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Plants in either raw form or their root extracts are utilized as complementary and alternative medicine in various disorders. The present study was undertaken for phytoconstituents estimation and to evaluate chemical compositions of methanolic extract of Withania somnifera (L.) Dunal (Ashwagandha) roots collected from BAU, Ranchi, Jharkhand and Sabour, Bhagalpur, Bihar. Phytochemicals through crude methanolic extract were analyzed in Ashwagandha roots. Alcoholic extract was prepared using Soxhlet extraction apparatus. On phytochemical evaluation, total phenolic, flavonoid, total tannin and alkaloids were estimated. Presence of bioactive components in extract was determined by Liquid Chromatography-Mass Spectrophotometry (ESI positive ion) technique. Qualitative screening of extract ensured the presence of alkaloids, phenols, tannins and flavonoids. Quantitatively these phytochemicals showed TPC (66.06 mg/gm), TFC (650.0 mg/gm), TAC (102 mg/gm) and TTC (5.89 mg/gm). However, the phytochemicals and yield did not accumulate in various fractions on polarity. This result was supported by LC-MS analysis which showed 17 definite peaks in their chromatogram but only 8 peaks offer a suitable approximation. Extract revealed forty five bioactive compounds where most of them were phenolic compounds, aryl propionic acid and cinnamide derivatives. Quantitative analysis of present phytochemical showed maximum quantity of TFC followed by TAC, TPC and TTC. On the other hand, presence of various bioactive compounds in root extract showed that plant is having very high medicinal value.

Purpose: To evaluate the outcomes of water-soluble intrastromal natamycin (IS-NTM) as an adjunct therapy for recalcitrant fungal keratitis. Methods: This was a prospective interventional pilot study in the setting of a tertiary eye-care center. Twenty eyes of 20 consecutive patients with microbiologically proven recalcitrant fungal keratitis (ulcer size >2 mm, depth >50%, and not responding to topical NTM for 2 weeks) were recruited. The selected patients were injected with a novel composition of IS-NTM (10 ug/0.1 mL, soluble natamycin) prepared aseptically in the ocular pharmacology department. All the patients continued using topical NTM suspension 5% 4-hourly until the ulcer healed. Repeat injections were undertaken after 72 h depending on the clinical response and all the patients were followed till 6 months. Results: The mean age of the patients was 40.42 ± 10.09 years. The mean duration of the presentation was 20.8 ± 5.1 days. The most commonly isolated organisms were Aspergillus sp. (12/20, 60%) and Fusarium sp. (8/20, 40%). No patient had iatrogenic perforation or precipitate formation after IS-NTM injection. The overall cure rate with IS-NTM was 95% (19/20 patients). The number of patients who healed with the 1st, 2nd, and 3rd injection was 13, 5, and 1, respectively. One (5%) had no response to treatment and was subjected to penetrating keratoplasty. The average time taken for the resolution of the epithelial defect, stromal infiltrates, and hypopyon was 34 ± 5.2 days, 35.3 ± 6.4 days, and 15 ± 2.5 days. Healing with deep vascularization and cataract was noted in 6/19 eyes (31%) and 13/19 eyes (68.42%), respectively. Conclusion: Intrastromal injection of a novel formulation of NTM holds a promising role as adjunctive therapy to topical NTM in the management of recalcitrant filamentous fungal keratitis. The preliminary results are encouraging and further studies are required to validate the results.

Gunjan Saluja,1 Namrata Sharma,1 Rinky Agarwal,1 H P Sharma,2 Deepali Singhal,1 Prafulla Kumar Maharana,1 Rajesh Sinha,1 Tushar Agarwal,1 T Velpandian,2 Jeewan S Titiyal,1 Gita Satpathy3 1Department of Ophthalmology, Dr. Rajendra Prasad Centre for Ophthalmic Sciences, All India Institute of Medical Sciences, New Delhi, India; 2Ocular Pharmacology and Pharmacy Division, Dr. Rajendra Prasad Centre for Ophthalmic Sciences, All India Institute of Medical Sciences, New Delhi, India; 3Department of Ocular Microbiology, Dr. Rajendra Prasad Centre for Ophthalmic Sciences, All India Institute of Medical Sciences, New Delhi, IndiaCorrespondence: Namrata SharmaDepartment of Ophthalmology, Cornea, Cataract and Refractive Surgery Services, 4th Floor Dr. Rajendra Prasad Centre for Ophthalmic Sciences, All India Institute of Medical Sciences, Ansari Nagar East, New Delhi, Tel +91-11-26593144Fax +91-11-26588919Email namrata.sharma@gmail.comObjective: To compare the safety and efficacy of intrastromal voriconazole (IS-VCZ), amphotericin B (IS-AMB) and natamycin (IS-NTM) as an adjunct to topical natamycin (NTM) in cases of recalcitrant fungal keratitis.Design: Prospective randomized trial.Setting: Tertiary eye centre.Participants: Sixty eyes of 60 patients with microbiologically proven recalcitrant fungal keratitis (ulcer size > 2 mm, depth > 50% of stroma, and not responding to topical NTM therapy for two weeks) were recruited.Methods: patients were randomized into three groups of 20 eyes, each receiving ISVCZ 50ug/0.1 mL, ISAMB, 5ug/0.1 mL and ISNTM 10ug/0.1 mL (prepared aseptically in ocular pharmacology). The patients in all three groups continued topical NTM 5% every four hours until the ulcer healed. Primary outcome measure was time taken till complete clinical resolution of infection, and secondary outcome measure was best corrected visual acuity (BCVA) at six months.Results: All three groups had comparable baseline parameters. The mean duration of healing was significantly better (p=0.02) in the ISNTM group (34± 5.2 days) as compared to the ISVCZ group (36.1± 4.8 days) and the ISAMB group (39.2± 7.2 days). About 95%, 90% and 95% patients healed successfully in the ISVCZ, ISAMB and ISNTM groups, respectively. In terms of healing, deep vascularization was significantly greater in the ISAMB group (55%, p=0.02) when compared to the ISVCZ and ISNTM groups (31% and 26%, respectively). There were fewer repeat injections in the ISNTM group (7/20 vs 8/20 and 9/20 in the ISVCZ and ISNTM groups, respectively).Conclusion: Intrastromal injections are a safe and effective adjunct to conventional therapy in the management of recalcitrant fungal keratitis. ISNTM had a similar visual outcome with faster healing while ISAMB had a higher rate of deep vascularization after healing.Keywords: intrastromal injections, recalcitrant fungal keratitis

Purpose: Purpose of the current study was to assess the presence and functionality of the nucleoside transporters in the lacrimal gland for the tear disposition of its substrate given intravenously...

Background: Plants in either raw form or their root extract constituents utilized as complementary and alternative medicine in various disorders. The present study was undertaken for phytoconstituents estimation and to evaluate chemical compositions of methanolic extract of Withania somnifera (L.) Dunal (Ashwagandha) roots collected from BAU, Ranchi, Jharkhand and Sabour, Bhagalpur, Bihar. Methods: Phytochemicals viz. crude methanolic extract analyzed in Ashwagandha roots. Alcoholic extract were prepared using Soxhlet extraction apparatus. On phytochemical evaluation, total phenolic, flavonoid, total tannin and alkaloids were estimated using their respective methods. Presences of bioactive components in extract were determined by Liquid Chromatography-Mass Spectrophometry (ESI positive ion) technique. Results: Qualitative screening of extract ensured the presence of alkaloids, phenols, tannins and flavonoids. Quantitatively these phytochemicals showed TPC (66.06 mg/gm), TFC (650.0 mg/gm), TAC (102 mg/gm) and TTC (5.89 mg/gm). However, the phytochemicals and yield did not accumulate in various fractions on polarity. This result was supported by LC-MS analysis which showed 17 definite peaks in their chromatogram but only 8 peaks offer a suitable approximation. Extract revealed forty five bioactive compounds where most of them were withanolides, phenolic compounds, aryl propionic acid and cinnamide derivatives. Conclusion: Quantitative analysis of present phytochemical showed maximum quantity of TFC followed by TAC, TPC and TTC. In other hand, presence of various bioactive compounds in root extract showed plant contains very high medicinal value.

The current study was conducted to explore the potential of rutin in preventing sight-threatening diabetic retinopathy. Wistar albino rats (either sex) weighing 200–225 g were intraperitoneally injected with 45 mg/kg streptozotocin (pH 4.5). Rats having blood glucose ≥ 300 mg/dL were divided into two groups (n = 8; each group). Group I served as diabetic control and received normal saline p.o. Group II received rutin 50 mg/kg p.o. for 24 weeks. At the end of 24 weeks, retinal fundus and fluorescein imaging were done, rats were killed, and retinal biochemical assessments were conducted. Moreover, ocular pharmacokinetics of rutin was assessed in the normal rats after a single oral dose of 50 mg/kg. Rutin treatment significantly (p

Objectives Mannose-binding lectin (MBL), an essential innate immune molecule, enhances the opsonization process and activates the complement system. Genetic variations at the promoter and coding region of the MBL-2 gene have been associated with susceptibility to systemic lupus erythematosus (SLE); however, reports remained inconsistent. The present study performs a meta-analysis of published peer-reviewed articles to draw a definitive conclusion. Materials and Methods Published peer-reviewed articles on the association of MBL-2 gene polymorphisms and SLE were screened on various databases such as PubMed (Medline), ScienceDirect, and Google Scholar. A total of 23 eligible articles were included in the present study, comprising 3074 SLE patients and 3985 controls. Genotype and/or allele data for MBL-2 polymorphisms (A > B, A > C, A > D, A > O, Y > X and H > L) were extracted and analyzed by Comprehensive Meta-Analysis software (CMA V3.1). Results The overall analysis revealed a significant association of MBL-2 (A > O) polymorphism with a predisposition to SLE in allele contrast ( p = 0.000; OR = 1.261), homozygous ( p = 0.005; OR = 1.482), heterozygous ( p = 0.004; OR = 1.247), dominant ( p = 0.000; OR = 1.303) and recessive ( p = 0.025; OR = 1.356) genetic comparison model. Similar results were also observed in the comparison of allele and the dominant genetic model of MBL-2 (A > B) polymorphism in overall (allele: p = 0.000, OR = 1.46, dominant: p = 0.001, OR = 1.31) and in the Asian cohorts (allele: p = 0.007, OR = 1.43, dominant: p = 0.008, OR = 1.32). Interestingly, MBL-2 (Y-221X) polymorphism exhibited protection against the development of SLE in heterozygous ( p = 0.005, OR = 0.619) and dominant genetic comparison ( p = 0.01, OR = 0.672) models. Conclusions MBL-2 variants (A > O and A > B) are associated with predisposition to SLE. Conversely, promoter polymorphism (Y-221X) offers protection against SLE development.

Supplemental material, sj-pdf-6-lup-10.1177_0961203320939156 for Association of MBL-2 gene polymorphisms with systemic lupus erythematosus: an updated meta-analysis and trial sequential analysis by Harishankar Mahto, Abhijit Pati, Sushil K Sahu, Hanuman Prasad Sharma, Archana Padhi and Aditya K Panda in Lupus

Supplemental material, sj-pdf-4-lup-10.1177_0961203320939156 for Association of MBL-2 gene polymorphisms with systemic lupus erythematosus: an updated meta-analysis and trial sequential analysis by Harishankar Mahto, Abhijit Pati, Sushil K Sahu, Hanuman Prasad Sharma, Archana Padhi and Aditya K Panda in Lupus

Supplemental material, sj-pdf-1-lup-10.1177_0961203320939156 for Association of MBL-2 gene polymorphisms with systemic lupus erythematosus: an updated meta-analysis and trial sequential analysis by Harishankar Mahto, Abhijit Pati, Sushil K Sahu, Hanuman Prasad Sharma, Archana Padhi and Aditya K Panda in Lupus

Supplemental material, sj-pdf-2-lup-10.1177_0961203320939156 for Association of MBL-2 gene polymorphisms with systemic lupus erythematosus: an updated meta-analysis and trial sequential analysis by Harishankar Mahto, Abhijit Pati, Sushil K Sahu, Hanuman Prasad Sharma, Archana Padhi and Aditya K Panda in Lupus

Supplemental material, sj-pdf-7-lup-10.1177_0961203320939156 for Association of MBL-2 gene polymorphisms with systemic lupus erythematosus: an updated meta-analysis and trial sequential analysis by Harishankar Mahto, Abhijit Pati, Sushil K Sahu, Hanuman Prasad Sharma, Archana Padhi and Aditya K Panda in Lupus

Supplemental material, sj-pdf-3-lup-10.1177_0961203320939156 for Association of MBL-2 gene polymorphisms with systemic lupus erythematosus: an updated meta-analysis and trial sequential analysis by Harishankar Mahto, Abhijit Pati, Sushil K Sahu, Hanuman Prasad Sharma, Archana Padhi and Aditya K Panda in Lupus

Supplemental material, sj-pdf-5-lup-10.1177_0961203320939156 for Association of MBL-2 gene polymorphisms with systemic lupus erythematosus: an updated meta-analysis and trial sequential analysis by Harishankar Mahto, Abhijit Pati, Sushil K Sahu, Hanuman Prasad Sharma, Archana Padhi and Aditya K Panda in Lupus

The objective of the current study was to characterize and evaluate the functional importance of the Nucleoside Transporters (NTs) in the cornea of the rabbits. Reverse transcriptase polymerase chain reaction (RT-PCR) was used for the molecular characterization of the NTs. Their functionality was evaluated using two substrates, ribavirin and cytarabine. Dipyridamole was used as a blocker for the study. All the treatments were given topically. Molecular characterization of NTs revealed presence of ent1, ent2, ent3 and cnt3 in the cornea. The concentration vs time profile for cytarabine in Aqueous Humor (AH) exhibited a statistically significant (p

The purpose of this study was to develop, characterize and evaluate novel water soluble formulation for its topical and intrastromal application. Natamycin complexed with cyclodextrin was characterized by Fourier-transform infrared spectroscopy (FTIR) and differential scanning calorimetry (DSC). The complexed powder was used to formulate 1% w/v aqueous natamycin formulation (Natasol 1%) for topical use. The developed formulation was subjected to stability testing at various conditions. Single and multi-dose trans-corneal permeation of Natasol 1% was evaluated in New Zealand Albino rabbits in comparison with marketed 5% natamycin suspension. Sterile unpreserved Natasol (0.01% w/v natamycin) formulation was also developed for intrastromal injection. Both formulations were evaluated for the ocular toxicity. FTIR and DSC studies revealed successful complexation of natamycin that further protected the degradation at various stability conditions. Single dose trans-corneal permeation studies revealed that Natasol 1% attained Cmax at one hr and maintain its intraocular concentration 5 and 2.5 times higher at 4th and 6th hr compared to 5% natamycin suspension. Multi-dose kinetics revealed the steady state pharmacokinetics after hourly and two hourly dosing schedules. Topical Natasol 1% and sterile unpreserved Natasol 0.01% intrastromal injection, did not show any ocular toxicity in the animals. To conclude, the newly developed topical 1% w/v natamycin formulation was found to be non-inferior to 5% natamycin topical suspension. It is expected to increase patient compliance for the treatment of fungal keratitis.

In the practice of ophthalmology and optometry, several agents are important for therapeutic, cosmetic and diagnostic purposes and cannot be grouped into a single category. They are governed by pharmacopeia or abide by regulatory requirements which are commercially available except few of the customized formulations compounded by hospital pharmacies. This chapter includes most of the formulations extensively used in routine ophthalmic practices like corneal preservative media used to store cornea for transplantation, tattooing solutions for cosmetic use in corneal opacity, agents used in acid or alkali burn, vitreous substitutes, vitreolytic agents, viscoelastic substances, topical immunosuppressants, local anesthetics, botulinum toxin, protease inhibitors, chelators, contact lens solutions and irritation solutions.

The developing visual circuitry attains its mature adult pattern through the process of activity-dependent refinement in which photic stimulation plays the major role. However, auditory stimulation can also facilitate the developing visual Wulst synaptic plasticity and postnatal perceptual behavior, though the underlying mechanism is unclear. We exposed the fertilized eggs of white Leghorn chickens during incubation to either species-specific calls or no sound for varying time periods depending on the functional development of the auditory and/or visual systems. The visual evoked potential (VEP) from the Wulst was recorded at embryonic days (E) 19, 20 and posthatch days (PH) 1-3, to assess functional maturation. A significant attenuation in latencies and higher amplitudes at PH1-3 in the stimulated groups that received exposure during visual system maturation, suggest beneficial effect of auditory inputs only during critical periods. Concomitant with this, there was a significant increase in the expression of BDNF and levels of neurotransmitters GABA, glutamate, norepinephrine and serotonin from E18 only in both hemispheres of the visual Wulst. A significant inter-hemispheric difference in expression was also found in all groups. These results suggest the role of BDNF in activity driven structural and functional maturation of the visual system following prenatal repetitive auditory stimulation.

Background The polyherbal eye drop (Itone™) is a mixture of aqueous distillates of nineteen traditionally used ingredients that sum up to impart potency to the formulation and make it a useful adjunct in various ocular pathologies. However, as there have been no controlled experimental studies accounting to the above claim, therefore, the present study was designed to evaluate the polyherbal formulation (PHF) for antiangiogenic, anti-inflammatory, anticataract, antioxidant and cytotoxicity in addition to the evaluation of intraocular penetration of PHF in rabbit eyes using LC-MS/MS. Materials and methods Antiangiogenic activity of the PHF was evaluated using in ovo chick chorio-allantoic membrane (CAM) assay and in vivo cautery induced corneal neovascularization assay in rats. Anticataract potential was evaluated using steroid induced cataract in developing chick embryos, sodium selenite induced cataract in rat pups and galactose induced cataract in rats. The antioxidant activity was evaluated using di-phenyl picryl hydrazyl (DPPH) radical scavenging assay. Anti-inflammatory activity was evaluated in vitro using inhibition of LTB4 formation in human WBCs and in vivo using carrageenan induced paw edema assay in rats. The cytotoxicity was evaluated against HeLa cancer cell lines using (3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay. Furthermore evaluation of the intraocular penetration of the PHF was carried out in rabbit eyes via aqueous humor paracentesis and further analysis using LC-MS/MS. Results PHF significantly inhibited VEGF induced proliferation of new blood vessels in CAM assay and inhibited the cautery induced corneal neovascularization in rats. Additionally, PHF showed noticeable delay in the progression of cataract in the selenite and galactose induced cataract models whereby the PHF treated lenses were graded for stages II and III respectively. However, the PHF did not show any anticataract activity in the hydrocortisone induced cataract model. Moreover, PHF exhibited anti-inflammatory activity whereby it showed 39.34% inhibition of LTB4 formation and significantly inhibited carrageenan induced paw edema in rats. Eight compounds of PHF viz. camphor, casticin, curcumin-II, quercetin, rosmarinic acid, γ-terpinene, β-pinene and dipentene exhibited transcorneal penetration in rabbit eyes. Conclusion The significant antiangiogenic and anti-inflammatory activities evinced by the PHF merits further investigation for ocular neovascular and inflammatory diseases in humans.