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Sample records for acanthamoeba astigmatism blepharitis

  1. Blepharitis

    MedlinePlus

    ... this page: //medlineplus.gov/ency/article/001619.htm Blepharitis To use the sharing features on this page, please enable JavaScript. Blepharitis is inflamed, irritated, itchy, and reddened eyelids. It ...

  2. Blepharitis.

    PubMed

    Bernardes, Taliana Freitas; Bonfioli, Adriana Alvim

    2010-05-01

    Blepharitis is a chronic inflammatory process of the eyelid margin. It is a common eye disorder throughout the world and can affect any age group. It may be associated with several systemic diseases, particularly rosacea and seborrheic dermatitis, and is related to other ocular conditions like dry eye, chalazion, conjunctivitis, and keratitis. Common symptoms associated with blepharitis are burning sensation, irritation, tearing, photophobia, blurred vision, and red eyes. Clinical examination reveals the presence of scurf, telangiectatic vascular changes of the eyelid margin, inspissated meibomian glands, conjuntival hyperemia, punctuate keratopathy, cornea vascularization, and ulceration. Patients with longstanding chronic blepharitis may present hypertrophy of the lid margin, scars, madarosis, trichiasis, and poliosis. Treatment of blepharitis is long and unsatisfactory. Long-term commitment to eyelid hygiene is essential. Other treatment options are discussed.

  3. Interventions for chronic blepharitis

    PubMed Central

    Lindsley, Kristina; Matsumura, Sueko; Hatef, Elham; Akpek, Esen K

    2012-01-01

    Background Blepharitis, an inflammatory condition associated with itchiness, redness, flaking, and crusting of the eyelids, is a common eye condition that affects both children and adults. It is common in all ethnic groups and across all ages. Although infrequent, blepharitis can lead to permanent alterations to the eyelid margin or vision loss from superficial keratopathy (abnormality of the cornea), corneal neovascularization, and ulceration. Most importantly, blepharitis frequently causes significant ocular symptoms such as burning sensation, irritation, tearing, and red eyes as well as visual problems such as photophobia and blurred vision. The exact etiopathogenesis is unknown, but suspected to be multifactorial, including chronic low-grade infections of the ocular surface with bacteria, infestations with certain parasites such as demodex, and inflammatory skin conditions such as atopy and seborrhea. Blepharitis can be categorized in several different ways. First, categorization is based on the length of disease process: acute or chronic blepharitis. Second, categorization is based on the anatomical location of disease: anterior, or front of the eye (e.g. staphylococcal and seborrheic blepharitis), and posterior, or back of the eye (e.g. meibomian gland dysfunction (MGD)). This review focuses on chronic blepharitis and stratifies anterior and posterior blepharitis. Objectives To examine the effectiveness of interventions in the treatment of chronic blepharitis. Search methods We searched CENTRAL (which contains the Cochrane Eyes and Vision Group Trials Register) (The Cochrane Library 2012, Issue 1), MEDLINE (January 1950 to February 2012), EMBASE (January 1980 to February 2012), the metaRegister of Controlled Trials (mRCT) (www.controlled-trials.com), ClinicalTrials.gov (www.clinicaltrials.gov) and the WHO International Clinical Trials Registry Platform (ICTRP) (www.who.int/ictrp/search/en). We searched the reference lists of included studies for any

  4. [Calcinosis conjunctivae in chronic blepharitis].

    PubMed

    Auw-Hädrich, C; Reinhard, T

    2008-01-01

    We present a case of a 53-year-old female patient with chronic blepharitis and conjunctival overgrowth of the cornea. Histologically the conjunctival overgrowth contained foreign body granulomas around calcification focuses. This could have been promoted by the application of phosphate-containing eye drops and the presence of chemokines like TNF-alpha, which occur in chronic blepharitis. Thus, we recommend the application of phosphate-free eye drops in patients with chronic blepharitis.

  5. Blepharitis: Always Remember Demodex

    PubMed Central

    Bhandari, Vipul; Reddy, Jagadeesh K.

    2014-01-01

    Purpose: To determine the incidence and density of Demodex species on the eyelashes of subjects with normal eyelids, anterior blepharitis (AB), meibomian-gland dysfunction (MGD), and mixed blepharitis (MB). Materials and Methods: One hundred and fifty consecutive patients diagnosed with AB, MGD, and MB were recruited. An additional 50 individuals were recruited who were free of lid and margin disease to serve as a control group. All patients underwent a standard eye examination. Data on ocular symptomatology were gathered. Digital photography was performed of the lid margins. Lash sampling was performed by epilating the lashes and the lashes were checked for Demodex based on morphology using light microscopy. The total number of mites were tabulated for each eye. Comparison between the affected eyes and the control eyes was performed. Data were analyzed using the Chi-square test. Results: A total of 200 patients were included. Twenty percenth had AB, 40% had MGD, and 40% had MB. The incidence of Demodex infestation was 90% in cases of AB, 60% in MGD cases, and 90% in MB cases. The incidence of Demodex in control subjects was 18%. Conclusions: The incidence and density of Demodex infestation was highest in patients with AB and MB. Lid irritation and presence of cylindrical dandruff were indicative of a high-density infestation. These signs should alert the clinician to treat concomitant Demodex infestation. PMID:25371637

  6. Discoid lupus erythematosus presenting as unilateral blepharitis.

    PubMed

    Au, Leon

    2006-01-01

    A 39-year-old man presented with a 4-month history of unilateral blepharitis that did not respond to conventional treatment. Punch biopsy confirmed the diagnosis of discoid lupus erythematosus. Unilateral blepharitis as the only presenting sign of discoid lupus erythematosus is uncommon but should be considered in the differential diagnosis in patients with asymmetric blepharitis.

  7. Postoperative astigmatism.

    PubMed

    Swinger, C A

    1987-01-01

    With the numerous significant advances in surgical methodology--e.g., microinstrumentation, the operating microscope, the surgical keratometer, and intraocular lenses--that have been developed over the past two decades, both surgeons and patients have become increasingly aware of the final optic result of any surgical intervention. This is especially so since the development of refractive surgery, where good uncorrected vision is frequently the final arbiter of success. We have progressed to the stage where the optic manipulation of the cornea, whether intentional or otherwise, can be understood in terms of a number of variables. These include the preparation and closure of the surgical wound, the choice of suture material, and both intraoperative and postoperative manipulations. Where these have failed and postoperative astigmatism still occurs, a number of surgical procedures are available to reduce the astigmatic error to an acceptable level.

  8. Pathogenic role of Demodex mites in blepharitis

    PubMed Central

    Liu, Jingbo; Sheha, Hosam; Tseng, Scheffer C.G.

    2010-01-01

    Purpose of review To summarize the key literature and our research experience regarding Demodex infestation as a potential cause of ocular inflammatory diseases with a special emphasis on Demodex blepharitis. Recent findings Two distinct Demodex species have been confirmed as a cause of blepharitis: Demodex folliculorum can cause anterior blepharitis associated with disorders of eyelashes, and D. brevis can cause posterior blepharitis with meibomian gland dysfunction and keratoconjunctivitis. Tea tree oil treatments with either 50% lid scrubs or 5% lid massages are effective in eradicating mites and reducing ocular surface inflammation. Summary Demodex blepharitis is a common but overlooked external eye disease. The pathogenesis of Demodex blepharitis in eliciting ocular surface inflammation has been further clarified. The modified eyelash sampling and counting method makes it easier and more accurate to diagnose Demodex infestation. Tea tree oil shows promising potential to treat Demodex blepharitis by reducing Demodex counts with additional antibacterial, antifungal, and anti-inflammatory actions. PMID:20689407

  9. [Progress of study on the demodex blepharitis].

    PubMed

    Zhang, X Y; Sun, X G

    2016-04-11

    Demodex blepharitis is an infectious ocular disease which is common, but always to be neglected. There were a few related reports about this disease in China. The pathogenic mechanism of demodex blepharitis is not clear at present. Literatures held different ideas about its diagnosis, therapeutic methods and curative effect. This review will summarize the new development of demodex blepharitis in order to provide references for its basic and clinical researches.(Chin J Ophthalmol, 2016, 52: 315-320).

  10. Keratophakia--postoperative astigmatism.

    PubMed

    Swinger, C A; Troutman, R C; Forman, J S

    1987-01-01

    Forty-nine cases of primary keratophakia and 13 cases of secondary keratophakia were analyzed for postoperative astigmatism. For primary cases, the surgically induced astigmatism was 1.55 D, whereas for secondary cases it was 0.19 D (insignificant). There was a tendency for both procedures to induce against-the-rule astigmatism, and both procedures were found capable of producing irregular astigmatism.

  11. Effects of Induced Astigmatism.

    ERIC Educational Resources Information Center

    Schubert, Delwyn G.; Walton, Howard N.

    1968-01-01

    The relationship of astigmatism to reading and the possible detrimental effects it might have on reading were investigated. The greatest incidence of astigmatism was for the with-the-rule type ranging from .50 to 1.00 diopter. This type of astigmatism was induced in 35 seniors from the Los Angeles College of Optometry by placing cylindrical lenses…

  12. Treatment of blepharitis: recent clinical trials.

    PubMed

    Pflugfelder, Stephen C; Karpecki, Paul M; Perez, Victor L

    2014-10-01

    Blepharitis is a chronic inflammatory disease of the eyelids that is frequently encountered in clinical practice. The etiology of the disorder is complex and not fully understood, but the general consensus is that bacteria and inflammation contribute to the pathology. Blepharitis can be classified into anterior blepharitis, involving the anterior lid margin and eyelashes, and posterior blepharitis, characterized by dysfunction of the meibomian glands. Long-term management of symptoms may include daily eyelid cleansing routines and the use of therapeutic agents that reduce infection and inflammation. A cure is not possible in most cases, and subjective symptoms may persist even when a clinical assessment of signs indicates that the condition has improved. There are no established guidelines regarding therapeutic regimens, but recent clinical trials have shown that antibiotics and topical corticosteroids can produce significant improvement in signs and symptoms of blepharitis. Fixed combinations of a topical antibiotic and a corticosteroid offer an effective and convenient treatment modality that addresses both infectious and inflammatory components of the disease. Further clinical trials are needed to determine optimal therapies for managing blepharitis.

  13. Blepharitis

    MedlinePlus

    ... Courier services use: Rockville, MD 20852) 301-451-2020 Research at NEI Office of the Scientific Director ... Eye Disease Education Program Glaucoma Education Program Low Vision Education Program Hispanic/Latino Program Vision and Aging ...

  14. [Chronic blepharitis. Pathogenesis, clinical features, and therapy].

    PubMed

    Auw-Haedrich, C; Reinhard, T

    2007-09-01

    Chronic blepharitis is one of the most common diseases of the eyelids, but surprisingly, it is not often recognized. Frequently, a skin disease such as seborrheic dermatitis, atopic dermatitis, or acne rosacea is the underlying cause of chronic blepharitis. Bacterial pathological lipase, cholesterylesterase production, and bacterial lipopolysaccharides are pathogenetically relevant. Only rarely do genuine bacterial infections play a role. Collarettes occur at the base of the eye lashes, and the Meibomian glands show either abundant fluid secretion or inspissated secretion with obstruction of the orifices. Chronic blepharitis can include sequelae including dry eye and corneal and lid contour changes. The basic treatment comprises attendance of the underlying dermatological disease and lid hygiene. In addition, preservative-free tear film substitutes, antibiotics, immunomodulatory agents, or even surgical intervention may become necessary.

  15. Blepharitis and trichomegaly induced by cetuximab.

    PubMed

    Criado, Paulo Ricardo; Lima, Aline Angélica Porto Rocha

    2010-01-01

    This report describes the case of a 41-year old woman in treatment for advanced colorectal cancer who developed severe bilateral blepharitis and trichomegaly after the second dose of cetuximab. Cetuximab-related eyelid toxicity has been described previously; however, its pathogenesis has not yet been clearly established.

  16. Acanthamoeba meningoencephalitis

    PubMed Central

    Chandra, S. R.; Adwani, Sikandar; Mahadevan, Anitha

    2014-01-01

    Report of a case of young immunocompetent male adult with autopsy proven acanthamoeba meningoencephalitis. The patient presented with a protracted febrile illness of 3 months duration with features of meningoencephalitis, this was followed by rapid deterioration while on anti tuberculous therapy and steroids and ended fatally. His magnetic resonance imaging showed features of hemorrhagic meningoencephalitis and magnetic resonance spectroscopy showed choline peak. Autopsy revealed necrotizing meningoencephalitis and intraocular colonization due to acanthamoeba. PMID:24753675

  17. [Rosacea-like demodicidosis and chronic blepharitis].

    PubMed

    Anane, S; Mokni, M; Beltaief, O

    2011-01-01

    Demodicidosis is a parasitic skin disease caused by the follicle mites Demodex sp. In this article, we present a case of rosacea-like demodicidosis, discuss the clinical features of Demodex infestation in man and review their diagnosis and therapeutic modalities. A 37-year-old woman presented in our department with chronic blepharitis present for one year. On physical examination, the patient presented blepharitis and papulovesicles with fine scaling limited to the face. There was no telangectasia. The patient did not report flushing episodes or any kind of photosensitivity. A diagnosis of rosacea-like demodicidosis and Demodex blepharitis was based on the presence of numerous Demodex folliculorum in the eyelashes and scrapings of skin lesions. The patient was put on topic and oral metronidazole for 2 months and on yellow mercury ointment for 15 days. The course involved disappearance of the facial mites and complete remission without recurrence. Screening for Demodex sp is essential to establish the correct diagnosis and ensure suitable treatment.

  18. [Acanthamoeba keratitis].

    PubMed

    Bouheraoua, N; Labbé, A; Chaumeil, C; Liang, Q; Laroche, L; Borderie, V

    2014-10-01

    Early diagnosis and appropriate therapy are key elements for a good prognosis in Acanthamoeba keratitis (AK). AK should be considered in any case of corneal trauma complicated by exposure to soil or contaminated water, and in all contact lens (CL) wearers. A presumptive diagnosis of AK can be made clinically and with in vivo confocal microscopy, although a definitive diagnosis requires identification of Acanthamoeba on direct scraping, histology, or identification of Acanthamoeba DNA by polymerase chain reaction (PCR). We use cysticidal drugs for treating AK because encysted forms are more resistant than trophozoites to treatment. The treatment protocol used a biguanide (PHMB 0.02% or chlorhexidine 0.02%) and a diamidine (propamidine 0.1% or hexamidine 0.1%). New diagnostic modalities and more specific topical anti-amoebic treatments would substantially benefit patients with AK.

  19. [Chronic blepharitis: which role for Demodex folliculorum? A case report].

    PubMed

    Martinaud, C; Gaillard, T; Pons, S; Fournier, B; Brisou, P

    2009-01-01

    We present a 73-year-old woman presented to our hospital with a 2 years history of eyes itching. The ophthalmological testing was normal. Physical examination revealed blepharitis and lesions acnea-like on mouth, nose and chest. Biological testing revealed no abnormalities. Histologic study and direct immunofluorescence on a cutaneous biopsy were no contributive. The research of an allergic origine was practised by cutaneous and serological tests and negative. An examination of eyelashes was performed and yielded Demodex. Demodex folliculorum is a mite that is the most common permanent ectoparasite of humans, which is thought to be linked to blepharitis and allergic blepharoconjunctivis with rosacea, although much controversy persists. Recent studies demonstrate a high frequence of chronic blepharitis when Demodex are abundant. Several molecules can be used to treat this infestation. Parasiticide as oral ivermectine may be useful when the infestation is important.

  20. Corneal astigmatism after cataract surgery.

    PubMed

    Dekkers, N W; Buijs, J

    1989-08-01

    206 Consecutive cataract patients were at random divided into three groups according to the way the cataract incision was closed: virgin silk 8-0, interrupted nylon 9-0, and double running nylon 9-0. The nylon, whether interrupted or continuous, yielded in the majority of cases a postoperative astigmatism with the rule, whereas virgin silk caused in nearly all patients a postoperative astigmatism against the rule and therefore behaved like an absorbable suture. Silk is chemically non-absorbable, but in virgin silk a natural worm-produced polymer is still present, which provokes a tissue reaction. Softening of tissue diminishes the tensile strength of the suture. With respect to the postoperative astigmatism, the suture material (nylon or virgin silk) seems a more important factor than the way in which it is used (interrupted or continuous).

  1. Pathogenesis of Acanthamoeba infections.

    PubMed

    Khan, Naveed Ahmed

    2003-06-01

    Acanthamoeba are free-living, harmless organisms, however, given the opportunity and the appropriate conditions, they can cause painful, sight-threatening as well as fatal infections and, thus, are considered opportunistic pathogens. Acanthamoeba infections have become increasingly important in the past few years due to increasing populations of contact lens users and AIDS patients. The mechanisms associated with the pathogenesis of Acanthamoeba tend to be highly complex, depending on parasite, host and the environmental factors. Elucidation of the biochemical, cellular and molecular basis of the pathogenesis of diseases caused by Acanthamoeba may lead to the development of therapeutic interventions.

  2. Persistence of acanthamoeba antigen following acanthamoeba keratitis

    PubMed Central

    Yang, Y; Matheson, M; Dart, J; Cree, I

    2001-01-01

    AIM—To investigate the hypothesis that persistent corneal and scleral inflammation following acanthamoeba keratitis is not always caused by active amoebic infection but can be due to persisting acanthamoebic antigens
METHODS—24 lamellar corneal biopsy and penetrating keratoplasty specimens were obtained from 14 consecutive patients at various stages of their disease and divided for microscopy and culture. Histological sections were immunostained and screened for the presence of Acanthamoeba cysts by light microscopy. Cultures were carried out using partly homogenised tissues on non-nutrient agar seeded with E coli. Clinical data were obtained retrospectively from the case notes of these patients.
RESULTS—Of the 24 specimens, 20 were obtained from eyes that were clinically inflamed at the time of surgery. Acanthamoeba cysts were present in 16 (80%) of these 20 specimens, while only five (25%) were culture positive. Acanthamoeba cysts were found to persist for up to 31 months after antiamoebic treatment.
CONCLUSION—These findings support the hypothesis that Acanthamoeba cysts can remain in corneal tissue for an extended period of time following acanthamoeba keratitis and may cause persistent corneal and scleral inflammation in the absence of active amoebic infection. In view of these findings, prolonged intensive antiamoebic therapy may be inappropriate when the inflammation is due to retained antigen rather than to viable organisms

 PMID:11222330

  3. DEBS – a unification theory for dry eye and blepharitis

    PubMed Central

    Rynerson, James M; Perry, Henry D

    2016-01-01

    For many years, blepharitis and dry eye disease have been thought to be two distinct diseases, and evaporative dry eye distinct from aqueous insufficiency. In this treatise, we propose a new way of looking at dry eye, both evaporative and insufficiency, as the natural sequelae of decades of chronic blepharitis. Dry eye is simply the late form and late manifestation of one disease, blepharitis. We suggest the use of a new term in describing this one chronic disease, namely dry eye blepharitis syndrome (DEBS). Bacteria colonize the lid margin within a structure known as a biofilm. The biofilm allows for population densities that initiate quorum-sensing gene activation. These newly activated gene products consist of inflammatory virulence factors, such as exotoxins, cytolytic toxins, and super-antigens, which are then present for the rest of the patient’s life. The biofilm never goes away; it only thickens with age, producing increasing quantities of bacterial virulence factors, and thus, increasing inflammation. These virulence factors are likely the culprits that first cause follicular inflammation, then meibomian gland dysfunction, aqueous insufficiency, and finally, after many decades, lid destruction. We suggest that there are four stages of DEBS which correlate with the clinical manifestations of folliculitis, meibomitis, lacrimalitis, and finally lid structure damage evidenced by entropion, ectropion, and floppy eyelid syndrome. When one fully understands the structure and location of the glands within the lid, it becomes easy to understand this staged disease process. The longer a gland can resist the relentless encroachment of the invading biofilm, the longer it can maintain normal function. The stages depend purely on anatomy and years of biofilm presence. Dry eye now becomes a very easy disease to understand. We feel that dry eye should be treated and prevented by early and routine biofilm removal through electromechanical lid margin debridement. PMID

  4. Use of azithromycin ophthalmic solution in the treatment of chronic mixed anterior blepharitis.

    PubMed

    John, Thomas; Shah, Ami A

    2008-01-01

    We tested the efficacy of azithromycin ophthalmic solution for the treatment of chronic mixed anterior blepharitis. The findings suggest that patients with chronic mixed anterior blepharitis can be more effectively treated with azithromycin ophthalmic solution than erythromycin ophthalmic ointment. Patients treated with azithromycin ophthalmic solution show an extraordinary clinical response with shorter treatment duration.

  5. Periodic acid–Schiff staining demonstrates fungi in chronic anterior blepharitis

    PubMed Central

    Dadaci, Z; Kılınç, F; Ozer, T T; Sahin, G O; Acir, N O; Borazan, M

    2015-01-01

    Purpose To evaluate the presence of fungi in patients with chronic anterior blepharitis with periodic acid–Schiff (PAS) staining of the eyelashes in addition to the conventional methods of fungal cultures and direct microscopy. Methods Nineteen patients with chronic anterior blepharitis of seborrheic or mixed seborrheic/staphylococcal type and 11 healthy age- and sex-matched controls were included in this prospective, nonrandomized, cross-sectional study. Blepharitis was diagnosed based on clinical evidence of greasy scales between the cilia, lid margin erythema, conjunctival hyperemia, telangiectasia, thickening, or irregularity of the eyelid margins by slit-lamp biomicroscopy. Eyelash samples were obtained by epilation with a sterile forceps and evaluated with PAS staining, fungal cultures, and direct microscopy. Results We demonstrated fungal elements with PAS staining in 79% of the blepharitis group (hyphae and/or spores) and 18% of the control group. The difference was statistically significant (P=0.002). Four patients in the blepharitis group (21%) had positive cultures for fungi. The isolated fungi were Penicillium species (2 cases), Candida species (1 case), and Trichophyton verrucosum (1 case). Direct microscopic examination revealed Demodex mites in 42.1% of the blepharitis group. No culture growth or Demodex mites were observed in the control group. Conclusions We have shown fungi with PAS staining in the majority of patients with chronic anterior blepharitis. Further controlled studies are necessary to clarify the role of fungi in the etiopathogenesis of blepharitis. PMID:26293142

  6. Detection of Acanthamoeba on the ocular surface in a Spanish population using the Schirmer strip test: pathogenic potential, molecular classification and evaluation of the sensitivity to chlorhexidine and voriconazole of the isolated Acanthamoeba strains.

    PubMed

    Rocha-Cabrera, Pedro; Reyes-Batlle, María; Martín-Navarro, Carmen María; Dorta-Gorrín, Alexis; López-Arencibia, Atteneri; Sifaoui, Ines; Martínez-Carretero, Enrique; Piñero, José E; Martín-Barrera, Fernando; Valladares, Basilio; Lorenzo-Morales, Jacob

    2015-08-01

    Pathogenic strains of Acanthamoeba are causative agents of a sight-threatening infection of the cornea known as Acanthamoeba keratitis, which is often associated with the misuse of contact lenses. However, there is still a question remaining to be answered, which is whether these micro-organisms are present on the ocular surface of healthy individuals. Therefore, the aim of this study was to determine the presence of Acanthamoeba on the ocular surface in healthy patients and also in those with other ocular surface infections. Sterile Schirmer test strips were used to collect samples from a group of patients who attended an ophthalmology consultation at the Hospital del Norte, Icod de los Vinos, Tenerife, Canary Islands. Most of the patients (46 individuals, 79.31  %) presented ocular surface pathologies such as blepharitis or conjunctivitis; the rest did not present any pathology. None of the patients included in the study wore contact lenses. The collected samples were cultured in 2  % non-nutrient agar plates and positive plates were then cultured in axenic conditions for further analyses. Molecular analysis classified all isolated strains as belonging to Acanthamoeba genotype tbl4, and osmotolerance and thermotolerance assays revealed that all strains were potentially pathogenic. Furthermore, all strains were assayed for sensitivity against voriconazole and chlorhexidine. Assays showed that both drugs were active against the tested strains. In conclusion, the Schirmer strip test is proposed as an effective tool for the detection of Acanthamoeba on the ocular surface.

  7. Comparison of incidence of Demodex folliculorum on the eyelash follicule in normal people and blepharitis patients.

    PubMed

    Türk, Meral; Oztürk, Ilgin; Sener, Asli Gamze; Küçükbay, Safiye; Afşar, Ilhan; Maden, Ahmet

    2007-01-01

    Demodex folliculorum has been incriminated in the development of blepharitis although much controversy persist. Some authors suggest that Demodex is a direct pathogen in chronic palpebral conditions while others consider the saprophyte to be innocuous to skin. We conducted a prospective study of eyelashes in 48 patients with blepharitis (totally 96 eyes) and 48 healthy persons (totally 96 eyes), searching for Demodex folliculorum and chronic blepharitis. Microscopy in immersion oil after storage in a moist chamber was performed. It is determined Demodex follicullorum in 11 of 37 (29.72%) patients with blepharitis, in one of 11 (9.09%) patient with blepharoconjunctivitis and in 2 of 48 (4.16%) persons in healthy control group. Staphylococcus aureus was cultivated in five patients and D. folliculorum was found two of them. The incidence of Demodex folliculorum was high in patients with blepharitis compared with normal controls. Incidence was high in males. Anti-Demodex treatment is indicated when the parasite is found.

  8. Astigmatism

    MedlinePlus

    ... easily diagnosed by a standard eye exam with refraction test . Special tests are not required in most ... or adults who cannot respond to a normal refraction test can have their refraction measured by a ...

  9. Azithromycin in DuraSite® for the treatment of blepharitis

    PubMed Central

    Luchs, Jodi

    2010-01-01

    Blepharitis is a common inflammatory disease of the eyelid. Posterior blepharitis affects the posterior lamella of the eyelid and involves inflammation of the meibomian glands, whereas anterior blepharitis affects the anterior lamella of the eyelid and the eyelashes; either version can be inflammatory or infectious in nature. Each of these conditions can incite or propagate the other; anterior blepharitis, if not treated, can lead to meibomian gland disease, and vice versa. Blepharitis is typically chronic, and can be associated with a variety of systemic diseases such as dermatitis, as well as ocular diseases such as dry eye, conjunctivitis, or keratitis. The standard treatment regimen historically consists of lid hygiene with warm compresses and eyelid scrubs, although these treatment modalities may have limited efficacy for many patients, especially those with more severe disease. Adjunctive treatment includes systemic and topical antibiotics, topical corticosteroids, and tear replacement therapy. Topical antibiotics are recommended to decrease the bacterial load, and topical corticosteroids may help in cases of severe inflammation. Azithromycin ophthalmic solution 1% in DuraSite® (AzaSite®; Inspire Pharmaceuticals, Durham, North Carolina, USA) has been proposed as a novel treatment for posterior blepharitis, based on its well-known anti-infective profile, its anti-inflammatory properties, its excellent tissue penetration, and its regulatory approval for the treatment of bacterial conjunctivitis. This review focuses on an off-label indication for topical azithromycin 1% in DuraSite for the treatment of blepharitis. PMID:20689782

  10. Blepharitis in the United States 2009: a survey-based perspective on prevalence and treatment.

    PubMed

    Lemp, Michael A; Nichols, Kelly K

    2009-04-01

    ABSTRACT Like dry eye disease 15 years ago, blepharitis today is a poorly defined condition about which there is considerable misunderstanding. For a variety of reasons, there is little good data on either the prevalence of blepharitis or how eyecare practitioners currently treat it. The work reported herein consists of two recent studies: a telephone survey of a representative sample of the adult US population (n = 5,000) whose purpose was to discover the frequency of common ocular surface symptoms associated with blepharitis; and a study that queried a selected group of ophthalmologists (n = 120) and a similarly selected group of optometrists (n = 84) about the frequency of blepharitis in their practices, the existence comorbid conditions, and their management strategies. This data suggests that blepharitis symptoms are very common in the US population, with younger individuals reporting more, and more frequent, symptoms than older people, contrary to clinical dogma. Ophthalmologists and optometrists report that blepharitis is commonly seen in clinical practice in 37% and 47% of their patients, respectively, and it is widely agreed that meibomian gland dysfunction (MGD) is the most common cause of evaporative dry eye disease. In addition, management paradigms are shifting away from more traditional management with antibiotic ointment and warm compress therapy to prescription therapy for anterior and posterior blepharitis.

  11. Acanthamoeba keratitis in Pondicherry.

    PubMed

    Parija, S C; Prakash, M R; Rao, V A; Vellaniparambil, R J

    2001-06-01

    Acanthamoeba keratitis is a potentially devastating infection of the cornea caused by the free-living amoebae, Acanthamoeba species. During the period from 1997 to 2000, a total of 136 corneal scrapings from clinically suspected cases were screened and examined for the presence of the Acanthamoeba. On examination of the direct smear by microscopy, 11 out of 136 cases were positive for Acanthamoeba. Eight patients were males and 3 were females. The age of these patients ranged from 15 to 57 years. All of these cases were agricultural workers who did not use contact lens. Four cases gave a history of injury to the eye and 1 patient gave a history of applying cow dung on the eye after the injury. Rest of the patients did not give any history of trauma or wearing contact lenses. The patients were treated with topical application of neosporin ointment. Many of our cases had complications such as poor vision (all 11 cases had 6/60 or less), scar formation (3 cases), opacity (5 cases) and corneal perforation (2 cases). This report documents for the first time the cases of Acanthamoeba keratitis in Pondicherry.

  12. Radial averages of astigmatic TEM images.

    PubMed

    Fernando, K Vince

    2008-10-01

    The Contrast Transfer Function (CTF) of an image, which modulates images taken from a Transmission Electron Microscope (TEM), is usually determined from the radial average of the power spectrum of the image (Frank, J., Three-dimensional Electron Microscopy of Macromolecular Assemblies, Oxford University Press, Oxford, 2006). The CTF is primarily defined by the defocus. If the defocus estimate is accurate enough then it is possible to demodulate the image, which is popularly known as the CTF correction. However, it is known that the radial average is somewhat attenuated if the image is astigmatic (see Fernando, K.V., Fuller, S.D., 2007. Determination of astigmatism in TEM images. Journal of Structural Biology 157, 189-200) but this distortion due to astigmatism has not been fully studied or understood up to now. We have discovered the exact mathematical relationship between the radial averages of TEM images with and without astigmatism. This relationship is determined by a zeroth order Bessel function of the first kind and hence we can exactly quantify this distortion in the radial averages of signal and power spectra of astigmatic images. The argument to this Bessel function is similar to an aberration function (without the spherical aberration term) except that the defocus parameter is replaced by the differences of the defoci in the major and minor axes of astigmatism. The ill effects due this Bessel function are twofold. Since the zeroth order Bessel function is a decaying oscillatory function, it introduces additional zeros to the radial average and it also attenuates the CTF signal in the radial averages. Using our analysis, it is possible to simulate the effects of astigmatism in radial averages by imposing Bessel functions on idealized radial averages of images which are not astigmatic. We validate our theory using astigmatic TEM images.

  13. Visual Motor and Perceptual Task Performance in Astigmatic Students.

    PubMed

    Harvey, Erin M; Twelker, J Daniel; Miller, Joseph M; Leonard-Green, Tina K; Mohan, Kathleen M; Davis, Amy L; Campus, Irene

    2017-01-01

    Purpose. To determine if spectacle corrected and uncorrected astigmats show reduced performance on visual motor and perceptual tasks. Methods. Third through 8th grade students were assigned to the low refractive error control group (astigmatism < 1.00 D, myopia < 0.75 D, hyperopia < 2.50 D, and anisometropia < 1.50 D) or bilateral astigmatism group (right and left eye ≥ 1.00 D) based on cycloplegic refraction. Students completed the Beery-Buktenica Developmental Test of Visual Motor Integration (VMI) and Visual Perception (VMIp). Astigmats were randomly assigned to testing with/without correction and control group was tested uncorrected. Analyses compared VMI and VMIp scores for corrected and uncorrected astigmats to the control group. Results. The sample included 333 students (control group 170, astigmats tested with correction 75, and astigmats tested uncorrected 88). Mean VMI score in corrected astigmats did not differ from the control group (p = 0.829). Uncorrected astigmats had lower VMI scores than the control group (p = 0.038) and corrected astigmats (p = 0.007). Mean VMIp scores for uncorrected (p = 0.209) and corrected astigmats (p = 0.124) did not differ from the control group. Uncorrected astigmats had lower mean scores than the corrected astigmats (p = 0.003). Conclusions. Uncorrected astigmatism influences visual motor and perceptual task performance. Previously spectacle treated astigmats do not show developmental deficits on visual motor or perceptual tasks when tested with correction.

  14. Visual Motor and Perceptual Task Performance in Astigmatic Students

    PubMed Central

    Twelker, J. Daniel; Miller, Joseph M.; Mohan, Kathleen M.; Campus, Irene

    2017-01-01

    Purpose. To determine if spectacle corrected and uncorrected astigmats show reduced performance on visual motor and perceptual tasks. Methods. Third through 8th grade students were assigned to the low refractive error control group (astigmatism < 1.00 D, myopia < 0.75 D, hyperopia < 2.50 D, and anisometropia < 1.50 D) or bilateral astigmatism group (right and left eye ≥ 1.00 D) based on cycloplegic refraction. Students completed the Beery-Buktenica Developmental Test of Visual Motor Integration (VMI) and Visual Perception (VMIp). Astigmats were randomly assigned to testing with/without correction and control group was tested uncorrected. Analyses compared VMI and VMIp scores for corrected and uncorrected astigmats to the control group. Results. The sample included 333 students (control group 170, astigmats tested with correction 75, and astigmats tested uncorrected 88). Mean VMI score in corrected astigmats did not differ from the control group (p = 0.829). Uncorrected astigmats had lower VMI scores than the control group (p = 0.038) and corrected astigmats (p = 0.007). Mean VMIp scores for uncorrected (p = 0.209) and corrected astigmats (p = 0.124) did not differ from the control group. Uncorrected astigmats had lower mean scores than the corrected astigmats (p = 0.003). Conclusions. Uncorrected astigmatism influences visual motor and perceptual task performance. Previously spectacle treated astigmats do not show developmental deficits on visual motor or perceptual tasks when tested with correction. PMID:28293434

  15. Bacillus oleronius and Demodex mite infestation in patients with chronic blepharitis.

    PubMed

    Szkaradkiewicz, A; Chudzicka-Strugała, I; Karpiński, T M; Goślińska-Pawłowska, O; Tułecka, T; Chudzicki, W; Szkaradkiewicz, A K; Zaba, R

    2012-10-01

    To better recognize the pathogenicity of ocular Demodex mites, we analysed Bacillus oleronius infection in patients with Demodex-related chronic blepharitis. The studies were conducted on 68 adult patients, in whom ophthalmological and parasitological tests permitted the distinction of a group of 38 patients with a diagnosis of Demodex-related chronic blepharitis (group 1, including a subgroup 1a with moderate blepharitis and a subgroup 1b with severe blepharitis) and a group of 30 healthy individuals (group 2). In every person studied six eyelashes were epilated from each eye and the number of Demodex per eyelash was scored. In parallel, bacterial culture and isolation allowed their phenotypic and molecular identification. The drug sensitivity of the isolates was tested using E-tests. Intensity of Demodex infestation showed no significant differences between subgroups 1a and 1b. From the epilated eyelashes 23 bacterial isolates were obtained, identified as being B. oleronius. All the studied strains were sensitive to ciprofloxacin, doxycycline and gentamicin. The Demodex mite represents an independent aetiopathogenetic factor in blepharitis. In parallel, the parasite may act as a carrier of B. oleronius bacteria, which most probably function as a co-pathogen in the development of severe forms of blepharitis.

  16. Comparative analysis of the tear protein expression in blepharitis patients using two-dimensional electrophoresis.

    PubMed

    Koo, Bon-Suk; Lee, Do-Yeon; Ha, Hyo-Shin; Kim, Jae-Chan; Kim, Chan-Wha

    2005-01-01

    Change in the expression of body fluid proteins is caused by many diseases or environmental disturbances. The changes in tear proteins are also associated with various pathological eye conditions. Especially, chronic blepharitis is one of the most common conditions seen in the ophthalmologist's office. However, there are no specific clinical diagnostic tests for blepharitis, and it is difficult to treat effectively. Therefore, the aim of this study was to screen prognostic or diagnostic marker tear proteins for blepharitis and investigate pathogenesis of this disease using proteomics techniques. The tear proteins expressed in patients suffering from blepharitis (patient, n=19) and healthy volunteers (control, n=27) were analyzed using the two-dimensional electrophoresis (2-DE) technique. The differentially expressed proteins in patients were identified with ESI-Q-TOF (electrospray-quadrupole-time-of-flight) mass spectrometry and confirmed with western blotting. Nine proteins in patient were down regulated about 50% compared to those of the control: serum albumin precursor, alpha-1 antitrypsin, lacritin precursor, lysozyme, Ig-kappa chain VIII, prolactin inducible protein (PIP/GCDFP-15), cystatin-SA III, pyruvate kinase, and an unnamed protein. The use of the two-dimensional eletrophoretic technique could give more insight into the disease-related protein expression changes in tear fluids. Our findings reveal that the composition of tear proteins in blepharitis patients is different from that of healthy subjects and may provide further insights into the pathogenesis of blepharitis.

  17. Biology and pathogenesis of Acanthamoeba.

    PubMed

    Siddiqui, Ruqaiyyah; Khan, Naveed Ahmed

    2012-01-10

    Acanthamoeba is a free-living protist pathogen, capable of causing a blinding keratitis and fatal granulomatous encephalitis. The factors that contribute to Acanthamoeba infections include parasite biology, genetic diversity, environmental spread and host susceptibility, and are highlighted together with potential therapeutic and preventative measures. The use of Acanthamoeba in the study of cellular differentiation mechanisms, motility and phagocytosis, bacterial pathogenesis and evolutionary processes makes it an attractive model organism. There is a significant emphasis on Acanthamoeba as a Trojan horse of other microbes including viral, bacterial, protists and yeast pathogens.

  18. Two cases of tinea ciliaris with blepharitis due to Microsporum audouinii and Trichophyton verrucosum and review of the literature.

    PubMed

    Sahin, Gulay Ozel; Dadaci, Zeynep; Ozer, Turkan Toka

    2014-09-01

    Dermatophytes are rarely taken into account among the causes of blepharitis. In our report, we describe a 69-year-old man and a 40-year-old woman with chronic blepharitis for 10 years and 4 years respectively, in whom we examined the scales and pulled eyelashes on direct microscopy and isolated Microsporum audouinii and Trichophyton verrrucosum in the culture. We emphasise that dermatophytes may play a role in the etiopathogenesis of chronic blepharitis. In chronic, treatment resistance blepharitis fungal infections may be considered as possible cause.

  19. Reading Fluency in School-Age Children with Bilateral Astigmatism

    PubMed Central

    Harvey, Erin M.; Miller, Joseph M.; Twelker, J. Daniel; Davis, Amy L.

    2015-01-01

    Purpose To compare oral reading fluency (ORF) in students with no/low astigmatism and moderate/high astigmatism and to assess the impact of spectacle correction on ORF in moderate and high astigmats. Methods Subjects were 3rd–8th grade students from a highly astigmatic population. Refractive error was determined through subjectively refined cycloplegic autorefraction. Data from students with ocular abnormalities, anisometropia, symptomatic binocular vision disorders, or refractive error that did not meet study criteria (no/low (cyl < 1.00 both eyes, no significant myopia/hyperopia), moderate (cyl ≥ 1.00 D both eyes, mean ≥ 1.00 D and < 3.00 D), or high astigmatism group (cyl ≥ 1.00 D both eyes, mean ≥ 3.00 D)) were excluded. ORF was tested with a modified version of the DIBELS Next test of ORF. No/low astigmats were tested without spectacles; astigmats were tested with and without spectacles. Mean ORF was compared in no/low astigmats and astigmats (with and without correction). Improvement in ORF with spectacles was compared for moderate and high astigmats. Results The sample included 130 no/low, 67 moderate, and 76 high astigmats. ORF was lower in uncorrected astigmats than in no/low astigmats (p=0.011). ORF did not significantly differ in no/low astigmats and corrected astigmats (p=0.10). ORF significantly improved with spectacle correction in high astigmats (p=0.001, mean improvement 6.55 words per minute (WPM)), but not in moderate astigmats (p=0.193, mean improvement 1.87 WPM). Effects of spectacle wear were observed in students who read smaller text stimuli (older grades). Conclusions ORF is significantly reduced in students with bilateral astigmatism (≥ 1.00D) when uncorrected, but not when best-corrected, compared to their non-astigmatic peers. Improvement in ORF with spectacle correction is seen in high astigmats, but not in moderate astigmats. These data support the recommendation for full-time spectacle wear in astigmatic students

  20. [Essential features of astigmatism and its correction with excimer laser].

    PubMed

    Vlaicu, Valeria

    2012-01-01

    The correction of astigmatism is an essential element for the refractive surgery because the majority of patients have important preoperative cylinder An uncorrected astigmatism decreases visual acuity and can also cause glare, asthenopia, headaches, monocular diplopia. It is important to remark that a complete elimination of astigmatism for the eye is very rarely achieved.

  1. Optical advantages of astigmatic aberration corrected heliostats

    NASA Astrophysics Data System (ADS)

    van Rooyen, De Wet; Schöttl, Peter; Bern, Gregor; Heimsath, Anna; Nitz, Peter

    2016-05-01

    Astigmatic aberration corrected heliostats adapt their shape in dependence of the incidence angle of the sun on the heliostat. Simulations show that this optical correction leads to a higher concentration ratio at the target and thus in a decrease in required receiver aperture in particular for smaller heliostat fields.

  2. Ocular rosacea: an underdiagnosed cause of relapsing conjunctivitis-blepharitis in the elderly.

    PubMed

    De Marchi, Sergio Umberto; Cecchin, Emanuela; De Marchi, Sergio

    2014-09-19

    Red eye and relapsing conjunctivitis-blepharitis are among the most common ocular disease in elderly patients. In these cases the search for causes is difficult and frustrating. We report the case of a 79-year-old woman with a long history of red eye and relapsing conjunctivitis-blepharitis caused by ocular rosacea. In this patient the proper diagnosis was performed after 10 years of ocular disease, and repeated evaluations by general practitioners and clinical specialists, only after the appearance of facial signs of erythematotelangiectatic rosacea. Adequate therapy with oral doxycycline led to the improvement of the clinical picture that previously had shown a poor response to several topical treatments. The possibility of ocular rosacea should be considered in evaluating an elderly patient with persistent red eye and relapsing conjunctivitis-blepharitis. Making the proper diagnosis is crucial because ocular rosacea does not respond as expected to topical therapy and may lead to severe corneal involvement.

  3. Adaptive immune responses to Acanthamoeba cysts.

    PubMed

    McClellan, Kathy; Howard, Kevin; Mayhew, Elizabeth; Niederkorn, Jerry; Alizadeh, Hassan

    2002-09-01

    Acanthamoeba cysts are not eliminated from the corneas of human subjects or experimentally infected animals. The persistence of Acanthamoeba cysts in the cornea indicates that either the cysts escape immunological elimination or are not recognized by the host's immunological elements. The aim of this study was to determine the immunogenicity and antigenicity of the Acanthamoeba cyst. Mice were immunized intraperitoneally and serum anti-Acanthamoeba IgG was measured by ELISA. Lymphoproliferative assay and delayed type hypersensitivity (DTH) responses to Acanthamoeba castellanii cyst and trophozoite antigens were used to determine the cell mediated immune responses against Acanthamoeba cysts. A. castellanii cysts were both immunogenic and antigenic, producing anti-Acanthamoeba serum IgG, T lymphocyte proliferation, and delayed type hypersensitivity responses. These results indicate that Acanthamoeba cysts are recognized by the immune system. The persistence of the organism in the human cornea means that these adaptive immune responses fail to kill Acanthamoeba cysts.

  4. Prevalence of corneal astigmatism before cataract surgery.

    PubMed

    Mohammadi, Mehran; Naderan, Mohammad; Pahlevani, Rozhin; Jahanrad, Ali

    2016-12-01

    The purpose of this study was to describe and analyze the prevalence and pattern of corneal astigmatism in cataract surgery candidates. In a prospective cross-sectional study, preoperative demographics, and keratometric and refractive values of cataract surgery candidates were collected from January 2013 to December 2014. Axial length (AL) and flat and steep keratometry measurements were optically measured by a partial coherence interferometry device (IOLMaster). This study consisted of 2156 eyes of 1317 patients with a mean age of 64.92 ± 11.48 (SD) (30-88 years). The mean of AL was 23.33 ± 1.37 mm, and the mean of corneal astigmatism was 1.12 ± 1.10 diopter (D) (range 0.0-7.00), in all patients. Furthermore, the mean of flat and steep keratometry were 43.70 ± 1.70 and 44.83 ± 1.79 D, respectively. Corneal astigmatism was 1.50 D or less in 1590 eyes (73.7 %), more than 1.50 D in 566 eyes (26.2 %), 3.00 D or more in 161 eyes (7.4 %), WTR in 796 eyes (36.9 %), ATR in 1010 eyes (46.8 %), and oblique in 350 eyes (16.2 %). ATR astigmatism axis significantly increased with the increase in age. Corneal astigmatism of most cataract surgery candidates fell between 0.50 and 1.50 D. The results of our study however is confined to our demographics might provide useful data for cataract patients, surgeons, and intraocular lens manufacturers for different purposes.

  5. Treatment of Acanthamoeba keratitis.

    PubMed

    Seal, David

    2003-08-01

    The treatment of Acanthamoeba keratitis has now been possible since the first successful therapy developed in the mid 1980s with a combination of propamidine 0.1% (Brolene) and neomycin 1%. However, only half the patients responded to this regimen as the cysts were often resistant to neomycin and relatively insensitive to propamidine. This led to research for better therapy, culminating in the mid 1990s with research in Glasgow demonstrating much increased effectiveness with use of the biguanide chlorhexidine 0.02% and in London and Bristol for similar effectiveness with the polymeric polyhexamethylene biguanide (PHMB) 0.02%. Both biguanides were combined with propamidine for enhanced effectiveness but were also shown to be effective as monotherapy. While this therapy inactivates the trophozoites and cysts in Acanthamoeba keratitis in the majority of patients (approximately 90%), there have been notable failures particularly when presentation is late with deep stromal infection. Additional highly acanthamoebicidal drugs are needed that can penetrate the stroma for synergistic action. This role may be taken up by certain antineoplastic drugs, such as alkylphosphocholine-1 (Miltefosine), that also have antiprotozoal activity.

  6. Rays and fields in general astigmatic resonators

    NASA Astrophysics Data System (ADS)

    Weber, H.

    2012-05-01

    General astigmatic (GA) resonators are discussed in detail. Eigenrays, eigenmodes and eigenvalues (Gouy-factors) of this resonator are evaluated. A stability diagram for such resonators is introduced, which clearly depicts the stable and unstable regions for rays as well as for fields. Eigenrays and their stability regions are evaluated using the ABCD-law. For the beam propagation Collins' integral and the second moment method are applied. The eigenfunctions for rectangular symmetry are the generalized Hermite polynomials multiplied by the Gaussian exponential factor. It is shown that for general astigmatic resonators these polynomials are the product of normal Hermite polynomials. The generating function of the eigenfunctions depends on the special resonator. It is a useful tool for all calculations and it is determined. Furthermore it is shown that the second moment characterization of the modes is a useful and easy to handle procedure to evaluate beam width, beam divergence, radius of curvature and twist of the generalized Gauss-Hermite functions.

  7. Effects of posterior corneal astigmatism on the accuracy of AcrySof toric intraocular lens astigmatism correction

    PubMed Central

    Zhang, Bin; Ma, Jing-Xue; Liu, Dan-Yan; Guo, Cong-Rong; Du, Ying-Hua; Guo, Xiu-Jin; Cui, Yue-Xian

    2016-01-01

    AIM To evaluate the effects of posterior corneal surface measurements on the accuracy of total estimated corneal astigmatism. METHODS Fifty-seven patients with toric intraocular lens (IOL) implantation and posterior corneal astigmatism exceeding 0.5 diopter were enrolled in this retrospective study. The keratometric astigmatism (KA) and total corneal astigmatism (TA) were measured using a Pentacam rotating Scheimpflug camera to assess the outcomes of AcrySof IOL implantation. Toric IOLs were evaluated in 26 eyes using KA measurements and in 31 eyes using TA measurements. Preoperative corneal astigmatism and postoperative refractive astigmatism were recorded for statistical analysis. The cylindrical power of toric IOLs was estimated in all eyes. RESULTS In all cases, the difference of toric IOL astigmatism magnitude between KA and TA measurements for the estimation of preoperative corneal astigmatism was statistically significant. Of a total of 57 cases, the 50.88% decreased from Tn to Tn-1, and 10.53% decreased from Tn to Tn-2. In all cases, 5.26% increased from Tn to Tn+1. The mean postoperative astigmatism within the TA group was significantly lower than that in the KA group. CONCLUSION The accuracy of total corneal astigmatism calculations and the efficacy of toric IOL correction can be enhanced by measuring both the anterior and posterior corneal surfaces using a Pentacam rotating Scheimpflug camera. PMID:27672591

  8. Optimization of astigmatic particle tracking velocimeters

    NASA Astrophysics Data System (ADS)

    Rossi, Massimiliano; Kähler, Christian J.

    2014-09-01

    Astigmatic particle tracking velocimetry (APTV) has been developed in the last years to measure the three-dimensional displacement of tracer particles using a single-camera view. The measurement principle relies on an astigmatic optical system that provides aberrated particle images with a characteristic elliptical shape univocally related to the corresponding particle depth position. Because of the precision of this method, this concept is well established for measuring and controlling the distance between a CD/DVD and the reading head. The optical arrangement of an APTV system essentially consists of a primary stigmatic optics (e.g., a microscope, or a camera objective) and an astigmatic optics, typically a cylindrical lens placed in front of the camera sensor. This paper focuses on the uncertainty of APTV in the depth direction. First, an approximated analytical model is derived and experimentally validated. From the model, a set of three non-dimensional parameters that are the most significant in the optimization of the APTV performance are identified. Finally, the effect of different parameter settings and calibration approaches are studied systematically using numerical Monte Carlo simulations. The results allow for the derivation of general criteria to minimize the overall error in APTV measurements and provide the basis for reliable uncertainty estimation for a wide range of applications.

  9. Axially astigmatic surfaces: different types and their properties

    NASA Astrophysics Data System (ADS)

    Malacara-Doblado, Daniel; Malacara-Hernandez, Daniel; Garcia-Marquez, Jorge L.

    1996-12-01

    Axially astigmatic surfaces have different curvatures in orthogonal diameters. Toroidal and spherocylindrical optical surfaces are two mathematically different special cases of axially astigmatic surfaces as noted by Menchaca and Malacara (1986), but they are almost identical in the vicinity of the optical axis. The different between these two surfaces increases when the distance to the optical axis increases. We study the general properties of astigmatic surfaces and some special interesting cases.

  10. Photochemotherapeutic strategies against Acanthamoeba keratitis

    PubMed Central

    2012-01-01

    Here, we determined the potential of photochemotherapy, namely the application of photodynamic compounds followed by exposure to a suitable source of UV-visible radiation against corneal pathogen, Acanthamoeba. Organometallic macromolecule, tin porphyrin [Sn(IV)porphyrin] was synthesized and purity confirmed using nuclear magnetic resonance spectroscopy. The Sn(IV)porphyrin was tested against a keratitis isolate of Acanthamoeba castellanii belonging to the T4 genotype using growth and viability assays. The effects of Sn(IV)porphyrin on A. castellanii binding to and cytopathogenicity of human corneal epithelial cells in vitro were tested. The metalloporphyrin showed potent amoebistatic effects. The tin porphyrin inhibited amoebae binding to and cytopathogenicity of corneal epithelial cells. By using derivatives of photodynamic compounds [Sn(IV)porphyrin-antibody conjugates] for selective targeting of the parasite together with appropriate selection of light source will determine the potential of photochemotherapy against Acanthamoeba keratitis. PMID:22950898

  11. PINOCYTOSIS IN ACANTHAMOEBA CASTELLANII

    PubMed Central

    Bowers, Blair; Olszewski, Thomas E.

    1972-01-01

    The uptake of radioactively labeled albumin, inulin, leucine, and glucose by Acanthamoeba castellanii (Neff strain) was measured. The uptake is linear with time and appears to be continuous under the conditions of these experiments. Uptake is abolished at 0°C. No evidence for saturation of the uptake mechanism was obtained with either albumin or leucine. Each of the four tracer molecules enters the ameba at a similar rate when the uptake is calculated as volume of fluid ingested per unit time. The data suggest that each of these molecules enters the cell by pinocytosis. The highest rate of uptake was obtained with cells in their usual culture medium containing proteose peptone, glucose, and salts but pinocytosis also continued at a reduced rate in a simple salt solution. The calculated volume of fluid taken in during pinocytosis in culture medium was about 2 µl/hr per 106 cells. The route of uptake was examined in the electron microscope using horseradish peroxidase (HRP) as a tracer. HRP activity was found exclusively within membrane profiles within the cytoplasm, confirming the pinocytotic mode of uptake. An estimate of the rate of surface membrane turnover due to pinocytosis was made using the biochemical and morphological data obtained. This estimate suggests that the plasma membrane turnover of one cell is on the order of several times an hour. PMID:5028259

  12. Acanthamoeba castellanii STAT Protein

    PubMed Central

    Kicinska, Anna; Leluk, Jacek; Jarmuszkiewicz, Wieslawa

    2014-01-01

    STAT (signal transducers and activators of transcription) proteins are one of the important mediators of phosphotyrosine-regulated signaling in metazoan cells. We described the presence of STAT protein in a unicellular, free-living amoebae with a simple life cycle, Acanthamoeba castellanii. A. castellanii is the only, studied to date, Amoebozoan that does not belong to Mycetozoa but possesses STATs. A sequence of the A. castellanii STAT protein includes domains similar to those of the Dictyostelium STAT proteins: a coiled coil (characteristic for Dictyostelium STAT coiled coil), a STAT DNA-binding domain and a Src-homology domain. The search for protein sequences homologous to A. castellanii STAT revealed 17 additional sequences from lower eukaryotes. Interestingly, all of these sequences come from Amoebozoa organisms that belong to either Mycetozoa (slime molds) or Centramoebida. We showed that there are four separated clades within the slime mold STAT proteins. The A. castellanii STAT protein branches next to a group of STATc proteins from Mycetozoa. We also demonstrate that Amoebozoa form a distinct monophyletic lineage within the STAT protein world that is well separated from the other groups. PMID:25338074

  13. Acanthamoeba everywhere: high diversity of Acanthamoeba in soils.

    PubMed

    Geisen, Stefan; Fiore-Donno, Anna Maria; Walochnik, Julia; Bonkowski, Michael

    2014-09-01

    Acanthamoeba is a very abundant genus of soil protists with fundamental importance in nutrient cycling, but several strains can also act as human pathogens. The systematics of the genus is still unclear: currently 18 small-subunit (SSU or 18S) ribosomal RNA sequence types (T1-T18) are recognized, which sometimes contain several different morphotypes; on the other hand, some morphological identical strains belong to different sequence types, sometimes appearing in paraphyletic positions. In this study, we cultivated 65 Acanthamoeba clones from soil samples collected under grassland at three separate locations in the Netherlands, in Sardinia and at high altitude mountains in Tibet. We obtained 24 distinct partial sequences, which predominantly grouped within sequence type T4 followed by T2, T13, T16 and "OX-1" (in the T2/T6 clade). Our sequences were 98-99% similar, but none was identical to already known Acanthamoeba sequences. The community composition of Acanthamoeba strains differed between locations, T4 being the dominant sequence type in Sardinia and Tibet, but represented only half of the clones from soils in the Netherlands. The other half of clones from the Dutch soils was made up by T2, T16 and "OX-1", while T13 was only found in Sardinia and Tibet. None of the sequences was identical between localities. Several T4 clones from all three localities and all T13 clones grew at 37 °C while one T4 clone was highly cytopathogenic.

  14. Lack of oblique astigmatism in the chicken eye.

    PubMed

    Maier, Felix M; Howland, Howard C; Ohlendorf, Arne; Wahl, Siegfried; Schaeffel, Frank

    2015-04-01

    Primate eyes display considerable oblique off-axis astigmatism which could provide information on the sign of defocus that is needed for emmetropization. The pattern of peripheral astigmatism is not known in the chicken eye, a common model of myopia. Peripheral astigmatism was mapped out over the horizontal visual field in three chickens, 43 days old, and in three near emmetropic human subjects, average age 34.7years, using infrared photoretinoscopy. There were no differences in astigmatism between humans and chickens in the central visual field (chicks -0.35D, humans -0.65D, n.s.) but large differences in the periphery (i.e. astigmatism at 40° in the temporal visual field: humans -4.21D, chicks -0.63D, p<0.001, unpaired t-test). The lack of peripheral astigmatism in chicks was not due to differences in corneal shape. Perhaps related to their superior peripheral optics, we found that chickens had excellent visual performance also in the far periphery. Using an automated optokinetic nystagmus paradigm, no difference was observed in spatial visual performance with vision restricted to either the central 67° of the visual field or to the periphery beyond 67°. Accommodation was elicited by stimuli presented far out in the visual field. Transscleral images of single infrared LEDs showed no sign of peripheral astigmatism. The chick may be the first terrestrial vertebrate described to lack oblique astigmatism. Since corneal shape cannot account for the difference in astigmatism in humans and chicks, it must trace back to the design of the crystalline lens. The lack of peripheral astigmatism in chicks also excludes a role in emmetropization.

  15. Accommodation in Astigmatic Children During Visual Task Performance

    PubMed Central

    Harvey, Erin M.; Miller, Joseph M.; Apple, Howard P.; Parashar, Pavan; Twelker, J. Daniel; Crescioni, Mabel; Davis, Amy L.; Leonard-Green, Tina K.; Campus, Irene; Sherrill, Duane L.

    2014-01-01

    Purpose. To determine the accuracy and stability of accommodation in uncorrected children during visual task performance. Methods. Subjects were second- to seventh-grade children from a highly astigmatic population. Measurements of noncycloplegic right eye spherical equivalent (Mnc) were obtained while uncorrected subjects performed three visual tasks at near (40 cm) and distance (2 m). Tasks included reading sentences with stimulus letter size near acuity threshold and an age-appropriate letter size (high task demands) and viewing a video (low task demand). Repeated measures ANOVA assessed the influence of astigmatism, task demand, and accommodative demand on accuracy (mean Mnc) and variability (mean SD of Mnc) of accommodation. Results. For near and distance analyses, respectively, sample size was 321 and 247, mean age was 10.37 (SD 1.77) and 10.30 (SD 1.74) years, mean cycloplegic M was 0.48 (SD 1.10) and 0.79 diopters (D) (SD 1.00), and mean astigmatism was 0.99 (SD 1.15) and 0.75 D (SD 0.96). Poor accommodative accuracy was associated with high astigmatism, low task demand (video viewing), and high accommodative demand. The negative effect of accommodative demand on accuracy increased with increasing astigmatism, with the poorest accommodative accuracy observed in high astigmats (≥3.00 D) with high accommodative demand/high hyperopia (1.53 D and 2.05 D of underaccommodation for near and distant stimuli, respectively). Accommodative variability was greatest in high astigmats and was uniformly high across task condition. No/low and moderate astigmats showed higher variability for the video task than the reading tasks. Conclusions. Accuracy of accommodation is reduced in uncorrected children with high astigmatism and high accommodative demand/high hyperopia, but improves with increased visual task demand (reading). High astigmats showed the greatest variability in accommodation. PMID:25103265

  16. Bilateral upper and lower eyelid severe psoriasiform blepharitis: case report and review of literature.

    PubMed

    Zhu, Feilin; Tao, Jeremiah P

    2011-01-01

    The purpose of the present study is to describe a case of severe, psoriasiform blepharitis by means of a case report and literature review. A 44-year-old man developed chronic blepharitis and tearing months after bilateral cataract surgery. Exam showed diffuse quad-eyelid erythema, discharge, edema, madarosis, and scale. He also had insufficient tear drainage due to bilateral upper eyelid cicatricial punctal atresia with bilateral lower eyelid punctal stenosis. Biopsy of the lower eyelids exhibited psoriasiform hyperplasia. Topical 0.1% tacrolimus achieved improvement but caused some subjective eye irritation. Psoriasiform dermatitis manifesting on the eyelids is rare, may be associated with insufficient tear drainage, and may respond favorably to 0.1% tacrolimus.

  17. A unified paraxial approach to astigmatic optics.

    PubMed

    Harris, W F

    1999-07-01

    In Gaussian optics properties such as dioptric power, lateral and angular magnification and thickness are simple scalar concepts. In linear optics, the optics of thick astigmatic systems, however, these concepts generalize to three-dimensional concepts in some cases (the dioptric power of thin systems, for example) and to four-dimensional concepts in general. As a result, the quantitative treatment of these properties in astigmatic systems presents challenges to the researcher in optometry, ophthalmology, and vision science. Considerable progress has been made only in the case of dioptric power. This paper presents a generalized approach to astigmatic optics which allows different physical properties to be treated in the same way: the theory is unified and, in a sense, complete. Mathematical and statistical methods developed for treating one concept become directly applicable to others. The paraxial optical properties of any optical system are completely defined by the 4 x 4 ray transfer matrix, called here the (ray) transference. The transference defines four fundamental properties of an optical system, tentatively called here positional magnification, optical thickness, divergence, and directional magnification. They are the four 2 x 2 submatrices A, B, C, and D of the transference. Each fundamental property is a modification of a familiar concept. Divergence is the negative of dioptric power expressed as the dioptric power matrix F. The four fundamental optical properties A, B, C, and D, and the derived property F, despite being different physically, all have the same underlying mathematical structure. This fact is exploited in developing a unified theory. The theory is complete in the sense that the fundamental properties fully characterize the paraxial optics of any system. The paper presents a general treatment that applies to any of the five properties. The implications are far reaching and extend beyond what can be described in the paper. Dioptric power of

  18. Staphylococcus aureus Blepharitis Associated with Multiple Corneal Stromal Microabscess, Stromal Edema, and Uveitis.

    PubMed

    Boto-de-los-Bueis, Ana; del Hierro Zarzuelo, Almudena; García Perea, Adela; de Pablos, Manuela; Pastora, Natalia; Noval, Susana

    2015-04-01

    We report a case of an immunocompetent woman with atypical marginal keratitis. She presented with recurrent episodes of multiples microabscess distributed in a triangular pattern associated with stromal oedema and anterior chamber uveitis, affecting both eyes, but not simultaneously. The episodes responded to steroid drops, corneal inflammation was coincidental with a worsening of her blepharitis in the affected eye and S. aureus was isolated from the lids.

  19. Comparison of therapeutic effects of topical azithromycin solution and systemic doxycycline on posterior blepharitis

    PubMed Central

    Zandian, Mehdi; Rahimian, Neda; Soheilifar, Sanaz

    2016-01-01

    AIM To compare the effect of azithromycin drop and doxycycline capsule on treatment of posterior blepharitis. METHODS Fifty patients (100 eyes) with moderate posterior blepharitis, randomly divided into two therapeutic groups; all the patients got warm eyelid compress and massage three times a day for 3wk. In addition the first group got azithromycin 1% drop, twice daily for 1wk and then one drop daily for 2wk. The second group got oral doxycycline 100 mg daily for 3wk. At the end of the research, patients' signs and symptoms were compared together. ANOVA, Chi-square and Mann-Whitney tests were used for statistical analysis. RESULTS Topical therapy with azithromycin and oral therapy with doxycycline relieved signs and symptoms after 3wk. There were no significant differences between symptoms healing rate and foreign body sensation healing in these two groups (P>0.05). However, azithromycin drop was more effective in reduction of eye redness and doxycycline was more effective in meibomian glands plugging healing and reducing the corneal staining. CONCLUSION Topical azithromycin could have similar effects as oral doxycycline on posterior blepharitis in improving subjective symptoms. However, doxycycline can reduce objective signs such as ocular surface staining and meibomian gland plugging more than azithromycin. PMID:27500111

  20. Microbial and immunological investigations of chronic non-ulcerative blepharitis and meibomianitis.

    PubMed Central

    Seal, D V; McGill, J I; Jacobs, P; Liakos, G M; Goulding, N J

    1985-01-01

    Concentrations of tear lysozyme, lactoferrin, ceruloplasmin, IgG, and IgA have been measured by enzyme linked immunosorbent assay (ELISA) in patients with chronic non-ulcerative blepharitis and meibomianitis at the same time as the lid and conjunctivae were cultured for bacteria and fungi by a semiquantitative method. A group of normal controls aged 20 to 80 were similarly sampled, when strains of Staphylococcus epidermidis from their eyes and the patients' eyes were biotyped according to Baird-Parker's scheme. 5% of blepharitis cases had increased numbers of Staph. aureus present on the lids, compared with only a scanty growth obtained from 5% of normals. 7% of blepharitis cases had increased numbers of Staph. epidermidis type VI (coagulase-negative, mannitol-fermenting) present compared with a scanty growth obtained from 6% of normals. Isolation rates of other types of Staph. epidermidis did not differ from those in normals; no types were associated with meibomianitis. Tear protein profiles were normal in most patients, and there was no increase in tear IgA or IgG, which is expected with chronic infection. Overall our evidence suggests that in 88% of cases these lid conditions have an inflammatory aetiology not associated with infection. Staphylococcal isolates often found in the eye usually represent a normal commensal rather than pathogenic flora. PMID:4016059

  1. Photochemotherapeutic strategy against Acanthamoeba infections.

    PubMed

    Aqeel, Yousuf; Siddiqui, Ruqaiyyah; Anwar, Ayaz; Shah, Muhammad Raza; Khoja, Shahrukh; Khan, Naveed Ahmed

    2015-01-01

    Acanthamoeba is a protist pathogen that can cause serious human infections, including blinding keratitis and a granulomatous amoebic encephalitis that almost always results in death. The current treatment for these infections includes a mixture of drugs, and even then, a recurrence can occur. Photochemotherapy has shown promise in the treatment of Acanthamoeba infections; however, the selective targeting of pathogenic Acanthamoeba has remained a major concern. The mannose-binding protein is an important adhesin expressed on the surface membranes of pathogenic Acanthamoeba organisms. To specifically target Acanthamoeba, the overall aim of this study was to synthesize a photosensitizing compound (porphyrin) conjugated with mannose and test its efficacy in vitro. The synthesis of mannose-conjugated porphyrin was achieved by mixing benzaldehyde and pyrrole, yielding tetraphenylporphyrin. Tetraphenylporphyrin was then converted into mono-nitrophenylporphyrin by selectively nitrating the para position of the phenyl rings, as confirmed by nuclear magnetic resonance (NMR) spectroscopy. The mono-nitrophenylporphyrin was reduced to mono-aminophenylporphyrin in the presence of tin dichloride and confirmed by a peak at m/z 629. Finally, mono-aminoporphyrin was conjugated with mannose, resulting in the formation of an imine bond. Mannose-conjugated porphyrin was confirmed through spectroscopic analysis and showed that it absorbed light of wavelengths ranging from 425 to 475 nm. To determine the antiacanthamoebic effects of the derived product, amoebae were incubated with mannose-conjugated porphyrin for 1 h and washed 3 times to remove extracellular compound. Next, the amoebae were exposed to light of the appropriate wavelength for 1 h. The results revealed that mannose-conjugated porphyrin produced potent trophicidal effects and blocked excystation. In contrast, Acanthamoeba castellanii incubated with mannose alone and porphyrin alone did not exhibit an antiamoebic effect

  2. Photochemotherapeutic Strategy against Acanthamoeba Infections

    PubMed Central

    Aqeel, Yousuf; Siddiqui, Ruqaiyyah; Anwar, Ayaz; Shah, Muhammad Raza; Khoja, Shahrukh

    2015-01-01

    Acanthamoeba is a protist pathogen that can cause serious human infections, including blinding keratitis and a granulomatous amoebic encephalitis that almost always results in death. The current treatment for these infections includes a mixture of drugs, and even then, a recurrence can occur. Photochemotherapy has shown promise in the treatment of Acanthamoeba infections; however, the selective targeting of pathogenic Acanthamoeba has remained a major concern. The mannose-binding protein is an important adhesin expressed on the surface membranes of pathogenic Acanthamoeba organisms. To specifically target Acanthamoeba, the overall aim of this study was to synthesize a photosensitizing compound (porphyrin) conjugated with mannose and test its efficacy in vitro. The synthesis of mannose-conjugated porphyrin was achieved by mixing benzaldehyde and pyrrole, yielding tetraphenylporphyrin. Tetraphenylporphyrin was then converted into mono-nitrophenylporphyrin by selectively nitrating the para position of the phenyl rings, as confirmed by nuclear magnetic resonance (NMR) spectroscopy. The mono-nitrophenylporphyrin was reduced to mono-aminophenylporphyrin in the presence of tin dichloride and confirmed by a peak at m/z 629. Finally, mono-aminoporphyrin was conjugated with mannose, resulting in the formation of an imine bond. Mannose-conjugated porphyrin was confirmed through spectroscopic analysis and showed that it absorbed light of wavelengths ranging from 425 to 475 nm. To determine the antiacanthamoebic effects of the derived product, amoebae were incubated with mannose-conjugated porphyrin for 1 h and washed 3 times to remove extracellular compound. Next, the amoebae were exposed to light of the appropriate wavelength for 1 h. The results revealed that mannose-conjugated porphyrin produced potent trophicidal effects and blocked excystation. In contrast, Acanthamoeba castellanii incubated with mannose alone and porphyrin alone did not exhibit an antiamoebic effect

  3. Pathogenicity, morphology, and differentiation of Acanthamoeba.

    PubMed

    Khan, N A

    2001-12-01

    Acanthamoeba keratitis is sight threatening corneal infection caused by pathogenic Acanthamoeba. Previous studies have shown the genotypic differences between pathogenic and non-pathogenic species/strains of Acanthamoeba. In this study, we examined the morphological differences between pathogenic and non-pathogenic species/strains using scanning electron microscopy. Pathogenic Acanthamoeba exhibited higher number of acanthopodia (structures associated with the binding of amoeba to the target cells) as compared to non-pathogens. In addition, interactions of amoeba with the corneal epithelial cells were studied. Only pathogenic amoeba exhibited adhesion to epithelial cells. Further results indicated that phagocytosis occurs in the pathogenic amoeba by the formation of amoebastome (characteristic of amoeba phagocyte). This study showed that Acanthamoeba phagocytosis may be both an efficient means of obtaining nutrients for the amoeba and a significant factor in the pathogenesis of Acanthamoeba infections.

  4. Postoperative Corneal and Surgically Induced Astigmatism following Superior Approach Manual Small Incision Cataract Surgery in Patients with Preoperative Against-the-Rule Astigmatism

    PubMed Central

    Sadik, Ahmed Abdul; Mireku, Felix Agyemang; Asiedu, Frank Yeboah; Ablordeppey, Reynolds Kwame

    2016-01-01

    The aim of the study was to report postoperative corneal and surgically induced astigmatism (SIA) in patients with preoperative against-the-rule (ATR) astigmatism who underwent superior approach manual small incision cataract surgery (MSICS). 58 eyes of 58 cataract patients with preoperative ATR astigmatism were involved in this study. All patients had operable cataracts and underwent superior approach MSICS. Keratometric (K) readings were taken prior to surgery and at 12 weeks after surgery. Centroid values of SIA, preoperative astigmatism, and postoperative astigmatism were calculated using Cartesian coordinates based analysis. Wilcoxon signed rank test was used to compute statistical significance between mean preoperative and postoperative corneal astigmatism. Cohen's d was used as effect size measure. Centroid values of 1.42 D × 179, 2.48 D × 0, and 1.07 D × 1 were recorded, respectively, for preoperative astigmatism, postoperative astigmatism, and SIA. Wilcoxon signed rank test indicated that mean ± SD postoperative corneal astigmatism (2.80 ± 1.40 D) was statistically significantly greater than preoperative corneal astigmatism (1.49 ± 1.34 D), Z = −6.263, p < 0.0001. A high Cohen's d of 1.32 was found. Our results suggest statistical and clinically significant greater postoperative corneal astigmatism than preoperative corneal astigmatism for ATR astigmatism cataract patients who underwent superior approach MSICS. PMID:28116142

  5. [Efficacy of physiotherapy and hygienic procedures in treatment of adults and children with chronic blepharitis and dry eye syndrome].

    PubMed

    Prozornaia, L P; Brzhevskiĭ, V V

    2013-01-01

    110 patients aged from 3 to 42 years old were examined to estimate the efficacy of chronic blepharitis treatment: 50 patients with chronic blepharitis and dry eye syndrome (DES), 28 with DES due to computer vision syndrome and 32 with isolated chronic blepharitis. All patients received eyelid massage. If the secretion was too thick and difficult to evacuate from meibomian glands then duct probing was performed. In addition a complex of hygienic procedures was performed using phytoproducts ("Geltec-Medika", Russia): blepharoshampoo, blepharolotion, blepharogel 1 and 2. Moist warm pads (with blepharolotion and calendula extraction) were applied on the eyelids in 25 patients. Massage and probing of meibomian gland ducts and hygienic procedures were showed to be effective in management of clinical signs of chronic blepharitis including coexisting DES. Moist warm pads improve efficacy of background therapy in patients with meibomian gland hypofunction and have no effect in blepharitis with excessive meibomian gland secretion. Eyelid hygiene was showed to be effective in adults and children as well including infants.

  6. Nonlocal conservation laws of the constant astigmatism equation

    NASA Astrophysics Data System (ADS)

    Hlaváč, Adam; Marvan, Michal

    2017-03-01

    For the constant astigmatism equation, we construct a system of nonlocal conservation laws (an abelian covering) closed under the reciprocal transformations. The corresponding potentials are functionally independent modulo a Wronskian type relation.

  7. The Role of Omega-3 Dietary Supplementation in Blepharitis and Meibomian Gland Dysfunction (An AOS Thesis)

    PubMed Central

    Macsai, Marian S.

    2008-01-01

    Purpose Blepharitis and meibomian gland dysfunction (MGD) are common sources of complaints from patients. To evaluate the effect on ocular symptoms, ocular findings, and serum and meibomian gland contents, patients with blepharitis and MGD were prospectively evaluated to determine the effects of dietary supplementation with omega-3 fatty acids. Methods In a prospective randomized placebo-controlled masked trial, patients with simple obstructive MGD and blepharitis, who had discontinued all topical medications and tetracyclines, received oral omega-3 dietary supplementation consisting of two 1000-mg capsules 3 times a day. Patients were examined every 3 months for 1 year with the Ocular Surface Disease Index (OSDI) objective clinical measures, including tear production and stability, ocular surface and meibomian gland health, and biochemical plasma, red blood cell (RBC), and meibum evaluation. Primary outcome measures were change in tear breakup time (TBUT), meibum score, and overall OSDI score at 1 year. Results At 1 year, the omega-3 group had a 36% and 31% reduction in their omega-6 to omega-3 fatty acid ratios in RBCs and plasma, respectively (P = .3), whereas the placebo group demonstrated no change. At 12 months, the omega-3 group had an improvement in TBUT, OSDI score, and meibum score. Changes in meibum content were observed in the omega-3 group (P = .21); the level of meibum saturated fatty acids decreased. Conclusions This trial demonstrated a decrease in the RBC and plasma ratios of omega-6 to omega-3 in patients taking omega-3 dietary supplementation, as compared to controls, and improvements in their overall OSDI score, TBUT, and meibum score. This is the first demonstration of an induced change in the fatty acid saturation content in meibum as a result of dietary supplementation with omega-3 fatty acids. PMID:19277245

  8. A novel gammaherpesvirus in a large flying fox (Pteropus vampyrus) with blepharitis.

    PubMed

    Paige Brock, A; Cortés-Hinojosa, Galaxia; Plummer, Caryn E; Conway, Julia A; Roff, Shannon R; Childress, April L; Wellehan, James F X

    2013-05-01

    A novel gammaherpesvirus was identified in a large flying fox (Pteropus vampyrus) with conjunctivitis, blepharitis, and meibomianitis by nested polymerase chain reaction and sequencing. Polymerase chain reaction amplification and sequencing of 472 base pairs of the DNA-dependent DNA polymerase gene were used to identify a novel herpesvirus. Bayesian and maximum likelihood phylogenetic analyses indicated that the virus is a member of the genus Percavirus in the subfamily Gammaherpesvirinae. Additional research is needed regarding the association of this virus with conjunctivitis and other ocular pathology. This virus may be useful as a biomarker of stress and may be a useful model of virus recrudescence in Pteropus spp.

  9. [New method for treatment of chronic blepharitis associated with dry eye syndrome].

    PubMed

    Safonova, T N; Zabegaĭlo, A O; Fedorov, A A; Lukisheva, O V

    2014-01-01

    A new combined method for treatment of chronic blepharitis associated with dry eye syndrome is developed. A total of 25 patients were enrolled. The treatment implied the use of artificial tears, Restasis, and Blephasteam spectacle frame. The course included 20 procedures on daily basis. The monitoring period was 1 year. A pronounced anti-inflammatory, reparative, and moistening effect as well as stabilization of the precorneal film was achieved in short time. Owing to the combined approach, negative side effects (such as burning, conjunctival irritation, and fluctuating vision) were minimized. Repeated treatment courses maintained the positive therapeutic effect and prolongated the remission.

  10. Three Methods for Correction of Astigmatism during Phacoemulsification

    PubMed Central

    Mohammad-Rabei, Hossein; Mohammad-Rabei, Elham; Espandar, Goldis; Javadi, Mohammad Ali; Jafarinasab, Mohammad Reza; Hashemian, Seyed Javad; Feizi, Sepehr

    2016-01-01

    Purpose: To compare the safety and efficacy of three methods for correcting pre-existing astigmatism during phacoemulsification. Methods: This prospective, comparative, non-randomized study was conducted from March 2010 to January 2011, and included patients with keratometric astigmatism ≥1.25 D undergoing cataract surgery. Astigmatism was corrected using the following approaches: limbal relaxing incisions (LRI) on the steep meridian, extension and suturing of the phaco incision created at the steep meridian (extended-on-axis incision, EOAI), and toric intraocular lens (tIOL) implantation. Keratometric and refractive astigmatism were evaluated 1, 8, and 24 weeks postoperatively. Results: Eighty-three eyes of 72 patients (35 male and 37 female) with mean age of 62.4 ± 14.3 (range, 41-86) years were enrolled. The astigmatism was corrected by using the LRI, EOAI and tIOL implantation methods in 17, 33 and 33 eyes, respectively. Postoperative uncorrected distance visual acuity (UDVA) was significantly improved in all three groups. The difference in postoperative UDVA was not statistically significant among the study groups throughout follow-up except at week 24, when UCVA was significantly better in the tIOL group as compared to the EOAI group (P = 0.024). There is no statistically significant difference of correction index and index of success between three groups at week 24 (P = 0.085 and P = 0.085 respectively). Conclusion: There was no significant difference in astigmatism reduction among the three methods of astigmatism correction during phacoemulsification. Each of these methods can be used at the discretion of the surgeon. PMID:27413496

  11. Successful Surgical Correction of Astigmatism using Customized Ablation Photorefractive Keratectomy

    PubMed Central

    TAHERI, Hakimeh; RAMIN, Shahrokh

    2016-01-01

    The aim of this study was to determine the change in the degree of astigmatism in patients treated with customized ablation photorefractive keratectomy (PRK). This is a cross-sectional study that involved 92 otherwise healthy subjects with regular and irregular astigmatism ≥ 1.25 D (mean age: 39.09 ± 7.72 years; range: 20–59 years). All study subjects were treated with customized ablation PRK using a Technolas 217p Excimer Laser System. Before and 6 months after the surgery, a refraction assessment was conducted for each subject, and the effectiveness of the surgery for correcting astigmatism was evaluated. There was a significant change in astigmatism based on the results of an automated refraction exam of -1.67 ± 1.03 D (P < 0.001), from -2.51 ± 1.45 D preoperatively to -0.87 ± 0.94 D postoperatively. There was also a significant change in subjective refraction of -2.00 ± 1.25 D (P < 0.001), from -2.46 ± 1.52 D preoperatively to -0.46 ± 0.97 D postoperatively. Therefore, our results show that customized ablation PRK is effective for correcting astigmatism ≥ 1.25 D (P < 0.001). PMID:28293648

  12. Acanthamoeba keratitis after photorefractive keratectomy.

    PubMed

    Kaldawy, Roger M; Sutphin, John E; Wagoner, Michael D

    2002-02-01

    A 37-year-old women developed severe suppurative keratitis immediately after having photorefractive keratectomy in her left eye. The keratitis was unresponsive to intensive topical antibiotic agents and topical and systemic steroids. Although the differential diagnosis included nonmicrobial and fungal keratitis, the clinical course and confocal microscopy suggested, and subsequent histopathologic examination confirmed, a diagnosis of Acanthamoeba keratitis. The amebic contamination probably resulted from exposure of the deepithelialized cornea to contaminated freshwater in a northern Wisconsin marsh. This case emphasizes the importance of encouraging patients with epithelial defects and bandage soft contact lenses to avoid exposure to contaminated freshwater until reepithelialization is complete.

  13. Status of astigmatism-corrected Czerny-Turner spectrometers

    NASA Astrophysics Data System (ADS)

    Li, Xinhang; Dong, Keyan; An, Yan; Wang, Zhenye

    2016-10-01

    In order to analysis and design the Czerny-Turner structure spectrometer with the high resolution and high energy reception, various astigmatism methods of the Czerny-Turner structure are reported. According to the location of plane grating, the astigmatism correction methods are divided into two categories, one is the plane grating in divergent illumination, another is the plane grating in parallel illumination. Basing on the different methods, the anastigmatic principle and methods are analyzed, the merits and demerits of the above methods are summarized and evaluated. The theoretical foundation for design of broadband eliminating astigmatism Czerny-Turner spectrometer and the reference value for the further design work are laid by the summary and analyzing in this paper.

  14. Goos-Hänchen and Imbert-Fedorov shifts for astigmatic Gaussian beams

    NASA Astrophysics Data System (ADS)

    Ornigotti, Marco; Aiello, Andrea

    2015-06-01

    In this work we investigate the role of the beam astigmatism in the Goos-Hänchen and Imbert-Fedorov shift. As a case study, we consider a Gaussian beam focused by an astigmatic lens and we calculate explicitly the corrections to the standard formulas for beam shifts due to the astigmatism induced by the lens. Our results show that the different focusing in the longitudinal and transverse direction introduced by an astigmatic lens may enhance the angular part of the shift.

  15. Myopic astigmatism a substitute for accommodation in pseudophakia.

    PubMed

    Huber, C

    1981-12-16

    The power of an intraocular lens can be calculated before surgery to make the eye emmetropic or ametropic. The physiological mechanism of accommodation however, cannot be restored with an inelastic lens. An increased depth of focus in the implanted eye can be predicted through optical principles alone, if the postoperative ametropia of the implanted eye is a simple myopic astigmatism. This increased depth of focus without accommodation was tested in artificial ametropia and found to be used in nature by the seal. To increase the precision of intraocular lens calculation the average change in corneal power induced at surgery is used to predict the postoperative corneal power. By controlled suture release in the postoperative phase, the amount of induced corneal astigmatism is adjusted to obtain a simple myopic astigmatism. Patients with an intraocular lens and a simple myopic astigmatism as a residual ametropia, are spectacle independent most of the time. They need their glasses only for driving or prolonged reading. The methods used to calculate the postoperative cornea, the postoperative anterior chamber depth and the intraocular lens are described with the corresponding calculator programs for the HP 41C calculator. Clinical results and measurements of the depth of focus are shown in a series of 50 successive implant cases.

  16. Correction of low corneal astigmatism in cataract surgery

    PubMed Central

    Leon, Pia; Pastore, Marco Rocco; Zanei, Andrea; Umari, Ingrid; Messai, Meriem; Negro, Corrado; Tognetto, Daniele

    2015-01-01

    AIM To evaluate and compare aspheric toric intraocular lens (IOL) implantation and aspheric monofocal IOL implantation with limbal relaxing incisions (LRI) to manage low corneal astigmatism (1.0-2.0 D) in cataract surgery. METHODS A prospective randomized comparative clinical study was performed. There were randomly recruited 102 eyes (102 patients) with cataracts associated with corneal astigmatism and divided into two groups. The first group received toric IOL implantation and the second one monofocal IOL implantation with peripheral corneal relaxing incisions. Outcomes considered were: visual acuity, postoperative residual astigmatism, endothelial cell count, the need for spectacles, and patient satisfaction. To determine the postoperative toric axis, all patients who underwent the toric IOL implantation were further evaluated using an OPD Scan III (Nidek Co, Japan). Follow-up lasted 6mo. RESULTS The mean uncorrected distance visual acuity (UCVA) and the best corrected visual acuity (BCVA) demonstrated statistically significant improvement after surgery in both groups. At the end of the follow-up the UCVA was statistically better in the patients with toric IOL implants compared to those patients who underwent implantation of monofocal IOL plus LRI. The mean residual refractive astigmatism was of 0.4 D for the toric IOL group and 1.1 D for the LRI group (P<0.01). No difference was observed in the postoperative endothelial cell count between the two groups. CONCLUSION The two surgical procedures demonstrated a significant decrease in refractive astigmatism. Toric IOL implantation was more effective and predictable compared to the limbal relaxing incision. PMID:26309869

  17. Demodex infections in general Polish population, in patients suffering from blepharitis, and among people who work with microscopes.

    PubMed

    Garbacewicz, Aleksandra; Udziela, Monika; Grytner-Ziecina, Barbara; Szaflik, Jacek P; Szaflik, Jerzy

    2010-01-01

    In this study we examined 264 people to estimate the prevalence of Demodex infection. The subjects were divided into 4 groups. First two groups consisted of young people (mean age 22 years) and seniors (mean age 66 years) respectively. The third group included people who work with microscopes (mean age 44 years). Forth group consisted of patients with diagnosed blepharitis (mean age 65 years). From every individual 3-4 lashes were epilated from the eyelids of both eyes and examined under the microscope for Demodex mites. The statistical analysis based on logistic regression was used to estimate the probability of infection. Age was independent variable significant for this model (p < 0.001). Significant relation between age and probability of Demodex infection was showed. The lowest number of infected individuals was observed in the group of young subjects (only 5%), the highest in the blepharitis patients (74% infected). Among microscope users 30% were infected, in the group of seniors 34% were infected.

  18. Laboratory diagnosis of Acanthamoeba keratitis in Hungary.

    PubMed

    Orosz, Erika; Farkas, Ágnes; Kucsera, István

    2016-09-01

    Acanthamoeba species are free-living amebae that can be found in almost every range of environments. Within this genus, numerous species are recognized as human pathogens, potentially causing Acanthamoeba keratitis (AK). AK is a corneal disease that is predominantly associated with contact lens use, the epidemiology of which is related to the specific genotype of Acanthamoeba. This study reports seven (7/16; 43.75%) positive cases. Detection of Acanthamoeba in corneal scrapings is based on cultivation and polymerase chain reaction (PCR) combined with the molecular taxonomic identification method. By PCR, seven samples were positive; cultivation was successful for five samples, probably because of the low quantity of samples. Genotype identification was carried out with a real-time fluorescence resonance energy transfer PCR assay based on sequence analysis of the 18S rRNA gene, and sensitivity and specificity were evaluated in comparison with traditional parasitological techniques. All seven detected Acanthamoeba strains belonged to the T4 genotype, the main AK-related genotype worldwide. These results confirmed the importance of a complete diagnostic protocol, including a PCR assay, for the clinical diagnosis of AK from human samples. Genotyping allowed the identification of all isolates in the T4 group, thus demonstrating the prevalence of this genotype in Hungary.

  19. First cases of Acanthamoeba keratitis in Slovakia.

    PubMed

    Ondriska, Frantisek; Mrva, Martin; Lichvár, Martin; Ziak, Peter; Murgasová, Zuzana; Nohýnková, Eva

    2004-01-01

    We present the case report of the first identification of Acanthamoeba as a causative agent of keratitis in the Slovak Republic. For the first time, Acanthamoeba sp. Group III was isolated from a 53-year-old patient with keratitis, which was manifested after an injury of the right eye. A delayed visit to a physician as well as a late diagnosis of the illness led to the advanced stage of eye disease. As the treatment with itraconazol and cornea transplantation showed no result, enucleation of the eye was decided. Acanthamoeba ludgunensis was also the causative agent of keratitis in a 39-year-old patient wearing contact lenses. His complaints occurred a month after bathing in a thermal swimming pool. The symptoms presented in the left eye were those of herpetic keratitis, and led to a cloudy cornea with circular infliltrate and poor vision. A prompt clinical and laboratory diagnosis, along with treatment with propamidine-isetionate resulted in a significant improvement of the eye condition. Contact lenses were probably related to another case of Acanthamoeba keratitis. The patient, a 15-year-old girl, kept wearing contact lenses during bathing in various swimming pools and in the sea; her contact lenses were also regularly washed under tap water. Due to the fact that cysts of Acanthamoeba sp. group II were found in the contact lens solution, this is presumed to be the source of the eye infection.

  20. A case of trauma related Acanthamoeba keratitis.

    PubMed

    Kamel, A G M; Faridah, H; Yusof, S; Norazah, A; Nakisah, M A

    2004-12-01

    Acanthamoeba is an uncommon cause of keratitis but one of the most severe because of the prolonged and painful course of the disease and poor visual outcome. Although contact lens use is the principal risk factor, about 10% of cases occur following trauma and exposure to contaminated soil or water. Two cases of Acanthamoeba keratitis involving women contact lens wearers have previously been reported in Malaysia but this is the first time, a non contact lens related Acanthamoeba keratitis is reported. The case involved a 28 year old Indonesian male construction worker who had a trauma of the right eye during work. His eye was struck by sand and dust particles after which he quickly washed with water from an open tank at the construction site. He experienced pain, redness, glaring and blurring of vision of the right eye three days later. The diagnosis was missed at initial presentation but culture of the corneal scraping had proven Acanthamoeba as the aetiological agent. The history and clinical findings of this trauma related Acanthamoeba keratitis are briefly discussed.

  1. Correlation Between Stereoacuity and Experimentally Induced Graded Monocular and Binocular Astigmatism

    PubMed Central

    Puthran, Neelam; Gagal, Bhavna

    2016-01-01

    Introduction Stereopsis, the highest grade of binocular single vision, is affected by various factors, such as mis-alignment of visual axes, refractive errors especially anisometropia and astigmatism, both of which may result in amblyopia. There are very few studies in literature regarding the relationship between stereoacuity and refractive errors, especially astigmatism. Aim The present study was conducted to determine the correlation between stereoacuity and experimentally induced graded astigmatism in emmetropes. Materials and Methods A randomized study was conducted on 2000 individuals of either gender, between the ages of 8-35 years, at tertiary care centre attached to a medical college during the period of August 2012 to August 2014, All subjects were emmetropic with normal binocular single vision. Participants were randomly divided into four groups of 500 individuals each. Two groups were subjected to induced myopic astigmatism, either uni-ocularly or binocularly, using + 1.0 D and + 2.0 D cylinders at varying axes i.e., 450, 900 and 1800. Similarily, the remaining two groups were subjected to induced hypermetropic astigmatism, using - 1.0 D and - 2.0D cylinders at varying axes i.e. 450, 900 and 1800. Near stereoacuity was determined by the Titmus Fly Stereo Test, both before and after induction of astigmatism. Statistical analysis was done using paired t-test and ANOVA. Results The mean stereoacuity in emmetropes was 28.81±4.97 seconds of arc. There was a decrease in stereoacuity with increase in dioptric power of astigmatism (p<0.001). Oblique astigmatism reduced the stereoacuity maximally, while stereoacuity was least affected at 180o axis. Hypermetropic astigmatism caused more deterioration in stereoacuity than myopic astigmatism. A gross reduction in stereoacuity was noted in induced monocular astigmatism as against binocular astigmatism. Conclusion This study suggests that stereoacuity is significantly affected by even minor degrees of monocular or

  2. Effect of Mitomycin C on Myopic versus Astigmatic Photorefractive Keratectomy

    PubMed Central

    Fawzy, Samah M.

    2017-01-01

    Purpose. Long-term mitomycin C (MMC) effects on photorefractive keratectomy (PRK) were compared in simple myopic and astigmatic patients. Methods. In this observational cohort study, subjects were selected based on preoperative and postoperative data collected from medical records; they were divided into simple myopia with/without MMC and myopic astigmatism with/without MMC groups. Haze, uncorrected visual acuity (UCVA), best-corrected visual acuity (BCVA), subjective refraction, and K-reading were evaluated at 1-, 3-, 6-, and 12-month follow-ups. Results. One hundred fifty-nine eyes of 80 subjects (34 women and 46 men; mean age, 26.81 ± 7.74 years; range, 18–53 years; spherical powers, −0.50 to −8.00 DS; and cylindrical powers, −0.25 to −5.00 DC) were enrolled. One year postoperatively, the simple myopia with/without MMC groups showed no difference in UCVA (P = 0.187), BCVA (P = 0.163), or spherical equivalent (P = 0.163) and a significant difference (P = 0.0495) in K-reading; the haze formation difference was nonsignificant (P = 0.056). Astigmatic groups with/without MMC showed a significant difference in K-reading (P < 0.0001). MMC groups had less haze formation (P < 0.0001). Conclusion. PRK with intraoperative MMC application showed excellent visual outcomes. MMC's effect on astigmatic patients was significantly better with acceptable safety and minimal side effects. PMID:28392938

  3. [Results of refractive surgery in hyperopic and combined astigmatism].

    PubMed

    Vlaicu, Valeria

    2013-01-01

    The refractive surgery includes a lot of procedures for changing the refraction of the eye to obtain a better visual acuity with no glasses or contact lenses. LASIK is the most commonly performed laser refractive surgery today. The goal is to present the postoperative evolution of the refraction and visual acuity after LASIK for Mixed and Hyperopic Astigmatism. The results show that LASIK is safe and predictible if we have well performed interventions and well-selected patients.

  4. Metrology of complex astigmatic surfaces for astronomical optics

    NASA Astrophysics Data System (ADS)

    Rolt, Stephen; Kirby, Andrew K.; Robertson, David J.

    2010-07-01

    This paper will focus on the metrology of multiple complex surfaces that are to be integrated into the KBand Multi- Object Spectrograph (KMOS). KMOS is a multi-field astronomical spectrograph designed for integration with the 8.2m diameter European Southern Observatory Very Large Telescope (VLT). There are 1080 separate optical surfaces in the design, many of them complex freeform surfaces. Optical surfaces were manufactured in aluminium by precision freeform diamond machining. This flexible technique allows the fabrication of extremely complex surfaces with an accuracy of better than 15 nm RMS over a 20 mm aperture, giving the designer great freedom in generating powerful and unorthodox designs. However, the complexity of these freeform surfaces poses a challenge to their accurate characterisation. This paper will discuss in detail the metrology of a specific freeform component in the instrument. The form of these complex astigmatic surfaces was measured using spherical wavefronts by adapting a tilted Twyman-Green Interferometer arrangement. There are eight separate designs for this type of component, each with a different orientation and magnitude of astigmatism. Careful mechanical fixturing is essential to align the astigmatic axis to the test set up. The impact of mechanical tolerances on measurement uncertainty will be discussed in detail.

  5. Development of the fast astigmatic auto-focus microscope system

    NASA Astrophysics Data System (ADS)

    Hsu, Wei-Yao; Lee, Chien-Shing; Chen, Po-Jui; Chen, Nien-Tsu; Chen, Fong-Zhi; Yu, Zong-Ru; Kuo, Ching-Hsiang; Hwang, Chi-Hung

    2009-04-01

    In this paper, a fast auto-focus microscope system was developed based on the astigmatic method. A collimated infrared laser beam was employed in the infinite-corrected microscope optical axis by the beam splitter and reflected by the sample surface. By embedding an astigmatic lens in the system, the reflected laser beam has different focal lengths in the sagittal and tangential planes. As the microscope's relative distance varies, the reflected laser beam shape also varies and can be detected by an embedded four-quadrant photodiode, i.e., the focus error signal (FES) can be found. Then, a fast auto-focus system can be realized by converting the FES to the microscope's defocus distance. We designed an astigmatic auto-focus system for a 20× objective lens with a ±50 µm working range, and this system could also be used for 10× and 5× objectives with ±200 µm and ±800 µm working ranges, respectively.

  6. Molecular and physiological evaluation of subtropical environmental isolates of Acanthamoeba spp., causal agent of Acanthamoeba keratitis.

    PubMed

    Booton, Gregory C; Rogerson, Andrew; Bonilla, Tonya D; Seal, David V; Kelly, Daryl J; Beattie, Tara K; Tomlinson, Alan; Lares-Villa, Fernando; Fuerst, Paul A; Byers, Thomas J

    2004-01-01

    Previous molecular examination of Acanthamoeba spp. has resulted in the determination of distinct genotypes in this genus (designated T1-T12, T14). Genotype T4 has been responsible for the majority of cases of Acanthamoeba keratitis. Here we examine the relative abundance of environmental T4 isolates on beaches and ask whether they have temperature and salinity tolerances that could enhance pathogenicity. Twenty-four Acanthamoeba strains were isolated from beach sand (n = 20), soil (n = 3), and tap water (n = 1) in south Florida. Phylogenetic analysis identified 19 of 24 isolates as T4, the Acanthamoeba keratitis-associated genotype. The remaining isolates were genotype T5 (4) and T11 (1). Nearly all beach isolates were genotype T4, whereas the tap water and soil isolates were mostly T5. All amoebae grew at 0, 1.0, and 2.0% salt and 19 of 20 beach isolates also grew at 3.2%. No soil or tap-water acanthamoebae reproduced at 3.2%. All isolates grew at 37 degrees C and two (T5) at 42 degrees C. Little correlation existed between beach location, salt-tolerance, and genetic relatedness. Overall, the large majority of environmental isolates obtained were genotype T4, suggesting it may be the most common genotype in this environment and could be a potential source of Acanthamoeba keratitis infections.

  7. Autophagy Inhibitors as a Potential Antiamoebic Treatment for Acanthamoeba Keratitis

    PubMed Central

    Moon, Eun-Kyung; Kim, So-Hee; Hong, Yeonchul; Chung, Dong-Il; Goo, Youn-Kyoung

    2015-01-01

    Acanthamoeba cysts are resistant to extreme physical and chemical conditions. Autophagy is an essential pathway for encystation of Acanthamoeba cells. To evaluate the possibility of an autophagic Acanthamoeba encystation mechanism, we evaluated autophagy inhibitors, such as 3-methyladenine (3MA), LY294002, wortmannin, bafilomycin A, and chloroquine. Among these autophagy inhibitors, the use of 3MA and chloroquine showed a significant reduction in the encystation ratio in Acanthamoeba cells. Wortmannin also inhibited the formation of mature cysts, while LY294002 and bafilomycin A did not affect the encystation of Acanthamoeba cells. Transmission electron microscopy revealed that 3MA and wortmannin inhibited autophagy formation and that chloroquine interfered with the formation of autolysosomes. Inhibition of autophagy or autolysosome formation resulted in a significant block in the encystation in Acanthamoeba cells. Clinical treatment with 0.02% polyhexamethylene biguanide (PHMB) showed high cytopathic effects on Acanthamoeba trophozoites and cysts; however, it also revealed high cytopathic effects on human corneal epithelial cells. In this study, we investigated effects of the combination of a low (0.00125%) concentration of PHMB with each of the autophagy inhibitors 3MA, wortmannin, and chloroquine on Acanthamoeba and human corneal epithelial cells. These new combination treatments showed low cytopathic effects on human corneal cells and high cytopathic effects on Acanthamoeba cells. Taken together, these results provide fundamental information for optimizing the treatment of Acanthamoeba keratitis. PMID:25896709

  8. Autophagy inhibitors as a potential antiamoebic treatment for Acanthamoeba keratitis.

    PubMed

    Moon, Eun-Kyung; Kim, So-Hee; Hong, Yeonchul; Chung, Dong-Il; Goo, Youn-Kyoung; Kong, Hyun-Hee

    2015-07-01

    Acanthamoeba cysts are resistant to extreme physical and chemical conditions. Autophagy is an essential pathway for encystation of Acanthamoeba cells. To evaluate the possibility of an autophagic Acanthamoeba encystation mechanism, we evaluated autophagy inhibitors, such as 3-methyladenine (3MA), LY294002, wortmannin, bafilomycin A, and chloroquine. Among these autophagy inhibitors, the use of 3MA and chloroquine showed a significant reduction in the encystation ratio in Acanthamoeba cells. Wortmannin also inhibited the formation of mature cysts, while LY294002 and bafilomycin A did not affect the encystation of Acanthamoeba cells. Transmission electron microscopy revealed that 3MA and wortmannin inhibited autophagy formation and that chloroquine interfered with the formation of autolysosomes. Inhibition of autophagy or autolysosome formation resulted in a significant block in the encystation in Acanthamoeba cells. Clinical treatment with 0.02% polyhexamethylene biguanide (PHMB) showed high cytopathic effects on Acanthamoeba trophozoites and cysts; however, it also revealed high cytopathic effects on human corneal epithelial cells. In this study, we investigated effects of the combination of a low (0.00125%) concentration of PHMB with each of the autophagy inhibitors 3MA, wortmannin, and chloroquine on Acanthamoeba and human corneal epithelial cells. These new combination treatments showed low cytopathic effects on human corneal cells and high cytopathic effects on Acanthamoeba cells. Taken together, these results provide fundamental information for optimizing the treatment of Acanthamoeba keratitis.

  9. Success rates in the correction of astigmatism with toric and spherical soft contact lens fittings

    PubMed Central

    Kurna, Sevda Aydin; Şengör, Tomris; Ün, Murat; Aki, Suat

    2010-01-01

    Objectives: To evaluate success rates in the correction of astigmatism with toric and spherical soft contact lens fitting. Methods: 30 patients with soft toric lenses having more than 1.25 D of corneal astigmatism (25 eyes; Group A) or having 0.75–1.25 D of corneal astigmatism (22 eyes; Group B) and 30 patients with soft spheric lenses having 0.75–1.25 D of corneal astigmatism (28 eyes; Group C) or less than 0.75 D of corneal astigmatism (23 eyes; Group D) were included in the study. Corrected and uncorrected monocular visual acuity measurement with logMAR, biomicroscopic properties, autorefractometry and corneal topography were performed for all patients immediately before and at least 20 minutes after the application of contact lenses. Success of contact lens fitting was evaluated by three parameters: astigmatic neutralization, visual success, and retinal deviation. Results: After soft toric lens application, spheric dioptres, cylindric and keratometric astigmatism, and retinal deviation decreased significantly in Groups A and B (P < 0.05). In Group C, spheric dioptres and retinal deviation decreased (P < 0.05), while cylindric and keratometric astigmatism did not change significantly (P > 0.05). In Group D, spheric dioptres, retinal deviation, and cylindric astigmatism decreased (P < 0.05). Keratometric astigmatism did not change significantly (P > 0.05) and astigmatic neutralization even increased. Conclusions: Visual acuity and residual spherical equivalent refraction remained between tolerable limits with the use of toric and spheric contact lenses. Spherical lenses failed to mask corneal toricity during topography, while toric lenses caused central neutralization and decrease in corneal cylinder in low and moderate astigmatic eyes. PMID:20856589

  10. Correction of high amounts of astigmatism through orthokeratology. A case report

    PubMed Central

    Baertschi, Michael; Wyss, Michael

    2011-01-01

    The purpose of this case report is to introduce a method for a successful treatment of high astigmatism with a new orthokeratology design, called FOKX (Falco Kontaktlinsen, Switzerland). This novel toric orthokeratology contact lens design, the fitting approach and the performance of FOKX lenses will be illustrated in the form of a case report. Correcting astigmatism with orthokeratology offers a new perspective for all patients suffering astigmatism.

  11. Propagation of electromagnetic multi-Gaussian Schell-model beams with astigmatic aberration in turbulent ocean.

    PubMed

    Lu, Chuanyi; Zhao, Daomu

    2016-10-10

    An analytical expression for the elements of the cross-spectral density matrix of electromagnetic multi-Gaussian Schell-model beams with astigmatic aberration propagating through the turbulent ocean is derived. We investigate the statistical characteristics of the beams on propagation in great detail. It is found that, due to astigmatism, the circular symmetry of such beams suffers a certain degree of damage in the near field. In addition, astigmatism also destroys the attractive far-field flat profiles in free space. Meanwhile, we also show that astigmatism and oceanic turbulence both have certain effects on the spectral degree of polarization.

  12. Effect of astigmatism on states of polarization of aberrant stochastic electromagnetic beams in turbulent atmosphere.

    PubMed

    Li, Jia; Chen, Yanru; Zhao, Qi; Zhou, Muchun

    2009-10-01

    The effect of astigmatism on states of polarization of aberrant stochastic electromagnetic beams in turbulent atmosphere is investigated. Using the Gaussian-Schell model source with astigmatism, the analytical formula for the degree of polarization, the orientation angle, and the degree of polarization ellipse are derived. Analytical results show that different strengths of astigmatism have different effects on states of polarization on propagation. It is also shown that when the astigmatic coefficient of sources is large enough, states of polarization are hardly affected by atmospheric turbulence and the free-space diffraction phenomenon. The sufficient conditions for propagating with invariant polarization are derived and discussed.

  13. Twenty Years of Acanthamoeba Diagnostics in Austria

    PubMed Central

    Walochnik, Julia; Scheikl, Ute; Haller-Schober, Eva-Maria

    2015-01-01

    Acanthamoebae are the causative agents of an often seriously progressing keratitis (AK) occurring predominantly in contact lens wearers and can cause several disseminating infections potentially resulting in granulomatous amoebic encephalitis (GAE) in the immunocompromised host. Our institution is the Austrian reference laboratory for Acanthamoeba diagnostics and the aim of this study was to give an overview of proven cases of Acanthamoeba infections in Austria during the past 20 yr. All samples of patients with suspected AK or GAE were screened for Acanthamoeba spp. by culture and/or PCR and the detected amoebae were genotyped. Altogether, 154 cases of AK and three cases of GAE were diagnosed. Age of the AK patients ranged from 8 to 82 yr (mean 37.8) and 58% of the patients were female. Approximately 89% of the AK patients were contact lens wearers, almost all cases were unilateral and 19% of the patients required a keratoplasty. Age of the GAE patients ranged from 2 to 25 yr (mean 14.7), all were HIV-negative, but two were severely immunosuppressed at the time of diagnosis. The predominant genotype in the AK cases was T4, other genotypes found were T3, T5, T6, T10 and T11. The three GAE cases involved genotypes T2, T4 and T5. PMID:25047131

  14. Twenty years of acanthamoeba diagnostics in Austria.

    PubMed

    Walochnik, Julia; Scheikl, Ute; Haller-Schober, Eva-Maria

    2015-01-01

    Acanthamoebae are the causative agents of an often seriously progressing keratitis (AK) occurring predominantly in contact lens wearers and can cause several disseminating infections potentially resulting in granulomatous amoebic encephalitis (GAE) in the immunocompromised host. Our institution is the Austrian reference laboratory for Acanthamoeba diagnostics and the aim of this study was to give an overview of proven cases of Acanthamoeba infections in Austria during the past 20 yr. All samples of patients with suspected AK or GAE were screened for Acanthamoeba spp. by culture and/or PCR and the detected amoebae were genotyped. Altogether, 154 cases of AK and three cases of GAE were diagnosed. Age of the AK patients ranged from 8 to 82 yr (mean 37.8) and 58% of the patients were female. Approximately 89% of the AK patients were contact lens wearers, almost all cases were unilateral and 19% of the patients required a keratoplasty. Age of the GAE patients ranged from 2 to 25 yr (mean 14.7), all were HIV-negative, but two were severely immunosuppressed at the time of diagnosis. The predominant genotype in the AK cases was T4, other genotypes found were T3, T5, T6, T10 and T11. The three GAE cases involved genotypes T2, T4 and T5.

  15. Medical interventions for acanthamoeba keratitis

    PubMed Central

    Alkharashi, Majed; Lindsley, Kristina; Law, Hua Andrew; Sikder, Shameema

    2016-01-01

    Background Acanthamoeba are microscopic, free-living, single-celled organisms which can infect the eye and lead to Acanthamoeba keratitis (AK). AK can result in loss of vision in the infected eye or loss of eye itself; however, there are no formal guidelines or standards of care for the treatment of AK. Objectives To evaluate the relative effectiveness and safety of medical therapy for the treatment of AK. Search methods We searched CENTRAL (which contains the Cochrane Eyes and Vision Group Trials Register) (2015, Issue 1), Ovid MEDLINE, Ovid MEDLINE In-Process and Other Non-Indexed Citations, Ovid MEDLINE Daily, Ovid OLDMEDLINE (January 1946 to January 2015), EMBASE (January 1980 to January 2015), PubMed (1948 to January 2015), Latin American and Caribbean Health Sciences Literature Database (LILACS) (1982 to January 2015), the metaRegister of Controlled Trials (mRCT) (www.controlled-trials.com), ClinicalTrials.gov (www.clinicaltrials.gov) and the World Health Organization (WHO) International Clinical Trials Registry Platform (ICTRP) (www.who.int/ictrp/search/en). We did not use any date or language restrictions in the electronic search for trials. We last searched the electronic databases on 9 January 2015. Selection criteria We included randomized controlled trials (RCTs) of medical therapy for AK, regardless of the participants' age, sex, or etiology of disease. We included studies that compared either anti-amoeba therapy (drugs used alone or in combination with other medical therapies) with no anti-amoeba therapy or one anti-amoeba therapy with another anti-amoeba therapy. Data collection and analysis Two authors independently screened search results and full-text reports, assessed risk of bias, and abstracted data. We used standard methodological procedures as set forth by the Cochrane Collaboration. Main results We included one RCT (56 eyes of 55 participants) in this review. The study compared two types of topical biguanides for the treatment of AK

  16. A clinical Acanthamoeba isolate harboring two distinct bacterial endosymbionts.

    PubMed

    Müller, Anneliese; Walochnik, Julia; Wagner, Martin; Schmitz-Esser, Stephan

    2016-10-01

    Acanthamoebae feed on bacteria but are also frequent hosts of bacterial symbionts. Here, we describe the stable co-occurrence of two symbionts, one affiliated to the genus Parachlamydia and the other to the candidate genus Paracaedibacter (Alphaproteobacteria), within a clinical isolate of Acanthamoeba hatchetti genotype T4. We performed fluorescence in situ hybridization (FISH) and transmission electron microscopy (TEM) to describe this symbiosis. Our study adds to other reports of simultaneous co-occurrence of two symbionts within one Acanthamoeba cell.

  17. Acanthamoeba: biology and increasing importance in human health.

    PubMed

    Khan, Naveed Ahmed

    2006-07-01

    Acanthamoeba is an opportunistic protozoan that is widely distributed in the environment and is well recognized to produce serious human infections, including a blinding keratitis and a fatal encephalitis. This review presents our current understanding of the burden of Acanthamoeba infections on human health, their pathogenesis and pathophysiology, and molecular mechanisms associated with the disease, as well as virulence traits of Acanthamoeba that may be targets for therapeutic interventions and/or the development of preventative measures.

  18. Area 51: How do Acanthamoeba invade the central nervous system?

    PubMed

    Siddiqui, Ruqaiyyah; Emes, Richard; Elsheikha, Hany; Khan, Naveed Ahmed

    2011-05-01

    Acanthamoeba granulomatous encephalitis generally develops as a result of haematogenous spread, but it is unclear how circulating amoebae enter the central nervous system (CNS) and cause inflammation. At present, the mechanisms which Acanthamoeba use to invade this incredibly well-protected area of the CNS and produce infection are not well understood. In this paper, we propose two key virulence factors: mannose-binding protein and extracellular serine proteases as key players in Acanthamoeba traversal of the blood-brain barrier leading to neuronal injury. Both molecules should provide excellent opportunities as potential targets in the rational development of therapeutic interventions against Acanthamoeba encephalitis.

  19. Phylogenetic evidence for a new genotype of Acanthamoeba (Amoebozoa, Acanthamoebida).

    PubMed

    Corsaro, Daniele; Venditti, Danielle

    2010-06-01

    Acanthamoeba are widespread free-living amoebae, able to cause infection in animals, with keratitis and granulomatous encephalitis as major diseases in humans. Recent developments in the subgenus classification are based on the determination of the nucleotide sequence of the 18S rDNA. By this mean, Acanthamoeba have been clustered into 15 sequence types or genotypes, called T1 to T15. In this study, we analysed near full 18S rDNA of an Acanthamoeba recovered from an environmental sample and various unidentified Acanthamoeba sequences retrieved from GenBank. We provided phylogenetic evidence for a new genotype, which we proposed to name T16.

  20. Novel Parachlamydia acanthamoebae quantification method based on coculture with amoebae.

    PubMed

    Matsuo, Junji; Hayashi, Yasuhiro; Nakamura, Shinji; Sato, Marie; Mizutani, Yoshihiko; Asaka, Masahiro; Yamaguchi, Hiroyuki

    2008-10-01

    Parachlamydia acanthamoebae, belonging to the order Chlamydiales, is an obligately intracellular bacterium that infects free-living amoebae and is a potential human pathogen. However, no method exists to accurately quantify viable bacterial numbers. We present a novel quantification method for P. acanthamoebae based on coculture with amoebae. P. acanthamoebae was cultured either with Acanthamoeba spp. or with mammalian epithelial HEp-2 or Vero cells. The infection rate of P. acanthamoebae (amoeba-infectious dose [AID]) was determined by DAPI (4',6-diamidino-2-phenylindole) staining and was confirmed by fluorescent in situ hybridization. AIDs were plotted as logistic sigmoid dilution curves, and P. acanthamoebae numbers, defined as amoeba-infectious units (AIU), were calculated. During culture, amoeba numbers and viabilities did not change, and amoebae did not change from trophozoites to cysts. Eight amoeba strains showed similar levels of P. acanthamoebae growth, and bacterial numbers reached ca. 1,000-fold (10(9) AIU preculture) after 4 days. In contrast, no increase was observed for P. acanthamoebae in either mammalian cell line. However, aberrant structures in epithelial cells, implying possible persistent infection, were seen by transmission electron microscopy. Thus, our method could monitor numbers of P. acanthamoebae bacteria in host cells and may be useful for understanding chlamydiae present in the natural environment as human pathogens.

  1. Is there evidence of sexual reproduction (meiosis) in Acanthamoeba?

    PubMed

    Khan, Naveed A; Siddiqui, Ruqaiyyah

    2015-06-01

    Evolution of independently breeding species into males and females (gametes) has remained a puzzle. Given the significant advantages of sexual reproduction over asexual reproduction as a long-term species survival strategy; here, we pose the question whether there is some form of meiosis in Acanthamoeba species, which represents our ancient lineage. The recently available Acanthamoeba genome revealed several genes implicated in meiosis in sexual eukaryotes such as Spo11, Mre11, Rad50, Rad51, Rad52, Mnd1, Dmc1, Msh, and Mlh, suggesting that Acanthamoeba is capable of some form of meiosis, inferring the presence of sexual reproduction in Acanthamoeba, and that meiosis evolved early in eukaryotic evolution.

  2. Propagation of the Lissajous singularity dipole through an astigmatic lens

    NASA Astrophysics Data System (ADS)

    Chen, Haitao; Gao, Zenghui; Zou, Xuefang; Huang, Weigang; Wang, Fanhou

    2013-11-01

    The propagation was investigated of a pair of Lissajous singularities of opposite singularity index called the Lissajous singularity dipole (LSD) through an astigmatic lens. It is shown that after passing through the lens the positions of the singularities are inverted and more than two LSDs occur. Changes in the degree of polarization of the LSDs as well as in the shape of the Lissajous figures also occur. In addition, Lissajous singularities may take place at the focal plane, and a single Lissajous singularity may appear and vanish under certain conditions. The results are compared with those of previous work.

  3. Relaxing incision for control of postoperative astigmatism following keratoplasty.

    PubMed

    Troutman, R C; Swinger, C

    1980-02-01

    A new technique has been presented which provides a second surgical approach to correct excessive residual astigmatism following keratoplasty. The relaxing incision procedure has advantages over wedge resection in that it can be performed at the slit lamp, it gives no initial overcorrection, and it has a much shorter postoperative course while giving rapid results without suturing. Since this technique does not appear to produce significant hyperopia and may produce a tendency toward myopia, the corneal surgeon would thus have alternative techniques from which to choose, depending, in part, on the spherical component of the refraction.

  4. Ocular surface discomfort and Demodex: effect of tea tree oil eyelid scrub in Demodex blepharitis.

    PubMed

    Koo, Hyun; Kim, Tae Hyung; Kim, Kyoung Woo; Wee, Sung Wook; Chun, Yeoun Sook; Kim, Jae Chan

    2012-12-01

    The purpose of this study was to evaluate the relation between ocular discomfort and ocular Demodex infestation, and therapeutic effects of tea tree oil (TTO) in Demodex blepharitis patients. Three hundred and thirty-five patients with ocular discomfort were evaluated for ocular Demodex infestation and subjective symptoms with ocular surface discomfort index (OSDI) score. Among them, Demodex-infested patients were randomized to receive either eyelid scrubbing with TTO (TTO group,106 patients) or without TTO (Control group, 54 patients) for 1 month. Demodex were found in 84% of patients with ocular discomfort. The number of Demodex was significantly correlated with age (P = 0.04) and OSDI score (P = 0.024). After eyelid scrub treatment, Demodex count was reduced from 4.0 ± 2.5 to 3.2 ± 2.3 in the TTO group (P = 0.004) and from 4.3 ± 2.7 to 4.2 ± 2.5 in the control group (P = 0.27). Also, OSDI score was reduced from 34.5 ± 10.7 to 24.1 ± 11.9 in the TTO group (P = 0.001) and from 35.3 ± 11.6 to 27.5 ± 12.8 in the control group (P = 0.04). In conclusion, Demodex number showed a significant positive correlation with age and subjective ocular discomfort. The tea tree oil eyelid scrub treatment is effective for eliminating ocular Demodex and improving subjective ocular symptoms.

  5. Ocular Surface Discomfort and Demodex: Effect of Tea Tree Oil Eyelid Scrub in Demodex Blepharitis

    PubMed Central

    Koo, Hyun; Kim, Tae Hyung; Kim, Kyoung Woo; Wee, Sung Wook; Chun, Yeoun Sook

    2012-01-01

    The purpose of this study was to evaluate the relation between ocular discomfort and ocular Demodex infestation, and therapeutic effects of tea tree oil (TTO) in Demodex blepharitis patients. Three hundred and thirty-five patients with ocular discomfort were evaluated for ocular Demodex infestation and subjective symptoms with ocular surface discomfort index (OSDI) score. Among them, Demodex-infested patients were randomized to receive either eyelid scrubbing with TTO (TTO group,106 patients) or without TTO (Control group, 54 patients) for 1 month. Demodex were found in 84% of patients with ocular discomfort. The number of Demodex was significantly correlated with age (P = 0.04) and OSDI score (P = 0.024). After eyelid scrub treatment, Demodex count was reduced from 4.0 ± 2.5 to 3.2 ± 2.3 in the TTO group (P = 0.004) and from 4.3 ± 2.7 to 4.2 ± 2.5 in the control group (P = 0.27). Also, OSDI score was reduced from 34.5 ± 10.7 to 24.1 ± 11.9 in the TTO group (P = 0.001) and from 35.3 ± 11.6 to 27.5 ± 12.8 in the control group (P = 0.04). In conclusion, Demodex number showed a significant positive correlation with age and subjective ocular discomfort. The tea tree oil eyelid scrub treatment is effective for eliminating ocular Demodex and improving subjective ocular symptoms. PMID:23255861

  6. Acanthamoeba keratitis: improving the Scottish diagnostic service for the rapid molecular detection of Acanthamoeba species.

    PubMed

    Alexander, Claire Low; Coyne, Michael; Jones, Brian; Anijeet, Deepa

    2015-07-01

    Acanthamoeba species are responsible for causing the potentially sight-threatening condition, Acanthamoeba keratitis, which is commonly associated with contact lens use. In this report, we highlight the challenges faced using conventional laboratory identification methods to identify this often under-reported pathogen, and discuss the reasons for introducing the first national service in Scotland for the rapid and sensitive molecular identification of Acanthamoeba species. By comparing culture and molecular testing data from a total of 63 patients (n = 80 samples) throughout Scotland presenting with ocular eye disease, we describe the improvement in detection rates where an additional four positive cases were identified using a molecular assay versus culture. The testing of a further ten patients by confocal imaging is also presented. This report emphasizes the importance of continuing to improve clinical laboratory services to ensure a prompt, correct diagnosis and better prognosis, in addition to raising awareness of this potentially debilitating opportunistic pathogen.

  7. The Development of Drugs against Acanthamoeba Infections.

    PubMed

    Siddiqui, Ruqaiyyah; Aqeel, Yousuf; Khan, Naveed Ahmed

    2016-11-01

    For the past several decades, there has been little improvement in the morbidity and mortality associated with Acanthamoeba keratitis and Acanthamoeba encephalitis, respectively. The discovery of a plethora of antiacanthamoebic compounds has not yielded effective marketed chemotherapeutics. The rate of development of novel antiacanthamoebic chemotherapies of translational value and the lack of interest of the pharmaceutical industry in developing such chemotherapies have been disappointing. On the other hand, the market for contact lenses/contact lens disinfectants is a multi-billion-dollar industry and has been successful and profitable. A better understanding of drugs, their targets, and mechanisms of action will facilitate the development of more-effective chemotherapies. Here, we review the progress toward phenotypic drug discovery, emphasizing the shortcomings of useable therapies.

  8. A type-1 metacaspase from Acanthamoeba castellanii.

    PubMed

    Trzyna, Wendy C; Legras, Xavier D; Cordingley, John S

    2008-01-01

    The complete sequence of a type-1 metacaspase from Acanthamoeba castellanii is reported comprising 478 amino acids. The metacaspase was recovered from an expression library using sera specific for membrane components implicated in stimulating encystation. A central domain of 155 amino acid residues contains the Cys/His catalytic dyad and is the most conserved region containing at least 30 amino acid identities in all metacaspases. The Acanthamoeba castellanii metacaspase has the most proline-rich N-terminus so far reported in type-1 metacaspases with over 40 prolines in the first 150 residues. Ala-Pro-Pro is present 11 times. Phylogenies constructed using only the conserved proteolytic domains or the complete sequences show identical branching patterns, differing only in the rates of change.

  9. Quasi two-dimensional astigmatic solitons in soft chiral metastructures.

    PubMed

    Laudyn, Urszula A; Jung, Paweł S; Karpierz, Mirosław A; Assanto, Gaetano

    2016-03-15

    We investigate a non-homogeneous layered structure encompassing dual spatial dispersion: continuous diffraction in one transverse dimension and discrete diffraction in the orthogonal one. Such dual diffraction can be balanced out by one and the same nonlinear response, giving rise to light self-confinement into astigmatic spatial solitons: self-focusing can compensate for the spreading of a bell-shaped beam, leading to quasi-2D solitary wavepackets which result from 1D transverse self-localization combined with a discrete soliton. We demonstrate such intensity-dependent beam trapping in chiral soft matter, exhibiting one-dimensional discrete diffraction along the helical axis and one-dimensional continuous diffraction in the orthogonal plane. In nematic liquid crystals with suitable birefringence and chiral arrangement, the reorientational nonlinearity is shown to support bell-shaped solitary waves with simple astigmatism dependent on the medium birefringence as well as on the dual diffraction of the input wavepacket. The observations are in agreement with a nonlinear nonlocal model for the all-optical response.

  10. Quasi two-dimensional astigmatic solitons in soft chiral metastructures

    NASA Astrophysics Data System (ADS)

    Laudyn, Urszula A.; Jung, Paweł S.; Karpierz, Mirosław A.; Assanto, Gaetano

    2016-03-01

    We investigate a non-homogeneous layered structure encompassing dual spatial dispersion: continuous diffraction in one transverse dimension and discrete diffraction in the orthogonal one. Such dual diffraction can be balanced out by one and the same nonlinear response, giving rise to light self-confinement into astigmatic spatial solitons: self-focusing can compensate for the spreading of a bell-shaped beam, leading to quasi-2D solitary wavepackets which result from 1D transverse self-localization combined with a discrete soliton. We demonstrate such intensity-dependent beam trapping in chiral soft matter, exhibiting one-dimensional discrete diffraction along the helical axis and one-dimensional continuous diffraction in the orthogonal plane. In nematic liquid crystals with suitable birefringence and chiral arrangement, the reorientational nonlinearity is shown to support bell-shaped solitary waves with simple astigmatism dependent on the medium birefringence as well as on the dual diffraction of the input wavepacket. The observations are in agreement with a nonlinear nonlocal model for the all-optical response.

  11. Quasi two-dimensional astigmatic solitons in soft chiral metastructures

    PubMed Central

    Laudyn, Urszula A.; Jung, Paweł S.; Karpierz, Mirosław A.; Assanto, Gaetano

    2016-01-01

    We investigate a non-homogeneous layered structure encompassing dual spatial dispersion: continuous diffraction in one transverse dimension and discrete diffraction in the orthogonal one. Such dual diffraction can be balanced out by one and the same nonlinear response, giving rise to light self-confinement into astigmatic spatial solitons: self-focusing can compensate for the spreading of a bell-shaped beam, leading to quasi-2D solitary wavepackets which result from 1D transverse self-localization combined with a discrete soliton. We demonstrate such intensity-dependent beam trapping in chiral soft matter, exhibiting one-dimensional discrete diffraction along the helical axis and one-dimensional continuous diffraction in the orthogonal plane. In nematic liquid crystals with suitable birefringence and chiral arrangement, the reorientational nonlinearity is shown to support bell-shaped solitary waves with simple astigmatism dependent on the medium birefringence as well as on the dual diffraction of the input wavepacket. The observations are in agreement with a nonlinear nonlocal model for the all-optical response. PMID:26975651

  12. Influence of Acanthamoeba genotype on clinical course and outcomes for patients with Acanthamoeba keratitis in Spain.

    PubMed

    Arnalich-Montiel, Francisco; Lumbreras-Fernández, Blanca; Martín-Navarro, Carmen M; Valladares, Basilio; Lopez-Velez, Rogelio; Morcillo-Laiz, Rafael; Lorenzo-Morales, Jacob

    2014-04-01

    Genotype T4 is by far the most frequent genotype of Acanthamoeba keratitis (AK) and therefore has been considered the most virulent. This study included 14 cases of AK of genotype T4 and three cases of non-T4 genotype. We found that cases of non-T4 genotype had a worse response to medical therapy, greater need for surgical intervention, greater risk of extracorneal involvement, and remarkably poorer final visual outcome than those of T4 genotype, suggesting an association between Acanthamoeba virulence and genotype that requires additional case investigation.

  13. Influence of Acanthamoeba Genotype on Clinical Course and Outcomes for Patients with Acanthamoeba Keratitis in Spain

    PubMed Central

    Lumbreras-Fernández, Blanca; Martín-Navarro, Carmen M.; Valladares, Basilio; Lopez-Velez, Rogelio; Morcillo-Laiz, Rafael; Lorenzo-Morales, Jacob

    2014-01-01

    Genotype T4 is by far the most frequent genotype of Acanthamoeba keratitis (AK) and therefore has been considered the most virulent. This study included 14 cases of AK of genotype T4 and three cases of non-T4 genotype. We found that cases of non-T4 genotype had a worse response to medical therapy, greater need for surgical intervention, greater risk of extracorneal involvement, and remarkably poorer final visual outcome than those of T4 genotype, suggesting an association between Acanthamoeba virulence and genotype that requires additional case investigation. PMID:24430449

  14. Phagocytosis Affects Biguanide Sensitivity of Acanthamoeba spp.

    PubMed Central

    Noble, Judith A.; Ahearn, Donald G.; Avery, Simon V.; Crow Jr., Sidney A.

    2002-01-01

    The incidence of Acanthamoeba keratitis, a disease associated with contact lens wear, has been in apparent decline with the advent of multipurpose contact lens solutions. The concentrations of the biguanides chlorhexidine digluconate (CHX) and particularly polyhexamethylene biguanide (PHMB) included in multipurpose solutions (MPSs) are sublethal for amoebae. We evaluated by flow cytometry the effects of these two biguanides on phagocytosis of particles and the survival of trophozoites of Acanthamoeba castellanii and A. polyphaga. Trophozoites of A. castellanii and A. polyphaga (106/ml) were exposed to solutions of 5 and 50 μg of PHMB and CHX per ml in the presence and absence of particles (i.e., heat-killed yeasts and bacteria and latex beads). In addition, trophozoites were exposed to particles treated with these concentrations of the two biguanides. In the absence of particles, trophozoites of A. polyphaga appeared to be more resistant to the biguanides than those of A. castellanii. In the presence of particles, the rates of survival of both species were decreased. In most instances, particles treated with sublethal concentrations of both biguanides that were adsorbed onto the particles reduced the incidence of phagocytosis. Particles present in MPSs in contact lens cases may be involved in the decreased incidence of Acanthamoeba keratitis. PMID:12069957

  15. Phagocytosis affects biguanide sensitivity of Acanthamoeba spp.

    PubMed

    Noble, Judith A; Ahearn, Donald G; Avery, Simon V; Crow, Sidney A

    2002-07-01

    The incidence of Acanthamoeba keratitis, a disease associated with contact lens wear, has been in apparent decline with the advent of multipurpose contact lens solutions. The concentrations of the biguanides chlorhexidine digluconate (CHX) and particularly polyhexamethylene biguanide (PHMB) included in multipurpose solutions (MPSs) are sublethal for amoebae. We evaluated by flow cytometry the effects of these two biguanides on phagocytosis of particles and the survival of trophozoites of Acanthamoeba castellanii and A. polyphaga. Trophozoites of A. castellanii and A. polyphaga (10(6)/ml) were exposed to solutions of 5 and 50 microg of PHMB and CHX per ml in the presence and absence of particles (i.e., heat-killed yeasts and bacteria and latex beads). In addition, trophozoites were exposed to particles treated with these concentrations of the two biguanides. In the absence of particles, trophozoites of A. polyphaga appeared to be more resistant to the biguanides than those of A. castellanii. In the presence of particles, the rates of survival of both species were decreased. In most instances, particles treated with sublethal concentrations of both biguanides that were adsorbed onto the particles reduced the incidence of phagocytosis. Particles present in MPSs in contact lens cases may be involved in the decreased incidence of Acanthamoeba keratitis.

  16. Current Status of Acanthamoeba in Iran: A Narrative Review Article

    PubMed Central

    NIYYATI, Maryam; REZAEIAN, Mostafa

    2015-01-01

    Background: Free-living amoebae belonging to the genus Acanthamoeba have an environmental distribution. Amoebic keratitis due to these protozoan parasites continue to rise in Iran and worldwide. In Iran, there are various researches regarding both morphological and molecular identification of Acanthamoeba spp. in environmental and clinical samples. However, there is no thorough review about Acanthamoeba genotypes and their distribution in environmental sources such as water, dust and biofilm in Iran. Besides, according to increasing cases of Amoebic keratitis in the region awareness regarding the pathogenic potential of these sight-threatening amoebae is of utmost importance. Methods: We conducted a thorough review based on the database sources such as MEDLINE, PubMed and Google scholar. No restrictions were placed on study date, study design or language of publication. We searched all valuable and relevant information considering the occurrence of the Acanthamoeba in both environmental and clinical samples. Results: According to our thorough review Acanthamoeba belonging to T4 genotype is the most prevalent type strain in environmental and clinical samples in several regions in Iran and worldwide, however, there are reports regarding Acanthamoeba belonging to other genotypes such as T2, T3, T5, T6 and T11 and the mentioned point could leads us to more researches with the goal of presenting the real genotype dominance of Acanthamoeba and related disease in the country. Conclusion: Overall, the present review will focus on present status of genotypes of Acanthamoeba in Iran during recent years. PMID:26246812

  17. Endosymbionts of Acanthamoeba isolated from domestic tap water in Korea.

    PubMed

    Choi, Seon Hee; Cho, Min Kyoung; Ahn, Soon Cheol; Lee, Ji Eun; Lee, Jong Soo; Kim, Dong-Hee; Xuan, Ying-Hua; Hong, Yeon Chul; Kong, Hyun Hee; Chung, Dong Il; Yu, Hak Sun

    2009-12-01

    In a previous study, we reported our discovery of Acanthamoeba contamination in domestic tap water; in that study, we determined that some Acanthamoeba strains harbor endosymbiotic bacteria, via our molecular characterization by mitochondrial DNA restriction fragment length polymorphism (Mt DNA RFLP). Five (29.4%) among 17 Acanthamoeba isolates contained endosymbionts in their cytoplasm, as demonstrated via orcein staining. In order to estimate their pathogenicity, we conducted a genetic characterization of the endosymbionts in Acanthamoeba isolated from domestic tap water via 16S rDNA sequencing. The endosymbionts of Acanthamoeba sp. KA/WP3 and KA/WP4 evidenced the highest level of similarity, at 97% of the recently published 16S rDNA sequence of the bacterium, Candidatus Amoebophilus asiaticus. The endosymbionts of Acanthamoeba sp. KA/WP8 and KA/WP12 shared a 97% sequence similarity with each other, and were also highly similar to Candidatus Odyssella thessalonicensis, a member of the alpha-proteobacteria. The endosymbiont of Acanthamoeba sp. KA/WP9 exhibits a high degree of similarity (85-95%) with genus Methylophilus, which is not yet known to harbor any endosymbionts. This is the first report, to the best of our knowledge, to show that Methylophilus spp. can live in the cytoplasm of Acanthamoeba.

  18. Cytotoxic effect of acriflavine against clinical isolates of Acanthamoeba spp.

    PubMed

    Polat, Zubeyda Akin; Karakus, Gulderen

    2013-02-01

    Acanthamoeba keratitis (AK) is a potentially devastating and sight-threatening infection of the cornea caused by the ubiquitous free-living amoebae, Acanthamoeba species. Its eradication is difficult because the amoebas encyst, making it highly resistant to anti-amoebic drugs. Acriflavine neutral (ACF) has been used for treatment of microbial infections for humans and fishes. The aim of our study was to evaluate the time-dependent cytotoxicities of ACF against Acanthamoeba spp. Trophozoites and cysts of three different strains (strain PAT06 Acanthamoeba castellanii, strain 2HH Acanthamoeba hatchetti, and strain 11DS A. hatchetti) of Acanthamoeba spp. were tested. All strains had been isolated from patients suffering from a severe AK. The effects of the ACF with the concentrations ranging from 15 to 500 mg mL(-1) on the cytotoxicity of Acanthamoeba strains were examined. ACF showed a time- and dose-dependent amebicidal action on the trophozoites and cysts. Pat06 (A. castellanii) was the most resistant, while strain 11DS (A. hatchetti) was the most sensitive. As a result, ACF could be concluded as a new agent for the treatment of Acanthamoeba infections. On the other hand, it still needs to be further evaluated by in vivo test systems to confirm the efficiency of its biological effect.

  19. Comparative evaluation of the short-term bactericidal potential of a steroid-antibiotic combination versus steroid in the treatment of chronic bacterial blepharitis and conjunctivitis.

    PubMed

    Shulman, D G; Sargent, J B; Stewart, R H; Mester, U

    1996-01-01

    The effects of four days' treatment with topical Maxitrol (neomycin sulphate 3500 IU/mL, polymyxin-B sulphate 6000 IU/mL with dexamethasone 0.1%) were compared with those of Maxidex (dexamethasone 0.1% alone) in a double-masked study in 111 patients with bacterial blepharitis or conjunctivitis, 95 of whom were evaluable for efficacy. The majority of patients (N = 80) had chronic blepharitis. Maxitrol treatment resulted in a significantly greater reduction (90%) in bacterial counts and bacterial eradication (50%) compared with Maxidex (34% and 17% respectively). Maxitrol treatment also produced a significantly greater reduction in conjunctival discharge than did Maxidex, while the treatments were equally effective in alleviating other ocular signs and symptoms. It was concluded that use of a fixed dose combination steroid-antibiotic product was more effective for bacterial control and therapeutic efficacy in the treatment of chronic blepharitis and conjunctivitis patients than treatment with steroid alone. However, in the long-term treatment of chronic blepharitis the well-known toxic problems of neomycin sulphate have to be taken into account.

  20. Occurrence of Potentially Pathogenic Bacterial-Endosymbionts in Acanthamoeba Spp.

    PubMed Central

    NIYYATI, Maryam; MAFI, Mahyar; HAGHIGHI, Ali; HAKEMI VALA, Mojdeh

    2015-01-01

    Background: Acanthamoeba- bacteria interactions enable pathogenic bacteria to tolerate harsh conditions and lead to transmission to the susceptible host. The present study was aimed to address the presence of bacterial endosymbionts of Acanthamoeba isolated from recreational water sources of Tehran, Iran. To the best of our knowledge this is the first study regarding occurrence of bacteria in environmental Acanthamoeba spp. in Iran. Methods: A total of 75 samples of recreational water sources were collected. Samples were cultured on non- nutrient agar 1.5% plates. Positive Acanthamoeba spp. were axenically grown. DNA extraction and PCR reaction was performed using JDP1-2 primers. All positive samples of Acanthamoeba were examined for the presence of endosymbionts using staining and molecular methods. The PCR products were then sequenced in order to determine the genotypes of Acanthamoeba and bacteria genera. Results: Out of 75 samples, 16 (21.3%) plates were positive for Acanthamoeba according to the morphological criteria. Molecular analysis revealed that Acanthamoeba belonged to T4 and T5 genotypes. Five isolates (35.7%) were positive for bacterial endosymbionts using staining method and PCR test. Sequencing of PCR products confirmed the presence of Pseudomonas aeruginosa and Agrobacterium tumefasiens. Conclusion: The presence of Acanthamoeba bearing pathogenic endosymbionts in water sources leads us to public health issues including improved sanitation and decontamination measures in recreational water sources in order to prevent amoebae-related infection. To the best of our knowledge this is the first report regarding the isolation of A. tumefasiens from Acanthamoeba in Iran and worldwide. PMID:26246815

  1. Acanthamoeba induces cell-cycle arrest in host cells.

    PubMed

    Sissons, James; Alsam, Selwa; Jayasekera, Samantha; Kim, Kwang Sik; Stins, Monique; Khan, Naveed Ahmed

    2004-08-01

    Acanthamoeba can cause fatal granulomatous amoebic encephalitis (GAE) and eye keratitis. However, the pathogenesis and pathophysiology of these emerging diseases remain unclear. In this study, the effects of Acanthamoeba on the host cell cycle using human brain microvascular endothelial cells (HBMEC) and human corneal epithelial cells (HCEC) were determined. Two isolates of Acanthamoeba belonging to the T1 genotype (GAE isolate) and T4 genotype (keratitis isolate) were used, which showed severe cytotoxicity on HBMEC and HCEC, respectively. No tissue specificity was observed in their ability to exhibit binding to the host cells. To determine the effects of Acanthamoeba on the host cell cycle, a cell-cycle-specific gene array was used. This screened for 96 genes specific for host cell-cycle regulation. It was observed that Acanthamoeba inhibited expression of genes encoding cyclins F and G1 and cyclin-dependent kinase 6, which are proteins important for cell-cycle progression. Moreover, upregulation was observed of the expression of genes such as GADD45A and p130 Rb, associated with cell-cycle arrest, indicating cell-cycle inhibition. Next, the effect of Acanthamoeba on retinoblastoma protein (pRb) phosphorylation was determined. pRb is a potent inhibitor of G1-to-S cell-cycle progression; however, its function is inhibited upon phosphorylation, allowing progression into S phase. Western blotting revealed that Acanthamoeba abolished pRb phosphorylation leading to cell-cycle arrest at the G1-to-S transition. Taken together, these studies demonstrated for the first time that Acanthamoeba inhibits the host cell cycle at the transcriptional level, as well as by modulating pRb phosphorylation using host cell-signalling mechanisms. A complete understanding of Acanthamoeba-host cell interactions may help in developing novel strategies to treat Acanthamoeba infections.

  2. Variation of axial and oblique astigmatism with accommodation across the visual field

    PubMed Central

    Liu, Tao; Thibos, Larry N.

    2017-01-01

    In this study we investigated the impact of accommodation on axial and oblique astigmatism along 12 meridians of the central 30° of visual field and explored the compensation of corneal first-surface astigmatism by the remainder of the eye's optical system. Our experimental evidence revealed no systematic effect of accommodation on either axial or oblique astigmatism for two adult populations (myopic and emmetropic eyes). Although a few subjects exhibited systematic changes in axial astigmatism during accommodation, the dioptric value of these changes was much smaller than the amount of accommodation. For most subjects, axial and oblique astigmatism of the whole eye are both less than for the cornea alone, which indicates a compensatory role for internal optics at all accommodative states in both central and peripheral vision. A new method for determining the eye's optical axis based on visual field maps of oblique astigmatism revealed that, on average, the optical axis is 4.8° temporal and 0.39° superior to the foveal line-of-sight in object space, which agrees with previous results obtained by different methodologies and implies that foveal astigmatism includes a small amount of oblique astigmatism (0.06 D on average). Customized optical models of each eye revealed that oblique astigmatism of the corneal first surface is negligible along the pupillary axis for emmetropic and myopic eyes. Individual variation in the eye's optical axis is due in part to misalignment of the corneal and internal components that is consistent with tilting of the crystalline lens relative to the pupillary axis. PMID:28362902

  3. In vitro comparative assessment of different viability assays in Acanthamoeba castellanii and Acanthamoeba polyphaga trophozoites.

    PubMed

    Heredero-Bermejo, I; Copa-Patiño, J L; Soliveri, J; Gómez, R; de la Mata, F J; Pérez-Serrano, J

    2013-12-01

    The species of the genus Acanthamoeba are opportunistic protozoan parasites that cause different diseases in humans, such as amoebic keratitis and granulomatous encephalitis. The rise in the rate of Acanthamoeba keratitis, mainly due to the increase in contact lens wearers, turns the development of viability assays using a multi-well plate reader as a tool for screening new antiamoebic agents in vitro into an important goal. In our study, the viability assays PrestoBlue®, resazurin sodium salt, 3-(4,5-dimethylthiazol-2yl)-2,5-diphenyltetrazolium bromide (MTT) and CellTiter96® were tested for their suitability as time-saving alternatives to the classical manual or direct-counting method, assessing the effect of the antiamoebic agent chlorhexidine digluconate and temperature on Acanthamoeba castellanii (ATCC® 30234™) and Acanthamoeba polyphaga 2961. Although resazurin and MTT have already been previously used in amoeba viability assays to test the activities of antiamoebic agents in vitro, it is the first time that PrestoBlue® and CellTiter96® are used for this purpose. Results indicated that the viability assays were strain-dependent leading in some cases to an overestimation of the real situation of viable cells. This implies that each viability assay ought to be set up for each amoeba strain studied.

  4. Proximity- and Astigmatism-Tolerant Testsites For Electrical Linewidth Measurement

    NASA Astrophysics Data System (ADS)

    Lin, Burn J.

    1989-07-01

    Electrical linewidth measurement is well-known for high precision and throughput. However, the standard four-point probe testsite is only useful for measuring the width of an isolated conducting line. Line-and-space and isolated spaces can be simulated satisfactorily by adding dummy lines parallel to the active line but weak links or potential electrical shorting situations often prematurely cause these structures to fail before their true limits are reached. In this paper, fully wrapped proximity- and astigmatism-tolerant designs for line-and-space and isolated spaces are shown. They have been successfully demonstrated with printed images. An application in evaluating the exposure-defocus window of a one-layer i-line resist using the proximity-tolerant testsites is given.

  5. An update on Acanthamoeba keratitis: diagnosis, pathogenesis and treatment

    PubMed Central

    Lorenzo-Morales, Jacob; Khan, Naveed A.; Walochnik, Julia

    2015-01-01

    Free-living amoebae of the genus Acanthamoeba are causal agents of a severe sight-threatening infection of the cornea known as Acanthamoeba keratitis. Moreover, the number of reported cases worldwide is increasing year after year, mostly in contact lens wearers, although cases have also been reported in non-contact lens wearers. Interestingly, Acanthamoeba keratitis has remained significant, despite our advances in antimicrobial chemotherapy and supportive care. In part, this is due to an incomplete understanding of the pathogenesis and pathophysiology of the disease, diagnostic delays and problems associated with chemotherapeutic interventions. In view of the devastating nature of this disease, here we present our current understanding of Acanthamoeba keratitis and molecular mechanisms associated with the disease, as well as virulence traits of Acanthamoeba that may be potential targets for improved diagnosis, therapeutic interventions and/or for the development of preventative measures. Novel molecular approaches such as proteomics, RNAi and a consensus in the diagnostic approaches for a suspected case of Acanthamoeba keratitis are proposed and reviewed based on data which have been compiled after years of working on this amoebic organism using many different techniques and listening to many experts in this field at conferences, workshops and international meetings. Altogether, this review may serve as the milestone for developing an effective solution for the prevention, control and treatment of Acanthamoeba infections. PMID:25687209

  6. An update on Acanthamoeba keratitis: diagnosis, pathogenesis and treatment.

    PubMed

    Lorenzo-Morales, Jacob; Khan, Naveed A; Walochnik, Julia

    2015-01-01

    Free-living amoebae of the genus Acanthamoeba are causal agents of a severe sight-threatening infection of the cornea known as Acanthamoeba keratitis. Moreover, the number of reported cases worldwide is increasing year after year, mostly in contact lens wearers, although cases have also been reported in non-contact lens wearers. Interestingly, Acanthamoeba keratitis has remained significant, despite our advances in antimicrobial chemotherapy and supportive care. In part, this is due to an incomplete understanding of the pathogenesis and pathophysiology of the disease, diagnostic delays and problems associated with chemotherapeutic interventions. In view of the devastating nature of this disease, here we present our current understanding of Acanthamoeba keratitis and molecular mechanisms associated with the disease, as well as virulence traits of Acanthamoeba that may be potential targets for improved diagnosis, therapeutic interventions and/or for the development of preventative measures. Novel molecular approaches such as proteomics, RNAi and a consensus in the diagnostic approaches for a suspected case of Acanthamoeba keratitis are proposed and reviewed based on data which have been compiled after years of working on this amoebic organism using many different techniques and listening to many experts in this field at conferences, workshops and international meetings. Altogether, this review may serve as the milestone for developing an effective solution for the prevention, control and treatment of Acanthamoeba infections.

  7. Diversity and seasonal impact of Acanthamoeba species in a subtropical rivershed.

    PubMed

    Kao, Po-Min; Chou, Ming-Yuan; Tao, Chi-Wei; Huang, Wen-Chien; Hsu, Bing-Mu; Shen, Shu-Min; Fan, Cheng-Wei; Chiu, Yi-Chou

    2013-01-01

    This study evaluated the presence of Acanthamoeba species in the Puzih River watershed, which features typical subtropical monsoon climate and is located just above the Tropic of Cancer in Taiwan. The relationship between the seasonal and geographical distributions of Acanthamoeba species in this rivershed was also investigated. Acanthamoeba species were detected in water samples using the amoebal enrichment culture method and confirmed by PCR. A total of 136 water samples were included in this study, 16 (11.7%) of which contained Acanthamoeba species. Samples with the highest percentage of Acanthamoeba (32.4%) were obtained during the summer season, mainly from upstream areas. The identified species in the four seasons included Acanthamoeba palestinensis (T2), Acanthamoeba sp. IS2/T4 (T4), Acanthamoeba lenticulata (T5), Acanthamoeba hatchetti (T11), Acanthamoeba healyi (T12), and Acanthamoeba jacobsi (T15). The most frequently identified Acanthamoeba genotype was T4 (68.7%). Acanthamoeba genotype T4 is responsible for Acanthamoeba keratitis and should be considered for associated human health risk potential in the rivershed.

  8. Predator vs aliens: bacteria interactions with Acanthamoeba.

    PubMed

    Khan, Naveed Ahmed; Siddiqui, Ruqaiyyah

    2014-06-01

    By interactions with other microbes, free-living amoebae play a significant role in microbiology, environmental biology, physiology, cellular interactions, ecology and evolution. Here, we discuss astonishing interactions of bacteria and amoebae, in the light of evolution and functional aspects impacting human health. In favourable environmental conditions, the interaction of Acanthamoeba with non-virulent bacteria results in lysis of the bacteria. However, the interaction with weak-virulent bacteria results in a symbiotic relationship or amoebal lysis may occur. The microbial survival of amoebae in harsh environments, ability to interact with bacteria, and their ability to aid transmission to susceptible hosts is of great concern to human, animal and ecosystem health.

  9. On astigmatism of multi-beam optical stress sensor mounted at large incident angle

    NASA Astrophysics Data System (ADS)

    Jiang, Jinbo; Hwang, Heedon; Lee, Hak Sun; Kim, Byongju; Bong, Kee; Yoon, Euijoon

    2004-01-01

    When multi-beam optical stress sensor (MOSS) system is mounted at a large incident angle ( α), despite an improvement of the resolution in the measurements, it also induces optical astigmatism in the spot images on a charge-coupled device. During epitaxial growth, as the film stress increases, the astigmatism may result in the beam deflection changing at different rates in the directions parallel and perpendicular to the incident plane, if the α is large. In this paper, the system error due to the astigmatism is analyzed by the ray-tracing method and its predictions are compared with the experimental results. It is demonstrated here, how the spot spacing changes along the above mentioned orthogonal directions can be considered separately to minimize the error due to astigmatism in the MOSS measurements at any large α.

  10. On astigmatism of multi-beam optical stress sensor mounted at large incident angle

    NASA Astrophysics Data System (ADS)

    Jiang, Jinbo; Kee, Bong

    When multi-beam optical stress sensor (MOSS) system is mounted at a large incident angle ([alpha]), despite an improvement of the resolution in the measurements, it also induces optical astigmatism in the spot images on a charge coupled device (CCD). During epitaxial growth, as the film stress increases, the astigmatism may result in the spot spacing deflection ([delta]d) changing at different rates in the directions parallel and perpendicular to the incident plane, if the [alpha] is large. In this paper, the system error due to the astigmatism is analyzed by the ray tracing method and its predictions are compared with the the experimental results. It is demonstrated here, how the spot spacing deflections along the above mentioned orthogonal directions can be considered separately to minimize the error due to astigmatism in the MOSS measurements at any large [alpha].

  11. Three-dimensional location of micrometer-sized particles in macroscopic domains using astigmatic aberrations.

    PubMed

    Fuchs, Thomas; Hain, Rainer; Kähler, Christian J

    2014-03-01

    This Letter presents a theoretical and experimental image formation study in the presence of astigmatic aberrations. A three-dimensional, macroscopic location scheme of micrometer-sized particles for the single camera astigmatism particle tracking velocimetry (APTV) technique is introduced. Average particle z position determination errors of the technique are as low as 0.33%, with a measurement depth of 40 mm. These accuracies show APTV's ability of measuring volumetric velocity fields in macroscopic domains with limited optical access.

  12. Optical image reconstruction using an astigmatic lens for synthetic-aperture imaging ladar

    NASA Astrophysics Data System (ADS)

    Sun, Zhiwei; Hou, Peipei; Zhi, Yanan; Sun, Jianfeng; Zhou, Yu; Xu, Qian; Liu, Liren

    2014-11-01

    An optical processor for synthetic-aperture imaging ladar (SAIL) utilizing one astigmatic lens is proposed. The processor comprises two structures of transmitting and reflecting. The imaging process is mathematically analyzed using the unified data-collection equation of side-looking and down-looking SAILs. Results show that the astigmatic lens can be replaced with a cylindrical lens on certain conditions. To verify this concept, laboratory experiment is conducted, the imaging result of data collected from one SAIL demonstrator is given.

  13. Comparison of Astigmatism Induced by Combined Inferior Oblique Anterior Transposition Procedure and Lateral Rectus Recession Alone

    PubMed Central

    Eum, Sun Jung

    2016-01-01

    Purpose The purpose of this study is to compare the magnitude and axis of astigmatism induced by a combined inferior oblique (IO) anterior transposition procedure with lateral rectus (LR) recession versus LR recession alone. Methods Forty-six patients were retrospectively analyzed. The subjects were divided into two groups: those having concurrent inferior oblique muscle overaction (IOOA) and intermittent exotropia (group 1, 20 patients) and those having only intermittent exotropia as a control (group 2, 26 patients). Group 1 underwent combined anterior transposition of IO with LR recession and group 2 underwent LR recession alone. Induced astigmatism was defined as the difference between preoperative and postoperative astigmatism using double-angle vector analysis. Cylinder power, axis of induced astigmatism, and spherical equivalent were analyzed at 1 week, 1 month, and 3 months after surgery. Results Larger changes in the axis of induced astigmatism were observed in group 1, with 4.5° incyclotorsion, than in group 2 at 1 week after surgery (axis, 84.5° vs. 91°; p < 0.001). However, there was no statistically significant inter-group difference thereafter. Relaxation and rapid regression in the incyclotorsion of induced astigmatism were observed over-time. Spherical equivalent significantly decreased postoperatively at 1 month in both groups, indicating a myopic shift (p = 0.011 for group 1 and p = 0.019 for group 2) but did not show significant differences at 3 months after surgery (p = 0.107 for group 1 and p = 0.760 for group 2). Conclusions Combined IO anterior transposition procedures caused an increased change in the axis of induced astigmatism, including temporary incyclotorsion, during the first week after surgery. However, this significant difference was not maintained thereafter. Thus, combined IO surgery with LR recession does not seem to produce a sustained astigmatic change, which can be a potential risk factor of postoperative amblyopia or

  14. The efficacy of a single continuous nylon suture for control of post keratoplasty astigmatism.

    PubMed Central

    Van Meter, W

    1996-01-01

    PURPOSE: Post operative adjustment of a single continuous suture is an effective means of reducing post keratoplasty astigmatism. This study evaluates post keratoplasty keratometry following suture adjustment with an adjusted suture in place and after the suture is removed. METHODS: Average keratometric astigmatism was measured over 24 months time in 26 patients with an adjusted continuous suture and 24 patients with a continuous suture that was not adjusted. Average keratometry in 43 patients with an adjusted continuous suture was compared with 37 patients with combined continuous and interrupted sutures. Finally, suture out astigmatism in 19 adjusted patients was compared to six patients with no adjustment. RESULTS: There was an increase in average corneal astigmatism over two years of 2.2 diopters in the adjusted group and 1.7 diopters in the non-adjusted group with sutures in place. One year following surgery, average keratometry flattened from 47.5 to 42.9 diopters in the adjusted continuous group and from 47.0 to 46.0 diopters in the group with combined continuous and interrupted sutures. Following suture removal, average astigmatism in patients who had suture adjustment was 4.4 diopters +/- 2.5 diopters (range 1-10 diopters), and 6.01 diopters (range 4-7) in the non-adjusted group. CONCLUSIONS: Average post keratoplasty astigmatism increases after a continuous suture is adjusted but the increase is comparable to patients with acceptable astigmatism who do not require adjustment. More progressive corneal flattening over 12 months time is seen with a continuous suture than which combined sutures. Average suture out astigmatism was 4.0 diopters following suture adjustment, compared to an average of 8.4 diopters prior to adjustment. Images FIGURE 1 FIGURE 2 FIGURE 3 FIGURE 4 FIGURE 7A FIGURE 7B PMID:8981721

  15. A simple method for astigmatic compensation of folded resonator without Brewster window.

    PubMed

    Qiao, Wen; Xiaojun, Zhang; Yonggang, Wang; Liqun, Sun; Hanben, Niu

    2014-02-10

    A folded resonator requires an oblique angle of incidence on the folded curved mirror, which introduces astigmatic distortions that limit the performance of the lasers. We present a simple method to compensate the astigmatism of folded resonator without Brewster windows for the first time to the best of our knowledge. Based on the theory of the propagation and transformation of Gaussian beams, the method is both effective and reliable. Theoretical results show that the folded resonator can be compensated astigmatism completely when the following two conditions are fulfilled. Firstly, when the Gaussian beam with a determined size beam waist is obliquely incident on an off-axis concave mirror, two new Gaussian beam respectively in the tangential and sagittal planes are formed. Another off-axis concave mirror is located at another intersection point of the two new Gaussian beams. Secondly, adjusting the incident angle of the second concave mirror or its focal length can make the above two Gaussian beam coincide in the image plane of the second concave mirror, which compensates the astigmatic aberration completely. A side-pumped continues-wave (CW) passively mode locked Nd:YAG laser was taken as an example of the astigmatically compensated folded resonators. The experimental results show good agreement with the theoretical predictions. This method can be used effectively to design astigmatically compensated cavities resonator of high-performance lasers.

  16. Prevalence of Corneal Astigmatism in an NHS Cataract Surgery Practice in Northern Ireland

    PubMed Central

    Hassett, Patrick

    2017-01-01

    PURPOSE Post-operative corneal astigmatism following cataract surgery can leave the patient with visual impairment. Correcting it at the time of surgery with a toric intraocular lens (TIOL) can give patients a better final visual outcome. The purpose was to determine the prevalence of corneal astigmatism in a cataract population and assess the demand for TIOL. METHODS Keratometric data was collected and analyzed for all patients who attended for routine cataract surgery under the care of a single surgeon based in Altnagelvin Area Hospital, Northern Ireland (NI). All patients were included between January 2008 and December 2014. Data was collected retrospectively for this observational study. RESULTS There were 2080 consecutive eyes of 1788 patients. The mean corneal astigmatism was 1.09 ± 0.83. Corneal astigmatism was 1.50D or less in 1621 eyes (78%). It was more than 2.00 D in 242 eyes (11.6%), more than 2.50 D in 127 eyes (6.1%), more than 3.00D in 68 eyes (3.27%) and more than 3.50 D in 45 eyes (2.16%). CONCLUSION For routine cataract surgery, 41.3% of eyes had more than 1.00 D of corneal astigmatism and 11.6% had more and 2.00D. Females had more astigmatism than males. This shows the potential demand for the TIOL in this population.

  17. Comparison of Surgically Induced Astigmatisms after Clear Corneal Incisions of Different Sizes

    PubMed Central

    Mohamed, Tarek; Fine, I. Howard

    2007-01-01

    Purpose This study was performed to assess efficiency and stability of astigmatic change by incision size after cataract surgery. Methods This work was designed as a retrospective, comparative, nonrandomized interventional study. A total of 121 cases of cataract surgery were reviewed in 98 patients performed by one surgeon at the Oregon Eye Institute in Eugene, OR, USA with 3-year follow-ups. All procedures were performed with the temporal approach of self-sealing incisions. The serial change in surgically induced astigmatisms were examined in all cases of three groups: Group A, cartridge injection of a foldable IOL through a 2.5 mm self-sealing incision; Group B, cartridge injection of a foldable IOL through a 3.0 mm self-sealing incision; Group C, cartridge injection of a foldable IOL through a 3.5 mm self-sealing incision. Keratometric data were obtained reoperatively, and 3 weeks, 3 months, 6 months, 9 months, 12 months, 24 months and 36 months postoperatively. Polar value analysis was performed to calculate the surgically induced astigmatism. Results The astigmatic change decreased over time in Group B (P<0.05). The other groups tended to remain in induced astigmatism. All groups showed anticlockwise torque at 3 weeks following surgery. Group B showed a decrease in deviation, but the other groups showed increases in their torque value at postoperative 12 months (P<0.05). Conclusions The 3.0 mm incision size correlated with the least surgically induced astigmatism. PMID:17460424

  18. [Acanthamoeba spp. as opportunistic pathogens parasites].

    PubMed

    Castrillón, Juan C; Orozco, Lina P

    2013-04-01

    Among free-living amoeba in nature, species of the genus Acanthamoeba have been associated with human disease. These amoeba are among the most abundant protozoa in nature due to its cosmopolitan distribution and are able to survive in a wide variety of habitats because its low demand for food and in harsh environments by forming structures known as cysts. However, ecological changes and incursion of its different habitats have made this organism can invade a host and live as parasites within him. That's why this type of protozoa are known as amphizoic organism, because human can be constituted as its host, causing infections in the central nervous system, disseminated infections in skin and lungs, and keratitis. Thus, since an increase in the number of cases of Acanthamoeba infections has occurred worldwide, these protozoa have become increasingly important as agents of human disease. This review summarizes what is known of this kind of free-living amoeba, focusing on the biology, ecology, pathogenesis, diagnosis, treatment and human defense mechanism against infection by the amoeba.

  19. Gene discovery in the Acanthamoeba castellanii genome

    SciTech Connect

    Anderson, Iain J.; Watkins, Russell F.; Samuelson, John; Spencer,David F.; Majoros, William H.; Gray, Michael W.; Loftus, Brendan J.

    2005-08-01

    Acanthamoeba castellanii is a free-living amoeba found in soil, freshwater, and marine environments and an important predator of bacteria. Acanthamoeba castellanii is also an opportunistic pathogen of clinical interest, responsible for several distinct diseases in humans. In order to provide a genomic platform for the study of this ubiquitous and important protist, we generated a sequence survey of approximately 0.5 x coverage of the genome. The data predict that A. castellanii exhibits a greater biosynthetic capacity than the free-living Dictyostelium discoideum and the parasite Entamoeba histolytica, providing an explanation for the ability of A. castellanii to inhabit adversity of environments. Alginate lyase may provide access to bacteria within biofilms by breaking down the biofilm matrix, and polyhydroxybutyrate depolymerase may facilitate utilization of the bacterial storage compound polyhydroxybutyrate as a food source. Enzymes for the synthesis and breakdown of cellulose were identified, and they likely participate in encystation and excystation as in D. discoideum. Trehalose-6-phosphate synthase is present, suggesting that trehalose plays a role in stress adaptation. Detection and response to a number of stress conditions is likely accomplished with a large set of signal transduction histidine kinases and a set of putative receptorserine/threonine kinases similar to those found in E. histolytica. Serine, cysteine and metalloproteases were identified, some of which are likely involved in pathogenicity.

  20. Raman Microspectroscopy Analysis in the Treatment of Acanthamoeba Keratitis

    PubMed Central

    Rusciano, Giulia; Capriglione, Paola; Pesce, Giuseppe; Del Prete, Salvatore; Cennamo, Gilda; Di Cave, David; Cerulli, Luciano; Sasso, Antonio

    2013-01-01

    Acanthamoeba keratitis is a rare but serious corneal disease, often observed in contact lens wearers. Clinical treatment of infected patients frequently involves the use of polyhexamethylene biguanide (PHMB), a polymer used as a disinfectant and antiseptic, which is toxic also for the epithelial cells of the cornea. Prompt and effective diagnostic tools are hence highly desiderable for both starting early therapy and timely suspension of the treatment. In this work we use Raman microspectroscopy to analyse in vitro a single Acanthamoeba cell in cystic phase. In particular, we investigate the effect of PHMB at the single-cell level, providing useful information on both the underlying biochemical mechanism and the time frame for Acanthamoeba eradication in ocular infections. Furthermore, we demonstrate that Raman spectroscopy, in conjunction with standard multivariate analysis methods, allows discriminating between live and dead Acanthamoebas, which is fundamental to optimizing patients’ treatment. PMID:23977228

  1. Advances in the Diagnosis and Treatment of Acanthamoeba Keratitis

    PubMed Central

    Clarke, Benjamin; Sinha, Arti; Parmar, Dipak N.

    2012-01-01

    This paper aims to review the recent literature describing Acanthamoeba keratitis and outline current thoughts on pathogenesis, diagnosis, and treatment as well as currently emerging diagnostic and treatment modalities. PMID:23304449

  2. Several staining techniques to enhance the visibility of Acanthamoeba cysts.

    PubMed

    El-Sayed, Nagwa Mostafa; Hikal, Wafaa Mohamed

    2015-03-01

    Acanthamoeba is one of the most common free-living amoebae. It is widespread in the environment and can infect humans causing keratitis. Delayed diagnosis or misdiagnosis leads to extensive corneal inflammation and profound visual loss. Therefore, accurate and rapid diagnosis of Acanthamoeba keratitis is essential for successful treatment and good prognosis. This study was designed to use different staining techniques to facilitate the identification of Acanthamoeba cysts. Acanthamoeba cysts were isolated by cultivation of either corneal scraping specimens or tap water samples onto non-nutrient agar plates seeded with Escherichia coli. Subcultures were done from positive cultures until unique cysts were isolated. Acanthamoeba cysts were stained temporarily using iodine, eosin, methylene blue, and calcofluor white (CFW) stains and as permanent slides after processing for mounting using modified trichrome, Gimenez and Giemsa staining. These stains were compared on the basis of staining quality including clarity of morphological details, differentiation between cytoplasm and nuclei, color and contrast, and also other characteristics of the staining techniques, including ease of handling, time taken for the procedure, and cost effectiveness. The cysts of Acanthamoeba were recognized in the form of double-walled cysts: the outer wall (ectocyst) that was being differentiated from the variably stained surrounding background and the inner wall (endocyst) that was sometimes stellated, polygonal, round, or oval and visualized as separate from the spherical, sometimes irregular, outline of the ectocyst. Regarding the temporary stains, it was found that they were efficient for visualizing the morphological details of Acanthamoeba cysts. In CFW staining, Acanthamoeba cysts appeared as bluish-white or turquoise oval halos although the internal detail was not evident. On the other hand, the results of permanent-stained slides showed the most consistent stain for identification of

  3. Crescent bodies of Parachlamydia acanthamoeba and its life cycle within Acanthamoeba polyphaga: an electron micrograph study.

    PubMed

    Greub, Gilbert; Raoult, Didier

    2002-06-01

    Parachlamydiaceae are endosymbionts of free-living amoeba first identified in 1997. Two developmental stages, elementary and reticulate bodies, were observed; however, their localization and proportions according to culture condition and duration remain unknown. The life cycle of Parachlamydia acanthamoeba within Acanthamoeba polyphaga was studied by transmission electron microscopy of 8-, 36-, and 144-h coculture. Morphometry and quantification were performed using SAMBA software. The elementary body, the predominant stage within the amoebae, was located mainly within their vacuoles. The multiplication of Parachlamydia bacteria by binary fission of reticulate bodies was independently associated with culture in PYG broth (odds ratio [OR] = 4.4; 95% confidence interval [CI], 1.55 to 12.46) and with the presence of reticulate bodies within the amoebae (OR = 2.10; 95% CI, 1.53 to 2.89). A third developmental stage was observed, the crescent body. Its presence outside and inside the amoebae was associated mainly with prolonged incubation time (OR = 3.98; 95% CI, 1.49 to 10.68, and OR = 5.98; 95% CI, 1.75 to 20.4, respectively). Elementary and crescent bodies were released into the extracellular medium within vesicles or after amoebal lysis. For both, phagocytosis was their mode of entry. This electron micrograph study revealed another infective developmental stage, the crescent body, and provided quantitative analysis of the life cycle of P. acanthamoeba within A. polyphaga.

  4. Crescent Bodies of Parachlamydia acanthamoeba and Its Life Cycle within Acanthamoeba polyphaga: an Electron Micrograph Study

    PubMed Central

    Greub, Gilbert; Raoult, Didier

    2002-01-01

    Parachlamydiaceae are endosymbionts of free-living amoeba first identified in 1997. Two developmental stages, elementary and reticulate bodies, were observed; however, their localization and proportions according to culture condition and duration remain unknown. The life cycle of Parachlamydia acanthamoeba within Acanthamoeba polyphaga was studied by transmission electron microscopy of 8-, 36-, and 144-h coculture. Morphometry and quantification were performed using SAMBA software. The elementary body, the predominant stage within the amoebae, was located mainly within their vacuoles. The multiplication of Parachlamydia bacteria by binary fission of reticulate bodies was independently associated with culture in PYG broth (odds ratio [OR] = 4.4; 95% confidence interval [CI], 1.55 to 12.46) and with the presence of reticulate bodies within the amoebae (OR = 2.10; 95% CI, 1.53 to 2.89). A third developmental stage was observed, the crescent body. Its presence outside and inside the amoebae was associated mainly with prolonged incubation time (OR = 3.98; 95% CI, 1.49 to 10.68, and OR = 5.98; 95% CI, 1.75 to 20.4, respectively). Elementary and crescent bodies were released into the extracellular medium within vesicles or after amoebal lysis. For both, phagocytosis was their mode of entry. This electron micrograph study revealed another infective developmental stage, the crescent body, and provided quantitative analysis of the life cycle of P. acanthamoeba within A. polyphaga. PMID:12039769

  5. Statins and Voriconazole Induce Programmed Cell Death in Acanthamoeba castellanii

    PubMed Central

    López-Arencibia, Atteneri; Sifaoui, Ines; Reyes-Batlle, María; Valladares, Basilio; Martínez-Carretero, Enrique; Piñero, José E.; Maciver, Sutherland K.; Lorenzo-Morales, Jacob

    2015-01-01

    Members of the genus Acanthamoeba are facultative pathogens of humans, causing a sight-threatening keratitis and a life-threatening encephalitis. In order to treat those infections properly, it is necessary to target the treatment not only to the trophozoite but also to the cyst. Furthermore, it may be advantageous to avoid parasite killing by necrosis, which may induce local inflammation. We must also avoid toxicity of host tissue. Many drugs which target eukaryotes are known to induce programmed cell death (PCD), but this process is poorly characterized in Acanthamoeba. Here, we study the processes of programmed cell death in Acanthamoeba, induced by several drugs, such as statins and voriconazole. We tested atorvastatin, fluvastatin, simvastatin, and voriconazole at the 50% inhibitory concentrations (IC50s) and IC90s that we have previously established. In order to evaluate this phenomenon, we investigated the DNA fragmentation, one of the main characteristics of PCD, with quantitative and qualitative techniques. Also, the changes related to phosphatidylserine exposure on the external cell membrane and cell permeability were studied. Finally, because caspases are key to PCD pathways, caspase activity was evaluated in Acanthamoeba. All the drugs assayed in this study induced PCD in Acanthamoeba. To the best of our knowledge, this is the first study where PCD induced by drugs is described quantitatively and qualitatively in Acanthamoeba. PMID:25733513

  6. Genotyping of clinical isolates of Acanthamoeba genus in Venezuela.

    PubMed

    Wagner, Carolina; Reyes-Batlle, María; Ysea, María Alejandra Vethencourt; Pérez, Mónica V Galindo; de Rondón, Carmen Guzmán; Paduani, Anaibeth J Nessi; Pérez, Angelyseb Dorta; López-Arencibia, Atteneri; Sifaoui, Ines; de Galindo, María Virginia Pérez; de Suárez, Eva Pérez; Martínez-Carretero, Enrique; Valladares, Basilio; Piñero, José E; Lorenzo-Morales, Jacob

    2016-12-01

    Free-living amoebae of Acanthamoeba genus are opportunistic pathogens distributed worldwide. Strains included in this genus are causative agents of a fatal encephalitis and a sight-threating keratitis in humans and other animals. In this study, 550 clinical samples which were collected between 1984 and 2014 from different patients with suspected infections due to Acanthamoeba were initially screened for the presence of this amoebic genus at the Laboratorio de Amibiasis-Escuela de Bioanálisis at the Universidad Central de Venezuela. Samples were cultured in 2% Non-Nutrient agar plates seeded with a layer of heat killed Escherichia coli. From the 550 clinical samples included in this study, 18 of them were positive for Acanthamoeba genus after culture identification. Moreover, positive samples were confirmed after amplification of the Diagnostic Fragment 3 (DF3) of the Acanthamoeba18S rDNA genus and sequencing was carried out in order to genotype the isolated strains of Acanthamoeba. Furthermore, the pathogenic potential of the strains was checked by performing thermotolerance and osmotolerance assays. Sequencing of the DF3 region resulted in the identification of genotype T4 in all the isolated strains. Moreover, most isolates were thermotolerant or both thermotolerant and osmotolerant and thus were classified as potentially pathogenic strains. To the best of our knowledge, this is the first report on the molecular characterization at the genotype level of Acanthamoeba strains in Venezuela.

  7. Prevalence of Corneal Astigmatism in Tohono O'odham Native American Children 6 Months to 8 Years of Age

    PubMed Central

    Dobson, Velma; Miller, Joseph M.; Schwiegerling, Jim; Clifford-Donaldson, Candice E.; Green, Tina K.; Messer, Dawn H.

    2011-01-01

    Purpose. To describe the prevalence of corneal astigmatism in infants and young children who are members of a Native American tribe with a high prevalence of refractive astigmatism. Methods. The prevalence of corneal astigmatism was assessed by obtaining infant keratometer (IK4) measurements from 1235 Tohono O'odham children, aged 6 months to 8 years. Results. The prevalence of corneal astigmatism >2.00 D was lower in the 1- to <2-year-old age group when compared with all other age groups, except the 6- to <7-year-old group. The magnitude of mean corneal astigmatism was significantly lower in the 1- to <2-year age group than in the 5- to <6-, 6- to <7-, and 7- to <8-year age groups. Corneal astigmatism was with-the-rule (WTR) in 91.4% of astigmatic children (≥1.00 D). Conclusions. The prevalence and mean amount of corneal astigmatism were higher than reported in non–Native American populations. Mean astigmatism increased from 1.43 D in 1-year-olds to nearly 2.00 D by school age. PMID:21460261

  8. Isolation of Acanthamoeba from the rhizosphere of maize and lucerne plants

    NASA Astrophysics Data System (ADS)

    Orosz, Erika; Farkas, Ágnes; Ködöböcz, László; Becsak, Péter; Danka, József; Kucsera, István; Füleky, György

    2013-04-01

    Acanthamoeba species are free-living amoebae that can be found in almost every range of environments. Within this genus, a number of species are recognized as human pathogens, potentially causing Acanthamoeba keratitis, granulomatous amoebic encephalitis, and chronic granulomatous lesions. Soil and water samples were taken from experimental station at Julianna Major of Plant Protection Institute of Centre for Agricultural Research, Hungarian Academy of Sciences. We detected living Acanthamoeba spp. based on culture- confirmed detection combined with the molecular taxonomic identification method. Living Acanthamoeba spp. were detected in thirteen (65%) samples. The presence of Acanthamoeba spp. in the samples depends significantly on the rhizosphere plants. The most frequently identified living Acanthamoeba genotype was T4 followed by T11, T2/T6 and T17. Genotypes T4 and T11 of Acanthamoeba, are responsible for Acanthamoeba keratitis as well as granulomatous amoebic encephalitis, and should therefore be considered as a potential health risk associated with human activities in the environment.

  9. A novel color-LED corneal topographer to assess astigmatism in pseudophakic eyes

    PubMed Central

    Ferreira, Tiago B; Ribeiro, Filomena J

    2016-01-01

    Purpose To assess the accuracy of corneal astigmatism evaluation measured by four techniques, Orbscan IIz®, Lenstar LS900®, Cassini®, and Total Cassini (anterior + posterior surface), in pseudophakic eyes. Patients and methods A total of 30 patients (46 eyes) who had undergone cataract surgery with the implantation of a monofocal intraocular lens (AcrySof IQ) were assessed after surgery. For each eye, subjective assessment of astigmatism and its axis was performed. Minimum, maximum, and mean keratometry and astigmatism and its axis were evaluated using the four measurement techniques. All measurements were compared with the subjective measurements. Agreement between each measurement technique and subjective assessment was evaluated using Bland–Altman plots. Linear regressions were performed and compared. Results Linear regression analysis of astigmatism axis showed very high R2 for all models, with Total Cassini showing the least difference to the unit slope (0.052) and the least difference to a null constant (3.790), although not statistically different from the other models. Regarding astigmatism value, the Cassini and Total Cassini models were similar and statistically better than the Lenstar model. Cassini and Total Cassini showed better J0 compared with Orbscan. Conclusion On linear regression models, Cassini and Total Cassini showed the best performance regarding astigmatism value. Cassini and Total Cassini also showed the least J0 deviation from the Cartesian origin compared with Orbscan, which had the lowest performance. Total corneal measurement with the color LED topographer seems to be a better technique for astigmatism assessment. PMID:27574391

  10. Encystment in Acanthamoeba castellanii: a review.

    PubMed

    Lloyd, David

    2014-11-01

    Differentiation of Acanthamoeba castellanii trophozoites involves massive turnover of cellular components and remodelling of organelle structure and function so as to produce a cryptobiotic cell, resistant to desiccation, heat, freezing, and chemical treatments. This review presents a summary of a decade of research on the most studied aspects of the biochemistry of this process, with emphasis on problems of biocide and drug resistances, putative new targets, molecular and cell biology of the process of encystment, and the characteristics of the encysted state. As well as the intrinsic pathogenicity of the organism towards the cornea, and the ability of related species to invade the human brain, its propensity for harbouring and transmitting pathogenic bacteria and viruses is considerable and leads to increasing concerns. The long-term survival and resistance of cysts to drugs and biocides adds another layer of complexity to the problem of their elimination.

  11. Acanthamoeba keratitis in the south of Sweden.

    PubMed

    Skarin, A; Florén, I; Kiss, K; Miörner, H; Stenevi, U

    1996-12-01

    Eight patients with Acanthamoeba keratitis were diagnosed and treated at our clinic between February 1991 and February 1993. Five of these were contact lens wearers, two had suffered recent corneal trauma and one had recently undergone penetrating keratoplasty. The diagnoses were based on both culture and histological examination of biopsy material in three cases, on culture alone in two cases and on histological examination alone in three cases. In all but one primary treatment was Propamidine isethionate and Neomycin/Polymyxin B topically and Ketoconazole orally. Because of poor healing three patients additionally received Paromomycin and Miconazole or Clotrimazol topically; two of these were further treated with Polyhexamethylene biguanide topically. The interval from initial symptoms to accurate diagnoses varied from one to eleven months. In one patient the eye could not be saved; in the remaining patients visual acuity after healing ranged from hand movements to 1.0.

  12. IDENTIFICATION OF Pseudomonas spp. AS AMOEBA-RESISTANT MICROORGANISMS IN ISOLATES OF Acanthamoeba

    PubMed Central

    Maschio, Vinicius José; Corção, Gertrudes; Rott, Marilise Brittes

    2015-01-01

    Acanthamoeba is a “Trojan horse” of the microbial world. The aim of this study was to identify the presence of Pseudomonas as an amoeba-resistant microorganism in 12 isolates of Acanthamoeba. All isolates showed the genus Pseudomonas spp. as amoeba-resistant microorganisms. Thus, one can see that the Acanthamoeba isolates studied are hosts of Pseudomonas. PMID:25651331

  13. identification of Pseudomonas spp. as amoeba-resistant microorganisms in isolates of Acanthamoeba.

    PubMed

    José Maschio, Vinicius; Corção, Gertrudes; Rott, Marilise Brittes

    2015-01-01

    Acanthamoeba is a "Trojan horse" of the microbial world. The aim of this study was to identify the presence of Pseudomonas as an amoeba-resistant microorganism in 12 isolates of Acanthamoeba. All isolates showed the genus Pseudomonas spp. as amoeba-resistant microorganisms. Thus, one can see that the Acanthamoeba isolates studied are hosts of Pseudomonas.

  14. Ecto-ATPases of clinical and non-clinical isolates of Acanthamoeba.

    PubMed

    Sissons, James; Alsam, Selwa; Jayasekera, Samantha; Khan, Naveed Ahmed

    2004-11-01

    Acanthamoeba are opportunistic protozoan parasites that can cause fatal granulomatous amoebic encephalitis and eye keratitis, however the pathogenic mechanisms of Acanthamoeba remain unclear. In this study, we described the ability of live Acanthamoeba to hydrolyse extracellular ATP. Both clinical and non-clinical isolates belonging to genotypes, T1, T2, T3, T4 and T7 exhibited ecto-ATPase activities in vitro. Using non-denaturing polyacrylamide gel electrophoresis, ecto-ATPases were further characterized. All Acanthamoeba isolates tested, exhibited a single ecto-ATPase band (approximate molecular weight of 272 kDa). However, clinical isolates exhibited additional bands suggesting that ecto-ATPases may play a role in the pathogenesis of Acanthamoeba. This was supported using suramin (ecto-ATPase inhibitor), which inhibited Acanthamoeba-induced host cell cytotoxicity. Previously, we and others have shown that Acanthamoeba binds to host cells using their mannose-binding protein and binding can be blocked using exogenous alpha-mannose. In this study, we observed that alpha-mannose significantly increased ecto-ATPase activities of pathogenic Acanthamoeba belonging to T1, T2, T3 and T4 genotypes but had no effect on non-pathogenic Acanthamoeba (belonging to T7 genotype). Overall, we have shown, for the first time, that Acanthamoeba exhibit ecto-ATPase activities, which may play a role in the pathogenesis of Acanthamoeba as well as their potential role in the differentiation of pathogenic Acanthamoeba.

  15. Acanthamoeba sohi, n. sp., a pathogenic Korean isolate YM-4 from a freshwater fish

    PubMed Central

    Im, Kyung-il

    2003-01-01

    A new species of Acanthamoeba was isolated from a freshwater fish in Korea and tentatively named Acanthamoeba sp. YM-4 (Korean isolate YM-4). The trophozoites were 11.0-23.0 µm in length and had hyaline filamentous projections. Cysts were similar to those of A. culbertsoni and A. royreba, which were previously designated as Acanthamoeba group III. Acanthamoeba YM-4 can survive at 40℃, and its generation time was 19.6 hr, which was longer than that of A. culbertsoni. In terms of the in vitro cytotoxicity of lysates, Acanthamoeba YM-4 was weaker than A. culbertsoni, but stronger than A. polyphaga. On the basis of the mortality of experimentally infected mice, Acanthamoeba YM-4 was found to be highly virulent. The isoenzymes profile of Acanthamoeba YM-4 was similar to that of A. royreba. An anti-Acanthamoeba YM-4 monoclonal antibody, McAY7, was found to react only with Acanthamoeba YM-4, and not with A. culbertsoni. Random amplified polymorphic DNA marker analysis and RFLP analysis of mitochondrial DNA and of 18S small subunit ribosomal RNA, placed Acanthamoeba YM-4 in a separate cluster on the basis of phylogenetic distances. Thus the Acanthamoeba Korean isolate YM-4 was identified as a new species, and assigned as Acanthamoeba sohi. PMID:14699258

  16. Applicability of supervised discriminant analysis models to analyze astigmatism clinical trial data

    PubMed Central

    Sedghipour, Mohammad Reza; Sadeghi-Bazargani, Homayoun

    2012-01-01

    Background In astigmatism clinical trials where more complex measurements are common, especially in nonrandomized small sized clinical trials, there is a demand for the development and application of newer statistical methods. Methods The source data belonged to a project on astigmatism treatment. Data were used regarding a total of 296 eyes undergoing different astigmatism treatment modalities: wavefront-guided photorefractive keratectomy, cross-cylinder photorefractive keratectomy, and monotoric (single) photorefractive keratectomy. Astigmatism analysis was primarily done using the Alpins method. Prior to fitting partial least squares regression discriminant analysis, a preliminary principal component analysis was done for data overview. Through fitting the partial least squares regression discriminant analysis statistical method, various model validity and predictability measures were assessed. Results The model found the patients treated by the wavefront method to be different from the two other treatments both in baseline and outcome measures. Also, the model found that patients treated with the cross-cylinder method versus the single method didn’t appear to be different from each other. This analysis provided an opportunity to compare the three methods while including a substantial number of baseline and outcome variables. Conclusion Partial least squares regression discriminant analysis had applicability for the statistical analysis of astigmatism clinical trials and it may be used as an adjunct or alternative analysis method in small sized clinical trials. PMID:23055670

  17. Convergence Insufficiency, Accommodative Insufficiency, Visual Symptoms, and Astigmatism in Tohono O'odham Students

    PubMed Central

    Twelker, J. Daniel; Miller, Joseph M.; Campus, Irene

    2016-01-01

    Purpose. To determine rate of convergence insufficiency (CI) and accommodative insufficiency (AI) and assess the relation between CI, AI, visual symptoms, and astigmatism in school-age children. Methods. 3rd–8th-grade students completed the Convergence Insufficiency Symptom Survey (CISS) and binocular vision testing with correction if prescribed. Students were categorized by astigmatism magnitude (no/low: <1.00 D, moderate: 1.00 D to <3.00 D, and high: ≥3.00 D), presence/absence of clinical signs of CI and AI, and presence of symptoms. Analyses determine rate of clinical CI and AI and symptomatic CI and AI and assessed the relation between CI, AI, visual symptoms, and astigmatism. Results. In the sample of 484 students (11.67 ± 1.81 years of age), rate of symptomatic CI was 6.2% and symptomatic AI 18.2%. AI was more common in students with CI than without CI. Students with AI only (p = 0.02) and with CI and AI (p = 0.001) had higher symptom scores than students with neither CI nor AI. Moderate and high astigmats were not at increased risk for CI or AI. Conclusions. With-the-rule astigmats are not at increased risk for CI or AI. High comorbidity rates of CI and AI and higher symptoms scores with AI suggest that research is needed to determine symptomatology specific to CI. PMID:27525112

  18. Distribution and Repeatability of Corneal Astigmatism Measurements (Magnitude and Axis) Evaluated With Color Light Emitting Diode Reflection Topography

    PubMed Central

    Asimellis, George

    2015-01-01

    Purpose: To evaluate and investigate the distribution and repeatability of anterior corneal surface astigmatism measurements (axis and magnitude) using a novel corneal topographer. Methods: Anterior corneal surface astigmatism was investigated in a total of 195 eyes using a novel multicolored spot reflection topographer (Cassini; i-Optics). Two patient groups were studied, a younger-age group A and an older-age group B. Three consecutive acquisitions were obtained from each eye. The repeatability of measurement was assessed using Bland–Altman plot analysis and is reported as the coefficient of repeatability. Results: Group A (average age 34.3 years) had on average with-the-rule astigmatism, whereas the older-age group B (average age 72.3 years) had on average against-the-rule astigmatism. Average astigmatism magnitude measurement repeatability in group A was 0.4 diopters (D) and in group B 0.4 D. Average astigmatism axis measurement repeatability in group A was 5.4 degrees and in group B 5.5 degrees. The axis measurement repeatability improved with increasing magnitude of astigmatism: in the subgroups with astigmatism between 3.0 and 6.0 D, the axis repeatability was 1.4 degrees (group A) and 1.2 degrees (group B), whereas in the subgroups with astigmatism larger than 6.0 D, the repeatability was 1.1 and 0.6 degrees, respectively. Conclusions: This novel corneal topography device seems to offer high precision in reporting corneal astigmatism. This study reaffirms the established trend of a corneal astigmatism shift from an average “with-the-rule” to “against-the-rule” with aging. PMID:26057324

  19. Acanthamoeba spp. as Agents of Disease in Humans

    PubMed Central

    Marciano-Cabral, Francine; Cabral, Guy

    2003-01-01

    Acanthamoeba spp. are free-living amebae that inhabit a variety of air, soil, and water environments. However, these amebae can also act as opportunistic as well as nonopportunistic pathogens. They are the causative agents of granulomatous amebic encephalitis and amebic keratitis and have been associated with cutaneous lesions and sinusitis. Immuno compromised individuals, including AIDS patients, are particularly susceptible to infections with Acanthamoeba. The immune defense mechanisms that operate against Acanthamoeba have not been well characterized, but it has been proposed that both innate and acquired immunity play a role. The ameba's life cycle includes an active feeding trophozoite stage and a dormant cyst stage. Trophozoites feed on bacteria, yeast, and algae. However, both trophozoites and cysts can retain viable bacteria and may serve as reservoirs for bacteria with human pathogenic potential. Diagnosis of infection includes direct microscopy of wet mounts of cerebrospinal fluid or stained smears of cerebrospinal fluid sediment, light or electron microscopy of tissues, in vitro cultivation of Acanthamoeba, and histological assessment of frozen or paraffin-embedded sections of brain or cutaneous lesion biopsy material. Immunocytochemistry, chemifluorescent dye staining, PCR, and analysis of DNA sequence variation also have been employed for laboratory diagnosis. Treatment of Acanthamoeba infections has met with mixed results. However, chlorhexidine gluconate, alone or in combination with propamidene isethionate, is effective in some patients. Furthermore, effective treatment is complicated since patients may present with underlying disease and Acanthamoeba infection may not be recognized. Since an increase in the number of cases of Acanthamoeba infections has occurred worldwide, these protozoa have become increasingly important as agents of human disease. PMID:12692099

  20. Acanthamoeba spp. as agents of disease in humans.

    PubMed

    Marciano-Cabral, Francine; Cabral, Guy

    2003-04-01

    Acanthamoeba spp. are free-living amebae that inhabit a variety of air, soil, and water environments. However, these amebae can also act as opportunistic as well as nonopportunistic pathogens. They are the causative agents of granulomatous amebic encephalitis and amebic keratitis and have been associated with cutaneous lesions and sinusitis. Immuno compromised individuals, including AIDS patients, are particularly susceptible to infections with Acanthamoeba. The immune defense mechanisms that operate against Acanthamoeba have not been well characterized, but it has been proposed that both innate and acquired immunity play a role. The ameba's life cycle includes an active feeding trophozoite stage and a dormant cyst stage. Trophozoites feed on bacteria, yeast, and algae. However, both trophozoites and cysts can retain viable bacteria and may serve as reservoirs for bacteria with human pathogenic potential. Diagnosis of infection includes direct microscopy of wet mounts of cerebrospinal fluid or stained smears of cerebrospinal fluid sediment, light or electron microscopy of tissues, in vitro cultivation of Acanthamoeba, and histological assessment of frozen or paraffin-embedded sections of brain or cutaneous lesion biopsy material. Immunocytochemistry, chemifluorescent dye staining, PCR, and analysis of DNA sequence variation also have been employed for laboratory diagnosis. Treatment of Acanthamoeba infections has met with mixed results. However, chlorhexidine gluconate, alone or in combination with propamidene isethionate, is effective in some patients. Furthermore, effective treatment is complicated since patients may present with underlying disease and Acanthamoeba infection may not be recognized. Since an increase in the number of cases of Acanthamoeba infections has occurred worldwide, these protozoa have become increasingly important as agents of human disease.

  1. Protein profiles and immunoreactivities of Acanthamoeba morphological groups and genotypes.

    PubMed

    Pumidonming, Wilawan; Koehsler, Martina; Leitsch, David; Walochnik, Julia

    2014-11-01

    Acanthamoeba is a free-living protozoan found in a wide variety of habitats. A classification of Acanthamoeba into currently eighteen genotypes (T1-T18) has been established, however, data on differences between genotypes on the protein level are scarce. The aim of this study was to compare protein and immunoreactivity profiles of Acanthamoeba genotypes. Thirteen strains, both clinical and non-clinical, from genotypes T4, T5, T6, T7, T9, T11 and T12, representing three morphological groups, were investigated for their protein profiles and IgG, IgM and IgA immunoreactivities. It was shown that protein and immunoreactivity profiles of Acanthamoeba genotypes T4, T5, T6, T7, T9, T11 and T12 are clearly distinct from each other, but the banding patterns correlate to the morphological groups. Normal human sera revealed anti-Acanthamoeba antibodies against isolates of all investigated genotypes, interestingly, however only very weak IgM and virtually no IgA immunoreactivity with T7 and T9, both representing morphological group I. The strongest IgG, IgM and IgA immunoreactivities were observed for genotypes T4, T5 and T6. Differences of both, protein and immunological patterns, between cytopathic and non-cytopathic strains, particularly within genotype T4, were not at the level of banding patterns, but rather in expression levels.

  2. Expressed sequence tags (ESTs) analysis of Acanthamoeba healyi

    PubMed Central

    Kong, Hyun-Hee; Hwang, Mee-Yeul; Kim, Hyo-Kyung

    2001-01-01

    Randomly selected 435 clones from Acanthamoeba healyi cDNA library were sequenced and a total of 387 expressed sequence tags (ESTs) had been generated. Based on the results of BLAST search, 130 clones (34.4%) were identified as the genes enconding surface proteins, enzymes for DNA, energy production or other metabolism, kinases and phosphatases, protease, proteins for signal transduction, structural and cytoskeletal proteins, cell cycle related proteins, transcription factors, transcription and translational machineries, and transporter proteins. Most of the genes (88.5%) are newly identified in the genus Acanthamoeba. Although 15 clones matched the genes of Acanthamoeba located in the public databases, twelve clones were actin gene which was the most frequently expressed gene in this study. These ESTs of Acanthamoeba would give valuable information to study the organism as a model system for biological investigations such as cytoskeleton or cell movement, signal transduction, transcriptional and translational regulations. These results would also provide clues to elucidate factors for pathogenesis in human granulomatous amoebic encephalitis or keratitis by Acanthamoeba. PMID:11441502

  3. [Treatment of Acanthamoeba keratitis: possibilities, problems, and new approaches].

    PubMed

    Walochnik, Julia; Duchêne, Michael; Eibl, Hansjörg; Aspöck, Horst

    2003-01-01

    Acanthamoeba keratitis is a corneal disease associated predominantly with contact lens wear. The occurrence of Acanthamoeba keratitis has been rising since 1990 in correlation to the growing number of contact lens wearers. To date approximately 2000 cases have been published around the world. Due to the complicated diagnostics, the elaborate treatment and the usually bad compliance of the patients, Acanthamoeba keratitis unfortunately very often takes a serious progression, which may lead to serious visual loss and perforating keratoplasty. Today, local treatment with a combination of polyhexamethylene biguanide (PHMB) and propamidine isethionate (Brolene) is considered the first line therapy for Acanthamoeba keratitis. Alternatively also a combination of propamidine and chlorhexidine or neomycine achieves good therapeutic results. However, the complicated mode of application consistently remains a problem. The intensive local treatment, i.e. hourly application of therapeutics during the first three days day and night makes hospitalization inevitable. Moreover, sufficient efficacy can not always be achieved, and also resistance against propamidine has already been observed. Recently propamidine has sometimes been replaced by hexamidine, which seems to have a greater cysticidal activity. A new path might be struck by the application of alkylphosphocholines. These are phosphocholines esterified to aliphatic alcohols. They exhibit in vitro and in vivo antineoplastic activity and have been shown to be cytotoxic against Leishmania donovani, Trypanosoma cruzi, and Entamoeba histolytica. A recent study has demonstrated that particularly hexadecylphosphocholine is highly effective also against various strains of Acanthamoeba.

  4. Recent advances in the treatment of Acanthamoeba keratitis.

    PubMed

    Kumar, Raman; Lloyd, David

    2002-08-15

    Infection of the eye caused by Acanthamoeba species constitutes a burgeoning and unsolved problem. Of individuals with Acanthamoeba keratitis, 85% wear contact lenses; abrasion of the cornea is implicated. Corneal infection often can be prevented by good lens care and hygiene. Severe Acanthamoeba keratitis often can be very difficult to treat; surgery can be less than successful and may lead to further problems. The encysted stage in the life cycle of Acanthamoeba species appears to cause the most problems; many biocides are ineffective in killing the highly resistant cysts. Combination therapy--that is, use of 2 or 3 biocides, sometimes with antibacterial antibiotics--appears to work best. Recurrence is common if treatment is stopped prematurely. Immunologic methods are being investigated as a form of prevention, and oral immunization of animals recently has been successful in the prevention of Acanthamoeba keratitis by inducing immunity before infection occurs. Immunization thus may eventually become the best approach for reduction of the incidence of amebic infection in humans.

  5. Acanthamoeba keratitis: an emerging disease gathering importance worldwide?

    PubMed

    Lorenzo-Morales, Jacob; Martín-Navarro, Carmen María; López-Arencibia, Atteneri; Arnalich-Montiel, Francisco; Piñero, José E; Valladares, Basilio

    2013-04-01

    Acanthamoeba keratitis (AK) is increasingly being recognized as a severe sight-threatening ocular infection worldwide. Although contact lens wear is the leading risk factor for AK, Acanthamoeba parasites are also an important cause of keratitis in non-contact lens wearers. Diagnosis of AK is challenging, and the available treatments are lengthy and not fully effective against all strains. The pathogenesis of Acanthamoeba is still under study, and the identification of the key factors involved in this process should be useful for the development of fully effective therapies. This review focuses on recent developments on AK pathogenesis and diagnosis as well as novel strategies for the evaluation of anti-amoebic agents that could be applied in the near future against these pathogens.

  6. Acanthamoeba T4 genotype associated with keratitis infections in Tunisia.

    PubMed

    Dendana, F; Sellami, H; Trabelsi, H; Neji, S; Cheikhrouhou, F; Makni, F; Ayadi, A

    2013-01-01

    Acanthamoeba keratitis (AK) is a sight-threatening infection. We report five cases of AK diagnosed from 2005 to 2009 in the Laboratory of Parasitology-Mycology at Habib Bourguiba Sfax Hospital, Tunisia. All were associated with improper care of contact lenses (rinsing of contact lenses with tap water and inappropriate cleaning) and lens storage. The patients displayed different clinical presentations: corneal inflammation, corneal ulceration, and corneal abscess. The diagnosis was made after direct examination, culture, and polymerase chain reaction amplification with specific primers. The genotype classification was based on the highly variable DF3 region in the 18S rRNA gene. This is the first study characterizing Acanthamoeba genotype in Tunisia and North Africa. All Acanthamoeba isolates were associated to the T4 genotype. Three different DF3 sequence types were related to AK infections T4/10, T4/15, and T4/16.

  7. IL-17A-mediated protection against Acanthamoeba keratitis.

    PubMed

    Suryawanshi, Amol; Cao, Zhiyi; Sampson, James F; Panjwani, Noorjahan

    2015-01-15

    Acanthamoeba keratitis (AK) is a very painful and vision-impairing infection of the cornea that is difficult to treat. Although past studies have indicated a critical role of neutrophils and macrophages in AK, the relative contribution of the proinflammatory cytokine, IL-17A, that is essential for migration, activation, and function of these cells into the cornea is poorly defined. Moreover, the role of the adaptive immune response, particularly the contribution of CD4(+) T cell subsets, Th17 and regulatory T cells , in AK is yet to be understood. In this report, using a mouse corneal intrastromal injection-induced AK model, we show that Acanthamoeba infection induces a strong CD4(+) T effector and regulatory T cell response in the cornea and local draining lymph nodes. We also demonstrate that corneal Acanthamoeba infection induces IL-17A expression and that IL-17A is critical for host protection against severe AK pathology. Accordingly, IL-17A neutralization in Acanthamoeba-infected wild-type mice or Acanthamoeba infection of mice lacking IL-17A resulted in a significantly increased corneal AK pathology, increased migration of inflammatory cells at the site of inflammation, and a significant increase in the effector CD4(+) T cell response in draining lymph nodes. Thus, in sharp contrast with other corneal infections such as herpes and Pseudomonas aeruginosa keratitis where IL-17A exacerbates corneal pathology and inflammation, the findings presented in this article suggest that IL-17A production after Acanthamoeba infection plays an important role in host protection against invading parasites.

  8. The association of contact lens solution use and Acanthamoeba keratitis

    PubMed Central

    Joslin, Charlotte E.; Tu, Elmer Y.; Shoff, Megan E.; Booton, Gregory C.; Fuerst, Paul A.; McMahon, Timothy T.; Anderson, Robert J.; Dworkin, Mark S.; Sugar, Joel; Davis, Faith G.; Stayner, Leslie T.

    2009-01-01

    Purpose Diagnosis of Acanthamoeba keratitis, a rare but serious corneal infection, has recently increased significantly at the University of Illinois at Chicago (UIC) Cornea Service. The purpose is to investigate Acanthamoeba keratitis risk factors. Design Retrospective case-control study. Methods Setting University, tertiary care hospital. Patients Fifty-five Acanthamoeba keratitis cases with contact lens use were diagnosed between May 1, 2003 and September 15, 2006. Clinic-matched controls with contact lens use were recruited. Subjects completed surveys targeting lens hygiene, contact lens solution use, and water exposure. Main Outcome Measure Acanthamoeba keratitis. Results Thirty-nine (73.6%) cases and 113 (65.3%) controls participated; 38 cases had complete contact lens data. Thirty-five of 38 cases (92.1%) and 47 of 100 controls (47.0%) used soft lenses. Analysis was performed on 30 cases and 39 controls with matched pairs with soft lens use. Exclusive use of AMO Complete MoisturePlus Multi-Purpose Solution was independently associated with Acanthamoeba keratitis in multivariable analysis (55.2% vs. 10.5%; OR, 16.67; 95% CI, 2.11–162.63; p = 0.008). However, 38.8% of cases reported no use of AMO Complete MoisturePlus Multi-Purpose Solution or used it in combination with other solutions. Although not statistically significant, additional hygiene-related variables (solution ‘reuse’, lack of ‘rubbing’, and showering with lenses) suggest a pattern of risk,. Conclusions AMO Complete MoisturePlus Multi-Purpose Solution use is independently associated with Acanthamoeba keratitis among soft contact lens users. However, it does not explain all cases, suggesting additional factors. Further research into environmental risk factors and hygiene practices is warranted, especially considering this is the second outbreak of an atypical, contact lens-related infection. PMID:17588524

  9. IL-17A-Mediated Protection against Acanthamoeba Keratitis

    PubMed Central

    Suryawanshi, Amol; Cao, Zhiyi; Sampson, James F.

    2014-01-01

    Acanthamoeba keratitis (AK) is a very painful and vision impairing infection of the cornea that is difficult to treat. Although past studies have indicated a critical role of neutrophils and macrophages in AK, the relative contribution of the proinflammatory cytokine, IL-17A, that is essential for migration, activation and function of these cells into the cornea is poorly defined. Moreover, the role of the adaptive immune response, particularly the contribution of CD4+ T cell subsets, Th17 and Treg cells, in AK is yet to be understood. In this report, using a mouse corneal intrastromal injection-induced AK model, we show that Acanthamoeba infection induces a strong CD4+ T effector and regulatory T cell response in the cornea as well as local draining lymph nodes (dLN). We also demonstrate that corneal Acanthamoeba infection induces IL-17A expression and that IL-17A is critical for host protection against severe AK pathology. Accordingly, IL-17A neutralization in Acanthamoeba-infected wild-type mice or Acanthamoeba infection of mice lacking IL-17A resulted in a significantly increased corneal AK pathology, increased migration of inflammatory cells at the site of inflammation and a significant increase in the effector CD4+ T cell response in dLN. Thus, in sharp contrast to other corneal infections such as herpes and P. aeruginosa keratitis where IL-17A exacerbates corneal pathology and inflammation, findings presented in this manuscript suggest that IL-17A production after Acanthamoeba infection plays an important role in host protection against invading parasites. PMID:25505284

  10. Acanthamoeba DNA can be directly amplified from corneal scrapings.

    PubMed

    El-Sayed, Nagwa Mostafa; Younis, Mohamed Saad; Elhamshary, Azza Mohamed; Abd-Elmaboud, Amina Ibrahim; Kishik, Shereen Magdy

    2014-09-01

    This study evaluated the performance of direct amplification of Acanthamoeba-DNA bypassing DNA extraction in the diagnosis of Acanthamoeba keratitis in clinically suspected cases in comparison to direct microscopic examination and in vitro culture. Corneal scrapings were collected from 110 patients who were clinically suspected to have Acanthamoeba keratitis, 63 contact lens wearers (CLW), and 47 non-contact lens wearers (NCLW). Taken samples were subjected to direct microscopic examination, cultivation onto the non-nutrient agar plate surface seeded with Escherichia coli, and PCR amplification. The diagnostic performance of these methods was statistically compared. The results showed that Acanthamoeba infection was detected in 21 (19.1%) of clinically suspected cases (110); 17 (81%) of them were CLW and the remaining 4 (19%) positive cases were NCLW. Regarding the used diagnostic methods, it was found that direct amplification of Acanthamoeba DNA bypassing nucleic acid extraction was superior to microscopy and culture in which 21 cases (19.1%) were positive for Acanthamoeba by PCR compared to 19 positive cases by culture (17.3%) and one case (0.9%) by direct smear. The difference in detection rates between culture and direct smear was highly statistically significant (P = 0.001). On the other hand, there was no significant difference in detection rates between culture and PCR (P = 0.86). On using culture as the gold standard, PCR showed three false-positive samples that were negative by culture and one false-negative sample that was positive by culture. At the same time, direct smear showed 18 false-negative samples. The sensitivity, specificity, positive predictive value, negative predictive value, and diagnostic accuracy of PCR were 94.7, 96.7, 85.7, 98.9, and 96.4, respectively, while those of direct smear were 5.3, 100, 100, 83.5, and 83.6, respectively. In conclusion, direct amplification of Acanthamoeba-DNA bypassing DNA extraction is a reliable

  11. Survival of Environmental Mycobacteria in Acanthamoeba polyphaga

    PubMed Central

    Adékambi, Toïdi; Ben Salah, Skandar; Khlif, Mohamed; Raoult, Didier; Drancourt, Michel

    2006-01-01

    Free-living amoebae in water are hosts to many bacterial species living in such an environment. Such an association enables bacteria to select virulence factors and survive in adverse conditions. Waterborne mycobacteria (WBM) are important sources of community- and hospital-acquired outbreaks of nontuberculosis mycobacterial infections. However, the interactions between WBM and free-living amoebae in water have been demonstrated for only few Mycobacterium spp. We investigated the ability of a number (n = 26) of Mycobacterium spp. to survive in the trophozoites and cysts of Acanthamoeba polyphaga. All the species tested entered the trophozoites of A. polyphaga and survived at this location over a period of 5 days. Moreover, all Mycobacterium spp. survived inside cysts for a period of 15 days. Intracellular Mycobacterium spp. within amoeba cysts survived when exposed to free chlorine (15 mg/liter) for 24 h. These data document the interactions between free-living amoebae and the majority of waterborne Mycobacterium spp. Further studies are required to examine the effects of various germicidal agents on the survival of WBM in an aquatic environment. PMID:16957218

  12. Optical-vortex pair creation and annihilation and helical astigmatism of a nonplanar ring resonator

    SciTech Connect

    Heckenberg, N.R.; Vaupel, M.; Malos, J.T.; Weiss, C.O.

    1996-09-01

    The creation and annihilation of pairs of optical vortices have been studied in transitions between patterns produced in a photorefractive oscillator. Smooth metamorphosis between stable patterns occurs through pair creation or annihilation but can be modeled using superposition of modes taking into account lifting of degeneracy of helical modes by helical astigmatism of the resonator. {copyright} {ital 1996 The American Physical Society.}

  13. [Post-operative residual astigmatism after cataract surgery: Current surgical methods of treatment].

    PubMed

    Pisella, P-J

    2012-03-01

    Residual astigmatism after cataract surgery can be corrected by three different techniques: classic limbal relaxing incisions, easy to perform but with limited precision; laser refractive surgery (PRK or Lasik), additionally allowing for correction of spherical equivalent; and more recently the use of a piggyback toric intraocular lens in the ciliary sulcus.

  14. Refractive and Quality of Vision Outcomes with Toric IOL Implantation in Low Astigmatism

    PubMed Central

    Patrão, Lia Florim; de Moraes, Haroldo Vieira

    2016-01-01

    Purpose. To evaluate the refractive and the quality of vision outcomes of toric IOL implantation in patients with low astigmatism. Design. Prospective study of single-arm. Methods. Patients with corneal astigmatism range from 0,75 D to 1,5 D and cataract that underwent cataract surgery with toric IOL. The measurements were performed preoperatively and 6 weeks after the surgery. Patients were evaluated for visual acuity with and without correction, contrast sensitivity, static and dynamic refraction, and quality of life questionnaire. Pre- and postoperative values were compared and their variations were evaluated for linear correlation. Results. 21 eyes of 21 patients. Postoperative mean uncorrected visual acuity was 0.80 ± 0.19, and the best corrected visual acuity was 0.97 ± 0.15. p < 0.001 compared to preoperative values. The average postoperative refractive cylinder was −0.34 ± 0.39. The questionnaire's total value before and after surgery was, respectively, 43.20 ± 15.76 and 79.70 ± 10.11 (p < 0.001). The correlation coefficients between the values of the questionnaire variation and the UCVA, BCVA, and CS variation were, respectively, 0.548 (p = 0.005), 0.508 (p = 0.009), and 0.409 (p = 0.033). Conclusion. Patients with low astigmatism who underwent phacoemulsification with toric IOL implantation experienced significant decrease in refractive astigmatism and improvement in their quality of life. PMID:28070415

  15. Normalisation of asymmetric astigmatism after intralesional steroid injection for upper eye lid hemangioma in childhood.

    PubMed

    Langmann, A; Lindner, S

    1994-01-01

    Infantile hemangiomas affect about 5% (3%-8%) of the population, showing a predilection for the face. After a phase of rapid enlargement between the 3rd and the 9th month of life, 70% regress by the age of six after a period of stability. 43%-60% of the children with eye lid hemangiomas develop strabismic, anisometropic, or deprivation amblyopia. Previous studies found the majority of cases resulting from anisometropia (especially asymmetric astigmatism) rather than strabism or occlusion of the visual axis. Several methods of treatment--surgical excision, irradiation, sclerosing agents, systemic steroids, ligation, cryotherapy--have been used but all with a risk of local or systemic complications. Local injections of steroids are a simple method of therapy with a high rate of resolution of hemangiomas, but still with a high degree of bad visual output because of persistent astigmatism. In four children with asymmetric astigmatism (axis of astigmatism towards the hemangioma) in which the injection was given at the beginning of the phase of enlargement, amblyopia could be avoided by preventing corneal steepening from becoming permanent.

  16. Prevalence of Corneal Astigmatism and Anterior Segmental Biometry Characteristics Before Surgery in Chinese Congenital Cataract Patients

    PubMed Central

    Lin, Duoru; Chen, Jingjing; Liu, Zhenzhen; Wu, Xiaohang; Long, Erping; Luo, Lixia; Lin, Zhuoling; Li, Xiaoyan; Zhang, Li; Chen, Hui; Liu, Jinchao; Chen, Weirong; Lin, Haotian; Liu, Yizhi

    2016-01-01

    The prevalence and the distribution characteristics of corneal astigmatism (CA) and anterior segment biometry before surgery in Chinese congenital cataract (CC) patients are not completely understood. This study involved 400 CC patients from the Zhongshan Ophthalmic Center enrolled from February 2011 to August 2015. Data on CA, keratometry, central corneal thickness (CCT) and anterior chamber depth (ACD) were measured by the Pentacam Scheimpflug System. The mean age of patients was 54.27 months, and the ratio of boys to girls was 1.53:1. The mean CA was 2.03 diopters (D), and 39.25% of subjects had CA values ≥2 D. The most frequent (71.8%) diagnosis was with-the-rule astigmatism. Oblique astigmatism was present in 16.2% of cases, and 12% of cases had against-the-rule astigmatism. The mean keratometry measurement of cataractous eyes in bilateral patients was significantly larger than that in unilateral patients. Girls had a larger mean keratometry but a thinner CCT than did boys. The CA, CCT, and ACD of cataractous eyes were significantly larger than those of non-cataractous eyes in unilateral patients. The CA, mean keratometry, CCT, and ACD in CC patients varied with age, gender, and laterality. Fully understanding these characteristics may help inform guidelines and treatment decisions in CC patients. PMID:26912400

  17. Pseudophakic astigmatism reduction with femtosecond laser-assisted corneal arcuate incisions: a pilot study

    PubMed Central

    Blehm, Clayton; Potvin, Richard

    2017-01-01

    Purpose The aim of this study was to assess the effectiveness of the Verion-LenSx guided arcuate incision technique to reduce refractive astigmatism in a pseudophakic population. Patients and methods A prospective single-arm study was conducted in which one or both eyes of subjects required reduction of 1.0–2.0 D of refractive astigmatism after previous cataract surgery or refractive lens exchange. The surgeon used the refractive cylinder in the eye and the Woodcock astigmatism nomogram for preoperative planning, while the LenSx femtosecond laser with the Verion Image Guided System was used to create all arcuate incisions. The primary outcome measure was the uncorrected monocular distance visual acuity (UCVA). Secondary outcome measures included the change in corneal astigmatism, the change in refractive astigmatism, the best-corrected visual acuity and spectacle independence at distance from preoperative stage to 1 month and 2 months postoperatively. Results Twenty-eight eyes of 18 subjects were treated. The best-corrected visual acuity at the 2-month postoperative (PO) stage was not statistically significantly different from the preoperative visual acuity (0.02 logarithm of the minimum angle of resolution [logMAR] in both cases, P>0.05). Uncorrected visual acuity was statistically significantly better at the 2-month PO stage relative to the preoperative value (0.14 versus 0.34 logMAR, P<0.01). The mean change in refractive cylinder from the preoperative stage to the 2-month PO stage was 1.0 D. At the 2-month PO stage, two-thirds of the subjects (12/18) reported that they did not use glasses for distance vision and that their spectacle use for distance vision at 2 months was “lower” or “much lower” than the preoperative stage; in 71% of eyes (20/28), the residual refractive cylinder was ≤0.50 D. Vector changes in keratometric astigmatism were weakly associated with changes in refractive cylinder. Conclusion Arcuate incisions made with a femtosecond

  18. The role of sideport incision in astigmatism change after cataract surgery

    PubMed Central

    Theodoulidou, Sofia; Asproudis, Ioannis; Kalogeropoulos, Christos; Athanasiadis, Aristidis; Aspiotis, Miltiadis

    2015-01-01

    Purpose To study the changes in corneal astigmatism after cataract surgery when the sideport incision is performed at a predetermined location away from the tunnel incision. Setting General Hospital of Piraeus “Tzaneio”, Attiki, Greece. Materials and methods A total of 333 eyes with corneal astigmatism ≤1.5 diopters (D) underwent cataract surgery. A three-step superotemporal clear corneal incision for the right eye and a superonasal clear corneal incision for the left eye (3.0 mm) was made, while the sideport incision was located at <90°, 90°–110°, and >110°. Keratometric data were measured with corneal topography EyeSys Vista 2000 pre- and postoperatively at the 1st and 6th month. Surgically induced astigmatism was calculated by vector analysis. We noted all cases in which a change >0.5 D in corneal astigmatic power occurred, as well as a change >20° in axis torque, despite axis direction. Results After multiple logistic regression analysis was conducted, cases with >110° distance between the tunnel and sideport incision had 2.22 times (P=0.021) greater likelihood for having changed >0.5 D in astigmatic power at the 1st month and 3.45 times (P=0.031) at the 6th month postoperatively, as compared with cases with a 90°–110° distance between the tunnel and sideport incision. As for the change in the astigmatic axis, cases with <90° distance had a 4.18 times greater likelihood for having a change >20° (P<0.001) (preoperative to 1st month) as compared with cases having 90°–110° of distance. Conclusion For surgeons that operate only from the superior position, we propose that in order to produce an incision that is as “astigmatically neutral” as possible, they should perform the sideport incision at a 90°–110° distance. PMID:26346741

  19. [Results of corneal and total astigmatism estimation by different methods in myopic patients wearing orthokeratology contact lenses].

    PubMed

    Tarutta, E P; Aliaeva, O O; Verzhanskaia, T Iu; Milash, S V

    2013-01-01

    Reports have been made that corneal aberrations of all orders, including astigmatism, often significantly increase with the use of night orthokeratology lenses. In this study the dynamic changes of total and corneal astigmatism in myopes using orthokeratology lenses was evaluated by different methods. The study enrolled 38 patients (76 eyes) with low and medium myopia (28 and 48 eyes correspondingly) and initial astigmatism less than 2 diopters. The assessment was made before and in different terms after the patient started to wear orthokeratology lenses. Induced astigmatism (> or =1 diopter) was found in more than 50% of cases. The degree of astigmatism gradually increased from the centre to the periphery within the papillary zone. The maximum values were found within a 4-mm zone ("uptake zone") and minimal - within a 8-mm zone ("equalization zone"). In all patients, despite the presence of induced astigmatism and residual myopia (0.83+/-0.09 diopters in average), distance visual acuity was high enough without an additional correction (0.82+/-0.05 in average). Apparently, in these patients the aberrations (astigmatism in particular) exceed the focal depth.

  20. Epidemiological typing of Acanthamoeba strains isolated from keratitis cases in Belgium.

    PubMed

    De Jonckheere, J F

    2003-01-01

    From the corneas of nine keratitis patients and from their contact lenses, contact lens boxes and saline solutions, 15 strains of Acanthamoeba have been isolated. An Acanthamoeba strain was isolated from the swimming pool where one of the patients swam, while in the tapwater of the houses of three patients investigated, no Acanthamoeba could be detected. All the Acanthamoeba isolates from the cornea belong to genotype T4, but are different subtypes of T4. The Acanthamoeba detected on the contact lenses (and/or associated paraphernalia) of a patient are of the same subtype as that isolated from the cornea. The only Acanthamoeba strain isolated from a contact lens which was not related to an Acanthamoeba keratitis infection proved to be another genotype. A strain of Hartmannella from a cornea and two vahlkampfiids isolated from contact lenses had no connection with keratitis. This study confirms that, as found elsewhere, only Acanthamoeba genotype T4 of the 12 known Acanthamoeba genotypes is responsible for keratitis in Belgium. Most cases of Acanthamoeba keratitis cases are due to poor hygiene in the treatment (cleaning and storage) of contact lenses.

  1. Surgically-induced astigmatism in combined ECCE with filtering procedures compared to ECCE alone.

    PubMed

    Choplin, N T; Monroe, J F

    1992-02-01

    Extracapsular cataract extraction with implantation of a posterior chamber lens combined with filtering surgery (glaucoma triple procedure) is frequently employed in the co-management of cataract and glaucoma. Nineteen triple procedures with a minimum of six months follow up were compared to 19 cases of extracapsular cataract extraction with lens implant matched for age, sex, and surgeon with regard to surgically induced astigmatism as determined by vector analysis. Follow up averaged 10 months for the triple group and 14 months for the controls. There was no statistically significant difference in preoperative astigmatism between the two groups nor in the mean number of sutures cut. Postoperatively, the keratometric cylinder averaged 2.55 +/- 1.54 diopters for the cases and 1.20 +/- 1.11 D for the controls; the difference of 1.36 D was statistically significant (P = .004). The postoperative refractive cylinder was 2.34 +/- 1.54 D for the cases and 1.29 +/- 1.07 D for the controls; the difference of 1.05 D was statistically significant (P = .017). With regard to surgically-induced astigmatism, vector analysis showed that the cases averaged 2.18 +/- 1.25 D and the controls 1.23 +/- 0.81 D; the difference of 0.95 D was statistically significant (P = .006). With regard to surgically-induced astigmatism, vector analysis showed that the cases averaged 2.18 +/- 1.25 D and the controls 1.23 +/- 0.81 D; the difference of 0.95 D was statistically significant (P = .006). When analyzed for change along the vertical meridian, the cases averaged 1.12 +/- 1.57 D surgically-induced against-the-rule astigmatism, as compared with 0.30 +/- 1.16 D for the controls (difference, 0.83 D; P = .062). The glaucoma triple procedures induced approximately 1.00 D more cylinder than the controls.

  2. Acanthamoeba polyphaga mimivirus virophage seroconversion in travelers returning from Laos.

    PubMed

    Parola, Philippe; Renvoisé, Aurélie; Botelho-Nevers, Elisabeth; La Scola, Bernard; Desnues, Christelle; Raoult, Didier

    2012-09-01

    During January 2010, a husband and wife returned from Laos to France with probable parasitic disease. Increased antibodies against an Acanthamoeba polyphaga mimivirus virophage indicated seroconversion. While in Laos, they had eaten raw fish, a potential source of the virophage. This virophage, associated with giant viruses suspected to cause pneumonia, could be an emerging pathogen.

  3. Colonization of broilers by Campylobacter jejuni internalized within Acanthamoeba castellanii

    Technology Transfer Automated Retrieval System (TEKTRAN)

    We present the first report that Campylobacter jejuni, internalized within Acanthamoeba castellanii, colonized broilers. After 1, 3, 7 and 14 days post challenge none of the broilers challenged with negative controls were colonized, but were with internalized C. jejuni. The biology of protozoa-Cam...

  4. Cytotoxic Activities of Alkylphosphocholines against Clinical Isolates of Acanthamoeba spp.

    PubMed Central

    Walochnik, Julia; Duchêne, Michael; Seifert, Karin; Obwaller, Andreas; Hottkowitz, Thomas; Wiedermann, Gerhard; Eibl, Hansjörg; Aspöck, Horst

    2002-01-01

    Free-living amoebae of the genus Acanthamoeba are causing serious chronic conditions such as destructive keratitis in contact lens wearers or granulomatous amoebic encephalitis in individuals with compromised immune systems. Both are characterized by the lack of availability of sufficiently effective and uncomplicated, manageable treatments. Hexadecylphosphocholine (miltefosine) is licensed for use as a topical antineoplastic agent, but it is also active in vitro against several protozoan parasites, and it was applied very successfully for the treatment of human visceral leishmaniasis. The aim of our study was to evaluate the efficacy of hexadecylphosphocholine and other alkylphosphocholines (APCs) against Acanthamoeba spp. The in vitro activities of eight different APCs against three Acanthamoeba strains of various pathogenicities were determined. All substances showed at least amoebostatic effects, and some of them disrupted the amoebae, as shown by the release of cytoplasmic enzyme activity. Hexadecylphosphocholine exhibited the highest degree of cytotoxicity against trophozoites, resulting in complete cell death at a concentration as low as 40 μM, and also displayed significant cysticidal activity. Hexadecylphosphocholine may be a promising new candidate for the topical treatment of Acanthamoeba keratitis and, conceivably, even for the oral treatment of granulomatous amoebic encephalitis. PMID:11850250

  5. Cytotoxic activities of alkylphosphocholines against clinical isolates of Acanthamoeba spp.

    PubMed

    Walochnik, Julia; Duchêne, Michael; Seifert, Karin; Obwaller, Andreas; Hottkowitz, Thomas; Wiedermann, Gerhard; Eibl, Hansjörg; Aspöck, Horst

    2002-03-01

    Free-living amoebae of the genus Acanthamoeba are causing serious chronic conditions such as destructive keratitis in contact lens wearers or granulomatous amoebic encephalitis in individuals with compromised immune systems. Both are characterized by the lack of availability of sufficiently effective and uncomplicated, manageable treatments. Hexadecylphosphocholine (miltefosine) is licensed for use as a topical antineoplastic agent, but it is also active in vitro against several protozoan parasites, and it was applied very successfully for the treatment of human visceral leishmaniasis. The aim of our study was to evaluate the efficacy of hexadecylphosphocholine and other alkylphosphocholines (APCs) against Acanthamoeba spp. The in vitro activities of eight different APCs against three Acanthamoeba strains of various pathogenicities were determined. All substances showed at least amoebostatic effects, and some of them disrupted the amoebae, as shown by the release of cytoplasmic enzyme activity. Hexadecylphosphocholine exhibited the highest degree of cytotoxicity against trophozoites, resulting in complete cell death at a concentration as low as 40 microM, and also displayed significant cysticidal activity. Hexadecylphosphocholine may be a promising new candidate for the topical treatment of Acanthamoeba keratitis and, conceivably, even for the oral treatment of granulomatous amoebic encephalitis.

  6. Identification of Unusual Phospholipid Fatty Acyl Compositions of Acanthamoeba castellanii

    PubMed Central

    Palusinska-Szysz, Marta; Kania, Magdalena; Turska-Szewczuk, Anna; Danikiewicz, Witold; Russa, Ryszard; Fuchs, Beate

    2014-01-01

    Acanthamoeba are opportunistic protozoan pathogens that may lead to sight-threatening keratitis and fatal granulomatous encephalitis. The successful prognosis requires early diagnosis and differentiation of pathogenic Acanthamoeba followed by aggressive treatment regimen. The plasma membrane of Acanthamoeba consists of 25% phospholipids (PL). The presence of C20 and, recently reported, 28- and 30-carbon fatty acyl residues is characteristic of amoeba PL. A detailed knowledge about this unusual PL composition could help to differentiate Acanthamoeba from other parasites, e.g. bacteria and develop more efficient treatment strategies. Therefore, the detailed PL composition of Acanthamoeba castellanii was investigated by 31P nuclear magnetic resonance spectroscopy, thin-layer chromatography, gas chromatography, high performance liquid chromatography and liquid chromatography-mass spectrometry. Normal and reversed phase liquid chromatography coupled with mass spectrometric detection was used for detailed characterization of the fatty acyl composition of each detected PL. The most abundant fatty acyl residues in each PL class were octadecanoyl (18∶0), octadecenoyl (18∶1 Δ9) and hexadecanoyl (16∶0). However, some selected PLs contained also very long fatty acyl chains: the presence of 28- and 30-carbon fatty acyl residues was confirmed in phosphatidylethanolamine (PE), phosphatidylserine, phosphatidic acid and cardiolipin. The majority of these fatty acyl residues were also identified in PE that resulted in the following composition: 28∶1/20∶2, 30∶2/18∶1, 28∶0/20∶2, 30∶2/20∶4 and 30∶3/20∶3. The PL of amoebae are significantly different in comparison to other cells: we describe here for the first time unusual, very long chain fatty acids with Δ5-unsaturation (30∶35,21,24) and 30∶221,24 localized exclusively in specific phospholipid classes of A. castellanii protozoa that could serve as specific biomarkers for the presence of these

  7. Bioassay guided isolation and identification of anti-Acanthamoeba compounds from Tunisian olive leaf extracts.

    PubMed

    Sifaoui, Ines; López-Arencibia, Atteneri; Ticona, Juan Carlos; Martín-Navarro, Carmen M; Reyes-Batlle, María; Mejri, Mondher; Lorenzo-Morales, Jacob; Jiménez, Antonio Ignacio; Valladares, Basilio; Lopez-Bazzocchi, Isabel; Abderabba, Manef; Piñero, José E

    2014-11-01

    Pathogenic Acanthamoeba strains are causative agents of Granulomatous Amoebic Encephalitis (GAE) and Acanthamoeba keratitis (AK) worldwide. The existence of the cyst stage complicates Acanthamoeba therapy as it is highly resistant to antibiotics and physical agents. The aim of this study was to investigate the activity of Limouni olive leaf cultivar against the trophozoite stage of Acanthamoeba. The ethyl acetate and methanol extracts of this variety were tested against Acanthamoeba castellanii Neff. The ethyl acetate extract of olive leaf was the most active showing an IC50 of 5.11±0.71μg/ml of dry extract. Bio-guided fractionation of this extract was conducted and led to the identification of three active compounds namely oleanolic and maslinic acids and oleuropein which could be used for the development of novel therapeutic approaches against Acanthamoeba infections.

  8. Acanthamoeba can be differentiated by the polymerase chain reaction and simple plating assays.

    PubMed

    Khan, N A; Jarroll, E L; Paget, T A

    2001-09-01

    Acanthamoeba are opportunistic pathogens with invasive and noninvasive species. For clinical purposes it is important to differentiate potentially pathogenic from nonpathogenic isolates. For the rapid and sensitive identification of Acanthamoeba at the genus level, we used a polymerase chain reaction (PCR)-based method which detected as few as five cells. Further, we tested nine isolates of Acanthamoeba for their ability to produce cytopathic effects (CPE) on corneal epithelial cells. On the basis of the results, Acanthamoeba were divided into pathogenic or nonpathogenic groups. However, because CPE assays are not available to every diagnostic laboratory, we developed a simple plating assay based on osmotolerance which correlated well with the CPE assays. Pathogenic Acanthamoeba showed growth on higher osmolarity (agar plates containing one molar mannitol), while growth of nonpathogens was inhibited on these plates. In conclusion, we have developed methods for the rapid identification and differentiation of Acanthamoeba.

  9. Extended depth of focus intra-ocular lens: a solution for presbyopia and astigmatism

    NASA Astrophysics Data System (ADS)

    Zlotnik, Alex; Raveh, Ido; Ben Yaish, Shai; Yehezkel, Oren; Belkin, Michael; Zalevsky, Zeev

    2010-02-01

    Purpose: Subjects after cataract removal and intra-ocular lens (IOL) implantation lose their accommodation capability and are left with a monofocal visual system. The IOL refraction and the precision of the surgery determine the focal distance and amount of astigmatic aberrations. We present a design, simulations and experimental bench testing of a novel, non-diffractive, non-multifocal, extended depth of focus (EDOF) technology incorporated into an IOL that allows the subject to have astigmatic and chromatic aberrations-free continuous focusing ability from 35cm to infinity as well as increased tolerance to IOL decentration. Methods: The EDOF element was engraved on a surface of a monofocal rigid IOL as a series of shallow (less than one micron deep) concentric grooves around the optical axis. These grooves create an interference pattern extending the focus from a point to a length of about one mm providing a depth of focus of 3.00D (D stands for Diopters) with negligible loss of energy at any point of the focus while significantly reducing the astigmatic aberration of the eye and that generated during the IOL implantation. The EDOF IOL was tested on an optical bench simulating the eye model. In the experimental testing we have explored the characteristics of the obtained EDOF capability, the tolerance to astigmatic aberrations and decentration. Results: The performance of the proposed IOL was tested for pupil diameters of 2 to 5mm and for various spectral illuminations. The MTF charts demonstrate uniform performance of the lens for up to 3.00D at various illumination wavelengths and pupil diameters while preserving a continuous contrast of above 25% for spatial frequencies of up to 25 cycles/mm. Capability of correcting astigmatism of up to 1.00D was measured. Conclusions: The proposed EDOF IOL technology was tested by numerical simulations as well as experimentally characterized on an optical bench. The new lens is capable of solving presbyopia and astigmatism

  10. Thermotolerant Acanthamoeba spp. isolated from therapeutic hot springs in Northwestern Iran.

    PubMed

    Solgi, Rahmat; Niyyati, Maryam; Haghighi, Ali; Taghipour, Niloofar; Tabaei, Seyyed Javad Seyyed; Eftekhar, Mohamad; Nazemalhosseini Mojarad, Ehsan

    2012-12-01

    This study was conducted to address the distribution of Acanthamoeba genotypes in therapeutic hot springs in Iran. Sixty water and sediment samples were collected from bicarbonate, sulphur, and sodium chloride thermal springs in the northwest. All hot springs examined are used mainly for health purposes in Iran. Acanthamoeba were identified by both morphology and PCR (polymerase chain reaction). Genotype identification was based on the sequencing of a highly variable and informative region of Diagnostic Fragment 3 (stem 29-1 of 18S rRNA gene) within Acanthamoeba-specific amplimer (ASA.S1). Twenty percent of hot springs were contaminated with thermotolerant Acanthamoeba belonging to the potentially pathogenic T4 and T3 genotypes. A high number (91.7%) of strains showed growth at 37 °C, and eight isolates showed growth at 42 °C. A single isolate (HSNW2) was detected in waters at 70 °C. The presence of thermotolerant Acanthamoeba highlights a risk factor for susceptible individuals, as Acanthamoeba-related keratitis continues to rise in Iran. Periodic surveillance of thermal waters as well as improved filtration and disinfection is recommended to prevent disease related to pathogenic Acanthamoeba. This is the first comprehensive molecular study of Acanthamoeba genotypes in hot springs in Iran and the first to report the occurrence of the T3 genotype (corresponding to Acanthamoeba griffini) in thermal water sources in this country.

  11. Acanthamoeba protease activity promotes allergic airway inflammation via protease-activated receptor 2.

    PubMed

    Park, Mi Kyung; Cho, Min Kyoung; Kang, Shin Ae; Park, Hye-Kyung; Kim, Dong-Hee; Yu, Hak Sun

    2014-01-01

    Acanthamoeba is a free-living amoeba commonly present in the environment and often found in human airway cavities. Acanthamoeba possesses strong proteases that can elicit allergic airway inflammation. To our knowledge, the aeroallergenicity of Acanthamoeba has not been reported. We repeatedly inoculated mice with Acanthamoeba trophozoites or excretory-secretory (ES) proteins intra-nasally and evaluated symptoms and airway immune responses. Acanthamoeba trophozoites or ES proteins elicited immune responses in mice that resembled allergic airway inflammation. ES proteins had strong protease activity and activated the expression of several chemokine genes (CCL11, CCL17, CCL22, TSLP, and IL-25) in mouse lung epithelial cells. The serine protease inhibitor phenyl-methane-sulfonyl fluoride (PMSF) inhibited ES protein activity. ES proteins also stimulated dendritic cells and enhanced the differentiation of naive T cells into IL-4-secreting T cells. After repeated inoculation of the protease-activated receptor 2 knockout mouse with ES proteins, airway inflammation and Th2 immune responses were markedly reduced, but not to basal levels. Furthermore, asthma patients had higher Acanthamoeba-specific IgE titers than healthy controls and we found Acanthamoeba specific antigen from house dust in typical living room. Our findings suggest that Acanthamoeba elicits allergic airway symptoms in mice via a protease allergen. In addition, it is possible that Acanthamoeba may be one of the triggers human airway allergic disease.

  12. Seasonal distribution of potentially pathogenic Acanthamoeba species from drinking water reservoirs in Taiwan.

    PubMed

    Kao, Po-Min; Hsu, Bing-Mu; Hsu, Tsui-Kang; Liu, Jorn-Hon; Chang, Hsiang-Yu; Ji, Wen-Tsai; Tzeng, Kai-Jiun; Huang, Shih-Wei; Huang, Yu-Li

    2015-03-01

    In order to detect the presence/absence of Acanthamoeba along with geographical variations, water quality variations and seasonal change of Acanthamoeba in Taiwan was investigated by 18S ribosomal RNA (rRNA) gene TaqMan quantitative real-time PCR. Samples were collected quarterly at 19 drinking water reservoir sites from November 2012 to August 2013. Acanthamoeba was detected in 39.5 % (30/76) of the water sample, and the detection rate was 63.2 % (12/19) from samples collected in autumn. The average concentration of Acanthamoeba was 3.59 × 10(4) copies/L. For geographic distribution, the detection rate for Acanthamoeba at the northern region was higher than the central and southern regions in all seasons. Results of Spearman rank test revealed that heterotrophic plate count (HPC) had a negative correlation (R = -0.502), while dissolved oxygen (DO) had a positive correlation (R = 0.463) in summer. Significant differences were found only between the presence/absence of Acanthamoeba and HPC in summer (Mann-Whitney U test, P < 0.05). T2 and T4 genotypes of Acanthamoeba were identified, and T4 was the most commonly identified Acanthamoeba genotypes. The presence of Acanthamoeba in reservoirs presented a potential public health threat and should be further examined.

  13. Characterization of isolates of Acanthamoeba from the nasal mucosa and cutaneous lesions of dogs.

    PubMed

    Carlesso, A M; Mentz, M B; da Machado, M L S; Carvalho, A; Nunes, T E T; Maschio, V J; Rott, M B

    2014-06-01

    Acanthamoeba spp. are free-living amoebae that are ubiquitously distributed in the environment and can cause encephalomyelitis in animals and humans. The factors that contribute to Acanthamoeba infections include parasite biology, genetic diversity, environmental spread, and host susceptibility. The aim of the present study was to characterize isolates of Acanthamoeba from the nasal mucosa and cutaneous lesions of dogs in order to access the occurence and pathogenicity of these organisms in this animal group. We studied 13 isolates of Acanthamoeba confirmed by polymerase chain reaction. They were sequenced, the genotype was determined, and their potential of pathogenicity was evaluated.

  14. The role of the innate and adaptive immune responses in Acanthamoeba keratitis.

    PubMed

    Niederkorn, Jerry Y

    2002-01-01

    Infections of the corneal surface are an important cause of blindness. Protozoal, viral, bacterial, and helminthic infections of the cornea account for up to 9 million cases of corneal blindness. Free-living amoebae of the genus Acanthamoeba produce a progressive infection of the cornea called Acanthamoeba keratitis. Disease is usually transmitted by Acanthamoeba trophozoites bound to soft contact lenses. Infection of the cornea is initiated when the parasite binds to the corneal epithelial surface. Recrudescence can occur and suggests that the adaptive immune response is not aroused by corneal Acanthamoeba infections. Systemic immunization with Acanthamoeba antigens elicits robust Th1 cell-mediated immunity and serum IgG antibody, yet fails to prevent the development of Acanthamoeba keratitis. However, immunization via mucosal surfaces induces anti-Acanthamoeba IgA antibodies in the tears and provides solid protection against the development of Acanthamoeba keratitis. Unlike other immune effector mechanisms that rely on cytolysis, inflammation, release of toxic molecules, or the induction of host cell death, the adaptive immune apparatus prevents Acanthamoeba infections of the cornea by simply preventing the attachment of the parasite to the epithelial surface. The beauty of this mechanism lies in its exquisite simplicity and efficacy.

  15. The Effects of Surgical Factors on Postoperative Astigmatism in Patients Enrolled in the Infant Aphakia Treatment Study (IATS)

    PubMed Central

    Wall, Palak B.; Lee, Jason A.; Lynn, Michael; Lambert, Scott R.; Traboulsi, Elias I.

    2015-01-01

    Purpose To evaluate the impact of surgical factors such as incision type, number of sutures, and technique of closure on postoperative astigmatism in infants undergoing cataract extraction with or without intraocular lens implantation. Methods The IATS is a multicenter (n=12) clinical trial in which 114 infants with unilateral congenital cataracts were randomized to undergo cataract extraction with intraocular lens (IOL) placement or contact lens aphakic correction. Surgical videos were reviewed with regard to incision type and location, whether the incision was extended, the number of sutures placed, and technique of closure. Corneal astigmatism was measured using a handheld keratometer prior to surgery and at 1 year of age. Results Corneal astigmatism decreased from a mean of 1.92 D at baseline to 1.62 D at age 1 year in the CL group, but remained almost unchanged from 2.00 D to 2.09 D in the IOL group (p=0.023). There was no statistical difference between the amount of corneal astigmatism irrespective of incision type (p=0.214) and no increase in astigmatism with extension of the incision to facilitate IOL placement (p=0.849) at 1 year. The number of sutures and technique of closure did not influence the amount of astigmatism at 1 year. Conclusions At the age of one year following cataract extraction in infants, contact lens correction and the lack of IOL placement are associated with a significant decrease in postoperative corneal astigmatism compared to intraocular lens placement. No other surgical factors considered in this study had a statistically significant impact on corneal astigmatism. PMID:25266831

  16. Corneal topographic changes and surgically induced astigmatism following combined phacoemulsification and 25-gauge vitrectomy

    PubMed Central

    Sayed, Khulood Mohammed; Farouk, Mahmoud M.; Katome, Takashi; Nagasawa, Toshihiko; Naito, Takeshi; Mitamura, Yoshinori

    2017-01-01

    AIM To evaluate corneal topographic changes and surgically induced astigmatism (SIA) after combined phacoemulsification and 25-gauge transconjunctival sutureless vitrectomy (25-G TSV). METHODS A retrospective study on 96 eyes of 87 patients who underwent combined phacoemulsification and 25-G TSV. The different topographic parameters and SIA were analyzed pre- and postoperatively. RESULTS There was no significant changes in corneal topographic parameters at different follow up periods. Only surface regularity index changed significantly in the 2nd postoperative week and then returned to baseline values thereafter. Mean SIA gradually decreased to reach 0.12 D by the 6th postoperative month. CONCLUSION Corneal surface and astigmatic changes are insignificant in either early or late postoperative periods following combined phacoemulsification and 25-G TSV. The SIA was the minimum among previous reports on sutureless vitrectomy alone or combined with phacoemulsification. Improvement of SIA did not stop at the 3rd postoperative month but it continued till the 6th month postoperatively. PMID:28149780

  17. Autorefraction, Retinoscopy, Javal's Rule, and Grosvenor's Modified Javal's Rule: The Best Predictor of Refractive Astigmatism

    PubMed Central

    Asiedu, Kofi; Ampiah, Emmanuel Ekow

    2016-01-01

    The aim of the study was to determine the level of agreement between Javal's rule, autorefraction, retinoscopy, and refractive astigmatism and to determine which technique is the most suitable substitute when subjective refraction is not applicable using a clinical sample. A total of 36 subjects, 14 males and 22 females, were involved in this study. The intraclass correlation coefficients between subjective refraction, autorefraction, and retinoscopy were 0.895 and 0.989, respectively, for the spherical equivalent. The Bland-Altman 95% limits of agreement between subjective refraction and autorefraction; subjective refraction and retinoscopy; and autorefraction and retinoscopy were −2.84 to 3.58, −0.88 to 1.12, and −3.01 to 3.53, respectively, for the spherical equivalent. The intraclass correlation coefficients between spectacle total astigmatism and the following techniques were as follows: retinoscopy (0.85); autorefraction (0.92); Javal's rule (0.82); and Grosvenor et al. version (0.85). The Bland-Altman 95% limits of agreement between subjective refraction and autorefraction; subjective refraction and retinoscopy; subjective refraction and Javal's rule; and subjective refraction and Grosvenor et al. version were −0.87 to 1.25, −1.49 to 1.99, −0.73 to 1.93, and −0.89 to 1.7, respectively, for the total astigmatism. The study showed that autorefraction and Javal's rule may provide a starting point for subjective refraction cylinder power determination but only retinoscopy may satisfactorily replace subjective refraction total astigmatism when subjective refraction is not applicable. PMID:27803811

  18. Effectiveness of Toric Orthokeratology in the Treatment of Patients with Combined Myopia and Astigmatism

    PubMed Central

    Lyu, Byul; Hwang, Kyu Yeon; Kim, Sun Young; Kim, Su Young

    2016-01-01

    Purpose The purpose of this multi-institute, single-group clinical trial was to evaluate the effectiveness and safety of toric orthokeratology lenses for the treatment of patients with combined myopia and astigmatism. Methods A total of 44 patients were included in this clinical trial. The patients ranged in age from 7 to 49 years, with myopia of -0.75 to -6.0 diopters (D) and astigmatism of 1.25 to 4.0 D. After excluding 21 subjects, 23 subjects (39 eyes) were analyzed after toric orthokeratology lens use. The subjects underwent ophthalmologic examination after 1 day and 1, 2, 3, and 4 weeks of wearing overnight toric orthokeratology lenses. Results A total of 19 subjects (31 eyes) completed the trial after five subjects (eight eyes) dropped out. In the patients who completed the study by wearing lenses for 4 weeks, the myopic refractive error decreased significantly by 2.60 ± 2.21 D (p < 0.001), from -3.65 ± 1.62 to -1.05 ± 1.64 D. The astigmatic refractive error were also significantly decreased by 0.63 ± 0.98 D (p = 0.001), from 2.07 ± 0.83 to 1.44 ± 0.99 D. The mean uncorrected and corrected visual acuities before wearing the lenses were 2.14 ± 0.80 logarithm of the logMAR (logMAR) and 0.05 ± 0.13 logMAR, respectively, which changed to 0.12 ± 0.30 logarithm of the logMAR (p < 0.001) and 0.01 ± 0.04 logMAR (p = 0.156) after 4 weeks. No serious adverse reactions were reported during the clinical trial. Conclusions Our results suggest that toric orthokeratology is an effective and safe treatment for correcting visual acuity in patients with combined myopia and astigmatism. PMID:27980362

  19. Effect of bacteria on survival and growth of Acanthamoeba castellanii.

    PubMed

    Wang, X; Ahearn, D G

    1997-04-01

    The growth and survival of Acanthamoeba castellanii in the presence of Gram-negative bacteria such as Pseudomonas aeruginosa, Escherichia coli, Serratia marcescens, and Stenotrophomonas maltophilia varied with the densities and species of bacteria. All species of bacteria suspended in a buffered saline at densities of 10(5) to 10(6)/ml supported the growth and survival of 10(6)/ml trophozoites of Acanthamoeba castellanii in a buffered saline solution. At densities of bacteria to amoebae of 100:1 or greater, growth and survival of A. castellanii were suppressed, particularly by P. aeruginosa. In an enrichment medium, the rapid growth of most co-inoculated bacteria inhibited the growth and survival of the amoeba.

  20. Three-dimensional solution structure of Acanthamoeba profilin-I

    PubMed Central

    1993-01-01

    We have determined a medium resolution three-dimensional solution structure of Acanthamoeba profilin-I by multidimensional nuclear magnetic resonance spectroscopy. This 13-kD actin binding protein consists of a five stranded antiparallel beta sheet flanked by NH2- and COOH-terminal helices on one face and by a third helix and a two stranded beta sheet on the other face. Data from actin-profilin cross- linking experiments and the localization of conserved residues between profilins in different phyla indicate that actin binding occurs on the molecular face occupied by the terminal helices. The other face of the molecule contains the residues that differ between Acanthamoeba profilins-I and II and may be important in determining the difference in polyphosphoinositide binding between these isoforms. This suggests that lipids and actin bind to different faces of the molecule. PMID:8397216

  1. Polyphenol oxidase produced during encystation of Acanthamoeba castellanii.

    PubMed

    Sykes, D E; Band, R N

    1985-08-01

    Acanthamoeba castellanii has a phenol oxidase activity that is believed to be a laccase. Enzyme activity was found in the outer cyst wall, in the cytoplasm of encysting amoebae and in the encystment medium. Encystment procedures were modified to promote an increase in the amount of soluble enzyme secreted during encystation. Acanthamoeba polyphenol oxidase has a pH optimum of 6.0 and a Km value of 0.21 mM with dihydroxyphenylalanine. The enzyme does not oxidize tyrosine, and it is inhibited by chloride but not by inhibitors of peroxidase. Its synthesis coincides with encystation, and known inhibitors of polyphenol oxidase prevent encystation. Polyphenol oxidase may have a role in making the cyst resistant to mechanical and chemical breakdown.

  2. Detection of glycoproteins in the Acanthamoeba plasma membrane

    SciTech Connect

    Paatero, G.I.L. ); Gahmberg, C.G. )

    1988-11-01

    In the present study the authors have shown that glycoproteins are present in the plasma membrane of Acanthamoeba castellanii by utilizing different radioactive labeling techniques. Plasma membrane proteins in the amoeba were iodinated by {sup 125}I-lactoperoxidase labeling and the solubilized radiolabeled glycoproteins were separated by lectin-Sepharose affinity chromatography followed by polyacrylamide gel electrophoresis. The periodate/NaB{sup 3}H{sub 4} and galactose oxidase/NaB{sup 3}H{sub 4} labeling techniques were used for labeling of surface carbohydrates in the amoeba. Several surface-labeled glycoproteins were observed in addition to a diffusely labeled region with M{sub r} of 55,000-75,000 seen on electrophoresis, which could represent glycolipids. The presence of glycoproteins in the plasma membrane of Acanthamoeba castellanii was confirmed by metabolic labeling with ({sup 35}S)methionine followed by lectin-Sepharose affinity chromatography and polyacrylamide gel electrophoresis.

  3. Molecular and physiological differentiation between pathogenic and nonpathogenic Acanthamoeba.

    PubMed

    Khan, Naveed A; Jarroll, Edward L; Paget, Timothy A

    2002-09-01

    In this study, 14 isolates of Acanthamoeba from both clinical and environmental sources belonging to seven different species were assayed for tolerance of high osmotic pressure, temperature tolerance, extracellular proteases, and cytopathic effects (CPE) on immortalized rabbit corneal epithelial cells. On the basis of the results, amoeba isolates were divided into pathogenic and nonpathogenic groups. Ribosomal DNA sequencing was performed on these isolates. Phylogenetic relationships revealed that all the pathogenic strains tested clustered together as one group, while nonpathogenic strains clustered into other groups. Sequence comparisons with previously published sequences determined that among the six new pathogenic isolates used in this study, five belong to T4 genotype and one to T11. This is the first report of a T11 genotype being found in Acanthamoeba keratitis.

  4. Anti-Acanthamoeba activity of contact lens solutions

    PubMed Central

    Niszl, I.; Markus, M.

    1998-01-01

    AIMS—This study was undertaken to investigate the effects of contact lens disinfecting solutions on strains of Acanthamoeba from the United Kingdom and southern Africa and to compare the results with those of other researchers. No information was previously available for southern African isolates.
METHODS—11 contact lens solutions were tested on cysts of 10 strains of Acanthamoeba.
RESULTS—Not all solutions used in the study were effective, with some for hard and gas permeable contact lenses being more satisfactory than those for soft contact lenses. The most effective of the gas permeable and hard contact lens solutions tested was Transoak (0.01% (wt/vol) benzalkonium chloride), which killed cysts of all strains within 4 hours of exposure. Oxysept 1 (31 mg hydrogen peroxide/ml) was the best soft contact lens solution tested. It eliminated cysts of certain strains within 4 hours, whereas cysts of other strains were only inactivated within either 8 or 72 hours.
CONCLUSIONS—Manufacturers should be aware of the killing time for Acanthamoeba by contact lens solutions and should provide appropriate guidelines for the use thereof. The killing time for cysts of the African and UK isolates studied is, in general, similar. Therefore, it must in the present state of knowledge be assumed that usage guidelines suggested in the UK are also appropriate for travellers to South Africa and for local residents in South Africa.

 Keywords: contact lenses; Acanthamoeba; keratitis PMID:9893594

  5. Chlorhexidine Monotherapy with Adjunctive Topical Corticosteroids for Acanthamoeba Keratitis

    PubMed Central

    Rahimi, Firoozeh; Hashemian, Seyed Mohammad Nasser; Tafti, Mohammadreza Falah; Mehjerdi, Mohammadali Zare; Safizadeh, Mona Seyed; Pour, Elias Khalili; Sefidan, Bahram Bohrani

    2015-01-01

    Purpose: To assess the efficacy of chlorhexidine monotherapy for Acanthamoeba keratitis, and to determine the therapeutic outcomes of concomitant topical corticosteroids. Methods: In this prospective interventional case series, 31 eyes of 31 patients with Acanthamoeba keratitis (AK) were treated with chlorhexidine 0.02% as monotherapy, from April 2010 to April 2011. The diagnosis of AK was made based on clinical manifestations and positive confocal microscopic (confoscan 3.4, Nidek Co. Ltd., Gamagori, Japan) results. We report the percentage of a favorable clinical response within two weeks of initiating treatment, worsening of the infection while receiving chlorhexidine, recovery of visual acuity (VA), duration of treatment with chlorhexidine and corticosteroids, necessity for addition of other anti-Acanthamoeba agents, presence of corneal scar at the end of the treatment, and need for penetrating keratoplasty (PK). Results: Two weeks after initiation of chlorhexidine, improvement in signs and symptoms was observed in 26 (83.9%) patients but 3 eyes required the addition of propamidine. After initial improvement in one patient, the infection worsened, necessitating the addition of Polyhexamethylene Biguanide (PHMB) and propamidine. A total of 26 (83.9%) patients received topical corticosteroids with mean duration of 65.8 ± 45.1 days. In 22 (71%) eyes, final visual acuity was ≥0.80. Improved VA occurred in 29 eyes (93.5%). Optical PK was considered in 3 (9.7%) eyes and a corneal scar developed in 8 (25.8%) eyes. Conclusion: Chlorhexidine is effective for monotherapy in AK and could be a good choice for initiating treatment. After the initial response to anti-Acanthamoeba agents, corticosteroids can be used as adjunctive therapy depending on the clinical condition. PMID:26425310

  6. Activities of Therapeutic Agents and Myristamidopropyl Dimethylamine against Acanthamoeba Isolates

    PubMed Central

    Kilvington, Simon; Hughes, Reanne; Byas, James; Dart, John

    2002-01-01

    The activities of therapeutic agents and myristamidopropyl dimethylamine (MAPD) against Acanthamoeba strains recalcitrant to medical therapy were studied. MAPD minimum cysticidal concentrations were 6.25 to 25 μg/ml; 10 to 30 μg/ml gave at least a 3-log cyst kill after 6 h, and 50 and 100 μg/ml gave at least a 3-log cyst kill within 2 and 1 h, respectively. PMID:12019127

  7. Activities of therapeutic agents and myristamidopropyl dimethylamine against Acanthamoeba isolates.

    PubMed

    Kilvington, Simon; Hughes, Reanne; Byas, James; Dart, John

    2002-06-01

    The activities of therapeutic agents and myristamidopropyl dimethylamine (MAPD) against Acanthamoeba strains recalcitrant to medical therapy were studied. MAPD minimum cysticidal concentrations were 6.25 to 25 microg/ml; 10 to 30 microg/ml gave at least a 3-log cyst kill after 6 h, and 50 and 100 microg/ml gave at least a 3-log cyst kill within 2 and 1 h, respectively.

  8. Use of multiple immunosuppressive agents in recalcitrant ACANTHAMOEBA scleritis.

    PubMed

    Igras, Estera; Murphy, Conor

    2015-04-15

    A 48-year-old woman who is a contact lens wearer presented with unilateral ACANTHAMOEBA keratitis, confirmed by PCR, which responded initially to topical polyhexamethylene biguanide (PHMB) and brolene. Three months later, despite continued treatment, she developed diffuse anterior scleritis with severe pain and marked scleral injection but without evidence of recurrence keratitis. Oral non-steroidal anti-inflammatories and oral high-dose corticosteroids were added without success. Subsequent treatment with intravenous methylprednisolone and high-dose cyclosporine led to a temporary improvement. Re-presenting with signs of recurrent scleritis and severe pain, the antitumor necrosis factor monoclonal antibody adalimumab, and later oral cyclophosphamide, were added. This led to complete quiescence of the scleritis. Unfortunately, frequent recurrences of ACANTHAMOEBA keratitis and anterior uveitis occurred on immunosuppression requiring continued treatment with PHMB, brolene and topical corticosteroids. This is the first case of severe refractory ACANTHAMOEBA scleritis requiring the concomitant use of four immunosuppressive agents to achieve continued disease control. The challenges in managing this case are discussed.

  9. Genotypic characterization of amoeba isolated from Acanthamoeba keratitis in Poland.

    PubMed

    Derda, Monika; Solarczyk, Piotr; Cholewiński, Marcin; Hadaś, Edward

    2015-03-01

    Free-living amoebae belonging to the genus Acanthamoeba are the causative factor of many diseases. Among others, they cause Acanthamoeba keratitis (AK), a condition that usually occurs in contact lens wearers, though it is also observed in non-wearers. The number of diagnosed cases of AK increased more than eightfold during 8 years in the USA, and a proportional increase in frequency also occurred in Poland and Europe. Cases of AK are usually diagnosed late, and their therapy is difficult and rarely successful. AK is an uncommon diagnosis in Poland. The increased number of positive cases observed in our laboratory may reflect the growing at-risk population of contact lens wearers. Acanthamoeba as a genus of facultative human parasites is currently classified into 17 genotypes. Isolates belonging to seven genotypes were found to be associated with AK. One genotype in particular, T4, was found to be overrepresented in human disease. The main finding of our study is that in Poland, AK is almost always associated with the T4 genotype.

  10. Rapid detection and simultaneous molecular profile characterization of Acanthamoeba infections.

    PubMed

    Goldschmidt, Pablo; Degorge, Sandrine; Benallaoua, Djida; Batellier, Laurence; Di Cave, David; Chaumeil, Christine

    2012-10-01

    Diagnosis of Acanthamoeba by microscopic examination, culture, and polymerase chain reactions (PCRs) has several limitations (sensitivity, specificity, lack of detection of several strains, cost of testing for discrimination among strains). We developed a new high-resolution melting real-time PCR (HRM) to detect and characterize Acanthamoeba infections. HRM performances were evaluated with strains from the American Type Culture Collection (ATCC) and with 20 corneal scrapings. The DNA extracted from specimens were amplified, detected, and characterized in 1 run using 2 original primers diluted in a solution containing an intercalating dye. Detection and molecular characterization of Acanthamoeba infections could be achieved in less than 2.5 h with a dramatic reduction in cost of reactants (postamplification procedures and radioactive or fluorescent-labeled molecular probes were unnecessary). HRM detection limits were 0.1 cyst/μL or less (including genotypes T5 and T11), and its sensitivity and specificity were higher than other molecular tests. For the tested strains from the ATCC, the HRM drafted 4 different profiles: Type I (genotypes T2 and T4), Type II (T5 and T7), Type III (T8), and Type IV (T1, T3, T6, T9, T11, T12, and T13).

  11. Partial characterization of Acanthamoeba castellanii (T4 genotype) DNase activity.

    PubMed

    Iqbal, Junaid; Panjwani, Shamvil; Siddiqui, Ruqaiyyah; Khan, Naveed Ahmed

    2015-02-01

    The deoxyribonuclease (DNase) activities of Acanthamoeba castellanii belonging to the T4 genotype were investigated. Using zymographic assays, the DNase activities had approximate molecular masses of 25 and 35 kDa. A. castellanii DNases exhibited activity at wide-ranging temperature of up to 60 °C and at pH ranging from 4 to 9. The DNases activities were unaffected by proteinase-K treatment, divalent cations such as Ca(++), Cu(++), Mg(++), and Zn(++), or divalent cation chelating agent ethylenediaminetetraacetic acid (EDTA) or sodium dodecyl sulfate (SDS). The non-reliance on divalent cations and homology data suggests that A. castellanii DNases belong to the class of eukaryotic lysosomal DNase II but exhibit robust properties. The DNases activity in A. castellanii interfered with the genomic DNA extraction. Extraction methods involving EDTA, SDS, and proteinase-K resulted in low yield of genomic DNA. On the other hand, these methods resulted in high yield of genomic DNA from human cells suggesting the robust nature of A. castellanii DNases that are unaffected by reagents normally used in blocking eukaryotic DNases. In contrast, the use of chaotropic agent such as guanidine thiocyanate improved the yield of genomic DNA from A. castellanii cells significantly. Further purification and characterization of Acanthamoeba DNases is needed to study their non-classic distinct properties and to determine their role in the biology, cellular differentiation, cell cycle progression, and arrest of Acanthamoeba.

  12. Characterization and pathogenetic role of proteinase from Acanthamoeba castellanii.

    PubMed

    Na, B K; Kim, J C; Song, C Y

    2001-01-01

    A secreted proteinase was purified from the culture supernatant of Acanthamoeba castellanii with several chromatographic steps. The purified proteinase was a chymotrypsin-like serine proteinase. Its molecular weight was approximately 12 kDa on SDS-PAGE, and its native molecular weight was 12 kDa when determined by molecular sieve chromatography. It showed a broad temperature optimum ranging 30-55 degrees C with an optimal at 55 degrees C and an optimal pH of 8.5. It could degrade various protein substrates, such as collagen, fibronectin, laminin, secretory immunoglobulin A, immunoglobulin G, plasminogen, fibrinogen, haemoglobin and rabbit corneal proteins. It showed strong cytopathic effects in cultured cells, including HEp2 and HEK cells. The corneal lesions, induced by both the purified proteinase and A. castellanii, displayed similar clinical results for both cases, in which the stromal infiltration and opacity with the epithelial defect were revealed. These results suggest that the enzyme was highly associated with the pathogenesis of Acanthamoeba. The fact that cytopathic effects and development of corneal lesions caused by the proteinase of Acanthamoeba were inhibited by the proteinase inhibitor suggest that the proteinase inhibitor might be useful as a therapeutic agent.

  13. Uptake and Replication of Salmonella enterica in Acanthamoeba rhysodes

    PubMed Central

    Tezcan-Merdol, Dilek; Ljungström, Marianne; Winiecka-Krusnell, Jadwiga; Linder, Ewert; Engstrand, Lars; Rhen, Mikael

    2004-01-01

    The ability of salmonellae to become internalized and to survive and replicate in amoebae was evaluated by using three separate serovars of Salmonella enterica and five different isolates of axenic Acanthamoeba spp. In gentamicin protection assays, Salmonella enterica serovar Dublin was internalized more efficiently than Salmonella enterica serovar Enteritidis or Salmonella enterica serovar Typhimurium in all of the amoeba isolates tested. The bacteria appeared to be most efficiently internalized by Acanthamoeba rhysodes. Variations in bacterial growth conditions affected internalization efficiency, but this effect was not altered by inactivation of hilA, a key regulator in the expression of the invasion-associated Salmonella pathogenicity island 1. Microscopy of infected A. rhysodes revealed that S. enterica resided within vacuoles. Prolonged incubation resulted in a loss of intracellular bacteria associated with morphological changes and loss of amoebae. In part, these alterations were associated with hilA and the Salmonella virulence plasmid. The data show that Acanthamoeba spp. can differentiate between different serovars of salmonellae and that internalization is associated with cytotoxic effects mediated by defined Salmonella virulence loci. PMID:15184177

  14. Assessment of aberrations and visual quality differences between myopic and astigmatic eyes before and after contact lens application

    PubMed Central

    Demir, Mustafa; Kurna, Sevda Aydin; Sengor, Tomris; Atakan, Tugba Gencaga; Sahin, Tayfun

    2015-01-01

    OBJECTIVE: To evaluate the aberration and visual quality differences between myopic and astigmatic eyes before and after contact lens application by using corneal aberrometer and low- contrast sensitivity chart. METHODS: Eighty eyes of 40 patients were included in this study. Patients were divided into two groups as myopic (40 eyes, n=20) and astigmatic groups (40 eyes, n=20). We used aspheric Balafilcon A (Purevision and Purevision Toric Bausch&Lomb, Rochester, USA) lenses for each group. Corneal aberrations and low-contrast sensitivity values were measured and compared for each patient in both groups. RESULTS: There were no statistically significant differences between myopic and astigmatic groups when we compared low-contrast sensitivity values for both on- and off-eyes. Mean total higher-order aberration (HOA) values for off-eye, were 0.29±0.10 μm, and 0.33±0.10 μm for on-eye in the myopic group, while they were 0.42±0.14 μm in off-eye and 0.37±0.23 μm in on-eye in the astigmatic group. Off-eye mean coma, irregular astigmatism and total higher-order aberration RMS (root-mean-square) values were significantly higher in the astigmatic group compared to the myopic group (p=0.006, p=0.001, p=0.001) but mean on-eye RMS values were not. CONCLUSION: Myopic and astigmatic patients differ in terms of high-order aberrations and these differences cannot be equalized after contact lens application, but visual quality can be improved in both patients by using contact lenses. PMID:28058332

  15. Development of an immunochromatographic assay kit using fluorescent silica nanoparticles for rapid diagnosis of Acanthamoeba keratitis.

    PubMed

    Toriyama, Koji; Suzuki, Takashi; Inoue, Tomoyuki; Eguchi, Hiroshi; Hoshi, Saichi; Inoue, Yoshitsugu; Aizawa, Hideki; Miyoshi, Kazutomi; Ohkubo, Michio; Hiwatashi, Eiji; Tachibana, Hiroshi; Ohashi, Yuichi

    2015-01-01

    We developed an immunochromatographic assay kit that uses fluorescent silica nanoparticles bound to anti-Acanthamoeba antibodies (fluorescent immunochromatographic assay [FICGA]) and evaluated its efficacy for the detection of Acanthamoeba and diagnosis of Acanthamoeba keratitis (AK). The sensitivity of the FICGA kit was evaluated using samples of Acanthamoeba trophozoites and cysts diluted to various concentrations. A conventional immunochromatographic assay kit with latex labels (LICGA) was also evaluated to determine its sensitivity in detecting Acanthamoeba trophozoites. To check for cross-reactivity, the FICGA was performed by using samples of other common causative pathogens of infectious keratitis, such as Pseudomonas aeruginosa, Staphylococcus aureus, Staphylococcus epidermidis, and Candida albicans. Corneal scrapings from patients with suspected AK were tested with the FICGA kit to detect the presence of Acanthamoeba, and the results were compared with those of real-time PCR. The FICGA kit detected organisms at concentrations as low as 5 trophozoites or 40 cysts per sample. There were no cross-reactivities with other pathogens. The FICGA was approximately 20 times more sensitive than the LICGA for the detection of Acanthamoeba trophozoites. The FICGA kit yielded positive results for all 10 patients, which corresponded well with the real-time PCR results. The FICGA kit demonstrated high sensitivity for the detection of Acanthamoeba and may be useful for the diagnosis of AK.

  16. Isolation of Acanthamoeba from the rhizosphere of maize and lucerne plants.

    PubMed

    Orosz, Erika; Farkas, Agnes; Ködöböcz, László; Becságh, Péter; Danka, József; Kucsera, István; Füleky, György

    2013-03-01

    Acanthamoeba species are free-living amoebae that can be found in almost every range of environments. Within this genus, a number of species are recognized as human pathogens, potentially causing Acanthamoeba keratitis, granulomatous amoebic encephalitis, and chronic granulomatous lesions. Soil and water samples were taken from experimental station at Julianna Major of Plant Protection Institute of Centre for Agricultural Research, Hungarian Academy of Sciences (CAR HAS). We detected living Acanthamoeba spp. based on culture-confirmed detection combined with the molecular taxonomic identification method. Living Acanthamoeba spp. were detected in thirteen (65%) samples. The presence of Acanthamoeba spp. in the samples depends significantly on the rhizosphere plants. The most frequently identified living Acanthamoeba genotype was T4 followed by T11, T2/T6 and T17. Genotypes T4 and T11 of Acanthamoeba, are responsible for Acanthamoeba keratitis as well as granulomatous amoebic encephalitis, and should therefore be considered as a potential health risk associated with human activities in the environment.

  17. Microarray analysis of differentially expressed genes between cysts and trophozoites of Acanthamoeba castellanii.

    PubMed

    Moon, Eun-Kyung; Xuan, Ying-Hua; Chung, Dong-Il; Hong, Yeonchul; Kong, Hyun-Hee

    2011-12-01

    Acanthamoeba infection is difficult to treat because of the resistance property of Acanthamoeba cyst against the host immune system, diverse antibiotics, and therapeutic agents. To identify encystation mediating factors of Acanthamoeba, we compared the transcription profile between cysts and trophozoites using microarray analysis. The DNA chip was composed of 12,544 genes based on expressed sequence tag (EST) from an Acanthamoeba ESTs database (DB) constructed in our laboratory, genetic information of Acanthamoeba from TBest DB, and all of Acanthamoeba related genes registered in the NCBI. Microarray analysis indicated that 701 genes showed higher expression than 2 folds in cysts than in trophozoites, and 859 genes were less expressed in cysts than in trophozoites. The results of real-time PCR analysis of randomly selected 9 genes of which expression was increased during cyst formation were coincided well with the microarray results. Eukaryotic orthologous groups (KOG) analysis showed an increment in T article (signal transduction mechanisms) and O article (posttranslational modification, protein turnover, and chaperones) whereas significant decrement of C article (energy production and conversion) during cyst formation. Especially, cystein proteinases showed high expression changes (282 folds) with significant increases in real-time PCR, suggesting a pivotal role of this proteinase in the cyst formation of Acanthamoeba. The present study provides important clues for the identification and characterization of encystation mediating factors of Acanthamoeba.

  18. Characterisation and expression analysis of trophozoite and cyst proteins of Acanthamoeba spp. isolated from Acanthamoeba keratitis (AK) patient.

    PubMed

    Behera, Himansu Sekhar; Satpathy, Gita

    2016-01-01

    The study was carried out to characterise and analyze the expression pattern of proteins of infective trophozoite and cyst forms of Acanthamoeba spp. isolated from an amoebic keratitis patient. Protein was isolated from the trophozoites and cysts of Acanthamoeba spp. isolates and subjected to SDS PAGE, 2D PAGE analysis where a large number of protein bands and protein spots were observed. Four prominent protein spots i.e. 2 from trophozoites and 2 from cysts that appeared more intense compared to the corresponding spots in other corresponding gel were excised from the 2D PAGE gels and analysed by MALDI-TOF/TOF MS assay and Mascot search software. Protein spots from trophozoites were identified as "hypothetical protein ACA1" and "eukaryotic porin protein" and those from cysts were identified as "chaperone protein DnaK" and "chaperonin protein" respectively. Proteomic results of 4 proteins were further validated by reverse genomics using quantitative real time PCR assay which showed a 1388 fold and 4.35 fold increase in expression of "hypothetical protein ACA1" gene and "eukaryotic porin protein" gene respectively in trophozoites compared to cysts and a 15 fold and 12.36 fold increase in expression of "chaperone protein DnaK" gene and "chaperonin protein" gene respectively in cysts compared to trophozoites. "Hypothetical protein ACA1" of trophozoites, whose function is unknown might have some important role in the parasite division and pathogenicty of Acanthamoeba spp. which needs further study. As trophozoites are the active and feeding form of Acanthamoeba spp., "eukaryotic porin" proteins may have some important role in efflux of toxic metabolites and exudates from interior of cell to outside along with some role in pathogenicity. Similarly proteins such as "chaperone protein DnaK" and "chaperonin protein" which belongs to group of heat shock proteins may have a role in folding of cyst specific proteins in cyst which needs further study.

  19. Voriconazole as a first-line treatment against potentially pathogenic Acanthamoeba strains from Peru.

    PubMed

    Cabello-Vílchez, Alfonso Martín; Martín-Navarro, Carmen M; López-Arencibia, Atteneri; Reyes-Batlle, María; Sifaoui, Ines; Valladares, Basilio; Piñero, José E; Lorenzo-Morales, Jacob

    2014-02-01

    Pathogenic strains of Acanthamoeba genus are the causative agents of fatal granulomatous amoebic encephalitis and a serious sight-threatening infection of the eye known as Acanthamoeba keratitis. In a previous study, Acanthamoeba strains were isolated from nasal swabs collected from healthy individuals in Peru. In the present study, the pathogenic potential of the isolated strains was established based on temperature and osmotolerance assays as well as the secretion rate of extracellular proteases. Based on these experiments, four strains that showed the highest pathogenic potential were selected for sensitivity assays against two molecules (voriconazole and chlorhexidine) which are currently used for the treatment of Acanthamoeba infections. After performing sensitivity and activity assays, it was found that both drugs were active against the tested strains. However, voriconazole showed higher activity against the studied strains compared to chlorhexidine. Therefore, voriconazole should be established as a first-line treatment against Acanthamoeba infections at least in the studied region of Peru.

  20. Isolation and Molecular Characterization of Acanthamoeba Strains from Dental Units in Costa Rica.

    PubMed

    Retana-Moreira, Lissette; Abrahams-Sandí, Elizabeth; Castro-Artavia, Esteban; Fernández-Sánchez, Ana; Castro-Castillo, Alfredo; Reyes-Batlle, María; Lorenzo-Morales, Jacob

    2015-01-01

    Free-living amoebae are protozoa widely distributed in nature, which can be found in a variety of environments. Four genera are recognized as causal agents of infections in humans and animals: Acanthamoeba, Naegleria, Balamuthia, and Sappinia. In this study, the presence of Acanthamoeba in dental units was determined and the isolates obtained were molecularly characterized; osmotolerance and thermotolerance assays were also performed to evaluate multiplication under these conditions, frequently associated with pathogenicity. The morphological analysis and partial sequencing of the 18S rDNA gene revealed the presence of Acanthamoeba genotype T4 in 14% of the units sampled. Osmotolerance and thermotolerance tests were positive for more than 80% of the isolates. Up to date, this is the first study that reports the detection, identification, and genotyping of Acanthamoeba isolated from dental units in Costa Rica and even in Latin-America. Further assays to determine the potential pathogenicity of these Acanthamoeba isolates are underway.

  1. Insights from the DNA databases: approaches to the phylogenetic structure of Acanthamoeba.

    PubMed

    Fuerst, Paul A

    2014-11-01

    Species of Acanthamoeba have been traditionally described using morphology (primarily cyst structure), or cytology of nuclear division (used by Pussard and Pons, 1977). Twenty-plus putative species were proposed based on such criteria. Morphology, however, is often plastic, dependent upon culture conditions. DNA sequences of the nuclear small subunit (18S) rRNA that can be used for the study of the phylogeny of Acanthamoeba have increased from a single sequence in 1986 to more than 1800 in 2013. Some of the patterns of the sequence data for Acanthamoeba are reviewed, and some of the insights that this data illuminates are illustrated. In particular, the data suggest the existence of 20 or more genotypic types, a number not dissimilar to the number of named species of Acanthamoeba. However, molecular studies make clear that the relationship between phylogenetic relatedness and species names as we know them for Acanthamoeba is tenuous at best.

  2. Genotyping of Acanthamoeba isolated from water in recreational areas of Tehran, Iran.

    PubMed

    Nazar, M; Haghighi, A; Niyyati, M; Eftekhar, M; Tahvildar-Biderouni, F; Taghipour, N; Abadi, A; Nazemalhosseini Mojarad, E; Athari, A

    2011-09-01

    A comprehensive survey assessing the presence of Acanthamoeba was conducted on 50 samples from water sources in parks and public squares from 22 municipal districts of Tehran, Iran. The prevalence and genotypes of Acanthamoeba were determined by PCR and the PCR fragments of ribosomal RNA genes sequenced. Sixteen (32%) samples were positive for Acanthamoeba spp. Sequence analysis revealed that the positive isolates belonged to the T4 and T5 genotypes. Fourteen isolates (87.5%) were T4, and two (12.5%) were T5. Acanthamoeba may be a problematic organism for contact lens wearers and for immunocompromised individuals. In Iran, Acanthamoeba keratitis has increased in recent years, mainly due to poor hygiene in contact lens wearers. A thorough survey for the prevalence of this amoeba could have a significant role in prevention of disease. This is the first report of the T5 genotype from water in recreational areas of Tehran.

  3. Rapid and sensitive diagnosis of Acanthamoeba keratitis by loop-mediated isothermal amplification.

    PubMed

    Ge, Z; Qing, Y; Zicheng, S; Shiying, S

    2013-11-01

    A loop-mediated isothermal amplification (LAMP) assay was developed for the detection of Acanthamoeba. The sensitivity of the LAMP assay was tested using different copies of positive DNA. The specificity of the assay was tested using DNA extracted from Acanthamoeba, Pseudomonas aeruginosa, Candida albicans, herpes simplex virus-1 and human corneal epithelial cells. Its effectiveness was evaluated and compared with culture, corneal smear examination and real-time PCR in corneal samples from mice with Acanthamoeba keratitis. We also tested three corneal samples from patients with suspected Acanthamoeba or fungal infection using LAMP. Loop-mediated isothermal amplification was confirmed to be very sensitive, with the lowest detection limit being ten copies/tube of Acanthamoeba DNA. The LAMP primers only amplified Acanthamoeba DNA. During the development of Acanthamoeba keratitis in mice, almost all of the positive rates of LAMP at each time post-infection were higher than those of culture or corneal smear examination. The total positive rate of LAMP was significantly higher than those of culture and corneal smear examination (p <0.05), whereas the sensitivities of LAMP and real-time PCR were comparable. However, the trends of positive change in these different test methods were generally similar. Of the three clinical corneal specimens, two with suspected Acanthamoeba keratitis tested positive for Acanthamoeba using LAMP along with culture or corneal smear examination, whereas the other suspected fungal keratitis tested negative. The LAMP assay is a simple, rapid, highly specific and sensitive method for the diagnosis of keratitis caused by Acanthamoeba.

  4. A unique astigmatic nodal property in misaligned Ritchey-Chrétien telescopes with misalignment coma removed.

    PubMed

    Schmid, Tobias; Thompson, Kevin P; Rolland, Jannick P

    2010-03-01

    We present the aberration field response of Ritchey-Chrétien telescopes, with the aperture stop on the primary mirror, to secondary mirror misalignments. More specifically, we derive a general condition for the geometry of the binodal astigmatic aberration field for a telescope that has been aligned to remove field-constant coma. It has been observed that when the coma caused by secondary mirror misalignments is removed the astigmatic field is typically not symmetric around the periphery, but, significantly, it is always effectively zero on-axis. This observation is a manifestation of binodal astigmatism where one of the astigmatic nodes remains near the field center. Here, we show how the condition to remove field-constant coma simultaneously creates a constraint whereby one of the astigmatic nodes must remain effectively on-axis. This result points to why the alignment of a large telescope based on axial imagery is insufficient and demonstrates exactly the geometry of the remaining misalignment aberration field, which dominates the performance of the telescope, providing insights into more complete alignment approaches.

  5. Identifying differentially expressed genes in trophozoites and cysts of Acanthamoeba T4 genotype: Implications for developing new treatments for Acanthamoeba keratitis.

    PubMed

    Abedkhojasteh, Hoda; Niyyati, Maryam; Rezaei, Sasan; Mohebali, Mehdi; Farnia, Shohreh; Kazemi-Rad, Elham; Roozafzoon, Reza; Sianati, Hamed; Rezaeian, Mostafa; Heidari, Mansour

    2015-02-01

    Acanthamoeba T4 genotype is the most prevalent genotype associated with amoebic keratitis. Acanthamoeba keratitis therapy is difficult due to transformation of trophozoite to cyst stage, which hinders the treatment of the disease. Although encystation assists the organism to survive against the chemotherapeutic compounds, the precise mechanism of encystation remains poorly understood. The purpose of this work was to identify differentially expressed genes in Acanthamoeba T4 genotype which might be useful for understanding of the encystment process and may thus help develop more efficient treatment. The mRNA profile of trophozoite and cyst of Acanthamoeba T4 genotype isolated from a soft contact lens wearer were analyzed using a cDNA amplified fragment length polymorphism (cDNA-AFLP) technique. Subsequently, a real time reverse transcriptase-PCR was performed to validate the cDNA-AFLP results. Three genes, heat shock protein70 (hsp70), actin-I and elongation factor-1alpha (EF-1α) were differentially expressed during Acanthamoeba differentiation. An in silico result predicted that transformation of trophozoite to cyst could be mediated through their cooperation with the protein partners interaction. Taken together, our experimental and bioinformatics findings suggested potential functions of hsp70, EF-1α and actin-I in differentiation of Acanthamoeba T4 genotype which may be useful in the design of an efficient therapeutic strategy in AK.

  6. Visual outcomes of topography-guided excimer laser surgery for treatment of patients with irregular astigmatism.

    PubMed

    Ghoreishi, Mohammad; Naderi Beni, Afsaneh; Naderi Beni, Zahra

    2014-01-01

    The aim of this study was to evaluate the efficacy, safety, and predictability of topography-guided treatments to enhance refractive status following other corneal surgical procedures. In a prospective case series study, 28 consecutive eyes of 26 patients with irregular astigmatism after radial keratotomy, corneal transplant, small hyperopic and myopic excimer laser optical zones, and corneal scars were operated. Laser-assisted in situ keratomileusis (LASIK) (n = 8) and photorefractive keratectomy (PRK) (n = 20) were performed using the ALLEGRETTO WAVE excimer laser and topography-guided customized ablation treatment software. Preoperative and postoperative uncorrected visual acuity (UCVA), best corrected visual acuity (BCVA), manifest and cycloplegic refraction, and corneal topography with asphericity were analyzed in 12 months follow-up. Uncorrected visual acuity (UCVA) changed from 0.2 ± 0.2 or (20/100 ± 20/100) to 0.51 ± 0.31 or (20/40 ± 20/60) in the LASIK group (P = 0.01) and from 0.34 ± 0.16 or (20/60 ± 20/120) to 0.5 ± 0.23 or (20/40 ± 20/80) in the PRK group (P = 0.01). Refractive cylinder decreased from -3.2 ± 0.84 diopters (D) to -2.06 ± 0.42 D in the LASIK group (P = 0.07) and from -2.25 ± 0.39 D to -1.5 ± 0.23 D in the PRK group (P = 0.008). Best corrected visual acuity did not change significantly in either group. Topography-guided treatment is effective in correcting the irregular astigmatism after refractive surgery. Topography-guided PRK can significantly reduce irregular astigmatism and increase the UCVA and BCVA.

  7. Astigmatic multifocus microscopy enables deep 3D super-resolved imaging

    PubMed Central

    Oudjedi, Laura; Fiche, Jean-Bernard; Abrahamsson, Sara; Mazenq, Laurent; Lecestre, Aurélie; Calmon, Pierre-François; Cerf, Aline; Nöllmann, Marcelo

    2016-01-01

    We have developed a 3D super-resolution microscopy method that enables deep imaging in cells. This technique relies on the effective combination of multifocus microscopy and astigmatic 3D single-molecule localization microscopy. We describe the optical system and the fabrication process of its key element, the multifocus grating. Then, two strategies for localizing emitters with our imaging method are presented and compared with a previously described deep 3D localization algorithm. Finally, we demonstrate the performance of the method by imaging the nuclear envelope of eukaryotic cells reaching a depth of field of ~4µm. PMID:27375935

  8. Long-term outcomes of wedge resection at the limbus for high irregular corneal astigmatism after repaired corneal laceration

    PubMed Central

    Du, Jun; Zheng, Guang-Ying; Wen, Cheng-Lin; Zhang, Xiao-Fang; Zhu, Yu

    2016-01-01

    AIM To evaluate the clinical value of wedge resection at corneal limbus in patients with traumatic corneal scarring and high irregular astigmatism. METHODS Patients with traumatic corneal astigmatism received wedge resection at least 6mo after suture removal from corneal wound. The uncorrected distance visual acuities (UCVA) and best corrected distance visual acuities (BCVA), pre- and post-operation astigmatism, spherical equivalent (SE), safety and complications were evaluated. RESULTS Ten eyes (10 patients) were enrolled in this study. Mean follow-up time after wedge resection was 37.8±15.4mo (range, 20-61mo). The mean UCVA improved from +1.07±0.55 logMAR to +0.43±0.22 logMAR (P=0.000) and the mean BCVA from +0.50±0.30 logMAR to +0.15±0.17 logMAR (P=0.000). The mean astigmatism power measured by retinoscopy was -2.03±2.27 D postoperatively and -2.83±4.52 D preoperatively (P=0.310). The mean SE was -0.74±1.61 D postoperatively and -0.64±1.89 D preoperatively (P=0.601). Two cases developed mild pannus near the sutures. No corneal perforation, infectious keratitis or wound gape occurred. CONCLUSION Corneal-scleral limbal wedge resection with compression suture is a safe, effective treatment for poor patients with high irregular corneal astigmatism after corneal-scleral penetrating injury. Retinoscopy can prove particularly useful for high irregular corneal astigmatism when other measurements are not amenable. PMID:27366685

  9. Artemether Exhibits Amoebicidal Activity against Acanthamoeba castellanii through Inhibition of the Serine Biosynthesis Pathway

    PubMed Central

    Deng, Yihong; Ran, Wei; Man, Suqin; Li, Xueping; Gao, Hongjian; Tang, Wei

    2015-01-01

    Acanthamoeba sp. parasites are the causative agents of Acanthamoeba keratitis, fatal granulomatous amoebic encephalitis, and cutaneous infections. However, there are currently no effective drugs for these organisms. Here, we evaluated the activity of the antimalarial agent artemether against Acanthamoeba castellanii trophozoites and identified potential targets of this agent through a proteomic approach. Artemether exhibited in vitro amoebicidal activity in a time- and dose-dependent manner and induced ultrastructural modification and cell apoptosis. The iTRAQ quantitative proteomic analysis identified 707 proteins that were differentially expressed after artemether treatment. We focused on phosphoglycerate dehydrogenase and phosphoserine aminotransferase in the serine biosynthesis pathway because of their importance to the growth and proliferation of protozoan and cancer cells. The expression of these proteins in Acanthamoeba was validated using quantitative real-time PCR and Western blotting after artemether treatment. The changes in the expression levels of phosphoserine aminotransferase were consistent with those of phosphoglycerate dehydrogenase. Therefore, the downregulation of phosphoserine aminotransferase may be due to the downregulation of phosphoglycerate dehydrogenase. Furthermore, exogenous serine might antagonize the activity of artemether against Acanthamoeba trophozoites. These results indicate that the serine biosynthesis pathway is important to amoeba survival and that targeting these enzymes would improve the treatment of Acanthamoeba infections. Artemether may be used as a phosphoglycerate dehydrogenase inhibitor to control or block Acanthamoeba infections. PMID:26014935

  10. Abietane diterpenoids from Salvia sclarea transformed roots as growth inhibitors of pathogenic Acanthamoeba spp.

    PubMed

    Kuźma, Łukasz; Derda, Monika; Hadaś, Edward; Wysokińska, Halina

    2015-01-01

    Amoebae from the genus Acanthamoeba are known agents leading to various diseases such as granulomatous amoebic encephalitis (GAE), a chronic progressive disease of the central nervous system, amoebic keratitis (AK), chronic eye infection, amoebic pneumitis (AP), chronic lung infection, and skin infections. It is known that various synthetic anti-Acanthamoeba substances are ineffective. Therefore, other substances, e.g., natural plant compounds, are the focus of biological investigations regarding anti-parasite activity. In this work, the ability of four abietane diterpenoids (ferruginol, salvipisone, aethiopinone, and 1-oxo-aethiopinone) to inhibit Acanthamoeba growth is reported. All investigated compounds were active against Acanthamoeba growing in vitro. Among them, ferruginol demonstrated the highest activity against Acanthamoeba. This compound inhibited Acanthamoeba growth by about 72% in a 3-day exposure period (IC50 17.45 μM), while aethiopinone and 1-oxo-aethiopinone demonstrated this activity at the level of 55-56%. Salvipisone reduced the growth of Acanthamoeba in vitro culture by 39%. For this compound, the value of IC50 was 701.94 μM after 72 h of exposure.

  11. Loop-mediated isothermal amplification targeting 18S ribosomal DNA for rapid detection of Acanthamoeba.

    PubMed

    Yang, Hye-Won; Lee, Yu-Ran; Inoue, Noboru; Jha, Bijay Kumar; Danne, Dinzouna-Boutamba Sylvatrie; Kim, Hong-Kyun; Lee, Junhun; Goo, Youn-Kyoung; Kong, Hyun-Hee; Chung, Dong-Il; Hong, Yeonchul

    2013-06-01

    Amoebic keratitis (AK) caused by Acanthamoeba is one of the most serious corneal infections. AK is frequently misdiagnosed initially as viral, bacterial, or fungal keratitis, thus ensuring treatment delays. Accordingly, the early detection of Acanthamoeba would contribute significantly to disease management and selection of an appropriate anti-amoebic therapy. Recently, the loop-mediated isothermal amplification (LAMP) method has been applied to the clinical diagnosis of a range of infectious diseases. Here, we describe a rapid and efficient LAMP-based method targeting Acanthamoeba 18S rDNA gene for the detection of Acanthamoeba using clinical ocular specimens in the diagnosis of AK. Acanthamoeba LAMP assays detected 11 different strains including all AK-associated species. The copy number detection limit for a positive signal was 10 DNA copies of 18S rDNA per reaction. No cross-reactivity with the DNA of fungi or other protozoa was observed. The sensitivity of LAMP assay was higher than those of Nelson primer PCR and JDP primer PCR. In the present study, LAMP assay based on directly heat-treated samples was found to be as efficient at detecting Acanthamoeba as DNA extracted using a commercial kit, whereas PCR was only effective when commercial kit-extracted DNA was used. This study showed that the devised Acanthamoeba LAMP assay could be used to diagnose AK in a simple, sensitive, and specific manner.

  12. Molecular detection and genotyping of Acanthamoeba spp. among stray dogs using conjunctival swab sampling.

    PubMed

    Karakuş, Mehmet; Aykur, Mehmet; Özbel, Yusuf; Töz, Seray; Dağcı, Hande

    2016-12-01

    Acanthamoeba is one of the most common free-living amoebas (FLA) that present in environment. In humans, Acanthamoeba can cause an infection of the eye termed Acanthamoeba keratitis, which mostly occurs in contact lens wearers. In the present study, we aimed to screen the presence of Acanthamoeba DNA in stray dogs using previously collected conjunctival swab samples in a hyper-endemic area for canine leishmaniasis. Totally, 184 dogs were included in the study and 27 of them (14.6%) were found positive for Acanthamoeba according to the 18s rRNA gene sequencing. Two different genotypes (T4 and T5) were identified and T5 was firstly reported in Turkey in the present study. Statistical analysis was performed and no correlation was found between Leishmania and Acanthamoeba positivity (P<0.05). To best of our knowledge, this is the first study conducted to screen Acanthamoeba among stray dogs. Further studies are necessary to reveal the infection status and genotypes among dogs and its possible correlation with leishmaniasis.

  13. Artemether Exhibits Amoebicidal Activity against Acanthamoeba castellanii through Inhibition of the Serine Biosynthesis Pathway.

    PubMed

    Deng, Yihong; Ran, Wei; Man, Suqin; Li, Xueping; Gao, Hongjian; Tang, Wei; Tachibana, Hiroshi; Cheng, Xunjia

    2015-08-01

    Acanthamoeba sp. parasites are the causative agents of Acanthamoeba keratitis, fatal granulomatous amoebic encephalitis, and cutaneous infections. However, there are currently no effective drugs for these organisms. Here, we evaluated the activity of the antimalarial agent artemether against Acanthamoeba castellanii trophozoites and identified potential targets of this agent through a proteomic approach. Artemether exhibited in vitro amoebicidal activity in a time- and dose-dependent manner and induced ultrastructural modification and cell apoptosis. The iTRAQ quantitative proteomic analysis identified 707 proteins that were differentially expressed after artemether treatment. We focused on phosphoglycerate dehydrogenase and phosphoserine aminotransferase in the serine biosynthesis pathway because of their importance to the growth and proliferation of protozoan and cancer cells. The expression of these proteins in Acanthamoeba was validated using quantitative real-time PCR and Western blotting after artemether treatment. The changes in the expression levels of phosphoserine aminotransferase were consistent with those of phosphoglycerate dehydrogenase. Therefore, the downregulation of phosphoserine aminotransferase may be due to the downregulation of phosphoglycerate dehydrogenase. Furthermore, exogenous serine might antagonize the activity of artemether against Acanthamoeba trophozoites. These results indicate that the serine biosynthesis pathway is important to amoeba survival and that targeting these enzymes would improve the treatment of Acanthamoeba infections. Artemether may be used as a phosphoglycerate dehydrogenase inhibitor to control or block Acanthamoeba infections.

  14. Loop-Mediated Isothermal Amplification Targeting 18S Ribosomal DNA for Rapid Detection of Acanthamoeba

    PubMed Central

    Yang, Hye-Won; Lee, Yu-Ran; Inoue, Noboru; Jha, Bijay Kumar; Danne, Dinzouna-Boutamba Sylvatrie; Kim, Hong-Kyun; Lee, Junhun; Goo, Youn-Kyoung; Kong, Hyun-Hee; Chung, Dong-Il

    2013-01-01

    Amoebic keratitis (AK) caused by Acanthamoeba is one of the most serious corneal infections. AK is frequently misdiagnosed initially as viral, bacterial, or fungal keratitis, thus ensuring treatment delays. Accordingly, the early detection of Acanthamoeba would contribute significantly to disease management and selection of an appropriate anti-amoebic therapy. Recently, the loop-mediated isothermal amplification (LAMP) method has been applied to the clinical diagnosis of a range of infectious diseases. Here, we describe a rapid and efficient LAMP-based method targeting Acanthamoeba 18S rDNA gene for the detection of Acanthamoeba using clinical ocular specimens in the diagnosis of AK. Acanthamoeba LAMP assays detected 11 different strains including all AK-associated species. The copy number detection limit for a positive signal was 10 DNA copies of 18S rDNA per reaction. No cross-reactivity with the DNA of fungi or other protozoa was observed. The sensitivity of LAMP assay was higher than those of Nelson primer PCR and JDP primer PCR. In the present study, LAMP assay based on directly heat-treated samples was found to be as efficient at detecting Acanthamoeba as DNA extracted using a commercial kit, whereas PCR was only effective when commercial kit-extracted DNA was used. This study showed that the devised Acanthamoeba LAMP assay could be used to diagnose AK in a simple, sensitive, and specific manner. PMID:23864737

  15. Apparatus for and method of correcting for astigmatism in a light beam reflected off of a light reflecting surface

    DOEpatents

    Sawicki, R.H.; Sweatt, W.

    1985-11-21

    A technique for adjustably correcting for astigmatism in a light beam is disclosed herein. This technique defines a flat, rectangular light reflecting surface having opposite reinforced side edges and which is resiliently bendable, to a limited extent, into different concave and/or convex cylindrical curvatures about a particular axis and provides for adjustably bending the light reflecting surface into one of different curvatures depending upon the astigmatism to be corrected and for fixedly maintaining the curvature selected. In the embodiment disclosed, the light reflecting surface is adjustably bendable into the selected cylindrical curvature by application of a particular bending moment to the reinforced side edges of the light reflecting surface.

  16. Optimization of nonimaging focusing heliostat in dynamic correction of astigmatism for a wide range of incident angles.

    PubMed

    Chong, Kok-Keong

    2010-05-15

    To overcome astigmatism has always been a great challenge in designing a heliostat capable of focusing the sunlight on a small receiver throughout the year. In this Letter, a nonimaging focusing heliostat with a dynamic adjustment of facet mirrors in a group manner has been analyzed for optimizing the astigmatic correction in a wide range of incident angles. This what is to the author's knowledge a new heliostat is not only designed to serve the purpose of concentrating sunlight to several hundreds of suns, but also to significantly reduce the variation of the solar flux distribution with the incident angle.

  17. Refractive lenticule extraction (ReLEx) through a small incision (SMILE) for correction of myopia and myopic astigmatism: current perspectives

    PubMed Central

    Ağca, Alper; Demirok, Ahmet; Yıldırım, Yusuf; Demircan, Ali; Yaşa, Dilek; Yeşilkaya, Ceren; Perente, İrfan; Taşkapılı, Muhittin

    2016-01-01

    Small-incision lenticule extraction (SMILE) is an alternative to laser-assisted in situ keratomileusis (LASIK) and photorefractive keratectomy (PRK) for the correction of myopia and myopic astigmatism. SMILE can be performed for the treatment of myopia ≤−12 D and astigmatism ≤5 D. The technology is currently only available in the VisuMax femtosecond laser platform. It offers several advantages over LASIK and PRK; however, hyperopia treatment, topography-guided treatment, and cyclotorsion control are not available in the current platform. The working principles, potential advantages, and disadvantages are discussed in this review. PMID:27757010

  18. Acanthamoeba spp. in Contact Lenses from Healthy Individuals from Madrid, Spain

    PubMed Central

    Gomes, Thiago dos Santos; Magnet, Angela; Izquierdo, Fernando; Vaccaro, Lucianna; Redondo, Fernando; Bueno, Sara; Sánchez, Maria Luisa; Angulo, Santiago; Fenoy, Soledad; Hurtado, Carolina; del Aguila, Carmen

    2016-01-01

    Purpose Acanthamoeba keratitis (AK) is a painful and potentially blinding corneal infection caused by Acanthamoeba spp. In Madrid, environmental studies have demonstrated a high presence of these free-living amoebae in tap water. Since most of AK cases occur in contact lenses (CL) wearers with inadequate hygiene habits, the presence of Acanthamoeba in discarded CL has been studied and compared with other common etiological agents of keratitis, such as Pseudomonas aeruginosa and Staphylococcus aureus. Methods One hundred and seventy-seven healthy individuals from Madrid contributed their discarded CL and answered a questionnaire on hygiene habits. DNA was extracted from the CL solution and analyzed by real-time PCR for Acanthamoeba, Pseudomonas aeruginosa and Staphylococcus aureus. These CL and their solutions were also cultured on non-nutrient agar to isolate Acanthamoeba. Results Among the 177 samples, Acanthamoeba DNA was detected in 87 (49.2%), P. aeruginosa DNA in 14 (7.9%) and S. aureus DNA in 19 (10.7%). Cultivable amoebae, however, were observed in only one sample (0.6%). This isolate was genotyped as T4. The habits reported by this CL owner included some recognized risk factors for AK, but in this study only the practice of “not cleaning the CL case” presented some statistical significant association with Acanthamoeba DNA presence. Detection of the investigated bacterial DNA did not demonstrate statistical significant association with the studied practices, but the presence of P. aeruginosa revealed a possible inhibition of Acanthamoeba in these samples. Conclusions The PCR results suggest a high presence of Acanthamoeba spp. in healthy CL wearers from Madrid, but we can assume that CL solutions are properly disinfecting the CL since only 1.1% of the positive PCR samples correspond to viable amoebae and, after four years, only one participant reported stronger ocular problems. Nevertheless, more studies are necessary to corroborate this hypothesis. PMID

  19. Acanthamoeba T3, T4 and T5 in swimming-pool waters from Southern Brazil.

    PubMed

    Caumo, Karin; Rott, Marilise B

    2011-03-01

    Species of Acanthamoeba, known to cause keratitis (AK) and granulomatous encephalitis in humans are frequently isolated from a variety of water sources. In this study, 13 Acanthamoeba isolates from swimming pools were classified at the genotype level based on the sequence analysis of the Acanthamoeba small-subunit rRNA gene. Nine of the 13 isolates were genotype T5, three were genotype T4, and one was T3. Several genotypes have been reported worldwide as causative agents of AK, including genotypes T3, T4, and T5. The present study indicates that genotype T5 is a common contaminant in swimming-pool water.

  20. Pathogenic and nonpathogenic Acanthamoeba spp. in thermally polluted discharges and surface waters

    SciTech Connect

    de Jonckheere, J.F.

    1981-02-01

    During spring and autumn, the total number of amoebae and the number of acanthamoeba species able to grow at 37 degrees C were determined in six thermally polluted factory discharges and the surrounding surface waters. The isolated Acanthamoeba strains were studied for growth in axenic medium, cytopathic effect in Vito cell cultures, and virulence in mice. Although more amoebae were isolated in autumn, the number of Acanthamoeba species was lower than in spring, when the percent of pathogenic strains among the isolates was highest. Higher concentrations of amoebae were found in warm discharges, and more virulent strains occurred in thermal discharges than in surface waters.

  1. Disseminated Acanthamoeba sinusitis in a patient with AIDS: a possible role for early antiretroviral therapy.

    PubMed

    Carter, Wendy W; Gompf, Sandra G; Toney, John F; Greene, John N; Cutolo, Edward P

    2004-01-01

    Acanthamoeba, a free-living ameba, has been reported to infect humans with subacute encephalitis, sinusitis, or keratitis. Multiple cases of Acanthamoeba sinusitis with dissemination have been reported in association with AIDS, with high mortality. We report successful treatment of a 35-year-old woman who presented with sinusitis that progressed to disseminated acanthamebiasis as her initial manifestation of AIDS. To our knowledge, our patient was one of the few and longest-lived survivors of disseminated Acanthamoeba infection with AIDS. As with other opportunistic infections, early aggressive therapy including HAART may alter the outcome in this almost uniformly fatal disease.

  2. Mode-coupling enhancement by pump astigmatism correction in a Ti:Sapphire femtosecond laser.

    PubMed

    Ramírez-Guerra, Catalina; Moreno-Larios, José Agustín; Rosete-Aguilar, Martha; Garduño-Mejía, Jesús

    2016-12-01

    To pump a solid-state femtosecond laser cavity, a beam from a CW laser is focused by a single lens into the laser crystal. To increase the output power of the laser, the overlap of the laser mode with the pump mode should be maximized. This is particularly important in the so-called mode coupling and the Kerr-lens mode locking (KLM) operation, where the change in beam waist at the position of the gain medium is exploited to enhance the mode overlap with the pump laser in the crystal. In this paper, the astigmatism in the pump beam is reduced by tilting the pump lens. A Gaussian beam is propagated through the complete focusing system-pump lens, tilted spherical mirror, and crystal cut at Brewster's angle-to show the astigmatism inside the crystal as a function of the tilt of the pump lens. A genetic algorithm is presented to optimize the mode coupling between the pump and laser beam inside the crystal by tilting the pump lens. Experimental results are presented to verify the design, showing an increase in the output power of the laser cavity of about 20%.

  3. Independent control of beam astigmatism and ellipticity using a SLM for fs-laser waveguide writing.

    PubMed

    Ruiz de la Cruz, A; Ferrer, A; Gawelda, W; Puerto, D; Sosa, M Galván; Siegel, J; Solis, J

    2009-11-09

    We have used a low repetition rate (1 kHz), femtosecond laser amplifier in combination with a spatial light modulator (SLM) to write optical waveguides with controllable cross-section inside a phosphate glass sample. The SLM is used to induce a controllable amount of astigmatism in the beam wavefront while the beam ellipticity is controlled through the propagation distance from the SLM to the focusing optics of the writing set-up. The beam astigmatism leads to the formation of two separate disk-shaped foci lying in orthogonal planes. Additionally, the ellipticity has the effect of enabling control over the relative peak irradiances of the two foci, making it possible to bring the peak irradiance of one of them below the material transformation threshold. This allows producing a single waveguide with controllable cross-section. Numerical simulations of the irradiance distribution at the focal region under different beam shaping conditions are compared to in situ obtained experimental plasma emission images and structures produced inside the glass, leading to a very satisfactory agreement. Finally, guiding structures with controllable cross-section are successfully produced in the phosphate glass using this approach.

  4. Characterization of a tunable astigmatic fluidic lens with adaptive optics correction for compact phoropter application

    NASA Astrophysics Data System (ADS)

    Fuh, Yiin-Kuen; Huang, Chieh-Tse

    2014-07-01

    Fluidically controlled lenses which adaptively correct prescribed refractive error without mechanically moving parts are extensively applied in the ophthalmic applications. Capable of variable-focusing properties, however, the associated aberrations due to curvature change and refractive index mismatch can inherently degrade image quality severely. Here we present the experimental study of the aberrations in tunable astigmatic lens and use of adaptive optics to compensate for the wavefront errors. Characterization of the optical properties of the individual lenses is carried out by Shack-Hartmann measurements. An adaptive optics (AO) based scheme is demonstrated for three injected fluidic volumes, resulting in a substantial reduction of the wavefront errors from -0.12, -0.25, -0.32 to 0.01, -0.01, -0.20 μm, respectively, corresponding to the optical power tenability of 0.83 to 1.84 D. Furthermore, an integrated optical phoroptor consisting of adjustable astigmatic lenses and AO correction is demonstrated such that an induced refraction error of -1 D cylinder at 180° of a model eye vision is experimentally corrected.

  5. Proteomic profiling of the infective trophozoite stage of Acanthamoeba polyphaga.

    PubMed

    Caumo, Karin Silva; Monteiro, Karina Mariante; Ott, Thiely Rodrigues; Maschio, Vinicius José; Wagner, Glauber; Ferreira, Henrique Bunselmeyer; Rott, Marilise Brittes

    2014-12-01

    Acanthamoeba polyphaga is a free-living protozoan pathogen, whose infective trophozoite form is capable of causing a blinding keratitis and fatal granulomatous encephalitis in humans. The damage caused by A. polyphaga trophozoites in human corneal or brain infections is the result of several different pathogenic mechanisms that have not yet been elucidated at the molecular level. We performed a comprehensive analysis of the proteins expressed by A. polyphaga trophozoites, based on complementary 2-DE MS/MS and gel-free LC-MS/MS approaches. Overall, 202 non-redundant proteins were identified. An A. polyphaga proteomic map in the pH range 3-10 was produced, with protein identification for 184 of 370 resolved spots, corresponding to 142 proteins. Additionally, 94 proteins were identified by gel-free LC-MS/MS. Functional classification revealed several proteins with potential importance for pathogen survival and infection of mammalian hosts, including surface proteins and proteins related to defense mechanisms. Our study provided the first comprehensive proteomic survey of the trophozoite infective stage of an Acanthamoeba species, and established foundations for prospective, comparative and functional studies of proteins involved in mechanisms of survival, development, and pathogenicity in A. polyphaga and other pathogenic amoebae.

  6. [Amoebic meningoencephalitis by "Eaegleria" and "Acanthamoeba" (author's transl)].

    PubMed

    de Carneri, I

    1977-01-01

    Primary amoebic meningoencephalitis (ME) by Naegleria fowleri a free-living protozoon found in fresh, warm waters, is a well known fatal disease lasting less than one week. It affects sporadically swimmers and children playing in mud puddles. Less than 100 cases have been described. Recently a more rare, distinct amoebic meningoencephalitis due to some species of free-living Acanthamoeba was identified, lasting some weeks or more but still with a fatal evolution. In this case the amoebae do not always enter the brain directly through the cribrous membranes but cause mild, primary infections of respiratory airways: exceptionally, mainly in immunodepressed subjects, they then reach the CNS causing a secondary ME. Naegleria is fairly sensitive in vitro to some drugs, but in vivo their efficacy is dramtically lowered for pharmacokinetic reasons. Acanthamoeba is in every respect less sensitive. Prophylaxis is almost impossible to achieve. Some diagnostic procedures are described and the importance of their use in diagnosis of the so called aseptic purulent ME is stressed.

  7. Encystation in Acanthamoeba castellanii: development of biocide resistance.

    PubMed

    Lloyd, D; Turner, N A; Khunkitti, W; Hann, A C; Furr, J R; Russell, A D

    2001-01-01

    Since the early 1960s, axenic culture and the development of procedures for the induction of encystation have made Acanthamoeba spp. superb experimental systems for studies of cell biology and differentiation. More recently, since their roles as human pathogens causing keratitis and encephalitis have become widely recognized, it has become urgent to understand the parameters that determine differentiation, as cysts are much more resistant to biocides than are the trophozoites. Viability of trophozoites of the soil amoeba Acanthamoeba castellanii (Neff), is conveniently measured by its ability to form plaques on a lawn of Escherichia coli. Use of confocal laser scanning microscopy with Calcofluor white, Congo Red or the anionic oxonol dye, DiBAC4(3) or flow cytometry with propidium iodide diacetate and fluorescein or oxonol provides more rapid assessment. For cysts, the plaque method is still the best, because dye exclusion does not necessarily indicate viability and therefore the plate count method has been used to study the sequence of development of biocide resistance during the differentiation process. After two hours, resistance to HCl was apparent. Polyhexamethylene biguanide, benzalkonium chloride, propamidine isethionate, pentamidine isethionate, dibromopropamine isethionate, and H2O2 and moist heat, all lost effectiveness at between 14 and 24 h after trophozoites were inoculated into encystation media. Chlorhexidine diacetate resistance was observed at between 24 and 36 h. The molecular biology and biochemistry of the modifications that underlie these changes are now being investigated.

  8. Methanolic extract of Peganum harmala exhibit potent activity against Acanthamoeba castellanii cysts and its encystment in vitro.

    PubMed

    Shohaib, Hafiz Muhammad; Nawaz, Salik; Matin, Abdul

    2016-11-01

    Acanthamoeba castellanii is member of free living amoeba that may cause painful sight-threatening keratitis and life threatening encephalitis which involves central nervous system. Treatments for both infections are problematic because of the amoebic cysts resistance to therapeutic agents. Here we evaluated in vitro strength of methanolic seed extract of Peganum harmala on Acanthamoeba cysts and its encystment mechanism. Our results revealed seed extracts (1 to 30mg/ml) exhibited amoebicidal effects against Acanthamoeba cysts. Furthermore Acanthamoeba encystment was also inhibited in concentration dependent manner with maximum inhibition at 2µg/ml after 48h incubation. In conclusion, we demonstrated for the first time that methanolic extracts exhibit remarkable inhibition of Acanthamoeba cysts and encystment in vitro which could serve a potential new natural agent against Acanthamoeba.

  9. Azole Antifungal Agents To Treat the Human Pathogens Acanthamoeba castellanii and Acanthamoeba polyphaga through Inhibition of Sterol 14α-Demethylase (CYP51).

    PubMed

    Lamb, David C; Warrilow, Andrew G S; Rolley, Nicola J; Parker, Josie E; Nes, W David; Smith, Stephen N; Kelly, Diane E; Kelly, Steven L

    2015-08-01

    In this study, we investigate the amebicidal activities of the pharmaceutical triazole CYP51 inhibitors fluconazole, itraconazole, and voriconazole against Acanthamoeba castellanii and Acanthamoeba polyphaga and assess their potential as therapeutic agents against Acanthamoeba infections in humans. Amebicidal activities of the triazoles were assessed by in vitro minimum inhibition concentration (MIC) determinations using trophozoites of A. castellanii and A. polyphaga. In addition, triazole effectiveness was assessed by ligand binding studies and inhibition of CYP51 activity of purified A. castellanii CYP51 (AcCYP51) that was heterologously expressed in Escherichia coli. Itraconazole and voriconazole bound tightly to AcCYP51 (dissociation constant [Kd] of 10 and 13 nM), whereas fluconazole bound weakly (Kd of 2,137 nM). Both itraconazole and voriconazole were confirmed to be strong inhibitors of AcCYP51 activity (50% inhibitory concentrations [IC50] of 0.23 and 0.39 μM), whereas inhibition by fluconazole was weak (IC50, 30 μM). However, itraconazole was 8- to 16-fold less effective (MIC, 16 mg/liter) at inhibiting A. polyphaga and A. castellanii cell proliferation than voriconazole (MIC, 1 to 2 mg/liter), while fluconazole did not inhibit Acanthamoeba cell division (MIC, >64 mg/liter) in vitro. Voriconazole was an effective inhibitor of trophozoite proliferation for A. castellanii and A. polyphaga; therefore, it should be evaluated in trials versus itraconazole for controlling Acanthamoeba infections.

  10. Azole Antifungal Agents To Treat the Human Pathogens Acanthamoeba castellanii and Acanthamoeba polyphaga through Inhibition of Sterol 14α-Demethylase (CYP51)

    PubMed Central

    Lamb, David C.; Warrilow, Andrew G. S.; Rolley, Nicola J.; Parker, Josie E.; Nes, W. David; Smith, Stephen N.; Kelly, Diane E.

    2015-01-01

    In this study, we investigate the amebicidal activities of the pharmaceutical triazole CYP51 inhibitors fluconazole, itraconazole, and voriconazole against Acanthamoeba castellanii and Acanthamoeba polyphaga and assess their potential as therapeutic agents against Acanthamoeba infections in humans. Amebicidal activities of the triazoles were assessed by in vitro minimum inhibition concentration (MIC) determinations using trophozoites of A. castellanii and A. polyphaga. In addition, triazole effectiveness was assessed by ligand binding studies and inhibition of CYP51 activity of purified A. castellanii CYP51 (AcCYP51) that was heterologously expressed in Escherichia coli. Itraconazole and voriconazole bound tightly to AcCYP51 (dissociation constant [Kd] of 10 and 13 nM), whereas fluconazole bound weakly (Kd of 2,137 nM). Both itraconazole and voriconazole were confirmed to be strong inhibitors of AcCYP51 activity (50% inhibitory concentrations [IC50] of 0.23 and 0.39 μM), whereas inhibition by fluconazole was weak (IC50, 30 μM). However, itraconazole was 8- to 16-fold less effective (MIC, 16 mg/liter) at inhibiting A. polyphaga and A. castellanii cell proliferation than voriconazole (MIC, 1 to 2 mg/liter), while fluconazole did not inhibit Acanthamoeba cell division (MIC, >64 mg/liter) in vitro. Voriconazole was an effective inhibitor of trophozoite proliferation for A. castellanii and A. polyphaga; therefore, it should be evaluated in trials versus itraconazole for controlling Acanthamoeba infections. PMID:26014948

  11. Isolation of Acanthamoeba species in surface waters of Gilan province-north of Iran.

    PubMed

    Mahmoudi, Mohammad Reza; Taghipour, Niloofar; Eftekhar, Mohammad; Haghighi, Ali; Karanis, Panagiotis

    2012-01-01

    We analyzed water samples to determine the prevalence of free-living Acanthamoeba in water sources from Gilan, greater area, Iran. A total of 27 surface water samples were collected from environmental sources, including natural (rivers, lakes, springs, and lagoon) and freshwater source. The samples were filtrated and transferred to non-nutrient agar plates seeded with Escherichia coli and incubated for 2 to 7 days at 30°C or 42°C. The plates were examined by microscopy to morphologically identify Acanthamoeba species. Following DNA extraction, PCR was used to confirm the microscopically identification. A total of 19 out of 27 samples (70.3%) were positive for Acanthamoeba species based on the morphological criteria, and 14 (73.7%) were confirmed by PCR method. The high frequency of Acanthamoeba spp. in different environmental water sources of Gilan is an alert for the public health related to water sources in Iran.

  12. Acanthamoeba keratitis in a non-contact lens wearer with human immunodeficiency virus.

    PubMed

    Hansen, Birgitte; Kronborg, Gitte

    2003-01-01

    Acanthamoeba keratitis is potentially blinding and often associated with contact lens wearing. A human immunodeficiency virus (HIV)-positive patient, a non-contact lens wearer, presented with keratitis. She experienced a protracted course of disease, characterized by exacerbations and remissions, and was treated with various topical antibiotics and steroids. 13 months after symptom onset the eye was removed owing to serious scarring of cornea and unbearable pain. Microbiological and histopathological examination of the cornea showed Acanthamoeba. In non-contact lens wearers suffering from Acanthamoeba keratitis the diagnosis is delayed, pathognomonic features are often not seen and visual outcome is usually poor. There is no known relation between HIV infection and Acanthamoeba keratitis.

  13. Identification of 18S ribosomal DNA genotype of Acanthamoeba from hot spring recreation areas in the central range, Taiwan

    NASA Astrophysics Data System (ADS)

    Hsu, Bing-Mu; Ma, Po-Hua; Liou, Tai-Sheng; Chen, Jung-Sheng; Shih, Feng-Cheng

    2009-04-01

    SummaryAcanthamoeba is a free-living amoebae ubiquitous to aquatic environments. Within the genus a few species are recognized as opportunistic potential human pathogens, which cause granulomatous amoebic encephalitis (GAE) and keratitis. Infections of keratitis are frequently reported through wearing lens while swimming in the non-disinfected aquatic environment. Contaminations in hot tubs, spas and public baths are also possible. As a result, in this study, we identified Acanthamoeba based on the PCR amplification with a genus-specific primer pair and investigated the distribution of Acanthamoeba at five hot spring recreation areas in central range, Taiwan. We gathered data on factors potentially associated with the pathogen's distribution, including various sampling sites, aquatic environment, physical and microbiological water quality parameters. Spring water was collected from 55 sites and Acanthamoeba was detected in 9 (16.4%). The most frequently detected was Acanthamoeba griffini, followed by Acanthamoeba jacobsi. Legionella were detected in 18 (32.7%) of the sites sampled in this study. The species of Legionella identified included Legionella pneumophila serotype 6, serotype 1, and Legionella erythra. Overall, 9.1% of the samples contained both Acanthamoeba and Legionella. The prevalence of Acanthamoeba was contrary to the levels of microbiological indicators recommended by Taiwan CDC, and no significant differences (Mann-Whitney U test, P < 0.05) were observed between the presence/absence of Acanthamoeba and water quality parameters. Results of this survey confirm the existence of Acanthamoeba in Taiwan spring recreation areas. Acanthamoeba, the organism responsible for the majority of Acanthamoeba keratitis and can serve as vehicles for facultative pathogens, should be considered a potential threat for health associated with human activities in spring recreation areas of Taiwan.

  14. [Acanthamoeba sp.: a case report in a non-contact lens wearer].

    PubMed

    Menghi, Claudia; Caride, María C; Gatta, Claudia

    2012-01-01

    Acanthamoeba sp. keratitis is generally related to wearing contact lenses, and, to a lesser extent, to contaminated water. It is characterized by reduced visual capacity and the presence of severe ocular pain. Clinically, it can be mistaken for a herpes infection. if it is not diagnosed, and timely and adequately treated, it can result in corneal perforation and, eventually, in vision loss. One of the few registered cases of Acanthamoeba sp. keratitis not related to the use of contact lenses is herein reported.

  15. Reevaluation of an Acanthamoeba Molecular Diagnostic Algorithm following an Atypical Case of Amoebic Keratitis.

    PubMed

    Lau, Rachel; Cunanan, Marlou; Jackson, Jonathan; Ali, Ibne Karim M; Chong-Kit, Ann; Gasgas, Jason; Tian, Jinfang; Ralevski, Filip; Boggild, Andrea K

    2015-10-01

    Amoebic keratitis (AK) is a potentially blinding infection, the prompt diagnosis of which is essential for limiting ocular morbidity. We undertook a quality improvement initiative with respect to the molecular detection of acanthamoebae in our laboratory because of an unusual case of discordance. Nine ATCC strains of Acanthamoeba and 40 delinked, biobanked, surplus corneal scraping specimens were analyzed for the presence of acanthamoebae with four separate real-time PCR assays. The assay used by the Free-Living and Intestinal Amebas Laboratory of the CDC was considered the reference standard, and the performance characteristics of each individual assay and pairs of assays were calculated. Outcome measures were sensitivity, specificity, positive predictive value (PPV), and negative predictive value (NPV). Of 49 included specimens, 14 (28.6%) were positive by the gold standard assay, and 35 (71.4%) were negative. The sensitivities of the individual assays ranged from 64.3% to 92.9%, compared to the gold standard, while the specificities ranged from 88.6% to 91.4%. The PPVs and NPVs ranged from 69.2% to 78.6% and from 86.1% to 96.9%, respectively. Combinations of assay pairs led to improved performance, with sensitivities ranging from 92.9% to 100% and specificities ranging from 97.1% to 100%. ATCC and clinical strains of Acanthamoeba that failed to be detected by certain individual assays included Acanthamoeba castellanii, Acanthamoeba culbertsoni, and Acanthamoeba lenticulata. For three clinical specimens, false negativity of the gold standard assay could not be excluded. Molecular diagnostic approaches, especially combinations of highly sensitive and specific assays, offer a reasonably performing, operator-independent, rapid strategy for the detection of acanthamoebae in clinical specimens and are likely to be more practical than either culture or direct microscopic detection.

  16. Vectorial role of Acanthamoeba in Legionella propagation in water for human use.

    PubMed

    Magnet, A; Peralta, R H S; Gomes, T S; Izquierdo, F; Fernandez-Vadillo, C; Galvan, A L; Pozuelo, M J; Pelaz, C; Fenoy, S; Del Águila, C

    2015-02-01

    Legionella spp. is the causative agent of Legionnaires' disease and is transmitted through aerosols emanating from man-made water systems. Legionella resistance to water treatments has been related to its association with environmental amoebae such as Acanthamoeba. Due to the high presence of this protozoon in Spain and the high rate of notification of Legionnaires' disease of this country, the aims of this work were to study the coexistence of these bacteria and protozoa in water as well as their interaction. The usefulness of Acanthamoeba co-culture for the isolation of environmental Legionella was also studied. For this purpose, 70 water samples were collected in 2011 from three Drinking Water Treatment Plants, three Wastewater Treatment Plants and five Natural Pools in Spain. Acanthamoeba was found by PCR in 87.1% (61/70) samples and, by culture in 85.7% (60/70) samples. Legionella was detected by PCR in 58.6% (41/70) of water samples, in 5.7% (4/70) by agar culture and 75.7% (53/70) by Acanthamoeba co-culture. From the 54 Acanthamoeba water isolates, Legionella was detected in 43 of them independently of Acanthamoeba's genotype (T3, T4 and T11). Legionella feeleii, Legionella birminghamiensis, Legionella gresilensis/berliardensis, Legionella fairfieldensis, Legionella drozanski and Legionella falloni were identified. In conclusion, our results showed that environmental Acanthamoeba is infected by Legionella to a high percentage, and due to its ubiquity, high resistance and its pathogenic potential per se, new methods for its elimination should be studied. Also, the high effectivity of Acanthamoeba co-culture for Legionella detection has been shown.

  17. Reevaluation of an Acanthamoeba Molecular Diagnostic Algorithm following an Atypical Case of Amoebic Keratitis

    PubMed Central

    Lau, Rachel; Cunanan, Marlou; Jackson, Jonathan; Ali, Ibne Karim M.; Chong-Kit, Ann; Gasgas, Jason; Tian, Jinfang; Ralevski, Filip

    2015-01-01

    Amoebic keratitis (AK) is a potentially blinding infection, the prompt diagnosis of which is essential for limiting ocular morbidity. We undertook a quality improvement initiative with respect to the molecular detection of acanthamoebae in our laboratory because of an unusual case of discordance. Nine ATCC strains of Acanthamoeba and 40 delinked, biobanked, surplus corneal scraping specimens were analyzed for the presence of acanthamoebae with four separate real-time PCR assays. The assay used by the Free-Living and Intestinal Amebas Laboratory of the CDC was considered the reference standard, and the performance characteristics of each individual assay and pairs of assays were calculated. Outcome measures were sensitivity, specificity, positive predictive value (PPV), and negative predictive value (NPV). Of 49 included specimens, 14 (28.6%) were positive by the gold standard assay, and 35 (71.4%) were negative. The sensitivities of the individual assays ranged from 64.3% to 92.9%, compared to the gold standard, while the specificities ranged from 88.6% to 91.4%. The PPVs and NPVs ranged from 69.2% to 78.6% and from 86.1% to 96.9%, respectively. Combinations of assay pairs led to improved performance, with sensitivities ranging from 92.9% to 100% and specificities ranging from 97.1% to 100%. ATCC and clinical strains of Acanthamoeba that failed to be detected by certain individual assays included Acanthamoeba castellanii, Acanthamoeba culbertsoni, and Acanthamoeba lenticulata. For three clinical specimens, false negativity of the gold standard assay could not be excluded. Molecular diagnostic approaches, especially combinations of highly sensitive and specific assays, offer a reasonably performing, operator-independent, rapid strategy for the detection of acanthamoebae in clinical specimens and are likely to be more practical than either culture or direct microscopic detection. PMID:26202123

  18. In the case of transmission of Mycobacterium ulcerans in buruli ulcer disease Acanthamoeba species stand accused.

    PubMed

    Wilson, M D; Boakye, D A; Mosi, L; Asiedu, K

    2011-03-01

    Buruli ulcer disease caused by Mycobacterium ulcerans results in extensive destruction of skin and soft tissue and long-term functional disabilities that ultimately require surgery and rehabilitation. The disease is associated with aquatic and swampy environments with the mycobacterium occurring in biofilms, soil, aquatic insects, fish and wildlife however, the mode of transmission to humans remains an enigma. Current transmission ideas including bites from predatory water bugs and mosquitoes, do not explain satisfactorily the spasmodic disease distribution in human populations. Here we argue that Acanthamoeba species are the natural hosts of M. ulcerans and are mainly responsible for disease transmission because; (i) Acanthamoebae are known natural hosts of several microbial pathogens including M. marinum, M. avium and Legionella pneumophila, (ii) culture of slow-to-grow microbial pathogens hosted in nature by Acanthamoeba spp is enhanced when the media is seeded with the protozoa, (iii) acanthamoebae and M. ulcerans share similar bio-ecological and epidemiological settings, (iv) documented evidence that prior growth of L. pneumophila and M. avium in acanthamoebae influences entry mechanisms, intracellular growth and virulence in human monocytes, (v) Acanthamoeba spp also infect humans and cause diseases via routes of openings including broken skin and sites of trauma similar to M. ulcerans and (vi) M. ulcerans is rather a fastidious intracellular organism as recent analysis of the genome indicate. We argue further that temperature plays a significant role in transmission determining the fate of either the intracellular microbe or the host cells. Also, Acanthamoeba-pathogen association has a long evolutionary history because the same set of bacterial genes and gene products e.g. in L. pneumophila are required for survival in both mammalian and protozoan host cells. We suggest that the involvement of Acanthamoeba in the transmission of M. ulcerans to humans better

  19. Standardized Method of Measuring Acanthamoeba Antibodies in Sera from Healthy Human Subjects

    PubMed Central

    Chappell, Cynthia L.; Wright, John A.; Coletta, Michael; Newsome, Anthony L.

    2001-01-01

    Acanthamoeba species can cause serious, debilitating, and sometimes life-threatening infections. Three groups have been identified using morphological and immunological comparisons. Previous serological studies have utilized a variety of antigen preparations and assay methods and reported disparate (3 to 100%) results. This study was designed to (i) optimize an enzyme-linked immunosorbent assay for detecting serum antibodies to each of the Acanthamoeba serogroups and (ii) test 55 healthy individuals for specific immunoglobulin G reactivity. The highest signal-to-background ratio was found when 3,000 fixed, intact trophozoites per well were used with a 1:10 serum dilution. Sera yielding optical densities of <0.25 against all three Acanthamoeba serogroups were used to define the cutoff for positive results. The highest background reactivity with these sera was seen with Acanthamoeba polyphaga (serogroup 2), followed by Acanthamoeba culbertsoni (serogroup 3) and Acanthamoeba astronyxis (serogroup 1). Of 55 subjects tested, the highest number of positive results was seen with A. polyphaga (81.8%), followed by A. astronyxis (52.8%) and A. culbertsoni (40%). Seven serum samples (12.7%) were negative for all three Acanthamoeba serogroups, 16 (29.1%) were positive for one serogroup only, 16 were positive for two serogroups, and 16 reacted to all three serogroups. Further analysis showed no significant associations between serogroup reactivity and age or gender. However, some ethnic differences were noted, especially with A. polyphaga antigens. In that case, serum samples from Hispanic subjects were 14.5 times less likely to be positive (P = 0.0025) and had lower mean absorbance values (P = 0.047) than those from Caucasian subjects. Overall, these data suggest that Acanthamoeba colonization or infection is more common than previously thought. Mild or asymptomatic infections may contribute to the observed serum reactivities. PMID:11427418

  20. Cloning and characterization of a novel mannose-binding protein of Acanthamoeba.

    PubMed

    Garate, Marco; Cao, Zhiyi; Bateman, Erik; Panjwani, Noorjahan

    2004-07-09

    Acanthamoebae produce a painful, blinding infection of the cornea. The mannose-binding protein (MBP) of Acanthamoeba is thought to play a key role in the pathogenesis of the infection by mediating the adhesion of parasites to the host cells. We describe here the isolation and molecular cloning of Acanthamoeba MBP. The MBP was isolated by chromatography on the mannose affinity gel. Gel filtration experiments revealed that the Acanthamoeba lectin is a approximately 400-kDa protein that is constituted of multiple 130-kDa subunits. Cloning and sequencing experiments indicated that the Acanthamoeba MBP gene is composed of 6 exons and 5 introns that span 3.6 kb of the amoeba genome and that MBP cDNA codes for a precursor protein of 833 amino acids. That the cloned cDNA encodes authentic MBP was demonstrated by showing that: (i). recombinant MBP possesses mannose binding activity, and (ii). polyclonal antibodies prepared against Acanthamoeba MBP bound to the recombinant protein. Sequence analysis revealed that the MBP contains a large N-terminal extracellular domain, a transmembrane domain, and a short C-terminal cytoplasmic domain. Despite extensive BLAST searches using the MBP sequence, no significant matches were retrieved. The most striking feature of the Acanthamoeba MBP sequence is the presence of a cysteine-rich region containing 14 CXCXC motifs within the extracellular domain. In summary, we have isolated, cloned, and characterized a novel MBP from Acanthamoeba. Because the presence of antibodies to MBP in tears provides protection against infection, the availability of the MBP cDNA sequence and rMBP should help develop: (i). a tear-based test to identify individuals who are at risk of developing the keratitis and (ii). strategies to immunize high-risk individuals.

  1. Higher-order aberrations and best-corrected visual acuity in Native American children with a high prevalence of astigmatism

    PubMed Central

    Miller, Joseph M.; Harvey, Erin M.; Schwiegerling, Jim

    2016-01-01

    Purpose To determine whether higher-order aberrations (HOAs) in children from a highly astigmatic population differ from population norms and whether HOAs are associated with astigmatism and reduced best-corrected visual acuity. Methods Subjects were 218 Tohono O’odham Native American children 5–9 years of age. Noncycloplegic HOA measurements were obtained with a handheld Shack-Hartmann sensor (SHS). Signed (z06s to z14s) and unsigned (z06u to z14u) wavefront aberration Zernike coefficients Z(3,−3) to Z(4,4) were rescaled for a 4 mm diameter pupil and compared to adult population norms. Cycloplegic refraction and best-corrected logMAR letter visual acuity (BCVA) were also measured. Regression analyses assessed the contribution of astigmatism (J0) and HOAs to BCVA. Results The mean root-mean-square (RMS) HOA of 0.191 ± 0.072 μm was significantly greater than population norms (0.100 ± 0.044 μm. All unsigned HOA coefficients (z06u to z14u) and all signed coefficients except z09s, z10s, and z11s were significantly larger than population norms. Decreased BCVA was associated with astigmatism (J0) and spherical aberration (z12u) but not RMS coma, with the effect of J0 about 4 times as great as z12u. Conclusions Tohono O’odham children show elevated HOAs compared to population norms. Astigmatism and unsigned spherical aberration are associated with decreased acuity, but the effects of spherical aberration are minimal and not clinically significant. PMID:26239206

  2. Resistance of Acanthamoeba cysts to disinfection in multiple contact lens solutions.

    PubMed

    Johnston, Stephanie P; Sriram, Rama; Qvarnstrom, Yvonne; Roy, Sharon; Verani, Jennifer; Yoder, Jonathan; Lorick, Suchita; Roberts, Jacquelin; Beach, Michael J; Visvesvara, Govinda

    2009-07-01

    Acanthamoebae are free-living amoebae found in the environment, including soil, freshwater, brackish water, seawater, hot tubs, and Jacuzzis. Acanthamoeba species can cause keratitis, a painful vision-threatening infection of the cornea, and fatal granulomatous encephalitis in humans. More than 20 species of Acanthamoeba belonging to morphological groups I, II, and III distributed in 15 genotypes have been described. Among these, Acanthamoeba castellanii, A. polyphaga, and A. hatchetti are frequently identified as causing Acanthamoeba keratitis (AK). Improper contact lens care and contact with nonsterile water while wearing contact lenses are known risk factors for AK. During a recent multistate outbreak, AK was found to be associated with the use of Advanced Medical Optics Complete MoisturePlus multipurpose contact lens solution, which was hypothesized to have had insufficient anti-Acanthamoeba activity. As part of the investigation of that outbreak, we compared the efficacies of 11 different contact lens solutions against cysts of A. castellanii, A. polyphaga, and A. hatchetti (the isolates of all species were genotype T4), which were isolated in 2007 from specimens obtained during the outbreak investigation. The data, generated with A. castellanii, A. polyphaga, and A. hatchetti cysts, suggest that the two contact lens solutions containing hydrogen peroxide were the only solutions that showed any disinfection ability, with 0% and 66% growth, respectively, being detected with A. castellanii and 0% and 33% growth, respectively, being detected with A. polyphaga. There was no statistically significant difference in disinfection efficacy between the 11 solutions for A. hatchetti.

  3. Comparative analyses of different genetic markers for the detection of Acanthamoeba spp. isolates.

    PubMed

    Derda, Monika; Wojtkowiak-Giera, Agnieszka; Hadaś, Edward

    2014-09-01

    Acanthamoeba are widespread free-living amoebae which may cause granulomatous amoebic encephalitis (GAE), keratitis, skin ulcerations and disseminated tissue infection. An important diagnostic and prognostic factor for the treatment of infection is a quick and correct diagnosis of amoebae strains. The aim of our study was to develop a rapid method for detection and identification of pathogenic Acanthamoeba spp. strains from diagnostic material collected from water. In this study we analysed five amplification-based genetic markers (Aca 16S, Ac6/210, GP, JDP, Nelson) used for identification of pathogenic Acanthamoeba spp. strains isolated in water sources in Poland, Iceland and Sweden. Our results demonstrated the presence of pathogenic Acanthamoeba strains in tap water. PCR assay appeared to be a more rapid and sensitive method to detect the presence of amoebae than the limited conventional techniques. Based on our observations, we can confirm that the use of four out of five genetic markers (Aca 16S, Ac 6/210, JDP, GP, Nelson) may be helpful in identification of Acanthamoeba spp. strains, but only one Aca 16S primer pair is a highly specific marker that distinguishes between pathogenic strains of Acanthamoeba and other free-living amoeba families.

  4. Isolation and characterization of Acanthamoeba strains from soil samples in Gran Canaria, Canary Islands, Spain.

    PubMed

    Reyes-Batlle, María; Todd, Cheridah D; Martín-Navarro, Carmen M; López-Arencibia, Atteneri; Cabello-Vilchez, Alfonso Martín; González, Ana C; Córdoba-Lanús, Elizabeth; Lindo, John F; Valladares, Basilio; Piñero, José E; Lorenzo-Morales, Jacob

    2014-04-01

    Free-living Amoebae of Acanthamoeba genus include non-pathogenic and pathogenic strains that are currently classified in 18 different genotypes, T1-T18. In this study, a survey was carried out to evaluate the presence of Acanthamoeba strains in soil samples collected between 2012 and 2013 in Gran Canaria Island, Canary Islands, Spain. Samples were inoculated onto non-nutrient agar (NNA) plates and were checked for the presence of Acanthamoeba. Identification of Acanthamoeba strains was based on the morphology of the cyst and trophozoite forms. Subsequently, positive samples were cloned for their molecular characterization at the genotype level by sequencing the DF3 region located in the 18S rDNA gene of Acanthamoeba as previously described. Sequencing results revealed the presence of T2, T5 and T4 genotypes within the studied samples. To the best of our knowledge, this is the first report demonstrating the presence of Acanthamoeba in Gran Canaria Island and the first study at the genotype level in the Canary Islands.

  5. Evaluation of Ozone Application in Dental Unit Water Lines Contaminated with Pathogenic Acanthamoeba

    PubMed Central

    HIKAL, Wafaa; ZAKI, Basma; SABRY, Hany

    2015-01-01

    Background: In this study morphological and molecular characterization of Acanthamoeba strains, isolated from dental unit waterlines (DUWLs) were surveyed and the levels of disinfection achievable in vitro by the application of ozone disinfectant to DUWLs were evaluate. Methods: Water samples were collected from air-water syringes, cup fillers and tap water before and at the end of the working day. They were cultured on non-nutrient agar (NNA) plates. Species identification was carried out with a PCR assay based on sequence analysis of the 18S rRNA gene. The cellular response to ozone was tested on Acanthamoeba cyst with different doses at different contact time in vitro twice. Results: Prevalence rates for Acanthamoeba contamination were 100, 100 and 72% for air-water syringes, cup fillers and tap water, respectively. The morphological analysis revealed the presence of A. castellanii, A. griffin, A. hatchitti and A. lenticulata. Phylogenetic analysis of the sequences showed the four strains to be closely related to a sequence type (T3, T4, T5 and T11). Acanthamoeba cells were stained with trypan blue, which revealed killed of Acanthamoeba instantaneously after 10 minutes in ozonized water. There was no growth of Acanthamoeba occurred after ozone treatment in water bottles for 5 minutes with a flow rate of 500 mg/hour. Conclusion : Ozone can play an important role in controlling the problem of contamination of DUWLs as a potent disinfectant. PMID:26622296

  6. Detection of Acanthamoeba and Toxoplasma in River Water Samples by Molecular Methods in Iran

    PubMed Central

    MAHMOUDI, Mohammad Reza; KAZEMI, Bahram; HAGHIGHI, Ali; KARANIS, Panagiotis

    2015-01-01

    Background: Free-living amoebae such as Acanthamoeba species may act as carriers of Cryptosporidium and Toxoplasma oocysts, thus, may play an important role in the water-borne transmission of these parasites. In the present study, a loop mediated isothermal amplification (LAMP) method for detection of Toxoplasma and a PCR assay were developed for investigation of Acanthamoeba in environmental water samples. Methods: A total of 34 samples were collected from the surface water in Guilan Province. Water samples were filtrated with membrane filters and followed by DNA extraction. PCR and LAMP methods used for detection of the protozoan parasites Acanthamoeba and Toxoplasma respectively. Results: Totally 30 and 2 of 34 samples were positive for Acanthamoeba and Toxoplasma oocysts respectively. Two samples were positive for both investigated parasites. Conclusion: The investigated water supplies, are contaminated by Toxoplasma and Acanthamoeba (oo)cystes. Acanthamoeba may play an important role in water-borne transmission of Toxoplasma in the study area. For the first time in Iran, protocol of LAMP method was used effectively for the detection of Toxoplasma in surface water samples in Iran. PMID:26246823

  7. Health effects of Acanthamoeba spp. and its potential for waterborne transmission.

    PubMed

    Nwachuku, Nena; Gerba, Charles P

    2004-01-01

    Risk from Acanthamoeba keratitis is complex, depending upon the virulence of the particular strain, exposure, trauma, or other stress to the eye, and host immune response. Bacterial endosymbionts may also play a factor in the pathogenicity of Acanthamoeba. Which factor(s) may be the most important is not clear. The ability of the host to produce IgA antibodies in tears may be a significant factor. The immune response of the host is a significant risk factor for GAE infection. If so, then a certain subpopulation with an inability to produce IgA in the tears may be at greatest risk. There was no sufficient data on the occurrence or types of Acanthamoeba in tapwater in the U.S. Published work on amoebal presence in tapwater does not provide information on the type of treatment the water received or the level of residual chlorine. Assessment of the pathogenicity by cell culture and molecular methods of Acanthamoeba in tapwater would also be useful in the risk assessment process for drinking water. The possibility that Acanthamoeba spp. might serve as vectors for bacterial infections from water sources also should be explored. The bacterial endosymbionts include an interesting array of pathogens such as Vibrio cholerae and Legionella pneumophila, both of which are well recognized waterborne/water-based pathogens. Work is needed to determine if control of Acanthamoeba spp. is needed to control water-based pathogens in water supplies.

  8. First report of an Acanthamoeba genotype T13 isolate as etiological agent of a keratitis in humans.

    PubMed

    Grün, Anna-Lena; Stemplewitz, Birthe; Scheid, Patrick

    2014-06-01

    Several strains of free-living amoebae (FLA) belonging to the genus Acanthamoeba are able to cause a painful sight-threatening disease of the cornea designated as Acanthamoeba keratitis (AK). In this case report, a 22-year-old woman, wearer of soft contact lenses, was treated after the initial examination, and follow-up laboratory results led to the diagnosis of Acanthamoeba keratitis. The patient recovered under the targeted therapy, demonstrating that the acanthamoebae were the etiological agents of the keratitis in this case. The acanthamoebae belonged morphologically to group II. Genotyping of the causative Acanthamoeba strain based on sequences of the PCR amplimer ASA.S1 amplified from 18S ribosomal DNA by using the genus-specific primers JDP1 and JDP2 followed. The phylogenetic comparison of ASA.S1 confirmed that the isolated Acanthamoeba strain is closely related to genotype T13 supported by pairwise sequence identities of 97.1-98.0% and bootstrap support of 980 replicates with reference sequences of genotype T13. These results regarding the Acanthamoeba keratitis-causing isolate KaBo expands the number of known pathogenic genotypes to 12. To our knowledge, this is the first report of a T13 Acanthamoeba genotype being associated with keratitis in humans.

  9. Comparison of PCR, microscopic examination and culture for the early diagnosis and characterization of Acanthamoeba isolates from ocular infections.

    PubMed

    Yera, H; Zamfir, O; Bourcier, T; Ancelle, T; Batellier, L; Dupouy-Camet, J; Chaumeil, C

    2007-03-01

    In the study presented here, PCR, microscopic examination and culture of corneal samples were compared as methods of confirming the clinical diagnosis of Acanthamoeba keratitis, a serious ocular infection that is difficult to diagnose and threatens eyesight. The three methods were applied to isolates obtained from 513 patients with clinical signs or risk factors suggesting Acanthamoeba infection. Acanthamoeba keratitis was diagnosed in 12 of these patients. Combined PCR assays were more sensitive (94%) than either microscopic examination (33%) or culture (7%). The Acanthamoeba isolates were characterized using DNA sequence analysis of the nuclear small-subunit rRNA gene, and T4 was the predominant genotype found.

  10. Characterisation and differentiation of pathogenic and non-pathogenic Acanthamoeba strains by their protein and antigen profiles.

    PubMed

    Walochnik, J; Sommer, K; Obwaller, A; Haller-Schober, E-M; Aspöck, H

    2004-03-01

    Free-living amoebae of the genus Acanthamoeba are the causative agents of Acanthamoeba keratitis (AK) and granulomatous amoebic encephalitis. Acanthamoebae occur ubiquitously in the environment and are thus a constant cause of antigenic stimulation. In a previous study we have shown that compared to control sera, AK patients exhibit markedly lower immunoreactivities to whole cell antigen of Acanthamoeba spp. As the pathogenicity of acanthamoebae primarily relies on the excretion of proteins, it was the aim of the present study to investigate the immunoreactivity of metabolic antigen from different Acanthamoeba strains of varying pathogenicity. Three Acanthamoeba strains, one highly pathogenic, one non-pathogenic but thermophilic and one non-thermophilic non-pathogenic, were used for antigen extraction. The antigen was harvested before and after contact with human cells and all strains were tested with AK sera and with sera from healthy individuals. It was shown that the somatic protein profiles of the Acanthamoeba strains correlated to the morphological groups, and that within morphological group II-the group associated with AK-the profiles of the metabolic antigens correlated to strain pathogenicity. Moreover, it was shown that the control sera showed markedly higher immunoreactivities than the sera of the AK patients and that this immunoreactivity was generally higher to the non-pathogenic strains than to the pathogenic strain. Altogether our results once again raise the question of whether there is an immunological predisposition in AK. To our knowledge this is the first study on the immunoreactivity of metabolic antigen of acanthamoebae.

  11. Fas deficiency in mice with the Balb/c background induces blepharitis with allergic inflammation and hyper-IgE production in conjunction with severe autoimmune disease.

    PubMed

    Takahashi, Suzuka; Futatsugi-Yumikura, Shizue; Fukuoka, Ayumi; Yoshimoto, Tomohiro; Nakanishi, Kenji; Yonehara, Shin

    2013-05-01

    Fas (CD95) is a cell surface death receptor belonging to the tumor necrosis factor receptor superfamily, which mediates apoptosis-inducing signaling when activated by Fas ligand or its agonistic antibody. lpr mice with a loss of apoptosis-inducing function mutation in the Fas gene develop systemic autoimmune disease and lymphadenopathy but not allergic inflammation. In the case of Fas mutations including lpr and knockout (KO), background genes determine the incidence and severity of lymphadenopathy and histopathological manifestation of systemic autoimmunity: MRL-lpr/lpr mice and C57BL/6-lpr/lpr or C57BL/6 Fas KO mice develop severe and minimum disease, respectively. We generated Fas KO mice with the Balb/c background that show severer autoimmune phenotypes than MRL-lpr/lpr mice, such as critical infiltration of mononuclear cells into lung, liver and spleen, elevated serum levels of auto-antibodies and a decreased life span. To our astonishment, Balb/c Fas KO mice spontaneously develop blepharitis with not only autoimmune inflammation with deposition of auto-antibody but also allergic inflammation with infiltration by eosinophils and mast cells and show the capacity to strongly increase serum level of IgE and IgG1 along with their aging. Thus, Fas expression regulates development of not only autoimmune disease but also allergic inflammation.

  12. Digital in-line holography for the characterization of flowing particles in astigmatic optical systems

    NASA Astrophysics Data System (ADS)

    Sentis, Matthias P. L.; Bruel, Laurent; Charton, Sophie; Onofri, Fabrice R. A.; Lamadie, Fabrice

    2017-01-01

    An extended Generalized Fresnel Transform (GFT) is proposed to account for the astigmatism introduced by optical elements described, in the paraxial approximation, with a ray transfer matrix analysis. Generalized impulse response and generalized Fresnel transfer function propagators as well as sampling conditions are derived to properly implement this transformation. As a test case, the near-field diffraction patterns and in-line holograms produced by droplets flowing in a tube with cylindrical interfaces have been simulated. A best fitting approach is introduced to retrieve, from the propagated holograms, the 3D position and size of the droplets. Several hologram focusing indicators based on the analysis of droplets focus region are also proposed to further improve the estimation of the droplets position along the optical axis. Numerical simulations and experimental results confirm the applicability and accuracy of the proposed methods.

  13. Image digitalization and processing of contact lens fitting to astigmatic eyes

    NASA Astrophysics Data System (ADS)

    Costa, Manuel F. M.

    1998-01-01

    The use of standard CCD cameras and image digitalization and processing on medical diagnosis are more and more frequent. The correction of human eye's refraction problems by the use of contact lenses is generalized. In spite its advantages in terms of users comfort, special care must be taken on its prescription and adaptation. Astigmatic eyes often place the highest problems. A careful assessment of the quality of the lens to cornea adaptation must to be performed. The basic and more traditional way to check the contact lens' fitting is to perform a fluorescein test. We intend to make the process more convenient for both patient and optometrist. The fluorescence images are acquired by a CCD camera and then digitized and processed in order to produce a semi- automated process.

  14. [Acanthamoeba isolated from child thymus: Acanthamoeba growth during cocultivation with human and murine eukaryotic cells and induction of cytopathogenic effect on these cells].

    PubMed

    Komogorova, V V; Sharova, N I; Gordeeva, L M; Iarilin, A A

    2003-01-01

    A cell line of the ameba assigned to the genus Acanthamoeba by morphological and cariotypic signs and by the specific features of its life cycle was isolated from the thymus of a child operated on for heart disease. The line received the name CDHT (Cells Derived from Human Thymus). Actively proliferating mammalian cells maintain the multiplication of Acanthamoeba cells in vitro. Using thymocytes as an example, it was shown that death of these cells occurred through the mechanism of apoptosis. The human thymocytes preincubated with the supernatant obtained through the cocultivation of CDHT and thymocytes may also undergo apoptosis.

  15. Arcobacter butzleri survives within trophozoite of Acanthamoeba castellanii.

    PubMed

    Villanueva, María P; Medina, Gustavo; Fernández, Heriberto

    2016-01-01

    The survival of three Arcobacter butzleri strains inside Acanthamoeba castellanii was assessed using axenic cultures of A. castellanii that were inoculated with the tested strains and incubated at 26°C under aerobic conditions for 240h. The behavior of bacteria in contact with amoebae was monitored using phase contrast microscopy. The bacterial survival rate within amoebae was assessed through counting colony forming units, using the gentamicin protection assay. All A. butzleri strains were able to survive during 240h within the amoebae, thus suggesting that (i) A. butzleri resists the amoebic digestion processes at least for the analyzed time; (ii) that A. castellanii could serve as an environmental reservoir for this bacterium, probably acting as a transmission vehicle for A. butzleri.

  16. Genotyping and phylogenetic analysis of Acanthamoeba isolates associated with keratitis.

    PubMed

    Risler, Arnaud; Coupat-Goutaland, Bénédicte; Pélandakis, Michel

    2013-11-01

    We examined a partial SSU-rDNA sequence from 20 Acanthamoeba isolates associated with keratitis infections. The phylogenetic tree inferred from this partial sequence allowed to assign isolates to genotypes. Among the 20 isolates examined, 16 were found to be of the T4 genotype, 2 were T3, 1 was a T5, and 1 was a T2, confirming the predominance of T4 in infections. However, the study highlighted other genotypes more rarely associated with infections, particularly the T2 genotype. Our study is the second one to detect that this genotype is associated with keratitis. Additionally, the phylogenetic analyses showed five main emerging clusters, T4/T3/T11, T2/T6, T10/T12/T14, T13/T16, and T7/T8/T9/T17, regularly obtained whichever method was used. A similar branching pattern was found when the full rDNA sequence was investigated.

  17. Acanthamoeba castellanii: morphological analysis of the interaction with human cornea.

    PubMed

    Omaña-Molina, Maritza; González-Robles, Arturo; Salazar-Villatoro, Lizbeth Iliana; Cristóbal-Ramos, Ana Ruth; González-Lázaro, Mónica; Salinas-Moreno, Edmundo; Méndez-Cruz, Rene; Sánchez-Cornejo, Manuel; De la Torre-González, Enrique; Martínez-Palomo, Adolfo

    2010-09-01

    The present study demonstrates that when Acanthamoeba castellanii trophozoites are co-cultivated with isolated human corneas, the amoeba can be invasive and cause damage to the intact corneal epithelium without the requirement of previous corneal abrasion. After adhesion, A. castellanii trophozoites migrate between cells forming bumps on the corneal cell layers and reaching Bowman s membrane in 3h, although no evidence of cell damage was observed until the phagocytic process was detected. Likewise, conditioned medium produced damage to the corneal cells that was proportional to the time of incubation, but this cytophatic effect involved only the most superficial layer of the human cornea and was not enough to explain amoebic invasion of Bowman s membrane. As a result of our observations, we suggest that the mechanical action of the trophozoites and phagocytosis of corneal cells during the process of corneal invasion are more important than previously suggested.

  18. Interactions of some common pathogenic bacteria with Acanthamoeba polyphaga.

    PubMed

    Huws, Sharon A; Morley, Robert J; Jones, Martin V; Brown, Michael R W; Smith, Anthony W

    2008-05-01

    Protozoan grazing is a major trophic pathway whereby the biomass re-enters the food web. Nonetheless, not all bacteria are digested by protozoa and the number known to evade digestion, resulting in their environmental augmentation, is increasing. We investigated the interactions of Bacillus cereus, Enterococcus faecalis, Enteropathogenic Escherichia coli (EPEC), Listeria monocytogenes, Salmonella enterica serovar Typhimurium, and methicillin-sensitive Staphylococcus aureus (MSSA), with the amoeba, Acanthamoeba polyphaga. There was evidence of predation of all bacterial species except L. monocytogenes and S. aureus, where extracellular numbers were significantly higher when cultured with amoebae compared with growth in the absence of amoebae. Intracellular growth kinetic experiments and fluorescent confocal microscopy suggest that S. aureus survived and may even multiply within A. polyphaga, whereas there was no apparent intra-amoebal replication of L. monocytogenes and higher numbers were likely sustained on metabolic waste products released during coculture.

  19. Nitrogen Mineralization by Acanthamoeba polyphaga in Grazed Pseudomonas paucimobilis Populations

    PubMed Central

    Sinclair, James L.; McClellan, J. Forbes; Coleman, David C.

    1981-01-01

    Nitrogen mineralization was studied in a simple grazing system in which the protozoan Acanthamoeba polyphaga was grown with the bacterium Pseudomonas paucimobilis (two soil organisms isolated from the shortgrass prairie in northern Colorado). In different experiments, either carbon or nitrogen was adjusted to be in limiting amounts. When carbon was limiting, grazers were almost entirely responsible for nitrogen mineralization, with bacteria themselves contributing little. When nitrogen was limiting, nitrogen mineralization by grazers permitted continued growth by the grazed bacteria and a greater bacterial biomass production. The increased growth of the grazed bacteria did not result in an increased total amount of carbon used, but the grazed bacteria used carbon more efficiently than the ungrazed bacteria. PMID:16345864

  20. Acanthamoeba, bacterial, and fungal contamination of contact lens storage cases.

    PubMed Central

    Gray, T B; Cursons, R T; Sherwan, J F; Rose, P R

    1995-01-01

    BACKGROUND--Microbial corneal infection is the most serious complication of contact lens wear. Contact lens cases are a recognised potential source of pathogens associated with corneal ulcers. METHODS--This survey established the incidence of protozoal, bacterial, and fungal contact lens case contamination in 101 asymptomatic daily wear cosmetic contact lens wearers from a domiciliary contact lens practice. RESULTS--Eighty two (81%) contact lens cases were found to be contaminated, with 19 (19%) sterile. Of all contact lens cases, 78 (77%) grew bacteria, 24 (24%) fungi, and 20 (20%) protozoa. Acanthamoeba spp were isolated from eight (8%) contact lens cases. Fifty six (55%) contact lens cases yielded mixed bacterial contamination. This is the first contact lens case survey in which hydrogen peroxide disinfection was the major method of contact lens disinfection (75% of subjects) and no home made saline was used. All the contaminating organisms were shown to possess the enzyme catalase that breaks down hydrogen peroxide to oxygen and water. The polymicrobial nature of the biofilms found in many contact lens cases is illustrated electron micrographically. CONCLUSION--Based on data from this and previous studies, the authors conclude with recommendations for contact lens wearers: (1) regular scrubbing of contact lens case interior to disrupt biofilms; (2) exposure of contact lens case to very hot water (> or = 70 degrees C) will kill Acanthamoeba contaminants; (3) allow contact lens case to air dry between uses; (4) if hydrogen peroxide disinfection is preferred, use a two step system; (5) replace contact lens case regularly. Images PMID:7626578

  1. Molecular characterization of Acanthamoeba isolated in water treatment plants and comparison with clinical isolates.

    PubMed

    Magnet, A; Galván, A L; Fenoy, S; Izquierdo, F; Rueda, C; Fernandez Vadillo, C; Pérez-Irezábal, J; Bandyopadhyay, K; Visvesvara, G S; da Silva, A J; del Aguila, C

    2012-07-01

    A total of 116 samples (44 clinical specimens and 72 environmental samples) have been analyzed for the presence of Acanthamoeba. The environmental samples (ESs) were collected from four drinking water treatment plants (DWTP, n=32), seven wastewater treatment plants (n=28), and six locations of influence (n=12) on four river basins from the central area of Spain (winter-spring 2008). Water samples were concentrated by using the IDEXX Filta-Max(®) system. Acanthamoeba was identified in 65 of the 72 ESs by culture isolation (90.3%) and 63 by real-time PCR (87.5%), resulting in all sampling points (100%) positive for Acanthamoeba when considering both techniques and all the time period analyzed. Nine of the 44 clinical specimens were positive for Acanthamoeba. Seventeen Acanthamoeba strains (eight from four DWTP and nine from clinical samples) were also established in axenic-PYG medium. Twenty-four of the ESs and the 17 Acanthamoeba sp. strains were genotyped as T4/1, T4/8, and T4/9. The eight strains isolated from the DWTP samples were inoculated in nude mouse to ascertain their potential pathogenicity in this model. Animals that were inoculated died or showed central nervous system symptoms 9 days post-inoculation. Examination of immunofluorescence-stained brain and lung tissue sections showed multiple organisms invading both tissues, and re-isolation of throphozoites was successful in these tissues of all infected animals. For the first time, potentially pathogenic Acanthamoeba T4 has been detected in 100% of different types of water samples including tap water and sewage effluents in the central area of Spain suggesting a potential health threat for humans especially for the contact lens wearers.

  2. Pathogenic strains of Acanthamoeba are recognized by TLR4 and initiated inflammatory responses in the cornea.

    PubMed

    Alizadeh, Hassan; Tripathi, Trivendra; Abdi, Mahshid; Smith, Ashley Dawn

    2014-01-01

    Free-living amoebae of the Acanthamoeba species are the causative agent of Acanthamoeba keratitis (AK), a sight-threatening corneal infection that causes severe pain and a characteristic ring-shaped corneal infiltrate. Innate immune responses play an important role in resistance against AK. The aim of this study is to determine if Toll-like receptors (TLRs) on corneal epithelial cells are activated by Acanthamoeba, leading to initiation of inflammatory responses in the cornea. Human corneal epithelial (HCE) cells constitutively expressed TLR1, TLR2, TLR3, TLR4, and TLR9 mRNA, and A. castellanii upregulated TLR4 transcription. Expression of TLR1, TLR2, TLR3, and TLR9 was unchanged when HCE cells were exposed to A. castellanii. IL-8 mRNA expression was upregulated in HCE cells exposed to A. castellanii. A. castellanii and lipopolysaccharide (LPS) induced significant IL-8 production by HCE cells as measured by ELISA. The percentage of total cells positive for TLR4 was higher in A. castellanii stimulated HCE cells compared to unstimulated HCE cells. A. castellanii induced upregulation of IL-8 in TLR4 expressing human embryonic kidney (HEK)-293 cells, but not TLR3 expressing HEK-293 cells. TLR4 neutralizing antibody inhibited A. castellanii-induced IL-8 by HCE and HEK-293 cells. Clinical strains but not soil strains of Acanthamoeba activated TLR4 expression in Chinese hamster corneas in vivo and in vitro. Clinical isolates but not soil isolates of Acanthamoeba induced significant (P< 0.05) CXCL2 production in Chinese hamster corneas 3 and 7 days after infection, which coincided with increased inflammatory cells in the corneas. Results suggest that pathogenic species of Acanthamoeba activate TLR4 and induce production of CXCL2 in the Chinese hamster model of AK. TLR4 may be a potential target in the development of novel treatment strategies in Acanthamoeba and other microbial infections that activate TLR4 in corneal cells.

  3. Evaluation of the orientation of the steepest meridian of regular astigmatism among highly myopic Egyptian patients seeking non-ablative surgical correction of the refractive error

    PubMed Central

    Refai, Tamer Adel

    2015-01-01

    Introduction: LASIK surgery is currently the preferred procedure to correct low to moderate myopia. The aim of this study was to determine the orientation of the steepest meridian of regular astigmatism in order to determine the relative incidence of vertical, horizontal, and oblique regular astigmatism among highly myopic Egyptian patients seeking non-ablative surgical correction of the refractive error. Methods: One hundred and one eyes of 68 highly myopic patients who were seeking refractive surgery were included in this consecutive case series study. The refractive errors were measured using an autorefractometer and confirmed by trial. We measured the uncorrected and best corrected visual acuity in Snellen lines. Keratometry, central corneal thickness, and anterior chamber depth also were measured. The cylinder power in diopters and the axis in degrees were reported. Astigmatism was graded as with the rule (i.e., vertical meridian steeper), against the rule (i.e., horizontal meridian steeper), and oblique astigmatism. The number and the percentage of eyes with the rule, against the rule, and oblique astigmatism were calculated, and the chi-squared test was performed to analyze the data. Results: The spherical refractive error ranged from −6.5 to −24.5 diopters (−13.45 ± 4.60). The cylinder power (Cyl) ranged from −0.25 to −7.5 diopters (−2.23 ± 1.28). The uncorrected visual acuity (UCVA) in Snellen lines ranged from 0.01 – 0.1 (0.03 ± 0.02). The mean for best corrected visual acuity (BCVA) in Snellen lines was 0.40 (± 0.23). The steepest meridian was vertical (i.e., with-the-rule astigmatism) in 44 eyes (43.56%), horizontal (i.e., against-the-rule astigmatism) in 27 eyes (26.73%), and oblique (i.e., oblique astigmatism) in 30 eyes (29.70%). Conclusions: The incidence of with-the-rule astigmatism in patients with high myopia was found to be much lower than in previous studies for non-myopic patients, with a higher incidence for against

  4. Mycobacterium avium infections of Acanthamoeba strains: host strain variability, grazing-acquired infections, and altered dynamics of inactivation with monochloramine.

    PubMed

    Berry, David; Horn, Matthias; Xi, Chuanwu; Raskin, Lutgarde

    2010-10-01

    Stable Mycobacterium avium infections of several Acanthamoeba strains were characterized by increased infection resistance of recent environmental isolates and reduced infectivity in the presence of other bacteria. Exposure of M. avium in coculture with Acanthamoeba castellanii to monochloramine yielded inactivation kinetics markedly similar to those observed for A. castellanii alone.

  5. Anti-Acanthamoeba IgG, IgM, and IgA immunoreactivities in correlation to strain pathogenicity.

    PubMed

    Walochnik, J; Obwaller, A; Haller-Schober, E M; Aspöck, H

    2001-08-01

    Several representatives of the genus Acanthamoeba are known as causative agents of Acanthamoeba keratitis and granulomatous amoebic encephalitis. These occur predominantly in the immunocompromised host, but it is still unclear what primes the amoebae for pathogenicity. The aim of this study was to assess possible immunological differences between a highly pathogenic and a nonpathogenic Acanthamoeba strain. A total of 20 sera, including two sera of Acanthamoeba keratitis patients, were tested for anti-Acanthamoeba IgG, IgM, and IgA immunoreactivities using immunoblotting. All sera were positive for Acanthamoeba, revealing two predominant bands at 29 kDa and at 47 kDa, respectively. Interestingly, IgG and particularly IgA immunoreactivity enabled a clear discrimination between the pathogenic and nonpathogenic strains. Moreover, compared to the control sera, the two sera of Acanthamoeba keratitis patients showed rather weak immunoreactivities and they lacked the 29 kDa and the 47 kDa band in the IgA immunoblot against the pathogenic strain. The results of our study support the assumption that immunological predisposition might also be of importance in Acanthamoeba keratitis.

  6. Apparatus for and method of correcting for astigmatism in a light beam reflected off of a light reflecting surface

    DOEpatents

    Sawicki, Richard H.; Sweatt, William

    1987-01-01

    A technique for adjustably correcting for astigmatism in a light beam is disclosed herein. This technique utilizes first means which defines a flat, rectangular light reflecting surface having opposite reinforced side edges and which is resiliently bendable, to a limited extent, into different concave and/or convex cylindrical curvatures about a particular axis and second means acting on the first means for adjustably bending the light reflecting surface into a particular selected one of the different curvatures depending upon the astigmatism to be corrected for and for fixedly maintaining the curvature selected. In the embodiment disclosed, the light reflecting surface is adjustably bendable into the selected cylindrical curvature by application of a particular bending moment to the reinforced side edges of the light reflecting surface.

  7. Reflection and refraction of narrow Gaussian beams with general astigmatism at tilted optical surfaces: a derivation oriented toward lens design.

    PubMed

    Greco, Vincenzo; Giusfredi, Giovanni

    2007-02-01

    The formulas for the reflection and refraction of a narrow Gaussian beam with general astigmatism at a tilted optical surface are derived by ray-tracing techniques. The propagation direction of the reflected and refracted beams is computed by tracing the central ray of the incident beam, and the characteristic parameters of the respective wavefronts are worked out by applying the formulas developed for the generalized ray tracing. Moreover, the Gaussian form of the reflected and refracted amplitude distributions along the transverse coordinates is determined by requiring the matching of the incident, reflected, and refracted light spots on the optical surface. No limiting assumptions are made regarding the form of the optical interface or the orientation of the incident astigmatic wavefront. In the end, to illustrate a simple application of these formulas, the reflection of a Gaussian beam at a conicoid is considered, and a simple property of the conicoidal mirrors is reported.

  8. Management of moderate and severe corneal astigmatism with AcrySof® toric intraocular lens implantation – Our experience

    PubMed Central

    Farooqui, Javed Hussain; Koul, Archana; Dutta, Ranjan; Shroff, Noshir Minoo

    2015-01-01

    Purpose Visual performance following toric intraocular lens implantation for cataract with moderate and severe astigmatism. Setting Cataract services, Shroff Eye Centre, New Delhi, India. Design Case series. Method This prospective study included 64 eyes of 40 patients with more than 1.50 dioptre (D) of pre-existing corneal astigmatism undergoing phacoemulsification with implantation of the AcrySof® toric IntraOcular Lens (IOL). The unaided visual acuity (UCVA), best corrected visual acuity (BCVA), residual refractive sphere and refractive cylinders were evaluated. Toric IOL axis and alignment error was measured by slit lamp method and Adobe Photoshop (version 7) method. Patient satisfaction was evaluated using a satisfaction questionnaire at 3 months. Results The mean residual refractive astigmatism was 0.57 D at the final follow-up of 3 months. Mean alignment error was 3.44 degrees (SD = 2.60) by slit lamp method and 3.88 degrees (SD = 2.86) by Photoshop method. Forty-six (71.9%) eyes showed misalignment of 5 degrees or less, and 60 (93.8%) eyes showed misalignment of 10 degrees or less. The mean log MAR UCVA at 1st post-op day was 0.172 (SD = 0.02), on 7th post-op day was 0.138 (SD = 0.11), and on 30th post-op day was 0.081 (SD = 0.11). The mean log MAR BCVA at three months was −0.04 (SD = 0.76). Conclusion We believe that implantation of AcrySof® toric IOL is an effective, safe and predictable method to correct high amounts of corneal astigmatism during cataract surgery. PMID:26586976

  9. Unusual case of methicillin resistant staphylococcus aureus and acanthamoeba keratitis in a non-contact lens wearer from Kashmir, India.

    PubMed

    Lone, Rubina; Syed, Khurshid; Abdul, Rashid; Sheikh, Sajjad Ahmed; Shah, Faisal

    2009-01-01

    Acanthamoeba species can cause a chronic, progressive, ulcerative keratitis of the eye, which is not responsive to the usual antimicrobial treatment and is frequently mistaken for stromal herpes keratitis. Acanthamoeba keratitis continues to be a burgeoning and unsolved problem. Although soft contact lens wear is reported as the major risk factor in other parts of the world, reports from India suggest that acanthamoeba keratitis is more common among non-contact lens wearers. An unusual case of coinfection with Acanthamoeba and methicillin resistant staphylococcus aureus (MRSA) as causes of corneal keratitis in a contact lens wearer from Kashmir, India, is reported. Recent findings have shown that MRSA uses amoebae to spread, sidestepping hospital and other protection measures. Cysts of the isolated Acanthamoeba tolerated an incubation temperature of 40°C, indicating a pathogenic species. This case highlights the importance of culture methods in the diagnosis of corneal infection and the choice of treatment regimen.

  10. Quick survey for detection, identification and characterization of Acanthamoeba genotypes from some selected soil and water samples across Pakistan.

    PubMed

    Tanveer, Tania; Hameed, Abdul; Gul, Asma; Matin, Abdul

    2015-01-01

    Acanthamoeba is an opportunistic protozoan pathogen which is widely distributed in nature and plays a pivotal role in ecosystem. Acanthamoeba species may cause blinding keratitis and fatal granulomatous encephalitis involving central nervous system. In this study, we investigated the presence of Acanthamoeba in soil and water resources of Pakistan. Here, Acanthamoeba were recovered on non-nutrient agar plate lawn with E. coli and identified by morphological characteristics of the cyst. Furthermore PCR was performed with genus-specific primers followed by direct sequencing of the PCR product for molecular identification. Overall our PCR and sequencing results confirmed pathogenic genotypes including T4 and T15 from both soil and water samples. This is our first report of Acanthamoeba isolation from both soil and water resources of Pakistan which may serve as a potential treat to human health across the country.

  11. Detection of four adenovirus serotypes within water-isolated strains of Acanthamoeba in the Canary Islands, Spain.

    PubMed

    Lorenzo-Morales, Jacob; Coronado-Alvarez, Nieves; Martínez-Carretero, Enrique; Maciver, Sutherland K; Valladares, Basilio

    2007-10-01

    We surveyed 236 potentially pathogenic Acanthamoeba strains, isolated from water sources in the Canary Islands, for the presence of human adenoviruses (HAdV) using a polymerase chain reaction (PCR)-based typing assay. A total of 34 of these strains were found to be positive for adenovirus belonging to four different HAdV serotypes (HAdV-1, 2, 8, and 37). We found that HAdV-2 was the most frequently encountered serotype amongst the Acanthamoeba strains, and their identification was confirmed by a nested PCR specific for this serotype. We showed that Acanthamoeba genotype T4 was highly associated with serotype HAdV-2, whereas Acanthamoeba genotype T3 was most often associated with adenovirus serotypes related to ocular diseases. Based on these data, we suggest that Acanthamoeba should be considered as a potential reservoir and perhaps even a transmitter of adenoviruses to human and other secondary hosts.

  12. Isolation and Genotyping of Acanthamoeba spp. as Neglected Parasites in North of Iran

    PubMed Central

    Shokri, Azar; Sarvi, Shahabeddin; Daryani, Ahmad; Sharif, Mehdi

    2016-01-01

    Acanthamoeba, a free-living amoeba, is widely distributed in the environment, water sources, soil, dust, and air. It can cause keratitis in contact lens wearers with poor hygiene and also fatal granulomatous amebic encephalitis (GAE) in immunocompromised hosts. The aim of this study was to gain some insights into the distribution and genotypes of the potentially pathogenic species of Acanthamoeba present in water sources in north of Iran. Total 43 Acanthamoeba species were isolated from 77 water samples taken from different water sources within the Mazandaran province in Northern Iran (Sari city and suburbs). Isolates were identified based on cyst and trophozoite morphological characteristics as well genetics. PCR fragments corresponding to the small-subunit 18S rRNA gene were sequenced for 20 of 43 positive isolates. The results revealed that 83.3% of sequenced isolates belonged to the T4 genotype and the rest belonged to the T2 genotype. Our results indicated that Acanthamoeba is widely distributed in Sari city. As the incidence in Iran of amoebic keratitis has increased in recent years, the exact estimation of the prevalence of this amoeba and its predominant genotype may play a crucial role in prevention of the disease. Sari city has several rivers, seashores, and natural recreational amenities, which attract visitors during the year. This is the first report of Acanthamoeba genotypes from water sources in Sari city, Mazandaran province of Iran, and the results suggest that more attention is needed to protect the visiting population and immunocompromised individuals. PMID:27658596

  13. Riboflavin and ultraviolet-A as adjuvant treatment against Acanthamoeba cysts

    PubMed Central

    Lamy, Ricardo; Chan, Elliot; Good, Samuel D; Cevallos, Vicky; Porco, Travis C; Stewart, Jay M

    2015-01-01

    Background Experimental studies have shown that the standard dose of R or R+UVA as solo treatment are not able to exterminate Acanthamoeba cysts or even trophozoites. The purpose of this study is to determine whether the application of R+UVA can enhance the cysticidal effects of cationic antiseptic agents in vitro. Methods The log of either polyhexamethylene biguanide (PHMB) or chlorhexidine minimal cysticidal concentration (MCC) in solutions containing riboflavin (concentrations 0.1 %; 0.05% and 0.025 %) plus either Acanthamoeba castellanii cysts or Acanthamoeba polyphaga cysts was determined and compared in groups treated with UVA 30 mW/cm2 for 30 min and in control groups (with no exposure to UVA). A permutation test was used to determine the P-value associated with treatment. Results Regardless of the riboflavin concentration and UVA treatment condition, no trophozoites were seen in plates where the cysts were previously exposed to cationic antiseptic agents concentrations ≥ 200 µg/mL for Acanthamoeba castellanii samples and ≥ 100 µg/mL for Acanthamoeba polyphaga samples. There was no statistical evidence that R+UVA treatment was associated with MCC (P = 0.82). Conclusion R+UVA in doses up to 10 times higher than recommended for corneal crosslinking does not enhance the cysticidal effect of either polyhexamethylene biguanide or chlorhexidine in vitro. PMID:26355273

  14. Amoebicidal activity of caffeine and maslinic acid by the induction of Programmed Cell Death in Acanthamoeba.

    PubMed

    Martín-Navarro, Carmen M; López-Arencibia, Atteneri; Sifaoui, Ines; Reyes-Battle, María; Fouque, Emilie; Osuna, Antonio; Valladares, Basilio; Piñero, José E; Héchard, Yann; Maciver, Sutherland K; Lorenzo-Morales, Jacob

    2017-03-20

    Free living amoebae of the genus Acanthamoeba are the causal agents of a sight threatening ulceration of the cornea called Acanthamoeba keratitis, and the rare but usually fatal granulomatous amoebic encephalitis. Although there are many therapeutic options for the treatment of Acanthamoeba infections, they are generally lengthy and/or have limited efficacy. For the best clinical outcome, the treatments should target both the trophozoite and the cyst stages as the later are known to confer resistance to treatment. In this study we document the activity of caffeine and maslinic acid against both the trophozoite and the cyst stages of three clinical strains of Acanthamoeba These drugs were chosen because they are reported to inhibit glycogen phosphorylase which is required for encystation. Maslinic acid is also reported to be an inhibitor of extracellular proteases which may be relevant since the protease activity of Acanthamoeba is correlated with their pathogenicity. We also provide evidence or the first time that both drugs exert their anti-amoebal effects through programmed cell death.

  15. Isolation and genotyping of acanthamoeba strains from soil sources from Jamaica, West Indies.

    PubMed

    Todd, Cheridah D; Reyes-Batlle, María; Martín-Navarro, Carmen Ma; Dorta-Gorrín, Alexis; López-Arencibia, Atteneri; Martínez-Carretero, Enrique; Piñero, José E; Valladares, Basilio; Lindo, John F; Lorenzo-Morales, Jacob

    2015-01-01

    Acanthamoeba spp. are opportunistic pathogens that are ubiquitous in nature. Many species of this genus are responsible for a fatal encephalitis and keratitis in humans and other animals. Seventy-two soil samples were collected from the parishes across Jamaica and assessed for the presence of Acanthamoeba spp. Cultivation was carried out on non-nutrient agar plates seeded with heat killed Escherichia coli. PCR and sequencing of the DF3 region were carried out in order to genotype the isolated strains of Acanthamoeba. Thermotolerance and osmotolerance assays were utilized to investigate the pathogenic potential of the Acanthamoeba isolates. Acanthamoeba spp. was isolated from 63.9% of soil samples. Sequencing of the DF3 region of the 18S rDNA resulted in the identification of genotypes T4, T5, and T11. T4 genotype was most frequently isolated. Most isolates were thermotolerant or both thermotolerant and osmotolerant, indicating that they may present the potential to cause disease in humans and other animals.

  16. Identification and typing of free-living Acanthamoeba spp. by MALDI-TOF MS Biotyper.

    PubMed

    Del Chierico, Federica; Di Cave, David; Accardi, Cristel; Santoro, Maristella; Masotti, Andrea; D'Alfonso, Rossella; Berrilli, Federica; Urbani, Andrea; Putignani, Lorenza

    2016-11-01

    Over the years, the potential pathogenicity of Acanthamoeba for humans and animals has gained increasing attention from the scientific community. More than 24 species belong to this genus, however only some of them are causative agents of keratitis and encephalitis in humans. Due to technical difficulties in diagnosis, these infections are likely to be under-detected. The introduction of 18S rDNA amplification for the identification of Acanthamoeba has dramatically enhanced diagnosis performances, but the attestation of genotyping requires supplementary sequencing-based procedures. In this study, 15 Acanthamoeba strains were collected and grown on nutrient agar media. Each strain was genotyped by end-point PCR assay for the amplification of the 18S rDNA gene and the genotype was assigned by sequencing analysis through neighbor joining phylogenetic tree. In order to optimize standardization of the MALDI-TOF MS assay, we established the collection time point at the cystic phase. Two strains of each genotype were randomly chosen to customize the biotyper database. For all strains, 24 spectral measurements were acquired and submitted to identification and cluster analysis of spectra. The obtained results highlighted the correct identification of Acanthamoeba strains and the overlapping of spectra dendrogram clusters to the 18S genotype assignations. In conclusion, the MALDI-TOF MS Biotyper revealed the capability to identify and genotype the Acanthamoeba strains, providing a new frontier in the diagnostic identification of amaebae and in taxonomic and phylogenetic studies.

  17. A multisystemic Acanthamoeba infection in a dog in Tenerife, Canary Islands, Spain.

    PubMed

    Valladares, María; Reyes-Batlle, María; Mora-Peces, Inmaculada; Martín-Navarro, Carmen M; López-Arencibia, Atteneri; Dorta-Gorrín, Alexis; Comyn-Afonso, Estefanía; Martínez-Carretero, Enrique; Maciver, Sutherland K; Piñero, José E; Valladares, Basilio; Lorenzo-Morales, Jacob

    2014-10-15

    A 22-month-old male Spanish water dog was hospitalized after its physical examination revealed fever and movement difficulty. After 24h, the dog was found to have a high fever (39.5 °C) and was treated empirically with doxycycline/ciprofloxacin. At 48 h, after submission the fever rose to 41 °C and the animal presented with a stiff neck and dehydration. Peripheral blood and cerebrospinal fluid (CSF) were sampled and trophozoites with an Acanthamoeba-like morphology were observed in the CSF. PCR specific for Acanthamoeba, Naegleria fowleri and Balamuthia mandrillaris were performed and the CSF sample found positive for Acanthamoeba. Lungs, kidney, liver and spleen samples were collected post mortem. All collected organ samples were positive for Acanthamoeba by PCR, thus confirming a multisystemic infection. Water samples taken at a suspected site of infection yielded an almost identical PCR fragment to those of the clinical samples, indicating that this was probably where the infection originated. This is the first report of a fatal case of Acanthamoeba disseminated infection in a dog in Spain.

  18. Physiological, morphological, and immunochemical parameters used for the characterization of clinical and environmental isolates of Acanthamoeba.

    PubMed

    Becker-Finco, A; Costa, A O; Silva, S K; Ramada, J S; Furst, C; Stinghen, A E; De Figueiredo, B C; De Moura, J; Alvarenga, L M

    2013-03-01

    The factors that characterize Acanthamoeba strains as harmless or potentially pathogenic have not been elucidated. Analysing the in vitro and in vivo parameters of Acanthamoeba samples, including heat tolerance at temperatures close to that of the human body, cytopathic effects, and their ability to cause infections in animals, has been proposed to identify their pathogenic potential. Another promising criterion for differentiating strains is the analysis of their biochemical and immunochemical properties. In this study, a comparative evaluation between clinical and environmental Acanthamoeba isolates was performed on the basis of physiological, morphological, and immunochemical criteria. Crude antigens were used to characterize the protein profiles by electrophoresis and immunize mice to produce polyclonal and monoclonal antibodies. The antibodies were characterized using ELISA, Western blotting, and immunofluorescence techniques. The results obtained with polyclonal antibodies suggest the presence of specific proteins for each studied isolate and co-reactive immunochemical profiles among conserved components. Ten monoclonal antibody clones were obtained; mAb3 recognized 3 out of 4 samples studied. The results of this study may help standardize criteria for identifying and characterizing Acanthamoeba strains. Taken together, our results support the view that a set of features may help differentiate Acanthamoeba species and isolates.

  19. Isolation and Genotyping of Acanthamoeba spp. as Neglected Parasites in North of Iran.

    PubMed

    Shokri, Azar; Sarvi, Shahabeddin; Daryani, Ahmad; Sharif, Mehdi

    2016-08-01

    Acanthamoeba, a free-living amoeba, is widely distributed in the environment, water sources, soil, dust, and air. It can cause keratitis in contact lens wearers with poor hygiene and also fatal granulomatous amebic encephalitis (GAE) in immunocompromised hosts. The aim of this study was to gain some insights into the distribution and genotypes of the potentially pathogenic species of Acanthamoeba present in water sources in north of Iran. Total 43 Acanthamoeba species were isolated from 77 water samples taken from different water sources within the Mazandaran province in Northern Iran (Sari city and suburbs). Isolates were identified based on cyst and trophozoite morphological characteristics as well genetics. PCR fragments corresponding to the small-subunit 18S rRNA gene were sequenced for 20 of 43 positive isolates. The results revealed that 83.3% of sequenced isolates belonged to the T4 genotype and the rest belonged to the T2 genotype. Our results indicated that Acanthamoeba is widely distributed in Sari city. As the incidence in Iran of amoebic keratitis has increased in recent years, the exact estimation of the prevalence of this amoeba and its predominant genotype may play a crucial role in prevention of the disease. Sari city has several rivers, seashores, and natural recreational amenities, which attract visitors during the year. This is the first report of Acanthamoeba genotypes from water sources in Sari city, Mazandaran province of Iran, and the results suggest that more attention is needed to protect the visiting population and immunocompromised individuals.

  20. Acanthamoeba T4 and T15 genotypes associated with keratitis infections in Italy.

    PubMed

    Di Cave, D; Monno, R; Bottalico, P; Guerriero, S; D'Amelio, S; D'Orazi, C; Berrilli, F

    2009-06-01

    Thus far there is little data available concerning Acanthamoeba associated amoebic keratitis (AK) from Italy. In order to understand the incidence of Acanthamoeba in patients with ocular infections and to characterize the isolates at the molecular level, ocular specimens and contact lenses or lens case solutions from 140 patients were analysed by culture and by an 18S rRNA (Rns) gene-based PCR method. Nineteen (13.6%) patients showed Acanthamoeba culture positive samples. Eleven out of the 14 genetically characterized isolates were assigned to the T4 genotype. Three isolates, two of them from patients with keratitis responding to specific anti-Acanthamoeba therapy, were identified as belonging to the T15 genotype. This finding represents the first association between the T15 genotype and human amoebic keratitis. PCR amplification of the 18S ribosomal DNA proved to be a sensitive method, potentially able to detect Acanthamoeba without the need of long culture incubation, and thus considerably useful for clinical applications.

  1. Pathogenic and opportunistic free-living amoebae: Acanthamoeba spp., Balamuthia mandrillaris, Naegleria fowleri, and Sappinia diploidea.

    PubMed

    Visvesvara, Govinda S; Moura, Hercules; Schuster, Frederick L

    2007-06-01

    Among the many genera of free-living amoebae that exist in nature, members of only four genera have an association with human disease: Acanthamoeba spp., Balamuthia mandrillaris, Naegleria fowleri and Sappinia diploidea. Acanthamoeba spp. and B. mandrillaris are opportunistic pathogens causing infections of the central nervous system, lungs, sinuses and skin, mostly in immunocompromised humans. Balamuthia is also associated with disease in immunocompetent children, and Acanthamoeba spp. cause a sight-threatening infection, Acanthamoeba keratitis, mostly in contact-lens wearers. Of more than 30 species of Naegleria, only one species, N. fowleri, causes an acute and fulminating meningoencephalitis in immunocompetent children and young adults. In addition to human infections, Acanthamoeba, Balamuthia and Naegleria can cause central nervous system infections in animals. Because only one human case of encephalitis caused by Sappinia diploidea is known, generalizations about the organism as an agent of disease are premature. In this review we summarize what is known of these free-living amoebae, focusing on their biology, ecology, types of disease and diagnostic methods. We also discuss the clinical profiles, mechanisms of pathogenesis, pathophysiology, immunology, antimicrobial sensitivity and molecular characteristics of these amoebae.

  2. Cysteine protease inhibitor (AcStefin) is required for complete cyst formation of Acanthamoeba.

    PubMed

    Lee, Jung-Yub; Song, Su-Min; Moon, Eun-Kyung; Lee, Yu-Ran; Jha, Bijay Kumar; Danne, Dinzouna-Boutamba Sylvatrie; Cha, Hee-Jae; Yu, Hak Sun; Kong, Hyun-Hee; Chung, Dong-Il; Hong, Yeonchul

    2013-04-01

    The encystation of Acanthamoeba leads to the formation of resilient cysts from vegetative trophozoites. This process is essential for parasite survival under unfavorable conditions, such as those associated with starvation, low temperatures, and biocides. Furthermore, cysteine proteases have been implicated in the massive turnover of intracellular components required for encystation. Thus, strict modulation of the activities of cysteine proteases is required to protect Acanthamoeba from intracellular damage. However, mechanisms underlying the control of protease activity during encystation have not been established in Acanthamoeba. In the present study, we identified and characterized Acanthamoeba cysteine protease inhibitor (AcStefin), which was found to be highly expressed during encystation and to be associated with lysosomes by fluorescence microscopy. Recombinant AcStefin inhibited various cysteine proteases, including human cathepsin B, human cathepsin L, and papain. Transfection with small interfering RNA against AcStefin increased cysteine protease activity during encystation and resulted in incomplete cyst formation, reduced excystation efficiency, and a significant reduction in cytoplasmic area. Taken together, these results indicate that AcStefin is involved in the modulation of cysteine proteases and that it plays an essential role during the encystation of Acanthamoeba.

  3. Failure of chemotherapy in the first reported cases of Acanthamoeba keratitis in Pakistan.

    PubMed

    Siddiqui, Ruqaiyyah; Chaudhry, Tanveer; Lakhundi, Sahreena; Ahmad, Khabir; Khan, Naveed Ahmed

    2014-01-01

    Acanthamoeba keratitis is a painful and progressive infection of the cornea that can result in loss of vision. Here, for the first time in Pakistan, we report two cases of Acanthamoeba keratitis. The first patient was a 37-year-old female who presented with severe itching, redness, pain, along with loss of vision. The patient was a regular soft contact lens wearer. The second patient was a 25-year-old female who had been using soft contact lenses for the past two years. She presented with a burning sensation and extreme pain, along with loss of vision. Both patients were treated for a possible microbial keratitis with topical moxifloxacin hydrochloride drops, vancomycin drops, propamidine isethionate ointment, amphotericin B drops, and amikacin drops. However, the response was inadequate and both patients were referred for corneal transplant. Acanthamoeba castellanii was isolated by placing contact lenses and contact lens cases on non-nutrient agar plates containing a lawn of non-invasive Escherichia coli K-12 HB101 bacteria. The polymerase chain reaction (PCR) using genus-specific probes confirmed the identity of Acanthamoeba spp., whereas the morphological characteristics of trophozoites and cysts were suggestive of A. castellanii in both cases. With growing use of contact lenses for vision correction/cosmetic use coupled with sub-standard lens care in this region and the possibility of non-contact lens-associated Acanthamoeba keratitis, a need for increased awareness of this sight-threatening infection is discussed further.

  4. Cysteine Protease Inhibitor (AcStefin) Is Required for Complete Cyst Formation of Acanthamoeba

    PubMed Central

    Lee, Jung-Yub; Song, Su-Min; Moon, Eun-Kyung; Lee, Yu-Ran; Jha, Bijay Kumar; Danne, Dinzouna-Boutamba Sylvatrie; Cha, Hee-Jae; Yu, Hak Sun; Kong, Hyun-Hee; Chung, Dong-Il

    2013-01-01

    The encystation of Acanthamoeba leads to the formation of resilient cysts from vegetative trophozoites. This process is essential for parasite survival under unfavorable conditions, such as those associated with starvation, low temperatures, and biocides. Furthermore, cysteine proteases have been implicated in the massive turnover of intracellular components required for encystation. Thus, strict modulation of the activities of cysteine proteases is required to protect Acanthamoeba from intracellular damage. However, mechanisms underlying the control of protease activity during encystation have not been established in Acanthamoeba. In the present study, we identified and characterized Acanthamoeba cysteine protease inhibitor (AcStefin), which was found to be highly expressed during encystation and to be associated with lysosomes by fluorescence microscopy. Recombinant AcStefin inhibited various cysteine proteases, including human cathepsin B, human cathepsin L, and papain. Transfection with small interfering RNA against AcStefin increased cysteine protease activity during encystation and resulted in incomplete cyst formation, reduced excystation efficiency, and a significant reduction in cytoplasmic area. Taken together, these results indicate that AcStefin is involved in the modulation of cysteine proteases and that it plays an essential role during the encystation of Acanthamoeba. PMID:23397569

  5. Chloroquine Has a Cytotoxic Effect on Acanthamoeba Encystation through Modulation of Autophagy

    PubMed Central

    Jha, Bijay Kumar; Jung, Hui-Jung; Seo, Incheol; Kim, Hyun Ah; Suh, Seong-Il; Suh, Min-Ho

    2014-01-01

    Encystation of Acanthamoeba castellanii is associated with resistance to chemotherapeutic agents. Blocking the encystation process could potentiate the efficacy of chemotherapeutic agents and biocides. During encystation, autophagy is highly stimulated and required for proper encystation of Acanthamoeba. In this study, the cytotoxic effect of chloroquine, a well-known autophagy-inhibitory drug, was tested in A. castellanii. Chloroquine was able to selectively reduce cell survival during the encystation of A. castellanii. However, A. castellanii trophozoites and mature cysts were resistant to chloroquine. Chloroquine treatment led to an increase in the number and size of lysosomes in encysting cells. Moreover, chloroquine inhibited the degradation of long-lived proteins in the encysting cells. Decreased autophagic flux, indicated by an increased number of lysosomes and decreased degradation of long-lived proteins, may be the mechanism by which cell death is induced by chloroquine in encysting Acanthamoeba. These results suggest a potential novel therapeutic application of chloroquine as an anti-Acanthamoeba drug. Our findings also suggest that targeting autophagy could be a therapeutic strategy against Acanthamoeba infection. PMID:25114131

  6. Chloroquine has a cytotoxic effect on Acanthamoeba encystation through modulation of autophagy.

    PubMed

    Jha, Bijay Kumar; Jung, Hui-Jung; Seo, Incheol; Kim, Hyun Ah; Suh, Seong-Il; Suh, Min-Ho; Baek, Won-Ki

    2014-10-01

    Encystation of Acanthamoeba castellanii is associated with resistance to chemotherapeutic agents. Blocking the encystation process could potentiate the efficacy of chemotherapeutic agents and biocides. During encystation, autophagy is highly stimulated and required for proper encystation of Acanthamoeba. In this study, the cytotoxic effect of chloroquine, a well-known autophagy-inhibitory drug, was tested in A. castellanii. Chloroquine was able to selectively reduce cell survival during the encystation of A. castellanii. However, A. castellanii trophozoites and mature cysts were resistant to chloroquine. Chloroquine treatment led to an increase in the number and size of lysosomes in encysting cells. Moreover, chloroquine inhibited the degradation of long-lived proteins in the encysting cells. Decreased autophagic flux, indicated by an increased number of lysosomes and decreased degradation of long-lived proteins, may be the mechanism by which cell death is induced by chloroquine in encysting Acanthamoeba. These results suggest a potential novel therapeutic application of chloroquine as an anti-Acanthamoeba drug. Our findings also suggest that targeting autophagy could be a therapeutic strategy against Acanthamoeba infection.

  7. Role of carbohydrate-mediated adherence in cytopathogenic mechanisms of Acanthamoeba.

    PubMed

    Cao, Z; Jefferson, D M; Panjwani, N

    1998-06-19

    Acanthamoeba keratitis is a vision-threatening corneal infection. The mannose-binding protein of Acanthamoeba is thought to mediate adhesion of parasites to host cells. We characterized the amoeba lectin with respect to its carbohydrate binding properties and the role in amoeba-induced cytopathic effect (CPE). Sugar inhibition assays revealed that the amoeba lectin has the highest affinity for alpha-Man and Man(alpha1-3)Man units. In vitro cytopathic assays indicated that mannose-based saccharides which inhibit amoeba adhesion to corneal epithelial cells were also potent inhibitors of amoeba-induced CPE. Another major finding was that N-acetyl-D-glucosamine (GlcNAc) which does not inhibit adhesion of amoeba to host cells is also an inhibitor of amoeba-induced CPE. The Acanthamoebae are thought to produce CPE by secreting cytotoxic proteinases. By zymography, one metalloproteinase and three serine proteinases were detected in the conditioned media obtained after incubating amoebae with the host cells. The addition of free alpha-Man and GlcNAc to the co-culture media inhibited the secretion of the metalloproteinase and serine proteinases, respectively. In summary, we have shown that the lectin-mediated adhesion of the Acanthamoeba to host cells is a prerequisite for the amoeba-induced cytolysis of target cells and have implicated a contact-dependent metalloproteinase in the cytopathogenic mechanisms of Acanthamoeba.

  8. Evaluation of the effectiveness of tea tree oil in treatment of Acanthamoeba infection.

    PubMed

    Hadaś, Edward; Derda, Monika; Cholewiński, Marcin

    2017-03-01

    Eye diseases caused by amoebae from the genus Acanthamoeba are usually chronic and severe, and their treatment is prolonged and not very effective. The difficulties associated with therapy have led to attempts at finding alternative treatment methods. Particularly popular is searching for cures among drugs made of plants. However, no substances with total efficacy in treating Acanthamoeba keratitis have been identified.Results of our semi in vivo studies of tea tree oil simulating eyeball infection demonstrated 100% effectiveness in the case of both trophozoites and cysts of amoebae from the genus Acanthamoeba. The action of tea tree oil indicates that this is the first substance with a potential ability to quickly and effectively remove the amoebae from the eye. Tea tree oil has the ability to penetrate tissues, which allows it to destroy amoebae in both the shallow and deep layers of the cornea. The present research into the use of tea tree oil in the therapy of Acanthamoeba infection is the first study of this type in parasitology. It offers tremendous potential for effective treatment of Acanthamoeba keratitis and other diseases caused by these protozoa.

  9. Acanthamoeba genotypes T3 and T4 as causative agents of amoebic keratitis in Mexico.

    PubMed

    Omaña-Molina, Maritza; Vanzzini-Zago, Virginia; Hernandez-Martinez, Dolores; Gonzalez-Robles, Arturo; Salazar-Villatoro, Lizbeth; Ramirez-Flores, Elizabeth; Oregon-Miranda, Eric; Lorenzo-Morales, Jacob; Martinez-Palomo, Adolfo

    2016-02-01

    Free-living amoebae (FLA) are widely distributed worldwide. Some genera included in this group act as opportunistic pathogens causing fatal encephalitis and Acanthamoeba keratitis (AK), a sight-threatening infection of the cornea associated with the use of soft contact lenses that could even end in blindness if an early diagnosis and treatment are not achieved. Furthermore, the numbers of AK cases keep rising worldwide mainly due to an increase of contact lens wearers and lack of hygiene in the maintenance of lenses and their cases. In Mexico, no cases of AK have been described so far although the isolation of other pathogenic FLA such as Naegleria fowleri and Balamuthia mandrillaris from both clinical and environmental sources has been reported. The present study reports two cases of Acanthamoeba keratitis diagnosed in two patients admitted to the Hospital "Luis Sánchez Bulnes" for Blindness Prevention in Mexico City, Mexico. Corneal scrapes and contact lenses were checked for the presence of Acanthamoeba strains in both patients. Strains were axenized after initial isolation to classify at the genotype level. After sequencing the diagnostic fragment 3 (DF3) region located on the 18S ribosomal DNA (rDNA) gene of Acanthamoeba, genotype T3 and genotype T4 were identified in clinical case 1 and 2, respectively. To our knowledge, these are the first reported cases of AK in Mexico in the literature and the first description of Acanthamoeba genotypes T3 and T4 as causative agents of amoebic infection.

  10. Detection and quantification of human adenovirus genomes in Acanthamoeba isolated from swimming pools.

    PubMed

    Staggemeier, Rodrigo; Arantes, Thalita; Caumo, Karin S; Rott, Marilise B; Spilki, Fernando R

    2016-01-01

    Acanthamoeba is the most common free-living environmental amoeba, it may serve as an important vehicle for various microorganisms living in the same environment, such as viruses, being pathogenic to humans. This study aimed to detect and quantify human adenoviruses (HAdV) in Acanthamoebas isolated from water samples collected from swimming pools in the city of Porto Alegre, Southern Brazil. Free-living amoebae of the genus Acanthamoeba were isolated from water samples, and isolates (n=16) were used to investigate the occurrence of HAdVs. HAdV detection was performed by quantitative real-time polymerase chain reaction (qPCR). HAdVs were detected in 62.5% (10/16) of Acanthamoeba isolates, ranging from 3.24x103 to 5.14x105 DNA copies per milliliter of isolate. HAdV viral loads found in this study are not negligible, especially because HAdV infections are associated with several human diseases, including gastroenteritis, respiratory distress, and ocular diseases. These findings reinforce the concept that Acanthamoeba may act as a reservoir and promote HAdV transmission through water.

  11. Combination of Toric and multifocal intraocular lens implantation in bilateral cataract patients with unilateral astigmatism

    PubMed Central

    Liang, Jing-Li; Tian, Fang; Zhang, Hong; Teng, He

    2016-01-01

    AIM To assess the binocular visual function in bilateral cataract patients with unilateral astigmatism after combined implantations of Toric with multifocal intraocular lens (IOL), and to compare with that of Toric and monofocal IOL implantation. METHODS All the 30 patients with unilateral astigmatism suffered bilateral cataract were randomly divided into two groups: Toric plus multifocal IOL group and Toric plus monofocal IOL group. Uncorrected and corrected visual acuity at distance (5.0 m), intermediate distance (0.6 m), and near (0.33 m), contrast sensitivity, and stereopsis were assessed 6mo after surgery. Patients were also surveyed for visual disturbances and spectacle dependence. RESULTS Binocular uncorrected visual acuity (LogMAR) of Toric/multifocal IOL eyes at distance, intermediate, near were 0.05±0.05, 0.24±0.10, and 0.14±0.06 respectively. The values of Toric plus monofocal IOL eyes were 0.06±0.07, 0.26±0.08, and 0.37±0.10 respectively. These values did not indicate significant differences between two groups with exception of near visual acuity. In the photopic condition (with or without glare), the contrast sensitivity of multifocal IOL eyes was significant lower than the monofocal IOL eyes in 18 cpd. In the mesopic condition, the contrast sensitivity of multifocal group was significant lower than monofocal group in 12 cpd, and in mesopic glare condition, this significant difference was found both in 6 cpd and 12 cpd. The stereopsis of Toric/multifocal IOL eyes decreased slightly (100±80 seconds of arc, t=2.222, P=0.136). Mean near vision for patient satisfaction was statistically significantly higher in Toric/multifocal IOL group patients versus than that in Toric/monofocal IOL group (80% vs 25.5%, P=0.000). Visual disturbance was not noticed in either group. CONCLUSION Although the combination of Toric and multifocal IOL implantation results in compromising stereoacuity, it can still provide patients with high levels of spectacle freedom and

  12. The level of improvement of visual acuity in high corneal astigmatism with rigid gas permeable contact lenses.

    PubMed

    Opačić, Dalibor; Miljak, Snježana; Ćuruvija-Opačić, Ksenija

    2015-03-01

    The aim of this study was to calculate the level of improvement of visual acuity comparing the best corrected visual acuity (VA) achieved with spectacles with the best corrected VA achieved with rigid gas permeable (RGP) contact lenses in patients with high, simple or compound corneal astigmatism (myopic, hypermetropic and mixed). The investigation of patients included auto-kerato-refractometry, manual keratometry, corneal topography and visual acuity with Snellen chart. The best corrected VA obtained with spectacles was compared with the best corrected VA obtained with RGP contact lenses in 72 patients (116 eyes). All patients showed a significant improvement in visual acuity with RGP lenses from one to seven lines compared to spectacles (p = 0.0001). Level of improvement in VA represented as the number of lines obtained was as follows: 74 percent of patients got two to four lines more in VA with RGP lenses compared to spectacles, and almost 10 percent of patients got five to seven lines. RGP contact lenses provide a significant improvement in VA compared to VA reached with spectacles in patients with high corneal astigmatism. The benefit in VA with RGP lenses is higher as the astigmatism is higher.

  13. Advanced astigmatism-corrected tandem Wadsworth mounting for small-scale spectral broadband imaging spectrometer.

    PubMed

    Lei, Yu; Lin, Guan-yu

    2013-01-01

    Tandem gratings of double-dispersion mount make it possible to design an imaging spectrometer for the weak light observation with high spatial resolution, high spectral resolution, and high optical transmission efficiency. The traditional tandem Wadsworth mounting is originally designed to match the coaxial telescope and large-scale imaging spectrometer. When it is used to connect the off-axis telescope such as off-axis parabolic mirror, it presents lower imaging quality than to connect the coaxial telescope. It may also introduce interference among the detector and the optical elements as it is applied to the short focal length and small-scale spectrometer in a close volume by satellite. An advanced tandem Wadsworth mounting has been investigated to deal with the situation. The Wadsworth astigmatism-corrected mounting condition for which is expressed as the distance between the second concave grating and the imaging plane is calculated. Then the optimum arrangement for the first plane grating and the second concave grating, which make the anterior Wadsworth condition fulfilling each wavelength, is analyzed by the geometric and first order differential calculation. These two arrangements comprise the advanced Wadsworth mounting condition. The spectral resolution has also been calculated by these conditions. An example designed by the optimum theory proves that the advanced tandem Wadsworth mounting performs excellently in spectral broadband.

  14. Non-astigmatic imaging with matched pairs of spherically bent reflectors

    DOEpatents

    Bitter, Manfred Ludwig [Princeton, NJ; Hill, Kenneth Wayne [Plainsboro, NJ; Scott, Steven Douglas [Wellesley, MA; Feder, Russell [Newton, PA; Ko, Jinseok [Cambridge, MA; Rice, John E [N. Billerica, MA; Ince-Cushman, Alexander Charles [New York, NY; Jones, Frank [Manalapan, NJ

    2012-07-10

    Arrangements for the point-to-point imaging of a broad spectrum of electromagnetic radiation and ultrasound at large angles of incidence employ matched pairs of spherically bent reflectors to eliminate astigmatic imaging errors. Matched pairs of spherically bent crystals or spherically bent multi-layers are used for X-rays and EUV radiation; and matched pairs of spherically bent mirrors that are appropriate for the type of radiation are used with microwaves, infrared and visible light, or ultrasound. The arrangements encompass the two cases, where the Bragg angle--the complement to the angle of incidence in optics--is between 45.degree. and 90.degree. on both crystals/mirrors or between 0.degree. and 45.degree. on the first crystal/mirror and between 45.degree. and 90.degree. on the second crystal/mirror, where the angles of convergence and divergence are equal. For x-rays and EUV radiation, also the Bragg condition is satisfied on both spherically bent crystals/multi-layers.

  15. Macroscopic three-dimensional particle location using stereoscopic imaging and astigmatic aberrations.

    PubMed

    Fuchs, Thomas; Hain, Rainer; Kähler, Christian J

    2014-12-15

    This Letter presents a stereoscopic imaging concept for measuring the locations of particles in three-dimensional space. The method is derived from astigmatism particle tracking velocimetry (APTV), a powerful technique that is capable of determining 3D particle locations with a single camera. APTV locates particle xy coordinates with high accuracy, while the particle z coordinate has a larger location uncertainty. This is not a problem for 3D2C (i.e., three dimensions, two velocity components) measurements, but for highly three-dimensional flows, it is desirable to measure three velocity components with similar accuracy. The stereoscopic APTV approach discussed in this report has this capability. The technique employs APTV for giving an initial estimate of the particle locations. With this information, corresponding particle images on both sensors of the stereoscopic imaging system are matched. Particle locations are then determined by mapping the two particle image sensor locations to physical space. The measurement error of stereo APTV, determined by acquiring images of 1-μm DEHS particles in a 40 mm×40 mm×20 mm measurement volume in air at Δxyz→0 between two frames, is less than 0.012 mm for xy and 0.025 mm for z. This error analysis proves the excellent suitability of stereo APTV for the measurement of three-dimensional flows in macroscopic domains.

  16. Inspection focus technology of space tridimensional mapping camera based on astigmatic method

    NASA Astrophysics Data System (ADS)

    Wang, Zhi; Zhang, Liping

    2010-10-01

    The CCD plane of the space tridimensional mapping camera will be deviated from the focal plane(including the CCD plane deviated due to camera focal length changed), under the condition of space environment and vibration, impact when satellite is launching, image resolution ratio will be descended because defocusing. For tridimensional mapping camera, principal point position and focal length variation of the camera affect positioning accuracy of ground target, conventional solution is under the condition of vacuum and focusing range, calibrate the position of CCD plane with code of photoelectric encoder, when the camera defocusing in orbit, the magnitude and direction of defocusing amount are obtained by photoelectric encoder, then the focusing mechanism driven by step motor to compensate defocusing amount of the CCD plane. For tridimensional mapping camera, under the condition of space environment and vibration, impact when satellite is launching, if the camera focal length changes, above focusing method has been meaningless. Thus, the measuring and focusing method was put forward based on astigmation, a quadrant detector was adopted to measure the astigmation caused by the deviation of the CCD plane, refer to calibrated relation between the CCD plane poison and the asrigmation, the deviation vector of the CCD plane can be obtained. This method includes all factors caused deviation of the CCD plane, experimental results show that the focusing resolution of mapping camera focusing mechanism based on astigmatic method can reach 0.25 μm.

  17. Lethal Effects of Helianthemum lippii (L.) on Acanthamoeba castellanii Cysts in Vitro

    PubMed Central

    Badria, F.A.; Hetta, M.H.; Sarhan, Rania M.; Ezz El-Din, M.H.

    2014-01-01

    Acanthamoeba spp. commonly cause Acanthamoeba keratitis which is typically associated with the wear of contact lenses. Therefore, finding an economic, efficient, and safe therapy of natural origin is of outmost importance. This study examined the in vitro lethal potential of ethyl acetate and methanol extracts of Helianthemum lippii (L.) (sun roses) against Acanthamoeba castellanii cysts isolated from patients with amoebic keratitis. Both extracts proved to be potent as regard to their lethal effects on A. castellanii cysts with comparable results to chlorhexidine. The ethyl acetate was more promising with cumulative lethality. It showed a highly significant lethal percentage along the duration of treatment. The analysis of the more potent ethyl acetate extract revealed the presence of 2.96 mg/100 g of total phenolics, 0.289 mg/100 ml of total flavonoids and 37 mg/100 mg of total tannins which highlighted their phytomedicinal role. PMID:25031463

  18. Isolation and identification of Acanthamoeba species from natural water sources in the northeastern part of Thailand.

    PubMed

    Thammaratana, Thani; Laummaunwai, Porntip; Boonmars, Thidarut

    2016-04-01

    Acanthamoeba are found in the environment, particularly in water, all over the world. The genus is currently classified into 20 different genotypes, T1-T20. In this study, 63 natural water samples from 11 provinces in northeast Thailand were collected and cultured on non-nutrient agar plates. Positive samples by culture were subsequently analyzed by molecular methods. The identification of Acanthamoeba was based on morphological features and molecular techniques using PCR and DNA sequencing. The results showed that 10 samples out of 63 were positive (15.9 %). Phylogenetic analysis revealed that seven samples were T4, one sample was similar to T3, and the other two samples were similar to T5. This is the first report demonstrating the contamination of Acanthamoeba species in natural water sources in northeast Thailand.

  19. 18S ribosomal DNA genotypes of Acanthamoeba species isolated from contact lens cases in the Philippines.

    PubMed

    Rivera, Windell L; Adao, Davin Edric V

    2009-10-01

    This study was carried out to document the genotypes of Acanthamoeba present in contact lens cases from 50 randomly selected contact lens wearers living in Quezon City, Metro Manila, Philippines. Acanthamoeba species were isolated from eight (16%) in 50 contact lens cases examined. We analyzed partial 18S ribosomal DNA (Rns) sequences of the eight isolates and found that the sequence differences were sufficient to distinguish the genotypes. After the isolates were genotyped, using the Basic Local Alignment Search Tool program, their phylogenetic positions relative to known Acanthamoeba isolates were determined. The model-based (GTR+Gamma+Iota) neighbor-joining, maximum likelihood, and Bayesian inference analyses, as well as the non-model-based maximum parsimony analysis were used. Results showed that of the eight isolates, six were Rns genotype T5 while two were Rns genotype T4. This present study indicates that genotype T5 is also a common contaminant in contact lens storage cases.

  20. Isolation and molecular characterization of Acanthamoeba and Balamuthia mandrillaris from combination shower units in Costa Rica.

    PubMed

    Retana-Moreira, Lissette; Abrahams-Sandí, Elizabeth; Cabello-Vílchez, Alfonso Martín; Reyes-Batlle, María; Valladares, Basilio; Martínez-Carretero, Enrique; Piñero, José E; Lorenzo-Morales, Jacob

    2014-11-01

    Free living amoebae (FLA) are ubiquitous protozoa, which may behave as parasites under certain conditions. Four genera are recognized as causal agents of infections in humans and animals: Naegleria, Sappinia, Acanthamoeba and Balamuthia. This work determines the presence of FLA in combination shower units and employs molecular biology for the characterization of isolates. The morphological analysis and partial sequencing of the 18S rDNA gene revealed the presence of Acanthamoeba genotype T4 in 30% of the units sampled. In addition to Acanthamoeba cysts, trophozoites with morphological characteristics similar to Balamuthia were identified. PCR assay using the mitochondrial 16S rRNA gene as a target confirmed the identification of the amoeba as Balamuthia mandrillaris. Up to date, this is the first report of the isolation of B. mandrillaris in Central America and the fifth report worldwide.

  1. Effects of magainins on ameba and cyst stages of Acanthamoeba polyphaga.

    PubMed Central

    Schuster, F L; Jacob, L S

    1992-01-01

    Amebic keratitis produced by Acanthamoeba spp. is an increasingly important ocular infection in extended-use contact lens wearers. Problems associated with the infection are compounded by the lack of effective and well-tolerated chemotherapeutic agents. The magainins, a group of naturally occurring and synthetic membrane-active peptide compounds, have been shown to be active in vitro against a clinical isolate of Acanthamoeba polyphaga. Two magainins tested extensively had minimal inhibitory and minimal amebicidal values of 20 and 25 micrograms/ml for magainin MSI-103 and 25 and 40 micrograms/ml for magainin MSI-94, respectively. Both amebastatic and amebicidal activities are enhanced by combining the magainins with silver nitrate (200 micrograms/ml) and/or other marginally effective antimicrobial agents. These combinations have activity against both trophic and cystic stages in the Acanthamoeba life cycle and have promise as antimicrobial agents in the treatment of amebic keratitis. PMID:1416825

  2. Degradation of immunoglobulins, protease inhibitors, and interleukin-1 by a secretory proteinase of Acanthamoeba castellanii

    PubMed Central

    Na, Byoung-Kuk; Cho, Jong-Hwa; Song, Chul-Yong; Kim, Tong-Soo

    2002-01-01

    The effect of a secretory proteinase from the pathogenic amoebae Acanthamoeba castellanii on host's defense-oriented or regulatory proteins such as immunoglobulins, interleukin-1, and protease inhibitors was investigated. The enzyme was found to degrade secretory immunoglobulin A (sIgA), IgG, and IgM. It also degraded interleukin-1α (IL-1α) and IL-1β. Its activity was not inhibited by endogenous protease inhibitors, such as α2-macroglobulin, α1-trypsin inhibitor, and α2-antiplasmin. Furthermore, the enzyme rapidly degraded those endogenous protease inhibitors as well. The degradation of host's defense-oriented or regulatory proteins by the Acanthamoeba proteinase suggested that the enzyme might be an important virulence factor in the pathogenesis of Acanthamoeba infection. PMID:12073735

  3. Acanthamoeba keratitis presenting as dendritic keratitis in a soft contact lens wearer.

    PubMed

    Yeung, Edward Yip; Huang, Samuel Chao-Ming; Tsai, Ray Jui-Fang

    2002-03-01

    Acanthamoeba keratitis is a rare cause of corneal infection in Taiwan, which can result in devastating visual outcomes. A 37-year-old woman, who wore soft contact lenses, suffered from severe pain in her left eye. Biomicroscopy revealed dendritic keratitis, radial keratoneuritis, and fine keratic precipitates on her cornea. Culture, using non-nutrient agar plate seeded with Escherichia coli, resulted in heavy growth of Acanthamoeba. The inpatient treatment, including topical neomycin-polymyxin B and metronidazole (0.5%) eyedrops, oral ketoconazole, and then oral prednisolone, successfully controlled the corneal infection. The best-corrected visual acuity was 0.9 without any evidence of recurrence of infection after 21 months of follow up. Acanthamoeba keratitis can present as dendritic keratitis, which mimics herpes simplex infection, thus, delays appropriate treatment. Early diagnosis and judicious treatment are essential for restoring the vision and avoiding the subsequent need of penetrating keratoplasty.

  4. [Distribution and compensation mechanism of aberrations between anterior and posterior surface of the cornea in myopia and myopic astigmatism eyes].

    PubMed

    Li, X J; Wang, Y; Wu, Y N; Wu, W J; Yu, C J; Xu, L L

    2016-11-11

    Objective: To study the compensation mechanism of aberrations between anterior and posterior corneal surface, and to investigate the correlations between corneal aberrations and K values, intraocular pressure and central corneal thickness. Methods: One hundred and sixty-one subjects (300 eyes) with myopia and myopic astigmatism were recruited randomly. Corneal aberrations (anterior, posterior and total) in three different optical zones (2 mm, 4 mm, and 6 mm) centered on the corneal vertex were assessed with a corneal topographer (Sirius). We also calculated compensation factors (CFs) as a measure of the relative efficiency of the aberration compensation mechanism. Astigmatism was divided into corneal astigmatism and non-corneal astigmatism. Mann-Whitney U test was utilized for the comparison of different aberrations and CFs between the two astigmatism groups. Spearman correlation was applied to analyze the correlations between corneal aberrations and K values, intraocular pressure and central corneal thickness. Results: As the order of the aberrations elevated from the second to the seventh, the RMS values decreased significantly. The larger the optical zone, the greater the values of aberrations and the lower the percentage of the compensation mechanism among all the Zernike terms. At the same time, as the order of the aberrations increased, the predominance of the compensation mechanism was increasingly obvious. Slight compensation of spherical aberration (Z4(0)) was observed in the peripheral. In the center of the analyzing zones (2 mm), compensation mechanism represented in coma (Z3(±1)), and it disappeared in the peripheral. We detected slight compensation of the corneal astigmatism (Z2(±2)) in the surrounding zone, although the compensation factors were closed to zero in diverse optical zones. Superposition was found in trefoil (Z3(±3)) straightly. Nevertheless, a tendency towards compensation was discovered with the enlargement of the optical zones. And

  5. Risk factors for acanthamoeba keratitis in contact lens users: a case-control study.

    PubMed Central

    Radford, C. F.; Bacon, A. S.; Dart, J. K.; Minassian, D. C.

    1995-01-01

    OBJECTIVE--To investigate reasons for an increase in cases of Acanthamoeba keratitis related to contact lenses. DESIGN--Case-control study. Cases were contact lens related acanthamoeba keratitis patients treated between 1 September 1989 and 31 August 1992. Controls were lens users without lens related disease who presented as new patients to the casualty department from 1 March 1992 to 31 August 1992. All subjects completed a questionnaire detailing lens use and hygiene practices. SETTING--Eye hospital. SUBJECTS--35 cases with acanthamoeba keratitis and 378 controls. MAIN OUTCOME MEASURES--Relative risks comparing different contact lens types, socioeconomic classification, age, sex, lens use, lens wearing experience, hygiene compliance, and hygiene systems. RESULTS--The crude relative risk for developing acanthamoeba keratitis with the use of daily wear disposable lenses was 49.45 (95% confidence interval 6.53 to 2227; P < 0.001) compared with conventional soft lenses (the referent). Multivariable analysis showed that this increased risk could be largely attributed to lack of disinfection (relative risk 55.86 (10 to 302); P < 0.001) and use of chlorine based disinfection (14.63 (2.8 to 76); P = 0.001) compared with other chemical systems (the referent). None of the other outcome measures showed a significant association. CONCLUSIONS--Both failure to disinfect daily wear soft contact lenses and the use of chlorine release lens disinfection systems, which have little protective effect against the organism, are major risk factors for acanthamoeba keratitis. These risks have been particularly common in disposable lens use. Over 80% of acanthamoeba keratitis could be avoided by the use of lens disinfection systems that are effective against the organism. PMID:7787645

  6. Validation of real-time PCR for laboratory diagnosis of Acanthamoeba keratitis.

    PubMed

    Thompson, Paul P; Kowalski, Regis P; Shanks, Robert M Q; Gordon, Y Jerold

    2008-10-01

    Confirmation of Acanthamoeba keratitis by laboratory diagnosis is the first step in the treatment of this vision-threatening disease. Two real-time PCR TaqMan protocols (the Rivière and Qvarnstrom assays) were developed for the detection of genus-specific Acanthamoeba DNA but lacked clinical validation. We have adapted these assays for the Cepheid SmartCycler II system (i) by determining their real-time PCR limits of detection and amplification efficiencies, (ii) by determining their ability to detect trophozoites and cysts, and (iii) by testing a battery of positive and negative samples. We also examined the inhibitory effects of a number of commonly used topical ophthalmic drugs on real-time PCR. The results of the real-time PCR limit of detection and amplification efficiency of the Rivière and Qvarnstrom assays were 11.3 DNA copies/10 microl and 94% and 43.8 DNA copies/10 microl and 92%, respectively. Our extraction protocol enabled us to detect 0.7 Acanthamoeba cysts/10 microl and 2.3 Acanthamoeba trophozoites/10 microl by both real-time PCR assays. The overall agreement between the assays was 97.0%. The clinical sensitivity and specificity of both real-time PCR assays based on culture were 100% (7 of 7) and 100% (37 of 37), respectively. Polyhexamethylene biguanide was the only topical drug that demonstrated PCR inhibition, with a minimal inhibitory dilution of 1/640 and an amplification efficiency of 72.7%. Four clinical samples were Acanthamoeba culture negative and real-time PCR positive. Our results indicate that both real-time PCR assays could be used to diagnose Acanthamoeba keratitis. Polyhexamethylene biguanide can inhibit PCR, and we suggest that specimen collection occur prior to topical treatment to avoid possible false-negative results.

  7. Acanthamoeba strains lose their abilities to encyst synchronously upon prolonged axenic culture.

    PubMed

    Köhsler, Martina; Leitsch, David; Fürnkranz, Ursula; Duchêne, Michael; Aspöck, Horst; Walochnik, Julia

    2008-04-01

    To evaluate the influence of prolonged axenic culture on the encystment capacity of Acanthamoeba spp., the encystment potential of four closely related Acanthamoeba strains, subcultured axenically for different periods of time, was evaluated comparing five encystment media. Media with more alkaline pH values were slightly more effective; however, the composition of the respective encystment medium had only limited influence on the encystment potential, while a strong correlation of losses in encystment potential and times strains had been cultured axenically was demonstrated. Furthermore, our results indicate that losses in encystment potential occur shortly after transfer into axenic culture to remain constant over many years.

  8. Killing of diverse eye pathogens (Acanthamoeba spp., Fusarium solani, and Chlamydia trachomatis) with alcohols

    PubMed Central

    2017-01-01

    Background Blindness is caused by eye pathogens that include a free-living protist (Acanthamoeba castellanii, A. byersi, and/or other Acanthamoeba spp.), a fungus (Fusarium solani), and a bacterium (Chlamydia trachomatis). Hand-eye contact is likely a contributor to the spread of these pathogens, and so hand washing with soap and water or alcohol–based hand sanitizers (when water is not available) might reduce their transmission. Recently we showed that ethanol and isopropanol in concentrations present in hand sanitizers kill walled cysts of Giardia and Entamoeba, causes of diarrhea and dysentery, respectively. The goal here was to determine whether these alcohols might kill infectious forms of representative eye pathogens (trophozoites and cysts of Acanthamoeba, conidia of F. solani, or elementary bodies of C. trachomatis). Methodology/Principal findings We found that treatment with 63% ethanol or 63% isopropanol kills >99% of Acanthamoeba trophozoites after 30 sec exposure, as shown by labeling with propidium iodide (PI) and failure to grow in culture. In contrast, Acanthamoeba cysts, which contain cellulose fibers in their wall, are relatively more resistant to these alcohols, particularly isopropanol. Depending upon the strain tested, 80 to 99% of Acanthamoeba cysts were killed by 63% ethanol after 2 min and 95 to 99% were killed by 80% ethanol after 30 sec, as shown by PI labeling and reduced rates of excystation in vitro. Both ethanol and isopropanol (63% for 30 sec) kill >99% of F. solani conidia, which have a wall of chitin and glucan fibrils, as demonstrated by PI labeling and colony counts on nutrient agar plates. Both ethanol and isopropanol (63% for 60 sec) inactivate 96 to 99% of elementary bodies of C. trachomatis, which have a wall of lipopolysaccharide but lack peptidoglycan, as measured by quantitative cultures to calculate inclusion forming units. Conclusions/Significance In summary, alcohols kill infectious forms of Acanthamoeba, F. solani, and

  9. Propylene glycol and contact-lens solutions containing this diol induce pseudocyst formation in acanthamoebae.

    PubMed

    Kliescikova, Jarmila; Kulda, Jaroslav; Nohynkova, Eva

    2011-01-01

    Propylene glycol used as an ophthalmic demulcent in certain contact-lens care systems has been included recently among factors responsible for increasing Acanthamoeba keratitis. In this study, we provide evidence that propylene glycol as well as examined contact-lens solutions containing it induce rapid differentiation of acanthamoebae into pseudocysts. The partial resistance of the pseudocysts and their reversibility to viable trophozoites even after 24-h exposure to the contact-lens solutions indicate a potential risk of infection to contact-lens users.

  10. Random amplified polymorphic DNA profiles as a tool for the identification of Acanthamoeba divionensis.

    PubMed

    Ortega-Rivas, Antonio; Lorenzo-Morales, Jacob; Alonso, Violeta; Abreu, Néstor J; Foronda, Pilar; del Castillo, Antonio; Valladares, Basilio

    2003-08-01

    In the present study, we demonstrated the Random Amplified Polymorphism DNA (RAPD) diagnostic validity. In our study, we have analyzed RAPD profiles searching for characteristic and useful bands for Acanthamoeba diagnosis at the species level. We found a distinctive 370-bp band in A. divionensis RAPD patterns, using the OPC14 primer (TGCGTGCTTG) (Operon Technologies, Inc., Alameda, CA). The band specificity was confirmed by hybridization, using it as a probe, against all OPC14 amplifications from 10 different Acanthamoeba species. Once we sequenced this band, we used it to design a specific primer pair which showed positive amplification only in A. divionensis isolates.

  11. Monomeric Acanthamoeba myosins I support movement in vitro.

    PubMed

    Albanesi, J P; Fujisaki, H; Hammer, J A; Korn, E D; Jones, R; Sheetz, M P

    1985-07-25

    Acanthamoeba myosins IA and IB were found to have molecular weights of 159,000 and 150,000 and Stokes radii of 6.2 and 5.9 nm, respectively. Both enzymes have frictional ratios of 1.7. Myosin IA consists of 22% alpha-helix, 32% beta-structure, and 46% unordered structure, while myosin IB is 16% alpha-helix, 46% beta-structure, and 38% unordered. Both myosins remain monomolecular under conditions in which other myosins form filaments. Beads coated with myosin IA or IB move unidirectionally on actin cables of Nitella. Movement requires ATP and phosphorylation of the myosin I heavy chain which is also required for actin-activated Mg2+-ATPase activity. Movement is inhibited by myosin I antiserum that inhibits actin-activated ATPase activity. These studies establish that these nonfilamentous, monomolecular myosins with single heavy chains of 130,000 and 125,000 daltons (IA and IB, respectively) can support actin-dependent movement analogous to that supported by filamentous myosins.

  12. Legionella pneumophila prevents proliferation of its natural host Acanthamoeba castellanii

    PubMed Central

    Mengue, Luce; Régnacq, Matthieu; Aucher, Willy; Portier, Emilie; Héchard, Yann; Samba-Louaka, Ascel

    2016-01-01

    Legionella pneumophila is a ubiquitous, pathogenic, Gram-negative bacterium responsible for legionellosis. Like many other amoeba-resistant microorganisms, L. pneumophila resists host clearance and multiplies inside the cell. Through its Dot/Icm type IV secretion system, the bacterium injects more than three hundred effectors that modulate host cell physiology in order to promote its own intracellular replication. Here we report that L. pneumophila prevents proliferation of its natural host Acanthamoeba castellanii. Infected amoebae could not undergo DNA replication and no cell division was observed. The Dot/Icm secretion system was necessary for L. pneumophila to prevent the eukaryotic proliferation. The absence of proliferation was associated with altered amoebal morphology and with a decrease of mRNA transcript levels of CDC2b, a putative regulator of the A. castellanii cell cycle. Complementation of CDC28-deficient Saccharomyces cerevisiae by the CDC2b cDNA was sufficient to restore proliferation of CDC28-deficient S. cerevisiae and suggests for the first time that CDC2b from A. castellanii could be functional and a bona fide cyclin-dependent kinase. Hence, our results reveal that L. pneumophila impairs proliferation of A. castellanii and this effect could involve the cell cycle protein CDC2b. PMID:27805070

  13. Acanthamoeba castellanii: structural basis of the cytopathic mechanisms.

    PubMed

    González-Robles, Arturo; Castañón, Guadalupe; Cristóbal-Ramos, Ana Ruth; Lázaro-Haller, Amparo; Omaña-Molina, Maritza; Bonilla, Patricia; Martínez-Palomo, Adolfo

    2006-11-01

    In this study we report observations on the structural mechanisms of the cytopathic effect of Acanthamoeba castellanii trophozoites on cultured MDCK cell monolayers. Co-incubations were carried out for a maximum of 24h. The first evidence of damage to the cell monolayer was detected by measuring the transepithelial resistance of cell monolayers that interacted with the amoebae. At 6h, transepithelial resistance diminished to 51% and amoebae required 5-6h to produce evidence of structural injury at the light microscopy level. Following 12h of incubation, the cell monolayer was severely damaged. After making intimate contact with the surface of target cells, trophozoites detached cells from the substrate, lysed and by means of food-cups ingested the damaged cells. There was no morphological evidence of modifications in MDCK cell membranes, membrane fusion or junction formation between the amoeba and host plasma membrane. The lytic capacity of the amoebas appears to be the result of cytotoxic factors secreted by the amoebae since, when monolayers were incubated with conditioned medium, there was also a decrease in the transepithelial resistance. Besides, mechanical injury produced by the attachment and movement of the trophozoites may contribute to the disruption of the cell monolayer. As in other pathogenic amoebae, the cytopathic action of A. castellanii on the cell monolayers can subjectively be separated into four stages: adhesion, cytolysis, phagocytosis, and intracellular degradation.

  14. Shiga toxins decrease enterohaemorrhagic Escherichia coli survival within Acanthamoeba castellanii.

    PubMed

    Chekabab, Samuel M; Daigle, France; Charette, Steve J; Dozois, Charles M; Harel, Josée

    2013-07-01

    Enterohaemorrhagic Escherichia coli (EHEC) are zoonotic pathogens transmitted to humans through contaminated water or bovine products. One of the strategies used by pathogenic bacteria to survive in aquatic environments is using free-living amoebae as hosts. Acanthamoeba castellanii is an amoeba known to host several waterborne pathogens. This study investigates the survival of EHEC with A. castellanii, which could contribute to its spread and transmission to humans. We used a gentamicin protection assay as well as fluorescence and electron microscopy to monitor the intra-amoebae survival of EHEC O157:H7 over 24 h. The results showed that EHEC were able to survive within A. castellanii and that this survival was reduced by Shiga toxins (Stx) produced by EHEC. A toxic effect mediated by Stx was demonstrated by amoebae mortality and LDH release during co-culture of EHEC and amoeba. This work describes the ability of EHEC to survive within A. castellanii, and this host-pathogen interaction is partially controlled by the Stx. Thus, this ubiquitous amoeba could represent an environmental niche for EHEC survival and transmission.

  15. Growth of Legionella pneumophila in Acanthamoeba castellanii enhances invasion.

    PubMed Central

    Cirillo, J D; Falkow, S; Tompkins, L S

    1994-01-01

    Legionella pneumophila is considered to be a facultative intracellular parasite. Therefore, the ability of these bacteria to enter, i.e., invade, eukaryotic cells is expected to be a key pathogenic determinant. We compared the invasive ability of bacteria grown under standard laboratory conditions with that of bacteria grown in Acanthamoeba castellanii, one of the protozoan species that serves as a natural host for L. pneumophila in the environment. Amoeba-grown L. pneumophila cells were found to be at least 100-fold more invasive for epithelial cells and 10-fold more invasive for macrophages and A. castellanii than were L. pneumophila cells grown on agar. Comparison of agar- and amoeba-grown L. pneumophila cells by light and electron microscopy demonstrated dramatic differences in the morphology and structure of the bacteria. Analyses of protein expression in the two strains of bacteria suggest that these phenotypic differences may be due to the expression of new proteins in amoeba-grown L. pneumophila cells. In addition, the amoeba-grown bacteria were found to enter macrophages via coiling phagocytosis at a higher frequency than agar-grown bacteria did. Replication of L. pneumophila in protozoans present in domestic water supplies may be necessary to produce bacteria that are competent to enter mammalian cells and produce human disease. Images PMID:8039895

  16. Survival and Growth of Francisella tularensis in Acanthamoeba castellanii

    PubMed Central

    Abd, Hadi; Johansson, Thorsten; Golovliov, Igor; Sandström, Gunnar; Forsman, Mats

    2003-01-01

    Francisella tularensis is a highly infectious, facultative intracellular bacterium which causes epidemics of tularemia in both humans and mammals at regular intervals. The natural reservoir of the bacterium is largely unknown, although it has been speculated that protozoa may harbor it. To test this hypothesis, Acanthamoeba castellanii was cocultured with a strain of F. tularensis engineered to produce green fluorescent protein (GFP) in a nutrient-rich medium. GFP fluorescence within A. castellanii was then monitored by flow cytometry and fluorescence microscopy. In addition, extracellular bacteria were distinguished from intracellular bacteria by targeting with monoclonal antibodies. Electron microscopy was used to determine the intracellular location of F. tularensis in A. castellanii, and viable counts were obtained for both extracellular and intracellular bacteria. The results showed that many F. tularensis cells were located intracellularly in A. castellanii cells. The bacteria multiplied within intracellular vacuoles and eventually killed many of the host cells. F. tularensis was found in intact trophozoites, excreted vesicles, and cysts. Furthermore, F. tularensis grew faster in cocultures with A. castellanii than it did when grown alone in the same medium. This increase in growth was accompanied by a decrease in the number of A. castellanii cells. The interaction between F. tularensis and amoebae demonstrated in this study indicates that ubiquitous protozoa might be an important environmental reservoir for F. tularensis. PMID:12514047

  17. The interaction of Acanthamoeba castellanii cysts with macrophages and neutrophils.

    PubMed

    Hurt, Michael; Proy, Vincent; Niederkorn, Jerry Y; Alizadeh, Hassan

    2003-06-01

    Acanthamoeba castellanii, a free-living amoeba, causes a sight-threatening form of keratitis. Even after extensive therapies, corneal damage can be severe, often requiring corneal transplantation to restore vision. However, A. castellanii cysts are not eliminated from the conjunctiva and stroma of humans and can excyst, resulting in infection of the corneal transplant. The aim of this study was to determine whether elements of the innate immune apparatus, neutrophils and macrophages, were capable of detecting and eliminating A. castellanii cysts and to examine the mechanism by which they kill the cysts. Results show that neither innate immune cell is attracted chemotactically to intact cysts, yet both were attracted to lysed cysts. Both macrophages and neutrophils were capable of killing significant numbers of cysts, yet neutrophils were 3-fold more efficient than macrophages. Activation of macrophages with lipopolysaccharide and interferon-gamma did not increase their cytolytic ability. Conditioned medium isolated from macrophages did not lyse the cysts; however, prevention of phagocytosis by cytochalasin D inhibited 100% of macrophage-mediated killing of the cysts. Conditioned medium from neutrophils did kill significant numbers of the cysts, and this killing was blocked by quercetin, a potent inhibitor of myeloperoxidase (MPO). These results indicate that neither macrophages nor neutrophils are chemoattracted to intact cysts, yet both are capable of killing the cysts. Macrophages killed the cysts by phagocytosis, whereas neutrophils killed cysts through the secretion of MPO.

  18. Staphylococcus aureus exhibit similarities in their interactions with Acanthamoeba and ThP1 macrophage-like cells.

    PubMed

    Cardas, Mihaela; Khan, Naveed Ahmed; Alsam, Selwa

    2012-12-01

    Staphylococcus aureus is a leading cause of nosocomial infections. Haematogenous spread is a pre-requisite but it is not clear how S. aureus survive the onslaught of macrophages. Acanthamoeba is a protozoan pathogen that is remarkably similar to macrophages, particularly in their cellular structure (morphological and ultra-structural features), molecular motility, biochemical physiology, ability to capture prey by phagocytosis and interactions with microbial pathogens. Thus, we hypothesize that S. aureus exhibit similarities in their interactions with Acanthamoeba and ThP1 macrophage-like cells. Here, we studied interactions of methicillin-sensitive S. aureus (MSSA), methicillin-resistant S. aureus (MRSA) and Staphylococcus epidermidis (SE) with Acanthamoeba castellanii belonging to the T4 genotype and macrophage-like cells (ThP1). The findings revealed that both MRSA and MSSA exhibited similarities in their binding/association and invasion of A. castellanii and ThP1 cells. Long-term incubation showed that MRSA and MSSA can survive intracellularly of both Acanthamoeba and ThP1 cells. Overall, these findings suggest that Acanthamoeba exhibit similar characteristics with ThP1 macrophage-like cells in their interaction with MRSA and MSSA. Additionally it was shown that bacteria survive inside Acanthamoeba during the encystment process as evidenced by bacterial recovery from mature cysts. Given that Acanthamoeba cysts are airborne, these findings suggest that cysts may act as "Trojan horse" to help spread MRSA to susceptible hosts.

  19. 3D multifocus astigmatism and compressed sensing (3D MACS) based superresolution reconstruction

    PubMed Central

    Huang, Jiaqing; Sun, Mingzhai; Gumpper, Kristyn; Chi, Yuejie; Ma, Jianjie

    2015-01-01

    Single molecule based superresolution techniques (STORM/PALM) achieve nanometer spatial resolution by integrating the temporal information of the switching dynamics of fluorophores (emitters). When emitter density is low for each frame, they are located to the nanometer resolution. However, when the emitter density rises, causing significant overlapping, it becomes increasingly difficult to accurately locate individual emitters. This is particularly apparent in three dimensional (3D) localization because of the large effective volume of the 3D point spread function (PSF). The inability to precisely locate the emitters at a high density causes poor temporal resolution of localization-based superresolution technique and significantly limits its application in 3D live cell imaging. To address this problem, we developed a 3D high-density superresolution imaging platform that allows us to precisely locate the positions of emitters, even when they are significantly overlapped in three dimensional space. Our platform involves a multi-focus system in combination with astigmatic optics and an ℓ1-Homotopy optimization procedure. To reduce the intrinsic bias introduced by the discrete formulation of compressed sensing, we introduced a debiasing step followed by a 3D weighted centroid procedure, which not only increases the localization accuracy, but also increases the computation speed of image reconstruction. We implemented our algorithms on a graphic processing unit (GPU), which speeds up processing 10 times compared with central processing unit (CPU) implementation. We tested our method with both simulated data and experimental data of fluorescently labeled microtubules and were able to reconstruct a 3D microtubule image with 1000 frames (512×512) acquired within 20 seconds. PMID:25798314

  20. Transmissive liquid-crystal device correcting primary coma aberration and astigmatism in laser scanning microscopy

    NASA Astrophysics Data System (ADS)

    Tanabe, Ayano; Hibi, Terumasa; Ipponjima, Sari; Matsumoto, Kenji; Yokoyama, Masafumi; Kurihara, Makoto; Hashimoto, Nobuyuki; Nemoto, Tomomi

    2016-03-01

    Laser scanning microscopy allows 3D cross-sectional imaging inside biospecimens. However, certain aberrations produced can degrade the quality of the resulting images. We previously reported a transmissive liquid-crystal device that could compensate for the predominant spherical aberrations during the observations, particularly in deep regions of the samples. The device, inserted between the objective lens and the microscope revolver, improved the image quality of fixed-mouse-brain slices that were observed using two-photon excitation laser scanning microscopy, which was originally degraded by spherical aberration. In this study, we developed a transmissive device that corrects primary coma aberration and astigmatism, motivated by the fact that these asymmetric aberrations can also often considerably deteriorate image quality, even near the sample surface. The device's performance was evaluated by observing fluorescent beads using single-photon excitation laser scanning microscopy. The fluorescence intensity in the image of the bead under a cover slip tilted in the y-direction was increased by 1.5 times after correction by the device. Furthermore, the y- and z-widths of the imaged bead were reduced to 66% and 65%, respectively. On the other hand, for the imaged bead sucked into a glass capillary in the longitudinal x-direction, correction with the device increased the fluorescence intensity by 2.2 times compared to that of the aberrated image. In addition, the x-, y-, and z-widths of the bead image were reduced to 75%, 53%, and 40%, respectively. Our device successfully corrected several asymmetric aberrations to improve the fluorescent signal and spatial resolution, and might be useful for observing various biospecimens.

  1. Occurrence of pathogenic Acanthamoeba genotypes in nasal swabs of cancer patients in Iran.

    PubMed

    Memari, Fatemeh; Niyyati, Maryam; Haghighi, Ali; Seyyed Tabaei, Seyyed Javad; Lasjerdi, Z

    2015-05-01

    Incidences of Acanthamoeba granulomatous encephalitis (AGE) have been increased due to a rise in the number of high-risk people, such as immunodeficient patients. Indeed, immunosuppress situation can render the patient in acquiring opportunistic Acanthamoeba infections. In this study, analysis was carried out to verify the presence of free-living amoebae of Acanthamoeba genus in nasal swabs of cancer patients in hospitals of Tehran, Iran. Detection of isolates was based on morphotyping and PCR sequencing of the Diagnostic Fragment 3 (DF3) to identify strains at the genotype level. In addition, the pathogenic potential of the isolates was assayed using temperature and osmotolerance assays. The obtained results revealed that nine isolated strains belonging to T4 genotype-exhibited pathogenic potential. After sequencing, genotype T4 was found to be the most common one in the samples included in this study. Genotype T3 and T5 were also identified. To the best of our knowledge, this is the first study on the typing of Acanthamoeba strains at the genotype level in cancer patients in Iran and worldwide.

  2. Isolation and identification of Acanthamoeba spp. from thermal swimming pools and spas in Southern Brazil.

    PubMed

    Fabres, Laura Fuhrich; Rosa Dos Santos, Sayonara Peixoto; Benitez, Lisianne Brittes; Rott, Marilise Brittes

    2016-03-01

    Free-living amoebae (FLA) are widely distributed in soil and water. A few number of them are implicated in human disease: Acanthamoeba spp., Naegleria fowleri, Balamuthia mandrillaris and Sappinia diploidea. Species of Acanthamoeba can cause keratitis and brain infections. In this study, 72 water samples were taken from both hot tubs and thermal swimming pools in the city of Porto Alegre, RS, Brazil, to determine the presence of Acanthamoeba in the water as well as perform the phenotypic and genotypic characterization of the isolates. The identification of the isolates was based on the cysts morphology and PCR amplification using genus-specific oligonucleotides. When the isolates were submitted to PCR reaction only 8 were confirmed as belonging to the genus Acanthamoeba. The sequences analysis when compared to the sequences in the GenBank, showed genotype distribution in group T3 (12,5%), T5 (12,5%), T4 (25%) and T15 (50%). The results of this study confirmed the presence of potentially pathogenic isolates of free living amoebae in hot swimming pool and spas which can present risks to human health.

  3. Genotyping of potentially pathogenic Acanthamoeba strains isolated from nasal swabs of healthy individuals in Peru.

    PubMed

    Cabello-Vílchez, Alfonso Martín; Martín-Navarro, Carmen María; López-Arencibia, Atteneri; Reyes-Batlle, María; González, Ana C; Guerra, Humberto; Gotuzzo, Eduardo; Valladares, Basilio; Piñero, José E; Lorenzo-Morales, Jacob

    2014-02-01

    Free Living Amoebae (FLA) of Acanthamoeba genus are widely distributed in the environment and can be found in the air, soil and water; and have also been isolated from air-conditioning units. In humans, they are causative agents of a sight-threating infection of the cornea, Acanthamoeba keratitis (AK) and a fatal infection of the central nervous system known as Granulomatous Amoebic Encephalitis (GAE). In this study, a survey was conducted in order to determine the presence and pathogenic potential of free-living amoebae of Acanthamoeba genus in nasal swabs from individuals in two regions of Peru. Identification of isolates was based on cyst morphology and PCR-sequencing of the Diagnostic Fragment 3 to identify strains at the genotype level. The pathogenic potential of the isolates was also assayed using temperature and osmotolerance assays and extracellular proteases zymograms. The obtained results revealed that all isolated strains exhibited pathogenic potential. After sequencing the highly variable DF3 (Diagnostic Fragment 3) region in the 18S rRNA gene as previously described, genotype T4 was found to be the most common one in the samples included in this study but also genotype T15 was identified. To the best of our knowledge, this is the first study on the characterization of Acanthamoeba strains at the genotype level and the first report of genotype T4 and T15 in Peru.

  4. Microarray and KOG analysis of Acanthamoeba healyi genes up-regulated by mouse-brain passage.

    PubMed

    Moon, Eun-Kyung; Xuan, Ying-Hua; Kong, Hyun-Hee

    2014-08-01

    Long-term cultivation in a laboratory could reduce the virulence of Acanthamoeba. To identify virulence factors of Acanthamoeba, the authors compared the transcription profiles of long-term cultivated Acanthamoeba healyi (OLD) and three times mouse-brain passaged A. healyi (MBP) using microarray analysis and eukaryotic orthologous group (KOG) assignments. Microarray analysis revealed that 601 genes were up-regulated by mouse-brain passage. The results of real-time PCR of 8 randomly selected genes up-regulated in the MBP strain confirmed microarray analysis findings. KOG assignments showed relatively higher percentages of the MBP strain up-regulated genes in T article (signal transduction mechanism), O article (posttranslational modification, protein turnover, chaperones), C article (energy production and conversion), and J article (translation, ribosomal structure and biogenesis). In particular, the MBP strain showed higher expressions of cysteine protease and metalloprotease. A comparison of KOG assignments by microarray analysis and previous EST (expressed sequence tags) analysis showed similar populations of up-regulated genes. These results provide important information regarding the identification of virulence factors of pathogenic Acanthamoeba.

  5. Efficacy of Korean Multipurpose Contact Lens Disinfecting Solutions against Acanthamoeba castellanii

    PubMed Central

    Moon, Eun-Kyung; Park, Hye-Ryun; Quan, Fu-Shi; Kong, Hyun-Hee

    2016-01-01

    Acanthamoeba keratitis has been increasing in recent years. Main risk factors are contact lens wear and their cleaning solutions. Most contact lens wearers use multipurpose disinfecting solutions (MPDS) for cleansing and disinfecting microorganisms because of its convenience. We determined amoebicidal effects of MPDS made in Korea and their cytotoxicity on human corneal epithelium cells. Fifteen commercial MPDS (A to O) were tested for their amoebicidal effects on Acanthamoeba castellanii trophozoites and cysts by using a most probable number (MPN) technique. Among them, 7 kinds of MPDS showed little or no amoebicidal effects for 24 hr exposure. Solutions A, B, G, H, L, and O showed positive amoebicidal effects, and solutions M and N killed almost all trophozoites and cysts after 24 hr exposure. However, 50%-N solution showed 56% cytotoxicity on human corneal epithelial cells within 4 hr exposure, and 50%-O solution also showed 62% cytotoxicity on human cells within 4 hr exposure. Solution A did not show any cytotoxicity on human cells. These results revealed that most MPDS made in Korea were ineffective to kill Acanthamoeba. The solutions having amoebicidal activity also showed high levels of cytotoxicity on human corneal epithelial cells. New formulations for improved MPDS that are amoebicidal but safe for host cells are needed to prevent Acanthamoeba keratitis. PMID:28095653

  6. Rapid diagnosis of acanthamoeba keratitis using non-nutrient agar with a lawn of E. coli

    PubMed Central

    2013-01-01

    Background A patient presented with a corneal foreign body in his only eye. He was treated with prophylactic antibiotics and sent home, but deteriorated. Findings He returned to the hospital 5 days later, and on slit-lamp examination, there was ciliary injection, corneal oedema and a 1 mm × 1 mm corneal abscess with mild anterior uveitis. Corneal scrapings were taken for culture on a non-nutrient agar with a lawn of Escherichia coli, on chocolate agar and on blood agar. He was treated with fortified gentamicin and cefazolin drops. He improved and was discharged 4 days after admission. On day 5, the culture results showed acanthamoeba. He was brought back to the hospital and treated with hourly chlorhexidine drops, ofloxacin six times daily and neomycin/dexamethasone drops once daily. On day 7, he was discharged to continue treatment at home, at which time his visual acuity in that eye was 6/9, and slit-lamp examination showed punctate keratitis and a stromal opacity with mild peripheral infiltration. Conclusions Culture on non-nutrient agar with a lawn of E. coli is a rapid, reliable and less invasive alternative to corneal biopsy for the diagnosis of acanthamoeba infection. We suggest using this method where acanthamoeba is suspected. Owing to the risk of corneal abscess, orthokeratology should be avoided in an amblyopic patient or an only eye. Acanthamoeba infection may be masked by other eye diseases. PMID:23514313

  7. Acanthamoeba polyphaga is a possible host for Vibrio cholerae in aquatic environments.

    PubMed

    Sandström, Gunnar; Saeed, Amir; Abd, Hadi

    2010-09-01

    Acanthamoeba is a genus of free-living amoebae found to be able to host many bacterial species living in the environment. Acanthamoebae and Vibrio cholerae are found in the aquatic environments of cholera endemic areas. Previously it has been shown that V. cholerae O1 and O139 can survive and grow in Acanthamoeba castellanii. The aim of this study was to examine the ability of Acanthamoeba polyphaga to host V. cholerae O1 and O139. The interaction between A. polyphaga and V. cholerae strains was studied by means of viable amoeba cell counts and viable count of the bacteria in the absence and presence of amoebae. The viable count of intracellularly growing bacteria was estimated by utilizing gentamicin assay. Electron microscopy was used to determine the localization of V. cholerae inside A. polyphaga. The results showed that A. polyphaga enhanced growth and survival of V. cholerae, which grew and survived inside the amoeba cells for 2weeks. The electron microscopy showed that A. polyphaga hosted intracellular V. cholerae localized in the vacuoles of amoeba cell. Neither the presence of V. cholerae together with A. polyphaga nor the intracellular localization of the bacteria inhibited growth and survival of A. polyphaga. The outcome of the interaction between these microorganisms may support strongly the role of A. polyphaga as host for V. cholerae O1 and O139.

  8. Isolation and characterization of Acanthamoeba spp. from air-conditioners in Kuala Lumpur, Malaysia.

    PubMed

    Chan, Li-Li; Mak, Joon-Wah; Low, Yoon-Tong; Koh, Thuan-Tzen; Ithoi, Init; Mohamed, Shar Mariam

    2011-01-01

    During a study on the quality of the indoor environment, Acanthamoeba spp. were detected in 20 out of 87 dust samples collected from air-conditioners installed in a four-story campus building located in Kuala Lumpur, Malaysia. Twenty-one cloned Acanthamoeba isolates designated as IMU1 to IMU21 were established from the positive primary cultures. Five species were identified from the 16 isolates according to the morphological criteria of Pussard and Pons; i.e. A. castellanii, A. culbertsoni, A. griffini, A. hatchetti and A. polyphaga. Species identities for the remaining five isolates (IMU4, IMU5, IMU15, IMU20 and IMU21), however, could not be determined morphologically. At genotypic characterization, these isolates were placed into T3 (IMU14); T5 (IMU16 and IMU17) and T4 (all the remaining isolates). To predict the potential pathogenicity of these Acanthamoeba isolates, thermo- and osmotolerance tests were employed; many isolates were predicted as potential human pathogens based on the outcome of these tests. This is the first time potentially pathogenic Acanthamoeba have been isolated from air-conditioners in Malaysia.

  9. Identification of the 18S-ribosomal-DNA genotypes of Acanthamoeba isolates from the Philippines.

    PubMed

    Rivera, W L; Adao, D E V

    2008-12-01

    Cyst morphology has been commonly used to identify the free-living amoeba Acanthamoeba to subgenus level. A more accurate and consistent method, based on the sequence analysis of the gene coding for the amoeba's small-subunit ribosomal RNA (Rns), has, however, been developed. There have been no attempts to identify the Acanthamoeba genotypes circulating in the Philippines. In this study, therefore, the ASA.S1 region of the Rns gene from 17 Acanthamoeba isolates, collected from soil, water and contact-lens storage cases in different regions of the Philippines, was sequenced. After the isolates were genotyped, using the BLAST program, their phylogenetic positions relative to known Acanthamoeba isolates were determined. For this, the model-based (GTR + Gamma) neighbour-joining, maximum-likelihood and Bayesian-inference analyses and the non-model-based maximum-parsimony analysis were used. All but two of the isolates were identified as the T5 or T4 genotypes, which are probably common in soil, water and contact-lens cases across the Philippines. The only other genotypes identified were T15 (as a single isolate from a contact-lens case) and T3 (as a single soil isolate).

  10. Acanthamoeba Encephalitis: Isolation of Genotype T1 in Mycobacterial Liquid Culture Medium

    PubMed Central

    Azzam, Rula; Badenoch, Paul R.; Francis, Michelle J.; Fernandez, Charles; Adamson, Penelope J.; Dendle, Claire; Woolley, Ian; Robson, Jenny; Korman, Tony M.

    2014-01-01

    We report a case of Acanthamoeba encephalitis diagnosed from an antemortem brain biopsy specimen, where the organism was first isolated in mycobacterial liquid medium and first identified by using a sequence generated by a commercial panfungal sequencing assay. We correlate susceptibility results with clinical outcome. PMID:25502534

  11. Viability of Listeria monocytogenes in co-culture with Acanthamoeba spp.

    PubMed

    Akya, Alisha; Pointon, Andrew; Thomas, Connor

    2009-10-01

    Listeria monocytogenes is a human pathogen, ubiquitous in the environment, and can grow and survive under a wide range of environmental conditions. It contaminates foods via raw materials or food-processing environments. However, the current knowledge of its ecology and, in particular, the mode of environmental survival and transmission of this intracellular pathogen remains limited. Research has shown that several intracellular pathogens are able to survive or replicate within free-living amoebae. To examine the viability of L. monocytogenes in interaction with Acanthamoeba spp., bacteria were co-cultured with three freshly isolated amoebae, namely Acanthamoeba polyphaga, Acanthamoeba castellanii and Acanthamoeba lenticulata. The survival of bacteria and amoebae was determined using culture techniques and microscopy. Under the experimental conditions used, all amoebae were able to eliminate bacteria irrespective of the hly gene. Bacteria did not survive or replicate within amoeba cells. However, extra-amoebic bacteria grew saprophytically on materials released from amoebae, which may play an important role in the survival of bacteria under extreme environmental conditions.

  12. Molecular Characterization of Pathogenic Acanthamoeba Isolated from Drinking and Recreational water in East Azerbaijan, Northwest Iran

    PubMed Central

    Behniafar, Hamed; Niyyati, Maryam; Lasjerdi, Zohreh

    2015-01-01

    Acanthamoeba- related infections, such as amoebic keratitis and granulomatous amoebic encephalitis, can develop in high-risk population through contaminated water sources. Thus, surveying water resources, particularly those available for human use, is of the utmost importance. In the present study, 67 water samples were collected from water resources in East Azerbaijan, a province in northwestern Iran. Samples were cultured on enriched non-nutrient agar plates, and sequencing-based approaches were used for genotyping. The pathogenic potential of the isolates was determined using thermo- and osmo-tolerance tests. Acanthamoeba were detected in 17 (25.4%) of the 67 collected samples. Sequencing analysis revealed that the isolates belonged to the T3 (23.52%), mixed T3/T4 (5.88%), T4 (58.82%), T5 (5.88%), and T13 (5.88%) genotypes. Through thermo- and osmo-tolerance tests, 88.23% of isolates were resistant to 37 °C, 40 °C temperature, and 0.5 M and 1 M osmolarity; thus, these isolates had the potential for pathogenicity. These findings point toa serious public health concern in the studied region. This study is the first to report Acanthamoeba isolated from drinking and recreational water sources in East Azerbaijan and Acanthamoeba T13 isolated from tap water in Iran. PMID:26157334

  13. Fatal Granulomatous Amoebic Encephalitis Caused by Acanthamoeba in a Patient With Kidney Transplant: A Case Report

    PubMed Central

    Salameh, Ahmad; Bello, Nancy; Becker, Jennifer; Zangeneh, Tirdad

    2015-01-01

    Granulomatous amoebic encephalitis (GAE) due to Acanthamoeba is almost a uniformly fatal infection in immune-compromised hosts despite multidrug combination therapy. We report a case of GAE in a female who received a deceased donor kidney graft. She was treated with a combination of miltefosine, pentamidine, sulfadiazine, fluconazole, flucytosine, and azithromycin. PMID:26280011

  14. Essential Role for an M17 Leucine Aminopeptidase in Encystation of Acanthamoeba castellanii.

    PubMed

    Lee, Yu-Ran; Na, Byoung-Kuk; Moon, Eun-Kyung; Song, Su-Min; Joo, So-Young; Kong, Hyun-Hee; Goo, Youn-Kyoung; Chung, Dong-Il; Hong, Yeonchul

    2015-01-01

    Encystation of Acanthamoeba leads to the formation of resilient cysts from vegetative trophozoites. This process is essential for parasite survival under unfavorable conditions such as starvation, low temperatures, and exposure to biocides. During encystation, a massive turnover of intracellular components occurs, and a large number of organelles and proteins are degraded by proteases. Previous studies with specific protease inhibitors have shown that cysteine and serine proteases are involved in encystation of Acanthamoeba, but little is known about the role of metalloproteases in this process. Here, we have biochemically characterized an M17 leucine aminopeptidase of Acanthamoeba castellanii (AcLAP) and analyzed its functional involvement in encystation of the parasite. Recombinant AcLAP shared biochemical properties such as optimal pH, requirement of divalent metal ions for activity, substrate specificity for Leu, and inhibition profile by aminopeptidase inhibitors and metal chelators with other characterized M17 family LAPs. AcLAP was highly expressed at a late stage of encystation and mainly localized in the cytoplasm of A. castellanii. Knockdown of AcLAP using small interfering RNA induced a decrease of LAP activity during encystation, a reduction of mature cyst formation, and the formation of abnormal cyst walls. In summary, these results indicate that AcLAP is a typical M17 family enzyme that plays an essential role during encystation of Acanthamoeba.

  15. Human IgA inhibits adherence of Acanthamoeba polyphaga to epithelial cells and contact lenses.

    PubMed

    Campos-Rodríguez, Rafael; Oliver-Aguillón, Gabriela; Vega-Pérez, Luz M; Jarillo-Luna, Adriana; Hernández-Martínez, Dolores; Rojas-Hernández, Saúl; Rodríguez-Monroy, Marco A; Rivera-Aguilar, Víctor; González-Robles, Arturo

    2004-09-01

    Specific anti-Acanthamoeba IgA antibodies have been detected in the serum and tears of patients and healthy individuals. However, the role of human secretory IgA antibodies in inhibiting the adherence of Acanthamoeba had not been previously investigated. Therefore, the purpose of this study was to purify secretory IgA from human colostrum and analyze its effect on the adherence of Acanthamoeba trophozoites to contact lenses and Madin-Darby canine kidney (MDCK) cells. IgA antibodies to Acanthamoeba polyphaga in colostrum of healthy women as well as in saliva and serum of healthy subjects were analyzed by ELISA and Western blot analysis. In serum, saliva, and colostrum, we detected IgA antibodies that recognized several antigens of A. polyphaga. In addition, colostrum and IgA antibodies purified from it inhibited adherence of A. polyphaga trophozoites to contact lenses and MDCK cells. These results suggest that IgA antibodies may participate in the resistance to the amoebic infection, probably by inhibiting the adherence of the trophozoites to contact lenses and corneal epithelial cells.

  16. Temperature-dependent parasitic relationship between Legionella pneumophila and a free-living amoeba (Acanthamoeba castellanii).

    PubMed

    Ohno, Akira; Kato, Naoyuki; Sakamoto, Ryota; Kimura, Soichiro; Yamaguchi, Keizo

    2008-07-01

    We analyzed the effects of temperature on the interaction of Legionella pneumophila with Acanthamoeba castellanii. At <20 degrees C, overexpression of type 1 metacaspase, a stimulator of A. castellanii encystation, was associated with a reduced number of bacteria within amoeba. At low temperatures, A. castellanii seems to eliminate L. pneumophila by encystation and digestion.

  17. Air bubble in anterior chamber as indicator of full-thickness incisions in femtosecond-assisted astigmatic keratotomy.

    PubMed

    Vaddavalli, Pravin K; Hurmeric, Volkan; Yoo, Sonia H

    2011-09-01

    Femtosecond-assisted astigmatic keratotomy is predictable and precise but may occasionally lead to a full-thickness incision on the cornea and the attendant complications. The presence of an air bubble in the anterior chamber soon after creation of the keratotomy by the femtosecond laser may indicate a full-thickness incision. We present a case in which recognition of this clinical finding early in the procedure might have prevented undesirable complications, such as leakage of aqueous and the potential for intraocular infection.

  18. Propagation of partially coherent twisted anisotropic Gaussian Schell-model beams through an apertured astigmatic optical system.

    PubMed

    Cai, Yangjian; Hu, Li

    2006-03-15

    By expanding the hard-aperture function into a finite sum of complex Gaussian functions, we derived an approximate analytical formula for a partially coherent twisted anisotropic Gaussian Schell-model (AGSM) beam propagating through an apertured paraxial general astigmatic (GA) optical system by use of a tensor method. The results obtained by using the approximate analytical formula are in good agreement with those obtained by using the numerical integral calculation. Our formulas avoid time-consuming numerical integration and provide a convenient and effective way for studying the propagation and transformation of a partially coherent twisted AGSM beam through an apertured paraxial GA optical system.

  19. Interactions between Human Norovirus Surrogates and Acanthamoeba spp.

    PubMed Central

    Hsueh, Tun-Yun

    2015-01-01

    Human noroviruses (HuNoVs) are the most common cause of food-borne disease outbreaks, as well as virus-related waterborne disease outbreaks in the United States. Here, we hypothesize that common free-living amoebae (FLA)—ubiquitous in the environment, known to interact with pathogens, and frequently isolated from water and fresh produce—could potentially act as reservoirs of HuNoV and facilitate the environmental transmission of HuNoVs. To investigate FLA as reservoirs for HuNoV, the interactions between two Acanthamoeba species, A. castellanii and A. polyphaga, as well as two HuNoV surrogates, murine norovirus type 1 (MNV-1) and feline calicivirus (FCV), were evaluated. The results showed that after 1 h of amoeba-virus incubation at 25°C, 490 and 337 PFU of MNV-1/ml were recovered from A. castellanii and A. polyphaga, respectively, while only few or no FCVs were detected. In addition, prolonged interaction of MNV-1 with amoebae was investigated for a period of 8 days, and MNV-1 was demonstrated to remain stable at around 200 PFU/ml from day 2 to day 8 after virus inoculation in A. castellanii. Moreover, after a complete amoeba life cycle (i.e., encystment and excystment), infectious viruses could still be detected. To determine the location of virus associated with amoebae, immunofluorescence experiments were performed and showed MNV-1 transitioning from the amoeba surface to inside the amoeba over a 24-h period. These results are significant to the understanding of how HuNoVs may interact with other microorganisms in the environment in order to aid in its persistence and survival, as well as potential transmission in water and to vulnerable food products such as fresh produce. PMID:25841006

  20. Genetic characterization of clinical acanthamoeba isolates from Japan using nuclear and mitochondrial small subunit ribosomal RNA.

    PubMed

    Rahman, Md Moshiur; Yagita, Kenji; Kobayashi, Akira; Oikawa, Yosaburo; Hussein, Amjad I A; Matsumura, Takahiro; Tokoro, Masaharu

    2013-08-01

    Because of an increased number of Acanthamoeba keratitis (AK) along with associated disease burdens, medical professionals have become more aware of this pathogen in recent years. In this study, by analyzing both the nuclear 18S small subunit ribosomal RNA (18S rRNA) and mitochondrial 16S rRNA gene loci, 27 clinical Acanthamoeba strains that caused AK in Japan were classified into 3 genotypes, T3 (3 strains), T4 (23 strains), and T5 (one strain). Most haplotypes were identical to the reference haplotypes reported from all over the world, and thus no specificity of the haplotype distribution in Japan was found. The T4 sub-genotype analysis using the 16S rRNA gene locus also revealed a clear sub-conformation within the T4 cluster, and lead to the recognition of a new sub-genotype T4i, in addition to the previously reported sub-genotypes T4a-T4h. Furthermore, 9 out of 23 strains in the T4 genotype were identified to a specific haplotype (AF479533), which seems to be a causal haplotype of AK. While heterozygous nuclear haplotypes were observed from 2 strains, the mitochondrial haplotypes were homozygous as T4 genotype in the both strains, and suggested a possibility of nuclear hybridization (mating reproduction) between different strains in Acanthamoeba. The nuclear 18S rRNA gene and mitochondrial 16S rRNA gene loci of Acanthamoeba spp. possess different unique characteristics usable for the genotyping analyses, and those specific features could contribute to the establishment of molecular taxonomy for the species complex of Acanthamoeba.

  1. Prevalence and associations of anisometropia and aniso‐astigmatism in a population based sample of 6 year old children

    PubMed Central

    Huynh, S C; Wang, X Y; Ip, J; Robaei, D; Kifley, A; Rose, K A; Mitchell, P

    2006-01-01

    Aim To study the distribution of anisometropia and aniso‐astigmatism in young Australian children, together with clinical and ocular biometry relations. Method The Sydney Myopia Study examined 1765 predominantly 6 year old children from 34 randomly selected Sydney schools during 2003–4. Keratometry, cycloplegic autorefraction, and questionnaire data were collected. Results Spherical equivalent (SE) anisometropia (⩾1 dioptre) prevalence was 1.6% (95% confidence interval (CI) 1.1% to 2.4%). Aniso‐astigmatism (⩾1D) prevalence was 1.0% (CI: 0.6% to 1.6%). Both conditions were significantly more prevalent among moderately hyperopic (SE ⩾2.0D) than mildly hyperopic (SE 0.5–1.9D) children. Myopic children (SE ⩽−0.5D) had higher anisometropia prevalence. Neither condition varied by age, sex, or ethnicity. In multivariate analyses, anisometropia was significantly associated with amblyopia, odds ratio (OR) 29, (CI: 8.7 to 99), exotropia (OR 7.7, CI: 1.2 to 50), and neonatal intensive care unit (NICU) admission (OR 3.6, CI: 1.1 to 12.6). Aniso‐astigmatism was significantly associated with amblyopia (OR 8.2, CI: 1.4 to 47), maternal age >35 years (OR 4.0, CI: 1.3 to 11.9), and NICU admission (OR 4.6, CI: 1.2 to 17.2). Anisometropia resulted from relatively large interocular differences in axial length (p<0.0001) and anterior chamber depth (p = 0.0009). Aniso‐astigmatism resulted from differences in corneal astigmatism (p<0.0001). Conclusion In this predominantly 6 year old population, anisometropia and aniso‐astigmatism were uncommon, had important birth and biometry associations, and were strongly related to amblyopia and strabismus. PMID:16622090

  2. Necrotizing Meningoencephalitis in a Captive Black and White Ruffed Lemur (Varecia variegata variegata) Caused by Acanthamoeba T4 Genotype.

    PubMed

    Gaide, N; Pelandakis, M; Robveille, C; Albaric, O; Jouvion, G; Souchon, M; Risler, A; Abadie, J

    2015-11-01

    A mature male, black and white ruffed lemur (Varecia variegata variegata) died in a zoological garden after a 4-day history of lethargy and non-responsive convulsions. Necropsy and histopathological examinations revealed acute necrotizing and haemorrhagic meningoencephalitis with intralesional amoebas confirmed by immunohistochemistry. Acanthamoeba T4 genotype was identified as the causative agent of the brain lesion, based on amplification and sequencing of 18S ribosomal RNA genes. The presence of free-living amoebas in water and mud from the lemur's environment was investigated by morphological and molecular analyses. The two predominant genera, representing 80% of isolated amoebas, were Naegleria spp. and Acanthamoeba spp. All Acanthamoeba isolates belonged to the T4 genotype. To the author's knowledge, this is the first report of a meningoencephalitis due to Acanthamoeba T4 genotype in Lemuridae with concurrent analysis of pathological tissues and environment.

  3. Acanthamoeba genotypes T2, T4, and T11 in soil sources from El Hierro island, Canary Islands, Spain.

    PubMed

    Reyes-Batlle, María; Zamora-Herrera, Jonadab; Vargas-Mesa, Alejandro; Valerón-Tejera, Marco Antonio; Wagner, Carolina; Martín-Navarro, Carmen Ma; López-Arencibia, Atteneri; Sifaoui, Ines; Martínez-Carretero, Enrique; Valladares, Basilio; Piñero, José E; Lorenzo-Morales, Jacob

    2016-08-01

    The genus Acanthamoeba includes pathogenic strains which are causative agents of keratitis and encephalitis that often may end fatal in humans and other animals. In the present study, forty soil samples were collected in the island of El Hierro, Canary Islands, Spain, and checked for the presence of Acanthamoeba. Samples were cultivated onto 2 % non-nutrient agar plates seeded with a layer of heat killed Escherichia coli. Amplification by PCR and sequencing of the DF3 region of the 18S rDNA of Acanthamoeba was carried out in order to confirm morphological identification of the amoebae. Furthermore, Acanthamoeba spp. was isolated from 47.5 % of soil samples. Moreover, genotypes T2, T4, and T11 were identified in these samples. To the best of our knowledge, this is the first study to establish genotypes T2, T4, and T11 in soil sources from El Hierro island.

  4. Status of free-living amoebae (Acanthamoeba spp., Naegleria fowleri, Balamuthia mandrillaris) in drinking water supplies in Karachi, Pakistan.

    PubMed

    Yousuf, Farzana Abubakar; Siddiqui, Ruqaiyyah; Subhani, Faysal; Khan, Naveed Ahmed

    2013-06-01

    The ability of pathogenic free-living amoebae to produce infections is a growing concern. In this study, we investigated the presence of free-living amoebae (Acanthamoeba spp., Naegleria fowleri, Balamuthia mandrillaris) in drinking water supplies in Karachi, Pakistan. Fifty-two domestic tap water samples were examined. Amoebae were identified by morphological characteristics and polymerase chain reaction. Thirty percent of the examined samples were positive for Acanthamoeba spp., 8% for N. fowleri while B. mandrillaris were not recovered. Additionally we examined secretory IgA antibody to Acanthamoeba and B. mandrillaris. Acanthamoeba antibody prevalence rate was 100% in both males and females, while B. mandrillaris antibody prevalence rate was 5.5% in males only (females were negative). Our findings suggest that free-living amoebae are a potential health hazard in domestic water supplies in Karachi, Pakistan.

  5. Hypoxia attenuates inflammatory mediators production induced by Acanthamoeba via Toll-like receptor 4 signaling in human corneal epithelial cells

    SciTech Connect

    Pan, Hong; Wu, Xinyi

    2012-04-13

    Highlights: Black-Right-Pointing-Pointer Hypoxia attenuates Acanthamoeba-induced the production of IL-8 and IFN-{beta}. Black-Right-Pointing-Pointer Hypoxia inhibits TLR4 expression in a time-dependent manner in HCECs. Black-Right-Pointing-Pointer Hypoxia inhibits Acanthamoeba-induced the activation of NF-{kappa}B and ERK1/2 in HCECs. Black-Right-Pointing-Pointer Hypoxia decreases Acanthamoeba-induced inflammatory response via TLR4 signaling. Black-Right-Pointing-Pointer LPS-induced the secretion of IL-6 and IL-8 is abated by hypoxia via TLR4 signaling. -- Abstract: Acanthamoeba keratitis (AK) is a vision-threatening corneal infection that is intimately associated with contact lens use which leads to hypoxic conditions on the corneal surface. However, the effect of hypoxia on the Acanthamoeba-induced host inflammatory response of corneal epithelial cells has not been studied. In the present study, we investigated the effect of hypoxia on the Acanthamoeba-induced production of inflammatory mediators interleukin-8 (IL-8) and interferon-{beta} (IFN-{beta}) in human corneal epithelial cells and then evaluated its effects on the Toll-like receptor 4 (TLR4) signaling, including TLR4 and myeloid differentiation primary response gene (88) (MyD88) expression as well as the activation of nuclear factor kappa-light-chain-enhancer of activated B cells (NF-{kappa}B) and extracellular signal-regulated kinases 1/2 (ERK1/2). We then studied the effect of hypoxia on a TLR4-specific inflammatory response triggered by the TLR4 ligand lipopolysaccharide (LPS). Our data showed that hypoxia significantly decreased the production of IL-8 and IFN-{beta}. Furthermore, hypoxia attenuated Acanthamoeba-triggered TLR4 expression as well as the activation of NF-{kappa}B and ERK1/2, indicating that hypoxia abated Acanthamoeba-induced inflammatory responses by affecting TLR4 signaling. Hypoxia also inhibited LPS-induced IL-6 and IL-8 secretion, myeloid differentiation primary response gene (88

  6. Real-time PCR method for the detection and quantification of Acanthamoeba species in various types of water samples.

    PubMed

    Kao, Po-Min; Tung, Min-Che; Hsu, Bing-Mu; Tsai, Hsien-Lung; She, Cheng-Yu; Shen, Shu-Min; Huang, Wen-Chien

    2013-03-01

    In this study, a quantitative real-time PCR was developed to detect and quantify Acanthamoeba spp. in various environmental water samples. The water samples were taken from watershed, water treatment plant, and three thermal spring recreation areas. The overall detection rate was 14.2 % (25/176) for Acanthamoeba spp. The percentages of samples containing Acanthamoeba spp. from river water, raw drinking water, and thermal spring water were 13 % (13/100), 25 % (7/28), and 10.4 % (5/48), respectively. Acanthamoeba spp. concentrations were determined according to SYBR Green quantitative real-time PCR. A plasmid-based standard curve was constructed to determine the Acanthamoeba concentration using dilution factors for achieving 1.36 × 10(9) gene copies per PCR for 18S rRNA gene in Acanthamoeba spp. The resulting concentrations varied by the type of water, in the range of 46-2.6 × 10(2) cells/l in positive raw drinking water, 2.7 × 10(2)-1.5 × 10(4) cells/l in river water, and 54-1.7 × 10(3) cells/l in thermal spring water. The presence of Acanthamoeba spp. in the raw drinking water samples was also found to have a significant difference with heterotrophic plate count. The presence of Acanthamoeba spp. in various aquatic environments may be a potential health hazard and must be further evaluated.

  7. Comparative assessment of astigmatism-corrected Czerny-Turner imaging spectrometer using off-the-shelf optics

    NASA Astrophysics Data System (ADS)

    Yuan, Qun; Zhu, Dan; Chen, Yueyang; Guo, Zhenyan; Zuo, Chao; Gao, Zhishan

    2017-04-01

    We present the optical design of a Czerny-Turner imaging spectrometer for which astigmatism is corrected using off-the-shelf optics resulting in spectral resolution of 0.1 nm. The classic Czerny-Turner imaging spectrometer, consisting of a plane grating, two spherical mirrors, and a sensor with 10-μm pixels, was used as the benchmark. We comparatively assessed three configurations of the spectrometer that corrected astigmatism with divergent illumination of the grating, by adding a cylindrical lens, or by adding a cylindrical mirror. When configured with the added cylindrical lens, the imaging spectrometer with a point field of view (FOV) and a linear sensor achieved diffraction-limited performance over a broadband width of 400 nm centered at 800 nm, while the maximum allowable bandwidth was only 200 nm for the other two configurations. When configured with the added cylindrical mirror, the imaging spectrometer with a one-dimensional field of view (1D FOV) and an area sensor showed its superiority on imaging quality, spectral nonlinearity, as well as keystone over 100 nm bandwidth and 10 mm spatial extent along the entrance slit.

  8. Laser-assisted In Situ Keratomileusis for Correction of Astigmatism and Increasing Contact Lens Tolerance after Penetrating Keratoplasty

    PubMed Central

    Park, Chang-Hyun; Kim, Su-Young

    2014-01-01

    Purpose To determine effectiveness of laser-assisted in situ keratomileusis (LASIK) in the treatment of astigmatism following penetrating keratoplasty (PK). Methods We performed a retrospective review of medical records of patients who underwent LASIK following PK and had over 1 year of follow-up data. Results Twenty-six patients (26 pairs of eyes) underwent LASIK following PK. Mean age of the patients at the time of LASIK was 40.7 years (range, 26 to 72 years). Following LASIK, the mean cylinder was reduced by 2.4 diopters and mean reduction of cylinder after LASIK was 65.4% from the preoperative values at the last follow-up visit. Uncorrected visual acuity became 20 / 50 or better in 69.2% of the eyes after LASIK. Best-corrected visual acuity became 20 / 50 or better in 73.1% of the eyes after LASIK. All of them were intolerable to contact lenses before LASIK. After LASIK, 6 pairs (23.1%) did not need to use contact lenses and 18 pairs (69.2%) were tolerable to using contact lenses or spectacles. There were no significant endothelial cell density changes 12 months after LASIK (p = 0.239). Conclusions LASIK is effective in the treatment of astigmatism following PK and increases contact lens and spectacle tolerance. PMID:25276076

  9. Computation of astigmatic and trefoil figure errors and misalignments for two-mirror telescopes using nodal-aberration theory.

    PubMed

    Ju, Guohao; Yan, Changxiang; Gu, Zhiyuan; Ma, Hongcai

    2016-05-01

    In active optics systems, one concern is how to quantitatively separate the effects of astigmatic and trefoil figure errors and misalignments that couple together in determining the total aberration fields when wavefront measurements are available at only a few field points. In this paper, we first quantitatively describe the impact of mount-induced trefoil deformation on the net aberration fields by proposing a modified theoretical formulation for the field-dependent aberration behavior of freeform surfaces based on the framework of nodal aberration theory. This formulation explicitly expresses the quantitative relationships between the magnitude of freeform surfaces and the induced aberration components where the freeform surfaces can be located away from the aperture stop and decentered from the optical axis. On this basis, and in combination with the mathematical presentation of nodal aberration theory for the effects of misalignments, we present the analytic expressions for the aberration fields of two-mirror telescopes in the presence of astigmatic primary mirror figure errors, mount-induced trefoil deformations on both mirrors, and misalignments. We quantitatively separate these effects using the analytical expressions with wavefront measurements at a few field points and pointing errors. Valuable insights are provided on how to separate these coupled effects in the computation process. Monte Carlo simulations are conducted to demonstrate the correctness and accuracy of the analytic method presented in this paper.

  10. Fulminant and fatal encephalitis caused by Acanthamoeba in a kidney transplant recipient: case report and literature review.

    PubMed

    Satlin, M J; Graham, J K; Visvesvara, G S; Mena, H; Marks, K M; Saal, S D; Soave, R

    2013-12-01

    Acanthamoeba is the most common cause of granulomatous amebic encephalitis, a typically fatal condition that is classically described as indolent and slowly progressive. We report a case of Acanthamoeba encephalitis in a kidney transplant recipient that progressed to death within 3 days of symptom onset and was diagnosed at autopsy. We also review clinical characteristics, treatments, and outcomes of all published cases of Acanthamoeba encephalitis in solid organ transplant (SOT) recipients. Ten cases were identified, and the infection was fatal in 9 of these cases. In 6 patients, Acanthamoeba presented in a fulminant manner and death occurred within 2 weeks after the onset of neurologic symptoms. These acute presentations are likely related to immunodeficiencies associated with solid organ transplantation that result in an inability to control Acanthamoeba proliferation. Skin lesions may predate neurologic involvement and provide an opportunity for early diagnosis and treatment. Acanthamoeba is an under-recognized cause of encephalitis in SOT recipients and often presents in a fulminant manner in this population. Increased awareness of this disease and its clinical manifestations is essential to attain an early diagnosis and provide the best chance of cure.

  11. Isolation and molecular characterization of Acanthamoeba genotypes isolated from soil sources of public and recreational areas in Iran.

    PubMed

    Karamati, Seyed Ahmad; Niyyati, Maryam; Lorenzo-Morales, Jacob; Lasjerdi, Zohreh

    2016-12-01

    Pathogenic strains of Acanthamoeba are causative agents of a sight threating infection of the cornea known as Acanthamoeba keratitis. AK cases have been reported in Iran recently due to inappropriate usage of contact lens maintenance and most patients report a contact with contaminated sources such as dust, water or soil. Sixty soil samples were collected from public and recreational areas in the province of East Azerbaijan, Iran and checked for the presence of Acanthamoeba spp. Samples were cultured on non-nutrient agar plates seeded with heat killed Escherichia coli. PCR and sequencing of the DF3 region were carried out in order to genotype the isolated strains of Acanthamoeba. Thermotolerance and osmotolerance assays were performed in order to investigate the pathogenic potential of isolated Acanthamoeba strains. Acanthamoeba spp. was isolated from 41.6% of soil samples and genotyping of the strains resulted in the identification of genotypes T3, T4, T5 and T11. Most of the isolates belonging to genotypes T3 and T4 showed high pathogenic potential, indicating that they might present a potential health hazard for humans and other animals in this region. To the best of our knowledge, this is the first report on the identification of genotypes T3 and T11 from soil sources in the country.

  12. The first genotype determination of Acanthamoeba potential threat to human health, isolated from natural water reservoirs in Poland.

    PubMed

    Lass, Anna; Szostakowska, Beata; Idzińska, Alicja; Chomicz, Lidia

    2014-07-01

    Different species of amoebae belonging to the genus Acanthamoeba are widely distributed in many parts of the world and known as free-living organisms. Some strains of the protozoans may exist as parasites and cause risk to human health as causative agents of serious human diseases. Currently, in Poland, there is no sufficient information about the distribution of Acanthamoeba strains and their genotypes in the environment. Therefore, 20 environmental surface water samples were collected from different sites located at five water reservoirs in Gdynia, Sopot, and Gdańsk (northern Poland). The material was cultured to obtain Acanthamoeba isolates that were then specifically analyzed with both PCR and real-time PCR assays. Of the 20 samples examined, Acanthamoeba DNA was found in 13 samples tested with the use of real-time PCR; in 10 of them, DNA of the amoeba was also detected using PCR technique. The comparison with sequences available in the GenBank confirmed that the PCR products are fragments of Acanthamoeba 18S rRNA gene and that isolates represent T4 genotype, known as the most common strains related to AK cases. This is the first investigation in Poland describing Acanthamoeba detection in environmental water samples with molecular techniques and genotyping. The results indicate that surface water in Poland may be a source of acanthamoebic strains potentially pathogenic for humans.

  13. Isolation and genotyping of free-living environmental isolates of Acanthamoeba spp. from bromeliads in Southern Brazil.

    PubMed

    Landell, Melissa Fontes; Salton, Juliana; Caumo, Karin; Broetto, Leonardo; Rott, Marilise B

    2013-07-01

    Species of Acanthamoeba are frequently isolated from distinct environmental sources such as water, soil, dust and air. They are responsible to cause infections and disease in humans and animals. In addition, Acanthamoeba sp. are considered an important reservoir of bacteria, virus and fungi, which act as "Trojan horses" to protect these microorganisms of harsh environmental conditions. In this study, nine Acanthamoeba isolates from bromeliads phylloplane were identified based on the morphology of cyst and trophozoite forms. The genotype level was accessed by the sequence analysis of Acanthamoeba small-subunit rRNA gene. Genotypic characterization grouped five isolates in the genotype T2/T6, three in the T4 genotype and one in the genotype T16. The results obtained indicate that the genotype T2/T6 is common on phylloplane. To predict the pathogenic potential of the Acanthamoeba isolates, thermo and osmotolerance assays were employed, although all isolates were capable of surviving at temperatures of 37°C, other tests will be conducted in the future to determine the potential pathogenic of the isolates. Altogether, our results revealed the importance of the presence of Acanthamoeba associated with bromeliads in Rio Grande do Sul, Brazil, and the necessity for further studies to determine the environmental distribution and the role of these species.

  14. Acanthamoeba infection in lungs of mice expressed by toll-like receptors (TLR2 and TLR4).

    PubMed

    Derda, Monika; Wojtkowiak-Giera, Agnieszka; Kolasa-Wołosiuk, Agnieszka; Kosik-Bogacka, Danuta; Hadaś, Edward; Jagodziński, Paweł P; Wandurska-Nowak, Elżbieta

    2016-06-01

    Toll-like receptors (TLRs) play a key role in the innate immune responses to a variety of pathogens including parasites. TLRs are among the most highly conserved in the evolution of the receptor family, localized mainly on cells of the immune system and on other cells such as lung cells. The aim of this study was to determine for the first time the expression of TLR2 and TLR4 in the lung of Acanthamoeba spp. infected mice using quantitative real-time polymerase chain reaction (Q-PCR) and immunohistochemical (IHC) staining. The Acanthamoeba spp. were isolated from a patient with Acanthamoeba keratitis (AK) (strain Ac 55) and from environmental samples of water from Malta Lake (Poznań, Poland - strain Ac 43). We observed a significantly increased level of expression of TLR2 as well as TLR4 mRNA from 2 to 30 days post Acanthamoeba infection (dpi) in the lungs of mice infected with Ac55 (KP120880) and Ac43 (KP120879) strains. According to our observations, increased TLR2 and TLR4 expression in the pneumocytes, interstitial cells and epithelial cells of the bronchial tree may suggest an important role of these receptors in protective immunity against Acanthamoeba infection in the lung. Moreover, increased levels of TLR2 and TLR4 mRNA expression in infected Acanthamoeba mice may suggest the involvement of these TLRs in the recognition of this amoeba pathogen-associated molecular pattern (PAMP).

  15. Isolation and molecular characterization of Acanthamoeba genotypes in recreational and domestic water sources from Jamaica, West Indies.

    PubMed

    Todd, Cheridah D; Reyes-Batlle, María; Piñero, José E; Martínez-Carretero, Enrique; Valladares, Basilio; Streete, Don; Lorenzo-Morales, Jacob; Lindo, John F

    2015-09-01

    Free living amoebae (FLA) are amphizoic protozoa that are ubiquitous in nature. Infection with FLA may result in neurological, ocular and skin infections. Exposure to Acanthamoeba occurs frequently through water contact and knowledge of the presence of the organisms in water sources is important in understanding transmission dynamics. The distribution of Acanthamoeba was studied in recreational and domestic water samples collected from across Jamaica. Morphological assessment and polymerase chain reaction revealed Acanthamoeba spp. isolates in 50.6% (42/83) and 17.3% (14/81) of recreational and domestic water, respectively. Sequencing of the DF3 region of the 18S rDNA resulted in the identification of genotypes T3, T4, T5, T10 and T11 corresponding to Acanthamoeba spp: A. griffini, A. triangularis, A. lenticulata, A. culbertsoni and A. hatchetti. Moreover, T4 was the most frequently isolated genotype in both recreational and domestic water. Thermotolerance and osmotolerance assays indicated that most isolates were potentially pathogenic. This is the first report of T3 and T10 genotypes in the Caribbean and the first report of these Acanthamoeba spp. in Jamaican waters. The study shows that there is potential risk of infection to contact wearers who practise poor lens care. Further, Acanthamoeba should be considered as a cause of neurological infections in Jamaica.

  16. Immunological inter-strain crossreactivity correlated to 18S rDNA sequence types in Acanthamoeba spp.

    PubMed

    Walochnik, J; Obwaller, A; Aspöck, H

    2001-02-01

    Various species of the genus Acanthamoeba have been described as potential pathogens; however, differentiation of acanthamoebae remains problematic. The genus has been divided into 12 18S rDNA sequence types, most keratitis causing strains exhibiting sequence type T4. We recently isolated a keratitis causing Acanthamoeba strain showing sequence type T6, but being morphologically identical to a T4 strain. The aim of our study was to find out, whether the 18S rDNA sequence based identification correlates to immunological differentiation. The protein and antigen profiles of the T6 isolate and three reference Acanthamoeba strains were investigated using two sera from Acanthamoeba keratitis patients and one serum from an asymptomatic individual. It was shown, that the T6 strain produces a distinctly different immunological pattern, while patterns within T4 were identical. Affinity purified antibodies were used to further explore immunological cross-reactivity between sequence types. Altogether, the results of our study support the Acanthamoeba 18S rDNA sequence type classification in the investigated strains.

  17. Acanthamoeba misidentification and multiple labels: redefining genotypes T16, T19, and T20 and proposal for Acanthamoeba micheli sp. nov. (genotype T19).

    PubMed

    Corsaro, Daniele; Walochnik, Julia; Köhsler, Martina; Rott, Marilise B

    2015-07-01

    Acanthamoeba species are ubiquitous amoebae able to cause important infections in humans and other vertebrates. The full/near-full sequences (>2000 bp) of the small subunit ribosomal RNA gene (SSU rDNA or 18S rDNA) are used to cluster Acanthamoeba as genotypes, labeled T1 to T20. Genotype T15 remains an exception, being described only partially on a 1500-bp fragment. Strains are thus usually identified based on their 18S identity matches with reference strains, often using shorter (<500 bp) diagnostic fragments of the gene. Nevertheless, short fragments (<1000 bp) have been used to propose genotypes. This has been criticized, and doubts arise therefore on possible confusion leading to classify distinct partial sequences with a same label(s). We demonstrate herein that several partial sequences misassigned either to T16 or to T4, actually belong to at least two separate and distinct genotypes. We obtained the full 18S rDNA of a strain previously typed as T16 on the basis of a small fragment and demonstrated that it actually belongs to the recently described T19. We propose the name Acanthamoeba micheli sp. nov., for this strain. Furthermore, partial molecular phylogenies were performed to show that several other misassigned T16 partial sequences belong to a new genotype. This latter includes also misassigned T4 partial sequences, only recently available as full sequences and labeled as T20. We thus reassign these partial sequences to the genotype T20. Longer sequences, ideally at least 90 % of the total gene length, should be obtained from strains to ensure reliable diagnostic and phylogenetic results.

  18. Comparison of Visian toric collamer lens and toric acrylic intraocular lens implantation for the treatment of myopia with astigmatism

    PubMed Central

    Ammar, Hatem; Anbar, Mohamed; Abdellah, Marwa M

    2017-01-01

    Purpose To compare the efficacy and outcome of phakic toric implantable collamer lens (TICL) and refractive clear lens extraction with AcrySof Toric intraocular lens (TIOL) implantation for the treatment of myopic astigmatism. Patients and methods This study assessed eyes with myopic astigmatism >−1 D and ≤−4 D with a spherical equivalent >10 D or <10 D if the patients were unsuitable for corneal refractive surgery. These eyes were divided into group A, in which Visian Toric ICL™ Phakic TICL was implanted, and group B, which involved clear lens extraction with implantation of an AcrySof IQ toric SN60T3-9™ IOL. The outcome and complications were evaluated. Results This study enrolled 63 eyes of 38 patients with a follow-up period of at least 6 months. The mean postoperative spherical equivalent was −0.19±0.31 D in group A and −0.21±0.28 D in group B (P=0.69). The mean postoperative cylinder value was −0.46±0.53 D in group A and −0.32±0.41 D in group B (P=0.35). Postoperative cylinder was <1 D in 76.47% and 79.31% of eyes in groups A and B, respectively. The mean endothelial cell count was reduced by 4.32% in group A and by 5.32% in group B (P=0.003). The mean postoperative intraocular pressure increased insignificantly in group A (P=0.22) and reduced significantly in group B (P=0.004). The complication rate was 11.76% in group A and 6.90% in group B. Conclusion Both procedures showed predictable results and good visual results. However, the loss of accommodation and risk of retinal complications in the TIOL group suggest that the use of TICL for myopic astigmatism is a better choice in younger patients. PMID:28096654

  19. Distribution of Acanthamoeba Genotypes Isolated from Recreational and Therapeutic Geothermal Water Sources in Southwestern Iran.

    PubMed

    Niyyati, Maryam; Saberi, Reza; Latifi, Alireza; Lasjerdi, Zohreh

    2016-01-01

    A comprehensive survey was conducted along 10 km of geothermal rivers in southwestern Iran. A total of 40 water samples were tested for the presence of Acanthamoeba spp., and genotypes were determined by targeting the diagnostic fragment 3 region of the 18S rRNA gene. The pathogenic potential of all positive isolates was also identified using tolerance ability test. High occurrences of Acanthamoeba (50%) were detected in the sampling areas. Based on sequencing analysis, isolates belonging to T4 (93.7%) and T2 (6.25%) genotypes were reported. Thermo- and osmotolerance tests revealed that five strains are highly pathogenic. Since every collection site of this study was associated with high human activity, posting of warning signs, monitoring of recreational water sources, and awareness of high-risk people are of utmost importance. To the best of our knowledge, the present research is the first to report T2 genotype from geothermal water sources in Iran.

  20. Comparison of Fluorescence Microscopy and Different Growth Media Culture Methods for Acanthamoeba Keratitis Diagnosis

    PubMed Central

    Peretz, Avi; Geffen, Yuval; Socea, Soergiu D.; Pastukh, Nina; Graffi, Shmuel

    2015-01-01

    Acanthamoeba keratitis (AK), a potentially blinding infection of the cornea, is caused by a free-living protozoan. Culture and microscopic examination of corneal scraping tissue material is the conventional method for identifying Acanthamoeba. In this article, we compared several methods for AK diagnosis of 32 patients: microscopic examination using fluorescent dye, specific culture on growth media—non-nutrient agar (NNA), culture on liquid growth media—peptone yeast glucose (PYG), and TYI-S-33. AK was found in 14 patients. Thirteen of the specimens were found AK positive by fluorescence microscopic examination, 11 specimens were found AK positive on PYG growth media, and 9 specimens were found AK positive on TYI-S-33 growth media. Only five specimens were found AK positive on NNA growth media. Therefore, we recommend using fluorescence microscopy technique and culture method, especially PYG liquid media. PMID:25962772

  1. Superdiffusion dominates intracellular particle motion in the supercrowded cytoplasm of pathogenic Acanthamoeba castellanii

    NASA Astrophysics Data System (ADS)

    Reverey, Julia F.; Jeon, Jae-Hyung; Bao, Han; Leippe, Matthias; Metzler, Ralf; Selhuber-Unkel, Christine

    2015-06-01

    Acanthamoebae are free-living protists and human pathogens, whose cellular functions and pathogenicity strongly depend on the transport of intracellular vesicles and granules through the cytosol. Using high-speed live cell imaging in combination with single-particle tracking analysis, we show here that the motion of endogenous intracellular particles in the size range from a few hundred nanometers to several micrometers in Acanthamoeba castellanii is strongly superdiffusive and influenced by cell locomotion, cytoskeletal elements, and myosin II. We demonstrate that cell locomotion significantly contributes to intracellular particle motion, but is clearly not the only origin of superdiffusivity. By analyzing the contribution of microtubules, actin, and myosin II motors we show that myosin II is a major driving force of intracellular motion in A. castellanii. The cytoplasm of A. castellanii is supercrowded with intracellular vesicles and granules, such that significant intracellular motion can only be achieved by actively driven motion, while purely thermally driven diffusion is negligible.

  2. Sequence organization of the Acanthamoeba rRNA intergenic spacer: identification of transcriptional enhancers.

    PubMed Central

    Yang, Q; Zwick, M G; Paule, M R

    1994-01-01

    The primary sequence of the entire 2330 bp intergenic spacer of the A.castellanii ribosomal RNA gene was determined. Repeated sequence elements averaging 140 bp were identified and found to bind a protein required for optimum initiation at the core promoter. These repeated elements were shown to stimulate rRNA transcription by RNA polymerase I in vitro. The repeats inhibited transcription when placed in trans, and stimulated transcription when in cis, in either orientation, but only when upstream of the core promoter. Thus, these repeated elements have characteristics similar to polymerase I enhancers found in higher eukaryotes. The number of rRNA repeats in Acanthamoeba cells was determined to be 24 per haploid genome, the lowest number so far identified in any eukaryote. However, because Acanthamoeba is polyploid, each cell contains approximately 600 rRNA genes. Images PMID:7984432

  3. Assessment of the efficacy of benzalkonium chloride and sodium hypochlorite against Acanthamoeba polyphaga and Tetrahymena spp.

    PubMed

    Vaerewijck, M J M; Sabbe, K; Baré, J; Spengler, H-P; Favoreel, H W; Houf, K

    2012-03-01

    The efficacy of benzalkonium chloride and sodium hypochlorite against Acanthamoeba polyphaga and two Tetrahymena spp. was determined based on the European Standard EN 1276:2009 suspension test. Trophozoite viability was assessed by determination of the membrane integrity using flow cytometry as a fast screening technique. Bovine serum albumin was added to simulate clean (0.3 g/liter) and dirty (3 g/liter) conditions. Benzalkonium chloride caused cell lysis at concentrations above 50 mg/liter under clean and dirty conditions. A concentration of 50 mg of free chlorine per liter had a strong biocidal effect on acanthamoebae and tetrahymenae after 15 min under clean and dirty conditions. Our results suggest that benzalkonium chloride and sodium hypochlorite were effective against the three microorganisms at concentrations commonly applied in the food industry.

  4. Isolation of acanthamoeba genotype t4 from a non-contact lens wearer from the Philippines.

    PubMed

    Buerano, Corazon C; Trinidad, Abigail D; Fajardo, Lindsay Sydney N; Cua, Irwin Y; Baclig, Michael O; Natividad, Filipinas F

    2014-12-01

    We report the case of a 76-year old Filipino male who presented with pain, redness, and blurring of vision of the right eye. Corneal scraping was done and sent to the St. Luke's Research and Biotechnology Group for detection and identification of the infectious agent. Morphological detection was performed by allowing the organism from the scraping to grow in 1.5% non-nutrient agar plate with heat-killed E. coli. Trophozoites with acanthopodia and double-walled cysts characteristic of Acanthamoeba were observed within the first and second week of observations, respectively. Molecular identification of the amoebae at the genus level based on the presence of Acanthamoeba-specific amplimer S1, ASA.S1 confirmed the morphological identification. Genotyping through sequence revealed that the organism belonged to T4, which is the genotype commonly present in the eye of keratitis patients.

  5. Isolation of Acanthamoeba Genotype T4 from a Non-Contact Lens Wearer from the Philippines

    PubMed Central

    Buerano, Corazon C.; Trinidad, Abigail D.; Fajardo, Lindsay Sydney N.; Cua, Irwin Y.; Baclig, Michael O.; Natividad, Filipinas F.

    2014-01-01

    We report the case of a 76-year old Filipino male who presented with pain, redness, and blurring of vision of the right eye. Corneal scraping was done and sent to the St. Luke’s Research and Biotechnology Group for detection and identification of the infectious agent. Morphological detection was performed by allowing the organism from the scraping to grow in 1.5% non-nutrient agar plate with heat-killed E. coli. Trophozoites with acanthopodia and double-walled cysts characteristic of Acanthamoeba were observed within the first and second week of observations, respectively. Molecular identification of the amoebae at the genus level based on the presence of Acanthamoeba-specific amplimer S1, ASA.S1 confirmed the morphological identification. Genotyping through sequence revealed that the organism belonged to T4, which is the genotype commonly present in the eye of keratitis patients. PMID:25589879

  6. Bringing forward the new generation of alkoxy-thiourea as potential treatment for Acanthamoeba keratitis

    NASA Astrophysics Data System (ADS)

    Khairul, Wan M.; Goh, Yit-Peng; Daud, Adibah Izzati; Nakisah, M. A.

    2017-02-01

    Alkoxy substituted thiourea derivatives with general formula of A-ArC(O)NHC(S)NHAr-D which A represents the methoxy group and D denotes -OCnH2n+1 have been successfully synthesised and characterized. In turn, all the synthesised molecules were assayed for anti-amoebic activities towards Acanthamoeba sp to examine the cytotoxicity effect at their IC50 and membrane permeability. As predicted, the findings showed that the synthesised molecules owing promising anti-amoebic activity towards Acanthamoeba sp. To support, the Acridine-orange/Propidium iodide (AOPI) staining result under fluorescence microscopy revealed the treated amoeba cells by these alkoxy thiourea derivatives exhibited loss in their membrane permeability.

  7. Distribution of Acanthamoeba Genotypes Isolated from Recreational and Therapeutic Geothermal Water Sources in Southwestern Iran

    PubMed Central

    Niyyati, Maryam; Saberi, Reza; Latifi, Alireza; Lasjerdi, Zohreh

    2016-01-01

    A comprehensive survey was conducted along 10 km of geothermal rivers in southwestern Iran. A total of 40 water samples were tested for the presence of Acanthamoeba spp., and genotypes were determined by targeting the diagnostic fragment 3 region of the 18S rRNA gene. The pathogenic potential of all positive isolates was also identified using tolerance ability test. High occurrences of Acanthamoeba (50%) were detected in the sampling areas. Based on sequencing analysis, isolates belonging to T4 (93.7%) and T2 (6.25%) genotypes were reported. Thermo- and osmotolerance tests revealed that five strains are highly pathogenic. Since every collection site of this study was associated with high human activity, posting of warning signs, monitoring of recreational water sources, and awareness of high-risk people are of utmost importance. To the best of our knowledge, the present research is the first to report T2 genotype from geothermal water sources in Iran. PMID:27127409

  8. An atypical presentation of Acanthamoeba keratitis in a noncontact lens wearer.

    PubMed

    Speer, Christine E; Hofmeister, Elizabeth M; Cohen, Elisabeth J

    2003-01-01

    This article presents the case of a 49-year-old man who did not have a history of wearing contact lenses and who developed a rapidly progressive course of Acanthamoeba keratitis. The patient developed stromal keratitis that did not respond to herpes simplex virus therapies. Within 1 week after presentation, the patient progressed from mild anterior stromal haze and edema to a ring infiltrate, epithelial loss, and significant corneal edema. Corneal scrapings demonstrated cysts consistent with Acanthanmoeba keratitis. The patient was admitted to the hospital and placed on intensive medical therapy. He responded to therapy, and at 5 months showed central scarring in a quiet eye. This article presents a case of Acanthamoeba keratitis in a non-contact lens wearer, who was diagnosed clinically and histopathologically within 1 week of onset of symptoms. His case was atypical given his lack of contact lens wear or antecedent trauma and rapid progression to a ring infiltrate, usually seen as late findings.

  9. [Acanthamoeba keratitis after use of soft contact lenses--case report].

    PubMed

    Ziak, P; Ondriska, F; Mrva, M

    2003-09-01

    The case history of a 39-year patient suffering from a deep inflammation of cornea and not responding to conventional antibiotic treatment is presented. The patient was using soft contact lenses during the period of initial symptoms; moreover, he was bathing in thermal bathing pool. A cultivation examination of smears from the area of corneal defect revealed the presence of Acanthamoeba lugdunensis in combination with bacterial infection by Pseudomonas aeruginosa. The available data indicate that it is the first case of acanthamoeba karatitis (AK) after the application of contact lenses in Slovakia. A long-term local treatment with propamidin isethionate (Brolene gtt, ung.) resulted in healing up. The subsequent vision after 16 months since the initial symptoms proved to be 6/12 (0.5). The healing of the centrally localized defect changed the curvature of cornea with consequent hypermetropic shift. The defect completely corrected the patient's myopia (-8.5). The paper describes present possibilities of AK therapy.

  10. In Vitro Evaluation of the Effectiveness of the Macrolide Rokitamycin and Chlorpromazine against Acanthamoeba castellanii

    PubMed Central

    Mattana, A.; Biancu, G.; Alberti, L.; Accardo, A.; Delogu, G.; Fiori, P. L.; Cappuccinelli, P.

    2004-01-01

    The present study demonstrates the in vitro effectiveness of the macrolide rokitamycin and the phenothiazine compound chlorpromazine against Acanthamoeba castellanii. Growth curve evaluations revealed that both drugs inhibit trophozoite growth in dose- and time-dependent ways. The effects of both drugs when they were used at the MICs at which 100% of isolates are inhibited were amoebistatic, but at higher doses they were amoebicidal as well as cysticidal. Experiments showed that when rokitamycin was associated with chlorpromazine or amphotericin B, rokitamycin enhanced their activities. Furthermore, low doses of rokitamycin and chlorpromazine, alone or in combination, blocked the cytopathic effect of A. castellanii against WKD cells derived from the human cornea. These results may have important significance in the development of new anti-Acanthamoeba compounds. PMID:15561820

  11. Clinical evaluation of surgery-induced astigmatism in cataract surgery using 2.2 mm or 1.8 mm clear corneal micro-incisions

    PubMed Central

    Yang, Jun; Wang, Xiu; Zhang, Hong; Pang, Yi; Wei, Rui-Hua

    2017-01-01

    AIM To evaluate corneal astigmatism after phacoemulsification using 2.2 mm or 1.8 mm clear corneal micro-incisions and its effects on visual function. METHODS Sixty cases (60 eyes) with cataract were randomly divided into groups A (n=30) and B (n=30) respectively underwent 2.2 mm and 1.8 mm clear corneal tunnel incision phacoemulsification combined with folding intraocular lens implantation from the time direction of 11:00. On day 1 and at 1, 4, and 6wk after operation, patients' vision was measured and both the corneal curvature and corneal thickness (CT) were recorded using Pentacam. RESULTS The measured surgery-induced astigmatism (SIA) in both groups A and B peaked on day 1 after operation, and then gradually decreased and eventually stabilized in week 4. No statistically significant difference was found in corneal astigmatism between two groups (P>0.05). The measured corneal astigmatism at 4wk and 6wk postoperatively were 0.28±0.09 D and 0.27±0.10 D for groups A and 0.27±0.09 D and 0.25±0.10 D for groups B without statistically significant difference (P>0.05). In addition, no significant differences in visual acuity and CT were found between groups A and B before or after operation. CONCLUSION Both 2.2 mm and 1.8 mm micro-incision cataract surgeries result in relatively small SIA with no difference in visual function and corneal astigmatism between two surgery approaches. Thus, the two types of surgical systems are safe and efficient for cataract treatment, by which satisfactory uncorrected visual acuity can be regained early postoperatively. PMID:28149779

  12. Comparative analysis of the efficacy of astigmatic correction after wavefront-guided and wavefront-optimized LASIK in low and moderate myopic eyes

    PubMed Central

    Khalifa, Mounir A.; Alsahn, Mahmoud F.; Shaheen, Mohamed Shafik; Pinero, David P.

    2017-01-01

    AIM To evaluate and compare the efficacy of the astigmatic correction achieved with laser in situ keratomileusis (LASIK) in eyes with myopic astigmatism using wavefront-guided (WFG) and wavefront-optimized (WFO) ablation profiles. METHODS Prospective study included 221 eyes undergoing LASIK: 99 and 122 eyes with low and moderate myopic astigmatism (low and moderate myopia groups). Two subgroups were differentiated in each group according to the ablation profile: WFG subgroup, 109 eyes (45/64, low/moderate myopia groups) treated using the Advanced CustomVue platform (Abbott Medical Optics Inc.), and WFO subgroup, 112 eyes (54/58, low/moderate myopia groups) treated using the EX-500 platform (Alcon). Clinical outcomes were evaluated during a 6-month follow-up, including a vector analysis of astigmatic changes. RESULTS Significantly better postoperative uncorrected visual acuity and efficacy index was found in the WFG subgroups of each group (P≤0.041). Postoperative spherical equivalent and cylinder were significantly higher in WFO subgroups (P≤0.003). In moderate myopia group, a higher percentage of eyes with a postoperative cylinder ≤0.25 D was found in the WFG subgroup (90.6% vs 65.5%, P=0.002). In low and moderate myopia groups, the difference vector was significantly higher in the WFO subgroup compared to WFG (P<0.001). In moderate myopia group, the magnitude (P=0.008) and angle of error (P<0.001) were also significantly higher in the WFO subgroup. Significantly less induction of high order aberrations were found with WFG treatments in both low and moderate myopia groups (P≤0.006). CONCLUSION A more efficacious correction of myopic astigmatism providing a better visual outcome is achieved with WFG LASIK compared to WFO LASIK. PMID:28251090

  13. A Rabbit Model of Acanthamoeba Keratitis That Better Reflects the Natural Human Infection.

    PubMed

    Feng, Xianmin; Zheng, Wenyu; Wang, Yuehua; Zhao, Donghai; Jiang, Xiaoming; Lv, Shijie

    2015-08-01

    Acanthamoeba species are ubiquitous, free-living protozoa that can invade the cornea and result in Acanthamoeba keratitis (AK), a painful progressive sight-threatening corneal disease. Disease progression in current animal models is too rapid to mimic AK in humans accurately. This study provides a novel method for establishing AK in rabbits and compared it with the conventional method with regard to pathogenesis and immune response in humans. The New Zealand white rabbits were randomly divided into two experimental groups (Groups A and B). Rabbits in the Group A (n = 14) received intrastromal injections of 1 × 10(4) /100 µL Acanthamoeba healyi trophozoites (conventional AK model). The Group B animals (n = 14) received microinjections of 1 × 10(4) /10 µL A. healyi trophozoites between the corneal epithelium and Bowman's layer, anterior to the corneal stroma (novel AK model). In addition, two rabbits were left untreated as normal controls. AK in the treated rabbits was evaluated clinically, histopathologically, and immunologically for 35 days. AK was successfully established in both the conventional and novel model groups. Compared with the Group A, AK in the Group B displayed an efficient immune response with less severe pathology. Moreover, the self-limiting but chronic nature of the infection in the Group B was strikingly similar to that of AK in humans. The novel animal model for AK described here more closely simulates the pathogenesis and immune response of Acanthamoeba corneal infection in humans than the animal models currently in use.

  14. Prevalence of Acanthamoeba spp. (Sarcomastigophora: Acanthamoebidae) in wild populations of Aedes aegypti (Diptera: Culicidae).

    PubMed

    Otta, Dayane Andriotti; Rott, Marilise Brittes; Carlesso, Ana Maris; da Silva, Onilda Santos

    2012-11-01

    Studies of interrelationship between microorganisms and mosquitoes are of great importance, since it can provide support for better understand related to biology, development and their control. In this way, it is known that mosquito larvae and free-living amoebae (FLA) normally occupy similar aquatic microhabitats. However, few studies have been conducted about such coexistence. For that reason, the objective of the present study was to verify the prevalence of Acanthamoeba spp. in wild populations of Aedes aegypti, as well as to characterize the genotypic lineage, and their possible pathogenicity through thermo- and osmotolerance. Amoebae were investigated in 60 pools, each containing ten larvae of A. aegypti, collected in Porto Alegre (Rio Grande do Sul, Brazil). The Acanthamoeba isolates were morphologically characterized and submitted to the polymerase chain reaction technique to confirm identification of the genus. In addition, genotype analyses as well as tests for presumptive pathogenicity in some samples were performed. Of the 60 pools examined, 54 (90 %) were positive for FLA. Of these isolates, 47 (87 %) belonged to the genus Acanthamoeba. The genotypic groups T4, T3 and T5 were identified, numbering 14 (53.8 %), ten (38.5 %) and two (7.7 %) isolates, respectively. The physiological tests performed with 14 strains showed that 12 (85.7 %) were non-pathogenic, while two (14.3 %) were considered as having low pathogenic potential. These results provide a basis for a better understanding of the interaction between these protozoan and mosquitoes in their natural habitat. This study is the first to report the isolation of Acanthamoeba spp. from wild mosquitoes.

  15. Experimental infection of T4 Acanthamoeba genotype determines the pathogenic potential.

    PubMed

    Alves, Daniella de Sousa Mendes Moreira; Moraes, Aline Silva; Alves, Luciano Moreira; Gurgel-Gonçalves, Rodrigo; Lino Junior, Ruy de Souza; Cuba-Cuba, César Augusto; Vinaud, Marina Clare

    2016-09-01

    T4 is the Acanthamoeba genotype most related to cases of granulomatous amoebic encephalitis (GAE) in immunocompromised patients and of keratitis in contact lens wearers. The determination of the pathogenic potential of Acanthamoeba clinical and environmental isolates using experimental models is extremely important to elucidate the capacity of free-living organisms to establish and cause disease in hosts. The aim of this study was to compare and evaluate the histopathology and culture between two different routes of experimental infection of T4 Acanthamoeba isolated from environmental and clinical source in mice (intracranial and intraperitoneal). Swiss isogenic healthy mice were inoculated with 10(4) trophozoites by intracranial (IC) and intraperitoneal (IP) routes and observed during 21 days. The brains from animals inoculated by the IC route were collected and from the animals of the IP inoculation group, the brains, livers, kidneys, spleens, and lungs were removed. The organs were prepared and appropriately divided to be evaluated with histopathology and culture. There was no significant difference between the inoculation routes in terms of isolates recovery (χ(2) = 0.09; p = 0.76). In the IC group, isolate recovery rate was significantly higher in histopathology than the one achieved by culture (χ(2) = 6.45; p < 0.01). Experimental infection revealed that all isolates inoculated could be considered invasive because it was possible to recover evolutive forms of Acanthamoeba in both routes. This work represents the first in vivo pathogenicity assay of primary isolation source in Central region of Brazil showing in vivo pathogenicity and hematogenous spread capacity of these protozoa, improving the knowledge on free-living amoebae isolates.

  16. Evaluation of Acanthamoeba myosin-IC as a potential therapeutic target.

    PubMed

    Martín-Navarro, Carmen M; Lorenzo-Morales, Jacob; López-Arencibia, Atteneri; Reyes-Batlle, María; Piñero, José E; Valladares, Basilio; Maciver, Sutherland K

    2014-01-01

    Members of the genus Acanthamoeba are facultative pathogens of humans, causing a sight-threatening keratitis and a fatal encephalitis. We have targeted myosin-IC by using small interfering RNA (siRNA) silencing as a therapeutic approach, since it is known that the function of this protein is vital for the amoeba. In this work, specific siRNAs against the Acanthamoeba myosin-IC gene were developed. Treated and control amoebae were cultured in growth and encystment media to evaluate the induced effects after myosin-IC gene knockdown, as we have anticipated that cyst formation may be impaired. The effects of myosin-IC gene silencing were inhibition of cyst formation, inhibition of completion of cytokinesis, inhibition of osmoregulation under osmotic stress conditions, and death of the amoebae. The finding that myosin-IC silencing caused incompletion of cytokinesis is in agreement with earlier suggestions that the protein plays a role in cell locomotion, which is necessary to pull daughter cells apart after mitosis in a process known as "traction-mediated cytokinesis". We conclude that myosin-IC is a very promising potential drug target for the development of much-needed antiamoebal drugs and that it should be further exploited for Acanthamoeba therapy.

  17. Presumed late recurrence of Acanthamoeba keratitis exacerbated by exposure to topical corticosteroids

    PubMed Central

    Patel, Dipika V; McGhee, Charles NJ

    2013-01-01

    A 28-year-old female with a history of contact lens wear presented with a 1 week history of pain and photophobia in her left eye. In vivo confocal microscopy (IVCM) and corneal scrape confirmed the diagnosis of Acanthamoeba keratitis (AK) which was treated with intensive topical propamidine isethionate (0.1%) and chlorhexidine (0.02%) with tapering dosage over 11 months. Five years after complete resolution of AK and cessation of all contact lens wear, the subject presented to her optometrist with a history of ocular discomfort and mild photophobia. Without further investigation she was prescribed topical corticosteroids. Three weeks later she presented with pain and reduced vision in the left eye. Slit-lamp examination revealed focal, inferior corneal stromal edema. IVCM confirmed widespread Acanthamoeba cysts. Treatment with topical polyhexamethylene biguanide (PHMB) 0.02% and propamidine isethionate 0.1% resulted in resolution of the AK. Despite an initially mild AK, this subject presumably retained viable Acanthamoeba cysts in her cornea 5 years after the initial episode. This report highlights the importance of caution when using corticosteroids in patients with a previous history of AK, even in the relatively distant past. Patients with AK should be warned regarding the risks of recurrence following presumed resolution. PMID:24391372

  18. Inactivation of Acanthamoeba spp. and Other Ocular Pathogens by Application of Cold Atmospheric Gas Plasma

    PubMed Central

    Shama, Gilbert; Andrew, Peter W.

    2016-01-01

    Currently there are estimated to be approximately 3.7 million contact lens wearers in the United Kingdom and 39.2 million in North America. Contact lens wear is a major risk factor for developing an infection of the cornea known as keratitis due to poor lens hygiene practices. While there is an international standard for testing disinfection methods against bacteria and fungi (ISO 14729), no such guidelines exist for the protozoan Acanthamoeba, which causes a potentially blinding keratitis most commonly seen in contact lens wearers, and as a result, many commercially available disinfecting solutions show incomplete disinfection after 6 and 24 h of exposure. Challenge test assays based on international standard ISO 14729 were used to determine the antimicrobial activity of cold atmospheric gas plasma (CAP) against Pseudomonas aeruginosa, Candida albicans, and trophozoites and cysts of Acanthamoeba polyphaga and Acanthamoeba castellanii. P. aeruginosa and C. albicans were completely inactivated in 0.5 min and 2 min, respectively, and trophozoites of A. polyphaga and A. castellanii were completely inactivated in 1 min and 2 min, respectively. Furthermore, for the highly resistant cyst stage of both species, complete inactivation was achieved after 4 min of exposure to CAP. This study demonstrates that the CAP technology is highly effective against bacterial, fungal, and protozoan pathogens. The further development of this technology has enormous potential, as this approach is able to deliver the complete inactivation of ocular pathogens in minutes, in contrast to commercial multipurpose disinfecting solutions that require a minimum of 6 h. PMID:26994079

  19. Acanthamoeba T4, T5 and T11 isolated from mineral water bottles in southern Brazil.

    PubMed

    Maschio, Vinicius José; Chies, Fernanda; Carlesso, Ana Maris; Carvalho, Amanda; Rosa, Sayonara Peixoto; Van Der Sand, Sueli Teresinha; Rott, Marilise Brittes

    2015-01-01

    Acanthamoeba is a protist potential pathogen, capable of causing a blinding keratitis in contact lens wearers and disseminated infection, leading to granulomatous amebic encephalitis in immunocompromised individuals. This amoeba is a ubiquitous organism that has been isolated from various domestic water systems, such as cooling towers and hospital water networks. The objective of this work was to investigate the presence of Acanthamoeba in mineral water bottles marketed in Porto Alegre, southern Brazil. Positive samples were further classified at the genotype level after sequencing the ASA.S1 region of 18S rDNA gene. Six of the eight isolates belonged to T5 genotype, one to T4 genotype, and one was T11. Several genotypes have been reported worldwide as causative of pathologies in humans, including genotypes T4, T5 and T11. Overall, the widespread distribution of potentially pathogenic Acanthamoeba strains in the studied source demands more awareness within the public and health professionals, because this pathogen is emerging as a risk for human health worldwide.

  20. Photodynamic inactivation of Acanthamoeba polyphaga with curcuminoids: an in vitro study

    NASA Astrophysics Data System (ADS)

    Corrêa, Thaila Q.; Geralde, Mariana C.; Carvalho, Mariana T.; Bagnato, Vanderlei S.; Kurachi, Cristina; de Souza, Clovis W. O.

    2016-03-01

    Acanthamoeba polyphaga are free-living amoebae that can be considered potentially pathogenic organisms by cause serious human infections, including keratitis and granulomatous amoebic encephalitis that usually results in death. Photodynamic inactivation (PDI) has been used for the biological control of microorganisms and can be promise in the control of Acanthamoeba infections. This study evaluated the in vitro effectiveness of PDI in A. polyphaga using curcuminoids salt as photosensitizer (PS) besides observing morphological changes caused by this PS in this organism, in confocal microscopy. A. polyphaga trophozoites were grown at 37°C in PYG medium for 48 to 72 hours. After, the trophozoites were incubated with PS solution during one hour and the samples were irradiated using light-emitting diodes at 460 nm at light doses 30 and 50 J/cm2. The results revealed reduction of 27.7%, 61.4% and 82.5% at 30 J/cm2 and 75.2%, 85.0% and 95.9% at 50 J/cm2, respectively, at curcuminoid salt concentrations of 500, 1000 and 1500 μg/mL. Through fluorescence images, it was possible to visualize the curcuminoid salt's uptake by the trophozoites. The PS showed toxicity to amoebae, in the dark, but the irradiation in PDI contributed to amoebae death effect. These data suggest that PDI may be an application of therapeutic intervention against Acanthamoeba infections, since it was effective in the inactivation of these amoebae.

  1. Acanthamoeba spp. and bacterial contamination in contact lens storage cases and the relationship to user profiles.

    PubMed

    Pens, Claiton José; da Costa, Marisa; Fadanelli, Cristina; Caumo, Karin; Rott, MariliseBrittes

    2008-11-01

    Storage cases for contact lenses receive microbiota from the environment, body, and eye, which can form biofilms. These biofilms, in addition to causing discomfort and cloudy vision, can cause local irritation, facilitate the adherence of microorganisms, and lead to infection. The objective of this study was to evaluate the presence of bacteria and Acanthamoeba spp. in the biofilm and solutions in contact lens storage cases, and to assess their relationships to the habits of contact lens wearers. Eighty-one volunteers assembled from the ophthalmology section of a public hospital and from the Central Campus of the federal university, both in the state of Rio Grande do Sul, Brazil, provided the contact lens storage cases. The samples collected were inoculated into sheep blood agar, to isolate bacteria; and into 1.5% non-nutrient agar with an overlayer of Escherichia coli, to isolate free-living amoebas. Of the 81 samples analyzed, 58 (71%) showed bacterial growth and seven (8.6%) were positive for Acanthamoeba spp. The amoebas were identified according to the morphological criteria of Page (A new key to fresh water and soil gymnamoebae, Freshwater Biology Association, Ambleside, UK, 1988) and confirmed by PCR. The storage cases that were positive for Acanthamoeba spp. had a mean of 10(7) UFC/mL and belonged to individuals who had not taken sufficient care with hand washing.

  2. In vitro effects of selected contact lens care solutions on Acanthamoeba castellanii strains in Poland.

    PubMed

    Padzik, Marcin; Chomicz, Lidia; Szaflik, Jacek P; Chruścikowska, Agnieszka; Perkowski, Konrad; Szaflik, Jerzy

    2014-11-01

    Free-living, cosmopolitan amoebae of the Acanthamoeba genus may be the causative agents of Acanthamoeba keratitis (AK) - a progressive, vision-threatening infection of the human cornea described particularly among contact lens wearers. Use of contact lens care solutions, effective against these organisms, is important in preventing AK infections. 3 different strains of Acanthamoeba castellanii of the T4 genotype (Neff strain and two others, isolated from patients with AK) were exposed to 4 selected multipurpose contact lens care solutions available in Poland: Ciba Vision AoSept Plus, Bausch & Lomb ReNu MultiPlus, Alcon Opti-Free, Ciba Vision Solo Care Aqua. No amoebicidal effect was observed. The strongest amoebostatic effect was visible after 24h of exposition to Opti-Free and ReNu solution and associated with percentage increase of rounded, motionless forms. This is significantly longer than minimum disinfection time recommended by manufacturers of all tested multipurpose solutions. Surprisingly, no clear induction of the encystation process was observed.

  3. Acanthamoeba meningoencephalitis in immunocompetent: A case report and review of literature

    PubMed Central

    Khanna, Vinay; Shastri, BA; Anusha, G; Mukhopadhayay, Chiranjay; Khanna, Ruchee

    2014-01-01

    A 30-year-old manual laborer from Karnataka, India presented with intermittent low grade fever and diffuse headache for 1 month. On examination, patient had enlarged supraclavicular and cervical lymph nodes. Patient had positive Kernig's sign and neck stiffness. Motor, sensory and cranial nerve examinations were within the normal limits. Abdominal, cardiovascular and chest examination did not yield any positive findings. Contrast enhanced computed tomography head was normal. Patient was suspected to have extrapulmonary tuberculosis. Patient was started on antitubercular drugs. Diagnostic lumbar puncture was performed. Wet mount and Giemsa smear preparation of cerebrospinal fluid (CSF) showed trophozoites suggestive of Acanthamoeba. CSF was cultured onto non-nutrient agar with an overlay of Escherichia coli. Wet mount made from the culture media yielded cysts and trophozoites of Acanthamoeba spp. Patient was diagnosed with Acanthamoeba meningitis and was started on specific therapy with Rifampicin 600 mg once a day, Cotrimoxazole 960 mg twice-a-day and Fluconazole 400 mg once daily for 2 weeks. Patient had a complete recovery and was discharged from the hospital. PMID:25250233

  4. In Vitro Efficacy of Corifungin against Acanthamoeba castellanii Trophozoites and Cysts

    PubMed Central

    Debnath, Anjan; Tunac, Josefino B.; Silva-Olivares, Angélica; Galindo-Gómez, Silvia

    2014-01-01

    Painful blinding keratitis and fatal granulomatous amebic encephalitis are caused by the free-living amebae Acanthamoeba spp. Several prescription eye medications are used to treat Acanthamoeba keratitis, but the infection can be difficult to control because of recurrence of infection. For the treatment of encephalitis, no single drug was found useful, and in spite of the use of a combination of multiple drugs, the mortality rate remains high. Therefore, efficient, novel drugs are urgently needed for the treatment of amebic keratitis and granulomatous amebic encephalitis. In this study, we identified corifungin, a water-soluble polyene macrolide, as amebicidal. In vitro, it was effective against both the trophozoites and the cysts. Transmission electron microscopy of Acanthamoeba castellanii incubated with corifungin showed the presence of swollen mitochondria, electron-dense granules, degeneration of cytoplasm architecture, and loss of nuclear chromatin structure. These changes were followed by lysis of amebae. Corifungin also induced the encystment process of A. castellanii. There were alterations in the cyst cell wall followed by lysis of the cysts. Corifungin is a promising therapeutic option for keratitis and granulomatous amebic encephalitis. PMID:24366747

  5. Caspase-like proteins: Acanthamoeba castellanii metacaspase and Dictyostelium discoideum paracaspase, what are their functions?

    PubMed

    Saheb, Entsar; Trzyna, Wendy; Bush, John

    2014-12-01

    Caspases are cysteine proteases that are important regulators of programmed cell death in animals. Two novel relatives to members of the caspase families metacaspases and paracaspase have been discovered. Metacaspase type-1 was identified in Acanthamoeba castellanii, an opportunistic protozoan parasite that causes severe diseases in humans. Paracaspase was found in the non-pathogenic protozoan Dictyostelium discoideum. Since their discovery in Acanthamoeba and Dictyostelium, metacaspases and paracaspases have remained poorly characterized. At present we do not have sufficient data about the molecular function of these caspase-like proteins or their role, if any, in programmed cell death. How these caspase proteins function at the molecular level is an important area of study that will provide insight into their potential for treatment therapies against Acanthamoeba infection and other similar parasitic protozoan. Additionally, finding the molecular functions of these caspase-like proteins will provide information concerning their role in more complex organisms.The aim of this article was to review recent discoveries about metacaspases and paracaspases as regulators of apoptotic and non-apoptotic processes.

  6. Francisella philomiragia biofilm formation and interaction with the aquatic protist Acanthamoeba castellanii.

    PubMed

    Verhoeven, Anne B; Durham-Colleran, Meghan W; Pierson, Tony; Boswell, William T; Van Hoek, Monique L

    2010-10-01

    The bacterium Francisella philomiragia has been isolated from environmental samples originating from around the globe. F. philomiragia-related strains cause francisellosis of both farmed and wild fish. In addition, occasional human infections caused by F. philomiragia are found in victims of near-drowning and patients with chronic granulomatous disease. We have shown that F. philomiragia forms in vitro biofilms with increased formation at 25 °C over 37 °C conditions. We found that F. philomiragia can form a biofilm in a co-culture with live Acanthamoeba castellanii, an aquatic amoeba. Interestingly, amoeba-conditioned supernatant has an inhibitory effect on production of biofilm by F. philomiragia, whereas Francisella-conditioned supernatant has no effect on growth of amoebae. We have shown that F. philomiragia can infect A. castellanii after only 5 days of co-incubation and that it infects A. castellanii more quickly than the related species F. novicida does. Our studies point to a potentially overlooked interaction between F. philomiragia and Acanthamoeba. This relationship in the marine lifecycle of F. philomiragia may support the persistence of the bacterium in waterways and its ability to infect fish. An understanding of the persistence of this organism in aquatic systems through biofilm formation and its interaction with Acanthamoeba will be important in developing prevention strategies for this pathogen.

  7. Prevalence of Acanthamoeba and superbugs in a clinical setting: coincidence or hyperparasitism?

    PubMed

    Siddiqui, Ruqaiyyah; Sagheer, Mehwish; Khan, Naveed Ahmed

    2013-03-01

    Antibacterial strategies to eradicate superbugs from hospitals/nursing homes have had limited success, suggesting the need for employing innovative preventative measures and better understanding of the prevalence of microbial pathogens in close proximity of susceptible populations. A total of 120 environmental samples were collected from the Aga Khan University hospital. Amoebae were identified using morphological characteristics as well as PCR using genus-specific primers, while bacteria were identified using standard biochemical testing. Out of 120 samples tested, 52 (43.3 %) samples were positive for Acanthamoeba, while all 120 (100 %) samples were positive for bacteria. Following bacterial identification, samples showed mixed bacterial populations. Out of 120 samples, 76 (63.3 %) samples were positive for Bacillus spp., 64 (53.3 %) samples were positive for Corynebacterium spp., 32 (26.6 %) samples were positive for Staphylococcus spp., and 9 (7.5 %) samples were positive for Micrococcus spp. The antibiotic susceptibility showed that all bacterial isolates recovered were multiple drug-resistant. The current findings suggest that Acanthamoeba and bacteria coexist in a clinical environment. Given that Acanthamoeba can harbor bacteria, anti-amoebic approaches may represent a strategy in eradicating "superbugs" from the clinical setting in addition to the current measures.

  8. Purification of a protein phosphatase from Acanthamoeba that dephosphorylates and activates myosin II.

    PubMed

    McClure, J A; Korn, E D

    1983-12-10

    The actin-activated ATPase activity of myosin II from Acanthamoeba castellanii is inhibited by phosphorylation of 3 serine residues near the carboxyl end of the heavy chain of the molecule. We have purified a protein phosphatase from Acanthamoeba using myosin II as a substrate. This phosphatase has a molecular weight of 39,000 by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and an isoelectric point in urea of 5.2. The enzyme also is active against other phosphoserine protein substrates such as turkey gizzard smooth muscle myosin light chain, but not against a synthetic phosphotyrosine protein substrate. It does not hydrolyze ATP or p-nitrophenol phosphate. No effector has been found to increase substantially the activity of the enzyme as isolated, but it is inhibited by ATP, pyrophosphate, and NaF. This inhibition is reduced in the presence of MnCl2. The Mg2+-dependent actin-activated ATPase of myosin II is activated by dephosphorylation of phosphorylated myosin II by the phosphatase. Its broad substrate specificity, molecular weight, and response to protein phosphatase inhibitors suggest that the Acanthamoeba protein phosphatase is a type 2A phosphatase (Cohen, P. (1982) Nature (Lond.) 206, 613-620).

  9. Evaluation of Acanthamoeba Myosin-IC as a Potential Therapeutic Target

    PubMed Central

    Lorenzo-Morales, Jacob; López-Arencibia, Atteneri; Reyes-Batlle, María; Piñero, José E.; Valladares, Basilio; Maciver, Sutherland K.

    2014-01-01

    Members of the genus Acanthamoeba are facultative pathogens of humans, causing a sight-threatening keratitis and a fatal encephalitis. We have targeted myosin-IC by using small interfering RNA (siRNA) silencing as a therapeutic approach, since it is known that the function of this protein is vital for the amoeba. In this work, specific siRNAs against the Acanthamoeba myosin-IC gene were developed. Treated and control amoebae were cultured in growth and encystment media to evaluate the induced effects after myosin-IC gene knockdown, as we have anticipated that cyst formation may be impaired. The effects of myosin-IC gene silencing were inhibition of cyst formation, inhibition of completion of cytokinesis, inhibition of osmoregulation under osmotic stress conditions, and death of the amoebae. The finding that myosin-IC silencing caused incompletion of cytokinesis is in agreement with earlier suggestions that the protein plays a role in cell locomotion, which is necessary to pull daughter cells apart after mitosis in a process known as “traction-mediated cytokinesis”. We conclude that myosin-IC is a very promising potential drug target for the development of much-needed antiamoebal drugs and that it should be further exploited for Acanthamoeba therapy. PMID:24468784

  10. Anaerobic respiration: In vitro efficacy of Nitazoxanide against mitochondriate Acanthamoeba castellanii of the T4 genotype.

    PubMed

    Aqeel, Yousuf; Siddiqui, Ruqaiyyah; Farooq, Maria; Khan, Naveed Ahmed

    2015-10-01

    Acanthamoeba is an opportunistic protist pathogen that is responsible for serious human and animal infection. Being one of the most frequently isolated protists from the environment, it is likely that it readily encounters microaerophilic environments. For respiration under anaerobic or low oxygen conditions in several amitochondriate protists, decarboxylation of pyruvate is catalyzed by pyruvate ferredoxin oxidoreductase instead of pyruvate dehydrogenase. In support, Nitazoxanide, an inhibitor of pyruvate ferredoxin oxidoreductase, is effective and non-mutagenic clinically against a range of amitochondriate protists, Giardia intestinalis, Entamoeba histolytica and Trichomonas vaginalis. The overall aim of the present study was to determine in vitro efficacy of Nitazoxanide against Acanthamoeba castellanii. At micromolar concentrations, the findings revealed that Nitazoxanide neither affected A. castellanii growth or viability nor amoeba-mediated host cell monolayer damage in vitro or extracellular proteolytic activities. Similarly, microaerophilic conditions alone had no significant effects. In contrast, microaerophilic conditions together with Nitazoxanide showed amoebicidal effects and inhibited A. castellanii-mediated host cell monolayer damage as well as extracellular proteases. Using encystation assays, it was observed that Nitazoxanide inhibited trophozoite transformation into cysts both under aerophilic and microaerophilic conditions. Furthermore, pre-treatment of cysts with Nitazoxanide inhibited A. castellanii excystation. These findings are important in the identification of potential targets that could be useful against parasite-specific respiration as well as to understand the basic biology of the life cycle of Acanthamoeba.

  11. Genotypic, phenotypic, biochemical, physiological and pathogenicity-based categorisation of Acanthamoeba strains.

    PubMed

    Khan, Naveed Ahmed; Tareen, Noor Khan

    2003-06-01

    The genus Acanthamoeba includes more than 20 morphological species, but classification is problematical. Recently, the discovery of substantial interstrain differences in ribosomal DNA (rDNA) sequences has prompted questions about the relatedness of strains of the same species. In this study, therefore, we have investigated relationships between two isolates of A. polyphaga, CCAP 1501/3c and ATCC 30871, using morphological, biochemical, physiological, molecular and cytotoxicity assays. We observed that A. polyphaga ATCC 30871 exhibited up to six arms in endocyst while A. polyphaga CCAP 1501/3c exhibited a maximum of 5 arms thus indicating their position in group 2 and 3, respectively. Acanthamoeba polyphaga ATCC 30871 exhibited growth at 37 degrees C and growth on 1M mannitol plates while A. polyphaga CCAP 1501/3c did not. In addition, both isolates exhibited differences in isoenzyme banding patterns and rDNA restriction fragment polymorphisms. More importantly, A. polyphaga ATCC 30871 produced cytotoxicity on corneal epithelial cells while A. polyphaga CCAP 1501/3c had no effects, suggesting differences in pathogenicity. Thus, all the results provide evidence for significant differences between the strains and further provided the basis for reclassification of the isolates. Implications of these results in the clinical diagnosis of pathogenic Acanthamoeba are discussed.

  12. Comparative proteomic analysis of extracellular secreted proteins expressed by two pathogenic Acanthamoeba castellanii clinical isolates and a non-pathogenic ATCC strain.

    PubMed

    Huang, Jian-Ming; Lin, Wei-Chen; Li, Sung-Chou; Shih, Min-Hsiu; Chan, Wen-Ching; Shin, Jyh-Wei; Huang, Fu-Chin

    2016-07-01

    Acanthamoeba keratitis (AK) is a serious ocular disease caused by pathogenic Acanthamoeba gaining entry through wounds in the corneal injury; generally, patients at risk for contracting AK wear contact lenses, usually over a long period of time. Moreover, pathogenic Acanthamoeba causes serious consequences: it makes the cornea turbid and difficult to operate on, including procedures such as enucleation of the eyeball. At present, diagnosis of this disease is not straightforward, and treatment is very demanding. We have established the comparative transcriptome and extracellular secreted proteomic database according to the non-pathogenic strain ATCC 30010 and the pathogenic strains NCKU_B and NCKU_D. We identified 44 secreted proteins successfully, 10 consensus secreted proteins and 34 strain-specific secreted proteins. These proteins may provide targets for therapy and immuno-diagnosis of Acanthamoeba infections. This study shows a suitable approach to identify secreted proteins in Acanthamoeba and provides new perspectives for the study of molecules potentially involved in the AK.

  13. High occurrence of Acanthamoeba genotype T4 in soil sources from Bolívar State, Venezuela.

    PubMed

    Wagner, Carolina; Reyes-Batlle, María; Hernán, Aurora; Rojas, Elsy; Pérez, Gladymar; López-Arencibia, Atteneri; Sifaoui, Ines; Martínez-Carretero, Enrique; Piñero, José E; Valladares, Basilio; Lorenzo-Morales, Jacob

    2016-09-01

    Pathogenic strains of Acanthamoeba are causative agents of keratitis and encephalitis that often may end fatal in humans and other animals. In the present study, twenty-seven soil samples were collected in the Bolivar State in Venezuela and checked for the presence of Acanthamoeba. Samples were cultivated onto 2% non-nutrient agar plates seeded with a layer of heat killed E. coli. Amplification by PCR and sequencing of the DF3 region of the 18S rDNA of Acanthamoeba was carried out in order to confirm morphological identification of the amoebae. Furthermore, Acanthamoeba spp. was isolated from 51.8% of soil samples. Sequencing of the DF3 region of the 18S rDNA resulted in the identification of genotype T4 in all samples. To the best of our knowledge, this is the first report of genotype T4 in soil sources from Venezuela. Further studies should be carried out in this State and in the country in order to determine the current occurrence of Acanthamoeba in Venezuelan environments.

  14. Characterization of a new pathogenic Acanthamoeba Species, A. byersi n. sp., isolated from a human with fatal amoebic encephalitis.

    PubMed

    Qvarnstrom, Yvonne; Nerad, Thomas A; Visvesvara, Govinda S

    2013-01-01

    Acanthamoeba spp. are free-living amoebae that are ubiquitous in natural environments. They can cause cutaneous, nasopharyngeal, and disseminated infection, leading to granulomatous amebic encephalitis (GAE) in immunocompromised individuals. In addition, they can cause amoebic keratitis in contact lens wearers. Acanthamoeba GAE is almost always fatal because of difficulty and delay in diagnosis and lack of optimal antimicrobial therapy. Here, we report the description of an unusual strain isolated from skin and brain of a GAE patient. The amoebae displayed large trophozoites and star-shaped cysts, characteristics for acanthamoebas belonging to morphology Group 1. However, its unique morphology and growth characteristics differentiated this new strain from other Group 1 species. DNA sequence analysis, secondary structure prediction, and phylogenetic analysis of the 18S rRNA gene confirmed that this new strain belonged to Group 1, but that it was distinct from the other sequence types within that group. Thus, we hereby propose the establishment of a new species, Acanthamoeba byersi n. sp. as well as a new sequence type, T18, for this new strain. To our knowledge, this is the first report of a Group 1 Acanthamoeba that is indisputably pathogenic in humans.

  15. Synthesis, characterization and amoebicidal potential of locally synthesized TiO2 nanoparticles against pathogenic Acanthamoeba trophozoites in vitro.

    PubMed

    Imran, Muhammad; Muazzam, Ambreen Gul; Habib, Amir; Matin, Abdul

    2016-06-01

    Acanthamoeba is an opportunistic protozoan pathogen that plays a pivotal role in the ecosystem. It may cause blinding keratitis and fatal encephalitis involving the central nervous system. Here we synthesized pure and Zn doped TiO2 nanoparticles (~10-30nm) via sol-gel and sol-hydrothermal methods and demonstrated its impact on the biological characteristics of pathogenic Acanthamoeba castellanii. Our results revealed that pure and Zn doped TiO2 nanoparticles synthesized by sol-hydrothermal methods (ranging 5, 10, 25 and 50μg/ml) exhibited amoebicidal effects i.e., >60% of trophozoites executed under normal light at maximum dose (50μg/ml) within 1h incubation. In contrast pure/doped TiO2 obtained via sol gel method showed ~40% amoeba damage. Furthermore, amoebae growth assay demonstrated that Zn doped TiO2 also inhibited Acanthamoeba numbers up to 7days in dose dependent manner. It was interesting to note that all the tested TiO2 nanoparticles have shown maximum amoebicidal effects at pH7 which is quite relevant to amoebic growth favorable conditions. Our results confirmed that TiO2 has inhibitory effects on Acanthamoeba growth and viability. Overall, we reported the amoebicidal and amoebic growth inhibition potential of pure and Zn doped TiO2 nanoparticles against Acanthamoeba due to attached OH(-) groups, reduced size and decreased band gap of sol hydrothermally synthesized TiO2 nanoparticles.

  16. Induction of morphological and electrophysiological changes in hamster cornea after in vitro interaction with trophozoites of Acanthamoeba spp.

    PubMed

    Omaña-Molina, Maritza; Navarro-García, Fernando; González-Robles, Arturo; Serrano-Luna, José de Jesús; Campos-Rodríguez, Rafael; Martínez-Palomo, Adolfo; Tsutsumi, Víctor; Shibayama, Mineko

    2004-06-01

    Acanthamoeba castellani and Acanthamoeba polyphaga are free-living amebae that cause keratitis and granulomatous encephalitis in humans. We have analyzed the early morphological and electrophysiological changes occurring during the in vitro interaction of cultured amebae with intact or physically damaged corneas obtained from hamsters. Both species of Acanthamoeba produced similar cytopathic changes, as seen by light microscopy and scanning electron microscopy. After adhesion to the epithelial surface, trophozoites formed clumps and migrated toward the cell borders, causing the separation of adjacent cells at 1 h of coculture. At later stages (2 to 4 h), some amebae were found under desquamating epithelial cells whereas others were seen associated with damaged cells or forming amebostome-like structures to ingest detached epithelial cells. Control corneas incubated in culture medium conditioned with amebae showed a cytoplasmic vacuolization and blurring of the epithelial-stromal junction. The early stages of corneal epithelial damage caused by amebae were also analyzed by measuring the transepithelial resistance changes in corneas mounted in Ussing chambers. Both species of Acanthamoeba caused a rapid decrease in electrical resistance. The present observations demonstrate that under in vitro conditions, Acanthamoeba trophozoites rapidly cause significant damage to the corneal epithelium. Furthermore, in our experimental model, previous physical damage to the corneas was not a prerequisite for the development of amebic corneal ulcerations.

  17. Induction of Morphological and Electrophysiological Changes in Hamster Cornea after In Vitro Interaction with Trophozoites of Acanthamoeba spp.

    PubMed Central

    Omaña-Molina, Maritza; Navarro-García, Fernando; González-Robles, Arturo; Serrano-Luna, José de Jesús; Campos-Rodríguez, Rafael; Martínez-Palomo, Adolfo; Tsutsumi, Víctor; Shibayama, Mineko

    2004-01-01

    Acanthamoeba castellani and Acanthamoeba polyphaga are free-living amebae that cause keratitis and granulomatous encephalitis in humans. We have analyzed the early morphological and electrophysiological changes occurring during the in vitro interaction of cultured amebae with intact or physically damaged corneas obtained from hamsters. Both species of Acanthamoeba produced similar cytopathic changes, as seen by light microscopy and scanning electron microscopy. After adhesion to the epithelial surface, trophozoites formed clumps and migrated toward the cell borders, causing the separation of adjacent cells at 1 h of coculture. At later stages (2 to 4 h), some amebae were found under desquamating epithelial cells whereas others were seen associated with damaged cells or forming amebostome-like structures to ingest detached epithelial cells. Control corneas incubated in culture medium conditioned with amebae showed a cytoplasmic vacuolization and blurring of the epithelial-stromal junction. The early stages of corneal epithelial damage caused by amebae were also analyzed by measuring the transepithelial resistance changes in corneas mounted in Ussing chambers. Both species of Acanthamoeba caused a rapid decrease in electrical resistance. The present observations demonstrate that under in vitro conditions, Acanthamoeba trophozoites rapidly cause significant damage to the corneal epithelium. Furthermore, in our experimental model, previous physical damage to the corneas was not a prerequisite for the development of amebic corneal ulcerations. PMID:15155626

  18. Inhibition of 3-hydroxy-3-methylglutaryl-coenzyme A reductase and application of statins as a novel effective therapeutic approach against Acanthamoeba infections.

    PubMed

    Martín-Navarro, Carmen María; Lorenzo-Morales, Jacob; Machin, Rubén P; López-Arencibia, Atteneri; García-Castellano, José Manuel; de Fuentes, Isabel; Loftus, Brendan; Maciver, Sutherland K; Valladares, Basilio; Piñero, José E

    2013-01-01

    Acanthamoeba is an opportunistic pathogen in humans, whose infections most commonly manifest as Acanthamoeba keratitis or, more rarely, granulomatous amoebic encephalitis. Although there are many therapeutic options for the treatment of Acanthamoeba, they are generally lengthy and/or have limited efficacy. Therefore, there is a requirement for the identification, validation, and development of novel therapeutic targets against these pathogens. Recently, RNA interference (RNAi) has been widely used for these validation purposes and has proven to be a powerful tool for Acanthamoeba therapeutics. Ergosterol is one of the major sterols in the membrane of Acanthamoeba. 3-Hydroxy-3-methylglutaryl-coenzyme A (HMG-CoA) reductase is an enzyme that catalyzes the conversion of HMG-CoA to mevalonate, one of the precursors for the production of cholesterol in humans and ergosterol in plants, fungi, and protozoa. Statins are compounds which inhibit this enzyme and so are promising as chemotherapeutics. In order to validate whether this enzyme could be an interesting therapeutic target in Acanthamoeba, small interfering RNAs (siRNAs) against HMG-CoA were developed and used to evaluate the effects induced by the inhibition of Acanthamoeba HMG-CoA. It was found that HMG-CoA is a potential drug target in these pathogenic free-living amoebae, and various statins were evaluated in vitro against three clinical strains of Acanthamoeba by using a colorimetric assay, showing important activities against the tested strains. We conclude that the targeting of HMG-CoA and Acanthamoeba treatment using statins is a novel powerful treatment option against Acanthamoeba species in human disease.

  19. Inhibition of 3-Hydroxy-3-Methylglutaryl–Coenzyme A Reductase and Application of Statins as a Novel Effective Therapeutic Approach against Acanthamoeba Infections

    PubMed Central

    Lorenzo-Morales, Jacob; Machin, Rubén P.; López-Arencibia, Atteneri; García-Castellano, José Manuel; de Fuentes, Isabel; Loftus, Brendan; Maciver, Sutherland K.; Valladares, Basilio; Piñero, José E.

    2013-01-01

    Acanthamoeba is an opportunistic pathogen in humans, whose infections most commonly manifest as Acanthamoeba keratitis or, more rarely, granulomatous amoebic encephalitis. Although there are many therapeutic options for the treatment of Acanthamoeba, they are generally lengthy and/or have limited efficacy. Therefore, there is a requirement for the identification, validation, and development of novel therapeutic targets against these pathogens. Recently, RNA interference (RNAi) has been widely used for these validation purposes and has proven to be a powerful tool for Acanthamoeba therapeutics. Ergosterol is one of the major sterols in the membrane of Acanthamoeba. 3-Hydroxy-3-methylglutaryl–coenzyme A (HMG-CoA) reductase is an enzyme that catalyzes the conversion of HMG-CoA to mevalonate, one of the precursors for the production of cholesterol in humans and ergosterol in plants, fungi, and protozoa. Statins are compounds which inhibit this enzyme and so are promising as chemotherapeutics. In order to validate whether this enzyme could be an interesting therapeutic target in Acanthamoeba, small interfering RNAs (siRNAs) against HMG-CoA were developed and used to evaluate the effects induced by the inhibition of Acanthamoeba HMG-CoA. It was found that HMG-CoA is a potential drug target in these pathogenic free-living amoebae, and various statins were evaluated in vitro against three clinical strains of Acanthamoeba by using a colorimetric assay, showing important activities against the tested strains. We conclude that the targeting of HMG-CoA and Acanthamoeba treatment using statins is a novel powerful treatment option against Acanthamoeba species in human disease. PMID:23114753

  20. 18S Ribosomal DNA Typing and Tracking of Acanthamoeba Species Isolates from Corneal Scrape Specimens, Contact Lenses, Lens Cases, and Home Water Supplies of Acanthamoeba Keratitis Patients in Hong Kong

    PubMed Central

    Booton, G. C.; Kelly, D. J.; Chu, Y.-W.; Seal, D. V.; Houang, E.; Lam, D. S. C.; Byers, T. J.; Fuerst, P. A.

    2002-01-01

    We examined partial 18S ribosomal DNA (Rns) sequences of Acanthamoeba isolates cultured in a study of microbial keratitis in Hong Kong. Sequence differences were sufficient to distinguish closely related strains and were used to examine links between strains obtained from corneal scrape specimens, contact lenses, lens cases, lens case solutions, and home water-supply faucets of patients with Acanthamoeba. We also looked for evidence of mixed infections. Identification of Acanthamoeba Rns genotypes was based on sequences of ∼113 bp within the genus-specific amplicon ASA.S1. This permitted genotype identification by using nonaxenic cultures. Of 13 specimens obtained from corneal scrapes, contact lenses, lens cases, or lens case solutions, 12 were Rns genotype T4 and the remaining one was Rns genotype T3. The sequences of corneal scrape specimens of two patients also were the same as those obtained from their contact lenses or lens case specimens. A possible triple-strain infection was indicated by three different T4 sequences in cultures from one patient's lenses. Although faucet water used by patients to clean their lenses is a possible source of infections, specimens isolated from the faucets at two Acanthamoeba keratitis patients' homes differed from their corneal scrape or lens specimens. The overall results demonstrate the potential of this Rns region for tracking Acanthamoeba keratitis strains in infections and for distinguishing single-strain and closely related multiple-strain infections even when other microorganisms might be present with the cultured specimens. They also confirm the predominance of Rns genotype T4 strains in Acanthamoeba keratitis infections. PMID:11980931

  1. 18S ribosomal DNA typing and tracking of Acanthamoeba species isolates from corneal scrape specimens, contact lenses, lens cases, and home water supplies of Acanthamoeba keratitis patients in Hong Kong.

    PubMed

    Booton, G C; Kelly, D J; Chu, Y-W; Seal, D V; Houang, E; Lam, D S C; Byers, T J; Fuerst, P A

    2002-05-01

    We examined partial 18S ribosomal DNA (Rns) sequences of Acanthamoeba isolates cultured in a study of microbial keratitis in Hong Kong. Sequence differences were sufficient to distinguish closely related strains and were used to examine links between strains obtained from corneal scrape specimens, contact lenses, lens cases, lens case solutions, and home water-supply faucets of patients with Acanthamoeba. We also looked for evidence of mixed infections. Identification of Acanthamoeba Rns genotypes was based on sequences of approximately 113 bp within the genus-specific amplicon ASA.S1. This permitted genotype identification by using nonaxenic cultures. Of 13 specimens obtained from corneal scrapes, contact lenses, lens cases, or lens case solutions, 12 were Rns genotype T4 and the remaining one was Rns genotype T3. The sequences of corneal scrape specimens of two patients also were the same as those obtained from their contact lenses or lens case specimens. A possible triple-strain infection was indicated by three different T4 sequences in cultures from one patient's lenses. Although faucet water used by patients to clean their lenses is a possible source of infections, specimens isolated from the faucets at two Acanthamoeba keratitis patients' homes differed from their corneal scrape or lens specimens. The overall results demonstrate the potential of this Rns region for tracking Acanthamoeba keratitis strains in infections and for distinguishing single-strain and closely related multiple-strain infections even when other microorganisms might be present with the cultured specimens. They also confirm the predominance of Rns genotype T4 strains in Acanthamoeba keratitis infections.

  2. Acanthamoeba and mimivirus interactions: the role of amoebal encystment and the expansion of the 'Cheshire Cat' theory.

    PubMed

    Silva, Ludmila Karen Dos Santos; Boratto, Paulo Victor Miranda; La Scola, Bernard; Bonjardim, Cláudio Antônio; Abrahão, Jônatas Santos

    2016-06-01

    Acanthamoeba are natural hosts for giant viruses and their life cycle comprises two stages: a trophozoite and a cryptobiotic cyst. Encystment involves a massive turnover of cellular components under molecular regulation. Giant viruses are able to infect only the trophozoite, while cysts are resistant to infection. Otherwise, upon infection, mimiviruses are able to prevent encystment. This review highlights the important points of Acanthamoeba and giant virus interactions regarding the encystment process. The existence of an acanthamoebal non-permissive cell for Acanthamoeba polyphaga mimivirus, the prototype member of the Mimivirus genus, is analyzed at the molecular and ecological levels, and compared to a similar phenomenon previously described for Emiliana huxleyi and its associated phycodnaviruses: the 'Cheshire Cat' escape strategy.

  3. The use of dimethyl sulfoxide in contact lens disinfectants is a potential preventative strategy against contracting Acanthamoeba keratitis.

    PubMed

    Siddiqui, Ruqaiyyah; Aqeel, Yousuf; Khan, Naveed Ahmed

    2016-10-01

    Acanthamoeba castellanii is the causative agent of blinding keratitis. Though reported in non-contact lens wearers, it is most frequently associated with improper use of contact lens. For contact lens wearers, amoebae attachment to the lens is a critical first step, followed by amoebae binding to the corneal epithelial cells during extended lens wear. Acanthamoeba attachment to surfaces (biological or inert) and migration is an active process and occurs during the trophozoite stage. Thus retaining amoebae in the cyst stage (dormant form) offers an added preventative measure in impeding parasite traversal from the contact lens onto the cornea. Here, we showed that as low as 3% DMSO, abolished A. castellanii excystation. Based on the findings, it is proposed that DMSO should be included in the contact lens disinfectants as an added preventative strategy against contracting Acanthamoeba keratitis.

  4. Prevalence of Acanthamoeba spp. and other free-living amoebae in household water, Ohio, USA--1990-1992.

    PubMed

    Stockman, Lauren J; Wright, Carolyn J; Visvesvara, Govinda S; Fields, Barry S; Beach, Michael J

    2011-03-01

    Knowledge of the prevalence of free-living amoebae (FLA) in US household water can provide a focus for prevention of amoeba-associated illnesses. Household water samples from two Ohio counties, collected and examined for amoebae during 1990-1992, were used to describe the prevalence of Acanthamoeba and other FLA in a household setting. Amoebae were isolated and identified by morphologic features. A total of 2,454 samples from 467 households were examined. Amoebae were found in water samples of 371 (79%) households. Sites most likely to contain amoeba were shower heads (52%) and kitchen sprayers (50%). Species of Hartmannella, Acanthamoeba, or Vahlkampfia were most common. Detection was higher in biofilm swab samples than in water samples. Detection of FLA and Acanthamoeba, at 79% and 51%, respectively, exceed estimates that have been published in previous surveys of household sources. We believe FLA are commonplace inhabitants of household water in this sample as they are in the environment.

  5. The role of domestic tap water on Acanthamoeba keratitis in non-contact lens wearers and validation of laboratory methods.

    PubMed

    Koltas, Ismail Soner; Eroglu, Fadime; Erdem, Elif; Yagmur, Meltem; Tanır, Ferdi

    2015-09-01

    Acanthamoeba is increasingly recognized as an important cause of keratitis in non-contact lens wearers while contact lens wear is the leading risk factor for Acanthamoeba keratitis (AK). It is unlikely that the Acanthamoeba colonization is a feature which is effective only in patient's homes with infectious keratitis since the organism has been isolated from domestic tap water. Two hundred and thirty-one (231) corneal scrapings were taken from infectious keratitis cases, and four contact lens solutions and domestic tap waters were taken from 22 out of 44 AK-diagnosed patient's homes. Microscopic examination, culture, PCR, real-time PCR and DNA sequencing analyses were used for AK-diagnosed samples. The real-time PCR was the most sensitive (100 %) one among the methods used in diagnosis of AK. The 44 (19.0 %) out of 231 corneal scrapings, 4/4 (100 %) contact lens solution and 11/22 (50 %) of domestic tap water samples were found to be positive by real-time PCR for Acanthamoeba. A. griffini (T3), A. castellanii (T4) and A. jacobsi (T15) genotypes were obtained from corneal scrapings, contact lens solutions and domestic tap water samples taken from the patient's homes diagnosed with AK. The isolation of Acanthamoeba containing 6/22 (27.3 %) A. griffini (T3), 14/22 (63.6 %) A. castellanii (T4) and 2/22 (9.1 %) A. jacobsi (T15) from the domestic tap water outlets of 22 of 44 (50 %) of patient's homes revealed that is a significant source of these organisms. A. griffini (T3) and A. jacobsi (T15) genotypes have not been determined from AK cases in Turkey previously. Thus, we conclude that Acanthamoeba keratitis is associated with exposition of patients who has ocular trauma or ocular surface disease to domestic tap water in endemic or potentially endemic countries.

  6. Pathogenic Acanthamoeba spp. Secrete a Mannose-Induced Cytolytic Protein That Correlates with the Ability To Cause Disease

    PubMed Central

    Hurt, Michael; Neelam, Sudha; Niederkorn, Jerry; Alizadeh, Hassan

    2003-01-01

    The pathogenesis of Acanthamoeba keratitis begins when Acanthamoeba trophozoites bind specifically to mannosylated glycoproteins upregulated on the surfaces of traumatized corneal epithelial cells. When Acanthamoeba castellanii trophozoites are grown in methyl-α-d-mannopyranoside, they are induced to secrete a novel 133-kDa protein that is cytolytic to corneal epithelial cells. Clinical isolates of Acanthamoeba spp., and not the soil isolates, were proficient at producing a mannose-induced protein (MIP-133) and generating disease in Chinese hamsters. The purified protein was efficient at killing corneal epithelial cells, the first mechanistic barrier, by inducing apoptosis in a caspase 3-dependent pathway. Subsequent steps in pathogenesis require the amoebae to penetrate and degrade collagen. Only the clinical isolates tested were efficient at migrating through a collagenous matrix in vitro, presumably by MIP-133 degradation of both human type I and human type IV collagen. A chicken anti-MIP-133 antiserum effectively bound to the protein and blocked collagenolytic activity, migration, and cytopathic effects (CPE) against corneal cells in vitro. Chinese hamsters orally immunized with MIP-133 displayed a >30% reduction in disease. Immunoglobulin A isolated from immunized animals bound MIP-133 and blocked CPE on corneal cells in vitro. Animals induced to generate severe chronic infections displayed significant reductions in disease symptoms upon oral immunization postinfection. These data suggest that MIP-133 production might be necessary to initiate corneal disease and that it may play an important role in the subsequent steps of the pathogenic cascade of Acanthamoeba keratitis. Furthermore, as antibodies produced both prior to and after infection reduced clinical symptoms of disease, the protein may represent an important immunotherapeutic target for Acanthamoeba keratitis. PMID:14573643

  7. Pathogenic Acanthamoeba spp secrete a mannose-induced cytolytic protein that correlates with the ability to cause disease.

    PubMed

    Hurt, Michael; Neelam, Sudha; Niederkorn, Jerry; Alizadeh, Hassan

    2003-11-01

    The pathogenesis of Acanthamoeba keratitis begins when Acanthamoeba trophozoites bind specifically to mannosylated glycoproteins upregulated on the surfaces of traumatized corneal epithelial cells. When Acanthamoeba castellanii trophozoites are grown in methyl-alpha-D-mannopyranoside, they are induced to secrete a novel 133-kDa protein that is cytolytic to corneal epithelial cells. Clinical isolates of Acanthamoeba spp., and not the soil isolates, were proficient at producing a mannose-induced protein (MIP-133) and generating disease in Chinese hamsters. The purified protein was efficient at killing corneal epithelial cells, the first mechanistic barrier, by inducing apoptosis in a caspase 3-dependent pathway. Subsequent steps in pathogenesis require the amoebae to penetrate and degrade collagen. Only the clinical isolates tested were efficient at migrating through a collagenous matrix in vitro, presumably by MIP-133 degradation of both human type I and human type IV collagen. A chicken anti-MIP-133 antiserum effectively bound to the protein and blocked collagenolytic activity, migration, and cytopathic effects (CPE) against corneal cells in vitro. Chinese hamsters orally immunized with MIP-133 displayed a >30% reduction in disease. Immunoglobulin A isolated from immunized animals bound MIP-133 and blocked CPE on corneal cells in vitro. Animals induced to generate severe chronic infections displayed significant reductions in disease symptoms upon oral immunization postinfection. These data suggest that MIP-133 production might be necessary to initiate corneal disease and that it may play an important role in the subsequent steps of the pathogenic cascade of Acanthamoeba keratitis. Furthermore, as antibodies produced both prior to and after infection reduced clinical symptoms of disease, the protein may represent an important immunotherapeutic target for Acanthamoeba keratitis.

  8. Analysis of focusing error signals by differential astigmatic method under off-center tracking in the land-groove-type optical disk

    NASA Astrophysics Data System (ADS)

    Shinoda, Masahisa; Nakatani, Hidehiko

    2015-04-01

    We theoretically calculate the behavior of the focusing error signal in the land-groove-type optical disk when the objective lens traverses on out of the radius of the optical disk. The differential astigmatic method is employed instead of the conventional astigmatic method for generating the focusing error signals. The signal behaviors are compared and analyzed in terms of the gain difference of the slope sensitivity of the focusing error signals from the land and the groove. In our calculation, the format of digital versatile disc-random access memory (DVD-RAM) is adopted as the land-groove-type optical disk model, and advantageous conditions for suppressing the gain difference are investigated. The calculation method and results described in this paper will be reflected in the next generation land-groove-type optical disks.

  9. An auto-tuning method for focusing and astigmatism correction in HAADF-STEM, based on the image contrast transfer function.

    PubMed

    Baba, N; Terayama, K; Yoshimizu, T; Ichise, N; Tanaka, N

    2001-01-01

    An auto-tuning method for high-angle annular detector dark field scanning transmission electron microscopy (HAADF-STEM) is proposed which corrects the defocus to the optimum Scherzer focus and compensates the astigmatism. Because the method is based on the image contrast transfer function formulated for the HAADF-STEM, the defocus and the astigmatism are accurately measured from input of two different defocus images. The method is designed to work independent of object function in the linear imaging model by analysing the spectral ratio between two Fourier spectra of their images, which is useful for cases where the spectrum of object function is not uniformly spread out over the reciprocal space. The method was preliminarily tested in a Hitachi HD-2000 STEM, and successful results of the auto-tunings from the viewpoint of verification of the algorithm were obtained using general specimens of Au fine particles and a thin section of a semiconductor device.

  10. Efficacy of Wavefront-guided Photorefractive Keratectomy with Iris Registration for Management of Moderate to High Astigmatism by Advanced Personalized Treatment Nomogram

    PubMed Central

    Mohammadpour, Mehrdad; Hashemi, Hassan; Jabbarvand, Mahmoud; Rahmatnejad, Kamran; Sabet, Fatemeh Alsadat

    2016-01-01

    Purpose: To evaluate the efficacy of photorefractive keratectomy (PRK) using the advanced personalized treatment (APT) nomogram for correction of moderate to high astigmatism. Methods: This prospective interventional case series included 60 consecutive eyes of 30 patients undergoing wavefront-guided PRK (Zyoptix 217 Z100 excimer laser, Bausch & Lomb, Munich, Germany) using the APT nomogram and iris registration for myopic astigmatism. Mitomycin-C was applied intraoperatively in all eyes. Ophthalmic examination was performed preoperatively and 1, 3 and 6 months postoperatively. Results: Preoperatively, mean sphere was -1.68 ± 2.08 diopters (D), mean refractive astigmatism was -3.04 ± 1.05 D and mean spherical equivalent (SE) was -3.12 ± 1.77 D. Six months postoperatively, mean sphere was + 0.60 ± 0.64 D (P < 0.005), mean cylinder was -0.43 ± 0.46 D (P < 0.005) and mean SE was + 0.28 ± 0.48 D (P < 0.005). Hyperopic overcorrection (≥ +1.0 D) occurred in 3 (5%) eyes. Postoperatively, root mean square (RMS) of higher order aberrations (HOAs) was significantly increased (P = 0.041). RMS of spherical aberration (Z [4, 0]) showed no significant change after surgery (P = 0.972). Conclusion: Considering the acceptable residual refractive error, low rate of hyperopic overcorrection, acceptable uncorrected visual acuity, and low risk of postoperative corneal haze, PRK using the APT nomogram with iris registration and mitomycin-C use is a safe and effective modality for treatment of moderate to high astigmatism. PMID:27413491

  11. Anti-drift and auto-alignment mechanism for an astigmatic atomic force microscope system based on a digital versatile disk optical head.

    PubMed

    Hwu, E-T; Illers, H; Wang, W-M; Hwang, I-S; Jusko, L; Danzebrink, H-U

    2012-01-01

    In this work, an anti-drift and auto-alignment mechanism is applied to an astigmatic detection system (ADS)-based atomic force microscope (AFM) for drift compensation and cantilever alignment. The optical path of the ADS adopts a commercial digital versatile disc (DVD) optical head using the astigmatic focus error signal. The ADS-based astigmatic AFM is lightweight, compact size, low priced, and easy to use. Furthermore, the optical head is capable of measuring sub-atomic displacements of high-frequency AFM probes with a sub-micron laser spot (~570 nm, FWHM) and a high-working bandwidth (80 MHz). Nevertheless, conventional DVD optical heads suffer from signal drift problems. In a previous setup, signal drifts of even thousands of nanometers had been measured. With the anti-drift and auto-alignment mechanism, the signal drift is compensated by actuating a voice coil motor of the DVD optical head. A nearly zero signal drift was achieved. Additional benefits of this mechanism are automatic cantilever alignment and simplified design.

  12. Acanthamoeba spp. as a universal host for pathogenic microorganisms: One bridge from environment to host virulence.

    PubMed

    Guimaraes, Allan J; Gomes, Kamilla Xavier; Cortines, Juliana Reis; Peralta, José Mauro; Peralta, Regina H Saramago

    2016-12-01

    Free-living amoebas (FLA) are ubiquitous environmental protists that have enormously contributed to the microbiological contamination of water sources. FLAs have displayed resistance to environmental adversities and germicides and have played important roles in the population control of microbial communities due to its predatory behavior and microbicidal activity. However, some organisms have developed resistance to the intracellular milieu of amoebas, as in the case of Acanthamoebas, which in turn, have been functioning as excellent reservoirs for amoeba-resistant microorganisms (ARMs), such as bacteria, viruses and fungi. Little is known about these relationships and interaction mechanisms, but it is speculated that the FLAs need a very broad repertoire or universal class of receptors to bind and recognize these diverse species of microorganisms. By harboring these organisms as a "Trojan Horse", the Achantamoeba has been working as an excellent vector for pathogens. Moreover, studies have demonstrated that the interaction of pathogens with Acanthamoeba results in environmental selective pressure responsible for induction and maintenance of virulence factors and increase in microbial pathogenicity. This phenomenon is correlated to the observation of higher gene number and DNA content of ARMs, when compared to their relatives which are adapted to other hosts, due to allopatric or sympatric gene transfer and acquisition, contradicting the overall genome reduction theory for intracellularly adapted pathogens. Thus, adaptation to FLAs indirectly provided a "learning" environment for pathogens to resist later to macrophages; besides the evolutionary distance, these phagocytes share similar predatory mechanisms, such as phagocytosis and phagolysossomal degradation. In this mini-review, we cover the most important aspects of Acanthamoeba biology and their interactions with endemically important human pathogens.

  13. Isolation and genotyping of Acanthamoeba strains from corneal infections in Italy.

    PubMed

    Gatti, Simonetta; Rama, Paolo; Matuska, Stanislav; Berrilli, Federica; Cavallero, Annalisa; Carletti, Silvia; Bruno, Antonella; Maserati, Roberta; Di Cave, David

    2010-11-01

    Acanthamoeba keratitis (AK) is a corneal disease caused by members of a genus of free-living amoebae and is associated predominantly with contact lens (CL) use. This study reports 16 cases of culture-proven AK diagnosed in northern Italy. Genotype identification was carried out with a PCR assay based on sequence analysis of the 18S rRNA gene, and sensitivity and specificity were evaluated in comparison with traditional parasitological techniques. A 405 bp region of the 18S rRNA gene (ASA.S1) including diagnostic fragment 3 (DF3) was amplified using the genus-specific primers JDP1 and JDP2. Genotype assignment was based on phenetic analysis of the ASA.S1 subset of the nuclear small-subunit rRNA gene sequence excluding the highly variable DF3 region. Phylogenetic analysis was also performed on the sequences obtained. All patients complained of monolateral infection; 11 (68.75%) admitted improper CL disinfection. In 14/16 (87.5 %) subjects, corneal scrapings were stained with calcofluor white and haematoxylin and eosin and, in ten cases (62.5 %), microscopy was positive for Acanthamoeba cysts. In vitro culture on 3 % non-nutrient agar plates was obtained in all cases (100 %), whereas cloning and axenic growth were positive for 14 amoebic stocks (87.5 %). PCR analysis had 100 % sensitivity and specificity compared with in vitro axenic culture, showing positive amplification from 15 isolates. All Acanthamoeba strains belonged to the T4 genotype, the main AK-related genotype worldwide. These results confirmed the importance of a complete diagnostic protocol, including a PCR assay, for the clinical diagnosis of AK on biological samples. Genotyping allowed inclusion of all isolates in the T4 group, thus demonstrating the prevalence of this genotype in northern Italy.

  14. Acanthamoeba encystment: multifactorial effects of buffers, biocides, and demulcents present in contact lens care solutions

    PubMed Central

    Kovacs, Christopher J; Lynch, Shawn C; Rah, Marjorie J; Millard, Kimberly A; Morris, Timothy W

    2015-01-01

    Purpose To determine whether agents which are purportedly capable of inducing encystment of Acanthamoeba can recapitulate the signal when tested in differing formulations. Methods In accordance with the International Standard ISO 19045, Acanthamoeba castellanii ATCC 50370 trophozoites were cultured in antibiotic-free axenic medium, treated with test solutions, and encystment rates plus viability were measured via bright field and fluorescent microscopy. Test solutions included phosphate-buffered saline (PBS), borate-buffered saline, biguanide- and hydrogen peroxide (H2O2)-based biocides, propylene glycol (PG) and povidone (POV) ophthalmic demulcents, and one-step H2O2-based contact lens disinfection systems. Results Only PBS solutions with 0.25 ppm polyaminopropyl biguanide (PAPB) and increasing concentrations of PG and POV stimulated A. castellanii encystment in a dose-dependent manner, whereas PBS solutions containing 3% H2O2 and increasing concentrations of PG and POV did not stimulate encystment. Borate-buffered saline and PBS/citrate solutions containing PG also did not stimulate encystment. In addition, no encystment was observed after 24 hours, 7 days, or 14 days of exposures of trophozoites to one-step H2O2 contact lens disinfection products or related solutions. Conclusion The lack of any encystment observed when trophozoites were treated with existing or new one-step H2O2 contact lens care products, as well as when trophozoites were exposed to various related test solutions, confirms that Acanthamoeba encystment is a complex process which depends upon simultaneous contributions of multiple factors including buffers, biocides, and demulcents. PMID:26508829

  15. Comparison of Hydrogen Peroxide Contact Lens Disinfection Systems and Solutions against Acanthamoeba polyphaga

    PubMed Central

    Hughes, Reanne; Kilvington, Simon

    2001-01-01

    Acanthamoeba is a free-living amoeba causing a potentially blinding infection of the cornea. Contact lens wearers are most at risk and account for some 95% of cases. Hydrogen peroxide is used for contact lens disinfection due to its broad antimicrobial activity. Lenses must be neutralized before use to avoid pronounced stinging and possible corneal damage. Neutralization is achieved by adding a catalyst during the disinfection process (one-step) or afterwards (two-step). Here, the activities of commercial peroxide systems and individual solutions against trophozoites and cysts of Acanthamoeba polyphaga were compared. All disinfection systems were active against trophozoites, giving a ≥3-log (99.9%) kill within 1 h. Of the four one-step systems, only one showed some cysticidal activity, giving a 1.28 ± 0.41-log reduction. Both two-step systems were cysticidal, giving a ≥3-log kill at 4 h. All system peroxide solutions were cysticidal, giving a ≥3-log kill by 4 to 6 h. Variation in the cysticidal rate was observed with two solutions that gave a 1.8- to 2.1-log kill at 4 h compared with 3.0 to 4.0 for the rest (P < 0.05). No cysticidal activity was found with the peroxigen sodium perborate or the contact lens protein remover subtilisin A. Two-step systems are cysticidal providing contact times of at least 4 h are employed. Variation in cyst killing occurs between peroxide solutions, possibly due to formulation differences. One-step systems are less effective against Acanthamoeba cysts due to rapid peroxide neutralization. The cysticidal activity of one-step systems could be improved if neutralization rates were retarded. PMID:11408220

  16. Enhanced killing of Acanthamoeba cysts with a plant peroxidase-hydrogen peroxide-halide antimicrobial system.

    PubMed

    Hughes, Reanne; Andrew, Peter W; Kilvington, Simon

    2003-05-01

    The activity of H(2)O(2) against the resistant cyst stage of the pathogenic free-living amoeba Acanthamoeba was enhanced by the addition of KI and either horseradish peroxidase or soybean peroxidase or, to a lesser degree, lactoperoxidase. This resulted in an increase in the cysticidal activity of 3% (wt/vol) H(2)O(2), and there was >3-log killing in 2 h, compared with the 6 h required for comparable results with the peroxide solution alone (P < 0.05). With 2% H(2)O(2), enhancement was observed at all time points (P < 0.05), and total killing of the cyst inoculum occurred at 4 h, compared with 6 h for the peroxide alone. The activity of sublethal 1% H(2)O(2) was enhanced to give 3-log killing after 8 h of exposure (P < 0.05). No enhancement was obtained when KCl or catalase was used as a substitute in the reaction mixtures. The H(2)O(2) was not neutralized in the enhanced system during the experiments. However, in the presence of a platinum disk used to neutralize H(2)O(2) in contact lens care systems, the enhanced 2% H(2)O(2) system gave 2.8-log killing after 6 h or total cyst killing by 8 h, and total neutralization of the H(2)O(2) occurred by 4 h. In contrast, 2% H(2)O(2) alone resulted in <0.8-log killing of cysts in the presence of the platinum disk due to rapid (<1 h) neutralization of the peroxide. Our observations could result in significant improvement in the efficacy of H(2)O(2) contact lens disinfection systems against Acanthamoeba cysts and prevention of acanthamoeba keratitis.

  17. Enhanced Killing of Acanthamoeba Cysts with a Plant Peroxidase-Hydrogen Peroxide-Halide Antimicrobial System

    PubMed Central

    Hughes, Reanne; Andrew, Peter W.; Kilvington, Simon

    2003-01-01

    The activity of H2O2 against the resistant cyst stage of the pathogenic free-living amoeba Acanthamoeba was enhanced by the addition of KI and either horseradish peroxidase or soybean peroxidase or, to a lesser degree, lactoperoxidase. This resulted in an increase in the cysticidal activity of 3% (wt/vol) H2O2, and there was >3-log killing in 2 h, compared with the 6 h required for comparable results with the peroxide solution alone (P < 0.05). With 2% H2O2, enhancement was observed at all time points (P < 0.05), and total killing of the cyst inoculum occurred at 4 h, compared with 6 h for the peroxide alone. The activity of sublethal 1% H2O2 was enhanced to give 3-log killing after 8 h of exposure (P < 0.05). No enhancement was obtained when KCl or catalase was used as a substitute in the reaction mixtures. The H2O2 was not neutralized in the enhanced system during the experiments. However, in the presence of a platinum disk used to neutralize H2O2 in contact lens care systems, the enhanced 2% H2O2 system gave 2.8-log killing after 6 h or total cyst killing by 8 h, and total neutralization of the H2O2 occurred by 4 h. In contrast, 2% H2O2 alone resulted in <0.8-log killing of cysts in the presence of the platinum disk due to rapid (<1 h) neutralization of the peroxide. Our observations could result in significant improvement in the efficacy of H2O2 contact lens disinfection systems against Acanthamoeba cysts and prevention of acanthamoeba keratitis. PMID:12732522

  18. Evaluation of inhibitory potential of some selective methanolic plants extracts on biological characteristics of Acanthamoeba castellanii using human corneal epithelial cells in vitro.

    PubMed

    Shoaib, Hafiz Muhammad; Muazzam, Ambreen Gul; Mir, Asif; Jung, Suk-Yul; Matin, Abdul

    2013-03-01

    Acanthamoeba is an opportunistic protozoan pathogen and known to be one of the most ubiquitous organisms, play a vital role in ecosystem, and recognized to cause blinding keratitis and rare but fatal granulomatous encephalitis involving the central nervous system with a very poor prognosis. This is due to limited availability of effective anti-Acanthamoeba drugs. The objective of the present study was to determine the efficacy of methanolic plants crude extracts on the viability and biological properties of Acanthamoeba castellanii (T4 genotype) and its cytotoxic effects on human corneal epithelial cells (HCEC). Using HCEC, it was observed that Acanthamoeba exhibited binding (>90 %) and cytotoxicity (>80 %) to host cells. However, plant crude extracts remarkably inhibited more than 70 and 60 % of Acanthamoeba binding and cytotoxicity to HCEC, respectively. It was further established that crude extracts (ranging from 0.1 to 1.5 mg/ml) exhibited amoebicidal effects, i.e., >50 % of trophozoites were killed/reduced at maximum dose (1.5 mg/ml) within 1 h incubation. However, the residual subpopulation remained static over longer incubations. Furthermore, growth assay demonstrated crude extracts inhibited >50 % Acanthamoeba numbers up to 7 days. Our results confirmed that plant crude extracts has inhibitory effects on Acanthamoeba growth and viability. Overall, these findings revealed that tested plant extracts is inhibitory to Acanthamoeba properties associated with pathogenesis. To the best of our knowledge, our findings demonstrated for the first time that selected methanol plant crude extracts exhibits inhibitory effects on biological properties of Acanthamoeba without any toxic effects on HCEC cells in vitro.

  19. Detection of free living amoebae, Acanthamoeba and Naegleria, in swimming pools, Malaysia.

    PubMed

    Init, I; Lau, Y L; Arin Fadzlun, A; Foead, A I; Neilson, R S; Nissapatorn, V

    2010-12-01

    This study reports the detection of Acanthamoeba and Naegleria species in 14 swimming pools around Petaling Jaya and Kuala Lumpur, Malaysia. Sampling was carried out at 4 sites (the platforms (P), wall (W), 1 meter from the wall (1) and middle (2)) of each swimming pool. These free living amoebae (FLA) were detected under light and inverted microscopes after being cultured on the surface of non-nutrient agar lawned with Escherichia coli. Acanthamoeba species were detected in higher number of culture plates from all sampling sites of all the swimming pools. While Naegleria, were detected in fewer culture plates at 3 sampling sites (absent at site P) of 8 swimming pools. This suggested that the thick double-walled cysts of Acanthamoeba were more resistant, thus remaining viable in the dry-hot areas of the platforms and in chlorinated water of the swimming pools whereas Naegleria cysts, that are fragile and susceptible to desiccation, preferred watery or moist areas for growth and proliferation. The prevalence of both FLA was highest at site W (76.2%), followed by site 1 (64.7%), lowest at site 2 (19.4%), and could be detected at all 3 sampling levels (top, middle and bottom) of these 3 sites. The surface of site W might act as a bio-film that accumulated all kinds of microbes providing sufficient requirement for the FLA to develop and undergo many rounds of life cycles as well as moving from top to bottom in order to graze food. Other factors such as human activities, the circulating system which was fixed at all swimming pools, blowing wind which might carry the cysts from surroundings and the swimming flagellate stage of Naegleria could also contribute to the distribution of the FLA at these sampling sites. Both FLA showed highest growth (80.4%) at room temperature (25-28 ºC) and lesser (70.0%) at 37 ºC which might be due to the overgrowth of other microbes (E. coli, fungi, algae, etc). While at 44 ºC, only Acanthamoeba species could survive thus showing that

  20. A novel antiamoebic agent against Acanthamoeba sp. - A causative agent for eye keratitis infection

    NASA Astrophysics Data System (ADS)

    Kusrini, Eny; Hashim, Fatimah; Azmi, Wan Nor Nadhirah Wan Noor; Amin, Nakisah Mat; Estuningtyas, Ari

    2016-01-01

    The terbium trinitrate.trihydrate.18-crown ether-6, Tb(NO3)3(OH2)3.(18C6) complex has been characterized by elemental analysis, photoluminescence and single X-ray diffraction. The IC50 values were determined based on MTT assay while light and fluorescence microscopy imaging were employed to evaluate the cellular morphological changes. Alkaline comet assay was performed to analyze the DNA damage. The photoluminescence spectrum of the Tb complex excited at 325 nm displayed seven luminescence peaks corresponding to the 5D4 → 7F0, 1, 2, 3, 4, 5, 6 transitions. The cytotoxicity and genotoxicity studies indicated that the Tb(NO3)3(OH2)3.(18C6) complex and its salt form as well as the 18C6 molecule have excellent anti-amoebic activity with very low IC50 values are 7, 2.6 and 1.2 μg/mL, respectively, with significant decrease (p < 0.05) in Acanthamoeba viability when the concentration was increased from 0 to 30 μg/mL. The mode of cell death in Acanthamoeba cells following treatment with the Tb complex was apoptosis. This is in contrast to the Tb(NO3)3.6H2O salt- and 18C6 molecule-treated Acanthamoeba, which exhibited necrotic type cells. The percentage of DNA damage following treatment with all the compounds at the IC25 values showed high percentage of type 1 with the % nuclei damage are 14.15 ± 2.4; 46.00 ± 4.2; 36.36 ± 2.4; 45.16 ± 0.6%, respectively for untreated, treated with Tb complex, Tb salt and 18C6 molecule. The work features promising potential of Tb(NO3)3(OH2)3.(18C6) complex as anti-amoebic agent, representing a therapeutic option for Acanthamoeba keratitis infection.

  1. A novel antiamoebic agent against Acanthamoeba sp.--A causative agent for eye keratitis infection.

    PubMed

    Kusrini, Eny; Hashim, Fatimah; Azmi, Wan Nor Nadhirah Wan Noor; Amin, Nakisah Mat; Estuningtyas, Ari

    2016-01-15

    The terbium trinitrate.trihydrate.18-crown ether-6, Tb(NO3)3(OH2)3.(18C6) complex has been characterized by elemental analysis, photoluminescence and single X-ray diffraction. The IC50 values were determined based on MTT assay while light and fluorescence microscopy imaging were employed to evaluate the cellular morphological changes. Alkaline comet assay was performed to analyze the DNA damage. The photoluminescence spectrum of the Tb complex excited at 325 nm displayed seven luminescence peaks corresponding to the (5)D4→(7)F(0, 1, 2, 3, 4, 5, 6) transitions. The cytotoxicity and genotoxicity studies indicated that the Tb(NO3)3(OH2)3.(18C6) complex and its salt form as well as the 18C6 molecule have excellent anti-amoebic activity with very low IC50 values are 7, 2.6 and 1.2 μg/mL, respectively, with significant decrease (p<0.05) in Acanthamoeba viability when the concentration was increased from 0 to 30 μg/mL. The mode of cell death in Acanthamoeba cells following treatment with the Tb complex was apoptosis. This is in contrast to the Tb(NO3)3.6H2O salt- and 18C6 molecule-treated Acanthamoeba, which exhibited necrotic type cells. The percentage of DNA damage following treatment with all the compounds at the IC25 values showed high percentage of type 1 with the % nuclei damage are 14.15±2.4; 46.00±4.2; 36.36±2.4; 45.16±0.6%, respectively for untreated, treated with Tb complex, Tb salt and 18C6 molecule. The work features promising potential of Tb(NO3)3(OH2)3.(18C6) complex as anti-amoebic agent, representing a therapeutic option for Acanthamoeba keratitis infection.

  2. Acanthamoeba polyphaga mimivirus and other giant viruses: an open field to outstanding discoveries

    PubMed Central

    2014-01-01

    In 2003, Acanthamoeba polyphaga mimivirus (APMV) was first described and began to impact researchers around the world, due to its structural and genetic complexity. This virus founded the family Mimiviridae. In recent years, several new giant viruses have been isolated from different environments and specimens. Giant virus research is in its initial phase and information that may arise in the coming years may change current conceptions of life, diversity and evolution. Thus, this review aims to condense the studies conducted so far about the features and peculiarities of APMV, from its discovery to its clinical relevance. PMID:24976356

  3. Acanthamoeba polyphaga mimivirus and other giant viruses: an open field to outstanding discoveries.

    PubMed

    Abrahão, Jônatas S; Dornas, Fábio P; Silva, Lorena C F; Almeida, Gabriel M; Boratto, Paulo V M; Colson, Phillipe; La Scola, Bernard; Kroon, Erna G

    2014-06-30

    In 2003, Acanthamoeba polyphaga mimivirus (APMV) was first described and began to impact researchers around the world, due to its structural and genetic complexity. This virus founded the family Mimiviridae. In recent years, several new giant viruses have been isolated from different environments and specimens. Giant virus research is in its initial phase and information that may arise in the coming years may change current conceptions of life, diversity and evolution. Thus, this review aims to condense the studies conducted so far about the features and peculiarities of APMV, from its discovery to its clinical relevance.

  4. Genome-wide association study for refractive astigmatism reveals genetic co-determination with spherical equivalent refractive error: the CREAM consortium.

    PubMed

    Li, Qing; Wojciechowski, Robert; Simpson, Claire L; Hysi, Pirro G; Verhoeven, Virginie J M; Ikram, Mohammad Kamran; Höhn, René; Vitart, Veronique; Hewitt, Alex W; Oexle, Konrad; Mäkelä, Kari-Matti; MacGregor, Stuart; Pirastu, Mario; Fan, Qiao; Cheng, Ching-Yu; St Pourcain, Beaté; McMahon, George; Kemp, John P; Northstone, Kate; Rahi, Jugnoo S; Cumberland, Phillippa M; Martin, Nicholas G; Sanfilippo, Paul G; Lu, Yi; Wang, Ya Xing; Hayward, Caroline; Polašek, Ozren; Campbell, Harry; Bencic, Goran; Wright, Alan F; Wedenoja, Juho; Zeller, Tanja; Schillert, Arne; Mirshahi, Alireza; Lackner, Karl; Yip, Shea Ping; Yap, Maurice K H; Ried, Janina S; Gieger, Christian; Murgia, Federico; Wilson, James F; Fleck, Brian; Yazar, Seyhan; Vingerling, Johannes R; Hofman, Albert; Uitterlinden, André; Rivadeneira, Fernando; Amin, Najaf; Karssen, Lennart; Oostra, Ben A; Zhou, Xin; Teo, Yik-Ying; Tai, E Shyong; Vithana, Eranga; Barathi, Veluchamy; Zheng, Yingfeng; Siantar, Rosalynn Grace; Neelam, Kumari; Shin, Youchan; Lam, Janice; Yonova-Doing, Ekaterina; Venturini, Cristina; Hosseini, S Mohsen; Wong, Hoi-Suen; Lehtimäki, Terho; Kähönen, Mika; Raitakari, Olli; Timpson, Nicholas J; Evans, David M; Khor, Chiea-Chuen; Aung, Tin; Young, Terri L; Mitchell, Paul; Klein, Barbara; van Duijn, Cornelia M; Meitinger, Thomas; Jonas, Jost B; Baird, Paul N; Mackey, David A; Wong, Tien Yin; Saw, Seang-Mei; Pärssinen, Olavi; Stambolian, Dwight; Hammond, Christopher J; Klaver, Caroline C W; Williams, Cathy; Paterson, Andrew D; Bailey-Wilson, Joan E; Guggenheim, Jeremy A

    2015-02-01

    To identify genetic variants associated with refractive astigmatism in the general population, meta-analyses of genome-wide association studies were performed for: White Europeans aged at least 25 years (20 cohorts, N = 31,968); Asian subjects aged at least 25 years (7 cohorts, N = 9,295); White Europeans aged <25 years (4 cohorts, N = 5,640); and all independent individuals from the above three samples combined with a sample of Chinese subjects aged <25 years (N = 45,931). Participants were classified as cases with refractive astigmatism if the average cylinder power in their two eyes was at least 1.00 diopter and as controls otherwise. Genome-wide association analysis was carried out for each cohort separately using logistic regression. Meta-analysis was conducted using a fixed effects model. In the older European group the most strongly associated marker was downstream of the neurexin-1 (NRXN1) gene (rs1401327, P = 3.92E-8). No other region reached genome-wide significance, and association signals were lower for the younger European group and Asian group. In the meta-analysis of all cohorts, no marker reached genome-wide significance: The most strongly associated regions were, NRXN1 (rs1401327, P = 2.93E-07), TOX (rs7823467, P = 3.47E-07) and LINC00340 (rs12212674, P = 1.49E-06). For 34 markers identified in prior GWAS for spherical equivalent refractive error, the beta coefficients for genotype versus spherical equivalent, and genotype versus refractive astigmatism, were highly correlated (r = -0.59, P = 2.10E-04). This work revealed no consistent or strong genetic signals for refractive astigmatism; however, the TOX gene region previously identified in GWAS for spherical equivalent refractive error was the second most strongly associated region. Analysis of additional markers provided evidence supporting widespread genetic co-susceptibility for spherical and astigmatic refractive errors.

  5. Hard x-ray spectroscopy and imaging by a reflection zone plate in the presence of astigmatism

    SciTech Connect

    Braig, Christoph; Lochel, Heike; Firsov, Alexander; Brzhezinskaya, Maria; Hafner, Aljosa; Rehanek, Jens; Wojcik, Michael; Macrander, Albert; Assoufid, Lahsen; Erko, Alexei

    2015-12-17

    Here, the feasibility of an off-axis x-ray reflection zone plate to perform wavelength-dispersive spectroscopy, on-axis point focusing, and two-dimensional imaging is demonstrated by means of one and the same diffractive optical element (DOE) at a synchrotron radiation facility. The resolving power varies between 3 × 101 and 4 × 102 in the range of 7.6 keV to 9.0 keV, with its maximum at the design energy of 8.3 keV. This result is verified using an adjustable entrance slit, by which horizontal (H) and vertical (V) focusing to 0.85 μm(H) and 1.29 μm(V) is obtained near the sagittal focal plane of the astigmatic configuration. An angular and axial scan proves an accessible field of view of at least 0.6 arcmin × 0.8 arcmin and a focal depth of ±0.86 mm. Supported by the grating efficiency of around 17.5% and a very short pulse elongation, future precision x-ray fluorescence and absorption studies of transition metals at their K-edge on an ultrashort timescale could benefit from our findings.

  6. Hard x-ray spectroscopy and imaging by a reflection zone plate in the presence of astigmatism

    DOE PAGES

    Braig, Christoph; Lochel, Heike; Firsov, Alexander; ...

    2015-12-17

    Here, the feasibility of an off-axis x-ray reflection zone plate to perform wavelength-dispersive spectroscopy, on-axis point focusing, and two-dimensional imaging is demonstrated by means of one and the same diffractive optical element (DOE) at a synchrotron radiation facility. The resolving power varies between 3 × 101 and 4 × 102 in the range of 7.6 keV to 9.0 keV, with its maximum at the design energy of 8.3 keV. This result is verified using an adjustable entrance slit, by which horizontal (H) and vertical (V) focusing to 0.85 μm(H) and 1.29 μm(V) is obtained near the sagittal focal plane ofmore » the astigmatic configuration. An angular and axial scan proves an accessible field of view of at least 0.6 arcmin × 0.8 arcmin and a focal depth of ±0.86 mm. Supported by the grating efficiency of around 17.5% and a very short pulse elongation, future precision x-ray fluorescence and absorption studies of transition metals at their K-edge on an ultrashort timescale could benefit from our findings.« less

  7. Severe Amoebic Placentitis in a Horse Caused by an Acanthamoeba hatchetti Isolate Identified Using Next-Generation Sequencing

    PubMed Central

    Begg, Angela P.; Todhunter, Kristen; Donahoe, Shannon L.; Krockenberger, Mark

    2014-01-01

    A case of amoebic placentitis in a mare from eastern Australia was diagnosed postpartum by histopathological examination of the placenta. The identity of the etiological agent was confirmed as Acanthamoeba hatchetti by use of diversity profiling based on a next-generation sequencing approach. PMID:24829227

  8. The affinities of human platelet and Acanthamoeba profilin isoforms for polyphosphoinositides account for their relative abilities to inhibit phospholipase C.

    PubMed Central

    Machesky, L M; Goldschmidt-Clermont, P J; Pollard, T D

    1990-01-01

    In light of recent work implicating profilin from human platelets as a possible regulator of both cytoskeletal dynamics and inositol phospholipid-mediated signaling, we have further characterized the interaction of platelet profilin and the two isoforms of Acanthamoeba profilin with inositol phospholipids. Profilin from human platelets binds to phosphatidylinositol-4-monophosphate (PIP) and phosphatidylinositol-4,5-bisphosphate (PIP2) with relatively high affinity (Kd approximately 1 microM for PIP2 by equilibrium gel filtration), but interacts only weakly (if at all) with phosphatidylinositol (PI) or inositol trisphosphate IP3) in small-zone gel-filtration assays. The two isoforms of Acanthamoeba profilin both have a lower affinity for PIP2 than does human platelet profilin, but the more basic profilin isoform from Acanthamoeba (profilin-II) has a much higher (approximately 10-microM Kd) affinity than the acidic isoform (profilin-I, 100 to 500-microM Kd). None of the profilins bind to phosphatidylserine (PS) or phosphatidylcholine (PC) in small-zone gel-filtration experiments. The differences in affinity for PIP2 parallel the ability of these three profilins to inhibit PIP2 hydrolysis by soluble phospholipase C (PLC). The results show that the interaction of profilins with PIP2 is specific with respect to both the lipid and the proteins. In Acanthamoeba, the two isoforms of profilin may have specialized functions on the basis of their identical (approximately 10 microM) affinities for actin monomers and different affinities for PIP2. PMID:1966040

  9. Phylogenetic analysis and the evolution of the 18S rRNA gene typing system of Acanthamoeba.

    PubMed

    Fuerst, Paul A; Booton, Gregory C; Crary, Monica

    2015-01-01

    Species of Acanthamoeba were first described using morphological characters including cyst structure and cytology of nuclear division. More than 20 nominal species were proposed using these methods. Morphology, especially cyst shape and size, has proven to be plastic and dependent upon culture conditions. The DNA sequence of the nuclear small-subunit (18S) rRNA, the Rns gene, has become the most widely accepted method for rapid diagnosis and classification of Acanthamoeba. The Byers-Fuerst lab first proposed an Rns typing system in 1996. Subsequent refinements, with an increasing DNA database and analysis of diagnostic fragments within the gene, have become widely accepted by the Acanthamoeba research community. The development of the typing system, including its current state of implementation is illustrated by three cases: (i) the division between sequence types T13 and T16; (ii) the diversity within sequence supertype T2/T6, and (iii) verification of a new sequence type, designated T20. Molecular studies make clear the disconnection between phylogenetic relatedness and species names, as applied for the genus Acanthamoeba. Future reconciliation of genetic types with species names must become a priority, but the possible shortcomings of the use of a single gene when reconstructing the evolutionary history of the acanthamoebidae must also be resolved.

  10. Monitoring of in vitro dynamics of Acanthamoeba strains isolated from infected eyes as a useful tool in keratitis management.

    PubMed

    Chomicz, Lidia; Padzik, Marcin; Szaflik, Jacek P; Nahorski, Wacław L; Kryczka, Tomasz; Szaflik, Jerzy

    2014-11-01

    Free-living amoebae of Acanthamoeba genus are ubiquitous in various parts of the world. Some species of these amoebozoans present a serious risk to human health as the causative agents of vision-threatening diseases, Acanthamoeba keratitis. Correct diagnosis requires both a clinical examination of the cornea and amoebic form identification in affected eyes. Despite advances in pharmacotherapy, the infection is difficult to diagnose and to threat. Population dynamics of five different Acanthamoeba strains cultured in vitro under bacteria-free condition in BSC medium, was monitored in terms of diagnostic and therapeutic management. The range of protozoan number in the exponential growth phase, the morpho-physiological status of amoeba forms and their ability to multiply were evaluated. Results of the studies revealed that early and continued monitoring of the strains maintained in an axenic culture showed correlation between the dynamics of cultivated amoebae and the course of the disease, differences in response to pharmacotherapy and the surgical management efficacy. Concluding, the in vitro monitoring of dynamics of Acanthamoeba strains isolated from infected corneas may be important not only for proper diagnosis but also as a useful tool in keratitis management and therapeutic prognosis.

  11. Acanthamoeba belonging to T3, T4, and T11: genotypes isolated from air-conditioning units in Santiago, Chile.

    PubMed

    Astorga, Berbeli; Lorenzo-Morales, Jacob; Martín-Navarro, Carmen M; Alarcón, Verónica; Moreno, Johanna; González, Ana C; Navarrete, Elizabeth; Piñero, José E; Valladares, Basilio

    2011-01-01

    Free-living amoebae (FLA) of the genus Acanthamoeba are widely distributed in the environment, in the air, soil, and water, and have also been isolated from air-conditioning units. The objective of this work was to investigate the presence of this genus of FLA in the air-conditioning equipment at the Institute of Public Health of Chile in Santiago, Chile. Water and air samples were collected from air-conditioning systems and were checked for the presence of Acanthamoeba spp. Positive samples were further classified at the genotype level after sequencing the highly variable diagnostic fragment 3 (DF3) region of the 18S rRNA gene. This is the first report of the T3, T4, and T11 genotypes of Acanthamoeba in air-conditioning units from Chile. Overall, the widespread distribution of potentially pathogenic Acanthamoeba strains in the studied source demands more awareness within the public and health professionals in Chile as this pathogen is emerging as a risk for human health worldwide.

  12. Identification of a novel t17 genotype of acanthamoeba from environmental isolates and t10 genotype causing keratitis in Thailand.

    PubMed

    Nuprasert, Warisa; Putaporntip, Chaturong; Pariyakanok, Lalida; Jongwutiwes, Somchai

    2010-12-01

    We analyzed the nuclear small-subunit rRNA genes of Acanthamoeba isolates from freshwater sources (n=16) and from patients (n=6) in Bangkok and surrounding areas. The T10 genotype from a keratitis patient and a novel T17 genotype from water samples were diagnosed for the first time in this study.

  13. Viability of Acanthamoeba after exposure to a multipurpose disinfecting contact lens solution and two hydrogen peroxide systems

    PubMed Central

    Hiti, K; Walochnik, J; Haller-Schober, E M; Faschinger, C; Aspöck, H

    2002-01-01

    Background/aim: Contact lens cases contaminated with Acanthamoeba are a major risk factor for an infection of the eye. In this study the anti-Acanthamoeba activity of three different contact lens storage solutions was tested. Methods: A new multipurpose contact lens storage solution (Meni Care Plus) and a two step (Titmus H2O2) and one step (Oxysept Comfort) hydrogen peroxide system were tested for their effects on trophozoites and cysts of three different Acanthamoeba species: A castellanii, A hatchetti, and A lenticulata. Results: After a soaking time of 8 hours (overnight soaking of contact lenses) the Titmus H2O2 0.6% solution showed very good amoebicidal effects, while Oxysept Comfort 3% H2O2 could not effectively destroy the cysts of any of the three tested species. Viable cysts of the species A lenticulata and A hatchetti were still present after exposure to Meni Care Plus (0.0005% PHMB) for 8 hours. Conclusion: Not all of the three tested contact lens storage solutions have sufficient amoebicidal effects. The two step peroxide system Titmus H2O2 is a very effective disinfectant contact lens solution in order to avoid a possible Acanthamoeba infection of the eye. PMID:11815336

  14. How Could Contact Lens Wearers Be at Risk of Acanthamoeba Infection? A Review

    PubMed Central

    Ibrahim, Youhanna W.; Boase, David L.; Cree, Ian A.

    2010-01-01

    Contact lens wear is highly influential on the incidence of ulcerative keratitis worldwide, particularly in developed countries. The association between Acanthamoeba keratitis and contact lens wear is firmly established; it may account for up to 95% of the reported cases. Before the popularisation of soft contact lens wear, Acanthamoeba keratitis was extremely rare. In 2000 it was estimated that the number of contact lens wearers worldwide was about 80 million, out of whom 33 million were in the United States and 90% of them wore hydrogel soft lenses. Contact lens-related problems depend on many factors, such as lens material, wearing modality, lens hygiene, type of lens-caring solution, the degree of compliance of the lens user with lens wear and care procedures, lens overwear, sleeping in lenses, rate of changing lenses, and lens case hygiene. This paper is a thorough review of the literature aiming to highlight the role of one of the main risk factors of infectious keratitis, contact lens wear, and also to show the responsibility of lens users in aggravating this risk.

  15. Campylobacter jejuni Actively Invades the Amoeba Acanthamoeba polyphaga and Survives within Non Digestive Vacuoles

    PubMed Central

    Olofsson, Jenny; Axelsson-Olsson, Diana; Brudin, Lars; Olsen, Björn; Ellström, Patrik

    2013-01-01

    The Gram-negative bacterium Campylobacter jejuni is able to enter, survive and multiply within the free living amoeba Acanthamoeba polyphaga, but the molecular mechanisms behind these events are still unclear. We have studied the uptake and intracellular trafficking of viable and heat killed bacterial cells of the C. jejuni strain 81–176 in A. polyphaga. We found that viable bacteria associated with a substantially higher proportion of Acanthamoeba trophozoites than heat killed bacteria. Furthermore, the kinetics of internalization, the total number of internalized bacteria as well as the intracellular localization of internalized C. jejuni were dramatically influenced by bacterial viability. Viable bacteria were internalized at a high rate already after 1 h of co-incubation and were observed in small vacuoles tightly surrounding the bacteria. In contrast, internalization of heat killed C. jejuni was low at early time points and did not peak until 96 h. These cells were gathered in large spacious vacuoles that were part of the degradative pathway as determined by the uptake of fluorescently labeled dextran. The amount of heat killed bacteria internalized by A. polyphaga did never reach the maximal amount of internalized viable bacteria. These results suggest that the uptake and intracellular survival of C. jejuni in A. polyphaga is bacterially induced. PMID:24223169

  16. Behavior of Yersinia enterocolitica in the Presence of the Bacterivorous Acanthamoeba castellanii

    PubMed Central

    Lambrecht, E.; Baré, J.; Van Damme, I.; Bert, W.; Sabbe, K.

    2013-01-01

    Free-living protozoa play an important role in the ecology and epidemiology of human-pathogenic bacteria. In the present study, the interaction between Yersinia enterocolitica, an important food-borne pathogen, and the free-living amoeba Acanthamoeba castellanii was studied. Several cocultivation assays were set up to assess the resistance of Y. enterocolitica to A. castellanii predation and the impact of environmental factors and bacterial strain-specific characteristics. Results showed that all Y. enterocolitica strains persist in association with A. castellanii for at least 14 days, and associations with A. castellanii enhanced survival of Yersinia under nutrient-rich conditions at 25°C and under nutrient-poor conditions at 37°C. Amoebae cultivated in the supernatant of one Yersinia strain showed temperature- and time-dependent permeabilization. Intraprotozoan survival of Y. enterocolitica depended on nutrient availability and temperature, with up to 2.8 log CFU/ml bacteria displaying intracellular survival at 7°C for at least 4 days in nutrient-rich medium. Transmission electron microscopy was performed to locate the Yersinia cells inside the amoebae. As Yersinia and Acanthamoeba share similar ecological niches, this interaction identifies a role of free-living protozoa in the ecology and epidemiology of Y. enterocolitica. PMID:23934496

  17. Investigating the role of Acanthamoeba polyphaga in protecting Human Adenovirus from water disinfection treatment.

    PubMed

    Verani, Marco; Di Giuseppe, Graziano; Tammaro, Carmine; Carducci, Annalaura

    2016-06-01

    Human adenoviruses are responsible for a wide range of clinical infections and are present in aquatic environments, including river, seawater, drinking-water and sewage. Free-living amoebae (Acanthamoeba) in the same environments may internalize them and other microorganisms can act as a reservoir for the internalized viruses. In this study, we studied the interaction between Acanthamoeba polyphaga and Human Adenovirus type 5 (HAdV 5) to determine whether the amoeba played a role in protecting the internalized viruses from chemical disinfection. The efficacy of sodium hypochlorite disinfection against A. polyphaga and HAdV 5 either singly or in combination was assessed at three different concentrations. Individually, the amoeba were more resistant to chemical disinfection than HAdV 5 and remained alive after exposure to 5mg/l of sodium hypochlorite. In contrast, HAdV 5 lost infectivity following exposure to 2.5mg/l of sodium hypochlorite. When the amoeba and HAdV 5 were co-cultured, infectious virus was found in the cytoplasm of the amoeba at 5mg/l disinfectant concentration. These findings suggest that the A. polyphaga is providing protection for the HAdV 5.

  18. Temperature limitation may explain the containment of the trophozoites in the cornea during Acanthamoeba castellanii keratitis.

    PubMed

    Nielsen, Mattias Kiel; Nielsen, Kim; Hjortdal, Jesper; Sørensen, Uffe B Skov

    2014-12-01

    Acanthamoeba keratitis is a serious sight-threatening disease. The relatively low temperature of the cornea may explain why amoebic infections usually are localized in this tissue and rarely spread to other parts of the eye. In this study, the growth rate of the amoeba Acanthamoeba castellanii was examined at different temperatures. The aim was to establish the optimal growth temperature for A. castellanii and to examine the growth within the vicinity of the core body temperature. The growth rates of four clinical and two environmental strains of A. castellanii were estimated at different temperatures, and temperature limitations for the trophozoite stage was established. Movements influenced by temperature gradients were monitored for two clinical strains of A. castellanii. The highest growth rate for each of the six amoebic strains tested was found to be close to 32 °C. The growth of the trophozoites of all examined strains was greatly reduced or completely halted at temperatures above 36 °C and encysted at the elevated temperature. Thus, the optimal growth temperature for the four strains of A. castellanii is close to the surface temperature of the human cornea, while the higher body core-temperature induced encysting of the amoebae. This may explain why most amoebic eye infections are confined to the cornea.

  19. Influence of Legionella pneumophila and other water bacteria on the survival and growth of Acanthamoeba polyphaga.

    PubMed

    Anacarso, I; Guerrieri, E; Bondi, M; de Niederhäusern, S; Iseppi, R; Sabia, C; Contri, M; Borella, P; Messi, P

    2010-10-01

    We investigated in solid medium, in water microcosm co-cultures and by light and transmission electron microscopy the influence of Legionella pneumophila Lp-1, Pseudomonas aeruginosa ATCC 27853, Burkholderia cepacia ATCC 25416 and Pseudomonas fluorescens SSD35 on the growth and survival of Acanthamoeba polyphaga. The infection with L. pneumophila was microscopically characterized by the presence of few bacteria inside protozoa at 4th h, and by the beginning of disruptive effects in late phase of trial. In water microcosm studies, performed at different temperature, the more significant interactions were observed at 30°C. In these conditions, L. pneumophila caused a marked reduction in trophozoite and cyst counts from the 4th day until the end of incubation (11 days). B. cepacia showed, by microscopic observation, few and generally single rods within protozoan phagosomes and caused a light reduction of trophozoite viability and cyst formation in co-cultures. A more invasive type of endocytosis, characterized by an early invasion with the presence of a high bacteria number inside amoebae, was observed for Pseudomonas strains. P. fluorescens produced a violent lysis of the host, whereas P. aeruginosa did not cause lysis or suffering. These results underline that water bacteria other than legionella are capable of intracellular survival in Acanthamoeba, influencing the protozoa viable cycle.

  20. Acanthamoeba castellanii: in vitro effects of selected biological, physical and chemical factors.

    PubMed

    Chomicz, Lidia; Padzik, Marcin; Graczyk, Zofi; Starosciak, Bohdan; Graczyk, Thaddeus K; Naprawska, Agnieszka; Oledzka, Gabriela; Szostakowska, Beata

    2010-09-01

    Trophozoites and cysts of free-living Acanthamoeba castellanii present a serious risk to human health as causative agents of human diseases such as fatal granulomatous amoebic encephalitis and Acanthamoeba keratitis that is reported from various part of the world, also in Poland, with increasing frequency, particularly in the contact lens wearers. The amphizoic amoebae are generally extremely resistant to different chemical agents, however, several strains/isolates within A. castellanii may differ in virulence. Among the features considered as associated with the amoeba pathogenicity, temperature tolerance and resistance to different environmental conditions are reported. In the present study, A. castellanii strain cultured in 26 degrees C after several year passages were tested for sensibility/tolerance to instant temperature changes as well as exposition to deuterium oxide, D2O. Significant decrease of number of viable amoebae during in vitro exposition to D2O occurred, but no changes in trophozoites/cysts ratio. The ability of the strain examined to develop in higher temperature may indicate a wide adaptation reserve and its pathogenic potential.

  1. Morphological Features and In Vitro Cytopathic Effect of Acanthamoeba griffini Trophozoites Isolated from a Clinical Case

    PubMed Central

    González-Robles, Arturo; Salazar-Villatoro, Lizbeth; Omaña-Molina, Maritza; Reyes-Batlle, Maria; Martín-Navarro, Carmen M.; Lorenzo-Morales, Jacob

    2014-01-01

    Light and transmission electron microscopy observations are reported on the structure and in vitro cytopathic effect of Acanthamoeba griffini trophozoites isolated from a clinical case. Live trophozoites were moderately active with a remarkable pleomorphism which changed from ovoid to quite elongated shapes. When moving, amoebae formed cytoplasmic projections such as wide lamellae and acanthopodia of diverse size and thickness which contain a significant amount of actin. Ultrastructurally, the cytoplasm showed the main organelles found in other free-living amoebae. Coincubation of trophozoites with MDCK cell monolayers resulted in a local damage to target cells after 24 h of interaction, suggesting that the cytopathic effect is contact-dependent. By transmission electron microscopy, amoebae appeared to engulf small portions of the MDCK cells; however, the cells that were not in contact with trophozoites had an unaltered morphology. When epithelial monolayers were incubated with conditioned medium for 24 h, small areas of cell injury were also observed. The phylogenetical analysis as well as the sequencing of the acquired amplified product for the DF3 region of the amoebae isolate confirmed that it belongs to genotype T3, which includes other pathogenic amoebae; besides the activity of two drugs currently used against Acanthamoeba was tested on A. griffini. PMID:25313337

  2. Recovery of an environmental Chlamydia strain from activated sludge by co-cultivation with Acanthamoeba sp.

    PubMed

    Collingro, Astrid; Poppert, Sven; Heinz, Eva; Schmitz-Esser, Stephan; Essig, Andreas; Schweikert, Michael; Wagner, Michael; Horn, Matthias

    2005-01-01

    Chlamydiae are a unique group of obligate intracellular bacteria comprising important pathogens of vertebrates as well as symbionts of free-living amoebae. Although there is ample molecular evidence for a huge diversity and wide distribution of chlamydiae in nature, environmental chlamydiae are currently represented by only few isolates. This paper reports the recovery of a novel environmental chlamydia strain from activated sludge by co-cultivation with Acanthamoeba sp. The recovered environmental chlamydia strain UV-7 showed the characteristic morphology of chlamydial developmental stages as revealed by electron microscopy and was identified as a new member of the family Parachlamydiaceae (98.7 % 16S rRNA sequence similarity to Parachlamydia acanthamoebae). Infection studies suggested that Parachlamydia sp. UV-7 is not confined to amoeba hosts but is also able to invade mammalian cells. These findings outline a new straightforward approach to retrieving environmental chlamydiae from nature without prior, tedious isolation and cultivation of their natural host cells, and lend further support to suggested implications of environmental chlamydiae for public health.

  3. Influence of Acanthamoeba castellanii on intracellular growth of different Legionella species in human monocytes.

    PubMed

    Neumeister, B; Reiff, G; Faigle, M; Dietz, K; Northoff, H; Lang, F

    2000-03-01

    Previous studies using a murine model of coinhalation of Legionella pneumophila and Hartmannella vermiformis have shown a significantly enhanced intrapulmonary growth of L. pneumophila in comparison to inhalation of legionellae alone (J. Brieland, M. McClain, L. Heath, C. Chrisp, G. Huffnagle, M. LeGendre, M. Hurley, J. Fantone, and C. Engleberg, Infect. Immun. 64:2449-2456, 1996). In this study, we introduce an in vitro coculture model of legionellae, Mono Mac 6 cells (MM6) and Acanthamoeba castellanii, using a cell culture chamber system which separates both cell types by a microporous polycarbonate membrane impervious to bacteria, amoebae, and human cells. Whereas L. pneumophila has shown a maximal 4-log-unit multiplication within MM6, which could not be further increased by coculture with Acanthamoeba castellanii, significantly enhanced replication of L. gormanii, L. micdadei, L. steigerwaltii, L. longbeachae, and L. dumoffii was seen after coculture with amoebae. This effect was seen only with uninfected amoebae, not with Legionella-infected amoebae. The supporting effect for intracellular multiplication in MM6 could be reproduced in part by addition of a cell-free coculture supernatant obtained from a coincubation experiment with uninfected A. castellanii and Legionella-infected MM6, suggesting that amoeba-derived effector molecules are involved in this phenomenon. This coculture model allows investigations of molecular and biochemical mechanisms which are responsible for the enhancement of intracellular multiplication of legionellae in monocytic cells after interaction with amoebae.

  4. Identification of Protein Arginine Methyltransferase 5 as a Regulator for Encystation of Acanthamoeba

    PubMed Central

    Moon, Eun-Kyung; Hong, Yeonchul; Chung, Dong-Il; Goo, Youn-Kyoung; Kong, Hyun-Hee

    2016-01-01

    Encystation is an essential process for Acanthamoeba survival under nutrient-limiting conditions and exposure to drugs. The expression of several genes has been observed to increase or decrease during encystation. Epigenetic processes involved in regulation of gene expression have been shown to play a role in several pathogenic parasites. In the present study, we identified the protein arginine methyltransferase 5 (PRMT5), a known epigenetic regulator, in Acanthamoeba castellanii. PRMT5 of A. castellanii (AcPRMT5) contained domains found in S-adenosylmethionine-dependent methyltransferases and in PRMT5 arginine-N-methyltransferase. Expression levels of AcPRMT5 were increased during encystation of A. castellanii. The EGFP-PRMT5 fusion protein was mainly localized in the nucleus of trophozoites. A. castellanii transfected with siRNA designed against AcPRMT5 failed to form mature cysts. The findings of this study lead to a better understanding of epigenetic mechanisms behind the regulation of encystation in cyst-forming pathogenic protozoa. PMID:27180570

  5. Activity assessment of Tunisian olive leaf extracts against the trophozoite stage of Acanthamoeba.

    PubMed

    Sifaoui, Ines; López-Arencibia, Atteneri; Martín-Navarro, Carmen Ma; Chammem, Nadia; Mejri, Mondher; Lorenzo-Morales, Jacob; Abderabba, Manef; Piñero, José E

    2013-08-01

    The olive tree (Olea europaea, Oleaceae) has historically provided huge economic and nutritional benefits to the Mediterranean basin. In fact, olive leaf extracts have also been used by native people of this area in folk medicine to treat fever and other diseases such as malaria. Recently, several studies have focused on the extraction of high-added-value compounds from olive leaves. However, no previous studies have been developed in order to evaluate the activity of these extracts against Acanthamoeba. In the present work, olive leaf extracts from five different Tunisian varieties of olive trees (Chemlali Tataouine, Zarrazi, Toffehi, Dhokkar, and Limouni) were obtained by using three different solvents, and their activity against the trophozoite stage of Acanthamoeba castellanii Neff was screened. The IC50/96 h (50% parasite growth inhibition) was chosen as the appropriate and comparable data to give as previously described. It could be observed that the amoebicidal activity was dose dependent. Trophozoite growth was inhibited by all the tested extracts with IC50 ranging from 8.234 ± 1.703 μg/ml for the alcoholic mixture of the Dhokkar extract to 33.661 ± 1.398 μg/ml for the methanolic extract of the Toffehi variety. The activity in fact was affected especially by the tested variety and not by the solvent extraction, the Dhokkar variety being the most active one as mentioned above.

  6. Mechanism of cyst specific protein 21 mRNA induction during Acanthamoeba differentiation.

    PubMed

    Chen, Li; Orfeo, Tom; Gilmartin, Greg; Bateman, Erik

    2004-04-01

    The Acanthamoeba cyst specific protein 21 (CSP21) gene is tightly repressed in growing cells and highly induced early during differentiation into a dormant cyst. This increase is mediated by the rate of transcription of the CSP21 gene as determined by nuclear run-on assays. The promoter region of the CSP21 gene was analyzed by transcript start site mapping and in vitro transcription of wild-type or mutant templates, using extracts from growing cells. A sequence located 3' to a modified TATA box completely inhibits transcription and removal of this region permits robust transcription utilizing a start site approximately 35 base pairs downstream of the TATA box. Sequences 5' to the TATA box had no effect on transcription, suggesting that anti-repression is the only mechanism required for CSP21 induction. Fractionation of nuclear extracts yielded a fraction capable of transcription from the CSP21 promoter, and a fraction containing a promoter-specific repressing activity. Anti-repression may thus be a major mechanism regulating differentiation or maintenance of the proliferative cycle in Acanthamoeba.

  7. A riboprinting scheme for identification of unknown Acanthamoeba isolates at species level

    PubMed Central

    Kong, Hyun-Hee

    2002-01-01

    We describe a riboprinting scheme for identification of unknown Acanthamoeba isolates at the species level. It involved the use of PCR-RFLP of small subunit ribosomal RNA gene (riboprint) of 24 reference strains by 4 kinds of restriction enzymes. Seven strains in morphological group I and III were identified at species level with their unique sizes of PCR product and riboprint type by Rsa I. Unique RFCP of 17 strains in group II by Dde I, Taq I and Hae III were classified into: (1) four taxa that were identifiable at the species level, (2) a subgroup of 4 taxa and a pair of 2 taxa that were identical with each other, and (3) a species complex of 7 taxa assigned to A. castellanii complex that were closely related. These results were consistent with those obtained by 18s rDNA sequence analysis. This approach provides an alternative to the rDNA sequencing for rapid identification of a new clinical isolate or a large number of environmental isolates of Acanthamoeba. PMID:11949210

  8. Acanthamoeba keratitis update-incidence, molecular epidemiology and new drugs for treatment.

    PubMed

    Seal, D V

    2003-11-01

    A reliable figure for the expected incidence of Acanthamoeba keratitis of one per 30000 contact lens wearers per year has now been obtained from a combination of three cohort and three Questionnaire Reporting Surveys; 88% of cases wore hydrogel lenses and 12% wore rigid lenses. This figure now provides a basis for the expected number of cases against which to judge either epidemic outbreaks or effects of prevention with disinfecting solutions, better hygiene, or the use of disposable lenses. Molecular biology of Acanthamoeba has advanced considerably in the last 10 years with new automated sequencing technology. This has allowed the construction of a genotype identification scheme with 13 different genotypes against which to compare clinical isolates for epidemiological investigations or pathogenicity markers. So far, only four genotypes have been associated with keratitis of which the majority have been T4 but T3, T6, and T11 have each caused individual cases. Each genotype is heterogenous and can be further subdivided by comparison of sequences of diagnostic fragments of 18S rDNA, riboprinting by PCR-RFLP of 18S rDNA, or by mitochondrial DNA RFLP. Drug therapy has been revolutionised with the introduction of the biguanides-chlorhexidine or polyhexamethylene biguanide-with most but not all infections quickly resolving. Failure can still occur occasionally and further research is needed on more effective combination chemotherapy. A number of guanidines have been identified in this paper that could be usefully pursued as part of combination chemotherapy along with the alkylphosphocholines.

  9. Campylobacter jejuni actively invades the amoeba Acanthamoeba polyphaga and survives within non digestive vacuoles.

    PubMed

    Olofsson, Jenny; Axelsson-Olsson, Diana; Brudin, Lars; Olsen, Björn; Ellström, Patrik

    2013-01-01

    The Gram-negative bacterium Campylobacter jejuni is able to enter, survive and multiply within the free living amoeba Acanthamoeba polyphaga, but the molecular mechanisms behind these events are still unclear. We have studied the uptake and intracellular trafficking of viable and heat killed bacterial cells of the C. jejuni strain 81-176 in A. polyphaga. We found that viable bacteria associated with a substantially higher proportion of Acanthamoeba trophozoites than heat killed bacteria. Furthermore, the kinetics of internalization, the total number of internalized bacteria as well as the intracellular localization of internalized C. jejuni were dramatically influenced by bacterial viability. Viable bacteria were internalized at a high rate already after 1 h of co-incubation and were observed in small vacuoles tightly surrounding the bacteria. In contrast, internalization of heat killed C. jejuni was low at early time points and did not peak until 96 h. These cells were gathered in large spacious vacuoles that were part of the degradative pathway as determined by the uptake of fluorescently labeled dextran. The amount of heat killed bacteria internalized by A. polyphaga did never reach the maximal amount of internalized viable bacteria. These results suggest that the uptake and intracellular survival of C. jejuni in A. polyphaga is bacterially induced.

  10. Investigations of an extraordinary endocytobiont in Acanthamoeba sp.: development and replication.

    PubMed

    Scheid, Patrick; Hauröder, Bärbel; Michel, Rolf

    2010-05-01

    In this article, the results of investigations concerning a parasitic endocytobiont within the host amoebae (Acanthamoeba sp.) are presented. The endocytobiont was recently isolated from the contact lens and the inflamed eye of a patient with keratitis. Light microscopy and electron microscopy were performed to provide morphological details: Light microscopy revealed the presence of ovoid microorganisms developing and proliferating within the cytoplasm of the amoebic trophozoites. Details of the unusual development of these endocytobionts within the amoebae could be studied and demonstrated by means of electron microscopy. Foldings and morphological reorganization of the microorganisms took place exclusively within the host cytoplasm. The intracellularly aggregating organisms led to the rupture of the Acanthamoeba trophozoites after proliferation. Numerous microorganisms were released, which were infectious and were subsequently ingested by hitherto uninfected acanthamoebic trophozoites. To evaluate the in vitro growth of the isolated endocytobionts (without their hosts), they were transferred to several different culture plates. There was no growth of these unique organisms on five different common cultural plates suitable for the growth of bacteria and fungi.

  11. Glycogen phosphorylase in Acanthamoeba spp.: determining the role of the enzyme during the encystment process using RNA interference.

    PubMed

    Lorenzo-Morales, Jacob; Kliescikova, Jarmila; Martinez-Carretero, Enrique; De Pablos, Luis Miguel; Profotova, Bronislava; Nohynkova, Eva; Osuna, Antonio; Valladares, Basilio

    2008-03-01

    Acanthamoeba infections are difficult to treat due to often late diagnosis and the lack of effective and specific therapeutic agents. The most important reason for unsuccessful therapy seems to be the existence of a double-wall cyst stage that is highly resistant to the available treatments, causing reinfections. The major components of the Acanthamoeba cyst wall are acid-resistant proteins and cellulose. The latter has been reported to be the major component of the inner cyst wall. It has been demonstrated previously that glycogen is the main source of free glucose for the synthesis of cellulose in Acanthamoeba, partly as glycogen levels fall during the encystment process. In other lower eukaryotes (e.g., Dictyostelium discoideum), glycogen phosphorylase has been reported to be the main tool used for glycogen breakdown in order to maintain the free glucose levels during the encystment process. Therefore, it was hypothesized that the regulation of the key processes involved in the Acanthamoeba encystment may be similar to the previously reported regulation mechanisms in other lower eukaryotes. The catalytic domain of the glycogen phosphorylase was silenced using RNA interference methods, and the effect of this phenomenon was assessed by light and electron microscopy analyses, calcofluor staining, expression zymogram assays, and Northern and Western blot analyses of both small interfering RNA-treated and control cells. The present report establishes the role of glycogen phosphorylase during the encystment process of Acanthamoeba. Moreover, the obtained results demonstrate that the enzyme is required for cyst wall assembly, mainly for the formation of the cell wall inner layer.

  12. Occurrence and molecular characterization of free-living amoeba species (Acanthamoeba, Hartmannella, and Saccamoeba limax) in various surface water resources of Iran.

    PubMed

    Mahmoudi, Mohammad Reza; Rahmati, Behnaz; Seyedpour, Seyed Hosssen; Karanis, Panagiotis

    2015-12-01

    This study was conducted to determine the presence and molecular identity of Acanthamoeba species in the surface water resources of four provinces in Iran, namely Guilan, Mazandaran (North of Iran), Alborz, and Tehran (capital city), using culture- and molecular-based methods. During March to November 2014, 49 surface water samples were collected from environmental water sources-the distinct surface waters of Guilan, Mazandaran, Alborz, and Tehran provinces, in Iran. For the isolation of Acanthamoeba species, approximately 500 ml of the water samples were filtered through a cellulose nitrate membrane with a pore size of 0.45 μ. The filter was transferred onto non-nutrient agar plates seeded with Gram-negative bacteria (Escherichia coli) as a food source. The presence of Acanthamoeba was confirmed by the genus-specific primer pair JDP1 and 2, and/or NA primers were used to identify Acanthamoeba and certain other free-living amoebae. In total, 38 out of 49 samples were positive by culture and/or PCR for Acanthamoeba and other free-living amoebae from all three provinces. By sequencing the positive isolates, the strains were shown to belong to Acanthamoeba (16 isolates belonged to T4 and 2 isolates belonged to T5), Hartmannella vermiformis (3/24), and Saccamoeba limax (2/24). The T4 and T5 genotypes were detected in Guilan and Mazandaran provinces. Two isolates from Guilan and Tehran provinces belonged to S. limax, and H. vermiformis was detected in Guilan province. The results of this study highlight the need to pay more attention to free-living amoebae, as human activity was observed in all of the localities from which these samples were taken. These surface waters can be potential sources for the distribution and transmission of pathogenic Acanthamoeba in the study areas, and free-living amoebas (FLA) (particularly the Acanthamoeba species) can serve as hosts for and vehicles of various microorganisms.

  13. Genotyping of Acanthamoeba spp. and characterization of the prevalent T4 type along with T10 and unassigned genotypes from amoebic keratitis (AK) patients in India.

    PubMed

    Behera, Himansu Sekhar; Panda, Anita; Satpathy, Gita; Bandivadekar, Pooja; Vanathi, Murgesan; Agarwal, Tushar; Nayak, Niranjan; Tandon, Radhika

    2016-02-16

    Free living amoeba of genus "Acanthamoeba" are the causative agents of severe sight threatening infection of cornea. This study was designed to characterise the genotype of 20 Acanthamoeba spp. isolates obtained from corneal scrapings of 183 suspected Acanthamoeba keratitis (AK) patients reporting to the outpatient department/ causuality services of Dr RP Centre for Ophthalmic Sciences, AIIMS, New Delhi, India in the past 5 years. Corneal scrapings were inoculated onto 2% non-nutrient agar plates overlaid with Escherichia coli. and incubated at 30°C for 15 days. Among 183 suspected patients of Acanthamoeba keratitis 29 were found culture positive for Acanthamoeba spp. out of which 20 samples were established in axenic culture for molecular analysis. DNA was isolated and PCR assay was performed for the amplification of Diagnostic fragment 3(DF3) (~280bp) region of 18S rRNA gene from axenic culture of 20 Acanthamoeba spp. isolates. Rns genotyping was performed on the basis of variation in nucleotide sequences of DF3 region of 18S rRNA gene. In phylogenetic analysis, 16 of the 20 isolates were found to be of prevalent genotype T4, 2 were of genotype T10 and rest 2 of the isolates were of unassigned genotypes. Hence it was concluded that, genotype T4 was found as the most predominant genotype involved in Acanthamoeba keratitis infections. Genotype T10 which was not yet reported from India was detected for the first time in 2 patients. Two isolates were found to be unique, which shared <95% homology with all the known genotypes (T1 - T20) of Acanthamoeba spp.

  14. Use of 18S rRNA gene-based PCR assay for diagnosis of acanthamoeba keratitis in non-contact lens wearers in India.

    PubMed

    Pasricha, Gunisha; Sharma, Savitri; Garg, Prashant; Aggarwal, Ramesh K

    2003-07-01

    Identification of Acanthamoeba cysts and trophozoites in ocular tissues requires considerable expertise and is often time-consuming. An 18S rRNA gene-based PCR test, highly specific for the genus Acanthamoeba, has recently been reported in the molecular diagnosis of Acanthamoeba keratitis. This PCR assay was compared with conventional microbiological tests for the diagnosis of Acanthamoeba keratitis. In a pilot study, the PCR conditions with modifications were first tested on corneal scrapings from patients with culture-proven non-contact lens-related Acanthamoeba, bacterial, and fungal keratitis. This was followed by testing of corneal scrapings from 53 consecutive cases of microbial keratitis to determine sensitivity, specificity, and predictive values of the assay. All corneal scrapings from patients with proven Acanthamoeba keratitis showed a 463-bp amplicon, while no amplicon was obtained from patients with bacterial or fungal keratitis. Some of these amplified products were sequenced and compared with EMBL database reference sequences to validate these to be of Acanthamoeba origin. Out of 53 consecutive cases of microbial keratitis included for evaluating the PCR, 10 (18.9%) cases were diagnosed as Acanthamoeba keratitis on the basis of combined results of culture, smear, and PCR of corneal scrapings. Based on culture results as the "gold standard," the sensitivity of PCR was the same as that of the smear (87.5%); however, the specificity and the positive and negative predictive values of PCR were marginally higher than the smear examination (97.8 versus 95.6%, 87.5 versus 77.8%, and 97.8 versus 97.7%) although the difference was not significant. This study confirms the efficacy of the PCR assay and is the first study to evaluate a PCR-based assay against conventional methods of diagnosis in a clinical setting.

  15. Use of 18S rRNA Gene-Based PCR Assay for Diagnosis of Acanthamoeba Keratitis in Non-Contact Lens Wearers in India

    PubMed Central

    Pasricha, Gunisha; Sharma, Savitri; Garg, Prashant; Aggarwal, Ramesh K.

    2003-01-01

    Identification of Acanthamoeba cysts and trophozoites in ocular tissues requires considerable expertise and is often time-consuming. An 18S rRNA gene-based PCR test, highly specific for the genus Acanthamoeba, has recently been reported in the molecular diagnosis of Acanthamoeba keratitis. This PCR assay was compared with conventional microbiological tests for the diagnosis of Acanthamoeba keratitis. In a pilot study, the PCR conditions with modifications were first tested on corneal scrapings from patients with culture-proven non-contact lens-related Acanthamoeba, bacterial, and fungal keratitis. This was followed by testing of corneal scrapings from 53 consecutive cases of microbial keratitis to determine sensitivity, specificity, and predictive values of the assay. All corneal scrapings from patients with proven Acanthamoeba keratitis showed a 463-bp amplicon, while no amplicon was obtained from patients with bacterial or fungal keratitis. Some of these amplified products were sequenced and compared with EMBL database reference sequences to validate these to be of Acanthamoeba origin. Out of 53 consecutive cases of microbial keratitis included for evaluating the PCR, 10 (18.9%) cases were diagnosed as Acanthamoeba keratitis on the basis of combined results of culture, smear, and PCR of corneal scrapings. Based on culture results as the “gold standard,” the sensitivity of PCR was the same as that of the smear (87.5%); however, the specificity and the positive and negative predictive values of PCR were marginally higher than the smear examination (97.8 versus 95.6%, 87.5 versus 77.8%, and 97.8 versus 97.7%) although the difference was not significant. This study confirms the efficacy of the PCR assay and is the first study to evaluate a PCR-based assay against conventional methods of diagnosis in a clinical setting. PMID:12843065

  16. Ultra-high-speed 3D astigmatic particle tracking velocimetry: application to particle-laden supersonic impinging jets

    NASA Astrophysics Data System (ADS)

    Buchmann, N. A.; Cierpka, C.; Kähler, C. J.; Soria, J.

    2014-11-01

    The paper demonstrates ultra-high-speed three-component, three-dimensional (3C3D) velocity measurements of micron-sized particles suspended in a supersonic impinging jet flow. Understanding the dynamics of individual particles in such flows is important for the design of particle impactors for drug delivery or cold gas dynamic spray processing. The underexpanded jet flow is produced via a converging nozzle, and micron-sized particles ( d p = 110 μm) are introduced into the gas flow. The supersonic jet impinges onto a flat surface, and the particle impact velocity and particle impact angle are studied for a range of flow conditions and impingement distances. The imaging system consists of an ultra-high-speed digital camera (Shimadzu HPV-1) capable of recording rates of up to 1 Mfps. Astigmatism particle tracking velocimetry (APTV) is used to measure the 3D particle position (Cierpka et al., Meas Sci Technol 21(045401):13, 2010) by coding the particle depth location in the 2D images by adding a cylindrical lens to the high-speed imaging system. Based on the reconstructed 3D particle positions, the particle trajectories are obtained via a higher-order tracking scheme that takes advantage of the high temporal resolution to increase robustness and accuracy of the measurement. It is shown that the particle velocity and impingement angle are affected by the gas flow in a manner depending on the nozzle pressure ratio and stand-off distance where higher pressure ratios and stand-off distances lead to higher impact velocities and larger impact angles.

  17. Isolation and molecular characterization of potentially pathogenic Acanthamoeba genotypes from diverse water resources including household drinking water from Khyber Pakhtunkhwa, Pakistan.

    PubMed

    Tanveer, Tania; Hameed, Abdul; Muazzam, Ambreen Gul; Jung, Suk-Yul; Gul, Asma; Matin, Abdul

    2013-08-01

    Acanthamoeba, an opportunistic protozoan pathogen, is ubiquitous in nature, and therefore plays a predatory role and helps control microbial communities in the ecosystem. These Acanthamoeba species are recognized as opportunistic human pathogens that may cause blinding keratitis and rare but fatal granulomatous encephalitis. To date, there is not a single report demonstrating Acanthamoeba isolation and identification from environmental sources in Pakistan, and that is the aim of this study. Acanthamoeba were identified by morphological characteristics of their cysts on non-nutrient agar plates seeded with Escherichia coli. Additionally, the polymerase chain reaction (PCR) was performed with genus-specific primers followed by direct sequencing of the PCR product for molecular identification. Furthermore, our PCR and sequencing results confirmed seven different pathogenic and nonpathogenic genotypes, including T2-T10, T4, T5, T7, T15, T16, and T17. To the best of our knowledge, we have identified and isolated Acanthamoeba sp., for the first time, from water resources of Khyber Pakhtunkhwa, Pakistan. There is an urgent need to address (1) the pathogenic potential of the identified genotypes and (2) explore other environmental sources from the country to examine the water quality and the current status of Acanthamoeba species in Pakistan, which may be a potential threat for public health across the country.

  18. Molecular and Morphometric Characterization of Acanthamoeba spp. from Different Water Sources of Northwest Iran as a Neglected Focus, Co-Bordered With the Country of Iraq

    PubMed Central

    Khezri, Aram; Fallah, Esmaeel; Mostafazadeh, Mostafa; Spotin, Adel; Shahbazi, Abbas; Mahami-Oskouei, Mahmoud; Hazratian, Taimuor

    2016-01-01

    Background Acanthamoeba spp. is a free-living opportunistic protozoan parasites, which can be found in tap, fresh and bottled mineral waters, contact lens solutions, soil etc. Objectives The present study is aimed to determine the Acanthamoeba spp. on the basis of their morpho-molecular aspects in different water sources of the West Azerbaijan province, Northwest of Iran. Methods In this cross-sectional study, 60 water samples were collected from rivers and tap waters during June to September 2015. The water samples were filtered through a cellulose nitrate filter and cultured on non-nutrient agar medium. The extracted DNAs were amplified and some ampliqons were sequenced using partial 18S rRNA for genotyping and phylogenetic analyses. Results Twenty-seven (45%) out of 60 water samples were positive to Acanthamoeba spp. using both culture and morphological examinations. In addition, 24 (40%) out of 27 positive samples in culture method were confirmed by PCR to be Acanthamoeba spp. Conclusions A relatively high prevalence of Acanthamoeba spp. in rivers reflects a risk alert for threatening human health in the region. However, well hygienic status of the tap waters considering Acanthamoeba spp. cannot be ignored in western co-border regions of Iran-Iraq. This study can also serve as a platform for further explorations of water sources in Iran and neighboring countries. PMID:28138374

  19. Genetic analyses of Acanthamoeba isolates from contact lens storage cases of students in Seoul, Korea

    PubMed Central

    Yu, Hak-Sun; Choi, Kyung-Hee; Kim, Hyo-Kyung; Kong, Hyun-Hee

    2001-01-01

    We conducted both the small subunit ribosomal DNA (SSU rDNA) polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) and mitochondrial (mt) DNA RFLP analyses for a genetic characterization of Acanthamoeba isolates from contact lens storage cases of students in Seoul, Korea. Twenty-three strains of Acanthamoeba from the American Type Culture Collection and twelve clinical isolates from Korean patients were used as reference strains. Thirty-nine isolates from contact lens storage cases were classified into seven types (KA/LS1, KA/LS2, KA/LS4, KA/LS5, KA/LS7, KA/LS18, KA/LS31). Four types (KA/LS1, KA/LS2, KA/LS5, KA/LS18) including 33 isolates were regarded as A. castellanii complex by riboprints. KA/LS1 type was the most predominant (51.3%) in the present survey area, followed by KA/LS2 (20.9%), and KA/LS5 (7.7%) types. Amoebae of KA/LS1 type had the same mtDNA RFLP and riboprint patterns as KA/E2 and KA/E12 strains, clinical isolates from Korean keratitis patients. Amoebae of KA/LS2 type had the identical mtDNA RFLP patterns with A. castellanii Ma strain, a corneal isolate from an American patient as amoebae of KA/LS5 type, with KA/E3 and KA/E8 strains from other Korean keratitis patients. Amoebae of KA/LS18 type had identical patterns with JAC/E1, an ocular isolate from a Japanese patient. Three types, which remain unidentified at species level, were not corresponded with any clinical isolate in their mtDNA RFLP and riboprint patterns. Out of 39 isolates analyzed in this study, mtDNA RFLP and riboprint patterns of 33 isolates (84.6%) were identical to already known clinical isolates, and therefore, they may be regarded as potentially keratopathogenic. These results suggest that contact lens wearers in Seoul should pay more attention to hygienic maintenance of contact lens storage cases for the prevention of Acanthamoeba keratitis. PMID:11441503

  20. Genetic analyses of Acanthamoeba isolates from contact lens storage cases of students in Seoul, Korea.

    PubMed

    Yu, H S; Choi, K H; Kim, H K; Kong, H H; Chung, D I

    2001-06-01

    We conducted both the small subunit ribosomal DNA (SSU rDNA) polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) and mitochondrial (mt) DNA RFLP analyses for a genetic characterization of Acanthamoeba isolates from contact lens storage cases of students in Seoul, Korea. Twenty-three strains of Acanthamoeba from the American Type Culture Collection and twelve clinical isolates from Korean patients were used as reference strains. Thirty-nine isolates from contact lens storage cases were classified into seven types (KA/LS1, KA/LS2, KA/LS4, KA/LS5, KA/LS7, KA/LS18, KA/LS31). Four types (KA/LS1, KA/LS2, KA/LS5, KA/LS18) including 33 isolates were regarded as A. castellanii complex by riboprints. KA/LS1 type was the most predominant (51.3%) in the present survey area, followed by KA/LS2 (20.9%), and KA/LS5 (7.7%) types. Amoebae of KA/LS1 type had the same mtDNA RFLP and riboprint patterns as KA/E2 and KA/E12 strains, clinical isolates from Korean keratitis patients. Amoebae of KA/LS2 type had the identical mtDNA RFLP patterns with A. castellanii Ma strain, a corneal isolate from an American patient as amoebae of KA/LS5 type, with KA/E3 and KA/E8 strains from other Korean keratitis patients. Amoebae of KA/LS18 type had identical patterns with JAC/E1, an ocular isolate from a Japanese patient. Three types, which remain unidentified at species level, were not corresponded with any clinical isolate in their mtDNA RFLP and riboprint patterns. Out of 39 isolates analyzed in this study, mtDNA RFLP and riboprint patterns of 33 isolates (84.6%) were identical to already known clinical isolates, and therefore, they may be regarded as potentially keratopathogenic. These results suggest that contact lens wearers in Seoul should pay more attention to hygienic maintenance of contact lens storage cases for the prevention of Acanthamoeba keratitis.

  1. An intracellular replication niche for Vibrio cholerae in the amoeba Acanthamoeba castellanii

    PubMed Central

    Van der Henst, Charles; Scrignari, Tiziana; Maclachlan, Catherine; Blokesch, Melanie

    2016-01-01

    Vibrio cholerae is a human pathogen and the causative agent of cholera. The persistence of this bacterium in aquatic environments is a key epidemiological concern, as cholera is transmitted through contaminated water. Predatory protists, such as amoebae, are major regulators of bacterial populations in such environments. Therefore, we investigated the interaction between V. cholerae and the amoeba Acanthamoeba castellanii at the single-cell level. We observed that V. cholerae can resist intracellular killing. The non-digested bacteria were either released or, alternatively, established a replication niche within the contractile vacuole of A. castellanii. V. cholerae was maintained within this compartment even upon encystment. The pathogen ultimately returned to its aquatic habitat through lysis of A. castellanii, a process that was dependent on the production of extracellular polysaccharide by the pathogen. This study reinforces the concept that V. cholerae is a facultative intracellular bacterium and describes a new host–pathogen interaction. PMID:26394005

  2. Flow cytometric determination of endocytosis of viable labelled Legionella pneumophila by Acanthamoeba palestinensis.

    PubMed

    Harf, C; Goffinet, S; Meunier, O; Monteil, H; Colin, D A

    1997-03-01

    Endocytosis of fluorescently labelled cells of Legionella pneumophila (L. pneumophila) by free-living Acanthamoeba palestinensis (A. palestinensis) has been studied using flow cytometry. L. pneumophila cells were labelled with CM-DiI, a lipophilic fluorescent probe under conditions that did not modify viability. Coculturing the bacteria with amoebae was accompanied by rapid endocytosis; after 5 min, 90% of the amoebae had internalized bacteria. This percentage remained unchanged during further coculture, but the number of bacteria ingested per amoeba increased. Moreover, the number of ingested bacteria was found to be dependent on the size of the amoeba. The validity of the internalization analyzed by flow cytometry was confirmed by observation using epifluorescence and phase contrast microscopy. CM-DiI labelling associated with flow cytometry provides a very valuable technique for the determination of bacteria endocytosis by free-living amoeba.

  3. Growth in Acanthamoeba sp. and antibiotic susceptibility of Legionella micdadei isolated from hot spring water samples.

    PubMed

    Furuhata, Katsunori; Ogihara, Kikumi; Okuno, Rumi; Oonaka, Kenji; Fukuyama, Masafumi

    2009-12-01

    As part of an epidemiological study on legionellosis, we attempted to isolate Legionella spp. from hot spring water samples, and were able to isolate Legionella micdadei from 3 (5.5%) of 55 samples. All of these isolates were able to grow within Acanthamoeba sp., suggesting that the isolates will be pathogens. We also confirmed that the K-2 strain from hot spring water grew in guinea pig monocytes. Sensitivity tests using 10 drugs showed that the isolates were most sensitive to imipenem, with the MIC90 of 0.032 microg/ml, were least sensitive to minocycline, with the MIC90 of 4 microg/ml, and were not sensitive to low amounts of other drugs.

  4. Raman spectroscopic study on the excystation process in a single unicellular organism amoeba (Acanthamoeba polyphaga)

    NASA Astrophysics Data System (ADS)

    Lin, Yu-Chung; Perevedentseva, Elena; Cheng, Chia-Liang

    2015-05-01

    An in vivo Raman spectroscopic study of amoeba (Acanthamoeba polyphaga) is presented. The changes of the spectra during the amoeba cyst activation and excystation are analyzed. The spectra show the changes of the relative intensities of bands corresponding to protein, lipid, and carotenoid components during cyst activation. The presence of carotenoids in the amoeba is observed via characteristic Raman bands. These signals in the Raman spectra are intense in cysts but decrease in intensity with cyst activation and exhibit a correlation with the life cycle of amoeba. This work demonstrates the feasibility of using Raman spectroscopy for the detection of single amoeba microorganisms in vivo and for the analysis of the amoeba life activity. The information obtained may have implications for the estimation of epidemiological situations and for the diagnostics and prognosis of the development of amoebic inflammations.

  5. Raman spectroscopic study on the excystation process in a single unicellular organism amoeba (Acanthamoeba polyphaga).

    PubMed

    Lin, Yu-Chung; Perevedentseva, Elena; Cheng, Chia-Liang

    2015-05-01

    An in vivo Raman spectroscopic study of amoeba (Acanthamoeba polyphaga) is presented. The changes of the spectra during the amoeba cyst activation and excystation are analyzed. The spectra show the changes of the relative intensities of bands corresponding to protein, lipid, and carotenoid components during cyst activation. The presence of carotenoids in the amoeba is observed via characteristic Raman bands. These signals in the Raman spectra are intense in cysts but decrease in intensity with cyst activation and exhibit a correlation with the life cycle of amoeba. This work demonstrates the feasibility of using Raman spectroscopy for the detection of single amoeba microorganisms in vivo and for the analysis of the amoeba life activity. The information obtained may have implications for the estimation of epidemiological situations and for the diagnostics and prognosis of the development of amoebic inflammations.

  6. Acanthamoeba keratitis associated with tap water use during contact lens cleaning: manufacturer guidelines need to change.

    PubMed

    Legarreta, John E; Nau, Amy C; Dhaliwal, Deepinder K

    2013-03-01

    Contact lens-associated Acanthamoeba keratitis continues to be a significant cause of visual morbidity in the United States. Although exposure to water sources while wearing lenses has been a known risk factor for infection for decades, this behavior in several contact lens hygiene protocols continues to prevail. In this review, we surveyed the currently available contact lens cleaning solutions for both soft and rigid gas-permeable contact lenses and reviewed the cleaning instructions of the available solutions. Discrepancies between clinician recommendations and written instructions on a solution packages continues to persist, and we advocate a revision in current manufacturer guidelines to include explicit warnings against use of tap or distilled water sources for cleaning contact lenses or their storage cases.

  7. Amebicidal activity of the essential oils of Lippia spp. (Verbenaceae) against Acanthamoeba polyphaga trophozoites.

    PubMed

    Santos, Israel Gomes de Amorim; Scher, Ricardo; Rott, Marilise Brittes; Menezes, Leociley Rocha; Costa, Emmanoel Vilaça; Cavalcanti, Sócrates Cabral de Holanda; Blank, Arie Fitzgerald; Aguiar, Jaciana dos Santos; da Silva, Teresinha Gonçalves; Dolabella, Silvio Santana

    2016-02-01

    Amoebic keratitis and granulomatous amoebic encephalitis are caused by some strains of free-living amoebae of the genus Acanthamoeba. In the case of keratitis, one of the greatest problems is the disease recurrence due to the resistance of parasites, especially the cystic forms, to the drugs that are currently used. Some essential oils of plants have been used as potential active agents against this protist. Thus, the aim of this study was to determine the amebicidal activity of essential oils from plants of the genus Lippia against Acanthamoeba polyphaga trophozoites. To that end, 8 × 10(4) trophozoites were exposed for 24 h to increasing concentrations of essential oils from Lippia sidoides, Lippia gracilis, Lippia alba, and Lippia pedunculosa and to their major compounds rotundifolone, carvone, and carvacrol. Nearly all concentrations of oils and compounds showed amebicidal activity. The IC50 values for L. sidoides, L. gracilis L. alba, and L. pedunculosa were found to be 18.19, 10.08, 31.79, and 71.47 μg/mL, respectively. Rotundifolone, carvacrol, and carvone were determined as the major compounds showing IC50 of 18.98, 24.74, and 43.62 μg/mL, respectively. With the exception of oil from L. alba, the other oils evaluated showed low cytotoxicity in the NCI-H292 cell line. Given these results, the oils investigated here are promising sources of compounds for the development of complementary therapy against amoebic keratitis and granulomatous amoebic encephalitis and can also be incorporated into cleaning solutions to increase their amebicidal efficiency.

  8. Public health implications of Acanthamoeba and multiple potential opportunistic pathogens in roof-harvested rainwater tanks.

    PubMed

    Hamilton, K A; Ahmed, W; Palmer, A; Sidhu, J P S; Hodgers, L; Toze, S; Haas, C N

    2016-10-01

    A study of six potential opportunistic pathogens (Acanthamoeba spp., Legionella spp., Legionella longbeachae, Pseudomonas aeruginosa, Mycobacterium avium and Mycobacterium intracellulare) and an accidental human pathogen (Legionella pneumophila) in 134 roof-harvested rainwater (RHRW) tank samples was conducted using quantitative PCR (qPCR). All five opportunistic pathogens and accidental pathogen L. pneumophila were detected in rainwater tanks except Legionella longbeachae. Concentrations ranged up to 3.1×10(6) gene copies per L rainwater for Legionella spp., 9.6×10(5) gene copies per L for P. aeruginosa, 6.8×10(5) gene copies per L for M. intracellulare, 6.6×10(5) gene copies per L for Acanthamoeba spp., 1.1×10(5) gene copies per L for M. avium, and 9.8×10(3) gene copies per L for L. pneumophila. Among the organisms tested, Legionella spp. (99% tanks) were the most prevalent followed by M. intracellulare (78%). A survey of tank-owners provided data on rainwater end-uses. Fecal indicator bacteria (FIB) Escherichia coli and Enterococcus spp. were enumerated using culture-based methods, and assessed for correlations with opportunistic pathogens and L. pneumophila tested in this study. Opportunistic pathogens did not correlate well with FIB except E. coli vs. Legionella spp. (tau=0.151, P=0.009) and E. coli vs. M. intracellulare (tau=0.14, P=0.015). However, M. avium weakly correlated with both L. pneumophila (Kendall's tau=0.017, P=0.006) and M. intracellulare (tau=0.088, P=0.027), and Legionella spp. also weakly correlated with M. intracellulare (tau=0.128, P=0.028). The presence of these potential opportunistic pathogens in tank water may present health risks from both the potable and non-potable uses documented from the current survey data.

  9. Insights into the prominent effect of mahanimbine on Acanthamoeba castellanii: Cell profiling analysis based on microscopy techniques

    NASA Astrophysics Data System (ADS)

    Hashim, Fatimah; Amin, Nakisah Mat

    2017-02-01

    Mahanimbine (MH), has been shown to have antiamoeba properties. Therefore, the aim of this study was to assess the growth inhibitory mechanisms of MH on Acanthamoeba castel