Noninvasive Detection of Mitochondrial Dysfunction in Ocular Hypertension and Primary Open-angle Glaucoma.

PURPOSE: To assess mitochondrial dysfunction in vivo in ocular hypertension (OHT) and primary open-angle glaucoma (POAG) using retinal metabolic analysis. PATIENTS AND METHODS: This was an observational, cross-sectional study performed from November 2015 to October 2016 at the New York Eye and Ear Infirmary of Mount Sinai. Thirty-eight eyes with varying stages of POAG, 16 eyes with OHT, and 32 control eyes were imaged on a custom fundus camera modified to measure full retinal thickness fluorescence at a wavelength optimized to detect flavoprotein fluorescence (FPF). Optical coherence tomography was used to measure the retinal ganglion cell-plus layer (RGC+) thickness. Macular FPF and the ratio of macular FPF to RGC+ thickness were the primary outcome variables and were compared among the three groups using an age-adjusted linear regression model. A mixed-effects model was used to assess correlations between FPF variables and clinical characteristics. RESULTS: Both macular FPF and the macular FPF/RGC+ thickness ratio were significantly increased in OHT compared with control eyes (P<0.05 and <0.01, respectively). In POAG eyes, macular FPF was not significantly increased compared with controls (P=0.24). However, the macular FPF/RGC+ thickness ratio in POAG eyes was significantly increased compared with controls (P<0.001). FPF was significantly correlated to age in POAG eyes. CONCLUSIONS: Despite lacking clinical evidence of glaucomatous deterioration, OHT eyes displayed significantly elevated macular FPF, suggesting that mitochondrial dysfunction may be detected before structural changes visible on current clinical imaging. Our preliminary results suggest that macular FPF analysis may prove to be a useful tool in assessing and evaluating OHT and POAG eyes.

Peripapillary perfused capillary density in primary open-angle glaucoma across disease stage: an optical coherence tomography angiography study.

AIMS: To assess peripapillary perfused capillary density (PCD) in primary open-angle glaucoma (POAG) across stage of disease. METHODS: In this observational, cross-sectional study, 60 eyes with varying stages of POAG and 24 control eyes were imaged on a spectral domain optical coherence tomography angiography system (AngioVue, Optovue, Fremont, California, USA) generating images centred on the optic nerve head. Major blood vessels were removed using custom automated software. PCD was calculated as a percentage as the ratio of pixels associated with perfused capillaries to the total number of pixels in the corresponding region-of-interest (ROI). Analysis of covariance was used to compare PCD among the subject groups and control for possible covariates. Area under the receiver operating characteristic curve (AROC) and sensitivity at 95% specificity were calculated to assess the capability of PCD to distinguish mild glaucoma from control. The Pearson's product-moment correlation coefficient was used to assess correlations between PCD and circumpapillary retinal nerve fibre layer thickness (cpRNFLT) and visual field mean deviation (MD). RESULTS: PCD demonstrated a progressive stepwise decrease from control eyes throughout worsening POAG stage at all ROIs. PCD demonstrated AROC and sensitivity values comparable to cpRNFLT and visual field parameters and exhibited significant correlations with cpRNFLT and MD at all corresponding ROIs. CONCLUSIONS: PCD displayed significant correlations with morphological and functional indices and exhibited diagnostic capabilities comparable to currently employed clinical variables. Our preliminary results suggest that PCD analysis may prove to be a useful tool in monitoring POAG across stage and identifying early POAG.

Visualization of Radial Peripapillary Capillaries Using Optical Coherence Tomography Angiography: The Effect of Image Averaging.

OBJECTIVES: To assess the effect of image registration and averaging on the visualization and quantification of the radial peripapillary capillary (RPC) network on optical coherence tomography angiography (OCTA). METHODS: Twenty-two healthy controls were imaged with a commercial OCTA system (AngioVue, Optovue, Inc.). Ten 10x10° scans of the optic disc were obtained, and the most superficial layer (50-µm slab extending from the inner limiting membrane) was extracted for analysis. Rigid registration was achieved using ImageJ, and averaging of each 2 to 10 frames was performed in five ~2x2° regions of interest (ROI) located 1° from the optic disc margin. The ROI were automatically skeletonized. Signal-to-noise ratio (SNR), number of endpoints and mean capillary length from the skeleton, capillary density, and mean intercapillary distance (ICD) were measured for the reference and each averaged ROI. Repeated measures analysis of variance was used to assess statistical significance. Three patients with primary open angle glaucoma were also imaged to compare RPC density to controls. RESULTS: Qualitatively, vessels appeared smoother and closer to histologic descriptions with increasing number of averaged frames. Quantitatively, number of endpoints decreased by 51%, and SNR, mean capillary length, capillary density, and ICD increased by 44%, 91%, 11%, and 4.5% from single frame to 10-frame averaged, respectively. The 10-frame averaged images from the glaucomatous eyes revealed decreased density correlating to visual field defects and retinal nerve fiber layer thinning. CONCLUSIONS: OCTA image registration and averaging is a viable and accessible method to enhance the visualization of RPCs, with significant improvements in image quality and RPC quantitative parameters. With this technique, we will be able to non-invasively and reliably study RPC involvement in diseases such as glaucoma.

Laparoscopic Tubal Reanastomosis Using Barbed Sutures.

STUDY OBJECTIVE: To show a new technique of laparoscopic tubal reanastomosis using barbed sutures. DESIGN: Step-by-step explanation of the technique using videos. SETTING: Laparoscopic tubal reanastomosis is an effective procedure with a high success rate for reversal of tubal sterilization. Conventionally, 4 equidistant interrupted sutures are placed under a magnified view for laparoscopic tubal reanastomosis. This step demands high precision and requires a lot of skill and experience. We have tried to simplify this suturing technique by using barbed sutures because they do not require knotting. Two separate 5-0 Quill barbed sutures (Angiotech Puerto Rico Inc, Aguadilla, Puerto Rico) are used in this technique. The first suture is used for taking 6 and 3 o' clock stitches. The second suture is used for taking 9 and 12 o' clock stitches. With this technique, the purse-string effect on the tubal lumen is reduced. INTERVENTIONS: Laparoscopic tubal reanastomosis using 5-0 Quill barbed sutures (equivalent to United States Pharmacopeia suture size 6-0). CONCLUSION: This technique of laparoscopic tubal reanastomosis using barbed sutures is a feasible and simpler alternative to conventional suturing.

Technology and the Glaucoma Suspect.

PURPOSE: To determine and compare the diagnostic performance of spectral-domain optical coherence tomography (SD-OCT), stereoscopic disc photographs, and automated perimetry as assessed by a group of glaucoma specialists in differentiating individuals with early glaucoma from suspects. METHODS: Forty-six eyes (46 patients) with suspicious optic nerves had previously undergone SD-OCT scans, 24-2 visual fields (VFs), and optic disc photographs. The average VF mean deviation was -1.97 ± 2.09 (SD) dB. Four glaucoma specialists examined the 138 individual diagnostic tests and classified the patient as likely glaucomatous or nonglaucomatous based on the results of a single test. The diagnostic performances of each of the three tests were compared to a previously determined reference standard, based on the consensus of a separate panel of four glaucoma specialists who examined all three tests together. RESULTS: Among the four specialists, the interobserver agreement across the three diagnostic tests was poor for VF and photos, with kappa (κ) values of 0.13 and 0.16, respectively, and moderate for OCT, with κ value of 0.40. Using panel consensus as reference standard, OCT had the highest discriminative ability, with an area under the curve (AUC) of 0.99 (95% 0.96-1.0) compared to photograph AUC 0.85 (95% 0.73-0.96) and VF AUC 0.86 (95% 0.76-0.96), suggestive of closer performance to that of a group of glaucoma specialists. CONCLUSIONS: Compared to VF and disc photography, SD-OCT, when used alone, had better internal agreement as well as better agreement with the consensus of clinicians using all available data. Future studies should evaluate best practices for SD-OCT interpretation.

Contained Morcellation for Laparoscopic Myomectomy Within a Specially Designed Bag.

A technique of contained morcellation of uterine myomas within a bag specially designed for 2-port morcellation during laparoscopic myomectomy is described. Ten patients underwent in-bag morcellation of myomas with a tissue isolation bag (MorSafe) between November 2014 and January 2015. The MorSafe tissue isolation bag is a retort-shaped bag made of medical-grade flexible plastic material with the wider opening of 134 mm in diameter and the tail end measuring 4 mm in diameter, allowing easy accomodation of specimens up to 12 cm in diameter. This technique involves placing the myomas into the isolation bag within the abdomen, exteriorizing the tail end of the bag, insufflating the bag within the peritoneal cavity, and morcellating the myomas under vision. Demographic and perioperative characteristics were studied. The mean operative time was 117 minutes (range, 75-195 minutes), the mean time for specimen introduction into the bag was 12.5 minutes (range, 7-22 minutes), and the mean time for morcellation and bag removal was 24.8 minutes (range, 10-50 minutes). There were no complications related to the in-bag morcellation technique, and there was no visual evidence of damage to the isolation bag. In-bag morcellation using this new bag is a feasible technique for morcellating uterine myomas in a contained manner and may provide an option to minimize the risks of open power morcellation while preserving the benefits of minimally invasive surgery.

Uterine Sarcomas in Patients Undergoing Surgery for Presumed Leiomyomas: 10 Years' Experience.

STUDY OBJECTIVE: To find the incidence of sarcomas in patients undergoing surgery for presumed leiomyomas. DESIGN: Retrospective study (Canadian Task Force classification II-3). SETTING: Paul's Hospital, Centre for Advanced Laparoscopy and Infertility, Kochi, India. PATIENTS: All women who underwent total laparoscopic hysterectomy (TLH), laparoscopic myomectomy (LM), and hysteroscopic myomectomy (HM) at Paul's Hospital for presumed leiomyomas from January 1, 2004 to December 31, 2014 that were then diagnosed as sarcomas on histopathologic examination. INTERVENTIONS: TLH, LM, and HM. MEASUREMENTS AND MAIN RESULTS: A total of 2678 patients underwent TLH, LM, and HM at Paul's Hospital for presumed leiomyomas from January 1, 2004 to December 31, 2014. Five patients were diagnosed as leiomyosarcoma and 3 patients as endometrial stromal sarcoma on histopathologic examination. Women's ages ranged from 12 to 53 years. Histopathologic diagnosis of leiomyosarcoma was made in 3 patients from the TLH group and 2 patients from the myomectomy group. Two patients from the TLH group and 1 patient from the myomectomy group were diagnosed as endometrial stromal sarcoma on histopathologic examination. The incidence of uterine sarcomas (leiomyosarcoma and endometrial stromal sarcoma) in patients undergoing surgery for presumed leiomyomas was found to be .29% (1 in 335 patients) in our study. CONCLUSION: Over a period of 10 years (2004-2014), the incidence of uterine sarcomas in patients undergoing surgery for presumed leiomyomas was found to be .29% in our study.

Deprotonation of 1,2-dialkylpyridinium ions: a DFT study of reactivity and site selectivity.

A site-selectivity model, based on the Fukui function as a local reactivity descriptor, has been applied to 1,2-disubstituted pyridinium ions incorporating two competing sites of similar reactivity, i.e., 1-methylene and 2-methylene, which may undergo deprotonation depending on the nature of substituent present on these moieties. Applicability of the local HSAB rule, in context with the Li-Evans' generalized HSAB principle suggesting the hard-hard interactions to be controlled by minimum Fukui function, has been illustrated. Global and local reactivity descriptors have been computed by carrying out DFT calculations at B3LYP/6-31++G** level using Mulliken and NPA methods for charge analysis. A comparison with the calculated deprotonation energies involving two sites indicates that the observed site selectivity in differently substituted pyridinium ions is better explained by the Li-Evans rule of minimum Fukui function for hard-hard interactions.

Substituent-dependent anomeric effects as a source of conformational preference in pyridinium methylides.

A systematic density functional theory level investigation of differently substituted pyridinium methylides was carried out to determine the role of C(ylidic) lone-pair-associated hyperconjugative and negative hyperconjugative interactions in deciding conformational preferences. Deviation from the coplanar orientation of the carbanionic center with the pyridine ring and its substituent dependence has been found to correlate well with the relative opportunities for conjugative and negative hyperconjugative interactions of a ylidic moiety with different substituent groups present at the ylidic carbon. The contribution of individual n-->pi* conjugative, n-->sigma* negative hyperconjugative, and sigma-->pi* hyperconjugative interactions in a particular conformation of pyridinium dichlorophosphinomethylides was assessed from donor-acceptor stabilization energies, as obtained from natural bond orbital (NBO) analysis. The relative extent of conjugative and negative hyperconjugative interactions with the substituents present at the ylidic carbon plays an important role in permitting the delocalization of ylidic charge into the pyridine ring, thereby controlling the relative orientation of the latter with the carbanionic plane.