Biometry-Based Technique for Determining the Anterior Scleral Thickness: Validation Using Optical Coherence Tomography Landmarks.

Purpose: Considering the potential role of anterior scleral thickness (AST) in myopia and the ubiquitous use of optical biometers, we applied and validated a biometry-based technique for estimating AST using optical coherence tomography (OCT) landmarks. Methods: The AST was determined across four meridians in 62 participants (aged 20-37 years) with a swept-source OCT and a noncontact optical biometer at a mean ± SD distance of 3.13 ± 0.88 mm from the limbus. The biometer's graticule was focused and aligned with the anterior scleral reflex, which led to the generation of four prominent A-scan peaks: P1 (anterior bulbar conjunctiva), P2 (anterior episclera), P3 (anterior margin of anterior sclera), and P4 (posterior margin of anterior sclera), which were analyzed and compared with the corresponding OCT landmarks to determine tissue thickness. Results: The AST measurements between biometer and OCT correlated for all meridians (r ≥ 0.70, overall r = 0.82; coefficient of variation [CV], 9%-12%; P < 0.01). The mean difference ± SD between two instruments for overall AST measures was 3 ± 2.8 µm (range, -18 to +16 µm; lower limits of agreement, -89 to +83 µm; P = 0.23) across all meridians. The mean ± SE AST with both instruments was found to be thickest at the inferior (562 ± 7 µm and 578 ± 7 µm) and thinnest at the superior (451 ± 7 µm and 433 ± 6 µm) meridian. The biometer demonstrated good intrasession (CV, 8.4%-9.6%) and intersession (CV, 7.9%-13.3%) repeatability for AST measurements across all meridians. Conclusions: The noncontact optical biometer, which is typically used to determine axial length, is capable of accurately estimating AST based on OCT landmarks. Translational Relevance: The high-resolution optical biometers can demonstrate wider application in the field of myopia research and practice to determine AST.

Exploring antiviral potency of N-1 substituted pyrimidines against HIV-1 and other DNA/RNA viruses: Design, synthesis, characterization, ADMET analysis, docking, molecular dynamics and biological activity.

A novel series of pyrimidine derivatives, bearing modified benzimidazoles at N-1 position, has been designed, synthesized and screened as NNRTIs against HIV and as broad-spectrum antiviral agents. The molecules were screened against different HIV targets using molecular docking experiment. The docking results indicated that the molecules interacted well with the residues Lys101, Tyr181, Tyr188, Trp229, Phe227 and Tyr318 present in NNIBP of HIV-RT protein, formed quite stable complexes and, thus, behaved as probable NNRTIs. Among these compounds, 2b and 4b showed anti-HIV activity with IC50 values as 6.65 µg/mL (SI = 15.50) and 15.82 µg/mL (SI = 14.26), respectively. Similarly, compound 1a showed inhibitory property against coxsackie virus B4 and compound 3b against different viruses. Molecular dynamics simulation results unequivocally demonstrated the higher stability of the complex HIV-RT:2b than the HIV-RT:nevirapine complex. The MM/PBSA-based binding free energy (-) 114.92 kJ/mol of HIV-RT:2b complex in comparison to that of HIV-RT:nevirapine complex (-) 88.33 kJ/mol, further demonstrated the higher binding strength of 2b and thus, established the potential of compound 2b as a lead molecule as an HIV-RT inhibitor.

Designing lisuride intranasal nanocarrier system for reduction of oxidative damage with enhanced dopamine level in brain for Parkinsonism.

In the present study, nanoemulsion (NE) loaded with lisuride were formulated for delivering drug to brain via intranasal route. Dopamine levels, pharmacokinetic, and antioxidant activity were estimated. Antioxidant effect of lisuride NE was assessed in-vivo using oxidative stress models revealing symptoms like those of Parkinson's disease. Intranasally administered lisuride NE-treated group revealed a greater number of antioxidant enzymes, such as superoxide dismutase (SOD) and glutathione (GSH) as compared to the intravenously administered lisuride suspension in haloperidol rat model. Additionally, it was observed that lisuride NE can decrease dopamine loss. When lisuride NE was administered intranasally resulted in considerably higher dopamine concentrations (17.48 ± 0.05 ng/mL) in comparison to rats receiving haloperidol (7.28 ± 0.02 ng/mL). From study, it is suggested that NE is a possible strategy to deliver lisuride intranasally to lower free radical damage and prevent the biochemical alterations associated with Parkinson's disease.

Association between relative peripheral refraction and corresponding electro-retinal signals.

PURPOSE: Considering the potential role of the peripheral retina in refractive development and given that peripheral refraction varies significantly with increasing eccentricity from the fovea, we investigated the association between relative peripheral refraction (RPR) and corresponding relative peripheral multifocal electroretinogram (mfERG) responses (electro-retinal signals) from the central to the peripheral retina in young adults. METHODS: Central and peripheral refraction using an open-field autorefractor and mfERG responses using an electrophysiology stimulator were recorded from the right eyes of 17 non-myopes and 24 myopes aged 20-27 years. The relative mfERG N1, P1 and N2 components (amplitude density and implicit time) of a mfERG waveform were compared with the corresponding RPR measurements at the best-matched eccentricities along the principal meridians, that is at the fovea (0°), horizontal (±5°, ±10° and ± 25°) and vertical meridians (±10° and ± 15°). RESULTS: The mean absolute mfERG N1, P1 and N2 amplitude densities (nV/deg2 ) were maximum at the fovea in both non-myopes (N1: 57.29 ± 14.70 nV/deg2 , P1: 106.29 ± 24.46 nV/deg2 , N2: 116.41 ± 27.96 nV/deg2 ) and myopes (N1: 56.25 ± 15.79 nV/deg2 , P1: 100.79 ± 30.81 nV/deg2 , N2: 105.75 ± 37.91 nV/deg2 ), which significantly reduced with increasing retinal eccentricity (p < 0.01). No significant association was reported between the RPR and corresponding relative mfERG amplitudes at each retinal eccentricity (overall Pearson's correlation, r = -0.25 to 0.26, p ≥ 0.09). In addition, the presence of relative peripheral myopia or hyperopia at extreme peripheral retinal eccentricities did not differentially influence the corresponding relative peripheral mfERG amplitudes (p ≥ 0.24). CONCLUSIONS: Relative peripheral mfERG signals are not associated with corresponding RPR in young adults. It is plausible that the electro-retinal signals may respond to the presence of absolute hyperopia (and not relative peripheral hyperopia), which requires further investigation.

Buckeye Rot of Tomato in India: Present Status, Challenges, and Future Research Perspectives.

Tomato in India is commonly exposed to various diseases of fungal, bacterial, and viral origin, resulting in substantial yield losses (≥50%). Buckeye rot (caused by Phytophthora nicotianae var. parasitica) is among the major constraints in the successful cultivation of tomato crops in various parts of the world, including the Solan district of Himachal Pradesh state, India. The fruit rot becomes more devastating under high humidity and wet soils. Symptoms generally appear on green fruit as alternate dark- and light-brown concentric rings. The genome size of P. nicotianae var. parasitica is 82 Mb with >23,000 predicted genes. High humidity (>60%) and optimal temperatures (20 to 25°C), along with rainfall (≥10 mm), help to disperse the pathogen because the inoculum reaches the fruit through splashing rain. Sporangia germinate indirectly by producing zoospores at 20 to 25°C or directly via germ tubes at >25°C. In the absence of suitable resistant varieties, no single management practice is sufficient to keep the disease below the economic threshold level; therefore, integration of cultural and chemical methods is preferable. This article aims to focus on the etiology and management challenges of buckeye rot. We recommend innovative disease management strategies such as identification and deployment of resistant cultivars as well as spraying of synthetic chemical fungicides, biocontrol agents, and use of abiotic chemicals that induce resistance for developing sustainable crop production practices.

Electroretinogram responses in myopia: a review.

The stretching of a myopic eye is associated with several structural and functional changes in the retina and posterior segment of the eye. Recent research highlights the role of retinal signaling in ocular growth. Evidence from studies conducted on animal models and humans suggests that visual mechanisms regulating refractive development are primarily localized at the retina and that the visual signals from the retinal periphery are also critical for visually guided eye growth. Therefore, it is important to study the structural and functional changes in the retina in relation to refractive errors. This review will specifically focus on electroretinogram (ERG) changes in myopia and their implications in understanding the nature of retinal functioning in myopic eyes. Based on the available literature, we will discuss the fundamentals of retinal neurophysiology in the regulation of vision-dependent ocular growth, findings from various studies that investigated global and localized retinal functions in myopia using various types of ERGs.

Generation of the Compression-induced Dedifferentiated Adipocytes (CiDAs) Using Hypertonic Medium.

Current methods to obtain mesenchymal stem cells (MSCs) involve sampling, culturing, and expanding of primary MSCs from adipose, bone marrow, and umbilical cord tissues. However, the drawbacks are the limited numbers of total cells in MSC pools, and their decaying stemness during in vitro expansion. As an alternative resource, recent ceiling culture methods allow the generation of dedifferentiated fat cells (DFATs) from mature adipocytes. Nevertheless, this process of spontaneous dedifferentiation of mature adipocytes is laborious and time-consuming. This paper describes a modified protocol for in vitro dedifferentiation of adipocytes by employing an additional physical stimulation, which takes advantage of augmenting the stemness-related Wnt/ß-catenin signaling. Specifically, this protocol utilizes a polyethylene glycol (PEG)-containing hypertonic medium to introduce extracellular physical stimulation to obtain higher efficiency and introduce a simpler procedure for adipocyte dedifferentiation.

Human Zona Pellucida Glycoproteins: Binding Characteristics With Human Spermatozoa and Induction of Acrosome Reaction.

Human zona pellucida (ZP) matrix is composed of four glycoproteins designated as ZP glycoprotein -1 (ZP1), -2 (ZP2), -3 (ZP3), and -4 (ZP4). Mutations in the genes encoding human ZP glycoproteins are one of the causative factors leading to abnormal ZP matrix and infertility in women. Relevance of the human ZP glycoproteins in 'sperm-oocyte' binding has been delineated by using either transgenic animal models expressing human zona proteins or purified native/recombinant human zona proteins. Studies based on the purified native/recombinant human zona proteins revealed that ZP1, ZP3, and ZP4 primarily bind to the capacitated acrosome-intact human spermatozoa whereas ZP2 binds to acrosome-reacted spermatozoa. On the contrary, human spermatozoa binds to the eggs obtained from transgenic mouse lines expressing human ZP2 but not to those expressing human ZP1, ZP3, and ZP4 suggesting that ZP2 has an important role in human 'sperm-oocyte' binding. Further studies using transgenic mouse lines showed that the N-terminus of human ZP2 mediate the taxon-specific human sperm-oocyte binding. Both glycans and protein-protein interactions have a role in human gamete interaction. Further studies have revealed that the purified native/recombinant human ZP1, ZP3, and ZP4 are competent to induce acrosome reaction. Human sperm binds to the mouse transgenic eggs expressing human ZP1-4 instead of mouse ZP1-3 proteins, penetrated the ZP matrix and accumulated in the perivitelline space, which were acrosome-reacted suggesting that human ZP2 in transgenic mouse model also induce acrosome reaction. In humans N-linked glycosylation of zona proteins have been shown to play an important role in induction of the acrosome reaction. Hence in humans, based on studies using transgenic mouse model as well as purified native/recombinant zona proteins, it is likely that more than one zona protein is involved in the 'sperm-oocyte' binding and induction of the acrosome reaction.

miR-27b-5p inhibits BeWo cells fusion by regulating WNT2B and enzyme involved in progesterone synthesis.

PROBLEM: The miRNAs show placenta-specific expression patterns, which alter during pregnancy-related complications. In present study, the role of miR-27b-5p during forskolin-mediated BeWo cells fusion has been investigated. METHOD OF STUDY: The fusion of BeWo cells in response to forskolin treatment (25 µM) was studied by desmoplakin I+II staining. Expression profile of miR-27b-5p by qRT-PCR and its targets HSD3ß1 and WNT2B by qRT-PCR and in Western blot were studied. The effect of overexpression of miR-27b-5p and silencing of HSD3ß1 & WNT2B by siRNA on forskolin-mediated BeWo cells fusion and secretion of hCG and progesterone by ELISA was investigated. RESULTS: Time-dependent down-regulation in the expression of miR-27b-5p in forskolin-treated BeWo cells has been confirmed by qRT-PCR. Overexpression of miR-27b-5p significantly inhibits forskolin-mediated BeWo cells fusion as well as hCG & progesterone secretion. HSD3ß1 and WNT2B were identified as targets of miR-27b-5p and are up-regulated in forskolin-treated BeWo cells. Overexpression of miR-27b-5p in BeWo cells downregulates their expression. Further, luciferase reporter assay revealed that miR-27b-5p directly target expression of both HSD3ß1 and WNT2B. Silencing of both HSD3ß1 and WNT2B leads to a significant reduction in forskolin-mediated BeWo cells fusion with concomitant decrease in the secretion of progesterone or/and hCG. Decrease in forskolin-mediated cells fusion observed in miR-27b-5p mimic transfected BeWo cells could be rescued by the overexpression of both HSD3ß1 and WNT2B. CONCLUSION: These observations suggest that reduced miR-27b-5p in forskolin-treated BeWo cells leads to increased secretion of progesterone and hCG due to loss of repressional control on HSD3ß1 and WNT2B.

Optical tweezer measurements of asymptotic nonlinearities in complex fluids.

This article presents micro-medium-amplitude oscillatory shear (µMAOS), a method to measure the frequency-dependent micromechanical properties of soft materials in the asymptotically nonlinear regime using optical tweezers. We have developed a theoretical framework to extract these nonlinear mechanical properties of the material from experimental measurements and also proposed a physical interpretation of the third-order nonlinearities measured in single-tone oscillatory tests. We validate the method using a well-characterized surfactant solution of wormlike micelles, and subsequently employ this technique to demonstrate that the cytoplasm of a living cell undergoes strain softening and shear thinning when locally subjected to weakly nonlinear oscillatory deformations.

Alkylated benzimidazoles: Design, synthesis, docking, DFT analysis, ADMET property, molecular dynamics and activity against HIV and YFV.

A series of alkylated benzimidazole derivatives was synthesized and screened for their anti-HIV, anti-YFV, and broad-spectrum antiviral properties. The physicochemical parameters and drug-like properties of the compounds were assessed first, and then docking studies and MD simulations on HIV-RT allosteric sites were conducted to find the possible mode of their action. DFT analysis was also performed to confirm the nature of the hydrogen bonding interaction of active compounds. The in silico studies indicated that the molecules behaved like possible NNRTIs. The nature - polar or non-polar and position of the substituent present at fifth, sixth, and N-1 positions of the benzimidazole moiety played an important role in determining the antiviral properties of the compounds. Among the various compounds, 2-(5,6-dibromo-2-chloro-1H-benzimidazol-1-yl)ethan-1-ol (3a) showed anti-HIV activity with an appreciably low IC50 value as 0.386 × 10-5µM. Similarly, compound 2b, 3-(2-chloro-5-nitro-1H-benzimidazol-1-yl) propan-1-ol, showed excellent inhibitory property against the yellow fever virus (YFV) with EC50 value as 0.7824 × 10-2µM.

Cell swelling, softening and invasion in a three-dimensional breast cancer model.

Sculpting of structure and function of three-dimensional multicellular tissues depend critically on the spatial and temporal coordination of cellular physical properties, yet the organizational principles that govern these events, and their disruption in disease, remain poorly understood. Using a multicellular mammary cancer organoid model, here we map in three dimensions the spatial and temporal evolution of positions, motions, and physical characteristics of individual cells. Compared with cells in the organoid core, cells at the organoid periphery and the invasive front are found to be systematically softer, larger and more dynamic. These mechanical changes are shown to arise from supracellular fluid flow through gap junctions, suppression of which delays transition to an invasive phenotype. Together, these findings highlight the role of spatiotemporal coordination of cellular physical properties in tissue organization and disease progression.

EGF-mediated reduced miR-92a-1-5p controls HTR-8/SVneo cell invasion through activation of MAPK8 and FAS which in turn increase MMP-2/-9 expression.

The members of human miR-17-92 cluster are implicated in several cancers and are known to increase cancer cells invasiveness. The present study reports reduced expression of miR-92a-1-5p in EGF treated HTR-8/SVneo trophoblastic cells by NGS and qRT-PCR. Overexpression of miR-92a-1-5p led to significantly reduced EGF-mediated HTR-8/SVneo cells invasion. MAPK8 and FAS were predicted to be miR-92a-1-5p targets, and confirmed to be reduced by qRT-PCR and Western blotting in trophoblast cells overexpressing miR-92a-1-5p. The binding of miR-92a-1-5p to MAPK8 and FAS 3'-UTR was confirmed by Luciferase reporter assay and Rescue assay. EGF increases MMP-2 & MMP-9 expression and reduces TIMP1 expression in HTR-8/SVneo cells. Inhibition of MAPK8 (by SP600125) reduced EGF-mediated MMP-9/TIMP1 ratio and invasion. Similarly, silencing of FAS by siRNA reduced EGF-mediated MMP-2/TIMP1 ratio and invasion. Treatment of HTR-8/SVneo cells with STAT1/3 inhibitors or siRNAs led to loss of EGF-mediated reduction in miR-92a-1-5p levels. Inserting the predicted binding sites of STAT3 present in promoter region of miR-92a-1-5p upstream of Luciferase promoter reduced its expression in presence of STAT3 expression vector. Thus, EGF leads to reduced miR-92a-1-5p expression which may be regulated by STAT1/STAT3 and controls HTR-8/SVneo cells invasion by targeting MAPK8 and FAS, which in turn increases MMP-2/MMP-9 expression.

Effect of Yoga Ocular Exercises on Eye Fatigue.

BACKGROUND: Comfortable working at near and intermediate tasks depend on the efficiency as well as coordination of accommodation and vergence systems. At present, the need for near and intermediate visual tasks has been dramatically increased, requiring prolonged computer- and gazette-related works. It demands excessive working of the extraocular and ciliary muscles. It may cause eye fatigue and other associated asthenopic symptoms. Globally, eye fatigue is one of the most commonly reported conditions in nonpresbyopic population with asthenopic symptoms. It is necessary to get relief from eye fatigue for better near and intermediate tasks. MATERIALS AND METHODS: Thirty-two undergraduate optometry students who were symptomatic based on a validated eye fatigue questionnaire were included after a baseline comprehensive eye examination. Based on the eye fatigue symptoms score, they were equally assigned to a control group and an exercise group with sixteen participants in each. The exercise group performed yoga ocular exercises for up to 6 weeks after which the eye fatigue symptoms were reassessed in both groups. RESULTS: In the exercise group, there was a statistically significant reduction in eye fatigue scores (P = 0.003), whereas the eye fatigue scores showed significant increment in the control group after 6 weeks (P = 0.044). CONCLUSIONS: Yoga ocular exercises reduce the eye fatigue symptoms score by increasing the efficiency of extraocular muscles. Hence, it could be considered as a therapeutic and nonpharmacologic intervention for reducing the eye fatigue and associated asthenopic symptoms.

Compression-induced dedifferentiation of adipocytes promotes tumor progression.

Dysregulated physical stresses are generated during tumorigenesis that affect the surrounding compliant tissues including adipocytes. However, the effect of physical stressors on the behavior of adipocytes and their cross-talk with tumor cells remain elusive. Here, we demonstrate that compression of cells, resulting from various types of physical stresses, can induce dedifferentiation of adipocytes via mechanically activating Wnt/ß-catenin signaling. The compression-induced dedifferentiated adipocytes (CiDAs) have a distinct transcriptome profile, long-term self-renewal, and serial clonogenicity, but do not form teratomas. We then show that CiDAs notably enhance human mammary adenocarcinoma proliferation both in vitro and in a xenograft model, owing to myofibrogenesis of CiDAs in the tumor-conditioned environment. Collectively, our results highlight unique physical interplay in the tumor ecosystem; tumor-induced physical stresses stimulate de novo generation of CiDAs, which feedback to tumor growth.

BST2 regulates interferon gamma-dependent decrease in invasion of HTR-8/SVneo cells via STAT1 and AKT signaling pathways and expression of E-cadherin.

The mechanism by which interferon-gamma (IFN-γ) downregulates trophoblast invasion needs further investigation. Treatment of HTR-8/SVneo cells with IFN-γ led to a decrease in their invasion concomitant with an increased expression of BST2. Silencing of BST2 by siRNA showed a significant increase in their invasion and spreading after treatment with IFN-γ as well as downregulated expression of E-cadherin. Further, STAT1 silencing inhibited the IFN-γ-dependent increase in the expression of BST2 and E-cadherin. Treatment of HTR-8/SVneo cells with IFN-γ led to the activation of AKT, and its inhibition with PI3K inhibitor abrogated IFN-γ-mediated decrease in invasion/spreading and downregulated BST2 and E-cadherin expression. Collectively, IFN-γ decreases the invasion of HTR-8/SVneo cells by STAT1 and AKT activation via increased expression of BST2 and E-cadherin.

Social mobilization for cholera prevention & control in India: Building on the existing framework.

Social mobilization is an important component of the delivery of vaccines and has to be carried out at different levels. It plays a very critical role in success of a campaign, as was shown by the Polio eradication program in India that was supported by SMNet, a platform created for the purpose. Learnings from this has been used for other vaccine deployments in India as well. In addition, there is a social mobilization effort for routine immunization. A guideline for social mobilization was created by UNICEF specifically for cholera vaccine use during Haiti epidemic in 2010. Since there is a need to develop a roadmap for cholera control in India, especially in the known hotspots, and after natural disasters, we suggest a possible strategy that could be built on the existing framework available in India.

Immunogenicity and contraceptive efficacy of recombinant fusion protein encompassing Sp17 spermatozoa-specific protein and GnRH: Relevance of adjuvants and microparticles based delivery to minimize number of injections.

PROBLEM: Requirement of multiple injections of contraceptive vaccines to achieve infertility is one of the important impediments for their application. In the present study, attempts have been made to reduce the number of injections of contraceptive vaccine. METHOD OF STUDY: Fusion protein encompassing C-terminus fragment of sperm protein Sp17 (aa residues 76-126) and two copies of gonadotropin-releasing hormone along with T-cell epitopes and dilysine linkers (abbreviated as Sp17C -GnRH2 ) was expressed in Escherichia coli. Its immunogenicity and contraceptive efficacy have been evaluated in female FVB/J mice using different adjuvants and delivery platforms. RESULTS: Immunization of female mice with recombinant Sp17C -GnRH2 (25 µg/injection/mouse) emulsified with squalene-arlacel A following two injections schedule led to failure of 88.8% immunized animals to conceive, which was not significantly different from mice immunized with same protein along with alum following three injections schedule. To make single-dose vaccine, poly d,l-lactic acid-based microparticles (PLA-MPs) entrapping Sp17C -GnRH2 were prepared. Immunization of female mice with a combination of soluble Sp17C -GnRH2 (12.5 µg/injection/mouse) along with Sp17C -GnRH2 entrapped in PLA-MPs (12.5 µg/injection/mouse) in alum showed higher antibody titres and contraceptive efficacy as compared to mice immunized with Sp17C -GnRH2 entrapped in PLA-MPs alone in alum. Immunization with recombinant Sp17C -GnRH2 led to long-term infertility as second mating (150 days after immunization) of various groups of immunized mice showed similar infertility as observed during first mating. CONCLUSION: Single-dose immunization with PLA-MPs entrapping Sp17C -GnRH2 along with soluble recombinant protein in alum generated long-lasting infertility in female mice.