E3MPH16: An efficient endosomolytic peptide for intracellular protein delivery.

To facilitate the introduction of proteins, such as antibodies, into cells, a variety of delivery peptides have been engineered. These peptides are typically highly cationic and somewhat hydrophobic, enabling cytosolic protein delivery at the cost of causing cell damage by rupturing membranes. This balance between delivery effectiveness and cytotoxicity presents obstacles for their real-world use. To tackle this problem, we designed a new endosome-disruptive cytosolic delivery peptide, E3MPH16, inspired by mastoparan X (MP). E3MPH16 was engineered to incorporate three Glu (E3) and 16 His (H16) residues at the N- and C-termini of MP, respectively. The negative charges of E3 substantially mitigate the cell-surface damage induced by MP. The H16 segment is known to enhance cell-surface adsorption and endocytic uptake of the associated molecules. With these modifications, E3MPH16 was successfully trapped within endosomes. The acidification of endosomes is expected to protonate the side chains of E3 and H16, enabling E3MPH16 to rupture endosomal membranes. As a result, nearly 100% of cells achieved cytosolic delivery of a model biomacromolecule, Alexa Fluor 488-labeled dextran (10 kDa), via endosomal escape by co-incubation with E3MPH16. The delivery process also suggested the involvement of macropinocytosis and caveolae-mediated endocytosis. With the assistance of E3MPH16, Cre recombinase and anti-Ras-IgG delivered into HEK293 cells and HT1080 cells enabled gene recombination and inhibited cell proliferation, respectively. The potential for in vivo application of this intracellular delivery method was further validated by topically injecting the green fluorescent protein fused with a nuclear localization signal (NLS-GFP) along with E3MPH16 into Colon-26 tumor xenografts in mice.

One-year recurrence rate of Graves ophthalmopathy presenting as diplopia in the primary position after varied doses of intravenous methylprednisolone followed by oral prednisolone with dosing based on the magnetic resonance imaging findings.

PURPOSE: To reveal the recurrence rate of Graves ophthalmopathy (GO) presenting as diplopia in the primary position for 1 year after varied doses of intravenous methylprednisolone (IVMP) followed by oral prednisolone, with dosing based on the magnetic resonance imaging (MRI) findings. STUDY DESIGN: Retrospective study. METHODS: We analyzed the medical charts of 25 patients who were diagnosed with new-onset GO and who received treatment for diplopia in the primary position at our hospital. Treatment consisted of MRI-determined varied doses of IVMP followed by oral prednisolone. If the MRI findings showed deterioration or were unchanged after 6 g of IVMP, 3 g of IVMP was added for further treatment. Simple and multiple linear regression analyses were performed to reveal the associations between the independent variables and the dependent variable, defined as recurrence. RESULTS: The mean patient age (± standard deviation) was 61.3 ± 11.3 years. The female to male ratio was 15:10. Twenty-one of the 25 patients received a total of 6 g of IVMP, whilst the remaining 4 patients received a total of 9 g of IVMP. In 5 patients (20%), the GO recurred within 1 year of IVMP administration. Simple and multiple linear regression analyses showed that the MRI findings after 6 g of IVMP affected recurrence (P < .05). CONCLUSION: This study showed that in 20% of patients, GO recurred within 1 year of administration of varied doses of IVMP, with the dosing based on the MRI findings. Furthermore, assessment of inflammation by use of MRI after 6 g of IVMP has a potential role in predicting recurrence.

Posterior microphthalmos with achievement of good visual acuity and disappearance of papillomacular retinal folds: a case report.

BACKGROUND: Posterior microphthalmos (PM) is a rare condition with poor visual prognosis even after amblyopia treatment. We report a case of PM with achievement of good visual acuity and disappearance of papillomacular retinal folds (PFs) over a period of 7 years. CASE PRESENTATION: A girl aged 3 years and 5 months was referred to our hospital, after poor visual acuity was identified at a medical checkup for 3-year-olds. She had severe spherical hyperopia: + 17.25 D in the right eye (RE) and + 18 D in the left eye (LE). Her corrected visual acuity was 20/200 in the RE and 20/250 in the LE. PFs were observed in both eyes on optical coherence tomography (OCT), and the diagnosis of PM was made based on the normal corneal diameter and anterior chamber depth. During the course of the disease, a gradual decrease in the height of the PFs was observed on OCT. The corrected visual acuity at age 10 years was 20/20 in the RE and 20/25 in the LE. CONCLUSIONS: The visual prognosis of PM is poor, and only one case with good visual acuity has been reported in the literature. The patient in the present case not only developed good visual acuity, but also showed improvement in macular morphology, which was not noted in previous reports. Early diagnosis of PM and early amblyopia treatment is important for the visual development in PM.

Hyperopic anisometropia with a shorter axial length ipsilateral to the ptotic eye in children with congenital ptosis.

BACKGROUND: To investigate the clinical characteristics of children with congenital ptosis, with particular attention given to the incidence of anisometropia, and the difference in axial length (AL) between the right and left eyes. METHODS: The medical charts of 55 patients with congenital ptosis at Niigata University Medical and Dental Hospital were retrospectively analyzed. Clinical characteristics, including age, cycloplegic refraction, AL, and the presence of amblyopia and its causes were analyzed. RESULTS: Age at the initial visit was 16 ± 20 (mean ± standard deviation, the same applies below) months. Of the 49 patients whose cycloplegic refraction was measured, hyperopic anisometropia, defined as ≥ one-diopter difference in spherical equivalent (SE), was observed in 1/11, 9/27 and 5/11 patients with bilateral, right, and left ptosis, respectively. Among 14/38 patients with hyperopic anisometropia involving unilateral ptosis, 13 demonstrated a larger SE in the ptotic eye than in the non-ptotic eye. The inter-eye difference in AL (AL of the ptotic eye minus that of the non-ptotic eye) in six patients with unilateral ptosis and hyperopic anisometropia ipsilateral to the ptotic eye (-0.29 ± 0.40 mm) was significantly smaller than that in three patients with unilateral ptosis and no hyperopic anisometropia (0.38 ± 0.29 mm). CONCLUSIONS: At our institute, children with congenital ptosis had a high incidence of hyperopic anisometropia ipsilateral to the ptotic eye. Furthermore, this condition was associated with a shorter axial length. These results indicate that refractive correction for hyperopic anisometropia is important for proper visual development in children with congenital ptosis.

The Prevalence of Brain Abnormalities in Japanese Patients with Optic Nerve Hypoplasia.

The purpose of this study was to investigate the clinical characteristics of Japanese patients with optic nerve hypoplasia (ONH), with particular attention to the prevalence of brain abnormalities. We retrospectively analysed the medical charts of 16 patients who were diagnosed with ONH and who underwent magnetic resonance imaging (MRI) at Niigata University Medical and Dental Hospital. We recorded the age, sex, laterality, initial eye and visual symptoms, best-corrected visual acuity, and brain abnormalities on MRI (excluding ONH). The median age at the first visit to the Ophthalmology Clinic was 2.4 years old. Four patients were male and 12 were female. ONH was bilateral in 11 patients and unilateral in five. Best-corrected visual acuity ranged from no light perception to 20/20. Seven patients (43.8%) had brain abnormalities including agenesis of the septum pellucidum, pituitary gland hypofunction, cerebral dysplasia, and West syndrome. Five of these seven patients had general manifestations since the neonatal or infantile period. Our study revealed the prevalence of brain abnormalities associated with optic nerve hypoplasia in Japanese patients at a single institute. Because two of 11 patients had no general manifestations since the neonatal or infantile period but demonstrated brain abnormalities, MRI should be performed to investigate all patients with ONH.

Visual outcome of aquaporin-4 antibody-positive optic neuritis with maintenance therapy.

PURPOSE: To assess the effect of maintenance therapy on visual outcomes in preventing recurrences one year after first onset in patients with aquaporin-4 antibody (AQP4Ab)-positive optic neuritis. STUDY DESIGN: Retrospective study. METHODS: The medical charts of 56 patients with optic neuritis (22 with AQP4Ab-positive and 34 with AQP4Ab-negative) at Niigata University Medical and Dental Hospital were retrospectively analyzed. Clinical characteristics, including visual acuity and number of recurrences one year after first onset, were compared among patients who were AQP4Ab-positivie with and those without maintenance therapy such as oral prednisolone and azathioprine, as well as those who were AQP4Ab-negative. RESULTS: The mean ages were 49.3 and 45.2 years in the AQP4Ab-positive and the AQP4Ab-negative groups. The female to male ratio was 21:1 and 18:16 in the two groups, respectively. Multiple between-group comparison showed a statistically significant difference in visual acuity one year after first onset between the AQP4Ab-positive without maintenance therapy group and the AQP4Ab-negative group (0.05 (median, same applies below) vs. 1.0, p < 0.01). There was also a statistically significant difference in the number of recurrences in the year after first onset between the AQP4Ab-positive with and without maintenance therapy groups (1 vs. 0, p < 0.01). CONCLUSION: This study demonstrates that patients with AQP4Ab-positive optic neuritis without maintenance therapy had the poorest visual acuity and the most recurrences one year after first onset. These results indicate that reducing the number of recurrences with maintenance therapy could improve the visual outcomes in patients with AQP4Ab-positive optic neuritis.