scholarly journals Er:YAG laser-assisted filtration surgery: initial results in rabbits

2021 ◽  
Vol 21 (1) ◽  
Author(s):  
Noa Kapelushnik ◽  
Ari Leshno ◽  
Reut Singer ◽  
Ruth Huna-Baron ◽  
Yaniv Barkana ◽  
...  
Keyword(s):  
Er:Yag Laser ◽  
Laser System ◽  
Pulse Length ◽  
Glaucoma Surgery ◽  
Coagulation Necrosis ◽  
Histopathological Analysis ◽  
Subconjunctival Injection ◽  
A Minor ◽  
Ab Interno

Abstract Background Glaucoma is a leading cause of global blindness, especially preventable blindness. The increased prevalence of glaucoma has led to a growing demand for newer, safer, more rapid, and simpler treatments for the reduction of intraocular pressure (IOP). In this study, we evaluated the safety and feasibility of performing filtration glaucoma surgery with an Ab-Interno Er:YAG laser in rabbits. Methods Nine New Zealand White rabbits age 16 weeks were studied. After subconjunctival injection of mitomycin C (MMC), a novel Ab-Interno Er:YAG laser probe was inserted into the anterior chamber (AC) through a clear corneal 1 mm paracentesis and directed at the trabecular meshwork adjacent to the MMC injection area. A pulsed laser beam was applied to create a patent sclerostomy connecting the AC to the subconjuctival space, resulting in a filtering bleb. The laser system used was the Er:YAG laser system - LAS25-FCU, (Pantec Biosolutions AG, Liechtenstein). Parameters used: Wave lengh: 2940 nm, Pulse length: 100–400 μsec,frequency: 250 Hz. Average laser power in accordance to the fiber tip diameter: 0.85 W(range 0.8–0.92 W). Complete ocular exams, including IOP measurements, were performed on 1, 7, 14, and 23 days postoperatively. Three rabbits were sacrificed on days 1, 14, and 23, and histological examinations were performed on all nine eyes. Results All procedures resulted in a functional medium-large superior bleb without significant complications. The bleb was sustained in all rabbits by day 14 and in one of the three rabbits that reached the last follow-up at 23 days. No cases of postoperative hypotony were observed. There was a transient significant reduction in mean IOP on postoperative days 5 and 7 (P = 0.028). Histopathological analysis revealed a patent full-thickness scleral tunnel with only a minor degree of surrounding coagulation necrosis. Conclusions The Ab-Interno laser sclerostomy procedure is potentially safe and effective based on initial experience in an in-vivo rabbit animal model.

136 IN VITRO DEVELOPMENT OF BOVINE EMBRYOS SUBMITTED TO LASER ASSISTED HATCHING ON DAY 6 AND DAY 7 OF EMBRYO CULTURE

2009 ◽  
Vol 21 (1) ◽  
pp. 168
Author(s):  
C. B. Ponchirolli-Schneider ◽  
J. H. Pryor ◽  
C. R. Looney
Keyword(s):  
Embryo Culture ◽  
Uv Light ◽  
Laser System ◽  
Pulse Length ◽  
Assisted Hatching ◽  
In Vitro Production ◽  
Stage Of Development ◽  
Number Of Cells

In vitro production (IVP) of embryos is a valuable tool in bovine assisted reproduction. IVP embryos show lower pregnancy rates when compared to in vivo produced embryos. The inability of the IVP embryo to hatch from the zona pellucida (ZP) after embryo transfer is believed to be one contributing factor. The aim of this study was to compare the ability of IVP embryos to hatch and develop in vitro after being submitted to laser assisted hatching (LAH) at two different time periods of embryo culture: 144 h (Day 6) and 168 h (Day 7). In vitro maturated oocytes from slaughterhouse ovaries (Ovitra Biotechnologies, Amarillo, TX, USA) were fertilized with frozen/thawed semen from two different bulls (Day 0) and cultured in G1.5/G2.5 medium supplemented with 8 mg mL–1 of BSA (Vitrolife, Englewood, CA, USA) at 38.5°C, 5% CO2, 5% O2, 5% N2, in humidified atmosphere. On Day 6 post-fertilization, all viable embryos (n = 251), from three replicates, were washed in holding medium (Vigro Holding Plus, Bioniche, Pullman, WA, USA) and divided into four treatment groups: laser assisted hatching Day 6 and Day 7 (LAHD6 and LAHD7), control Day 6 and Day 7 (CD6 and CD7). The groups LAHD7 and CD7 were immediately placed in G2.5 and returned to the incubator until Day 7. Embryos from groups LAHD6 and CD6 were placed in G2.5 in separate wells of a four well dish and covered with 300 μL of mineral oil. Embryos from LAHD6 group had one third to one half of the external edge of the ZP exposed to a laser beam, using XY Clone® (Hamilton Thorne Biosciences, Inc., Beverly, MA, USA) laser system, with a pulse strength of 90% and a pulse length of 600 μs. The group CD6 was submitted to the same conditions, but did not receive the laser treatment. On Day 7, the experiment was repeated for embryos belonging to groups LAHD7 and CD7. Embryos from all groups were cultured in vitro and evaluated on Day 8 and Day 9 for stage of development. On Day 9, a random sample of embryos from each treatment group (n = 48) was stained with Hoechst 33342 (2.5 μg mL–1) and evaluated under UV light to determine the total number of cells. The number of hatched blastocysts was not different (chi-square, p > 0.05) among the groups on Day 9 of culture (LAHD6 = 49/66, CD6 = 38/61, LAHD7 = 42/59, CD7 = 47/65; 74, 62, 71 and 72%, respectively). However, on Day 8 of culture, LAHD6 showed a higher number (p < 0.05) of hatched blastocysts (40/66, 60%), when compared to group CD6 (26/61, 42%). There was no difference between groups LAHD7 (33/59, 55%) and CD7 (31/65, 47%) on Day 8. Comparison of the total number of cells showed no difference (Student’s t-test, p > 0.05) among the groups (LAHD6 = 154.8 ± 12.2, CD6 = 184.4 ± 19.5, LAHD7 = 170.4 ± 15.8, CD7 = 162.5 ± 14.6), indicating that LAH does not have a detrimental effect on mean cell production throughout embryo development in vitro. These data show that LAH on Day 6 of culture improves in vitro hatching rates on Day 8, while LAH on Day 7 shows no improvement on either Day 8 or 9. Kathy Bradley, Hamilton Thorne Biosciences, Inc.

Preparation and Biochemical Evaluation of Functionalized Multi-Walled Carbon Nanotubes with Punica granatum Extract

2019 ◽  
Vol 15 (1) ◽  
pp. 138-144 ◽  
Author(s):  
Ahmed A. Haroun ◽  
Abdel-Tawab H. Mossa ◽  
Samia M.M. Mohafrash
Keyword(s):  
Liver Enzymes ◽  
Histopathological Analysis ◽  
Pomegranate Peel ◽  
Liver And Kidney ◽  
Multi Walled Carbon Nanotubes ◽  
Pomegranate Peel Extract ◽  
Walled Carbon Nanotubes ◽  
Functionalized Mwcnts ◽  
Peel Extract

Background: Funcionalized multi-walled carbon nanotubes (ox-MWCNTs) were used for the preparation of therapeutic nanoparticles for delivery of some bioactive compounds. Consequently, this work deals with the preparation of grafted MWCNTs with n-vinyl caprolactam in the presence of pomegranate peel extract (P. granatum), titanium dioxide (TiO2) and/or silver nanoparticeles and their toxic effects on male mice using in vivo biological examination (liver and kidney dysfunction biomarkers) and the histopathological analysis. Methods: P. granatum extract was immobilized onto functionalized MWCNTs using simple adsorption technique. Moreover, The prepared materials were analyzed by Fourier transform infrared spectroscopy (FTIR), scanning electron microscope (SEM). In vivo examination using liver and kidney dysfunction biomarkers was investigated. In addition, the histopathological study was carried out. Results: The ox-MWCNTs induced significant elevation in the liver enzymes including AST, ALT and ALP relative to the control group. While, the treatment with P. granatum extract only did not induce any change in the liver and kidney biomarkers. In other words, P. granatum extract loaded onto functionalized MWCNTs showed low effects on liver enzymes and kidney function biomarkers in the treated mice in comparison with ox-MWCNTs and extract separately. Moreover, histopathological analysis revealed that the P. granatum extract functionalized MWCNTs exhibited normal renal tissue with no histopathological alteration. Conclusion: The grafted MWCNTs with n-vinyl caprolactam in the presence of pomegranate peel extract (P. granatum), titanium dioxide (TiO2) and/or silver nanoparticeles were successfully prepared. SEM-micrographs showed complete coating of MWCNTs fiber with the extract. The prepared materials resulted in no toxic effects and the histopathological findings were confirmed by inflammation of the liver and kidney tissues.

scholarly journals Effect of postoperative corticosteroids on surgical outcome and aqueous autotaxin following combined cataract and microhook ab interno trabeculotomy

2021 ◽  
Vol 11 (1) ◽  
Author(s):  
Megumi Honjo ◽  
Reiko Yamagishi ◽  
Nozomi Igarashi ◽  
Chui Yong Ku ◽  
Makoto Kurano ◽  
...  
Keyword(s):  
Surgical Outcome ◽  
Primary Open Angle Glaucoma ◽  
Open Angle Glaucoma ◽  
Case Series ◽  
Iop Elevation ◽  
Immunoenzymatic Assay ◽  
Postoperative Fibrosis ◽  
Ab Interno

AbstractTo evaluate the effect of postoperative corticosteroids on surgical outcome and autotaxin (ATX) levels after microhook ab interno trabeculotomy combined with cataract surgery (μLOT-CS), prospective, consecutive non-randomized case series comparing outcomes of 30 eyes with primary open angle glaucoma was performed. The aqueous ATX, intraocular pressure (IOP) and glaucoma medications were monitored for 3 months postoperatively. An in-vivo mouse μLOT model was generated. In vitro, ATX and fibrotic changes induced by dexamethasone (Dex) treatment following scratch (S) in cultured human trabecular meshwork (hTM) cells were assessed by immunofluorescence, immunoenzymatic assay, and RT-qPCR. Postoperative ATX at 1 week and the number of antiglaucoma medications at 3 months were significantly lower in non-steroid group, and steroid use was the only variable significantly associated with postoperative medications at 3 months in multiregression analyses. In vitro, ATX activity was significantly upregulated in the Dex + S group, and αSMA was significantly upregulated in the Dex and Dex + S groups. Fibronectin and COL1A1 were significantly upregulated in the S group. μLOT-CS decreased IOP and medications in the overall cohort, and non-use of postoperative steroids resulted in a smaller number of postoperative medications. Limiting postoperative steroids in μLOT may minimize IOP elevation and postoperative fibrosis.

scholarly journals Development of Triamcinolone Acetonide-Loaded Microemulsion as a Prospective Ophthalmic Delivery System for Treatment of Uveitis: In Vitro and In Vivo Evaluation

Pharmaceutics ◽  
2021 ◽  
Vol 13 (4) ◽  
pp. 444
Author(s):  
Alaa Mahran ◽  
Sayed Ismail ◽  
Ayat A. Allam
Keyword(s):  
Triamcinolone Acetonide ◽  
Delivery System ◽  
Histopathological Examination ◽  
Rabbit Model ◽  
Subconjunctival Injection ◽  
In Vivo Evaluation ◽  
Ternary Phase Diagrams ◽  
Ophthalmic Delivery

Treatment of uveitis (i.e., inflammation of the uvea) is challenging due to lack of convenient ophthalmic dosage forms. This work is aimed to determine the efficiency of triamcinolone acetonide (TA)-loaded microemulsion as an ophthalmic delivery system for the treatment of uveitis. Water titration method was used to construct different pseudo-ternary phase diagrams. Twelve microemulsion formulations were prepared using oleic acid, Cremophor EL, and propylene glycol. Among all tested formulations, Formulation F3, composed of oil: surfactant-co-surfactant (1:1): water (15:35:50% w/w, respectively), was found to be stable and showed acceptable pH, viscosity, conductivity, droplet size (211 ± 1.4 nm), and zeta potential (−25 ± 1.7 mV) and almost complete in vitro drug release within 24 h. The in vivo performance of the optimized formulation was evaluated in experimentally uveitis-induced rabbit model and compared with a commercial TA suspension (i.e., Kenacort®-A) either topically or by subconjunctival injection. Ocular inflammation was evaluated by clinical examination, white blood cell count, protein content measurement, and histopathological examination. The developed TA-loaded microemulsion showed superior therapeutic efficiency in the treatment of uveitis with high patient compliance compared to commercial suspension. Hence, it could be considered as a potential ocular treatment option in controlling of uveitis.

scholarly journals Comparison of Efficacy between 120° and 180° Schlemm’s Canal Incision Microhook Ab Interno Trabeculotomy

2021 ◽  
Vol 10 (14) ◽  
pp. 3181
Author(s):  
Naoki Okada ◽  
Kazuyuki Hirooka ◽  
Hiromitsu Onoe ◽  
Yumiko Murakami ◽  
Hideaki Okumichi ◽  
...  
Keyword(s):  
Surgical Outcomes ◽  
Open Angle Glaucoma ◽  
Case Series ◽  
Glaucoma Surgery ◽  
Success Rates ◽  
Schlemm’S Canal ◽  
Schlemm's Canal ◽  
Kaplan Meier ◽  
The Mean ◽  
Ab Interno

We compared surgical outcomes in patients with either primary open-angle glaucoma or exfoliation glaucoma after undergoing combined phacoemulsification with either a 120° or 180° incision during a Schlemm’s canal microhook ab interno trabeculotomy (μLOT-Phaco). This retrospective comparative case series examined 52 μLOT-Phaco eyes that underwent surgery between September 2017 and December 2020. Surgical qualified success was defined as an intraocular pressure (IOP) of ≤20 mmHg, ≥20% IOP reduction with IOP-lowering medications, and no additional glaucoma surgery. Success rates were evaluated by Kaplan-Meier survival analysis. The number of postoperative IOP-lowering medications and occurrence of complications were also assessed. Mean preoperative IOP in the 120° group was 16.9 ± 7.6 mmHg, which significantly decreased to 10.9 ± 2.7 mmHg (p < 0.01) and 11.1 ± 3.1 mmHg (p = 0.01) at 12 and 24 months, respectively. The mean number of preoperative IOP-lowering medications significantly decreased from 2.8 ± 1.4 to 1.4 ± 1.4 (p < 0.01) at 24 months. Mean preoperative IOP in the 180° group was 17.1 ± 7.0 mmHg, which significantly decreased to 12.1 ± 3.2 mmHg (p = 0.02) and 12.9 ± 1.4 mmHg (p = 0.01) at 12 and 24 months, respectively. The mean number of preoperative IOP-lowering medications significantly decreased from 2.9 ± 1.2 to 1.4 ± 1.5 (p < 0.01) at 24 months. The probability of qualified success at 24 months in the 120° and 180° groups was 50.4% and 54.6%, respectively (p = 0.58). There was no difference observed for hyphema formation or IOP spikes. Surgical outcomes were not significantly different between the 120° and 180° incisions in Schlemm’s canal.

scholarly journals One-year outcomes of microhook trabeculotomy versus suture trabeculotomy ab interno

2021 ◽  
Author(s):  
Hiroshi Yokoyama ◽  
Masashi Takata ◽  
Fumi Gomi
Keyword(s):  
Survival Analysis ◽  
Clinical Success ◽  
Treatment Success ◽  
Medication Use ◽  
Glaucoma Surgery ◽  
Success Rates ◽  
Kaplan Meier ◽  
One Year ◽  
Ab Interno

Abstract Purpose To compare clinical success rates and reductions in intraocular pressure (IOP) and IOP-lowering medication use following suture trabeculotomy ab interno (S group) or microhook trabeculotomy (μ group). Methods This retrospective review collected data from S (n = 104, 122 eyes) and μ (n = 42, 47 eyes) groups who underwent treatment between June 1, 2016, and October 31, 2019, and had 12-month follow-up data including IOP, glaucoma medications, complications, and additional IOP-lowering procedures. The Kaplan–Meier survival analysis was used to evaluate treatment success rates defined as normal IOP (> 5 to ≤ 18 mm Hg), ≥ 20% reduction of IOP from baseline at two consecutive visits, and no further glaucoma surgery. Results Schlemm’s canal opening was longer in the S group than in the μ group (P < 0.0001). The Kaplan–Meier survival analysis of all eyes showed cumulative clinical success rates in S and µ groups were 71.1% and 61.7% (P = 0.230). The Kaplan–Meier survival analysis of eyes with preoperative IOP ≥ 21 mmHg showed cumulative clinical success rates in S and μ groups were 80.4% and 60.0% (P = 0.0192). There were no significant differences in postoperative IOP at 1, 3, and 6 months (S group, 14.9 ± 5.6, 14.6 ± 4.5, 14.6 ± 3.9 mmHg; μ group, 15.8 ± 5.9, 15.2 ± 4.4, 14.7 ± 3.7 mmHg; P = 0.364, 0.443, 0.823), but postoperative IOP was significantly lower in the S group at 12 months (S group, 14.1 ± 3.1 mmHg; μ group, 15.6 ± 4.1 mmHg; P = 0.0361). There were no significant differences in postoperative numbers of glaucoma medications at 1, 3, 6, and 12 months (S group, 1.8 ± 1.6, 1.8 ± 1.5, 2.0 ± 1.6, 1.8 ± 1.5; μ group, 2.0 ± 1.6, 2.0 ± 1.6, 2.1 ± 1.6, 2.2 ± 1.7; P = 0.699, 0.420, 0.737, 0.198). Conclusion S and µ group eyes achieved IOP reduction, but μ group eyes had lower clinical success rates among patients with high preoperative IOP at 12 months.

scholarly journals Brown Spiders’ Phospholipases-D with Potential Therapeutic Applications: Functional Assessment of Mutant Isoforms

Biomedicines ◽  
2021 ◽  
Vol 9 (3) ◽  
pp. 320
Author(s):  
Thaís Pereira da Silva ◽  
Fernando Jacomini de Castro ◽  
Larissa Vuitika ◽  
Nayanne Louise Costacurta Polli ◽  
Bruno César Antunes ◽  
...  
Keyword(s):  
Structural Changes ◽  
Capillary Permeability ◽  
Structural Data ◽  
Histopathological Analysis ◽  
Amino Acid Residues ◽  
Wild Type ◽  
Antigenic Properties ◽  
Harmful Effects

Phospholipases-D (PLDs) found in Loxosceles spiders’ venoms are responsible for the dermonecrosis triggered by envenomation. PLDs can also induce other local and systemic effects, such as massive inflammatory response, edema, and hemolysis. Recombinant PLDs reproduce all of the deleterious effects induced by Loxosceles whole venoms. Herein, wild type and mutant PLDs of two species involved in accidents—L. gaucho and L. laeta—were recombinantly expressed and characterized. The mutations are related to amino acid residues relevant for catalysis (H12-H47), magnesium ion coordination (E32-D34) and binding to phospholipid substrates (Y228 and Y228-Y229-W230). Circular dichroism and structural data demonstrated that the mutant isoforms did not undergo significant structural changes. Immunoassays showed that mutant PLDs exhibit conserved epitopes and kept their antigenic properties despite the mutations. Both in vitro (sphingomyelinase activity and hemolysis) and in vivo (capillary permeability, dermonecrotic activity, and histopathological analysis) assays showed that the PLDs with mutations H12-H47, E32-D34, and Y228-Y229-W230 displayed only residual activities. Results indicate that these mutant toxins are suitable for use as antigens to obtain neutralizing antisera with enhanced properties since they will be based on the most deleterious toxins in the venom and without causing severe harmful effects to the animals in which these sera are produced.

scholarly journals Autophosphorylation of DNA-PKCS regulates its dynamics at DNA double-strand breaks

2007 ◽  
Vol 177 (2) ◽  
pp. 219-229 ◽  
Author(s):  
Naoya Uematsu ◽  
Eric Weterings ◽  
Ken-ichi Yano ◽  
Keiko Morotomi-Yano ◽  
Burkhard Jakob ◽  
...  
Keyword(s):  
Kinase Activity ◽  
Laser System ◽  
Double Strand Breaks ◽  
Dependent Manner ◽  
Dna Double Strand Breaks ◽  
End Joining ◽  
Dsb Repair ◽  
Strand Breaks ◽  
Dna Ends

The DNA-dependent protein kinase catalytic subunit (DNA-PKCS) plays an important role during the repair of DNA double-strand breaks (DSBs). It is recruited to DNA ends in the early stages of the nonhomologous end-joining (NHEJ) process, which mediates DSB repair. To study DNA-PKCS recruitment in vivo, we used a laser system to introduce DSBs in a specified region of the cell nucleus. We show that DNA-PKCS accumulates at DSB sites in a Ku80-dependent manner, and that neither the kinase activity nor the phosphorylation status of DNA-PKCS influences its initial accumulation. However, impairment of both of these functions results in deficient DSB repair and the maintained presence of DNA-PKCS at unrepaired DSBs. The use of photobleaching techniques allowed us to determine that the kinase activity and phosphorylation status of DNA-PKCS influence the stability of its binding to DNA ends. We suggest a model in which DNA-PKCS phosphorylation/autophosphorylation facilitates NHEJ by destabilizing the interaction of DNA-PKCS with the DNA ends.

scholarly journals Metabolism of rat bone proteoglycans in vivo

1983 ◽  
Vol 216 (3) ◽  
pp. 589-596 ◽  
Author(s):  
C W Prince ◽  
F Rahemtulla ◽  
W T Butler
Keyword(s):  
Extraction Procedure ◽  
Total Radioactivity ◽  
Mineralized Tissue ◽  
Guanidinium Chloride ◽  
Disaccharide Composition ◽  
A Minor ◽  
Papain Digestion ◽  
Rat Bone

Former evaluations of the role of proteoglycans in mineralization have neglected to address the possibility that the metabolism of proteoglycans may be of significance in this regard. This problem was studied by using radiolabeling in vivo of rat calvaria with [35Sulphate for 2-72 h and a sequential extraction procedure to yield two pools of newly synthesized proteoglycans: one obtained from non-mineralized tissue by extraction with guanidinium chloride (GdmCl) and another obtained only after demineralization with EDTA. Total radioactivity in calvaria was maximal after 12 h of incorporation, but by 36 h had declined to a level that was about 55-65% of maximum. Radioactivity in the GdmCl extract declined steadily after 12 h, whereas that in the EDTA extract remained constant until 36 h, when it began to increase. Each extract contained a minor proteoglycan that eluted at the void volume (Vo) of a Sepharose CL-6B column. Unlike in the EDTA extract, this proteoglycan gradually disappeared from the GdmCl extract. Each extract also contained a major, smaller proteoglycan, with a Kav. of 0.24 and 0.36 in the GdmCl and EDTA extracts respectively. Papain digestion of each extract yielded glycosaminoglycan chains with Kav. values of 0.32 and 0.50 on CL-6B in the GdmCl and EDTA extracts respectively. Digestion of each extract with chondroitinase ABC and chondroitinase AC showed that the glycosaminoglycans were of similar disaccharide composition, with about 85% being 4-sulphated and the remainder 6-sulphated and/or iduronic acid-containing. These data suggest that about 45% of the newly synthesized proteoglycans are removed from the tissue during the course of mineralization.

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