Capsaicin potently blocks Salmonella typhimurium invasion of Vero cells

As at 2021, the center for disease control (CDC) reported that Salmonella causes 1.2 million illness in the United States each year, with a mortality rate approaching 500 deaths per annum. Infants, the elderly, and persons with compromised immunity are the population with higher risk of mortality from this infection. At present there is no commercially available, safe and efficacious vaccine for the control and management of Salmonella typhimurium (S. typhimurium). More so, S. typhimurium has been shown to develop resistance against most antibiotics used for treatment of the infection. Capsaicin, a bioactive compound from Capsicum chinense (C. chinenses) is undoubtedly one of the most widely used spice in the world. This heat producing compound is not only been used as food additive but have been demonstrated to possess unique properties that have pharmacological, physiological, and antimicrobial applications. In this work, the antimicrobial property of pure capsaicin or capsaicin extract against S. typhimurium is tested to determine the compounds effectiveness in S. typhimurium inhibition. Capsaicin extract showed potent inhibition of S. typhimurium growth at concentrations as low as 100 ng/ml, whereas pure capsaicin comparatively showed poorer inhibition of the bacteria. Furthermore, both capsaicin extract and pure capsaicin potently blocked S. typhimurium invasion of an animal cell line in vitro. Taken together, this work revealed that capsaicin might work synergistically with dihydrocapsaicin or the other capsaicinoids to inhibit S. typhimurium growth, whereas individually, capsaicin or dihydrocapsaicin could potently block the bacteria entry and invasion of Vero cells.

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Antimicrobial Activity of Artemisinin and Precursor Derived fromIn VitroPlantlets ofArtemisia annuaL.

BioMed Research International ◽

10.1155/2014/215872 ◽

2014 ◽

Vol 2014 ◽

pp. 1-6 ◽

Cited By ~ 23

Author(s):

Suganthi Appalasamy ◽

Kiah Yann Lo ◽

Song Jin Ch'ng ◽

Ku Nornadia ◽

Ahmad Sofiman Othman ◽

...

Keyword(s):

Antimicrobial Activity ◽

Artemisia Annua ◽

Antimicrobial Property ◽

Bioactive Compound ◽

Antimicrobial Activities ◽

Hot Climate ◽

Inhibition Concentration ◽

Brine Shrimps ◽

Layer Chromatography

Artemisia annuaL., a medicinal herb, produces secondary metabolites with antimicrobial property. In Malaysia due to the tropical hot climate,A. annuacould not be planted for production of artemisinin, the main bioactive compound. In this study, the leaves of threein vitro A. annuaL. clones were, extracted and two bioactive compounds, artemisinin and a precursor, were isolated by thin layer chromatography. These compounds were found to be effective in inhibiting the growth of Gram-positive and Gram-negative bacteria but notCandida albicans. Their antimicrobial activity was similar to that of antibactericidal antibiotic streptomycin. They were found to inhibit the growth of the tested microbes at the minimum inhibition concentration of 0.09 mg/mL, and toxicity test using brine shrimp showed that even the low concentration of 0.09 mg/mL was very lethal towards the brine shrimps with 100% mortality rate. This study hence indicated thatin vitrocultured plantlets ofA. annuacan be used as the alternative method for production of artemisinin and its precursor with antimicrobial activities.

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The IMPDH inhibitor merimepodib provided in combination with the adenosine analogue remdesivir reduces SARS-CoV-2 replication to undetectable levels in vitro

F1000Research ◽

10.12688/f1000research.23639.1 ◽

2020 ◽

Vol 9 ◽

pp. 361

Author(s):

Natalya Bukreyeva ◽

Rachel A. Sattler ◽

Emily K. Mantlo ◽

Timothy Wanninger ◽

John T. Manning ◽

...

Keyword(s):

Disease Burden ◽

Vero Cells ◽

The United States ◽

The Novel ◽

Infectious Titer ◽

Adenosine Analogue ◽

Pre Treatment ◽

Novel Coronavirus ◽

Dose Dependent

Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) is the novel coronavirus responsible for the ongoing COVID-19 pandemic, which has resulted in over 2.5 million confirmed cases and 170,000 deaths worldwide as of late April 2020. The pandemic currently presents major public health and economic burdens worldwide. No vaccines or therapeutics have been approved for use to treat COVID-19 cases in the United States despite the growing disease burden, thus creating an urgent need for effective treatments. The adenosine analogue remdesivir (REM) has recently been investigated as a potential treatment option, and has shown some activity in limiting SARS-CoV-2 replication. We previously reported that the IMPDH inhibitor merimepodib (MMPD) provides a dose-dependent suppression of SARS-CoV-2 replication in vitro. Here, we report that a 4-hour pre-treatment of Vero cells with 2.5µM MMPD reduces the infectious titer of SARS-CoV-2 more effectively than REM at the same concentration. Additionally, pre-treatment of Vero cells with both REM and MMPD in combination reduces the infectious titer of SARS-CoV-2 to values below the detectable limit of our TCID50 assay. This result was achieved with concentrations as small as 1.25 µM MMPD and 2.5 µM REM. At concentrations of each agent as low as 0.31 µM, significant reduction of viral production occurred. This study provides evidence that REM and MMPD administered in combination might be an effective treatment for COVID-19 cases.

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1357. A Combination of Itraconazole and Amiodarone Is Highly Effective Against Trypanosoma cruzi Infection of Human Stem Cell-Derived Cardiomyocytes

Open Forum Infectious Diseases ◽

10.1093/ofid/ofy210.1188 ◽

2018 ◽

Vol 5 (suppl_1) ◽

pp. S415-S416

Author(s):

Gabriele Sass ◽

Roy Madigan ◽

Adriana Bozzi ◽

Nazish Sayed ◽

Joseph Wu ◽

...

Keyword(s):

Trypanosoma Cruzi ◽

Chagas Disease ◽

Cell Metabolism ◽

Vero Cells ◽

The United States ◽

Etiologic Agent ◽

Sterol Synthesis ◽

Xtt Assay ◽

Cruzi Infection

Abstract Background Trypanosoma cruzi is the etiologic agent of Chagas disease, which can result in severe cardiomyopathy. Trypanosoma cruzi is endemic to the Americas, and of particular importance in Latin America. In the United States and other nonendemic countries, rising case numbers have been observed. The only drugs available so far are benznidazole and nifurtimox, which have limited efficacy during chronic infection. We repurposed itraconazole, originally an antifungal, in combination with amiodarone, an antiarrhythmic, with the goal to interfere with Tc infection. Both drugs inhibit sterol synthesis, while amiodarone also inhibits calcium metabolism of Trypanosoma cruzi. Methods Human pluripotent stem cells (HiPSC) were differentiated to cardiomyocytes (HiPSC-CM). Vero cells or HiPSC-CM were infected with the T. cruzi trypomastigotes Y strain in the presence of itraconazole and/or amiodarone. After 48 hours, infection and multiplication were evaluated by Giemsa stain. Benznidazole was used as a reference compound. Cell viability was verified by XTT assay. Results Itraconazole and amiodarone showed dose-dependent interference with T. cruzi infection of Vero cells or HiPSC-CM. The combination of itraconazole and amiodarone was more potent than the single substances, or benznidazole at therapeutic concentrations, without affecting host cell metabolism. In addition to effects on infection, itraconazole, or amiodarone affected T. cruzi multiplication. Here, itraconazole/amiodarone combinations were more potent than either alone, both, in Vero cells, and HiPSC-CM. Conclusion Our in vitro data suggest that a combination of itraconazole and amiodarone might serve as an effective new treatment option for Chagas disease, particularly cardiac involvement, in human and animal patients. Disclosures All authors: No reported disclosures.

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Identification of Novel Bioactive Compound Derived from Rheum officinalis against Campylobacter jejuni NCTC11168

The Scientific World JOURNAL ◽

10.1155/2020/3591276 ◽

2020 ◽

Vol 2020 ◽

pp. 1-9

Author(s):

Mohammed Yosri ◽

Basma H. Amin ◽

Nermine N. Abed ◽

Amal S. Elithy ◽

Sayed M. Kareem ◽

...

Keyword(s):

Campylobacter Jejuni ◽

Bacterial Resistance ◽

Higher Plants ◽

Human Lymphocytes ◽

Bioactive Compound ◽

Vero Cells ◽

Infrared Analysis ◽

World Population ◽

Gastric Diseases

Gastric diseases are increasing with the infection of Campylobacter jejuni. Late stages of infection lead to peptic ulcer and gastric carcinoma. C. jejuni infects people within different stages of their life, especially childhood, causing severe diarrhea; it infects around two-thirds of the world population. Due to bacterial resistance against standard antibiotic, a new strategy is needed to impede Campylobacter infections. Plants provide highly varied structures with antimicrobial use which are unlikely to be synthesized in laboratories. A special feature of higher plants is their ability to produce a great number of organic chemicals of high structural diversity, the so-called secondary metabolites. Twenty plants were screened to detect their antibacterial activities. Screening results showed that Rheum officinalis was the most efficient against C. jejuni. Fractionation pattern was obtained by column chromatography, while the purity test was done by thin-layer chromatography (TLC). The chemical composition of bioactive compound was characterized using GC-MS, nuclear magnetic resonance, and infrared analysis. Minimal inhibitory concentration (MIC) of the purified compound was 31.25 µg/ml. Cytotoxicity assay on Vero cells was evaluated to be 497 µg/ml. Furthermore, the purified bioactive compound activated human lymphocytes in vitro. The data presented here show that Rheum officinalis could potentially be used in modern applications aimed at the treatment or prevention of foodborne diseases.

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The Ɛ34 Phage Tailspike Protein: An in vitro characterization, Structure Prediction, Potential Interaction with S. newington LPS and Cytotoxicity Assessment to Animal Cell Line.

10.1101/2021.09.20.461090 ◽

2021 ◽

Author(s):

Joseph A Ayariga ◽

Logan Gildea ◽

Honghzua Wu ◽

Robert Villafane

Keyword(s):

Structure Prediction ◽

Animal Cell ◽

Specific Interaction ◽

Vero Cells ◽

Potential Interaction ◽

O Antigen ◽

In Vitro Characterization ◽

Cytotoxicity Assessment ◽

Tailspike Protein

The E34 phage is a member of the podoviridae family of phages, (short non-contractile tailed bacteriophages) that uses Salmonella newington as its host. This phage initiates the infection of its host via a specific interaction between its tailspike protein (TSP) and the lipopolysaccharides (LPS) of the bacterial. The E34 TSP is structurally similar and functionally equivalent to the P22 phage whose TSP has been well characterized and electron micrographs of both phages appear indistinguishable. The crystal structure of P22 phage TSP in complex with the O-antigen of S. typhimurium has been determined; and the active site of the TSP demonstrated to be the residues Asp392, Asp395 and Glu359 of the receptor binding domain. In another phage called E15, a phylogenetic relative of E34 phage, a short polysaccharide consisting of alpha-Gal-Man-Rha repeating units is responsible for the interaction between the E15 phage and Salmonella anatum LPS leading to the adsorption of the phage to the bacteria. Studies on E34 phage shows that it interacts with Salmonella newington O antigen polysaccharide component of the LPS, this polysaccharide consists of mannosyl-rhamnosyl-galactose repeating units joined together by beta-galactosyl linkages. However, no data exist regarding the specific residues of E34 TSP that are responsible for LPS binding and hydrolysis. In this study, the tailspike gene was cloned onto vector pET30a-LIC and expressed as a fusion protein termed the extended E34 TSP (EE34 TSP). We characterized the protein based on resistance to heat, SDS, and proteases; showing that the protein is heat resistant, shows aberrant electrophoretic mobility in the presence of SDS gradient, and actively binds to P22 phage heads to form hybrid phages that cannot infect P22 host. We also demonstrate via in silico study that the E34 TSP binds to and hydrolyses the O-antigen of its host via the ALA250, SER279 and ASP280 residues. Finally, testing E34 phage ability to protect Vero cells from Salmonella infection shows highly encouraging results, implying that E34 phage can be used in therapeutic or preventive medicine.

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Curcumin Suppresses Epithelial Growth Factor Receptor (EGFR) and Proliferative Protein (Ki 67) in Acute Lung Injury and Lung Fibrosis In vitro and In vivo

Endocrine Metabolic & Immune Disorders - Drug Targets ◽

10.2174/1871530319666190823160230 ◽

2020 ◽

Vol 20 (4) ◽

pp. 558-563 ◽

Cited By ~ 4

Author(s):

Sadiya Bi Shaikh ◽

Ashwini Prabhu ◽

Yashodhar Prabhakar Bhandary

Keyword(s):

Acute Lung Injury ◽

Pulmonary Fibrosis ◽

Lung Injury ◽

Growth Factor Receptor ◽

Bioactive Compound ◽

The Elderly ◽

Alveolar Epithelial Cells ◽

Ki 67

Background:: Acute lung injury is one of the common conditions caused due to bleomycin therapy which leads to pulmonary fibrosis, which is one of the severe interstitial lung diseases most commonly affecting the elderly individuals. EGFR and Ki67 can be marked as beneficial markers for detecting pulmonary fibrosis based on which clinicians can guide the therapy. Objective:: The aim of the study was to evaluate the effect of curcumin as an intervention on two prognostic markers EGFR and Ki67 in bleomycin-induced basal alveolar epithelial cells and C57BL/6 mice. Protein expressions and pathological expressions of EGFR and Ki67 were evaluated to analyze the effect of curcumin via both in vitro and in vivo approaches. Methods:: The effect of curcumin was investigated both on cell lines (A549) and animal model (both normal and bleomycin-induced mice, n=6) via techniques like western blotting for protein expression. Techniques like immunofluorescence and immunohistochemistry were carried out and examined under confocal microscopy and phase contrast microscopy to analyze the expressions of the said biomarkers. Bleomycin was used as a causative agent to induce inflammation. Results:: The natural polyphenol curcumin could downregulate the expressions levels of Ki67 and EGFR both in vitro and in vivo. Immunofluorescence analysis of proliferative marker Ki67 showed a reduced expression on curcumin treatment in vitro. The pathological sections from treated lungs showed a significant decrease in EGFR and Ki67 levels when exposed to curcumin. Conclusion:: We conclude that curcumin, a well-known natural bioactive compound holds strong antiproliferative effects on Ki67 and EGFR expressions.We observed that a clinical outcome in the diagnosis of pulmonary fibrosis remains to be unconvincing so far. Curcumin can be considered as a potential therapeutic.

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Correlative microscopy in distrubuted (client/server) computing environments: tools for catalyzing the rapid dissemination of information about the cell biology of the human prostate

Proceedings, annual meeting, Electron Microscopy Society of America ◽

10.1017/s0424820100139780 ◽

1995 ◽

Vol 53 ◽

pp. 682-683

Author(s):

A. Hakam ◽

J.T. Gau ◽

M.L. Grove ◽

B.A. Evans ◽

M. Shuman ◽

...

Keyword(s):

United States ◽

Cell Biology ◽

Tissue Bank ◽

The United States ◽

Prostate Adenocarcinoma ◽

Correlative Microscopy ◽

Client Server ◽

Critical Elements ◽

Transplant Organ

Prostate adenocarcinoma is the most common malignant tumor of men in the United States and is the third leading cause of death in men. Despite attempts at early detection, there will be 244,000 new cases and 44,000 deaths from the disease in the United States in 1995. Therapeutic progress against this disease is hindered by an incomplete understanding of prostate epithelial cell biology, the availability of human tissues for in vitro experimentation, slow dissemination of information between prostate cancer research teams and the increasing pressure to “ stretch” research dollars at the same time staff reductions are occurring.To meet these challenges, we have used the correlative microscopy (CM) and client/server (C/S) computing to increase productivity while decreasing costs. Critical elements of our program are as follows:1) Establishing the Western Pennsylvania Genitourinary (GU) Tissue Bank which includes >100 prostates from patients with prostate adenocarcinoma as well as >20 normal prostates from transplant organ donors.

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The Unusual Cruelty of Nursing Homes Behind Bars

Federal Sentencing Reporter ◽

10.1525/fsr.2020.32.5.264 ◽

2020 ◽

Vol 32 (5) ◽

pp. 264-271

Author(s):

Rachel E. López

Keyword(s):

Nursing Homes ◽

Human Dignity ◽

Prison Population ◽

Financial Burden ◽

The Elderly ◽

The United States ◽

Term Care ◽

Trained Staff ◽

State Prisons ◽

The Cost

The elderly prison population continues to rise along with higher rates of dementia behind bars. To maintain the detention of this elderly population, federal and state prisons are creating long-term care units, which in turn carry a heavy financial burden. Prisons are thus gearing up to become nursing homes, but without the proper trained staff and adequate financial support. The costs both to taxpayers and to human dignity are only now becoming clear. This article squarely addresses the second dimension of this carceral practice, that is the cost to human dignity. Namely, it sets out why indefinitely incarcerating someone with dementia or other neurocognitive disorders violates the Eighth Amendment of the United States Constitution’s prohibition on cruel and unusual punishment. This conclusion derives from the confluence of two lines of U.S. Supreme Court precedent. First, in Madison v. Alabama, the Court recently held that executing someone (in Madison’s case someone with dementia) who cannot rationally understand their sentence amounts to cruel and unusual punishment. Second, in line with Miller v. Alabama, which puts life without parole (LWOP) sentences in the same class as death sentences due to their irrevocability, this holding should be extended to LWOP sentences. Put another way, this article explains why being condemned to life is equivalent to death for someone whose neurodegenerative disease is so severe that they cannot rationally understand their punishment.

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