The cornea and conjunctiva of the human eye are exposed to external environment and thus are damageable. If the damaged part is infected with some pathogenic microorganisms. serious visual loss may be occured by inflammation. Keratitis or conjunctivitis does not always occur if the eyes are routinely exposed to pathogenic factors because lysozyme in human tears has antimicrobial activity against the microorganisms. 10 this study we have selected 5 strains causing keratitis and/or conjunctivitis. and cultured them in the optimum media. And then we have estimated the growth inhibition of the strains with the addition of various concentration of lysozyme to media to investigate the antimicrobial activity of lysozyme. The results are as follows. The growth of the strains were decreased according to the increase of lysozyme concentration. The growth of Pseudomonas. Neisseria. Klebsiella and Staphylococcus were inhibited 43%, 41%, 35% and 22% respectively by 1 mM concentration of lysozyme. The susceptibility of the gram-negative bacteria to lysozyme is 1.5~2 times higher than the Staphylococcus which is gram-positive bacteria in 1 mM concentration of lysozyme. But lysozyme inhibited the growth of Fusarium which is fungi slightly.
Although the number of patients with Acanthamoeba keratitis has increased dramatically since the widespread use of contact lens, it is still very hard to cure the disease. The proteases from the Acanthamoeba were reported to play important role in the pathogenesis of keratitis. In this study, the inhibitors for extracellular serine proteases of A. castellanii were screened from the extracts of 230 mushroom samples collected from various regions of Korea. The mushrooms were extracted with methanol and water ($65^{\circ}C$). Filtered and concentrated extracts (0.3 mg/ml) were preincubated with proteases before addition of peptide substrate N-succinyl-ala-ala-pro-phe p-anilide. The selected extracts showing strong inhibitory effects were characterized. Although inhibition with single extract was not so high enough, the complete inhibition was achieved with combination of two extracts. The selected extract showed little effect on other serine proteases such as thrombin (human and bovine) and on general protease such as protease K.
Objective: This experimental study was designed to investigate the safety and the effectiveness of Samjeong pharmacopuncture solution (SPS) manufactured by using a the lowtemperature extract on process. Methods: To identify the safety and the effectiveness of using SPS as eye drops, we performed applied eye irritation tests on rabbits and antibacterial tests for Staphylococcus aureus, Staphylococcus epidermidis, Pseudomonas aeruginosa, Aspergillus niger, Fusarium oxysporum, and Candida albicans. The eye irritation test was performed according to the toxicity testing regulation of the Korea Food & Drug Administration (2009. 8. 24, KFDA 2009-116). After SPS had been applied on the left eye of the rabbits, eye irritation in the cornea, iris and conjunctiva was observed on the 1st, 2nd, 3rd, 4th & 7th day. After SPS had been dropped on bacterial species that cause keratitis, the minimum inhibition concentration and the size of the inhibition zone were measured. The anti-bacterial potency was also measured by taking the size of inhibition zone. Results: After SPS had been administered on the left eye of the rabbits, none of nine rabbits were found to show abnormal signs or weight changes. After SPS had been administered on the left eye of the rabbits, no eye irritation in the cornea, iris and conjunctiva was observed on the 1st, 2nd, 3rd, 4th & 7th day. No specific response was detected in MIC for bacterial species Staphylococcus aureus, Staphylococcus epidermidis, Pseudomonas aeruginosa, Aspergillus niger, Fusarium oxysporum, and Candida albicans after SPS had been applied. Conclusions: This study suggests that SPS is a non-toxic and non-irritant medicine that does not cause any of eye irritation in rabbits, but it has no antibacterial effects on bacterial species that are well known to cause keratitis. These results suggest that more research is required on extracts from herbal medicines for treating keratitis.
Kim, Min-Jeong;Quan, Fu-Shi;Kong, Hyun-Hee;Kim, Jong-Hyun;Moon, Eun-Kyung
Parasites, Hosts and Diseases
/
v.60
no.2
/
pp.143-147
/
2022
Acanthamoeba keratitis (AK) is a rare ocular disease, but it is a painful and sight-threatening infectious disease. Early diagnosis and adequate treatment are necessary to prevent serious complications. While AK is frequently diagnosis via several PCR assays or Acanthamoeba-specific antibodies, a more specific and effective diagnostic method is required. This study described the production of a polyclonal peptide antibody against the periplasmic binding protein (PBP) of A. castellanii and investigated its diagnostic potential. Western blot analysis showed that the PBP antibody specifically reacted with the cell lysates of A. castellanii. However, the PBP antibody did not interact with human corneal epithelial (HCE) cells and the other 3 major causative agents of keratitis. Immunocytochemistry (ICC) results revealed the specific detection of A. castellanii trophozoites and cysts by PBP antibodies when A. castellanii were co-cultured with HCE cells. PBP antibody specificity was further confirmed by co-culture of A. castellanii trophozoites with F. solani, S. aureus, and P. aeruginosa via ICC. The PBP antibody specifically reacted with the trophozoites and cysts of A. polyphaga, A. hatchetti, A. culbertsoni, A. royreba, and A. healyi, thus demonstrated its genus-specific nature. These results showed that the PBP polyclonal peptide antibody of A. castellanii could specifically detect several species of Acanthamoeba, contributing to the development of an effective antibody-based AK diagnostics.
The Journal of Korean Medicine Ophthalmology and Otolaryngology and Dermatology
/
v.19
no.1
/
pp.123-134
/
2006
Objective : This experimental study was performed to investigate the effect of Herbe-medicine used for eye disease(Sean-tang, Jinpi-san, Tangpo-san, Copitdis Rhizoma) on Staphylococcus epidermidis keratitis. Methods : After administering various herbal eye drops on Staphylococcus epidermidis, I measured MIC and the size of inhibition zone. MIC was measured by dropping from 20 to $50{\mu}{\ell}$ according to density. Anti-bacterial potency was measured by the size of inhibition zone with change of volume under 2 days culture condition. Also continuous anti-bacterial potency of herbal eye drops was measured by the size of inhibition zone according to 2 days and 6 days culture each under the $50{\mu}{\ell}$ condition. Results : 1. MIC on Staphylococcus epidermidis in Sean-tang, Jinpi-san was 1%, $50{\mu}{\ell}$ 2. MIC on Staphylococcus epidermidis in Tangpo-san, Coptidis rhizoma was 10%, $50{\mu}{\ell}$ 3. MIC on Staphylococcus epidermidis in Cravit was 0.1%, $20{\mu}{\ell}$. 4. Under the 2 days culture condition, the size of inhibition zone on Staphylococcus epidermidis by volume for Sean-tang was 10.0mm in $50{\mu}{\ell}$, Jinpi-san was 16.0mm in $50{\mu}{\ell}$, Tangpo-san was 17.5mm in $50{\mu}{\ell}$, Coptidis rhizoma was 31.0mm in $50{\mu}{\ell}$ and Cravit was 34mm in $50{\mu}{\ell}$, showed the highest anti-bacterial potency. 5. Under the $50{\mu}{\ell}$ condition, the size of inhibition zone on Staphylococcus epidermidis by 2 and 6 culture days for Sean-tang was 47.5mm in 6days, Jinpi-san was 36.0mm in 6days, Tangpo-san was 45.0mm in 6 days and Cravit was 48.0mm in 6 days, which showed the highest anti-bacterial potency. 6. Under the $50{\mu}{\ell}$ culture condition, the size of inhibition zone on Staphylococcus epidermidis by 2 and 6 culture days for Coptidis rhizoma was 31.0mm in 2 days and 6 days, which showed the same anti-bacteria1 potency. Conclusions : The present author think that Herbe-medicine used for eye disease can be used to cure Staphylococcus epidermidis keratitis and if further study is performed, the use of the Herbe-medicine used for eye disease will be valuable and beneficial in the clinical medicines.
Kim, In-Soo;Seo, Jin-Jong;Kim, Kwang-Gon;Ha, Dong-Ryong;Shin, Min-Koo;Kim, Eui-Su;Jeon, Sang-Yun
The Journal of Internal Korean Medicine
/
v.35
no.3
/
pp.288-297
/
2014
Objectives: The goal of this project was to measure the antibacterial effectiveness of Hwangryunhaedok-tang (barberry root decoction for detoxification) and its constituents. All of the active ingredients including the final product were prepared using high-pressure sterilization for use as medicinal eye drops. The varying forms of Hwangryunhaedok-tang (barberry root decoction for detoxification) were used to treat Staphylococcus epidermidis and Enterococcus faecalis which are found in keratitis. Methods: The antibacterial effect was measured by observing the presence or absence of antimicrobial activity when treated with varying concentrations of Hwangryunhaedok-tang extract. The tests were performed using a dosage of $70{\mu}l$ dosages of 100%, 50%, 10% and 1% the extracted solution by the minimum growth inhibitory concentration measurement. Antimicrobial activity was measured by examining the correlation between dosage strength and bacterial activity from $70{\mu}l$ to $10{\mu}l$ at the same concentration. Results: 1. Hwangryunhaedok-tang (barberry root decoction for detoxification), Phellodendri Cortex, and gardenia didn't show any antimicrobial effects against S. aureus, S. epidermidis, or E. faecalis. 2. Barberry root showed antimicrobial effects against S. aureus and S. epidermidis depending on the levels of concentration but didn't show any antimicrobial effects against E. faecalis. 3. Skullcap showed antimicrobial effects against S. aureus and S. epidermidis when a dosage of 100% extract $70{\mu}l$ was used. However, did not show any antimicrobial effects at all against E. faecalis. Conclusions: Hwangryunhaedok-tang (barberry root decoction for detoxification) and its constituents such as barberry root, phellodendri cortex, skullcap, and gardenia, can be used as an alternative to antibiotic medicinal eye drops to treat keratitis. However, further research on effective uses of and efficient extraction methods are needed.
A 6-year-old spayed female Persian cat presented with a 3-month history of recurrent ulcerative keratitis with noticeable opacification and vascularization of the right cornea. The lesion was nonresponsive to topical antibiotics and to nonsteroidal anti-inflammatory drugs. Ophthalmic examination showed signs of ocular discomfort, such as epiphora and blepharospasm, in the right eye. Biomicroscopic examination revealed an irregular, edematous, vascularized mass with pink to white tissue on the entire cornea and mild conjunctivitis. A tentative diagnosis of feline proliferative eosinophilic keratitis (FPEK) was made on the basis of clinical appearance. Cytologic examination of the cornea showed a mixture of numerous eosinophils and mast cells, which confirmed the original diagnosis of FPEK. The cat was treated with a topical antibiotic-corticosteroid combination, cyclosporine ointment, trifluridine eye drops, and oral Llysine. The clinical signs improved remarkably 18 days after the cat was first examined. The short-term use of corticosteroids and long-term use of cyclosporine and an anti-viral agent resolved the lesion without recurrence of the disease for 1 year.
Objectives : This experimental study was performed to investigate the effect of herbal eye drops(Sean-tang, Jinpi-san, Tangpo-san, Copitdis Rhizoma) on Staphylococcus aureus keratitis. Methods : After administering various herbal eye drops on Staphylococcus aureus, I measured MIC and the size of inhibition zone. MIC was measured by dropping from 20 to $50{\mu}{\ell}$ according to density. Anti-bacterial potency was measured by the size of inhibition zone with change of volume under 2 days culture condition. Also continuous antibacterial potency of herbal eye drops was measured by the size of inhibition zone according to 2 days and 6 days culture each under the $50{\mu}{\ell}$condition. Results : 1. MIC on Staphylococcus aureus in Sean-tang, Jinpi-san and Tangpo-san was 100%, $20{\mu}{\ell}$. 2. MIC on Staphylococcus aureus in Coptidis rhizoma was 10%, $30{\mu}{\ell}$. 3. MIC on Staphylococcus aureus in Cravit was 0.1%, $50{\mu}{\ell}$. 4. Under the 2 days culture condition, the size of inhibition zone on Staphylococcus aureus by volume for Sean-tang was 12.0mm in $50{\mu}{\ell}$, Jinpi-san was 19.0mm in $50{\mu}{\ell}$, Tangpo-san was 15.0mm in $50{\mu}{\ell}$, Coptidis rhizoma was 32.7 mm in $50{\mu}{\ell}$ and Cravit was 31mm in $50{\mu}{\ell}$, Coptidis rhizoma showed the highest anti-bacterial potency. 5. Under the $50{\mu}{\ell}$ condition, the size of inhibition zone on Staphylococcus aureus by 2 and 6 culture days for Sean-tang was 12.0mm, Jinpi-san was 19.0mm, Tangpo-san was 15.0mm, Coptidis rhizoma was 32.7mm and Cravit was 31.0 mm, which showed sameness anti-bacterial potency in 2 days and 6 days. Conclusions : The present author think that the herbal eye drops can be used to cure Staphylococcus aureus keratitis and if further study is performed, the use of the herbal eye drops will be valuable and beneficial in the clinical medicines.
In order to observe the cytotoxicity of Acanthamoeba spp., which were isolated from contact lens containers as ethiological agents for the probable amoebic keratitis in Korea, the crystal violet staining method and LDH release assay were carried out. In the crystal violet staining method, among eight contact lens container isolates, isolate 3 (Acanthauloeba KA/LS5) showed 83.6% and 81.8% of cytotoxicity, and isolate 7 (Acanthamoeba KA/LS37) showed 28.2% and 25.1% of cytotoxicity, in 1 mg/ml and 0.5 mg/ml Iysate treatments, respectively. Acanthamoeba cutbertsoni and A. healyi showed 84.0% and 82.8% of cytotoxicity. Similar results were observed in A. costellunii and A. hafchefti which showed 83.6% and 75.5% or cytotoxicity. Acanthamoeba roureba and A. polyphaga showed 9.0% and 1.7% of cytotoxicity. In the LDH release assay, isolate 3 (20.4%) showed higher cytotoxicity than other isolates in 1 mg/ml Iysate treatment. The results provide that at least isolate 3 has the cytotoxic effect against CHO cells and seems to be the pathogenic strain.
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.
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