Bio-mediated detoxification of heavy metal contaminated soil and phytotoxicity reduction using novel strain of Brevundimonas vancanneytii SMA3.

Heavy metals pose a serious environmental threat on a global scale due to their toxicity towards livings. Therefore, removing harmful metals from the environment has become more challenging in recent years. The objective of this study is to isolate, examine, and characterize naturally existing bacteria that possess the ability to mitigate and detoxify heavy metals such as cadmium, mercury, and lead. The selected bacteria SMA3 actively demonstrated metal tolerance during screening and was then employed for biosorption study using a lab-scale technique. The bacterium belonged to Brevundimonas sp., according to 16 S rRNA analysis. To enhance the removal efficiency of SMA3, response surface methodology (RSM) was employed, resulting in the identification of optimized conditions (pH 7, temperature 30 °C and shaking speed 120 rpm) for achieving maximum removal percentage (69.5 % of Cd, 58.6 % of Hg, and 85.1 % of Pb) within 72 h. The structural changes induced by microbial treatment were demonstrated by comparing the findings of FESEM images and FTIR spectra confirming the disappearance of C ^ C, C]O peaks along with C]O, C-O-C, C-H, and O-H bond destabilization following bioaccumulation. Moreover, in terms of phytotoxicity evaluation, it was observed that the treated soil, containing both heavy metals and the selected potent bacterial strain, exhibited reduced toxicity, resulting in improved germination and growth parameters for the seeds of Solanum lycopersicum (tomato plant). Overall, the selected bacterial strain demonstrated its potential for effectively removing multiple metals from the metal contaminated environment.

3D reconstruction of murine mitochondria reveals changes in structure during aging linked to the MICOS complex.

During aging, muscle gradually undergoes sarcopenia, the loss of function associated with loss of mass, strength, endurance, and oxidative capacity. However, the 3D structural alterations of mitochondria associated with aging in skeletal muscle and cardiac tissues are not well described. Although mitochondrial aging is associated with decreased mitochondrial capacity, the genes responsible for the morphological changes in mitochondria during aging are poorly characterized. We measured changes in mitochondrial morphology in aged murine gastrocnemius, soleus, and cardiac tissues using serial block-face scanning electron microscopy and 3D reconstructions. We also used reverse transcriptase-quantitative PCR, transmission electron microscopy quantification, Seahorse analysis, and metabolomics and lipidomics to measure changes in mitochondrial morphology and function after loss of mitochondria contact site and cristae organizing system (MICOS) complex genes, Chchd3, Chchd6, and Mitofilin. We identified significant changes in mitochondrial size in aged murine gastrocnemius, soleus, and cardiac tissues. We found that both age-related loss of the MICOS complex and knockouts of MICOS genes in mice altered mitochondrial morphology. Given the critical role of mitochondria in maintaining cellular metabolism, we characterized the metabolomes and lipidomes of young and aged mouse tissues, which showed profound alterations consistent with changes in membrane integrity, supporting our observations of age-related changes in muscle tissues. We found a relationship between changes in the MICOS complex and aging. Thus, it is important to understand the mechanisms that underlie the tissue-dependent 3D mitochondrial phenotypic changes that occur in aging and the evolutionary conservation of these mechanisms between Drosophila and mammals.

A Simple, Rapid, and Effective Heparinase Protocol to Enable Nucleic Acid Study from Frozen Heparinized Plasma.

Cell-free RNAs (cfRNAs) are promising analytes as non-invasive biomarkers and have even greater potential if tied in with metabolomics. Plasma is an optimal source for cfRNAs but is often derived from a variety of anticoagulants. Plasma obtained in heparin is suitable for metabolomics but is difficult to utilize for qPCR-based downstream analysis. In the present study, we aimed to develop a simple, time-efficient, and cost-effective heparinase protocol, followed by library preparation and sequencing of human plasma cfRNAs drawn and stored in heparin at -80 °C for several years. Blood was collected in CPT™ sodium heparin tubes from patients with chronic HCV infection (NCT02400216) at the National Institutes of Health (NIH) Clinical Center. Plasma cfRNAs were treated with heparinase I and used for library preparation and next-generation sequencing (NGS). Heparinase treatment maintained RNA integrity and allowed for successful library preparation for all the study subjects even with 7 ng of cfRNAs as starting material. The classification report derived from Pavian R package v1.2.0 showed no artificial reads. The abundance of chordate over microbial reads suggests no addition of experimental error through heparinase I treatment. We report a novel and practical approach to heparinase treatment for human plasma collected and frozen in sodium heparin for several years. This is an effective demonstration of utilizing heparin plasma for NGS and downstream transcriptomic research, which could then be integrated with metabolomics from the same samples, maximizing efficiency and minimizing blood draws.

Non-selective dampening of the host immune response after hepatitis C clearance and its association with circulating chemokine and endotoxin levels.

BACKGROUND & AIMS: Direct-acting antiviral (DAA) therapy has revolutionized treatment for the hepatitis C virus (HCV). While DAA therapy is common, little is known about the intrahepatic immunological changes after sustained virologic response (SVR). We aim to describe transcriptional alterations of the gut microbiome and the liver after SVR. METHODS: Twenty-two HCV patients were evaluated before and 9 months after 12 weeks of sofosbuvir/velpatasvir treatment. All achieved SVR. A liver biopsy, portal blood (direct portal vein cannulation), peripheral blood and stool samples were obtained. RNA-seq and immunofluorescent staining were performed on liver biopsies. RNA-seq and 16S rRNA metagenomics were performed on stool. RESULTS: Differential expression within liver transcription showed 514 downregulated genes (FDR q < .05; foldchange > 2) enriched in inflammatory pathways; of note, GO:0060337, type 1 IFN signalling (p = 8e-23) and GO:0042742, defence response to bacterium (p = 8e-3). Interestingly, microbial products increased in the portal blood and liver after SVR. Due to the increase in microbial products, the gut microbiome was investigated. There was no dysbiosis by Shannon diversity index or Bacteroides/Firmicutes ratio. There was a differential increase in genes responsible for bacterial lipopolysaccharide production after SVR. CONCLUSIONS: The decrease in the antiviral interferon pathway expression was expected after SVR; however, there was an unanticipated decrease in the transcription of genes involved in recognition and response to bacteria, which was associated with increased levels of microbial products. Finally, the alterations in the function of the gut microbiome are a promising avenue for further investigation of the gut-liver axis, especially in the context of the significant immunological changes noted after SVR.

Elicitation of E-waste (acrylonitrile-butadiene styrene) enriched soil bioremediation and detoxification using Priestia aryabhattai MGP1.

Given the rise in both usage and disposal of dangerous electronics, there is a catastrophic rise in assemblage of electronic waste (e-waste). E-waste including various plastic resins are among the most frequently discarded materials in electronic gadgets. In current digital era, managing e-waste has become universal concern. From the viewpoint of persisting lacuna of e-waste managing methods, the current study is designed to fabricate an eco-friendly e-waste treatment with native soil bacteria employing an enrichment culture method. In the presence of e-waste, indigenous soil microbes were stimulated to degrade e-waste. Microbial cultures were isolated using enrichment medium containing acrylonitrile-butadiene styrene (ABS) as the primary carbon source. Priestia aryabhattai MGP1 was found to be the most dominant e-polymer degrading bacterial isolate, as it was reported to degrade ABS plastic in disposed-off television casings. Furthermore, to increase degradation potential of MGP1, Response Surface Methodology (RSM) was adopted which resulted in optimized conditions (pH 7, shaking-speed 120 rpm, and temperature 30 °C), for maximum degradation (18.88%) after 2 months. The structural changes induced by microbial treatment were demonstrated by comparing the findings of Field emission scanning electron microscopy (FESEM) images and Fourier Transform Infrared (FTIR) spectra confirming the disappearance of ≡ C─H peaks along with C-H, C=C and C ≡N bond destabilization following degradation. Energy-dispersive X-ray (EDX) analyzers of the native and decomposed e-polymer samples revealed a considerable loss in elemental weight % of oxygen by 8.4% and silica by 0.5%. Magnesium, aluminium and chlorine which were previously present in the untreated sample, were also removed after treatment by the bacterial action. When seeds of Vigna radiata were screened using treated soil in the presence of both e-waste and the chosen potent bacterial strain, it was also discovered that there was reduced toxicity in terms of improved germination and growth metrics as a phytotoxicity criterion.

Vessel density changes in choroid, chorio-capillaries, deep and superficial retinal plexues on OCTA in normal ageing and various stages of age-related macular degeneration.

PURPOSE: To study vessel density (VD) on optical coherence tomography angiography at choroid, chorio-capillaries (CC) and various retinal levels in normal population and various stages of dry AMD and how these changes progress with increase in severity of the disease. METHODS: Prospective, observational, cross-sectional study was done on 252 eyes of 132 patients (males: 61, females: 71) presenting to tertiary-care centre in Central India between February 2021 and January 2022. For study purpose, eyes were divided into five groups according to the size and number of the drusen, viz, Group 1: No AMD (< 50 years), Group 2: No AMD (> 50 years), Group 3: Early AMD, Group 4: Intermediate AMD and Group 5: Advanced AMD. In all eyes, VD was measured at choroid, CC, deep capillary plexus (DCP) of retina and superficial capillary plexus (SCP) of retina. RESULTS: The mean age in case cohort is 61.90 ± 7.97 years. The mean vascular density differed significantly across diagnosis types in all the quadrants (p < 0.05) at choroid, CC and DCP level. At SCP level, the differences were significant across the groups except at the central quadrant. Vessel density was found to be more in early AMD cohort when compared to No AMD (> 50 years) cohort at SCP and DCP level, while it showed continuous reduction later in intermediate and advanced AMD cohort. CONCLUSION: With increase in the severity of disease, significant reduction in VD is also seen in retinal plexuses, along with the changes in choroid and CC. These VD maps may play a role as non-invasive biomarkers for healthy and diseased ageing.

Homeopathic Medicines in Second Wave of COVID-19: Prognostic Factor Research.

BACKGROUND: The clinical profile and course of COVID-19 evolved perilously in a second wave, leading to the use of various treatment modalities that included homeopathy. This prognostic factor research (PFR) study aimed to identify clinically useful homeopathic medicines in this second wave. METHODS: This was a retrospective, multi-centred observational study performed from March 2021 to May 2021 on confirmed COVID-19 cases who were either in home isolation or at COVID Care Centres in Delhi, India. The data were collected from integrated COVID Care Centres where homeopathic medicines were prescribed along with conventional treatment. Only those cases that met a set of selection criteria were considered for analysis. The likelihood ratio (LR) was calculated for the frequently occurring symptoms of the prescribed medicines. An LR of 1.3 or greater was considered meaningful. RESULTS: Out of 769 confirmed COVID-19 cases reported, 514 cases were selected for analysis, including 467 in home isolation. The most common complaints were cough, fever, myalgia, sore throat, loss of taste and/or smell, and anxiety. Most cases improved and there was no adverse reaction. Certain new symptoms, e.g., headache, dryness of mouth and conjunctivitis, were also seen. Thirty-nine medicines were prescribed, the most frequent being Bryonia alba followed by Arsenicum album, Pulsatilla nigricans, Belladonna, Gelsemium sempervirens, Hepar sulphuris, Phosphorus, Rhus toxicodendron and Mercurius solubilis. By calculating LR, the prescribing indications of these nine medicines were ascertained. CONCLUSION: Add-on use of homeopathic medicines has shown encouraging results in the second wave of COVID-19 in integrated care facilities. Further COVID-related research is required to be undertaken on the most commonly prescribed medicines.

A review on biosurfactant producing bacteria for remediation of petroleum contaminated soils.

The discharge of potentially toxic petroleum hydrocarbons into the environment has been a matter of concern, as these organic pollutants accumulate in many ecosystems due to their hydrophobicity and low bioavailability. Petroleum hydrocarbons are neurotoxic and carcinogenic organic pollutants, extremely harmful to human and environmental health. Traditional treatment methods for removing hydrocarbons from polluted areas, including various mechanical and chemical strategies, are ineffective and costly. However, many indigenous microorganisms in soil and water can utilise hydrocarbon compounds as sources of carbon and energy and hence, can be employed to degrade hydrocarbon contaminants. Therefore, bioremediation using bacteria that degrade petroleum hydrocarbons is commonly viewed as an environmentally acceptable and effective method. The efficacy of bioremediation can be boosted further by using potential biosurfactant-producing microorganisms, as biosurfactants reduce surface tension, promote emulsification and micelle formation, making hydrocarbons bio-available for microbial breakdown. Further, introducing nanoparticles can improve the solubility of hydrophobic hydrocarbons as well as microbial synthesis of biosurfactants, hence establishing a favourable environment for microbial breakdown of these chemicals. The review provides insights into the role of microbes in the bioremediation of soils contaminated with petroleum hydrocarbons and emphasises the significance of biosurfactants and potential biosurfactant-producing bacteria. The review partly focusses on how nanotechnology is being employed in different critical bioremediation processes.

Pre-operative optical coherence tomography predictors: Do they hold any relevance in the era of inverted internal limiting membrane flap in large macular holes?

Purpose: To study the relevance of preoperative OCT predictors in large macular holes (MH) treated using the inverted ILM peel technique. Methods: Prospective study of 95 patients undergoing vitrectomy for large MH between January 2019 and December 2020 was performed by dividing the patients into groups depending on various quantitative parameters and indices of MH such as base diameter (BD), hole form factor (HFF), macular hole index (MHI), diameter hole index (DHI), and tractional hole index (THI) by using parameters such as minimal hole diameter, hole height, nasal and temporal arm lengths. Depending upon the duration of symptoms, patients were divided into three groups: <3 months, 3-6 months, and >6 months. Anatomical success rate, type of closure, and postoperative vision gain were analyzed in relation to the abovementioned diameters, indices, and duration to see if any significance existed. Results: The mean age of patients included in the study was 60.48 ± 13.88 years, with female preponderance (males: females = 37:58). Change in logMAR was statistically significant individually with all studied parameters (P < 0.0001) without influence of size of hole and other indices. BD and DHI levels showed significant association with type of closure as indicated by P values of 0.017 and 0.048, respectively. Duration of symptoms showed no significance in terms of anatomical and functional success. Conclusion: OCT predictors of MH success seem to have lost relevance with inverted flap surgeries as 100% anatomical success is achieved with this technique, with 95.78% (91/95) achieving type 1 closure with statistically significant equivalent functional gain across the indices with no effect of duration of symptoms.

Impact of Bias in Data Collection of COVID-19 Cases.

BACKGROUND: Prognostic factor research (PFR), prevalence of symptoms and likelihood ratio (LR) play an important role in identifying prescribing indications of useful homeopathic remedies. It involves meticulous unbiased collection and analysis of data collected during clinical practice. This paper is an attempt to identify causes of bias and suggests ways to mitigate them for improving the accuracy in prescribing for better clinical outcomes and execution of randomized controlled studies. METHODS: A prospective, open label, observational study was performed from April 2020 to December 2020 at two COVID Health Centers. A custom-made Excel spreadsheet containing 71 fields covering a spectrum of COVID-19 symptoms was shared with doctors for regular reporting. Cases suitable for PFR were selected. LR was calculated for commonly occurring symptoms. Outlier values with LR ≥5 were identified and variance of LRs was calculated. RESULTS: Out of 1,889 treated cases of confirmed COVID-19, 1,445 cases were selected for pre-specified reasons. Nine medicines, Arsenicum album, Bryonia alba, Gelsemium sempervirens, Pulsatilla nigricans, Hepar sulphuricus, Magnesia muriaticum, Phosphorus, Nux vomica and Belladonna, were most frequently prescribed. Outlier values and large variance for Hepar sulphuricus and Magnesia muriaticum were noticed as indication of bias. Confirmation bias leading to lowering of symptom threshold, keynote prescribing, and deficiency in checking of all symptoms in each case were identified as the most important sources of bias. CONCLUSION: Careful identification of biases and remedial steps such as training of doctors, regular monitoring of data, checking of all pre-defined symptoms, and multicenter data collection are important steps to mitigate biases.

Generalizability of Homeopathic Prognostic Factor Research Outcome in COVID-19 Treatment: Comparison of Data.

BACKGROUND/OBJECTIVE: During the coronavirus disease 2019 (COVID-19) pandemic, several homeopathic prognostic factor research (PFR) projects have been undertaken. We found two projects with comparable outcomes to assess consistency and possible flaws. METHODS: Two comparisons were made. (1) Outcome of a PFR data collection from the Liga Medicorum Homoeopathica Internationalis (LMHI) by about 100 doctors with 541 cases was compared with a previous analysis of 161 cases in the same database. (2) The updated LMHI database was also compared with a data collection carried out in India by four doctors with a total of 1,445 cases. Differences that resulted in conflicting outcomes (indication in one, contraindication in the other) were examined for possible causes. RESULTS: There was only a single outcome in the updated LMHI database that conflicted with the previous dataset, and this could have been due to statistical variation. The Indian data contained many cases, from few doctors, while the LMHI database had few cases per doctor, but many doctors. The overlap between the projects (individual cases entered in both) was between zero and 22%. In 72 comparisons we found six (8.3%) conflicting outcomes. Possible causes were statistical error due to small numbers of cases and/or observers, confirmation bias, and keynote prescribing if this resulted in symptoms being inadequately checked. CONCLUSION: There was little conflict between the outcomes of the two versions of one project and between the two different PFR projects. Differences could mostly be explained by causes that can be managed. This consistency should primarily be interpreted as showing a strong overall consensus between homeopathic practitioners worldwide, but with variation of consensus between small groups of practitioners.

Zinc Signaling in the Mammary Gland: For Better and for Worse.

Zinc (Zn2+) plays an essential role in epithelial physiology. Among its many effects, most prominent is its action to accelerate cell proliferation, thereby modulating wound healing. It also mediates affects in the gastrointestinal system, in the testes, and in secretory organs, including the pancreas, salivary, and prostate glands. On the cellular level, Zn2+ is involved in protein folding, DNA, and RNA synthesis, and in the function of numerous enzymes. In the mammary gland, Zn2+ accumulation in maternal milk is essential for supporting infant growth during the neonatal period. Importantly, Zn2+ signaling also has direct roles in controlling mammary gland development or, alternatively, involution. During breast cancer progression, accumulation or redistribution of Zn2+ occurs in the mammary gland, with aberrant Zn2+ signaling observed in the malignant cells. Here, we review the current understanding of the role of in Zn2+ the mammary gland, and the proteins controlling cellular Zn2+ homeostasis and signaling, including Zn2+ transporters and the Gq-coupled Zn2+ sensing receptor, ZnR/GPR39. Significant advances in our understanding of Zn2+ signaling in the normal mammary gland as well as in the context of breast cancer provides new avenues for identification of specific targets for breast cancer therapy.

Digitally assisted three-dimensional surgery - Beyond vitreous.

Purpose: The aim of this study was to evaluate the application and safety of three-dimensional (3D) visualization system in varied anterior segment procedures and Scleral Buckle. Methods: This was a prospective observational study of 313 eyes. Patients undergoing phacoemulsification (PE) with intraocular lens (IOL), trabeculectomies, glaucoma triple procedure (GTP), scleral fixated (SF) IOL, and scleral buckle (SB) were included in the study. Cases were randomly distributed in 3D visualization system (learning and post-learning phase) and conventional microscope group. Parameters studied were complications (intraoperative and early postoperative), surgical outcomes, and surgeon's perspective on various parameters (through a validated questionnaire) like surgical time, time lag, learning curve, ease of doing various steps and its value as an educational tool, for both groups [Questionnaires 1 and 2]. Results: Complications rates were not different in two groups. Surgical outcomes (anatomical and physiological) were similar in both the groups. Mean duration of surgery in PE+IOL, Trabeculectomy, GTP in learning stage by 3D was significantly higher than Microscope, which became insignificant in postlearning stage. For, SB and SFIOL, duration between two groups were insignificantly different. There was significant learning struggle in PE+IOL, SB, and Trabeculectomy. Image resolution, depth perception, illumination and postural comfort was graded higher for 3D surgery across the stages. Time lag, poor color contrast, and field of view were appreciated during the learning stage. Educational relevance of 3D was higher, as appreciated by resident and nurses. Conclusion: 3D surgery is as safe, faster, and predictable after initial learning struggle. Even in anterior segment procedure, no apparent lag was appreciated after learning curve.

First Detection of Batrachochytrium dendrobatidis in Wild Frogs from Bangladesh.

Global amphibian populations are facing a novel threat, chytridiomycosis, caused by the fungus Batrachochytrium dendrobatidis (Bd), which is responsible for the severe decline of a number of species across several continents. Chytridiomycosis in Asia is a relatively recent discovery yet there have been no reports on Bd-presence in Bangladeshi amphibians. We conducted a preliminary study on 133 wild frogs from seven sites in Bangladesh between April and July 2018. Nested PCR analysis showed 20 samples (15.04%) and 50% of the tested taxa (9 species from 6 genera and 4 families) as Bd-positive. Eight of the nine species are discovered as newly infected hosts. Analysis of Bd-positive samples shows prevalence does not significantly vary among different land cover categories, although the occurrence is higher in forested areas. The prevalence rate is similar in high and low disturbed areas, but the range of occurrence is statistically higher in low disturbance areas. Maximum entropy distribution modeling indicates high probabilities of Bd occurrence in hilly and forested areas in southeast and central-north Bangladesh. The Bd-specific ITS1-5.8S-ITS2 ribosomal gene sequence from the Bd-positive samples tested is completely identical. A neighbor-joining phylogenetic tree reveals that the identified strain shares a common ancestry with strains previously discovered in different Asian regions. Our results provide the first evidence of Bd-presence in Bangladeshi amphibians, inferring that diversity is at risk. The effects of environmental and climatic factors along with quantitative PCR analysis are required to determine the infection intensity and susceptibility of amphibians in the country.

Evaluation of ganglion cell-inner plexiform layer thickness in the diagnosis of preperimetric glaucoma and comparison to retinal nerve fiber layer.

Purpose: The aim of this study was to evaluate the diagnostic ability of optic nerve head (ONH), RNFL, and GC-IPL parameters in differentiating eyes with PPG from normals. Methods: This was a retrospective, cross-sectional, observational study. We studied 73 eyes of 41 patients and compared them to 65 eyes of 34 normal persons. Each patient underwent detailed ocular examination, standard automated perimetry, GC-IPL, ONH, and RNFL analysis. PPG was defined as eyes with normal visual field results and one or more localized RNFL defects that were associated with a glaucomatous disc appearance (e.g., notching or thinning of neuroretinal rim) and IOP more than 21 mm Hg. Diagnostic abilities of GC-IPL, ONH, and RNFL parameters were computed using area under receiver-operating curve (AUROC), sensitivity and specificity, and likelihood ratios (LRs). Results: All GC-IPL parameters differed significantly from normal. The ONH, RNFL, and GC-IPL parameters with best area under curves (AUCs) to differentiate PPG were vertical cup to disc ratio (0.76), inferior quadrant RNFL thickness (0.79), and inferotemporal quadrant GC-IPL thickness (0.73), respectively. Similarly, best LRs were found for clock hour 5, 6, and 12 thicknesses among RNFL; inferior sector and inferotemporal sector thicknesses among GC-IPL parameters. Conclusion: Diagnostic abilities of GC-IPL parameters were comparable to RNFL parameters in differentiating PPG patients from normals. The likelihood of ruling in a disease was greater with GC-IPL parameters.

ZnR/GPR39 controls cell migration by orchestrating recruitment of KCC3 into protrusions, re-organization of actin and activation of MMP.

Actin re-organization and degradation of extracellular matrix by metalloproteases (MMPs) facilitate formation of cellular protrusions that are required for cell proliferation and migration. We find that Zn2+ activation of the Gq-coupled receptor ZnR/GPR39 controls these processes by regulating K+/Cl- co-transporter KCC3, which modulates cell volume. Silencing of KCC3 expression or activity reverses ZnR/GPR39 enhancement of cell proliferation, migration and invasion through Matrigel. Activation of ZnR/GPR39 recruits KCC3 into F-actin rich membrane protrusions, suggesting that it can locally control volume changes. Immunofluorescence analysis indicates that Zn2+ activation of ZnR/GPR39 and KCC3 are required to enhance formation of F-actin stress fibers and cellular protrusions. In addition, ZnR/GPR39 upregulation of KCC3-dependent transport increases the activity of matrix metalloproteases MMP2 and MMP9. Our study establishes a mechanism in which ZnR/GPR39 orchestrates localization and activation of KCC3, formation of F-actin rich cell protrusions and activation of MMPs, and thereby controls cell proliferation and migration.

Structural evaluation of perimetrically normal and affected hemifields in open angle glaucoma.

Purpose: To study macular ganglion cell layer--inner plexiform layer complex (GCL + IPL) in relation to peripapillary retinal nerve fiber layer (RNFL) in glaucomatous eyes with superior or inferior hemifield defects (HD) and to study structural configuration in normal hemifield. Methods: This was an observational cross-sectional study. Data from consecutive 45 superior HD (SHD) and 50 inferior HD (IHD) eyes were analyzed. Each patient underwent detailed ocular examination, standard automated perimetry, and spectral domain optical coherence tomography (SD-OCT). After adjusting for age, gender, and signal strength, area under receiver operating characteristic curve (AUC) was calculated to determine diagnostic ability of GCL + IPL and peripapillary RNFL. Apparently normal hemifield was compared with true normal hemifield. Data were analyzed with SPSS, analysis of variance, t-test, Chi-square test, and receiver operating curve. Results: In the SHD glaucoma group, best parameters for discriminating normal eyes from glaucomatous eyes were inferotemporal GCL + IPL thickness (0.935) and inferior quadrant RNFL thickness (0.971). For IHD glaucoma, average GCL + IPL thickness (0.877) and average RNFL thickness (0.950) had best AUC values. When evaluating apparently normal hemifield in both groups, statistically significant difference was found in inferior GCL + IPL sector (0.865) and inferior quadrant RNFL (0.883) in IHD and superonasal GCL + IPL (0.725) and superior quadrant RNFL (0.842) in SHD groups. Conclusion: SD-OCT may be a useful ancillary diagnostic tool for evaluation of early macular and circumpapillary structural changes in glaucomatous eyes with localized visual field defects. Apparently normal hemifields show structural damage and should be considered in management of glaucoma.

Internal limiting membrane peel: Does it change the success rate of primary vitrectomy without belt buckle in rhegmatogenous retinal detachments?

Purpose: To compare the anatomic success of pars plana vitrectomy (PPV) after internal limiting membrane (ILM) peeling at macular area and macular plus peripapillary area versus no peeling in rhegmatogenous retinal detachments (RRD). Methods: A prospective observational study between July 2014 and March 2017 conducted on 289 eyes of 287 patients with RRD were randomly assigned to three treatment procedures, viz., PPV with no ILM peeling, PPV with macular peeling, and PPV with macular plus peripapillary peeling. Recurrent RD (ReRD) was treated as an event and accordingly the overall primary (PS) and final success (FS) rates were obtained. The risk of ReRD associated with peeling procedures after adjusting for risk factors were obtained using Cox-proportional hazard analysis. Results: The PS percentage for no peel, macular, and macular plus peripapillary procedures were 77.78% (70/90), 82.18% (83/101), and 94.89% (93/98; maximum), respectively, which was statistically significant with a P value of 0.003. The FS percentage for no peel, macular, and macular plus peripapillary were 93.33%, 95.04%, and 100%, respectively, which was significantly different with a P value of 0.048. With reference to no peeling, the adjusted hazard ratio for macular peeling was 0.841 [95% CI: 0.44-1.60] while 0.235 [95% CI: 0.088-0.626] for macular plus peripapillary peeling. Conclusion: The anatomic success rate of PPV with macular plus peripapillary ILM peeling was significantly higher as compared to no peel category. The hazard of ReRD in patients undergoing macular plus peripapillary peel was significantly reduced as compared to no peel procedure.

Functional interrelation of MYC2 and HY5 plays an important role in Arabidopsis seedling development.

Arabidopsis MYC2 bHLH transcription factor plays a negative regulatory role in blue light (BL)-mediated seedling development. HY5 bZIP protein works as a positive regulator of multiple wavelengths of light and promotes photomorphogenesis. Both MYC2 and HY5, belonging to two different classes of transcription factors, are the integrators of multiple signaling pathways. However, the functional interrelations of these two transcription factors in seedling development remain unknown. Additionally, whereas HY5-mediated regulation of gene expression has been investigated in detail, the transcriptional regulation of HY5 itself is yet to be understood. Here, we show that HY5 and MYC2 work in an antagonistic manner in Arabidopsis seedling development. Our results reveal that HY5 expression is negatively regulated by MYC2 predominantly in BL, and at various stages of development. On the other hand, HY5 negatively regulates the expression of MYC2 at various wavelengths of light. In vitro and in vivo DNA-protein interaction studies suggest that MYC2 binds to the E-box cis-acting element of HY5 promoter. Collectively, this study demonstrates a coordinated regulation of MYC2 and HY5 in blue-light-mediated Arabidopsis seedling development.

Linear discriminant score for differentiating early primary open angle glaucoma from glaucoma suspects.

PURPOSE: To determine the diagnostic accuracy of a linear discriminant function (LDF) based on macular ganglion cell complex (GCC), optic nerve head (ONH) and retinal nerve fibre layer (RNFL) for differentiating early primary open-angle glaucoma (POAG) from glaucoma suspects. METHODS: In this cross-sectional study, data from consecutive 127 glaucoma suspects and 74 early POAG eyes were analysed. Each patient underwent detailed ocular examination, standard automated perimetry, GCC and ONH and RNFL analysis. After adjusting for age, gender and signal strength using the analysis of covariance; Benjamin-Hochberg multiple testing correction was performed to detect truly significant parameters to calculate the LDF. Subsequently, diagnostic accuracy of GCC and ONH and RNFL were determined. The obtained LDF score was evaluated for diagnostic accuracy in another test set of 32 suspect and 19 glaucomatous eyes. Data were analysed with the R-3.2.1 (R Core Team 2015), analysis of variance, t-test, Chi-square test and receiver operating curve. RESULTS: Among all GCC parameters, infero temporal had the best discriminating power and average RNFL thickness and vertical CDR among ONH and RNFL parameters. LDF scores for GCC had AUROC of 0.809 for a cut-off value 0.07, while scores for ONH and RNFL had AUROC of 0.903 for a cut-off value - 0.24. Analysis on combined parametric space resulted in avg RNFL thickness, vertical CDR, min GCC + IPL and superior GCC + IPL as key parameters. LDF scores obtained had AUROC of 0.924 for a cut-off value 0.1. The LDF was applied to a test set with an accuracy of 84.31%. CONCLUSION: The LDF had a better accuracy than individual GCC and ONH and RNFL parameters and can be used for diagnosis of glaucoma.