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Additional file of Validation and standardization of DNA extraction and library construction methods for metagenomics-based human fecal microbiome measurements

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Arjun Shrestha,1 Bijay Khatri,2 Takeshi Naito3 1Department of Ophthalmology, BP Eye Foundation, Hospital for Children, Eye, ENT & Rehabilitation Services, Bhaktapur, Bagmati Province, Nepal; 2Academic and Research Department, BP Eye Foundation, Hospital for Children, Eye, ENT & Rehabilitation Services, Bhaktapur, Bagmati Province, Nepal; 3Department of Ophthalmology, Tokushima University Hospital, Tokushima, JapanCorrespondence: Arjun ShresthaDepartment of Ophthalmology, BP Eye Foundation, Hospital for Children, Eye, ENT & Rehabilitation Services, Bhaktapur, Bagmati Province, NepalTel +977 1 6631705Fax + 977 1 6639735Email drarjuns@hotmail.comBackground: Recently, the number of retinal patients is increasing in Nepal. The retinal problems are the second common cause of blindness in Nepal. Retinal diseases are challenging to manage due to the lack of simple and cost-effective screening methods, limited human resources, poverty, and lack of awareness in Nepal. Hospital for Children, Eye, ENT & Rehabilitation Services (CHEERS), Nepal conducted retina screening camps and education in communities through Retina Eye Care of Nepal project (RECON) in the years 2017 and 2018.Materials and Methods: We screened retinal patients with an indirect ophthalmoscope coupled with plus 20 Dioptre lens and portable fundus camera. We referred the patients needing interventions to the base hospital (CHEERS) using a screening algorithm, where they received retinal laser and surgery. Besides, we also provided on-site educational programs for retinal disease awareness to the community.Results: Retinal problems found from two static outreach clinics (SORC) and a day screening and treatment service (DSTS) were 18.3%, 20%, and 20%, respectively, while from a remote rural health camp (RRHC) was 2.61%. Likewise, patients referred to CHEERS for retinal laser or other retinal interventions were 15%, 16.6%, 16%, and 1.96% from 2 SORCs, 1 DSTS, and 1 RRHC, respectively.Conclusion: Retina camp is a cost-effective approach to early detection and referral for retinal-related vision impairment patients. It is also one opportunity to educate the community. The purpose of the project was to provide retina care to the community who otherwise could not afford it. We recommend a similar screening model on a large scale in a low-resource setting for the maximum benefits to the rural community.Keywords: screening, retina camp, Nepal, outreach

We consider some restricted games which are generalization of the games with graph-communication structure. We propose a result of in-essential coalitions structure of some intersecting convex games. Also we show a relationship between link-convexity and d-revised link-convexity for games on some intersecting family.

Purpose : To compare visual outcomes, intraoperative and postoperative complications of 25-gauge transconjunctival sutureless vitrectomy (25-G TSV) versus conventional 20-gauge vitrectomy in epiretinal membrane (ERM) surgery. Methods : A retrospective comparative study was conducted on 200 eyes with idiopathic ERM who underwent either primary 20-gauge vitrectomy (n=95) or 25-G TSV (n=105) with 6 months follow up. The following parameters were collected and compared : age, gender, best corrected visual acuity (BCVA), intraocular pressure (IOP), postoperative anterior chamber inflammation, intraoperative and postoperative complications. Results : Twenty-five gauge group achieved a significantly better vision at the 1st postoperative month (p=0.008) and a significantly higher incidence of visual improvement 6 months postoperatively (p=0.04) than 20-gauge group. Intraocular pressure was significantly higher at the 1st, 3rd and 6th postoperative months compared with baseline in the 20-G group. However, in the 25-gauge group IOP remained near to the baseline at the same period. Twenty five-gauge group achieved a significantly lower anterior chamber inflammation and a lower incidence of postoperative cystoid macular edema (CME) (p=0.01), hyphema (p=0.000) and after cataract (p=0.000) than 20-gauge group. Conclusions : The 25-gauge sutureless vitrectomy achieves rapid and higher visual improvement, less postoperative inflammation and complications than the 20-gauge vitrectomy in ERM surgery.

Purpose : To compare the incidence of wound sealing-related complications of 25-gauge transconjunctival sutureless vitrectomy (25-G TSV) for proliferative diabetic retinopathy (PDR) versus non-diabetic simple macular pathology (SMP). Methods : A retrospective comparative study was conducted on 377 eyes that underwent 25-G TSV for PDR (n=189 eyes) or non-diabetic SMP (n=188 eyes). Both groups were compared retrospectively and followed for at least 1 month (range 1-6 months). Main outcome measure was the incidence of postoperative wound sealing-related complications in both groups. Results : We found a statistically significant lower incidence of wound sealing-related complications on the 1st postoperative day in the PDR than SMP group (P-value 0.05). The incidence were as follows : 9% vs 18% for hypotony, 0% vs 2% for choroidal detachment, 0% vs 2% for external leakage and 1% vs 8% for conjunctival bleb formation, for the PDR and SMP groups respectively. This was associated with a significantly higher postoperative inflammation and IOP in PDR group. The day 1 higher postoperative IOP and inflammation were correlated with the presence of PDR pathology. Conclusion : The higher postoperative inflammation and IOP following vitrectomy in diabetic eyes, especially when combined with endolaser photocoagulation, makes such eyes less susceptible to wound sealing-related complications following sutureless vitrectomy.

We present a case of prepapillary vascular loops complicated by a suspected macroaneurysm rupture which was treated with intravitreal bevacizumab (IVB). A 62-year-old woman presented with decreased vision and myodesopsia in her left eye. Her best-corrected visual acuity (BCVA) was 0.6 in the left eye. Fundus examination disclosed an elevated, round, and reddish lesion, retinal hemorrhage at the superior aspect of the optic disc, retinal opacification along the superior branch retinal artery, and a small vitreous hemorrhage. Optical coherence tomography showed a serous retinal detachment, and indocyanine green angiography demonstrated prepapillary vascular loops and a hypofluorescent area with hyperfluorescent margins. These findings suggested the presence of a macroaneurysm. No filling of the dye in the aneurysm-like dilatation suggested a blockage of the lumen with a thrombus which might be associated with a branch retinal artery occlusion (BRAO). A diagnosis of prepapillary vascular loops complicated by a suspected macroaneurysm rupture and BRAO was made. Because of a persistent serous retinal detachment, IVB was performed. One month later, the BCVA improved to 1.0. Fundus examination disclosed an organized yellowish-white macroaneurysm and resolution of the serous retinal detachment. We recommend careful monitoring of patients with prepapillary vascular loops because of complications such as macroaneurysm rupture and BRAO.

With recent development of spectral-domain optical coherence tomography (SD-OCT), the pathological changes of retina can be observed in much greater detail. SD-OCT clearly delineates three highly reflective lines in the outer retina, which are external limiting membrane (ELM), photoreceptor inner and outer segment (IS/OS) junction, and cone outer segment tips (COST) in order from inside. These lines can serve as hallmarks for the evaluation of photoreceptor condition. In retinitis pigmentosa (RP) leading to photoreceptor degeneration, the ELM, IS/OS, and COST lines are shortened with the progression of the disease. In addition, shortening of the ELM, IS/OS and COST lines is significantly associated with each other. The line length is longest in the ELM, followed by the IS/OS, and COST, suggesting that retinal layer becomes disorganized first at the COST, followed by the IS/OS and finally the ELM. This finding is consistent with the previous report that the earliest histopathological change in RP is a shortening of the photoreceptor outer segments. On the other hand, retinal layer becomes restored first at the ELM, followed by the IS/OS and finally the COST after macular hole surgery. There may be a directionality of photoreceptor impairment or restoration on optical coherence tomographic image.