Comparative and evolutionary analysis of α-amylase gene across monocots and dicots.

α-amylase is an important enzyme involved in starch degradation to provide energy to the germinating seedling. The present study was conducted to reveal structural and functional evolution of this gene among higher plants. Discounting polyploidy, most plant species showed only a single copy of the gene making multiple isoforms in different tissues and developmental stages. Genomic length of the gene ranged from 1472 bp in wheat to 2369 bp in soybean, and the size variation was mainly due to differences in the number and size of introns. In spite of this variation, the intron phase distribution and insertion sites were mostly conserved. The predicted protein size ranged from 414 amino acid (aa) in soybean to 449aa in Brachypodium. Overall, the protein sequence similarity among orthologs ranged from 56.4 to 97.4 %. Key motifs and domains along with their relative distances were conserved among plants although several species, genera, and class specific motifs were identified. The glycosyl hydrolase superfamily domain length varied from 342aa in soybean to 384aa in maize and sorghum while length of the C-terminal ß-sheet domain was highly conserved with 61aa in all monocots and Arabidopsis but was 59aa in soybean and Medicago. Compared to rice, 3D structure of the proteins showed 89.8 to 91.3 % similarity among the monocots and 72.7 to 75.8 % among the dicots. Sequence and relative location of the five key aa required for the ligand binding were highly conserved in all species except rice.

Employing in silico investigations to determine the cross-kingdom approach for Curcuma longa miRNAs and their human targets.

Background: Plant elements and extracts have been used for centuries to treat a wide range of diseases, from cancer to modern lifestyle ailments like viral infections. These plant-based miRNAs have the capacity to control physiological and pathological conditions in both humans and animals, and they might be helpful in the detection and treatment of a variety of diseases. The present study investigates the miRNA of the well-known spice Curcuma Longa and its prospective targets using a variety of bioinformatics techniques. Results: Using the integrative database of animal, plant, and viral microRNAs known as miRNEST 2.0, nine C. longa miRNAs were predicted. psRNA target service foretells the presence of 23 human target genes linked to a variety of disorders. By interacting with a variety of cellular and metabolic processes, miRNAs 167, 1525, and 756 have been found to be critical regulators of tumour microenvironment. SARS-cov2 and influenza A virus regulation have been connected to ZFP36L1 from miRNA 1525 and ETV5 from miRNA 756, respectively. Conclusions: The current cross-kingdom study offers fresh knowledge about how to increase the effectiveness of plant-based therapies for disease prevention and serves as a platform for in vitro and in vivo research development.

Recent trends in miRNA therapeutics and the application of plant miRNA for prevention and treatment of human diseases.

Background: Researchers now have a new avenue to investigate when it comes to miRNA-based therapeutics. miRNAs have the potential to be valuable biomarkers for disease detection. Variations in miRNA levels may be able to predict changes in normal physiological processes. At the epigenetic level, miRNA has been identified as a promising candidate for distinguishing and treating various diseases and defects. Main body: In recent pharmacology, plants miRNA-based drugs have demonstrated a potential role in drug therapeutics. The purpose of this review paper is to discuss miRNA-based therapeutics, the role of miRNA in pharmacoepigenetics modulations, plant miRNA inter-kingdom regulation, and the therapeutic value and application of plant miRNA for cross-kingdom approaches. Target prediction and complementarity with host genes, as well as cross-kingdom gene interactions with plant miRNAs, are also revealed by bioinformatics research. We also show how plant miRNA can be transmitted from one species to another by crossing kingdom boundaries in this review. Despite several unidentified barriers to plant miRNA cross-transfer, plant miRNA-based gene regulation in trans-kingdom gene regulation may soon be valued as a possible approach in plant-based drug therapeutics. Conclusion: This review summarised the biochemical synthesis of miRNAs, pharmacoepigenetics, drug therapeutics and miRNA transkingdom transfer.

Determination of dideoxyosone precursors of AGEs in human lens proteins.

Dideoxyosones (DDOs) are intermediates in the synthesis of advanced glycation endproducts (AGEs), such as pentosidine and glucosepane. Although the formation of pentosidine and glucosepane in the human lens has been firmly established, the formation of DDOs has not been demonstrated. The aim of this study was to develop a reliable method to detect DDOs in lens proteins. A specific DDO trapping agent, biotinyl-diaminobenzene (3,4-diamino-N-(3-[5-(2-oxohexahydro-1H-thieno[3,4-d]imidazol-4-yl)pentanoyl]aminopropyl)benzamide) (BDAB) was added during in vitro protein glycation or during protein extraction from human lenses. In vitro glycated human lens protein showed strong reaction in monomeric and polymeric crosslinked proteins by Western blot and ELISA. Glycation of BSA in the presence of BDAB resulted in covalent binding of BDAB to the protein and inhibited pentosidine formation. Mass spectrometric analysis of lysozyme glycated in the presence of BDAB showed the presence of quinoxalines at lysine residues at positions K1, K33, K96, and K116. The ELISA results indicated that cataractous lens proteins contain significantly higher levels of DDO than non-cataractous lenses (101.9±67.8 vs. 31.7±19.5AU/mg protein, p<0.0001). This study provides first direct evidence of DDO presence in human tissue proteins and establishes that AGE crosslink synthesis in the human lens occurs via DDO intermediates.

Histomorphological and immunofluorescence evaluation of bimanual and coaxial phacoemulsification incisions in rabbits.

PURPOSE: To compare the changes in the histomorphology and immunofluorescence of collagen type I in clear corneal incisions (CCIs) at the end of bimanual and coaxial phacoemulsification in rabbits. SETTING: Iladevi Cataract and IOL Research Centre, Ahmedabad, India. METHODS: In this randomized study, the left eye of 30 rabbits had bimanual phacoemulsification through 1.2 mm CCIs for phaco tip access accompanied by an additional 1.4 mm incision for irrigating chopper access or coaxial phacoemulsification through a 2.6 mm single-plane CCI. The right eyes acted as controls. Samples collected at the end of surgery were processed for histomorphology using periodic acid-Schiff-hematoxylin staining and immunofluorescence localization of type I collagen. RESULTS: Incisions exposed to bimanual phacoemulsification had loss of epithelium, stromal fiber shrinkage, keratocyte nuclei fragmentation and condensation, ragged tunnel margins, and Descemet membrane and endothelial cell loss. The changes were minimal or absent in incisions exposed to coaxial phacoemulsification. Immunofluorescence showed a loss of parallel arrangement of type I collagen fibers in bimanual phacoemulsification incisions, while the fibers were well preserved in coaxial phacoemulsification incisions. The changes were more prominent in the roof of the incision tunnel than in the floor. CONCLUSIONS: Corneal incisions for bimanual phacoemulsification were more prone to damage than those for coaxial phacoemulsification. This may be attributed to the absence of a sleeve, which places the incision tunnel in direct contact with the metal phaco tip.

The optimisation of electrokinetic remediation for heavy metals and radioactivity contamination on Holyrood-Lunas soil (acrisol species) in Sri Gading Industrial Area, Batu Pahat, Johor, Malaysia.

The optimisation of electrokinetic remediation of an alluvial soil, locally named as Holyrood-Lunas from Sri Gading Industrial Area, Batu Pahat, Johor, Malaysia, had been conducted in this research. This particular soil was chosen due to its relatively high level of background radiation in a range between 139.2 and 539.4 nGy h(-1). As the background radiation is correlated to the amount of parent nuclides, (238)U and (232)Th, hence, a remediation technique, such as electrokinetic, is very useful in reducing these particular concentrations of heavy metal and radionuclides in soils. Several series of electrokinetics experiments were performed in laboratory scale in order to study the influence of certain electrokinetic parameters in soil. The concentration before (pre-electrokinetic) and after the experiment (post-electrokinetic) was determined via X-ray fluorescence (XRF) analysis technique. The best electrokinetic parameter that contributed to the highest achievable concentration removal of heavy metals and radionuclides on each experimental series was incorporated into a final electrokinetic experiment. Here, High Pure Germanium (HPGe) was used for radioactivity elemental analysis. The XRF results suggested that the most optimised electrokinetic parameters for Cr, Ni, Zn, As, Pb, Th and U were 3.0 h, 90 volts, 22.0 cm, plate-shaped electrode by 8 × 8 cm and in 1-D configuration order whereas the selected optimised electrokinetic parameters gave very low reduction of (238)U and (232)Th at 0.23 ± 2.64 and 2.74 ± 23.78 ppm, respectively.

Sequential effects of ultraviolet radiation on the histomorphology, cell density and antioxidative status of the lens epithelium--an in vivo study.

In vivo progressive effects of UV irradiation on the lens epithelium were studied using various histomorphological and biochemical parameters. Fifteen day old rat pups were exposed to 600 mW/m2 of radiation, including UV-A and UV-B, 12 h daily for 90, 120, 150 and 180 days. Biochemical parameters such as protein-bound and non-protein-bound sulfhydryl groups in both soluble and insoluble fractions and enzymes, which play an important role in combating the oxidative stress, were studied. Decreased cell density of lens epithelial cells (LEC) was observed in all three zones along with the decrease in the levels of soluble sulfhydryls (S-SH), glutathione reductase (GR), superoxide dismutase (SOD), glutathione peroxidase (GPx) and catalase (CAT). On the other hand, an increase in insoluble sulfhydryls was observed. Because of the decrease in S-SH and GR activities, the LEC became vulnerable to oxidative stress. Decreased activities of SOD, GPx and CAT suggest elevated oxidative stress. This effect of UV radiation may lead to cell death that may be responsible for the observed decrease in the cell density in all three zones of the lens epithelium.

Epidemiology based etiological study of pediatric cataract in western India.

BACKGROUND: Cataract is responsible for about 10% blindness among children in India. Etiology of cataract is not well defined especially for childhood cataracts and epidemiological data for Indian population is not available in details. AIM: This study was performed to survey the causes of childhood cataracts and to identify the preventable factors in four western states of India. SETTINGS AND DESIGN: The present study is a hospital-based, prospective study on 172 consecutive pediatric cataract patients. MATERIAL AND METHODS: Type of cataract was determined using slit-lamp bio-microscopy or operation microscope after mild general anesthesia especially on very young babies. Other anomalies of eye were determined using appropriate ophthalmic instruments. Parents of the patients were interviewed in their native language using a standardized questionnaire. Biochemical and microbiological tests such as for rubella, reducing sugar and blood glucose were also performed. RESULTS: Out of 172 children, 88.4% had non-traumatic cataract and 11.6% had traumatic cataracts. Among non-traumatic cataracts, 7.2% were hereditary, 4.6% were due to congenital rubella syndrome, 15.1% were secondary and 73.0% were undetermined. In the group of undetermined cases, during pregnancy 67% of the mother had history of illness, and 22% had taken medications during pregnancy. CONCLUSIONS: Our study shows that nearly 12% of non-traumatic cataract is due to potentially preventable causes. Health education of women to childbearing age and school children can decrease incidence of pediatric cataracts.

Comparison of the effect of torsional and microburst longitudinal ultrasound on clear corneal incisions during phacoemulsification.

PURPOSE: To compare incision integrity after clear corneal microcoaxial phacoemulsification using longitudinal and torsional ultrasound (US). SETTING: Iladevi Cataract & IOL Research Centre, Ahmedabad, India. DESIGN: Prospective randomized experimental clinical trial. METHODS: Part 1 comprised an experimental study of rabbit eyes. Group 1 received longitudinal US. Group 2 received torsional US. The right eye of each rabbit served as a control. Samples were processed for histomorphology and collagen I denaturation by immunofluorescence. Part 2 comprised a clinical trial of patients. Group 1 received torsional US. Group 2 received longitudinal US. At the end of surgery, trypan blue 0.0125% was instilled. After 2 minutes, 0.1 mL of aqueous was aspirated and its optical density measured. RESULTS: In part 1, incision histomorphology was comparable in both modalities. Collagen denaturation tests (immunofluorescence, dot blot analysis) showed no irregularity in collagen arrangement in either group. In Group 2, Descemet membrane was detached and endothelial cells were minimal at the roof of the incision. In part 2, trypan blue ingress into the anterior chamber was significantly greater in Group 1 than in Group 2 (mean 3.40 + 0.6 log units versus and 3.77 + 0.82 log units) (P<.007). CONCLUSIONS: Incision histomorphology in the torsional group showed minimal Descemet membrane detachment and minimal endothelial cell loss at the roof of the incision. Minimal ingress of trypan blue into the anterior chamber was observed with torsional US, indicating better wound integrity than with longitudinal US. FINANCIAL DISCLOSURE: No author has a financial or proprietary interest in any material or method mentioned.

Post-operative capsular opacification.

Post-operative capsular opacification (PCO) is a multifactorial physiological consequence of cataract surgery. Opacification involving the central posterior capsule has a significant impact on high and low contrast acuity and low contrast sensitivity. The assessment of PCO on cadaver eyes, experimental studies and culture models and in clinical studies has provided an understanding of its pathogenesis. The proliferation, migration and abnormal differentiation of residual lens epithelial cells and fibers in the capsular bag have been implicated in the pathogenesis of PCO. The incidence and severity of PCO correlates to the use of surgical techniques, intraocular lens (IOL) optic edge designs and IOL materials. This article summarizes the clinical studies with recommendations for retarding the development of central PCO. It discusses experiments with pharmacological agents broadly categorized as anti-inflammatory, immuno-modulating, antiproliferative, antiadhering and antitransdifferentiating agents for the prevention of PCO. These studies will remain critical for future endeavors undertaken for the eradication of PCO.

Post-operative capsular opacification: a review.

Post-operative capsular opacification is a multifactorial physiological consequence of cataract surgery. Opacification involving the central posterior capsule has a significant impact on high and low contrast acuity and low contrast sensitivity. The assessment of Posterior Capsule Opacification (PCO) on cadaver eyes, experimental studies, culture models and in clinical studies has provided an understanding of its pathogenesis. The proliferation, migration and abnormal differentiation of residual lens epithelial cells and fibers in the capsular bag have been implicated in the pathogenesis of PCO. The incidence and severity of PCO correlates to the meticulous use of surgical techniques, IOL optic edge designs and IOL materials. This article summarizes the clinical studies with recommendations for retarding the development of central PCO. It discusses experiments with pharmacological agents broadly categorized as anti-inflammatory, immuno-modulating, antiproliferative, antiadhering, antitransdifferentiating agents for the prevention of PCO. These studies will remain critical for future endeavors undertaken for eradication of PCO.