Clinical utility of the KAMRA corneal inlay

Shehzad Anjam Naroo, Paramdeep Singh Bilkhu

Ophthalmic Research Group, School of Life & Health Sciences, Aston University, Birmingham, UK

Abstract: The treatment of presbyopia has been the focus of much scientific and clinical research over recent years, not least due to an increasingly aging population but also the desire for spectacle independence. Many lens and nonlens-based approaches have been investigated, and with advances in biomaterials and improved surgical methods, removable corneal inlays have been developed. One such development is the KAMRA™ inlay where a small entrance pupil is exploited to create a pinhole-type effect that increases the depth of focus and enables improvement in near visual acuity. Short- and long-term clinical studies have all reported significant improvement in near and intermediate vision compared to preoperative measures following monocular implantation (nondominant eye), with a large proportion of patients achieving Jaeger (J) 2 to J1 (~0.00 logMAR to ~0.10 logMAR) at the final follow-up. Although distance acuity is reduced slightly in the treated eye, binocular visual acuity and function remain very good (mean 0.10 logMAR or better). The safety of the inlay is well established and easily removable, and although some patients have developed corneal changes, these are clinically insignificant and the incidence appears to reduce markedly with advancements in KAMRA design, implantation technique, and femtosecond laser technology. This review aims to summarize the currently published peer-reviewed studies on the safety and efficacy of the KAMRA inlay and discusses the surgical and clinical outcomes with respect to the patient's visual function.

Keywords: presbyopia, refractive surgery, implants, cornea


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Time of maximum cycloplegia after instillation of cyclopentolate 1% in children with brown irises

Sittikorn Laojaroenwanit, Vimontip Layanun, Pokpong Praneeprachachon, Parnchat Pukrushpan

Department of Ophthalmology, Faculty of Medicine, Chulalongkorn University, Bangkok, Thailand

Purpose: We aimed to 1) determine the time of maximum cycloplegia after instillation of cyclopentolate 1% in children with brown irises, 2) evaluate the correlation between the pupillary reaction and time of maximum cycloplegia, and 3) identify any side effects of the medication.
Patients and methods: This was a prospective analytical study involving children aged 5 to 14 years who were attending refraction clinic. Cyclopentolate 1% was instilled three times at 10-minute intervals. The spherical equivalent, pupillary reaction, and pupillary diameter were recorded before the first drop and nine times after the last drop at 10-minute intervals. Side effects were assessed. Time of maximum cycloplegia was determined from the time point at which the 95% confidence interval of the differences between the mean spherical equivalent at each point and its final value at 110 minutes was reached and remained within the equivalence limit (±0.25 D).
Results: Sixty children were enrolled in this study. Their mean age was 9.8 years (range: 5–14 years). Time of maximum cycloplegia was reached at 30 minutes after the first instillation of cyclopentolate. A poor correlation was observed between the pupillary reaction and the time of maximum cycloplegia (r=−0.07). The mean pupillary diameter at 30 minutes was 3.7±1.3 mm, and further dilation occurred thereafter. No side effects were observed.
Conclusion: In most children, maximum cycloplegia was reached 30 minutes after the first instillation of cyclopentolate. The absence of a pupillary reaction should not be used as an indicator of maximum cycloplegia.

Keywords: refraction, cyclopentolate, cycloplegia, brown irises, cycloplegic refraction


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Ranibizumab in monotherapy and combined with photodynamic therapy for retinal angiomatous proliferation

Luis Arias,^1–3 Francisco Gómez-Ulla,^2–4 José M Ruiz-Moreno<sup>2,3,5

1</sup>Ophthalmology Department, Bellvitge University Hospital, C/Feixa Llarga, L'Hospitalet de Llobregat, Barcelona, ²Spanish Vitreoretinal Society (SERV), C/Xosé Chao Rego, Santiago de Compostela, ³RETICS OFTARED, Institute of Health Carlos III, C/Sinesio Delgado, Madrid, ⁴Gómez-Ulla Eye Institute, Santiago de Compostela, ⁵Department of Ophthalmology, Albacete University Hospital, Avenida de Almansa s/n, Albacete, Spain

Purpose: To compare the effects of intravitreal ranibizumab in monotherapy (group A) and combined with photodynamic therapy (PDT) with verteporfin (group B) in retinal angiomatous proliferation (RAP) treatment.
Methods: This was a multicentric, prospective, randomized clinical study conducted with parallel groups. The study eye in both groups received ranibizumab on days 1, 30, and 60 (loading dose); group B received PDT additionally on day 1. Early Treatment Diabetic Retinopathy Study (ETDRS) visual acuity (VA) testing and optical coherence tomography were performed monthly, and fluorescein angiography and indocyanine green angiography were performed quarterly. Retreatment criteria were leakage in fluorescein angiography or indocyanine green angiography, mean foveal thickness increase ≥100 µm, or VA decrease ≥5 letters.
Results: Twenty patients were recruited (ten patients in each group). Six eyes had previous treatment (three eyes in group A and three eyes in group B), so only 14 eyes were naïve. At 12-month follow-up, mean VA improved +1.5 letters in group A and +5.6 letters in group B (analysis of variance test; P>0.05). Two patients (20%) in both groups gained ≥15 letters (chi-square test; P>0.05). Mean changes in greatest linear dimension and in foveal thickness were not statistically significant between groups of treatment (analysis of variance test; P>0.05). Mean retreatments per patient were 1.8 (group A) and 0.9 (group B) (Mann–Whitney U-test; P>0.05). One patient died due to underlying disease not related to study medication.
Conclusion: Intravitreal ranibizumab administered in monotherapy or combined with PDT was efficacious in terms of VA stabilization in patients with RAP.

Keywords: age-related macular degeneration, intravitreal injection, photodynamic therapy, ranibizumab, retinal angiomatous proliferation, verteporfin


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Segmentation and removal of fibrovascular membranes with high-speed 23 G transconjunctival sutureless vitrectomy, in severe proliferative diabetic retinopathy

Erkan Celik,¹ Ozkan Sever,² Fatih Horozoglu,² Ates Yanyali<sup>3

1</sup>Sakarya University Medical Education and Research Hospital, Sakarya, ²Namik Kemal University, School of Medicine, Tekirdag, ³Haydarpasa Numune Medical Education and Research Hospital, Istanbul, Turkey

Aim: To evaluate the effectiveness and safety of high-speed (5,000 cuts per minute) 23 G transconjunctival sutureless vitrectomy (TSV) in severe diabetic fibrovascular proliferation (DFVP).
Patients and methods: In this retrospective consecutive case series, patients who underwent 23 G TSV for severe DFVP between October 2011 and March 2014 at our institution were evaluated. 23 G TSV was performed with a high-speed (5,000 cuts per minute) cutter without a chandelier light.
Results: The mean follow-up period was 8 months (range: 4–23 months). Of the 27 eyes of 27 patients, 14 eyes (52%) underwent concomitant phacoemulsification with posterior chamber intraocular lens implantation, nine eyes (33%) were pseudophakic, and four eyes were phakic (15%). DFVP was removed with ease in all, and visual acuity was improved in 18 (67%) eyes. Iatrogenic retinal tear was observed in four eyes (15%) and treated successfully during surgery. Suture placement to a single sclerotomy was performed in eight eyes (30%). Postoperative intraocular hemorrhage was observed in five eyes (18%). Cataract formation was observed in two of the four phakic eyes. Three (11%) patients had postoperative intraocular pressure rise. Postoperative hypotony (≤6 mmHg) and endophthalmitis were not observed in any eye.
Conclusion: The segmentation and removal of fibrovascular membranes with high-speed 23 G TSV seems to be a safe and easy method in severe diabetic eye disease.

Keywords: diabetic fibrovascular proliferation, transconjunctival sutureless vitrectomy, high speed


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Neurofibromatosis: an update of ophthalmic characteristics and applications of optical coherence tomography

Barmak Abdolrahimzadeh,¹ Domenica Carmen Piraino,² Giorgio Albanese,² Filippo Cruciani,² Siavash Rahimi<sup>3

1</sup>Polimed Beltramelli Medical Center, Rome, Italy; ²Section of Ophthalmology, Department of Sense Organs, University of Rome "Sapienza", Rome, Italy; ³Pathology Centre, Queen Alexandra Hospital, Portsmouth, UK

Abstract: Neurofibromatosis (NF) is a multisystem disorder and tumor predisposition syndrome caused by genetic mutation on chromosome 17-17q11.2 in NF type 1 (NF1), and on chromosome 22-22q12.2 in NF type 2. The disorder is characterized by considerable heterogeneity of clinical expression. NF1 is the form with the most characteristic ocular manifestations. Lisch nodules of the iris are among the well-known diagnostic criteria for the disease. Glaucoma and associated globe enlargement have been described in a significant proportion of patients with NF1 and orbital–facial involvement. Optic nerve glioma may cause strabismus and proptosis, and palpebral neurofibroma may reach considerable size and occasionally show malignant transformation. Near infrared reflectance has greatly contributed to enhancing our knowledge on choroidal alterations in NF1. Indeed, some authors have proposed to include these among the diagnostic criteria. Optical coherence tomography has given new insight on retinal alterations and is a noninvasive tool in the management of optic nerve gliomas in children. Ocular manifestations in NF type 2 can range from early-onset cataracts in up to 80% of cases to optic nerve hamartomas and combined pigment epithelial and retinal hamartomas.

Keywords: neurofibromatosis, ophthalmic, optical coherence tomography, infrared reflectance, choroideal nodules, Lisch nodules

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Tear volume estimation using a modified Schirmer test: a randomized, multicenter, double-blind trial comparing 3% diquafosol ophthalmic solution and artificial tears in dry eye patients

Hideki Miyake,¹ Yuri Kawano,² Hiroshi Tanaka,² Akihiro Iwata,³ Takahiro Imanaka,¹ Masatsugu Nakamura<sup>1

1</sup>Ophthalmic Disease Area Strategy Department, ²Clinical Operations Department, ³Data Science Department, R&D Division, Santen Pharmaceutical Co., Ltd. Osaka, Japan

Purpose: We aimed to evaluate the feasibility of using a modified Schirmer test to determine the increase in tear volume after administration of 3% diquafosol ophthalmic solution (diquafosol 3%) in dry eye patients.
Patients and methods: A randomized, multicenter, prospective, double-blind clinical study recruited 50 qualified subjects. They received diquafosol 3% in one eye and artificial tears in the other eye. The study protocol comprised a screening and treatment procedure completed within 1 day. The Schirmer test was performed on closed eyes three times a day. The primary efficacy end points were the second Schirmer test scores 10 minutes after the single dose. Secondary end points were the third Schirmer test scores 3 hours and 40 minutes after the single dose and the symptom scores prior to the second and third Schirmer tests.
Results: According to the Schirmer test, 10 minutes after administration, diquafosol 3% significantly increased tear volume compared to artificial tears. Diquafosol 3% and artificial tears both showed significant improvements in the symptom scores compared to baseline. However, there was no significant difference in the symptoms score between diquafosol 3% and artificial tears.
Conclusion: The modified Schirmer test can detect a minute change in tear volume in dry eye patients. These findings will be useful in the diagnosis of dry eye, assessment of treatment benefits in daily clinical practice, and the development of possible tear-secreting compounds for dry eye.

Keywords: P2Y2, efficacy, Diquas^®


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Ocular manifestations of Sturge–Weber syndrome: pathogenesis, diagnosis, and management

Flavio Mantelli,¹ Alice Bruscolini,² Maurizio La Cava,² Solmaz Abdolrahimzadeh,² Alessandro Lambiase<sup>2

1</sup>Department of Biology, College of Science and Technology, Temple University, Philadelphia, PA, USA; ²Department of Sense Organs, Section of Ophthalmology, University of Rome "Sapienza", Rome, Italy

Abstract: Sturge–Weber syndrome has been included in the group of phakomatoses that is characterized by hamartomas involving the brain, skin, and eyes. The characteristic facial port-wine stain, involving the first branch of the trigeminal nerve and the embryonic vasculature distribution in this area, leads to several ocular complications of the anterior segment and can involve the eyelids and conjunctiva. The posterior segment of the eyes is also affected with diffuse choroidal hemangiomas. However, the most frequent ocular comorbidity is glaucoma with a prevalence rate ranging from 30%–70%. Glaucoma is related to anterior chamber malformations, high episcleral venous pressure (EVP), and changes in ocular hemodynamics. Glaucoma can be diagnosed at birth, but the disease can also develop during childhood and in adults. The management of glaucoma in Sturge–Weber syndrome patients is particularly challenging because of early onset, frequently associated severe visual field impairment at the time of diagnosis, and unresponsiveness to standard treatment. Several surgical approaches have been proposed, but long-term prognosis for both intraocular pressure control and visual function remains unsatisfactory in these patients. Choroidal hemangiomas may also lead to visual impairment thorough exudative retinal detachment and macular edema. Treatment of exudative hemangioma complications is aimed at destructing the tumor or decreasing tumor leakage.

Keywords: Sturge–Weber syndrome, glaucoma, choroidal hemangiomas, port-wine stain, congenital disease, glaucoma surgery

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The IMPACT study: a prospective evaluation of the effects of cyclosporine ophthalmic emulsion 0.05% on ocular surface staining and visual performance in patients with dry eye

Karl G Stonecipher,^1,2 Gail L Torkildsen,³ George W Ousler III,⁴ Scot Morris,⁵ Linda Villanueva,⁶ David A Hollander<sup>6

1</sup>Department of Ophthalmology, University of North Carolina, Chapel Hill, ²TLC Laser Eye Centers, Greensboro, NC, ³Andover Eye Associates, ⁴Ora, Inc., Andover, MA, ⁵Eye Consultants of Colorado, Conifer, CO, ⁶Allergan plc, Irvine, CA, USA

Objective: The aim of this study was to evaluate the effects of cyclosporine ophthalmic emulsion 0.05% on ocular surface staining and visual performance in patients with dry eye.
Methods: This was a single-center, 6-month, open-label, Phase IV study. Patients with bilateral dry eye disease and a symptom score of ≥2 on the Ocular Discomfort and 4-Symptom Questionnaire, an Ocular Surface Disease Index score of >12, at least one eye with Schirmer's score <10 mm/5 minutes, and central corneal staining graded as ≥2 on the Ora Calibra™ Corneal and Conjunctival Staining Scale were enrolled. Cyclosporine ophthalmic emulsion 0.05% (Restasis^®) was instilled twice daily in each eye. The primary efficacy endpoints were ocular surface staining and visual function at 6 months. Secondary outcome measures included Schirmer's test, tear film breakup time, symptoms, and adverse events.
Results: A total of 40 patients with the mean age of 59.4 years (range, 40–78 years) were enrolled; 35 (87.5%) were female and 37 (92.5%) completed the study. At 6 months, inferior corneal, central corneal, total corneal, and total ocular surface fluorescein staining were significantly improved from baseline in both eyes (P<0.001). Patient responses on the Ocular Surface Disease Index showed significant improvement in blurred vision and visual function related to reading, driving at night, working with a computer or bank machine, and watching television (P≤0.041). At 6 months, 35.1% of patients achieved ≥5 mm improvement and 18.9% achieved ≥10 mm improvement in the average eye Schirmer score. Mean tear film breakup time improved by >50% in both eyes (P<0.001). Patients reported significant improvement in ocular discomfort and dry eye symptoms (P<0.001). No patients discontinued treatment because of stinging or any other ocular adverse event.
Conclusion: Dry eye patients with difficulties with day-to-day visual function demonstrated improvement in both signs and symptoms of dry eye and reported improved visual function after 6 months of treatment with cyclosporine ophthalmic emulsion 0.05%.

Keywords: corneal staining, dry eye disease, Ocular Surface Disease Index, ocular surface staining, visual function, clinical trial

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An exploratory study evaluating the effects of macular carotenoid supplementation in various retinal diseases

Roxanne Crosby-Nwaobi, Philip Hykin, Tunde Peto, Sobha Sivaprasad

NIHR Clinical Research Facility, NIHR Moorfields Biomedical Research Centre, London, UK

Purpose: The aim of this study was to assess the impact of daily oral supplementation with Macushield (10 mg/d meso-zeaxanthin, 10 mg/d lutein, and 2 mg/d zeaxanthin) on eye health in patients with retinal diseases by assessing the macular pigment (MP) profile, the visual function, and the quality of life.
Methods: Fifty-one patients with various retinal diseases were supplemented daily and followed up for 6 months. The MP optical density was measured using the customized heterochromatic flicker photometry and dual-wavelength autofluorescence. Visual function was evaluated by assessing the change in best corrected visual acuity, contrast sensitivity, and glare sensitivity in mesopic and photopic conditions. Vision-related and general quality of life changes were determined using the National Eye Insititute- Visual Function Questionnaire-25 (NEI-VFQ-25) and EuroQoL-5 dimension questionnaires.
Results: A statistically significant increase in the MP optical density was observed using the dual-wavelength autofluorescence (P=0.04) but not with the customized heterochromatic flicker photometry. Statistically significant (P<0.05) improvements in glare sensitivity in low and medium spatial frequencies were observed at 3 months and 6 months. Ceiling effects confounded other visual function tests and quality of life changes.
Conclusion: Supplementation with the three carotenoids enhances certain aspects of visual performance in retinal diseases.

Keywords: macular pigment optical density, diabetes, central serous retinopathy, age-related macular degeneration


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Pre- and postsynaptic effects of brimonidine on isolated rabbit iris dilator muscles

Sonoko Tatsui,¹ Hitoshi Ishikawa,² Kimiya Shimizu,¹ Kimiyo Mashimo<sup>1

1</sup>Department of Ophthalmology, School of Medicine, Kitasato University, ²Department of Orthoptics and Visual Sciences, School of Allied Health Sciences, Kitasato University, Sagamihara, Japan

Purpose: Brimonidine is an imidazoline compound used for the treatment of glaucoma, but having very little effect on pupil diameter. Like para-aminoclonidine, most imidazoline compounds interact with postsynaptic α-adrenoceptors and cause pupil dilatation. Therefore, as part of an investigation of the mechanism of action of brimonidine on pupil diameter, the present study was initiated to measure, in vitro, the relative potency of brimonidine on the pre- and postsynaptic α-adrenoceptors of rabbit iris dilator muscle.
Methods: The contractile activity of brimonidine and its effect on twitch contraction evoked by electrical field stimulation were studied in isolated rabbit iris dilator muscles by isometric tension recording.
Results: Brimonidine significantly inhibited the twitch contraction of the dilator muscle caused by field stimulation, without affecting the response to exogenously applied phenylephrine. Compared to phenylephrine, brimonidine caused only a small contractile response with % maximum contraction values of<10%.
Conclusion: These results suggest that brimonidine may act on nerve endings to inhibit adrenergic neurotransmission with very little effect on postsynaptic α-adrenoceptors. This may indicate that brimonidine reduced the pupil diameter just a little, thus improving night vision.

Keywords: brimonidine, rabbit iris dilator, electrical field stimulation, presynaptic α₂-adrenoceptor, postsynaptic α₁-adrenoceptor, imidazolin


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Comparison of intraocular pressure adjusted by central corneal thickness or corneal biomechanical properties as measured in glaucomatous eyes using noncontact tonometers and the Goldmann applanation tonometer

Kiyoshi Yaoeda,^1,2 Atsushi Fukushima,¹ Motohiro Shirakashi,³ Takeo Fukuchi<sup>2

1</sup>Yaoeda Eye Clinic, Nagaoka, ²Division of Ophthalmology and Visual Sciences, Niigata University Graduate School of Medical and Dental Sciences, ³Kido Eye Clinic, Niigata, Japan

Purpose: To investigate the correlation coefficients between intraocular pressure (IOP) before and after adjusting for central corneal thickness (CCT) and corneal biomechanical properties.
Patients and methods: A total of 218 eyes of 218 patients with primary open-angle glaucoma (mean age =71.5 years; mean spherical equivalent =-0.51 D; mean deviation determined by Humphrey visual field analyzer =-3.22 dB) were included in this study. The tIOP and tIOPCCT, which were adjusted by the CCT (with tIOP meaning IOP not adjusted by CCT, as determined using the CT-1P; and tIOPCCT meaning IOP adjusted by CCT, as determined using the CT-1P), were determined using a noncontact tonometer. The IOPg and IOPCCT, which were adjusted by CCT, and IOPcc adjusted by corneal biomechanical properties were determined using a Reichert 7CR (with IOPg meaning IOP not adjusted by CCT or corneal biomechanical properties, as determined using the Reichert 7CR; IOPCCT meaning IOP adjusted by CCT, as determined using the Reichert 7CR; and IOPcc meaning IOP adjusted by corneal biomechanical properties, as determined using the Reichert 7CR). The GT and GTCCT adjusted by CCT were determined using a Goldmann applanation tonometer (with GT meaning IOP not adjusted by CCT, as determined using the Goldmann applanation tonometer; and with GTCCT meaning IOP adjusted by CCT, as determined using the GAT). Pearson's correlation coefficients among the IOPs were calculated and compared. P-values <0.05 were considered as statistically significant.
Results: The tIOP, tIOPCCT, IOPg, IOPCCT, IOPcc, GT, and GTCCT were 14.8±2.5, 15.0±2.4, 13.1±3.2, 13.3±3.1, 13.7±2.9, 13.2±2.4, and 13.4±2.3 mmHg (mean ± standard deviation), respectively. The correlation coefficient between tIOPCCT and tIOP (r=0.979) was significantly higher than that between tIOPCCT and the other IOPs (r=0.668–0.852; P<0.001, respectively). The correlation coefficient between IOPCCT and IOPg (r=0.994) or IOPcc and IOPg (r=0.892) was significantly higher than that between IOPCCT or IOPcc and the other IOPs (r=0.669–0.740; P<0.001, respectively). The correlation coefficient between GTCCT and GT (r=0.989) was significantly higher than that between GTCCT and the other IOPs (r=0.669–0.740; P<0.001, respectively).
Conclusion: The IOP adjusted by CCT or corneal biomechanical properties depends on the measurement instrument itself, rather than the adjustment methods, for eyes of patients with primary open-angle glaucoma.

Keywords: ocular response analyzer, corneal biomechanical property, corneal hysteresis, glaucoma, intraocular pressure

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Comparison of visual outcomes and subjective visual quality after bilateral implantation of a diffractive trifocal intraocular lens and blended implantation of apodized diffractive bifocal intraocular lenses

Kjell G Gundersen,¹ Rick Potvin<sup>2

1</sup>IFocus Øyeklinikk AS, Haugesund, Norway; ²Science in Vision, Akron, NY, USA

Purpose: To compare the visual acuity (VA) and quality of vision between bilateral implantation of a trifocal intraocular lens (IOL) and blended bifocal IOLs with an intermediate add in the dominant eye and a near add in the nondominant eye.
Patients and methods: Patients with either trifocal or blended bifocal IOLs implanted were recruited after surgery. Subjects returned for a single diagnostic visit between 3 and 24 months after surgery. VA was tested at various distances, including low-contrast acuity and acuity at their preferred reading distance. A binocular defocus curve was obtained, and subjective visual function and quality of vision were evaluated.
Results: Twenty-five trifocal subjects and 30 blended bifocal subjects were enrolled. There were no significant differences in low-contrast acuity, preferred reading distance, or acuity at that reading distance. Binocular vision at 4 m, 60 cm, and 40 cm was not statistically significantly different. The trifocal provided statistically significantly better visual acuity (P<0.05) at vergences from -0.5 to -1.5 D (from 2 m to 67 cm viewing distance, P<0.05). There was no statistically significant difference in the near vision subscale scores of the 39-question National Eye Institute Visual Function Questionnaire or the overall scores of the Quality of Vision questionnaire, though significantly more trifocal subjects reported that the observed visual disturbances were "bothersome" (P<0.05).
Conclusion: Both lens modalities provided subjects with excellent binocular near and distance vision, with similar low rates of visual disturbances and good reported functional vision. The trifocal IOL provided significantly better intermediate VA in the viewing distance range of 2 m to 67 cm, corresponding to viewing things such as a car dashboard or grocery shelf. VA was similar between groups at viewing distances from 60 to 40 cm, corresponding to computer or reading distance.

Keywords: ReSTOR, At LISA tri, multifocal IOL, cataract, presbyopia


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A randomized, double-blind, placebo-controlled study of oral antioxidant supplement therapy in patients with dry eye syndrome

Jehn-Yu Huang, Po-Ting Yeh, Yu-Chih Hou

Department of Ophthalmology, National Taiwan University Hospital, College of Medicine, National Taiwan University, Taipei, Taiwan

Purpose: To evaluate the efficacy of oral antioxidant supplementation in the treatment of patients with dry eye syndrome (DES).
Methods: A prospective, randomized, double-blinded study compared the effects of an antioxidant supplement (containing anthocyanosides, astaxanthin, vitamins A, C, and E, and several herbal extracts, including Cassiae semen and Ophiopogonis japonicus) with placebo on patients with DES. We assessed dry eye symptoms, visual acuity, Schirmer's test, tear film breakup time, cornea and conjunctiva fluorescein staining, serum anti-SSA/anti-SSB antibodies, and the level of reactive oxygen species (ROS) in tears. The supplementation period was 8 weeks and patients were followed up every 4 weeks for 16 weeks. A linear mixed model was used to compare the groups, while within-group differences were tested by repeated-measures analysis of variance.
Results: Forty-three patients, 20 and 23 in treatment and placebo groups, respectively, completed the study. Liver and renal functions were normal. Diastolic blood pressure decreased in the treatment group. There were no significant differences in systolic blood pressure, dry eye symptoms, serum anti-SSA and anti-SSB, visual acuity, intraocular pressure, or fluorescein corneal staining between the groups. Tear film breakup time scores and Schirmer's test without topical anesthesia significantly improved in the treatment group. Tear ROS level differed between the groups and decreased after treatment. Overall subjective impression revealed a significant improvement with treatment compared with placebo.
Conclusion: Oral antioxidant supplementations may increase tear production and improve tear film stability by reducing tear ROS. The vegetable-based antioxidant supplement used in this study is safe and can be utilized as an adjuvant therapy to conventional artificial tear therapy for patients with DES.

Keywords: dry eye, tear, reactive oxygen species, blood pressure, herbal extracts


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Effect of benzalkonium chloride-free latanoprost ophthalmic solution on ocular surface in patients with glaucoma

Tejaswini Walimbe,¹ Vidya Chelerkar,² Purvi Bhagat,³ Abhijeet Joshi,⁴ Atul Raut<sup>4

1</sup>Walimbe Eye Clinic, ²PBMA's H.V. Desai Eye Hospital, Pune, ³Glaucoma Clinic, M and J Western Regional Institute of Ophthalmology, Civil Hospital, Ahmedabad, ⁴Clinical Research Department, Sun Pharma Advanced Research Company Ltd., Mumbai, India

Introduction: Benzalkonium chloride (BAK), included as a preservative in many topical treatments for glaucoma, induces significant toxicity and alters tear breakup time (TBUT). BAK-containing latanoprost, an ester prodrug of prostaglandin F_2a, can cause ocular adverse events (AEs) associated with BAK. The purpose of this study was to evaluate the efficacy and safety of BAK-free latanoprost.
Patients and methods: A prospective, open-label, single-arm, multicenter, 8-week study in patients with primary open-angle glaucoma or ocular hypertension taking BAK-containing latanoprost for ≥12 months was performed. Patients were switched to BAK-free latanoprost ophthalmic solution 0.005% administered once daily, and eyes were assessed after 28 and 56 days. Primary efficacy and safety variables were TBUT and treatment-emergent AEs, respectively.
Results: At day 56, 40 eyes were evaluable. Mean TBUT increased significantly from baseline (3.67±1.60 seconds) to 5.03±2.64 and 6.06±3.39 seconds after 28 and 56 days of treatment with BAK-free latanoprost (P<0.0001). Ocular Surface Disease Index^© (OSDI^©) score also decreased significantly to 12.06±13.40 and 7.06±10.75 at 28 and 56 days, respectively, versus baseline (18.09±18.61, P<0.0001). In addition, inferior corneal staining score decreased significantly to 0.53 from baseline (0.85, P=0.0033). A reduction in conjunctival hyperemia and intraocular pressure was observed at both time points. No treatment-related serious AEs were evident and 12 (26.08%) treatment-emergent AEs occurred in seven patients, with eye pain and irritation being the most frequent. No clinically significant changes in vital signs or slit lamp examinations were observed.
Conclusion: Results indicate that switching from BAK-containing latanoprost to BAK-free latanoprost resulted in significant improvements in TBUT, OSDI^© score, and inferior corneal staining score, and measurable reductions in conjunctival hyperemia score. Furthermore, BAK-free latanoprost was well tolerated with only mild-to-moderate and self-limiting AEs. BAK-free latanoprost appears to be effective in protecting ocular surface integrity in glaucoma patients but further studies are needed to confirm this beneficial effect.

Keywords: tear breakup time, TBUT, Ocular Surface Disease Index, OSDI^©, inferior corneal staining score, conjunctival hyperemia, intraocular pressure, glaucoma, ocular surface


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A nonrandomized, open-label study to evaluate the effect of nasal stimulation on tear production in subjects with dry eye disease

Neil J Friedman,¹ Karla Butron,² Nora Robledo,² James Loudin,³ Stephanie N Baba,³ Arturo Chayet<sup>2

1</sup>Department of Ophthalmology, Stanford University, Mid-Peninsula Ophthalmology Medical Group, Palo Alto, CA, USA; ²Codet Vision Institute, Tijuana, Mexico; ³Oculeve, Inc., South San Francisco, CA, USA

Background: Dry eye disease (DED), a chronic disorder affecting the tear film and lacrimal functional unit, is a widely prevalent condition associated with significant burden and unmet treatment needs. Since specific neural circuits play an important role in maintaining ocular surface health, microelectrical stimulation of these pathways could present a promising new approach to treating DED. This study evaluated the efficacy and safety of nasal electrical stimulation in patients with DED.
Methods: This prospective, open-label, single-arm, nonrandomized pilot study included 40 patients with mild to severe DED. After undergoing two screening visits, enrolled subjects were provided with a nasal stimulation device and instructed to use it at home four times daily (or more often as needed). Follow-up assessments were conducted up to day 180. The primary efficacy endpoint was the difference between unstimulated and stimulated tear production quantified by Schirmer scores. Additional efficacy endpoints included change from baseline in corneal and conjunctival staining, symptoms evaluated on a Visual Analog Scale, and Ocular Surface Disease Index scores. Safety parameters included adverse event (AE) rates, visual acuity, intraocular pressure, slit-lamp biomicroscopy, indirect ophthalmoscopy, and endoscopic nasal examinations.
Results: Mean stimulated Schirmer scores were significantly higher than the unstimulated scores at all visits, and corneal and conjunctival staining and symptom scores from baseline to day 180 were significantly reduced. No serious device-related AEs and nine nonserious AEs (three device-related) were reported. Intraocular pressure remained stable and most subjects showed little or no change in visual acuity at days 30 and 180. No significant findings from other clinical examinations were noted.
Conclusion: Neurostimulation of the nasolacrimal pathway is a safe and effective means of increasing tear production and reducing symptoms of dry eye in patients with DED.

Keywords: keratoconjunctivitis sicca, neuromodulation, neurostimulation, nasolacrimal reflex, tear production, ocular staining


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Risk factors for visual impairment associated with corneal diseases in southern China

Sarah C Xu,¹ Jessica Chow,¹ Ji Liu,¹ Liang Li,² Jessica S Maslin,¹ Nisha Chadha,¹ Baihua Chen,² Christopher C Teng<sup>1

1</sup>Department of Ophthalmology and Visual Science, Yale University School of Medicine, New Haven, CT, USA; ²Department of Ophthalmology, The Second Xiangya Hospital of Central South University, Changsha, People's Republic of China

Purpose: To identify the most common etiologies of corneal disease and the risk factors associated with worse visual outcomes in Changsha, Hunan, located in southern China.
Methods: This observational, cross-sectional study evaluated 100 consecutive patients seen at the cornea clinic of The Second Xiangya Hospital of Central South University. Ocular history, demographic information, and ocular use of traditional Chinese medicine were recorded and analyzed. Causes of infectious keratitis were diagnosed clinically. Fungal and acanthamoeba keratitis were confirmed by confocal microscopy. Visual impairment was categorized based on visual acuity according to World Health Organization recommendations. A binary logistic regression model was used to calculate odds ratio (OR).
Results: One hundred consecutive patients were evaluated. Sixty patients (60%) had noninfectious corneal diseases, most commonly dry eye syndrome (26.7%, n=16), followed by corneal abrasion (18.3%, n=11). Forty-five patients had infectious keratitis, five of whom had both infectious and noninfectious etiologies. Of the patients with infectious keratitis, viral keratitis was the most frequent cause (57.8%, n=26), followed by fungal (20%, n=9) and bacterial (20%, n=9). Older age (OR =5.08, P=0.048), male sex (OR =3.37, P=0.035), and rural residence (OR =3.11, P=0.017) had increased odds of having worse visual impairment. Rural residence was also associated with infectious keratitis (P=0.005), particularly bacterial and fungal keratitis (P=0.046), and a history of ocular trauma (P=0.003). Occupation was not a significant risk factor in this population. Fourteen patients reported use of traditional Chinese medicine, with no association with visual outcomes found.
Conclusion: Older age, male sex, and rural residence were associated with worse visual impairment. Prevalence and outcome of corneal diseases may be improved with an increased awareness in these populations.

Keywords: rural area, international ophthalmology, corneal diseases, traditional Chinese medicine


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Pharmacokinetics and aqueous humor penetration of levofloxacin 1.5% and moxifloxacin 0.5% in patients undergoing cataract surgery

Frank A Bucci Jr, Ines Teuma Nguimfack, Angel T Fluet

Bucci Laser Vision Institute, Wilkes-Barre, PA, USA

Purpose: The objective of this study was to compare the pharmacokinetics of levofloxacin 1.5% and moxifloxacin hydrochloride 0.5% ophthalmic solutions in aqueous humor after multiple doses prior to cataract surgery.
Methods: Ninety-eight eyes underwent cataract surgery and met the requirements of PK analysis. Eligible eyes were randomly assigned in a 1:1 ratio to receive levofloxacin or moxifloxacin prior to cataract surgery and were randomized into one of four sampling time points (ie, 1, 2, 4, and 6 hour post-last dose). Randomization was investigator and laboratory-masked. Three days prior to cataract surgery, each patient instilled one drop of the assigned study medication into the operative eye four times daily. One aqueous humor specimen was collected from the eye at the randomized time point. Aqueous humor specimens were assayed for drug concentration using a validated liquid chromatography and tandem mass spectrometer.
Results: Concentrations of the drug in the aqueous humor, as described by mean C_max and pooled AUC_0–6 values, were greater for levofloxacin than moxifloxacin (C_max: 1.43, 0.87 µg/ml, respectively, P=0.008; AUC_0–6 6.1, 3.8 µg·min/ml, P<0.001 respectively). No treatment-emergent adverse events were reported.
Conclusion: Significantly greater drug exposures were attained in aqueous humor following the administration of levofloxacin 1.5% than moxifloxacin 0.5% ophthalmic solution. Achieving considerable higher drug concentration in the aqueous humor with levofloxacin 1.5% may demonstrate a greater potential for bacterial eradication.

Keywords: concentration, endophthalmitis, antibiotics, phacoemulsification, prophylaxis, fluoroquinolone


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Outcomes of excimer laser enhancements in pseudophakic patients with multifocal intraocular lens

Steven C Schallhorn,^1–3 Jan A Venter,² David Teenan,² Julie M Schallhorn,³ Keith A Hettinger,² Stephen J Hannan,² Martina Pelouskova<sup>2

1</sup>Department of Ophthalmology, University of California, San Francisco, CA, USA; ²Optical Express, Glasgow, UK; ³Department of Ophthalmology, University of Southern California, Los Angeles, CA, USA

Purpose: The aim of this study was to assess visual and refractive outcomes of laser vision correction (LVC) to correct residual refraction after multifocal intraocular lens (IOL) implantation.
Patients and methods: In this retrospective study, 782 eyes that underwent LVC to correct unintended ametropia after multifocal IOL implantation were evaluated. Of all multifocal lenses implanted during primary procedure, 98.7% were refractive and 1.3% had a diffractive design. All eyes were treated with VISX STAR S4 IR excimer laser using a convectional ablation profile. Refractive outcomes, visual acuities, patient satisfaction, and quality of life were evaluated at the last available visit.
Results: The mean time between enhancement and last visit was 6.3±4.4 months. Manifest spherical equivalent changed from -0.02±0.83 D (-3.38 D to +2.25 D) pre-enhancement to 0.00±0.34 D (-1.38 D to +1.25 D) post-enhancement. At the last follow-up, the percentage of eyes within 0.50 D and 1.00 D of emmetropia was 90.4% and 99.5%, respectively. Of all eyes, 74.9% achieved monocular uncorrected distance visual acuity 20/20 or better. The mean corrected distance visual acuity remained the same before (-0.04±0.06 logMAR [logarithm of the minimum angle of resolution]) and after LVC procedure (-0.04±0.07 logMAR; P=0.70). There was a slight improvement in visual phenomena (starburst, halo, glare, ghosting/double vision) following the enhancement. No sight-threatening complications related to LVC occurred in this study.
Conclusion: LVC in pseudophakic patients with multifocal IOL was safe, effective, and predictable in a large cohort of patients.

Keywords: excimer laser enhancements, pseudophakic patients, multifocal IOL

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Feasibility study of sustained-release travoprost punctum plug for intraocular pressure reduction in an Asian population

Shamira A Perera,^1–3 Daniel SW Ting,^1,2 Monisha E Nongpiur,^1–3 Paul T Chew,³ Maria Cecilia D Aquino,³ Chelvin CA Sng,³ Sue-Wei Ho,^1,2 Tin Aung<sup>1–3

1</sup>Singapore Eye Research Institute, Singapore; ²Singapore National Eye Centre, Singapore; ³Department of Ophthalmology, Yong Loo Lin School of Medicine, National University of Singapore, Singapore

Purpose: To investigate the efficacy and safety of a punctum plug-based sustained drug release system for a prostaglandin analog, travoprost (OTX-TP), for intraocular pressure (IOP) ­reduction in an Asian population.
Methods: This is an initial feasibility, prospective, single-arm study involving 26 eyes and a bioresorbable punctum plug containing OTX-TP. An OTX-TP was placed in the vertical portion of the superior or inferior canaliculus of patients with primary open-angle glaucoma or ocular hypertension. The main outcome measure was the IOP-lowering efficacy of OTX-TP at 3 (8 am) and 10, 20, and 30 days (8 am, 10 am, and 4 pm), compared to baseline.
Results: A total of 26 OTX-TP were inserted for 17 subjects. The mean (standard deviation) age was 57.2 (13.8) years. At 10 days, all plugs were still present, and the IOP reduction from baseline was 6.2 (23%), 5.4 (21%), and 7.5 mmHg (28%) at 8 am, 10 am, and 4 pm, respectively. At 10 days, the mean IOP (standard error of mean) was 21.2 (1.2), 20.4 (0.8), and 19.7 (1.0) at 8 am, 10 am, and 4 pm, respectively, showing no discernible IOP trend during the course of the day. At 30 days, plug retention had declined to 42%, and the overall IOP reduction had decreased to 16%.
Conclusion: The sustained-release OTX-TP is able to reduce IOP by 24% (day 10) and 15.6% (day 30), respectively. It is a potentially well-tolerable ocular hypotensive for glaucoma patients with a history of poor compliance.

Keywords: ocular hypertension, travoprost, bioresorbable, punctum plug, glaucoma, Asian


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Vitrectomy for center-involved diabetic macular edema

David J Browning,¹ Chong Lee,¹ Michael W Stewart,² Maurice B Landers III<sup>3

1</sup>Charlotte Eye, Ear, Nose and Throat Associates, Charlotte, NC, ²Mayo Clinic, Jacksonville, FL, ³Kittner Eye Center, University of North Carolina, Chapel Hill, NC, USA

Purpose: To determine the effect of vitrectomy for center-involved diabetic macular edema (CI-DME).
Methods: This was a retrospective study of 53 eyes of 45 patients who had vitrectomy for CI-DME and were followed up for at least 12 months. Charts were reviewed for visual acuity (VA), central subfield mean thickness measured by optical coherence tomography, presurgical and postsurgical interventions for CI-DME, and number of office visits in the first 12 months after surgery. Preoperative spectral domain optical coherence tomography was performed on 38 patients, and they were graded for ellipsoid zone (EZ) intactness by three independent graders with assessment of agreement between graders using intraclass correlation coefficients and Bland–Altman analysis.
Results: The median VA improved from 20/100 (interquartile range [IQR], 20/63–20/200) at baseline to 20/63 (IQR, 20/32–20/125) at 12 months. The median central subfield mean thickness improved from 505 µm (IQR, 389–597 µm) at baseline to 279 µm (IQR, 246–339 µm) at 12 months. Intergrader agreement for EZ intactness was moderate (intraclass correlation coefficients 0.4294–0.6356). There was no relationship between preoperative intactness of the EZ and the 12-month change in VA.
Conclusion: Vitrectomy consistently thins the macula in CI-DME and, on average, leads to clinically significant improvement in VA comparable in size to that reported with serial intravitreal anti-vascular endothelial growth factor injections. A large, comparative, prospective, randomized clinical trial of these two treatments is needed to determine which is more effective and cost-effective.

Keywords: center-involved diabetic macular edema, diabetic macular edema, vitrectomy, spectral domain OCT


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Glycan Node Analysis: A Bottom-up Approach to Glycomics

This article presents an enhanced form of a novel bottom-up glycomics technique designed to analyze the pooled compositional profile of glycans in unfractionated biofluids through the chemical breakdown of glycans into their constituent linkage-specific monosaccharides for detection by GC-MS. Potential applications include early detection of cancer and other glycan-affective disorders.

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Light-driven Enzymatic Decarboxylation

We describe a protocol for the light-catalyzed generation of hydrogen peroxide — a cofactor for oxidative transformations.

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Assessment of Startle Response and Its Prepulse Inhibition Using Posturography: Pilot Study

Purpose. The aim of this study was to evaluate the possibility of using static posturography in the assessment of sensorimotor gating. Subjects and Methods. Fourteen subjects took part in the experiment. The inhibitory mechanisms of startle reflex were used as the measure of sensorimotor gating. It was evoked by a strong acoustic stimulus (106 dB SPL, 40 ms) which was preceded by the weaker similar signal (80 dB SPL, 20 ms). A stabilographic platform was used to measure sensorimotor gating. Results. Results of static posturography show that the postural sway caused by the reaction to a strong acoustic stimulus is significantly smaller when this stimulus is preceded by the signal of lower intensity (prepulse). Such assessment is only possible in eyes open conditions. Conclusions. Static posturography can be simple and effective method used for diagnosis of sensorimotor gating in humans.

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Plasma miR-10a: A Potential Biomarker for Coronary Artery Disease

Aims. MicroRNAs (miRNAs) are involved in the pathogenesis of coronary artery disease (CAD). The objective of this study is to determine plasma levels of miR-10a in CAD and analyze its association with the severity of CAD. Materials and Methods. Plasma miR-10a levels in 60 CAD patients including stable angina pectoris (SAP) (), unstable angina pectoris (UAP) or non-ST elevation myocardial infarction (MI) (NSTEMI) (), or ST elevation MI (STEMI) () and 20 non-CAD subjects were assessed by real-time polymerase chain reaction (qRT-PCR), and associations of miR-10a levels with risk factors of CAD and its severity were analyzed. Results. The qRT-PCR results showed that plasma miR-10a levels were decreased in CAD patients, and CAD with high SYNTAX scores or STEMI was significantly associated with lower miR-10a levels. Conclusions. Lower plasma miR-10a levels were negatively associated with the presence as well as severity of CAD, and plasma miR-10a can act as a potential biomarker for estimating the presence and severity of CAD.

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Two-Year Follow-Up after Contact Force Sensing Radiofrequency Catheter and Second-Generation Cryoballoon Ablation for Paroxysmal Atrial Fibrillation: A Comparative Single Centre Study

Background. There are little comparative data on catheter ablation of paroxysmal atrial fibrillation (AF) using the contact force radiofrequency (CF-RF) catheter versus the second-generation cryoballoon (CB2). Methods and results. This is a single center, retrospective, nonrandomized study of 98 patients with symptomatic, drug-refractory paroxysmal AF who underwent their first PVI ablation using either the CB2 () or CF-RF (). The mean age was 60 years with 63% men, a mean LA size of 42 mm. The procedure duration ( versus minutes ) was shorter for CB2 group; the fluoroscopy time ( versus minutes, ) was similar. Complete PVI was achieved in 96% of patients with RF-CF and 98% with CB2. Phrenic nerve palsies (2 transient and 1 persistent) occurred exclusively in the CB2 group and 1 severe, nonlethal complication (pericardial tamponade) occurred in the CF-RF group. At 24-month follow-up, the success rate, defined as freedom from AF/atrial tachycardia (AT) after a single procedure without antiarrhythmic drug, was comparable in CF-RF group and CB2 group (65.5% versus 67%, resp., log rank ). Conclusion. Both the CB2 and the RF-CF ablation appeared safe; the success rate at 2 years was comparable between both technologies.

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The Utility of a Digital Virtual Template for Junior Surgeons in Pedicle Screw Placement in the Lumbar Spine

This study assessed the utility of three-dimensional preoperative image reconstruction as digital virtual templating for junior surgeons in placing a pedicle screw (PS) in the lumbar spine. Twenty-three patients of lumbar disease were operated on with bilateral PS fixation in our hospital. The two sides of lumbar pedicles were randomly divided into "hand-free group" (HFG) and "digital virtual template group" (DVTG) in each patient. Two junior surgeons preoperatively randomly divided into these two groups finished the placement of PSs. The accuracy of PS and the procedure time of PS insertion were recorded. The accuracy of PS in DVTG was 91.8% and that in HFG was 87.7%. The PS insertion procedure time of DVTG was s and that of HFG was s. Although no significant difference was reported in the accurate rate of PS between the two groups, the PS insertion procedure time was significantly shorter in DVTG than in HFG (). Digital virtual template is simple and can reduce the procedure time of PS placement.

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A Review of Animal Models of Intervertebral Disc Degeneration: Pathophysiology, Regeneration, and Translation to the Clinic

Lower back pain is the leading cause of disability worldwide. Discogenic pain secondary to intervertebral disc degeneration is a significant cause of low back pain. Disc degeneration is a complex multifactorial process. Animal models are essential to furthering understanding of the degenerative process and testing potential therapies. The adult human lumbar intervertebral disc is characterized by the loss of notochordal cells, relatively large size, essentially avascular nature, and exposure to biomechanical stresses influenced by bipedalism. Animal models are compared with regard to the above characteristics. Numerous methods of inducing disc degeneration are reported. Broadly these can be considered under the categories of spontaneous degeneration, mechanical and structural models. The purpose of such animal models is to further our understanding and, ultimately, improve treatment of disc degeneration. The role of animal models of disc degeneration in translational research leading to clinical trials of novel cellular therapies is explored.

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Volatiles from Subtropical Convolvulaceae That Interfere with Bacterial Cell-to-Cell Communication as Potential Antipathogenic Drugs

Increasing chronic bacterial infections create an urgent need for new antimicrobial agents or strategies for their control. Targeting virulence is one of the alternative approaches to find new medicines to treat persistent infections due to bacteria with biofilm-phenotype which are more resistant to antibiotics than their planktonic counterparts having an extreme capacity for evading the host defences. A bioguided study of sixteen extracts from flowers and leaves of four subtropical Convolvulaceae species provided evidence of the occurrence of antipathogenic natural products active against Gram positive and negative bacteria. Particularly, volatile metabolites from Merremia dissecta creeper, a food and medicinal plant, were able to interfere with the Pseudomonas aeruginosa quorum sensing system by a strong decrease of N-acyl homoserine lactone (AHL) biosynthesis (63–75%), which attenuated the virulence factor expression like biofilm (55%) and elastase activity (up to 27%), key factors that enable the colonization and dissemination of the infection in the host. Control of the P. aeruginosa biofilm and the QS process by phytochemicals, such as (+) spathulenol, isolated from a bioactive extract of M. dissecta leaves would be a good strategy for the development of new and effective antipathogenic drugs.

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Unveiling alterative splice diversity from human oligodendrocyte proteome data

Publication date: Available online 21 May 2016
Source:Journal of Proteomics
Author(s): Raphael Tavares, Gabriel Wajnberg, Nicole de Miranda Scherer, Bianca Alves Pauletti, Juliana S. Cassoli, Carlos Gil Ferreira, Adriana Franco Paes Leme, Patricia Savio de Araujo-Souza, Daniel Martins-de-Souza, Fabio Passetti
Oligodendrocytes produce and maintain the myelin sheath of axons in the central nervous system. Because misassembled myelin sheaths have been associated with brain disorders such as multiple sclerosis and schizophrenia, recent advances have been made towards the description of the oligodendrocyte proteome. The identification of splice variants represented in the proteome is as important as determining the level of oligodendrocyte-associated proteins. Here, we used an oligodendrocyte proteome dataset deposited in ProteomeXchange to search against a customized protein sequence file containing computationally predicted splice variants. Our approach resulted in the identification of 39 splice variants, including one variant from the GTPase KRAS gene and another from the human glutaminase gene family. We also detected the mRNA expression of five selected splice variants and demonstrated that a fraction of these have canonical proteins that may participate in direct protein-protein interactions. In conclusion, we believe our findings contribute to the molecular characterization of oligodendrocytes and may encourage other research groups working with central nervous system disorders to investigate the biological significance of these splice variants. The splice variants identified in this study may encode proteins that could be targeted in novel treatment strategies and diagnostic methods.SignificanceSeveral disorders of the central nervous system (CNS) are associated with misassembled myelin sheaths, which are produced and maintained by oligodendrocytes (OL). Recently, the OL proteome has been explored to identify key proteins and molecular functions associated with CNS disorders. We developed an innovative approach to select, with a higher level of confidence, a relevant list of splice variants from a proteome dataset and detected the mRNA expression of five selected variants: EEF1D, KRAS, MFF, SDR39U1, and SUGT1. We also described splice variants extracted from OL proteome data. Among the splice variants identified, some are from genes previously linked to CNS and related disorders. Our findings may contribute to oligodendrocyte characterization and encourage other research groups to investigate the biological role of splice variants and to improve current treatments and diagnostic methods for CNS disorders.Hightlights• Oligodendrocyte splice variants were investigated using a combined approach of proteogenomics and Mass Spectrometry• A customized protein repository identified 39 alternative splicing variants in the human oligodendrocyte proteome.• Five of those splice variants were confirmed at the transcriptome level in the MO3.13 cell line• Protein-protein network interactions were investigated



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Proteomic characterization of Nα- and Nε-acetylation in Acinetobacter baumannii

Publication date: Available online 21 May 2016
Source:Journal of Proteomics
Author(s): Takfarinas Kentache, Thierry Jouenne, Emmanuelle Dé, Julie Hardouin

Graphical abstract

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A proteomic signature of ovarian cancer tumor fluid identified by highthroughput and verified by targeted proteomics

Publication date: Available online 21 May 2016
Source:Journal of Proteomics
Author(s): Aline Poersch, Mariana Lopes Grassi, Vinícius Pereira de Carvalho, Guilherme Pauperio Lanfredi, Camila de Souza Palma, Lewis Joel Greene, Christiani Bisinoto de Sousa, Hélio Humberto Angotti Carrara, Francisco José Candido dos Reis, Vitor Marcel Faça
Tumor fluid samples have emerged as a rich source for the identification of ovarian cancer in the context of proteomics studies. To uncover differences among benign and malignant ovarian samples, we performed a quantitative proteomic study consisting of albumin immunodepletion, isotope labeling with acrylamide and in-depth proteomic profiling by LC-MS/MS in a pool of 10 samples of each histological type. 1135 proteins were identified, corresponding to 505 gene products. 223 proteins presented associated quantification and the comparative analysis of histological types revealed 75 differentially abundant proteins. Based on this, we developed a panel for targeted proteomic analysis using the multiple reaction monitoring (MRM) method for validation of 51 proteins in individual samples of high-grade serous ovarian tumor fluids (malignant) and benign serous cystadenoma tumor fluids. This analysis showed concordant results in terms of average amounts of proteins, and APOE, SERPINF2, SERPING1, ADAM17, CD44 and OVGP1 were statistically significant between benign and malignant group. The results observed in the MRM for APOE were confirmed by western blotting, where APOE was more abundant in malignant samples. This molecular signature can contribute to improve tumor stratification and shall be investigated in combination with current biomarkers in larger cohorts to improve ovarian cancer diagnosis.Biological significanceDespite advances in cancer research, ovarian cancer has a high mortality and remains a major challenge due to a number of particularities of the disease, especially late diagnosis caused by vague clinical symptoms, the cellular and molecular heterogeneity of tumors, and the lack of effective treatment. Thus, efforts are directed to better understand this neoplasia, its origin, development and, particularly the identification and validation of biomarkers for early detection of the disease in asymptomatic stage. In the present work, we confirmed by MRM method in individual ovarian tumor fluid samples the regulation of 27 proteins out of 33 identified in a highthroughput study. We speculate that the presence and/or differential abundance observed in tumor fluid is a cooperation primarily of high rates of secretion of such tumor proteins to extra tumor environment that will at the end accumulate in plasma, and also the accumulation of acute-phase proteins throughout the entire body. On top of that, consideration of physiological influences in the interpretation of expression observed, including age, menopause status, route-of-elimination kinetics and metabolism of the tumor marker, coexisting disease, hormonal imbalances, life-style influences (smoking, alcoholism, obesity), among others, are mandatory to enable the selection of good protein tumor marker candidates for extensive validation.

Graphical abstract

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Transcriptomic and proteomic analyses provide new insights into the regulation mechanism of low-temperature-induced leafy head formation in Chinese cabbage

Publication date: Available online 20 May 2016
Source:Journal of Proteomics
Author(s): Chang-wei Zhang, Yan-ping Wei, Dong Xiao, Li-wei Gao, Shan-wu Lyu, Xi-lin Hou, Guusje Bouuema
In Chinese cabbage, leafy head-related traits are directly related to the cabbage yield and marketability, which are often primarily concerned target for breeders. Although intensive studies has been on head formation in Chinese cabbage in the past decade, very scanty information is available on mechanism involved in the head formation under the influence of low temperature at transcriptome and proteme perspective. In this study, quantitative expression profiling based on RNASeq transcriptome and iTRAQ proteome were combined to investigate this trait for a global picture of the molecular dynamics. Total of 2931 transcripts and 365 proteins were identified with significantly changed level in abundance from heading and non-heading Chinese cabbage. Related analyses including function annotations, hierarchical categories, as well as the correlation from transcript-to-proteins were performed. The results indicated that the leafy head formation of Chinese cabbage has involved a complex regulatory pattern. The correlated genes that were mapped to the pathway of plant hormone signal transduction suggested that the head formation might be an integrated result of various plant hormones. Our combined analysis will provide a comprehensive approach to understanding the regulation mechanism of leafy head formation in Chinese cabbage.Biological Significance.This study revealed the direct relation of leafy-heading traits with the yield of the plant. A comprehensive investigation was done by integrating quantitative expression profiling analysis of transcriptome and proteomic to provide crucial information for further research on the molecular mechanism involved in head formation in Chinese cabbage. The correlation of transcript-to-protein in abundance may afford some necessary information of involvement of post-transcriptional factors influencing leafy head formation in Chinese cabbage.

Graphical abstract

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A cost-effective method to get insight into the peritoneal dialysate effluent proteome

Publication date: Available online 20 May 2016
Source:Journal of Proteomics
Author(s): J.E. Araújo, S. Jorge, F. Teixeira e Costa, A. Ramos, C. Lodeiro, H.M. Santos, J.L. Capelo

Graphical abstract

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Screening for target toxins of the antiophidic protein DM64 through a gel-based interactomics approach

Publication date: Available online 20 May 2016
Source:Journal of Proteomics
Author(s): Surza L.G. Rocha, Ana G.C. Neves-Ferreira, Monique R.O. Trugilho, Yamileth Angulo, Bruno Lomonte, Richard H. Valente, Gilberto B. Domont, Jonas Perales
DM64 is a glycosylated protein with antivenom activity isolated from the serum of the opossum Didelphis aurita. It binds non-covalently to myotoxins I (Asp49) and II (Lys49) from Bothrops asper venom and inhibits their myotoxic effect. In this study, an affinity column with immobilized DM64 as bait was used to fish potential target toxins. All ten isolated myotoxins tested were able to effectively bind to the DM64 column. To better access the specificity of the inhibitor, crude venoms from Bothrops (8 species), Crotalus (2 species) and Naja naja atra were submitted to the affinity purification. Venom fractions bound and nonbound to the DM64 column were analyzed by two-dimensional gel electrophoresis and MALDI-TOF/TOF MS. Although venom fractions bound to the column were mainly composed of basic PLA2, a few spots corresponding to acidic PLA2 were also observed. Some unexpected protein spots were also identified: C-type lectins and CRISP may represent putative new targets for DM64, whereas the presence of serine peptidases in the venom bound fraction is likely a consequence of nonspecific binding to the column matrix. The present results contribute to better delineate the inhibitory potential of DM64, providing a framework for the development of more specific antivenom therapies.Biological significanceLocal tissue damage induced by myotoxic PLA2 remains a serious consequence of snake envenomation, since it is only partially neutralized by traditional antivenom serotherapy. Myotoxin inhibition by highly specific molecules offers great promise in the treatment of snakebites, a health problem largely neglected by governments and pharmaceutical industries. Bioactive compounds such as DM64 can represent a valuable source of scaffolds for drug development in this area. The present study has systematically profiled the binding specificity of DM64 toward a variety of snake venom toxin classes and therefore can lead to a better understanding of the structure-function relationship of this important antivenom protein.

Graphical abstract

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Oncogenic KIT mutations in different exons lead to specific changes in melanocyte phospho-proteome

Publication date: Available online 20 May 2016
Source:Journal of Proteomics
Author(s): M. Sanlorenzo, I. Vujic, C. Posch, J. Ma, K. Lin, K. Lai, J.A. Oses-Prieto, S. Chand, J.L. Rodriguez-Peralto, A. Burlingame, S. Ortiz-Urda
Mutations in the proto-oncogene c-KIT (KIT) are found in several cancers, and the site of these mutations differs markedly between cancer types. We used site directed mutagenesis to induce KIT⁵⁵⁹, KIT⁶⁴² and KIT⁸¹⁶ mutations in primary human melanocytes (PHM) and we investigated the impact of each mutation on KIT function. We studied canonical KIT- signaling pathways by immunoblotting, and we used stable isotope labeling by amino acids in cell culture (SILAC) and kinase prediction models to identify kinases differently activated in respective mutants. We validated our results with the analysis of phosphorylation levels of selected substrates for each kinase. We concluded that CK1 ε and δ are more active in cell clones harboring KIT⁵⁵⁹ and KIT⁶⁴² mutations, whereas PAK4 is more active in clones with KIT⁸¹⁶ mutation. Our findings might help to develop further therapeutic options for tumors with specific KIT mutations in different domains.BIOLOGICAL SIGNIFICANCE.Different type of cancers harbor mutations in the oncogene KIT. The use of small molecules inhibitors directly targeting KIT had a limited success in the treatment of patients with KIT mutant cancers. Our study describes specific phospho-proteome changes due to different KIT mutations, and provides targets of further therapeutic options.

Graphical abstract

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From great kivas to great houses: Community formation and social dynamics in the southern Chuska Valley, New Mexico

Publication date: June 2016
Source:Journal of Anthropological Archaeology, Volume 42
Author(s): Monica L. Murrell, David T. Unruh
Understanding the sociopolitical contexts behind the construction of outlying Chacoan great houses requires a close examination of the relationships between great houses and their neighboring communities. Changes in household-ritual organization and socioeconomic interactions within the Figueredo great house community are used to examine the emergence of social complexity in the southern Chuska Valley. Long-term trends observed in settlement and economic organization are evaluated using a competition model to understand the social dynamics of local community development. Various lines of evidence suggest the residents of the Figueredo community comprised a multiethnic population that maintained a series of diverse socioeconomic interactions across the Four Corners region.



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Antigenic characterization of influenza viruses produced using synthetic DNA and novel backbones

Publication date: Available online 21 May 2016
Source:Vaccine
Author(s): Pirada Suphaphiphat, Lynne Whittaker, Ivna De Souza, Rodney S. Daniels, Philip R. Dormitzer, John W. McCauley, Ethan C. Settembre
The global system for manufacturing seasonal influenza vaccines has been developed to respond to the natural evolution of influenza viruses, but the problem of antigenic mismatch continues to be a challenge in certain years. In some years, mismatches arise naturally due to the antigenic drift of circulating viruses after vaccine strain selection has already been made. In other years, antigenic differences between the vaccine virus and circulating viruses are introduced as part of the current system, which relies on the use of egg-adapted isolates as a starting material for candidate vaccine viruses (CVVs). Improving the current process for making vaccine viruses can provide great value. We have previously established a synthetic approach for rapidly generating influenza viruses in a vaccine-approved Madin Darby canine kidney (MDCK) cell line using novel, high-growth backbones that increase virus rescue efficiency and antigen yield. This technology also has the potential to produce viruses that maintain antigenic similarity to the intended reference viruses, depending on the hemagglutinin (HA) and neuraminidase (NA) sequences used for gene synthesis. To demonstrate this utility, we generated a panel of synthetic viruses using HA and NA sequences from recent isolates and showed by hemagglutination inhibition (HI) tests that all synthetic viruses were antigenically-like their conventional egg- or cell-propagated reference strains and there was no impact of the novel backbones on antigenicity. This synthetic approach can be used for the efficient production of CVVs that may be more representative of circulating viruses and may be used for both egg- and cell-based vaccine manufacturing platforms. When combined with mammalian cell culture technology for antigen production, synthetic viruses generated using HA and NA sequences from a non-egg-adapted prototype can help to reduce the potential impact of antigenic differences between vaccine virus and circulating viruses on vaccine effectiveness.



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A national examination of pharmacy-based immunization statutes and their association with influenza vaccinations and preventive health

Publication date: Available online 21 May 2016
Source:Vaccine
Author(s): Kevin W. McConeghy, Coady Wing
BackgroundA series of state-level statute changes have allowed pharmacists to provide influenza vaccinations in community pharmacies. The study aim was to estimate the effects of pharmacy-based immunization statutes changes on per capita influenza vaccine prescriptions, adult vaccination rates, and the utilization of other preventive health services.MethodsA quasi-experimental study that compares vaccination outcomes over time before and after states allowed pharmacy-based immunization. Measures of per capita pharmacy prescriptions for influenza vaccines in each state came from a proprietary pharmacy prescription database. Data on adult vaccination rates and preventive health utilization were studied using multiple waves of the Behavioral Risk Factor Surveillance System (BRFSS). The primary outcomes were changes in per capita influenza vaccine pharmacy prescriptions, adult vaccination rates, and preventive health interventions following changes.ResultsBetween 2007 and 2013, the number of influenza vaccinations dispensed in community pharmacies increased from 3.2 to 20.9 million. After one year, adopting pharmacist immunization statutes increased per capita influenza vaccine prescriptions by an absolute difference (AD) of 2.6% (95% CI: 1.1–4.2). Adopting statutes did not lead to a significant absolute increase in adult vaccination rates (AD 0.9%, 95% CI: −0.3, 2.2). There also was no observed difference in adult vaccination rates among adults at high-risk of influenza complications (AD 0.8%, 95% CI: −0.2, 1.8) or among standard demographic subgroups. There also was no observed difference in the receipt of preventive health services, including routine physician office visits (AD −1.9%, 95% CI: −4.9, 1.1).ConclusionsPharmacists are providing millions of influenza vaccines as a consequence of immunization statutes, but we do not observe significant differences in adult influenza vaccination rates. The main gains from pharmacy-based immunization may be in providing a more convenient way to obtain an important health service.



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Report of the Cent Gardes HIV Vaccines Conference. Part 1: The antibody response; Fondation Mérieux Conference Center, Veyrier-du-Lac, France, 25–27 October 2015

Publication date: Available online 20 May 2016
Source:Vaccine
Author(s): Marc P. Girard, Roger Le-Grand, Valentina Picot, Christophe Longuet, Gary J. Nabel
The 2015 Cent Gardes Conference on HIV vaccines took place on October 25–27 at the Merieux Foundation Conference Center in Veyrier du Lac, near Annecy, France. The meeting reviewed progress in the development of HIV vaccines and identified new directions of future research. The field has advanced incrementally over the past year but major progress will require additional information from new clinical trials. In this article, we review the presentations on humoral immune responses to HIV, and highlight the difficulty of eliciting broadly neutralizing antibodies by vaccination. Advances in cellular immunity for HIV prevention will be reviewed separately, in a following article.



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The impact of implementing a demand forecasting system into a low-income country's supply chain

Publication date: Available online 21 May 2016
Source:Vaccine
Author(s): Leslie E. Mueller, Leila A. Haidari, Angela R. Wateska, Roslyn J. Phillips, Michelle M. Schmitz, Diana L. Connor, Bryan A. Norman, Shawn T. Brown, Joel S. Welling, Bruce Y. Lee
ObjectiveTo evaluate the potential impact and value of applications (e.g. ordering levels, storage capacity, transportation capacity, distribution frequency) of data from demand forecasting systems implemented in a lower-income country's vaccine supply chain with different levels of population change to urban areas.Materials and MethodsUsing our software, HERMES, we generated a detailed discrete event simulation model of Niger's entire vaccine supply chain, including every refrigerator, freezer, transport, personnel, vaccine, cost, and location. We represented the introduction of a demand forecasting system to adjust vaccine ordering that could be implemented with increasing delivery frequencies and/or additions of cold chain equipment (storage and/or transportation) across the supply chain during varying degrees of population movement.ResultsImplementing demand forecasting system with increased storage and transport frequency increased the number of successfully administered vaccine doses and lowered the logistics cost per dose up to 34%. Implementing demand forecasting system without storage/transport increases actually decreased vaccine availability in certain circumstances.DiscussionThe potential maximum gains of a demand forecasting system may only be realized if the system is implemented to both augment the supply chain cold storage and transportation. Implementation may have some impact but, in certain circumstances, may hurt delivery. Therefore, implementation of demand forecasting systems with additional storage and transport may be the better approach. Significant decreases in the logistics cost per dose with more administered vaccines support investment in these forecasting systems.ConclusionDemand forecasting systems have the potential to greatly improve vaccine demand fulfilment, and decrease logistics cost/dose when implemented with storage and transportation increases direct vaccines. Simulation modeling can demonstrate the potential health and economic benefits of supply chain improvements.



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Humoral, T-cell and B-cell immune responses to seasonal influenza vaccine in solid organ transplant recipients receiving anti-T cell therapies

Publication date: Available online 21 May 2016
Source:Vaccine
Author(s): Delphine Héquet, Manuel Pascual, Sarah Lartey, Rishi D. Pathirana, Geir Bredholt, Katja Hoschler, Roger Hullin, Pascal Meylan, Rebecca J. Cox, Oriol Manuel
BackgroundWe analyzed the impact of the anti-T-cell agents basiliximab and antithymocyte globulins (ATG) on antibody and cell-mediated immune responses after influenza vaccination in solid-organ transplant recipients.Methods71 kidney and heart transplant recipients (basiliximab [n=43] and ATG [n=28]) received the trivalent influenza vaccine. Antibody responses were measured at baseline and 6 weeks post-vaccination by hemagglutination inhibition assay; T-cell responses were measured by IFN-γ ELISpot assays and intracellular cytokine staining (ICS); and influenza-specific memory B-cell (MBC) responses were evaluated using ELISpot.ResultsMedian time of vaccination from transplantation was 29 months (IQR 8–73). Post-vaccination seroconversion rates were 26.8% for H1N1, 34.1% for H3N2 and 4.9% for influenza B in the basiliximab group and 35.7% for H1N1, 42.9% for H3N2 and 14.3% for influenza B in the ATG group (p=0.44, p=0.61, and p=0.21, respectively). The number of influenza-specific IFN-γ-producing cells increased significantly after vaccination (from 35 to 67.5 SFC/10⁶ PBMC, p=0.0007), but no differences between treatment groups were observed (p=0.88). Median number of IgG-MBC did not increase after vaccination (H1N1, p=0.94; H3N2 p=0.34; B, p=0.79), irrespective of the type of anti-T-cell therapy.ConclusionsAfter influenza vaccination, a significant increase in antibody and T-cell immune responses but not in MBC responses was observed in transplant recipients. Immune responses were not significantly different between groups that received basiliximab or ATG.



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Effectiveness of seasonal trivalent influenza vaccination against hospital-attended acute respiratory infections in pregnant women: A retrospective cohort study

Publication date: Available online 20 May 2016
Source:Vaccine
Author(s): Annette K. Regan, Nicholas de Klerk, Hannah C. Moore, Saad B. Omer, Geoffrey Shellam, Paul V. Effler
BackgroundPregnant women are at risk of serious influenza infection. Although previous studies indicate maternal influenza vaccination can prevent hospitalisation in young infants, there is limited evidence of the effect in mothers.MethodsA cohort of 34,701 pregnant women delivering between 1 April 2012 and 31 December 2013 was created using birth records. Principal diagnosis codes from hospital emergency department (ED) and inpatient records were used to identify episodes of acute respiratory illness (ARI) during the 2012 and 2013 southern hemisphere influenza seasons. Cox regression models were used to calculate adjusted hazard ratios (aHRs) by maternal vaccination status, controlling for Indigenous status, socioeconomic level, medical conditions, and week of delivery.Results3,007 (8.7%) women received a seasonal influenza vaccine during pregnancy. Vaccinated women were less likely to visit an ED during pregnancy for an ARI (9.7 visits per 10,000 person-days vs. 35.5 visits per 10,000 person-days; aHR: 0.19, 95% CI: 0.05−0.68). Vaccinated women were also less likely to be hospitalised with an ARI compared to unvaccinated women (16.2 hospitalisations per 10,000 person-days vs. 34.0 hospitalisations per 10,000 person-days; aHR: 0.35, 95% CI: 0.13−0.97).ConclusionsInfluenza vaccination during pregnancy was associated with significantly fewer hospital attendances for ARI in pregnant women.



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Ultrastructural Changes and Death of Leishmania infantum Promastigotes Induced by Morinda citrifolia Linn. Fruit (Noni) Juice Treatment

The search for new treatments against leishmaniasis has increased due to high frequency of drug resistance registered in endemics areas, side effects, and complications caused by coinfection with HIV. Morinda citrifolia Linn., commonly known as Noni, has a rich chemical composition and various therapeutic effects have been described in the literature. Studies have shown the leishmanicidal activity of M. citrifolia; however, its action on the parasite has not yet been elucidated. In this work, we analyzed leishmanicidal activity and ultrastructural changes in Leishmania infantum promastigotes caused by M. citrifolia fruit juice treatment. M. citrifolia fruit extract showed a yield of 6.31% and high performance liquid chromatography identified phenolic and aromatic compounds as the major constituents. IC50 values were 260.5 µg/mL for promastigotes and 201.3 µg/mL for intracellular amastigotes of L. infantum treated with M. citrifolia. Cytotoxicity assay with J774.G8 macrophages showed that M. citrifolia fruit juice was not toxic up to 2 mg/mL. Transmission electron microscopy showed cytoplasmic vacuolization, lipid inclusion, increased exocytosis activity, and autophagosome-like vesicles in L. infantum promastigotes treated with M. citrifolia fruit juice. M. citrifolia fruit juice was active against L. infantum in the in vitro model used here causing ultrastructural changes and has a future potential for treatment against leishmaniasis.

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Responding to the existentials of non-life-threatening chronic conditions

Publication date: August 2016
Source:Medical Hypotheses, Volume 93
Author(s): Peter J. Adams
Diagnosis of a major life-threatening illness can prompt a range of existential responses that typically include strong emotions and a reassessment of priorities. What has not attracted attention are the existential responses to being told about less major but permanent medical conditions. A conceptual frame is developed for identifying and speaking about the existential dynamics of permanent change. It introduces two key concepts, finitude and transcendence, and uses them to explain the common sequence of responses first from being told about a chronic condition through to when its presence has become integrated into everyday life. Based on this analysis, a range of recommendations are provided on what types of conversations are helpful through the various stages of adjustment.



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Home-applied IPL epilation may prevent the problems due to hair follicle in amputees

Publication date: August 2016
Source:Medical Hypotheses, Volume 93
Author(s): S. Ergözen, E. Ercan, Y. Demir
Amputees may suffer from dermatoses such as folliculitis and pilonidal sinus caused by pressure on the stump in hairy parts of the skin. These conditions commonly cause pain and need treatment that result in abandonment of prosthesis use and disrupt the patient's daily living activities. We believe these conditions may be easily, effectively and cheaply prevented with the use of intense pulsed light technology, which is produced for home epilation. The use of this kind of epilator in the early period post-amputation may prevent the development of folliculitis and pilonidal sinus. This application may also be cost-effective in long term.



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General vulnerability to stuttering: The experience of stuttering and conditions bringing about invulnerability

Publication date: August 2016
Source:Medical Hypotheses, Volume 93
Author(s): P.M. Briley, J.S. Kalinowski
General vulnerability to stuttering is the broad awareness of stuttering and the ever-present, experiential sense of a person who stutters (PWS). It is defined by stuttering in all its forms and the awareness of its presence, both in moments of stuttering and moments of perceivably fluent speech. Under the heading of general vulnerability to stuttering is specific vulnerability to stuttering, which includes the actual events of stuttering (i.e., overt symptoms, covert symptoms, subperceptual stuttering, and anticipation of stuttering). The differentiation between the two is that specific vulnerability requires a specific moment of stuttering where general vulnerability does not.



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