Single-Pass Mikrokeratome and Anterior Chamber Pressurizer for the Ultrathin Descemet-Stripping Automated Endothelial Keratoplasty Graft Preparation.

PURPOSE: To compare the reliability of microkeratome dissection with the anterior chamber pressurizer (ACP) system versus conventional pressurization for ultrathin Descemet-stripping automated endothelial keratoplasty (UT-DSAEK) graft preparation. METHODS: A retrospective review of a consecutive series of dissected donor corneas processed at Pavia Eye Bank for UT-DSAEK surgery was performed. Grafts were prepared through single-pass microkeratome dissection with artificial anterior chamber internal pressure regulation through either ACP or the conventional method using a water column with tube clamp. The target central graft thickness (CGT) was ≤100 µm. Cutting predictability was determined as the difference between the microkeratome head size and the thickness actually removed. Graft regularity was investigated as central-to-peripheral thickness increase, central-to-peripheral (CP) ratio, and graft thickness uniformity. Thickness was measured with anterior segment optical coherence tomography (horizontal and vertical meridians). RESULTS: Of the 265 UT-DSAEK grafts, ACP achieved the target "CGT ≤ 100 µm" in 87 of 120 (72.5%), whereas the conventional technique achieved the same in 85 of 145 (58.6%) (P = 0.018). ACP predictability was -3.9 µm (SD: 2.3), whereas predictability of the conventional technique was -54.6 µm (SD: 3.7) (P < 0.001). Thickness increased similarly (P = 0.212); CP ratio was better with ACP for only 2 mm diameter (P = 0.001); graft thickness uniformity was comparable (P > 0.05). CONCLUSIONS: Compared with conventional pressurization, ACP improved microkeratome-assisted preparation reliability of UT-DSAEK grafts, achieving CGT ≤ 100 µm with significantly higher frequency (P = 0.018) and predictability (P < 0.001). ACP improved CP ratio only at 2 mm (P = 0.001); for other graft thickness, the 2 methods proved equivalent.

A case report of pediatric neurotrophic keratopathy in pontine tegmental cap dysplasia treated with cenegermin eye drops.

RATIONALE: To report the management of recalcitrant neurotrophic keratopathy in a pediatric patient affected by pontine tegmental cap dysplasia (PTCD) using topical human recombinant nerve growth factor (hrNGF, Cenegermin 20 µg/ml). To the best of our knowledge the present case is one of the few described in patients with congenital NK treated with Cenegermin, and the first in a patient affected by PTCD. PATIENT CONCERNS: A 9-year-old patient, affected by PTCD with bilateral cranial nerve V1 and VIII palsies, was referred to our hospital for visual disturbances and redness of the right eye due to persistent neurotrophic epithelial defect. The patient presented marked developmental delay, ataxia, bilateral hypoacusia, and bilateral corneal severe hypoaesthesia. Ocular history revealed multiple treatments in order to treat neurotrophic ulcer in the left eye. Four years later, he developed a persistent epithelial defect with corneal anesthesia in the right eye. DIAGNOSES: The impaired trigeminal nerve function, due to the underlying congenital disease, led to the development of moderate NK (stage II) in the right eye and a mild NK (stage I) in the left eye. INTERVENTIONS: Cenegermin 20 µg/ml eye-drop was administered in both eyes. Treatment was continued for 8 weeks. The patient was assessed after 4 and 8 week of treatment. At each follow-up visit, treatment efficacy and adverse events were evaluated. OUTCOMES: The use of Cenegermin eye drops facilitated the remarkable resolution of the neurotrophic keratopathy and the improvement of corneal sensitivity in both eyes. No local or systemic adverse events were observed. LESSONS: Topical Cenegermin 20 µg/ml was well-tolerated and may represent a valuable therapeutic option in the management of pediatric neurotrophic keratopathy.

Mass spectrometric strategies for the identification and characterization of human serum albumin covalently adducted by amoxicillin: ex vivo studies.

This study addresses the detection and characterization of the modification of human serum albumin (HSA) by amoxicillin (AX) in ex vivo samples from healthy subjects under oral amoxicillin administration (acute intake of 1 g every 8 h for 48 h). To reach this goal, we used an analytical strategy based on targeted and untargeted mass spectrometric approaches. Plasma samples withdrawn before AX oral intake represented the negative control samples to test the method selectivity, whereas HSA incubated in vitro with AX was the positive control. Different MS strategies were developed, particularly (1) multiple reaction monitoring (MRM) and precursor ion scan (PIS) using a HPLC system coupled to a triple quadrupole MS analyzer and (2) a dedicated data-dependent scan and a customized targeted MS/MS analysis carried out using a nano-LC system coupled to a high-resolution MS system (LTQ Orbitrap XL). Lys 190 was identified as the only modification site of HSA in the ex vivo samples. The AX adduct was identified and fully characterized by complementary targeted approaches based on triple quadrupole (MRM mode) and orbitrap (SIC mode) mass analyzers. The SIC mode also permitted the relative amount of AX-adducted HSA to be measured, ranging from 1 to 2% (6-12 µM) at 24 and 48 h after the oral intake. No adduct in any ex vivo sample was identified by the untargeted methods (PIS and data-dependent scan mode analysis). The results on one hand indicate that MS, in particular high-resolution MS, analysis represents a suitable analytical tool for the identification/characterization of covalently modified proteins/peptides; on the other hand, they give deeper insight into AX-induced protein haptenation, which is required to better understand the mechanisms involved in AX-elicited allergic reactions.