eDNA extraction: phenol-chloroform-isoamyl alcohol DNA purification from filters stored in Longmire buffer v1

This is an organic DNA extraction method for filters preserved in 2 ml of Longmire buffer that uses a phase lock to allow easy decanting of the aqueous layer instead of pipetting.

eDNA extraction: phenol-chloroform-isoamyl alcohol DNA purification from filters stored in Longmire buffer v1

This is an organic DNA extraction method for filters preserved in 2 ml of Longmire buffer that uses a phase lock to allow easy decanting of the aqueous layer instead of pipetting.

Safe Blast Design for Efficient and Sustainable Underwater Excavation: Art Meets Science!

Most of the dredging work associated with harbor, port, channel deepening, and other related operation requires underwater blasting due to the characteristics of material being dredged / moved. Underwater blasting is typically used to remove rocks for deepen harbours and channels, creating channels and levees, installing conduits, and other more specialised blasting operations that shall be completed below sea. Usually, such dredging work occurs in deep-water of varying between 16–20m in order to remove just few meters of rocks. Hence, this type of blasting activity needs high level of skill and familiarity than equivalent activities carried out above the surface of water because of aqueous layer over the its rock. Therefore, the factors such as selection of drilling parameters and drilling equipment, selection of appropriate explosives and accessories, usage of correct powder factor, determination of safe explosive charges per delay and selection of suitable personnel are studied carefully for accomplishing the successful underwater blasting operations. In addition to the above, the system shall also address the proper design for the underwater blasts to excavate the rock to the required depth keeping in view the permissible allowances of minimum and maximum depth and fragment size required. While adopting underwater blasts, adequate safety measures are also defined for safety of men, other vessels in the blasting zone and structures from blasting vibrations. Here, the authors broadly outline their approach with respect to underwater blasting using the existing blasting technology, with a case study.

Evaluation of Antioxidant Activity of Caffeine Extracted From Kolanut (Cola Nitida)

Crude caffeine was isolated from kola nut, by treating the filtrate obtained from powdered kola nut with lead (II) acetate to precipitate acids and albumins, along with HCl to precipitate lead ions, and to make some insoluble tannins to be soluble, then Na2CO3 solution was added to neutralise the acidic HCl solution. Dichloromethane is added to the filtrate to extract the caffeine from the aqueous layer and then distilled to obtain the crude caffeine. The extracted crude caffeine is recrystallized with activated charcoal and tested for its antioxidant capacity, using the DPPH assay, which screens the antioxidant activity of plant extracts. The DPPH (2, 2-diphenyl-1-picrylhydrazyl) in methanol helps to facilitate the extraction of the antioxidant compounds from the sample. Ascorbic acid (Vitamin C) is used as the standard for this assay, due to its appreciable antioxidant activity. The IC50 value was calculated from the graph of % inhibition against concentration, and the extracted caffeine from kola nut was found out to have a high antioxidant activity comparable to that of the standard, ascorbic acid.

Environmentally benign PEG-OSO3H catalysed Ultrasound promoted cascade synthesis of furocoumarins in water

Background: Green synthesis of furocoumarins in water, PEG-OSO3H used as a recyclable catalyst for the one-pot syntheis in water. avoid used of toxic, lachrymatric and unstablephenacyl bromides instead of that we prepared it in-situ and directly condence with 4-hydroxycoumarins. Objective: PEG-OSO3H found to be an efficient and reusable catalyst for the microwave irradiated one-pot synthesis of 3- Aryl-furo[3,2-c]coumarins in PEG-water as a greener reaction medium. This method found to be rapid, simple, ecofriendly and yield obtained relatively higher than other reported methods. Methods: Aromatic ketones (5 mmol), NBS (5mmol) was added in solution of PEG-OSO3H (10 mol%) in 10 ml water and irradiated under ultrasound at 80-85°C. then solution of 4-hydroxycoumarin (5 mmol) and solution of acetic acid and ammonium acetate (1 ml) were added into it. Reaction mass was further irradiated for 3 min. The reaction mixture was extracted with ethyl acetate (3 x 10ml) and the combined ethyl acetate phase was concentrated under reduced pressure to obtain corresponding 3-Aryl-furo[3,2-c]coumarins. Aqueous layer was reused for the next reactions. Results: It was observed that electrons withdrawing groups (-NO2,-F etc.) on benzene ring of aromatic ketones increases the rate of reactions as well as yield of the products. The IR spectrum of compound 4a showed prominent peaks at 1737cm-1C=O stretching for lactone, 1H NMR of compound 4c showed characteristic singlets for –CH3 of methoxy group of at 3.94 ppm, The above spectral data confirmed the structure of compound. The 13C NMR spectrum of 4b also shows the peaks of CH3, at 23.25 ppm. Conclusion: PEG-OSO3H found to be an efficient and reusable catalyst for the microwave irradiated one-pot synthesis of 3-Aryl-furo[3,2-c]coumarins in PEG-water as a greener reaction medium. This method found to be rapid, simple, ecofriendly and yield obtained relatively higher than other reported methods.

Recovery and Purification of Glycerine as By-product from Philippine Coconut Methyl Ester

The study is about the recovery and purification of glycerine as a by-product from coconut methyl ester production in the Philippines. The aqueous layer produced from settling or phase splitting of the methyl ester after the transesterification process was subjected to various treatments like acidification, neutralization, concentration in vacuo in order to get back the crude glycerine. The crude glycerine obtained from the laboratory and scale-up process conformed with the specification set by the British standard for crude glycerine. The recovered glycerine is composed of: 84.92%, glycerine; 8.03%, ash; 4.72%, H20; 2.32%, MONG. Further distillation yielded a refined glycerine that meets with the specification set by USP. The average glycerine content of refined glycerine is 96.86%; ash, 0.06%; water, 1.10%, refractive index @ 20°C, 1.4696, specific gravity at 25°C, 1.296 g.

Properties of orb weaving spider glycoprotein glue change during Argiope trifasciata web construction

An orb web’s prey capture thread relies on its glue droplets to retain insects until a spider can subdue them. Each droplet’s viscoelastic glycoprotein adhesive core extends to dissipate the forces of prey struggle as it transfers force to stiffer, support line flagelliform fibers. In large orb webs, switchback capture thread turns are placed at the bottom of the web before a continuous capture spiral progresses from the web’s periphery to its interior. To determine if the properties of capture thread droplets change during web spinning, we characterized droplet and glycoprotein volumes and material properties from the bottom, top, middle, and inner regions of webs. Both droplet and glycoprotein volume decreased during web construction, but there was a progressive increase in the glycoprotein’s Young’s modulus and toughness. Increases in the percentage of droplet aqueous material indicated that these increases in material properties are not due to reduced glycoprotein viscosity resulting from lower droplet hygroscopicity. Instead, they may result from changes in aqueous layer compounds that condition the glycoprotein. A 6-fold difference in glycoprotein toughness and a 70-fold difference in Young’s modulus across a web documents the phenotypic plasticity of this natural adhesive and its potential to inspire new materials.

Structural and Molecular Tear Film Changes in Glaucoma

: The Tear Film (TF) is a trilaminar and dynamic fluid covering the entire Ocular Surface (OS), consisting of a mucus, aqueous, and lipid layer deeply interacting between them. Because of its structure and functions, TF plays a pivotal role in the preservation of the OS integrity and the quality of vision. Medical therapy for glaucoma is recognized to profoundly disturb the OS homeostasis by altering all components of the ocular surface unit, including TF. The presence of preservatives, the number of daily eye drops instillations, and the duration of therapy are the main contributors to TF changes. From the physio-pathological side, TF alterations are induced by toxic and allergic mechanisms and result from goblet cell and Meibomian gland loss, dysfunction of accessory lacrimal glands, and epithelial disruption. : In detail, TF changes are represented by mucus layer thinning, reduced mucin concentration, aqueous layer volume reduction, and lipid layer thinning with increased tear evaporation. Hyper- osmolarity and instability represent the main hallmarks of these changes and are an expression of a iatrogenic form of dry eye. TF undergoes also molecular modifications that primarily reflect a therapy- or disease-induced inflammatory status of the OS. Over the last years, this field of research aimed a progressively growing interest since molecular variations may be considered as potential candidate biomarkers of glaucoma. The aim of this review is to report the main TF changes occurring during glaucoma, exploring the relationship they may have with the glaucoma-related ocular surface disease and the patient quality of life, and their utility as potential biomarkers of disease.

Dynamic assessment of the tear film muco-aqueous and lipid layers using a novel tear film imager (TFI)

PurposeThe objective of the study was to assess a new technology, the tear film imager (TFI), which can dynamically image the muco-aqueous and lipid layers.MethodsProspective pilot case series of individuals with and without dry eye (DE). Two sequential images were obtained with the TFI. Measurements were assessed for reproducibility and compared with clinically derived DE metrics. Individuals were grouped into DE categories based on signs of DE.Results49 patients participated in the study with a mean age of 58.8 years (SD 15.9) and a female majority (69%). Reproducibility of the muco-aqueous layer thickness (MALT) was excellent (r=0.88). MALT measurements significantly correlated with the Schirmer score (r=0.31). Lipid break up time (LBUT) as measured by the TFI significantly correlated with the clinical measure of tear break up time (TBUT) (r=0.73). MALT and LBUT were significantly thinner and shorter, respectively, in the DE groups (mild–moderate and severe) compared with the control group. When comparing TFI parameters to clinically assessed signs, sensitivity of the device was 87% and specificity was 88%.ConclusionThe TFI is the first machine capable of reproducibly measuring muco-aqueous thickness in human subjects which correlates with Schirmer score. In parallel, it assesses other important aspects of tear film function which correlate with clinician assessed DE metrics.