scholarly journals Evaluation of the HB&L System for the Microbiological Screening of Storage Medium for Organ-Cultured Corneas

2013 ◽  
Vol 2013 ◽  
pp. 1-5 ◽  
Author(s):  
D. Camposampiero ◽  
S. Grandesso ◽  
E. Zanetti ◽  
S. Mazzucato ◽  
M. Solinas ◽  
...  
Keyword(s):  
Contamination Rate ◽  
Storage Medium ◽  
Human Donor ◽  
Predictive Values ◽  
Anaerobic Microorganisms ◽  
L System ◽  
Microbiological Control ◽  
Corneal Storage ◽  
Aerobic And Anaerobic ◽  
Donor Corneas

Aims. To compare HB&L and BACTEC systems for detecting the microorganisms contaminating the corneal storage liquid preserved at 31°C.Methods. Human donor corneas were stored at 4°C followed by preservation at 31°C. Samples of the storage medium were inoculated in BACTEC Peds Plus/F (aerobic microorganisms), BACTEC Plus Anaerobic/F (anaerobic microorganisms), and HB&L bottles. The tests were performed (a) after six days of storage, (b) end of storage, and (c) after 24 hours of preservation in deturgescent liquid sequentially. 10,655 storage and deturgescent media samples were subjected to microbiological control using BACTEC (6-day incubation) and HB&L (24-hour incubation) systems simultaneously. BACTEC positive/negative refers to both/either aerobic and anaerobic positives/negatives, whereas HB&L can only detect the aerobic microbes, and therefore the positives/negatives depend on the presence/absence of aerobic microorganisms.Results. 147 (1.38%) samples were identified positive with at least one of the two methods. 127 samples (134 identified microorganisms) were positive with both HB&L and BACTEC. 14 HB&L+/BACTEC− and 6 BACTEC+/HB&L− were identified. Sensitivity (95.5%), specificity (99.8%), and positive (90.1%) and negative predictive values (99.9%) were high with HB&L considering a 3.5% annual contamination rate.Conclusion. HB&L is a rapid system for detecting microorganisms in corneal storage medium in addition to the existing methods.

scholarly journals Killing efficacy of a new hypothermic corneal storage medium against the micro-organisms frequently found in human donor cornea intended for transplantation

2021 ◽  
Vol 6 (1) ◽  
pp. e000833
Author(s):  
Laura Giurgola ◽  
Claudio Gatto ◽  
Claudia Honisch ◽  
Orietta Rossi ◽  
Eugenio Ragazzi ◽  
...  
Keyword(s):  
Microbial Count ◽  
Human Donor ◽  
Aspergillus Brasiliensis ◽  
Colony Forming Units ◽  
Corneal Storage ◽  
Hypothermic Storage ◽  
Almost All ◽  
Micro Organisms ◽  
Donor Corneas

ObjectiveTo study the in vitro killing efficacy of Kerasave (AL.CHI.MI.A Srl), a medium provided with amphotericin B tablet for hypothermic storage of human donor corneas, against relevant contaminants associated with postkeratoplasty infections.Methods and AnalysisThe antimicrobial activity of Kerasave was determined after 0, 3 and 14 days of incubation at 2°C–8°C, inoculating Kerasave and the control medium with 105–106 colony forming units (CFU) of Candida albicans (CA), Fusarium solani (FS), Aspergillus brasiliensis (AB), Staphylococcus aureus (SA), Enterococcus faecalis (EF), Bacillus subtilis spizizenii (BS), Pseudomonas aeruginosa (PA), Enterobacter cloacae (EC) and Klebsiella pneumoniae (KP). Log10 reductions at different time intervals were determined by assessing the number of viable CFU using the serial dilution plating technique.ResultsAfter 3 days, Kerasave induced the highest log10 decrease in the concentrations of KP, PA, CA and EC (5.37, 4.15, 2.97 and 2.67, respectively; all p<0.001). The log10 decreases of SA and EF were 2.27 and 2.11, respectively (all p<0.001). The lowest log10 decrease was observed in BS, AB and FS concentrations (0.25, 0.30 and 0.67, respectively; p<0.001 for BS and AB and p=0.004 for FS). After 14 days, the microbial count of CA, FS, SA, EF, PA and EC further decreased (p=0.006 for FS; p<0.001 for the others).ConclusionKerasave effectively reduced or kept unchanged the microbial concentration of almost all tested strains after 3 days. Thus, this novel medium represents a valuable tool to control the microbial contamination of human donor corneas during hypothermic storage for up to 14 days before transplantation.

A new storage medium containing amphotericin B versus Optisol-GS for preservation of human donor corneas

2020 ◽  
pp. bjophthalmol-2020-317136
Author(s):  
Raffaela Mistò ◽  
Laura Giurgola ◽  
Francesca Pateri ◽  
Anna Limongelli ◽  
Eugenio Ragazzi ◽  
...  
Keyword(s):  
Endothelial Cell ◽  
Amphotericin B ◽  
Corneal Thickness ◽  
Storage Period ◽  
Blue Staining ◽  
Specular Microscopy ◽  
Storage Medium ◽  
Human Donor ◽  
Corneal Transparency ◽  
Donor Corneas

Background/AimWe compared the quality of human donor corneas stored in a cold storage medium containing 2.5 μg/ml of amphotericin B (Kerasave, AL.CHI.MI.A. S.R.L., Ponte San Nicolò, Italy) and Optisol-GS (Bausch & Lomb Inc., Bridgewater, NJ, USA) for 14 days.MethodsSixteen pairs of human donor corneas were collected in Eusol-C (AL.CHI.MI.A. S.R.L., Ponte San Nicolò, Italy). Next, all tissues underwent the first evaluation that included the assessments of central corneal thickness (CCT), endothelial cell density (ECD) measured using both trypan blue staining and specular microscopy, endothelial cell (EC) mortality and morphology, and corneal transparency within 24 hours from recovery (Day 1). Afterwards, one cornea of each pair was transferred into Kerasave or Optisol-GS. ECD and CCT were also assessed at Day 7, and all the metrics were evaluated again at the end of the storage period (Day 14).ResultsAt all tested time points, no differences were found in the qualitative (corneal transparency, EC morphology) and quantitative metrics (ECD, CCT, EC mortality) between the Kerasave and the Optisol-GS storage groups. At Day 14, the corneas stored in Kerasave and Optisol-GS showed ECD of 2312±98 and 2335±128 cells/mm2 (p=0.886), CCT of 717±17 and 697±19 μm (p=0.454) and central EC mortality of 0.54%±0.40% and 0.14%±0.14% (p=0.719), respectively.ConclusionsThe new amphotericin B−containing medium Kerasave was comparable to Optisol-GS in terms of preservation of corneal characteristics at 2–8°C for 14 days.

scholarly journals Plant Recombinant Human Collagen Type I Hydrogels for Corneal Regeneration

2021 ◽  
Author(s):  
Michel Haagdorens ◽  
Elle Edin ◽  
Per Fagerholm ◽  
Marc Groleau ◽  
Zvi Shtein ◽  
...  
Keyword(s):  
Collagen Type ◽  
Collagen Type I ◽  
Collagen Fibrils ◽  
Type I ◽  
Safe Alternative ◽  
Human Donor ◽  
Human Collagen ◽  
Donor Corneas

Abstract Purpose To determine feasibility of plant-derived recombinant human collagen type I (RHCI) for use in corneal regenerative implants Methods RHCI was crosslinked with 1-ethyl-3-(3-dimethyl aminopropyl) carbodiimide (EDC) and N-hydroxysuccinimide (NHS) to form hydrogels. Application of shear force to liquid crystalline RHCI aligned the collagen fibrils. Both aligned and random hydrogels were evaluated for mechanical and optical properties, as well as in vitro biocompatibility. Further evaluation was performed in vivo by subcutaneous implantation in rats and corneal implantation in Göttingen minipigs. Results Spontaneous crosslinking of randomly aligned RHCI (rRHCI) formed robust, transparent hydrogels that were sufficient for implantation. Aligning the RHCI (aRHCI) resulted in thicker collagen fibrils forming an opaque hydrogel with insufficient transverse mechanical strength for surgical manipulation. rRHCI showed minimal inflammation when implanted subcutaneously in rats. The corneal implants in minipigs showed that rRHCI hydrogels promoted regeneration of corneal epithelium, stroma, and nerves; some myofibroblasts were seen in the regenerated neo-corneas. Conclusion Plant-derived RHCI was used to fabricate a hydrogel that is transparent, mechanically stable, and biocompatible when grafted as corneal implants in minipigs. Plant-derived collagen is determined to be a safe alternative to allografts, animal collagens, or yeast-derived recombinant human collagen for tissue engineering applications. The main advantage is that unlike donor corneas or yeast-produced collagen, the RHCI supply is potentially unlimited due to the high yields of this production method. Lay Summary A severe shortage of human-donor corneas for transplantation has led scientists to develop synthetic alternatives. Here, recombinant human collagen type I made of tobacco plants through genetic engineering was tested for use in making corneal implants. We made strong, transparent hydrogels that were tested by implanting subcutaneously in rats and in the corneas of minipigs. We showed that the plant collagen was biocompatible and was able to stably regenerate the corneas of minipigs comparable to yeast-produced recombinant collagen that we previously tested in clinical trials. The advantage of the plant collagen is that the supply is potentially limitless.

Comparison of Corneal Storage in K-Sol and Chondroitin Sulfate Corneal Storage Medium in Human Corneal Transplantation

Ophthalmology ◽  
1989 ◽  
Vol 96 (5) ◽  
pp. 688-697 ◽  
Author(s):  
Jonathan H. Lass ◽  
William J. Reinhart ◽  
William E. Bruner ◽  
Mary Lou Kachmer ◽  
Mark D. Lomeo ◽  
...  
Keyword(s):  
Chondroitin Sulfate ◽  
Corneal Transplantation ◽  
Storage Medium ◽  
Corneal Storage

Vaginal microbiomes in women with infertility that was surgical treatment on the uterine appendages

2018 ◽  
pp. 98-102
Author(s):  
A.E. Dubchak ◽  
◽  
A.V. Milevsky ◽  
N.N. Obeid ◽  
◽  
...  
Keyword(s):  
Surgical Treatment ◽  
Antimicrobial Agents ◽  
Inflammatory Diseases ◽  
Reproductive Age ◽  
Anaerobic Microorganisms ◽  
Normal Microflora ◽  
Vaginal Tract ◽  
High Degree ◽  
Aerobic And Anaerobic ◽  
Group 1

The objective: of the study was to study the microbial vaginal tract in women with infertility, who had undergone surgical treatment on the uterine appendages. Materials and methods. An examination of vaginal discharge on flora and STIs was conducted in 120 women of reproductive age with infertility and benign ovarian formations, ectopic pregnancy, which was organ-retained surgical intervention on the appendages of the uterus. Of these, 76 (1 group) patients were surgically treated in a planned manner, 44 (in 2 groups) – in urgent cases. Results. Inflammatory diseases of the genital area were more common in women with infertility, who had surgical treatment in an urgent manner than in women of group 1 (p<0.05), especially inflammatory diseases of the cervix – almost twice as likely as in group 1. The vaginal microbial examination of women with infertility who were hospitalized for surgical treatment in an urgent manner indicates a significant imbalance between the parameters of contamination of the genital tract by conditionally pathogenic and normal microflora. This, above all, was manifested by the high frequency of determination in women of the 2nd group of representatives of optional aerobes and anaerobes (p<0.05). The concentration of facultative-aerobic and anaerobic microorganisms was in patients of the 2nd group, mostly of high degree of microbial dissemination, and in women of the 1 group, medium and low. STIs were found mainly in association with anaerobic and aerobic microorganisms in patients who were operated in an urgent manner. Conclusion. It is necessary to correct the vaginal biotope in women with infertility before surgical treatment – to promptly influence pathogenic microorganisms with antimicrobial agents and restore physiological vaginal microbial vagina. Key words: women, infertility, vaginal microbial, appendages of the uterus, surgical treatment.

Depth-dependent Cohesive Tensile Strength in Human Donor Corneas: Implications for Refractive Surgery

2008 ◽  
Vol 24 (1) ◽  
pp. S85-S89 ◽  
Author(s):  
J. Bradley Randleman ◽  
Daniel G Dawson ◽  
Hans E Grossniklaus ◽  
Bernard E McCarey ◽  
Henry F Edelhauser
Keyword(s):  
Tensile Strength ◽  
Refractive Surgery ◽  
Human Donor ◽  
Donor Corneas

scholarly journals Variability in Arsenic Methylation Efficiency across Aerobic and Anaerobic Microorganisms

2020 ◽  
Vol 54 (22) ◽  
pp. 14343-14351
Author(s):  
Karen Viacava ◽  
Karin Lederballe Meibom ◽  
David Ortega ◽  
Shannon Dyer ◽  
Arnaud Gelb ◽  
...  
Keyword(s):  
Anaerobic Microorganisms ◽  
Arsenic Methylation ◽  
Aerobic And Anaerobic

Organic Acids and Anaerobic Microorganisms in the Contents of the Cholesteatoma SAC

1983 ◽  
Vol 92 (1) ◽  
pp. 91-96 ◽  
Author(s):  
Yukiko Iino ◽  
Tomonori Takasaka ◽  
Etsuro Hoshino ◽  
Yutaka Kaneko ◽  
Sachiko Tomioka ◽  
...  
Keyword(s):  
Fatty Acids ◽  
Organic Acids ◽  
Volatile Fatty Acids ◽  
Chromatographic Technique ◽  
Anaerobic Conditions ◽  
Clostridium Species ◽  
Anaerobic Microorganisms ◽  
Aerobic And Anaerobic ◽  
Aerobic And Anaerobic Conditions

Organic acids in the contents of the cholesteatoma sac from 28 cases were studied by gas chromatographic technique. Five volatile fatty acids (acetate, propionate, isobutyrate, butyrate and isovalerate) and lactate were detected in large amounts, which may lower the pH of the cholesteatoma content. These acids were considered to be derived from products of anaerobic microorganisms. Therefore, the contents from 12 cases were cultured anaerobically in a glove box. Obligate microorganisms were identified in 92% of the cases and Peptococcus, Bacteroides, and Clostridium species were frequently isolated. In vitro, such obligate anaerobes produced various organic acids from the cholesteatoma content. Facultatives such as Staphylococcus aureus and Proteus mirabilis produced acetate in the content under aerobic and anaerobic conditions, whereas no organic acid was produced by Pseudomonas aeruginosa. Organic acids in the cholesteatoma content could be fermentative products made by the microorganisms, anaerobes and facultatives, which use the content as a substrate for acid production.

Comparisons of Cornea Cold, a New Corneal Storage Medium, and Optisol-GS

Cornea ◽  
2020 ◽  
Vol Publish Ahead of Print ◽  
Author(s):  
Joanne W. Ho ◽  
Hoon Jung ◽  
Mimi Chau ◽  
James A. Kuchenbecker ◽  
Michael Banitt
Keyword(s):  
Storage Medium ◽  
Corneal Storage
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