A method for DNA extraction and molecular identification of Aphids.

Aphid species (Insecta, Hemiptera) are economically important invasive pest throughout the world, though their identification is intricate due to tiny size and inconspicuous nature of morphology. Mitochondrial cytochrome c oxidase I (mtCOI) region has been proven to be a standard barcode to identify the diverse array of insect groups. Isolation of good quality DNA is a fundamental first step in insect DNA barcoding which is obtained by standardizing the DNA isolation method. In this study, we demonstrate a modified CTAB method for the isolation of DNA to maximize the quality and yield from small aphids. This method will help the researchers to efficiently isolate DNA from small aphid and the method can be utilized for other small insects as well. We evaluated the quality of the isolated DNA and the mtCOI gene region were subjected to PCR amplification. Further, the gene segment was sequenced and gene annotation was done by NCBI BLAST program through which the insect was found to be Aphis gossypii. This study provides a set of molecular tools that can be used for identification of insect at species level through DNA barcoding and biodiversity analysis.•Detailed method to maximize quality and quantity of genomic DNA isolated from aphids.•Molecular identification of aphids using mtCOI gene amplification and sequence validation.•First report on Aphis gossypii infecting Solanum trilobatum provides insights of pest identification and management.

Ultrasonically driven green synthesis of palladium nanoparticles by Coleus amboinicus for catalytic reduction and Suzuki-Miyaura reaction.

A novel ultrasonically driven bio-reduction method was adopted to reduce the palladium chloride into palladium nanoparticles (PdNPs@CA) using coleus amboinicus extract as a green synthetic protocol. XRD confirms the formation of phase pure cubic Pd nanoparticles with the crystallite size range of 40-50 nm. The UV-vis spectrum reveals the formation of Pd nanoparticles by the disappeared peak at 480 nm of PdCl2 solution. The size distribution and surface morphology of prepared Pd nanoparticles showed spherical shaped nanoparticles with less agglomeration. The catalytic reduction behaviour of the Pd suspension is studied by 4-nitro phenol reduction process in 8 min further confirms its high catalytic performance. Synthesized PdNPs@CA were explored in ultrasound promoted Suzuki-Miyaura coupling reaction to determine the catalytic behaviour with ultrasonic frequency of 40 kHz and power of 150 W (Power sonic 410 bath sonicator) and its recycling ability is determined. It was found that aryl halides reacted with aryl boronic acids to obtain biaryl compound with excellent reaction yields in the presence of PdNPs@CA only in 30 min using PEG-400 as a green solvent. PdNPs@CA can be recovered efficiently and reused for 7 cycles without loss of its catalytic property.

Comparison of continuous cardiac output monitoring derived from regional impedance cardiography with continuous thermodilution technique in cardiac surgical patients.

Background: Cardiac output (CO) assessment is a corner stone in advanced haemodynamic management, especially in critical ill patients. The present study was conducted to validate cardiac index and cardiac output by NICaS™ with the thermodilution technique using pulmonary artery catheter in post-operative cardiac surgical patients. Materials and Methods: This was a prospective observational clinical study conducted at a tertiary care hospital. 23 adult patients in the age range of 18-65 years who had undergone for elective coronary artery bypass grafting were included in the study. Results: Spearman's correlation coefficient of cardiac index between continuous Thermodilution (cTD) and Non-Invasive Cardiac System (NICaS™) showed a good correlation (r = 0.765, 95% confidence interval 0.70 to 0.82, P < 0.0001). There was a good correlation between cTD and NICaS™ for cardiac output (r = 0.759, 95% confidence interval 0.69 to 0.81, P < 0.0001), Bland-Altman plot for cardiac index between cTD and NICaS™ showed a mean bias of -0.66 ± 0.6919 with limits of agreement being -2.02 to 0.6936. Bland-Altman plot for cardiac output between cTD and NICaS™ showed a mean bias of -1.0386 ± 1.17 with limits of agreement being -3.34 to + 1.26. Percentage error for cardiac index and cardiac output were 64.78% and 64% respectively. Polar plot analysis showed an angular bias of 6.32° with radial limits of agreement being -8.114° to 20.75° for cardiac index and angular bias of 5.6682° with radial limits of agreement being -9.1422° to 20.4784° for cardiac output. Conclusion: NICaS™ demonstrated a good trending ability for both CI and CO. However, NICaS™ derived parameters are not interchangeable with the values derived from continuous thermodilution technique.

Safety of emamectin benzoate administered in feed to Nile tilapia Oreochromis niloticus (L.).

Emamectin benzoate (EB) premix top-coated onto feed is extensively used to treat ectoparasitic crustacean infestations in aquaculture. This study evaluated the safety of EB-dosing in Nile tilapia Oreochromis niloticus at the recommended dose and dosage of 50 µg/kg biomass/day for 7 consecutive days (1X) and compared with control and 10 times the recommended dose (10X). Depletion of EB-residues in the edible muscle of 1X-dosed Nile tilapia was also studied. Mortality, behavioural changes, feed consumption, biomass, EB-residue depletion, and histopathological alterations in the kidney, liver and intestine were determined at slated intervals. Significant dose-dependent reduction in feed intake and biomass and insignificant mortalities were noted in 1X and 10X EB-dosed fish. In 1X EB-dosed fish muscle, the residues peaked on day 7 EB-dosing (9.72 ng/g) and decreased subsequently. Nevertheless, the residue levels were within the acceptable limit of the European Commission and the Canadian Food Inspection Agency even during the EB-dosing period. Histologically, tubule degeneration in the kidney, mild glycogen vacuolation in the liver, and loss of absorptive vacuoles, inflammation and disintegration of the epithelial layer in the intestine of Nile tilapia fed the 1X EB-diet were observed. The fish reverted back to their normal functions with time upon termination of oral-EB-dosing. This work contributed scientific data on the safety of EB particularly on the feed intake, growth reduction, mortality, histopathological alterations, and EB-residue levels in the edible tissues of Nile tilapia fed at the recommended dose and dosage, which suggested that EB-therapy might be reasonably risky in a tropical climate.

Screening Natural Content of Water-Soluble B Vitamins in Fish: Enzymatic Extraction, HILIC Separation, and Tandem Mass Spectrometric Determination.

Despite the potential of LC with tandem MS (MS/MS) in improving sensitivity and selectivity, analytical methods are scarce for the determination of protein-bound and phosphorylated forms of B vitamins in food. This prompted us to develop a method for LC-MS/MS determination of naturally occurring nicotinamide, nicotinic acid, thiamine, pyridoxine, riboflavin, pantothenic acid, biotin, folic acid, and cyanocobalamin in fish. Baseline separation of the vitamins was achieved in a hydrophilic interaction LC condition. An ultrasonication-assisted enzymatic extraction protocol for sample preparation was optimized and validated. The time required for extraction was significantly reduced (to 4 h), while maintaining good extraction efficiency. Acetonitrile content (80%, v/v) in the prepared sample was found to be optimum for excellent peak shape and sensitivity. The dynamic linear range of the vitamins ranged from 2.5 to 500 ng/g, and the regression coefficient values were greater than 0.99. LOQ values ranged from 0.4 to 50 ng/g for the different vitamins. The spike recovery values at 50 and 100 ng/g ranged from 87.5 to 97.5%. The intra- and interday precision values were satisfactory. Accuracy of the developed method was determined by analysis of a Certified Reference Material. The method could also be used for unambiguous determination of the natural content of the target vitamins in fish.

Development of thiamine and pyridoxine loaded ferulic acid-grafted chitosan microspheres for dietary supplementation.

Therapeutic potential of water soluble vitamins has been known for long and in recent times they are being widely supplemented in processed food. Phenolic acid-grafted chitosan derivatives can serve as excellent biofunctional encapsulating materials for these vitamins. As a proof of concept, thiamine and pyridoxine loaded ferulic acid-grafted chitosan microspheres were developed. Ferulic acid was successfully grafted on chitosan by a free radical mediated reaction and the structure was confirmed by FTIR and NMR analysis. When compared to FTIR spectra of chitosan, intensity of amide I (at around 1644 cm(-1)) and amide II (at around 1549 cm(-1)) bands in spectra of ferulic acid-grafted chitosan were found increased, indicating formation of new amide linkage. Strong signals at δ = 6.3-7.9 ppm corresponding to methine protons of ferulic acid were observed in NMR spectra of ferulic acid-grafted chitosan, suggesting the successful grafting of ferulic acid onto chitosan. Grafting ratio of the derivative was 263 mg ferulic acid equivalent/g polymer. Positively charged particles (zeta potential 31 mv) of mean diameter 4.5 and 4.8 µ, corresponding to number distribution and area distribution respectively were observed. Compact microspheres with smooth surfaces and no apparent cracks or pores were observed under scanning electron microscope. Efficient microencapsulation was further proved by X-ray diffraction patterns and thermal analysis. Preliminary anti-inflammatory activity of the vitamin-loaded microspheres was demonstrated.

Transesophageal echocardiography estimation of coronary sinus blood flow for the adequacy of revascularization in patients undergoing off-pump coronary artery bypass graft.

AIMS AND OBJECTIVES: Physiologically coronary sinus (CS) drains the left coronary artery (LCA) territory. Stenosis of the branches of LCA may decrease the coronary sinus blood flow (CSBF). Any intervention that aims at restoring the flow of the stenosed vessel increases coronary artery flow that should consequently increase the CSBF. Hence, this study was undertaken to assess the CSBF before and after each branch of LCA to determine the adequacy of surgical revascularization in patients undergoing elective off pump coronary artery bypass grafting (OPCAB) using transesophageal echocardiography (TEE). MATERIALS AND METHODS: Thirty consecutive patients scheduled for elective OPCAB were enrolled. CSBF was assessed before and after each branch of LCA revascularization using TEE. Left internal mammary artery (LIMA) Doppler was also obtained post LIMA to left anterior descending (LAD) grafting. RESULTS: Hemodynamic and echocardiographic variables were compared by means of Student's t-test for paired data before and after revascularization. The CSBF per beat (1.28 ± 0.71), CSBF per minute (92.59 ± 59.32) and total velocity time integral (VTI) (8.93 ± 4.29) before LAD grafting showed statistically significant increase to CSBF per beat (1.70 ± 0.89), CSBF per minute (130.72 ± 74.22) and total VTI (11.96 ± 5.68) after LAD revascularization. The CSBF per beat (1.67 ± 1.03), CSBF per minute (131.91 ± 86.59) and total VTI (11.00 ± 5.53) before obtuse marginal (OM) grafting showed statistically significant increase to CSBF per beat (1.91 ± 1.03), CSBF per min (155.20 ± 88.70) and total VTI (12.09 ± 5.43) after OM revascularization. In 9 patients, color flow Doppler of LIMA could be demonstrated which showed diastolic predominant blood flow after LIMA to LAD grafting. CONCLUSION: Demonstration of CSBF was simple and monitoring the trend of CSBF values before and after each graft of LCA territory will guide to determine the adequacy of surgical revascularization.

Changes in tissue defence system in white spot syndrome virus (WSSV) infected Penaeus monodon.

The present study examined the changes occurring in the pro phenoloxidase system and antioxidant defence status in haemolymph, hepatopancreas and muscle tissue of white spot syndrome virus (WSSV) infected Penaeus monodon. Tiger shrimps (P. monodon) were infected with white spot virus by intramuscular injection of the virus inoculum. Levels of lipid peroxides and the activities of phenoloxidase, glutathione-dependent antioxidant enzymes [glutathione peroxidase (GPX), glutathione-S-transferase (GST)] and antiperoxidative enzymes [superoxide dismutase (SOD) and catalase (CAT)] were determined. WSSV infection induced a significant increase in lipid peroxidation in haemolymph, muscle and hepatopancreas of experimental P. monodon compared to normal controls. This was paralleled by significant reduction in the activities of phenol oxidase, glutathione-dependent antioxidant enzymes and antiperoxidative enzymes. The results of the present study indicate that the tissue antioxidant defence system in WSSV infected P. monodon is operating at a lower rate, which ultimately resulted in the failure of counteraction of free radicals, leading to oxidative stress as evidenced by the increased level of lipid peroxidation.

Multiple sclerosis in childhood: presenting as bilateral optic neuritis.

Multiple sclerosis is a disease of adolescence and middle age. Though rare, it is being increasingly recognized in younger population. We describe two patients in whom multiple sclerosis was diagnosed before 10 years of age. Both patients had bilateral optic neuritis during the course of their illness with partial recovery; other lesions in the neuraxis recovered reasonably well.