Elimination of Giardia duodenalis BIV in vivo using natural extracts in microbiome and dietary supplements

Abstract Background Prostate cancer (PCa) is a leading cause of cancer-related death in males worldwide. Exacerbated inflammation and angiogenesis have been largely demonstrated to contribute to PCa progression. Diverse naturally occurring compounds and dietary supplements are endowed with anti-oxidant, anti-inflammatory and anti-angiogenic activities, representing valid compounds to target the aberrant cytokine/chemokine production governing PCa progression and angiogenesis, in a chemopreventive setting. Using mass spectrometry analysis on serum samples of prostate cancer patients, we have previously found higher levels of carnitines in non-cancer individuals, suggesting a protective role. Here we investigated the ability of Acetyl-L-carnitine (ALCAR) to interfere with key functional properties of prostate cancer progression and angiogenesis in vitro and in vivo and identified target molecules modulated by ALCAR. Methods The chemopreventive/angiopreventive activities ALCAR were investigated in vitro on four different prostate cancer (PCa) cell lines (PC-3, DU-145, LNCaP, 22Rv1) and a benign prostatic hyperplasia (BPH) cell line. The effects of ALCAR on the induction of apoptosis and cell cycle arrest were investigated by flow cytometry (FC). Functional analysis of cell adhesion, migration and invasion (Boyden chambers) were performed. ALCAR modulation of surface antigen receptor (chemokines) and intracellular cytokine production was assessed by FC. The release of pro-angiogenic factors was detected by a multiplex immunoassay. The effects of ALCAR on PCa cell growth in vivo was investigated using tumour xenografts. Results We found that ALCAR reduces cell proliferation, induces apoptosis, hinders the production of pro inflammatory cytokines (TNF-α and IFN-γ) and of chemokines CCL2, CXCL12 and receptor CXCR4 involved in the chemotactic axis and impairs the adhesion, migration and invasion capabilities of PCa and BPH cells in vitro. ALCAR exerts angiopreventive activities on PCa by reducing production/release of pro angiogenic factors (VEGF, CXCL8, CCL2, angiogenin) and metalloprotease MMP-9. Exposure of endothelial cells to conditioned media from PCa cells, pre-treated with ALCAR, inhibited the expression of CXCR4, CXCR1, CXCR2 and CCR2 compared to those from untreated cells. Oral administration (drinking water) of ALCAR to mice xenografted with two different PCa cell lines, resulted in reduced tumour cell growth in vivo. Conclusions Our results highlight the capability of ALCAR to down-modulate growth, adhesion, migration and invasion of prostate cancer cells, by reducing the production of several crucial chemokines, cytokines and MMP9. ALCAR is a widely diffused dietary supplements and our findings provide a rational for studying ALCAR as a possible molecule for chemoprevention approaches in subjects at high risk to develop prostate cancer. We propose ALCAR as a new possible “repurposed agent’ for cancer prevention and interception, similar to aspirin, metformin or beta-blockers.

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Sublingual Reactivity to rBET V1 and rPHL P1 in Patients with Oral Allergy Syndrome

International Journal of Immunopathology and Pharmacology ◽

10.1177/205873920601900114 ◽

2006 ◽

Vol 19 (1) ◽

pp. 205873920601900 ◽

Cited By ~ 8

Author(s):

F. Marcucci ◽

L. Sensi ◽

G. DI Cara ◽

G. Gidaro ◽

C. Incorvaia ◽

...

Keyword(s):

Specific Ige ◽

Cross Reactivity ◽

Oral Allergy Syndrome ◽

Control Group ◽

Recombinant Allergens ◽

Natural Extracts ◽

Target Organs ◽

High Concentrations

Oral Allergy Syndrome (OAS) in patients with pollen-induced rhinoconjunctivitis is caused by specific IgE recognizing cross-reacting epitopes of fruits and plants, which were clearly shown in vitro, but failed to be demonstrated in vivo by cross-challenges in the target organs. Considering the hypothesis of degradation of such epitopes in natural extracts, challenges with recombinant pollen allergens were done to evaluate the reactivity of the oral mucosa in OAS patients. Seventeen patients with OAS and rhinitis from birch (10) and grass pollen (7) and 10 non-atopic controls were studied by skin prick tests (SPT), allergen specific nasal challenges (ASNC) and allergen specific sublingual challenges (ASSC) with birch and timothy extracts and with rBet v1 and rPhl p1 at increasing concentrations from 1 to 1000 mcg/ml. None of the healthy subjects in the control group had any positive test for birch and timothy extracts or for recombinant allergens. In the OAS group the following results were observed: SPTs with recombinant allergens were positive in all patients, mostly at 10 mcg/ml concentration; ASNC with rBet v1 were positive in all patients, mostly at 100 mcg/ml; ASSC with natural pollen extracts were positive in only 2 of 17 patients, but in 15 of 17 with rBet v1 and rPhl p1, mostly at 500 mcg/ml and 1000 mcg/ml. ASSC with rBet v1 and rPhl p1 were positive with a mean concentration of 677 and 533 mcg/ml, respectively. The results of sublingual challenges with rBet v1 and rPhl p1 showed the in vivo cross-reactivity between pollens and foods in patients with OAS, but high concentrations of the recombinant allergens were needed to reproduce oral symptoms, thus explaining the failure of challenges performed with natural extracts, which have concentrations of major allergens lower than 50 mcg/ml. This indicates that sublingual mucosa is much less reactive to allergens than other surfaces, such as skin and nasal mucosa, probably because of its anatomic and immunologic peculiarity.

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Giardia duodenalis-induced alterations of commensal bacteria killCaenorhabditis elegans: a new model to study microbial-microbial interactions in the gut

AJP Gastrointestinal and Liver Physiology ◽

10.1152/ajpgi.00335.2014 ◽

2015 ◽

Vol 308 (6) ◽

pp. G550-G561 ◽

Cited By ~ 31

Author(s):

Teklu K. Gerbaba ◽

Pratyush Gupta ◽

Kevin Rioux ◽

Dave Hansen ◽

Andre G. Buret

Keyword(s):

Bacterial Colonization ◽

Giardia Duodenalis ◽

Microbial Interactions ◽

Commensal Bacteria ◽

In Vivo Model ◽

Functional Changes ◽

E Coli ◽

C Elegans ◽

Human Microbiota

Giardia duodenalis is the most common cause of parasitic diarrhea worldwide and a well-established risk factor for postinfectious irritable bowel syndrome. We hypothesized that Giardia-induced disruptions in host-microbiota interactions may play a role in the pathogenesis of giardiasis and in postgiardiasis disease. Functional changes induced by Giardia in commensal bacteria and the resulting effects on Caenorhabditis elegans were determined. Although Giardia or bacteria alone did not affect worm viability, combining commensal Escherichia coli bacteria with Giardia became lethal to C. elegans. Giardia also induced killing of C. elegans with attenuated Citrobacter rodentium espF and map mutant strains, human microbiota from a healthy donor, and microbiota from inflamed colonic sites of ulcerative colitis patient. In contrast, combinations of Giardia with microbiota from noninflamed sites of the same patient allowed for worm survival. The synergistic lethal effects of Giardia and E. coli required the presence of live bacteria and were associated with the facilitation of bacterial colonization in the C. elegans intestine. Exposure to C. elegans and/or Giardia altered the expression of 172 genes in E. coli. The genes affected by Giardia included hydrogen sulfide biosynthesis (HSB) genes, and deletion of a positive regulator of HSB genes, cysB, was sufficient to kill C. elegans even in the absence of Giardia. Our findings indicate that Giardia induces functional changes in commensal bacteria, possibly making them opportunistic pathogens, and alters host-microbe homeostatic interactions. This report describes the use of a novel in vivo model to assess the toxicity of human microbiota.

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Giardia duodenalis induces paracellular bacterial translocation and causes postinfectious visceral hypersensitivity

AJP Gastrointestinal and Liver Physiology ◽

10.1152/ajpgi.00144.2015 ◽

2016 ◽

Vol 310 (8) ◽

pp. G574-G585 ◽

Cited By ~ 28

Author(s):

Marie C. M. Halliez ◽

Jean-Paul Motta ◽

Troy D. Feener ◽

Gaetan Guérin ◽

Laetitia LeGoff ◽

...

Keyword(s):

Giardia Duodenalis ◽

Intestinal Barrier ◽

Gastrointestinal Disorder ◽

Intraepithelial Lymphocytes ◽

Visceral Hypersensitivity ◽

New Model ◽

Infectious Gastroenteritis ◽

Irritable Bowel

Irritable bowel syndrome (IBS) is the most frequent functional gastrointestinal disorder. It is characterized by abdominal hypersensitivity, leading to discomfort and pain, as well as altered bowel habits. While it is common for IBS to develop following the resolution of infectious gastroenteritis [then termed postinfectious IBS (PI-IBS)], the mechanisms remain incompletely understood. Giardia duodenalis is a cosmopolitan water-borne enteropathogen that causes intestinal malabsorption, diarrhea, and postinfectious complications. Cause-and-effect studies using a human enteropathogen to help investigate the mechanisms of PI-IBS are sorely lacking. In an attempt to establish causality between giardiasis and postinfectious visceral hypersensitivity, this study describes a new model of PI-IBS in neonatal rats infected with G. duodenalis. At 50 days postinfection with G. duodenalis (assemblage A or B), long after the parasite was cleared, rats developed visceral hypersensitivity to luminal balloon distension in the jejunum and rectum, activation of the nociceptive signaling pathway (increased c- fos expression), histological modifications (villus atrophy and crypt hyperplasia), and proliferation of mucosal intraepithelial lymphocytes and mast cells in the jejunum, but not in the rectum. G. duodenalis infection also disrupted the intestinal barrier, in vivo and in vitro, which in turn promoted the translocation of commensal bacteria. Giardia-induced bacterial paracellular translocation in vitro correlated with degradation of the tight junction proteins occludin and claudin-4. The extensive observations associated with gut hypersensitivity described here demonstrate that, indeed, in this new model of postgiardiasis IBS, alterations to the gut mucosa and c- fos are consistent with those associated with PI-IBS and, hence, offer avenues for new mechanistic research in the field.

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Differential effectiveness of berry polyphenols as anti-giardial agents

Parasitology ◽

10.1017/s0031182011000825 ◽

2011 ◽

Vol 138 (9) ◽

pp. 1110-1116 ◽

Cited By ~ 7

Author(s):

J.-P. ANTHONY ◽

L. FYFE ◽

D. STEWART ◽

G. J. McDOUGALL

Keyword(s):

Intestinal Parasite ◽

Solid Phase ◽

Giardia Duodenalis ◽

Phase Extraction ◽

Coumaric Acid ◽

Phenol Content ◽

Differential Effectiveness ◽

Berry Extracts

SUMMARYFollowing previous work on the anti-giardial effect of blueberry polyphenols, a range of polyphenol-rich extracts from berries and other fruits was screened for their ability to kill Giardia duodenalis, an intestinal parasite of humans. Polyphenol-rich extracts were prepared from berries using solid-phase extraction and applied to trophozoites of Giardia duodenalis grown in vitro. All berry extracts caused inhibition at 166 μg gallic acid equivalents (GAE)/ml phenol content but extracts from strawberry, arctic bramble, blackberry and cloudberry were as effective as the currently used drug, metronidazole, causing complete trophozoite mortality in vitro. Cloudberry extracts were found to be the most effective causing effectively complete trophozoite mortality at 66 μg GAE/ml. The polyphenol composition of the more effective berry extracts suggested that the presence of ellagitannins could be an important factor. However, the potency of cloudberry could be related to high ellagitannin content but also to the presence of substantial amounts of unconjugated p-coumaric acid and benzoic acid. These in vitro effects occur at concentrations easily achievable in the gut after berry ingestion and we discuss the likelihood that berry extracts could be effective anti-giardial agents in vivo.

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The Multitarget Activity of Natural Extracts on Cancer: Synergy and Xenohormesis

Medicines ◽

10.3390/medicines6010006 ◽

2018 ◽

Vol 6 (1) ◽

pp. 6 ◽

Cited By ~ 7

Author(s):

María Herranz-López ◽

María Losada-Echeberría ◽

Enrique Barrajón-Catalán

Keyword(s):

Antitumor Agents ◽

Natural Extracts ◽

Potential Source ◽

Cancer Models ◽

Pure Compounds ◽

Synergistic Behavior ◽

Approved Drugs ◽

Initial Research

It is estimated that over 60% of the approved drugs and new drug developments for cancer and infectious diseases are from natural origin. The use of natural compounds as a potential source of antitumor agents has been deeply studied in many cancer models, both in vitro and in vivo. Most of the Western medicine studies are based on the use of highly selective pure compounds with strong specificity for their targets such as colchicine or taxol. Nevertheless, approximately 60% of fairly specific drugs in their initial research fail because of toxicity or ineffectiveness in late-stage preclinical studies. Moreover, cancer is a multifaceted disease that in most cases deserves a polypharmacological therapeutic approach. Complex plant-derived mixtures such as natural extracts are difficult to characterize and hardly exhibit high pharmacological potency. However, in some cases, these may provide an advantage due to their multitargeted mode of action and potential synergistic behavior. The polypharmacology approach appears to be a plausible explanation for the multigargeted mechanism of complex natural extracts on different proteins within the same signalling pathway and in several biochemical pathways at once. This review focuses on the different aspects of natural extracts in the context of anticancer activity drug development, with special attention to synergy studies and xenohormesis.

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Proteome and Transcriptome Analysis of the Antioxidant Mechanism in Chicken Regulated by Eucalyptus Leaf Polyphenols Extract

Oxidative Medicine and Cellular Longevity ◽

10.1155/2020/1384907 ◽

2020 ◽

Vol 2020 ◽

pp. 1-14

Author(s):

Wei Li ◽

Ze-qi He ◽

Xiao-Ying Zhang ◽

Yun-Jiao Chen ◽

Jian-Jun Zuo ◽

...

Keyword(s):

Antioxidant Capacity ◽

Dietary Supplements ◽

Control Diet ◽

Breast Muscle ◽

Glutathione Metabolism ◽

Control Group ◽

Treatment Groups ◽

Antioxidant Mechanism ◽

Mda Content

Eucalyptus leaf polyphenols extract (EPE) has been proved to have various bioactivities, but few reports focus on its antioxidant mechanism in vivo. The purpose of this study was to elucidate the effect and mechanism of EPE dietary supplements on antioxidant capacity in chicken. A total of 216 chickens were randomly selected for a 40-day experiment. Four treatment groups received diets including the control diet only, the control diet + low EPE (0.6 g/kg), the control diet + moderate EPE (0.9 g/kg), and the control diet + high EPE (1.2 g/kg). Compared with control group, the glutathione peroxidase (GSH-Px) activity and glutathione (GSH) content in the breast muscle of the moderate EPE treatment group was significantly higher (p<0.05), while the malonaldehyde (MDA) content in the moderate EPE group was reduced (p<0.05). Moreover, proteomic and transcriptomic analyses of the breast muscle revealed that glutathione metabolism and the peroxisome were the two crucial metabolic pathways responsible for increased antioxidant capacity of the muscle. Accordingly, nine candidate genes and two candidate proteins were identified related to improved antioxidant status induced by EPE supplements. This research provides new insights into the molecular mechanism of antioxidant capacity in chickens treated with EPE dietary supplements.

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The 3Rs: Reduction and refinement through a multivariate statistical analysis approach in a behavioural study to unveil anxiolytic effects of natural extracts of Tilia tomentosa

Biomedical Science and Engineering ◽

10.4081/bse.2019.116 ◽

2020 ◽

Cited By ~ 1

Author(s):

Guendalina Olivero ◽

Federica Turrini ◽

Matteo Vergassola ◽

Raffaella Boggia ◽

Paola Zunin ◽

...

Keyword(s):

Statistical Approach ◽

Principal Component ◽

Experimental Protocol ◽

Multivariate Statistical ◽

Data Set ◽

Natural Extracts ◽

Multivariate Statistical Approach ◽

Behavioural Tests ◽

Tilia Tomentosa

We propose a multivariate statistical approach based on Principal Component Analysis (PCA) as an useful instrument to improve the Rules of Refinement and Reduction in in vivo animal experimentation. We analysed with PCA the preliminary data from a study on the effects of the oral administration of Tilia tomentosa bud extracts (TTBEs) on the behavioural skills of adult and aged male and female mice. PCA allows to rationalize the data set information and to dissect the results, showing connections among variables under study (behavioural parameters) and different trends in the experimental groups (control and TTBEs-administered animals). Our results show that PCA can give some important information that can be useful for the refinement of the experimental protocol, in order to reduce the number of the animals used in the experiments and/or the behavioural tests to get reliable information.

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Zoonotic Parasitological Findings in a Puppy: The Course and Therapeutical Efficacy

Folia Veterinaria ◽

10.2478/fv-2021-0018 ◽

2021 ◽

Vol 65 (2) ◽

pp. 58-67

Author(s):

Ľ. Burcáková ◽

G. Štrkolcová ◽

A. Königová ◽

M. Várady

Keyword(s):

Close Contact ◽

Companion Animals ◽

Giardia Duodenalis ◽

Toxocara Canis ◽

Experimental Therapy ◽

Dipylidium Caninum ◽

Faecal Egg Count ◽

Before And After ◽

Parasitic Zoonoses

Abstract Considering the close contact of companion animals and humans, gastrointestinal parasitic zoonoses are very widespread and represent a high risk of transmission with the potential of severe consequences affecting the digestive tract of both humans and other animals. In this study we focused on enteric zoonoses caused by Toxocara canis nematode, Dipylidium caninum tapeworm and Giardia duodenalis protozoa. Our primary aim was to observe Toxocara canis egg excretion within the 27 consecutive days before and after orally treatment (2 Caniverm® tablets) on Day 13 in a naturally infected puppy. An average egg per gram (EPG) of T. canis detected by coprological quantitative McMaster method was 4558.33 and 666.66, before and after treatment, respectively. The percentage of faecal egg count reduction (%FECR) in in vivo Faecal Egg Count Reduction Test (FECRT) has confirmed an 85.37 % efficacy against T. canis. Secondly, the efficacy of Caniverm® against the tapeworm Dipylidium caninum was also determined. No D. caninum proglottides were detected on Day 14. The data showed 100 % effectiveness of the anthelmintic treatment. Metrobactin® 250 mg has been tested as experimental therapy against Giardia duodenalis on Day 3. On day 10, no cysts were observed in the faeces after per os ¼ tablet administration twice a day for 7 days.

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