Effect of algal inoculation on the yield and vitamin C content of two varieties of tomato

Summary Two varieties of tomato (Pusa Rubi and Selection 120) positively responded to algal inoculation in terms of the yield of fruits and shoots, but there was no significant effect on the vitamin C content of the fruits. A combined application of urea and algae was more effective than the application of urea alone.     

A Study of Maternal and Maternal Interaction Effects in Diallel Crosses

The analysis of diallel crosses by including the components due to maternal effect and maternal interaction effects have been presented for Griffing method—1 (random effect model) and Griffing method—3 (fixed and random effect model). Wherever exact test of significance is not possible, testing procedure using Satterthwaite (1946) approximation has been presented.     

Papain Does Not Cleave Operator-Bound Lambda Repressor: Structural Characterization of the Carboxy Terminal Domain and the Hinge

Abstract

The circular dichroism spectra of three different purified carboxy terminal fragments 93–236, 112–236 and 132–236 of the bacteriophage γ cI repressor have been measured and compared with those of the intact repressor and the amino terminal fragment 1–92. All three carboxy terminal fragments contain mostly β-strands and loops, a minor helix content increasing with the size of the fragment, showing that the 93–131 region previously called a hinge is structured. Fourier transformed infrared spectra also showed that fragment 93–236 contains α-helices, β-sheets and turns but fragment 132–236 contains no detectable α-helix, only β-sheets and turns. Papain is known to cleave the γ repressor, but it is shown here that it cannot cleave the operator-bound repressor dimer. For the 132–236 fragment, both the wt and the SN228 mutant previously shown to be dimerization defective in the intact, gave similar dimerization properties as investigated by HPLC at 2 to 100 µM protein concentration, with a KD of 13.2 µM and 19.1 µM respectively. The papain cleavage for wt and SN228 proceed at equal rates for the first cleavage at 92–93; however, the subsequent cleavages are faster for SN228. The three Cys residues in the 132–236 fragment were found to be unreactive upon incubation with DTNB, indicating the thiol sulfur atoms are buried in the repressor carboxy terminal domain. Denaturation of the 132–236 fragment studied by tryptophan fluorescence shows two transitions centered at 1.5 M and 4.5 M of urea.     

Quantifying Single Microvessel Permeability in Isolated Blood-perfused Rat Lung Preparation

The isolated blood-perfused lung preparation is widely used to visualize and define signaling in single microvessels. By coupling this preparation with real time imaging, it becomes feasible to determine permeability changes in individual pulmonary microvessels. Herein we describe steps to isolate rat lungs and perfuse them with autologous blood. Then, we outline steps to infuse fluorophores or agents via a microcatheter into a small lung region. Using these procedures described, we determined permeability increases in rat lung microvessels in response to infusions of bacterial lipopolysaccharide. The data revealed that lipopolysaccharide increased fluid leak across both venular and capillary microvessel segments. Thus, this method makes it possible to compare permeability responses among vascular segments and thus, define any heterogeneity in the response. While commonly used methods to define lung permeability require postprocessing of lung tissue samples, the use of real time imaging obviates this requirement as evident from the present method. Thus, the isolated lung preparation combined with real time imaging offers several advantages over traditional methods to determine lung microvascular permeability, yet is a straightforward method to develop and implement.     

NMR localization of the O-mycoloylation on PorH,a channel forming peptide from Corynebacterium glutamicum

PorH and PorA are two small peptides that, in complex, form a voltage-dependent ion channel in the outer membrane of Corynebacterium glutamicum. Specific post-translational modifications on PorA and PorH are required for the formation of a functional ion channel. The assignment of PorH proton NMR chemical shifts in DMSO, allowed identifying unambiguously the exact position of the PorH O-mycoloylation on Ser 56 side chain. This was further confirmed by site directed mutagenesis and mass spectrometry. Together with the previously published localization of PorA mycoloylation, this provides the complete primary structure characterization of this outer membrane porin.     

Purification, characterization and USAGE of thermotolerant laccase FROM Bacillus sp. FOR biodegradation of synthetic dyes

A Bacillus sp. isolated from sediments of distillery unit was found to overproduce laccase when cultured in a synthetic media containing 1mM CuSO₄ and 10% distillery spent wash as inducers along with 1% dextrose (w/v) and 0.1% tryptone (w/v) as additional carbon and nitrogen sources. The extracellular purified enzyme was highly thermostable with a calculated half-life of 23 min at 75°C. The optimal pH and temperature of the Bacillus sp. laccase were recorded to be 3.0 and 35°C, respectively. Sodium azide and solvents like methanol and acetonitrile completely inhibited enzyme activity. The average molecular weight of the purified enzyme as determined by SDS-PAGE and zymogam studies was around 70 kDa. Kinetic parameters were detected by using 2,2′-azinobis-(3-ethylbenzthiazoline-6-sulfonate) (ABTS) as substrate. At high ABTS concentrations (> 6 mM) a substrate inhibition phenomenon appeared and K _M (0.60 mM), V _max (983.00 U/min) values were determined. The polypeptide sequences showed significant similarity with Cudependent oxidoreductases through MALDI-TOF MS analysis. In addition, the crude Bacillus sp. laccase showed enormous potential for decolorization of various recalcitrant dyes. The apparent high stability of this enzyme makes it a good candidate for its possible application in biotechnology.     

Electrochemical immunosensor for label free epidermal growth factor receptor (EGFR) detection

This paper presents an electrochemical immune sensor for label free detection of epidermal growth factor receptor (EGFR) by immobilizing anti-EGFR antibody (Anti-EGFRab) on dithiobissuccinimidyl propionate (DTSP) self-assembled monolayer (SAM) on gold (Au) electrode. Electrochemical studies show that increased surface concentration of redox moieties onto Anti-EGFRab/DTSP immuno-electrode leads to high electron transport and improved sensing performance. The antigen-antibody complex demonstrates a high association constant (5×10(12)L/mol) that results in high affinity of Anti-EGFRab to EGFR, confirming that the DTSP-SAM provides a conducive environment for anti-EGFR immobilization. The electrochemical response of EA/Anti-EGFRab/DTSP/Au electrode as a function of EGFR concentrations exhibits a linear range from 1pg/mL to 100ng/mL, a detection limit of 1pg/mL at a sensitivity of 2.02μAM(-1)at a regression coefficient of 0.99.     

Inner Nuclear Layer Thickening Is Inversley Proportional to Retinal Ganglion Cell Loss in Optic Neuritis

Aim

To examine the relationship between retinal ganglion cell loss and changes in the inner nuclear layer (INL) in optic neuritis (ON).

Methods

36 multiple sclerosis (MS) patients with a history of ON and 36 age and sex-matched controls underwent Optical Coherence Tomography. The paramacular retinal nerve fiber layer (RNFL), combined ganglion cell and inner plexiform layers (GCL/IPL) and inner nuclear layer (INL) thickness were measured at 36 points around the fovea. To remove inter-subject variability, the difference in thickness of each layer between the ON and fellow eye of each patient was calculated. A topographic analysis was conducted.

Results

The INL of the ON patients was thicker than the controls (42.9µm versus 39.6µm, p=0.002). ON patients also had a thinner RNFL (27.8µm versus 32.2µm, p<0.001) and GCL/IPL (69.3µm versus 98.1µm, p<0.001). Among the controls, there was no correlation between RNFL and GCL/IPL as well as RNFL and INL, but a positive correlation was seen between GCL/IPL and INL (r=0.65, p<0.001). In the ON group, there was a positive correlation between RNFL and GCL/IPL (r=0.80, p<0.001) but a negative correlation between RNFL and INL (r=-0.61, p<0.001) as well as GCL/IPL and INL (r=-0.44, p=0.007). The negative correlation between GCL/IPL and INL strengthened in the ON group when inter-subject variability was removed (r=-0.75, p<0.001). Microcysts within the INL were present in 5 ON patients, mainly in the superior and infero-nasal paramacular regions. While patients with microcysts lay at the far end of the correlation curve between GCL/IPL and INL (i.e. larger INL and smaller GCL/IPL compared to other patients), their exclusion did not affect the correlation (r= -0.76, p<0.001).

Conclusions

INL enlargement in MS-related ON is associated with the severity of GCL loss. This is a continuous relationship and patients with INL microcysts may represent the extreme end of the scale.