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Fulltext Combating Acanthamoeba spp. cysts: what are the options?

Anwar A, Khan NA, Siddiqui R

Parasit Vectors, 2018 01 09;11(1):26.
PMID: 29316961 DOI: 10.1186/s13071-017-2572-z

Acanthamoeba spp. are protist pathogens and causative agents of serious infections including keratitis and granulomatous amoebic encephalitis. Its ability to convert into dormant and highly resistant cysts form limits effectiveness of available therapeutic agents and presents a pivotal challenge for drug development. During the cyst stage, Acanthamoeba is protected by the presence of hardy cyst walls, comprised primarily of carbohydrates and cyst-specific proteins, hence synthesis inhibition and/or degradation of cyst walls is of major interest. This review focuses on targeting of Acanthamoeba cysts by identifying viable therapeutic targets.

Matched MeSH terms: Amebiasis/epidemiology*; Amebiasis/prevention & control*
Fulltext Cobalt nanoparticles as novel nanotherapeutics against Acanthamoeba castellanii

Anwar A, Numan A, Siddiqui R, Khalid M, Khan NA

Parasit Vectors, 2019 Jun 03;12(1):280.
PMID: 31159839 DOI: 10.1186/s13071-019-3528-2

BACKGROUND: Species of Acanthamoeba are facultative pathogens which can cause sight threatening Acanthamoeba keratitis and a rare but deadly brain infection, granulomatous amoebic encephalitis. Due to conversion of Acanthamoeba trophozoites to resistant cyst stage, most drugs are found to be ineffective at preventing recurrence of infection. This study was designed to test the antiacanthamoebic effects of different cobalt nanoparticles (CoNPs) against trophozoites and cysts, as well as parasite-mediated host cell cytotoxicity.
METHODS: Three different varieties of CoNPs were synthesized by utilizing hydrothermal and ultrasonication methods and were thoroughly characterized by X-ray diffraction and field emission scanning electron microscopy. Amoebicidal, encystation, excystation, and host cell cytopathogenicity assays were conducted to study the antiacanthamoebic effects of CoNPs.
RESULTS: The results of the antimicrobial evaluation revealed that cobalt phosphate Co3(PO4)2 hexagonal microflakes, and 100 nm large cobalt hydroxide (Co(OH)2) nanoflakes showed potent amoebicidal activity at 100 and 10 µg/ml against Acanthamoeba castellanii as compared to granular cobalt oxide (Co3O4) of size 35-40 nm. Furthermore, encystation and excystation assays also showed consistent inhibition at 100 µg/ml. CoNPs also inhibited amoebae-mediated host cell cytotoxicity as determined by lactate dehydrogenase release without causing significant damage to human cells when treated alone.
CONCLUSIONS: To our knowledge, these findings determined, for the first time, the effects of composition, size and morphology of CoNPs against A. castellanii. Co3(PO4)2 hexagonal microflakes showed the most promising antiamoebic effects as compared to Co(OH)2 nanoflakes and granular Co3O4. The results reported in the present study hold potential for the development of antiamoebic nanomedicine.

Matched MeSH terms: Amebiasis/drug therapy
Fulltext Cecal amebiasis mimicking inflammatory bowel disease

Cheng CW, Feng CM, Chua CS

J Int Med Res, 2020 May;48(5):300060520922379.
PMID: 32475192 DOI: 10.1177/0300060520922379

Amebiasis is a frequently occurring parasitic infection in South East Asia. We present a case of a 54-year-old man with right lower quadrant abdominal pain that persisted for longer than 1 year. He had been diagnosed with inflammatory bowel disease in Indonesia. His abdominal pain persisted, despite therapy, and he visited Malaysia for transnational medical advice. Abdominal ultrasound showed fatty liver, gallbladder polyps, and a small left renal stone. Colonoscopy showed multiple ulcers in the cecum and a histopathological examination confirmed amebic infection of the cecum. The colonic ulcers subsided after anti-amebic treatment. This case highlights the need to consider the differential diagnosis of amebic colitis in patients presenting with manifestations of inflammatory bowel disease, especially in patients who live in or have traveled to endemic areas.

Matched MeSH terms: Amebiasis/diagnosis*; Amebiasis/physiopathology
Fulltext Brine shrimp lethality and acute oral toxicity studies on Swietenia mahagoni (Linn.) Jacq. seed methanolic extract

Sahgal G, Ramanathan S, Sasidharan S, Mordi MN, Ismail S, Mansor SM

Pharmacognosy Res, 2010 Jul;2(4):215-20.
PMID: 21808570 DOI: 10.4103/0974-8490.69107

The seeds of Swietenia mahagoni have been applied in folk medicine for the treatment of hypertension, diabetes, malaria, amoebiasis, cough, chest pain, and intestinal parasitism. Here we are the first to report on the toxicity of the Swietenia mahagoni crude methanolic (SMCM) seed extract.

Matched MeSH terms: Amebiasis
Brain-eating Amoebae Infection: Challenges and Opportunities in Chemotherapy

Mungroo MR, Anwar A, Khan NA, Siddiqui R

Mini Rev Med Chem, 2019;19(12):980-987.
PMID: 30868950 DOI: 10.2174/1389557519666190313161854

Pathogenic free-living amoeba are known to cause a devastating infection of the central nervous system and are often referred to as "brain-eating amoebae". The mortality rate of more than 90% and free-living nature of these amoebae is a cause for concern. It is distressing that the mortality rate has remained the same over the past few decades, highlighting the lack of interest by the pharmaceutical industry. With the threat of global warming and increased outdoor activities of public, there is a need for renewed interest in identifying potential anti-amoebic compounds for successful prognosis. Here, we discuss the available chemotherapeutic options and opportunities for potential strategies in the treatment and diagnosis of these life-threatening infections.

Matched MeSH terms: Amebiasis/diagnosis; Amebiasis/drug therapy*; Amebiasis/parasitology*
Fulltext Brain-Eating Amoebae: Predilection Sites in the Brain and Disease Outcome

Ong TYY, Khan NA, Siddiqui R

J Clin Microbiol, 2017 07;55(7):1989-1997.
PMID: 28404683 DOI: 10.1128/JCM.02300-16

Acanthamoeba spp. and Balamuthia mandrillaris are causative agents of granulomatous amoebic encephalitis (GAE), while Naegleria fowleri causes primary amoebic meningoencephalitis (PAM). PAM is an acute infection that lasts a few days, while GAE is a chronic to subacute infection that can last up to several months. Here, we present a literature review of 86 case reports from 1968 to 2016, in order to explore the affinity of these amoebae for particular sites of the brain, diagnostic modalities, treatment options, and disease outcomes in a comparative manner.

Matched MeSH terms: Amebiasis/pathology*
Aryl Quinazolinone Derivatives as Novel Therapeutic Agents against Brain-Eating Amoebae

Mungroo MR, Shahbaz MS, Anwar A, Saad SM, Khan KM, Khan NA, et al.

ACS Chem Neurosci, 2020 08 19;11(16):2438-2449.
PMID: 31961126 DOI: 10.1021/acschemneuro.9b00596

Naegleria fowleri and Balamuthia mandrillaris are protist pathogens that infect the central nervous system, causing primary amoebic meningoencephalitis and granulomatous amoebic encephalitis with mortality rates of over 95%. Quinazolinones and their derivatives possess a wide spectrum of biological properties, but their antiamoebic effects against brain-eating amoebae have never been tested before. In this study, we synthesized a variety of 34 novel arylquinazolinones derivatives (Q1-Q34) by altering both quinazolinone core and aryl substituents. To study the antiamoebic activity of these synthetic arylquinazolinones, amoebicidal and amoebistatic assays were performed against N. fowleri and B. mandrillaris. Moreover, amoebae-mediated host cells cytotopathogenicity and cytotoxicity assays were performed against human keratinocytes cells in vitro. The results revealed that selected arylquinazolinones derivatives decreased the viability of B. mandrillaris and N. fowleri significantly (P < 0.05) and reduced cytopathogenicity of both parasites. Furthermore, these compounds were also found to be least cytotoxic against HaCat cells. Considering that nanoparticle-based materials possess potent in vitro activity against brain-eating amoebae, we conjugated quinazolinones derivatives with silver nanoparticles and showed that activities of the drugs were enhanced successfully after conjugation. The current study suggests that quinazolinones alone as well as conjugated with silver nanoparticles may serve as potent therapeutics against brain-eating amoebae.

Matched MeSH terms: Amebiasis
Fulltext Antimicrobial activities of green synthesized gums-stabilized nanoparticles loaded with flavonoids

Anwar A, Masri A, Rao K, Rajendran K, Khan NA, Shah MR, et al.

Sci Rep, 2019 02 28;9(1):3122.
PMID: 30816269 DOI: 10.1038/s41598-019-39528-0

Herein, we report green synthesized nanoparticles based on stabilization by plant gums, loaded with citrus fruits flavonoids Hesperidin (HDN) and Naringin (NRG) as novel antimicrobial agents against brain-eating amoebae and multi-drug resistant bacteria. Nanoparticles were thoroughly characterized by using zetasizer, zeta potential, atomic force microscopy, ultravoilet-visible and Fourier transform-infrared spectroscopic techniques. The size of these spherical nanoparticles was found to be in the range of 100-225 nm. The antiamoebic effects of these green synthesized Silver and Gold nanoparticles loaded with HDN and NRG were tested against Acanthamoeba castellanii and Naegleria fowleri, while antibacterial effects were evaluated against methicillin-resistant Staphylococcus aureus (MRSA) and neuropathogenic Escherichia coli K1. Amoebicidal assays revealed that HDN loaded Silver nanoparticles stabilized by gum acacia (GA-AgNPs-HDN) quantitatively abolished amoeba viability by 100%, while NRG loaded Gold nanoparticles stabilized by gum tragacanth (GT-AuNPs-NRG) significantly reduced the viability of A. castellanii and N. fowleri at 50 µg per mL. Furthermore, these nanoparticles inhibited the encystation and excystation by more than 85%, as well as GA-AgNPs-HDN only completely obliterated amoeba-mediated host cells cytopathogenicity. Whereas, GA-AgNPs-HDN exhibited significant bactericidal effects against MRSA and E. coli K1 and reduced bacterial-mediated host cells cytotoxicity. Notably, when tested against human cells, these nanoparticles showed minimal (23%) cytotoxicity at even higher concentration of 100 µg per mL as compared to 50 µg per mL used for antimicrobial assays. Hence, these novel nanoparticles formulations hold potential as therapeutic agents against infections caused by brain-eating amoebae, as well as multi-drug resistant bacteria, and recommend a step forward in drug development.

Matched MeSH terms: Amebiasis/drug therapy
Fulltext Antiamoebic activity of plant-based natural products and their conjugated silver nanoparticles against Acanthamoeba castellanii (ATCC 50492)

Anwar A, Ting ELS, Anwar A, Ain NU, Faizi S, Shah MR, et al.

AMB Express, 2020 Feb 03;10(1):24.
PMID: 32016777 DOI: 10.1186/s13568-020-0960-9

Acanthamoeba spp. are the causative agent of Acanthamoeba keratitis and granulomatous amoebic encephalitis (GAE). The current options to treat Acanthamoeba infections have limited success. Silver nanoparticles show antimicrobial effects and enhance the efficacy of their payload at the specific biological targets. Natural folk plants have been widely used for treating diseases as the phytochemicals from several plants have been shown to exhibit amoebicidal effects. Herein, we used natural products of plant or commercial sources including quercetin (QT), kolavenic acid (PGEA) isolated from plant extracts of Polyalthia longifolia var pendula and crude plant methanolic extract of Caesalpinia pulcherrima (CPFLM) as antiacanthamoebic agents. Furthermore, these plant-based materials were conjugated with silver nanoparticles (AgNPs) to determine the effects of the natural compounds and their nanoconjugates against a clinical isolate of A. castellanii from a keratitis patient (ATCC 50492) belonging to the T4 genotype. The compounds were conjugated with AgNPs and characterized by using ultraviolet visible spectrophotometry and atomic force microscopy. Quercetin coated silver nanoparticles (QT-AgNPs) showed characteristic surface plasmon resonance band at 443 nm and the average size distribution was found to be around 45 nm. The natural compounds alone and their nanoconjugates were tested for the viability of amoebae, encystation and excystation activity against A. castellanii. The natural compounds showed significant growth inhibition of A. castellanii while QT-AgNPs specifically exhibited enhanced antiamoebic effects as well as interrupted the encystation and excystation activity of the amoebae. Interestingly, these compounds and nanoconjugates did not exhibit in vitro cytotoxic effects against human cells. Plant-based compounds and extracts could be an interesting strategy in development of alternative therapeutics against Acanthamoeba infections.

Matched MeSH terms: Amebiasis
Antiamoebic activity of 3-aryl-6,7-dimethoxyquinazolin-4(3H)-one library against Acanthamoeba castellanii

Shahbaz MS, Anwar A, Saad SM, Kanwal, Anwar A, Khan KM, et al.

Parasitol Res, 2020 Jul;119(7):2327-2335.
PMID: 32476058 DOI: 10.1007/s00436-020-06710-7

Acanthamoeba castellanii is a free-living amoeba which can cause a blinding keratitis and fatal granulomatous amoebic encephalitis. The treatment of Acanthamoeba infections is challenging due to formation of cyst. Quinazolinones are medicinally important scaffold against parasitic diseases. A library of nineteen new 3-aryl-6,7-dimethoxyquinazolin-4(3H)-one derivatives was synthesized to evaluate their antiamoebic activity against Acanthamoeba castellanii. One-pot synthesis of 3-aryl-6,7-dimethoxyquinazolin-4(3H)-ones (1-19) was achieved by reaction of 2-amino-4,5-dimethoxybenzoic acid, trimethoxymethane, and different substituted anilines. These compounds were purified and characterized by standard chromatographic and spectroscopic techniques. Antiacanthamoebic activity of these compounds was determined by amoebicidal, encystation, excystation and host cell cytopathogenicity in vitro assays at concentrations of 50 and 100 μg/mL. The IC50 was found to be between 100 and 50 μg/mL for all the compounds except compound 5 which did not exhibit amoebicidal effects at these concentrations. Furthermore, lactate dehydrogenase assay was also performed to evaluate the in vitro cytotoxicity of these compounds against human keratinocyte (HaCaT) cells. The results revealed that eighteen out of nineteen derivatives of quinazolinones significantly decreased the viability of A. castellanii. Furthermore, eighteen out of nineteen tested compounds inhibited the encystation and excystation, as well as significantly reduced the A. castellanii-mediated cytopathogenicity against human cells. Interestingly, while tested against human normal cell line HaCaT keratinocytes, all compounds did not exhibit any overt cytotoxicity. Furthermore, a detailed structure-activity relationship is also studied to optimize the most potent hit from these synthetic compounds. This report presents several potential lead compounds belonging to 3-aryl-6,7-dimethoxyquinazolin-4(3H)-one derivatives for drug discovery against infections caused by Acanthamoeba castellanii.

Matched MeSH terms: Amebiasis/drug therapy; Amebiasis/parasitology
Fulltext Amoebic colitis with liver abscess

Mansharan Kaur Chaincel Singh

International e-Journal of Science, Medicine & Education, 2018;12(1):27-31.
MyJurnal

Amoebiasis is a parasitic infection caused by the
intestinal protozoan Entamoeba histolytica, most
prevalent in developing countries. It results in 40,000 to
100,000 deaths each year from amoebic colitis and extra
intestinal infections. Amoebic liver abscess (ALA)
is the most common extra intestinal site of infection
with an incidence of between 3% and 9% of all cases of
amoebiasis. Ultrasound which has a sensitivity of more
than 90% for detecting ALA is highly recommended
as an initial investigation followed by serological
demonstration of circulating antibodies specific to
Entamoeba histolytica.

Matched MeSH terms: Amebiasis
Fulltext Amoebiasis: a 10 year retrospective study at the University Hospital, Kuala Lumpur

Jamaiah I, Shekhar KC

Med J Malaysia, 1999 Sep;54(3):296-302.
PMID: 11045054

A hospital based retrospective study of amoebiasis was carried out for a ten-year period at the University Hospital, Kuala Lumpur. Of the 51 cases traced, 30 (59%) had amoebic dysentery, 20 (39%) were amoebic liver abscess (ALA) and one patient had both conditions. Entameoba histolytica trophozoites were identified in 13 (43%) of the amoebic dysenteric stools and 9 (30%) from biopsy. Of the 20 (39%) ALA cases, only one showed parasites in the stool and biopsy. Majority of the patients with dysentery were Malays while Chinese comprised 40% with ALA. Males predominated overall with a male female ratio of 3:1, while for ALA it was 9:1. Most of ALA were single (71.4%) and were localised in the right lobe. The majority of the patients were unemployed. Eighty three percent (83%) of the patients presented with diarrhoea or dysentery followed by abdominal pain while those with ALA had fever, chills, rigors and pain in the right hypochondrium. Eighty percent of the ALA cases showed hepatomegaly. All patients responded to treatment with metronidazole.

Matched MeSH terms: Amebiasis/epidemiology*
Fulltext Acanthamoeba in Southeast Asia - Overview and Challenges

Bunsuwansakul C, Mahboob T, Hounkong K, Laohaprapanon S, Chitapornpan S, Jawjit S, et al.

Korean J Parasitol, 2019 Aug;57(4):341-357.
PMID: 31533401 DOI: 10.3347/kjp.2019.57.4.341

Acanthamoeba, one of free-living amoebae (FLA), remains a high risk of direct contact with this protozoan parasite which is ubiquitous in nature and man-made environment. This pathogenic FLA can cause sight-threatening amoebic keratitis (AK) and fatal granulomatous amoebic encephalitis (GAE) though these cases may not commonly be reported in our clinical settings. Acanthamoeba has been detected from different environmental sources namely; soil, water, hot-spring, swimming pool, air-conditioner, or contact lens storage cases. The identification of Acanthamoeba is based on morphological appearance and molecular techniques using PCR and DNA sequencing for clinico-epidemiological purposes. Recent treatments have long been ineffective against Acanthamoeba cyst, novel anti-Acanthamoeba agents have therefore been extensively investigated. There are efforts to utilize synthetic chemicals, lead compounds from medicinal plant extracts, and animal products to combat Acanthamoeba infection. Applied nanotechnology, an advanced technology, has shown to enhance the anti-Acanthamoeba activity in the encapsulated nanoparticles leading to new therapeutic options. This review attempts to provide an overview of the available data and studies on the occurrence of pathogenic Acanthamoeba among the Association of Southeast Asian Nations (ASEAN) members with the aim of identifying some potential contributing factors such as distribution, demographic profile of the patients, possible source of the parasite, mode of transmission and treatment. Further, this review attempts to provide future direction for prevention and control of the Acanthamoeba infection.

Matched MeSH terms: Amebiasis/diagnosis; Amebiasis/epidemiology*; Amebiasis/therapy; Amebiasis/transmission
A proposed cascade of vascular events leading to granulomatous amoebic encephalitis

Baig AM, Khan NA

Microb Pathog, 2015 Nov;88:48-51.
PMID: 26276705 DOI: 10.1016/j.micpath.2015.08.005

Granulomatous amoebic encephalitis due to Acanthamoeba is a chronic disease that almost always results in death. Hematogenous spread is a pre-requisite followed by amoebae invasion of the blood-brain barrier to enter the central nervous system. Given the systemic nature of this infection, a significant latent period of several months before the appearance of clinical manifestations is puzzling. Based on reported cases, here we propose pathogenetic mechanisms that explain the above described latency of the disease.

Matched MeSH terms: Amebiasis
A colonic amoebic abscess mimicking colonic carcinoma

Tan LP, Foong KK, Yvonne Ai LL

Med J Malaysia, 2018 10;73(5):334-335.
PMID: 30350818 MyJurnal

Amebiasis is one of the major causes of diarrhea in the developing countries and it can present with a wide range of gastrointestinal symptoms depending on the phase of infection. We described a case of 50 year-old male patient who presented with abdominal pain, diarrhea and vomiting. After right hemicolectomy for appendicular abscess with tumour over the ileum, histopathological examinations revealed numerous trophozoites of Entamoeba histolytica in a background of inflammations (Figure 1). Following resection of the ameboma, he received intravenous metronidazole treatment for total of two weeks duration.

Matched MeSH terms: Amebiasis

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