep breath test, valsalva ratio, 30:15 ratio, OTT, IHG, CPT) to understand the effect of lipid profile on autonomic control of heart. Results : Significant increases in total cholesterol, triglyceride, LDL, cholesterol/HDL ratio were noticed in women on non-vegetarian diet. Results of autonomic function tests, i.e. valsalva ratio, deep breath test, 30: 15R-R intervals ratio, isometric hand grip test, cold pressor test, and orthostatic tolerance test were significantly worsened in postmenopausal women on non-vegetarian diet. Conclusion: Dietary factors may be an important cause of alteration of lipid metabolism. Increased cholesterol decreases heart rate variability and increased LDL cholesterol decreases baroreceptor sensitivity thereby worsening autonomic functions in postmenopausal women

Changes in cell morphology in <i>L</i>. <i>major</i> promastigotes upon treatment.

<p>Compound 1 and 2 induce parasite swelling and vacuolization of <i>L</i>. <i>major</i> promastigotes at very early time points. <i>L</i>. <i>major</i> promastigotes were treated with solvent control (1% DMSO), miltefosine (122.7 μM), compound 1 (100 μM), and 2 (100 μM) for 30 min, 1 h, 2 h, 4 h, and 24 h. Cytospin preparations of the cells were stained with Diff-Quik dye and analyzed by transmitted light microscopy under 50× objective. “Plus” symbol (+) was used to represent changes in morphology; two (++) and three (+++) symbols were used as a reference for the severity of the phenotype induced by the different tested compounds and the black arrows to indicate the presence of vacuole-like structures. Black bar indicates 20 μm.</p

Determination of cytotoxicity and IC₅₀ values against <i>Leishmania</i> parasites.

<p>Determination of cytotoxicity and IC₅₀ values against <i>Leishmania</i> parasites.</p

Compound-associated cell death is time-dependent.

<p><i>L</i>. <i>major</i> promastigotes were treated with solvent control (1% DMSO), apoptosis inducer miltefosine (122.7 μM), compound 1 (100 μM), and 2 (100 μM) in a time course experiment for 6 h, 10 h, and 24 h. Cells were harvested, stained with AV and PI and subsequently analyzed by flow cytometry. Analysis for compound 2 was performed in an independent experiment. Percentage for each quadrant is written below the corresponding dot blot figure. Lower-left (LL): live cells (AV^-/PI^-); upper-left (UL): early necrotic cells (AV^-/PI⁺); upper-right (UR): necrotic/late apoptotic cells (AV⁺/PI⁺); and lower-right (LR): early apoptotic cells (AV⁺/PI^-).</p

Global transcriptome analysis reveals fungal disease responsive core gene regulatory landscape in tea

Abstract Fungal infections are the inevitable limiting factor for productivity of tea. Transcriptome reprogramming recruits multiple regulatory pathways during pathogen infection. A comprehensive meta-analysis was performed utilizing previously reported, well-replicated transcriptomic datasets from seven fungal diseases of tea. The study identified a cumulative set of 18,517 differentially expressed genes (DEGs) in tea, implicated in several functional clusters, including the MAPK signaling pathway, transcriptional regulation, and the biosynthesis of phenylpropanoids. Gene set enrichment analyses under each pathogen stress elucidated that DEGs were involved in ethylene metabolism, secondary metabolism, receptor kinase activity, and various reactive oxygen species detoxification enzyme activities. Expressional fold change of combined datasets highlighting 2258 meta-DEGs shared a common transcriptomic response upon fungal stress in tea. Pervasive duplication events caused biotic stress-responsive core DEGs to appear in multiple copies throughout the tea genome. The co-expression network of meta-DEGs in multiple modules demonstrated the coordination of appropriate pathways, most of which involved cell wall organization. The functional coordination was controlled by a number of hub genes and miRNAs, leading to pathogenic resistance or susceptibility. This first-of-its-kind meta-analysis of host–pathogen interaction generated consensus candidate loci as molecular signatures, which can be associated with future resistance breeding programs in tea

Compound 2 treatment of <i>L</i>. <i>major</i>-infected BALB/c mice is associated with reduced parasite burden in infected tissues and organs.

<p>Female BALB/c mice (n = 33) were infected with 5 × 10⁶<i>L</i>. <i>major</i> promastigotes into the right hind footpad. Treatment regimen started on d 21 post infection either using 50% DMSO/PBS (□) (n = 11), 50 mg/kg compound 2 (■) (n = 11) or mice were left untreated (▲) (n = 11). A. At the end of experiment on d 35 all mice (n = 33) were sacrificed and the parasite burden of the infected footpads (iFP) (n = 9) was determined using Limited Dilution Assays (LDA). B. Photographic documentation of the infected footpads of three exemplary mice per group shall reveal the severity of disease progression in non-healing mice and the control of clinical manifestation in compound 2-treated BALB/c mice. C. The parasite burden of the infection site-draining popliteal lymph nodes (pLN) of individual mice (n = 9) or D. spleens of individual mice (n = 6) was determined using LDA. n = number of animals; ** p<0.005; differences in parasite load between 50% DMSO/PBS-treated and compound 2-treated animals did not reach statistical significance for D.</p