Cereal protoplast recalcitrance

Summary Cereal leaf protoplasts are extremely difficult to culture (recalcitrant) in vitro. There have been few reports of division and the protoplasts typically exhibit excessive enlargement and vacuolization with reduced cell wall deposition. Inasmuch as leaf base explants are capable of callus formation in vitro, protoplasts derived from this tissue must have lost the ability to divide as a consequence of changes induced by the wall-digestion process. We review evidence suggesting that the inhibition of mitosis in these protoplasts is a consequence of a cascade of events initiated at the plasma membrane. The enzyme treatment necessary for wall removal triggers membrane depolarization and other changes that can lead to the initiation of lipid peroxidation and oxidative stress. Mitotically inactive cereal leaf protoplasts are unable to mount a protective response to these degradative processes. Consequently, the resulting membrane perturbations and permeabilization give rise to secondary effects on the cytoskeleton and the cell wall. These effects include reduced or absent microtubules as well as reduced and uneven wall deposition. Such abnormalities are observed in cereal leaf protoplasts and are sufficient to account for recalcitrance because the occurrence of mitosis is strongly dependent on a normal cell wall and cytoskeleton. This paper is NRCC number 32475.     

Benzothiazole analogs as potential anti-TB agents: computational input and molecular dynamics

Biotin is very important for the survival of Mycobacterium tuberculosis. 7,8-Diamino pelargonic acid aminotransaminase (DAPA) is a transaminase enzyme involved in the biosynthesis of biotin. The benzothiazole title compounds were investigated for their in vitro anti-tubercular activity against two tubercular strains: H37Rv (ATCC 25,177) and MDR-MTB (multidrug-resistant M. tuberculosis, resistant to isoniazid, rifampicin, and ethambutol) by an agar incorporation method. The possible binding mode and predicted affinity were computed using a molecular docking study. Among the synthesized compounds in the series, the title compound {2-(benzo[d]thiazol-2-yl-methoxy)-5-fluorophenyl}-(4-chlorophenyl)-methanone was found to exhibit significant activity with minimum inhibitory concentrations of 1 μg/mL and 2 μg/mL against H37Rv and MDR-MTB, respectively; this compound showed the highest binding affinity (–24.75 kcal/mol) as well.     

Synthesis and urease inhibitory potential of benzophenone sulfonamide hybrid in vitro and in silico

This study deals with the synthesis of benzophenone sulfonamides hybrids (1–31) and screening against urease enzyme in vitro. Studies showed that several synthetic compounds were found to have good urease enzyme inhibitory activity. Compounds 1 (N′-((4′-hydroxyphenyl)(phenyl)methylene)-4′′-nitrobenzenesulfonohydrazide), 2 (N′-((4′-hydroxyphenyl)(phenyl)methylene)-3′′-nitrobenzenesulfonohydrazide), 3 (N′-((4′-hydroxyphenyl)(phenyl)methylene)-4′′-methoxybenzenesulfonohydrazide), 4 (3′′,5′′-dichloro-2′′-hydroxy-N′-((4′-hydroxyphenyl)(phenyl)methylene)benzenesulfonohydrazide), 6 (2′′,4′′-dichloro-N′-((4′-hydroxyphenyl)(phenyl)methylene)benzenesulfonohydrazide), 8 (5-(dimethylamino)-N′-((4-hydroxyphenyl)(phenyl)methylene)naphthalene-1-sulfono hydrazide), 10 (2′′-chloro-N′-((4′-hydroxyphenyl)(phenyl)methylene)benzenesulfonohydrazide), 12 (N′-((4′-hydroxyphenyl)(phenyl)methylene)benzenesulfonohydrazide) have found to be potently active having an IC₅₀ value in the range of 3.90–17.99?µM. These compounds showed superior activity than standard acetohydroxamic acid (IC₅₀?=?29.20?±?1.01?µM). Moreover, in silico studies on most active compounds were also performed to understand the binding interaction of most active compounds with active sites of urease enzyme. Structures of all the synthetic compounds were elucidated by ¹H NMR, ¹³C NMR, EI-MS and FAB-MS spectroscopic techniques.     

Environmental and economic impact of using new-generation wide-base tires

Purpose

New-generation wide-base tire (NG-WBT) is known for improving fuel economy and at the same time for potentially causing a greater damage to pavement. No study has been conducted to evaluate the net environmental saving of the combined system of pavements and NG-WBT. This study adopted a holistic approach (life cycle assessment [LCA] and life cycle costing [LCC]) to quantitatively evaluate the environmental and economic impact of using NG-WBT.

Methods

The net effect of different levels of market penetration of NG-WBT on energy consumption, global warming potential (GWP), and cost based on the fatigue cracking and rutting performance of two different asphalt concrete (AC) pavement structures was evaluated. The performance of pavements was determined based on pavement design lives; pavement surface characteristics, and pavement critical strain responses obtained from the artificial neural network (ANN) based on finite element (FE) simulations were used to calculate design lives of pavements. Based on the calculated design lives, life cycle inventory (LCI) and cost databases, and rolling resistance (RR) models previously developed by the University of Illinois at Urbana-Champaign (UIUC) were used to calculate the environmental and economic impact of the combined system.

Results and discussion

The fuel economy improvement using NG-WBT is 1.5% per axle. Scenario-based case studies were conducted. Considering 0% NG-WBT market penetration (or 100% standard dual tire assembly [DTA]) as a baseline, scenario 1 assumed the same fatigue and rutting potential between NG-WBT and DTA; therefore, the only difference came from fuel economy improvement of using NG-WBT. In scenario 2, pavement fatigue cracking potential determined the pavement design life; both thick and thin AC overlay sections experienced positive net environmental savings, but mixed net economic savings. In scenario 3, pavement rutting potential determined the pavement design life; the thick AC overlay section experienced positive net environmental savings, but mixed net economic savings. The thin section experienced negative net environmental and economic savings.

Conclusions

The outcomes of scenario-based case studies indicated that NG-WBT can result in significant savings in life cycle energy consumption and cost, and GWP; however, these benefits were sensitive to the method used to determine the pavement performance; especially, a small change in pavement strain can result in significant change in pavement life. In addition, the effect of fuel price/economy improvement, discount rate, and International Roughness Index (IRI) threshold values was studied in the sensitivity analyses.

     

Rapid tilt-series acquisition for electron cryotomography

Using a new Titan Krios stage equipped with a single-axis holder, we developed two methods to accelerate the collection of tilt-series. We demonstrate a continuous-tilting method that can record a tilt-series in seconds, but with loss of details finer than ～4?nm. We also demonstrate a fast-incremental method that can record a tilt-series several-fold faster than current methods and with similar resolution. We characterize the utility of both methods in real biological electron cryotomography workflows. We identify opportunities for further improvements in hardware and software and speculate on the impact such advances could have on structural biology.     

Combination of arsenic trioxide and cisplatin synergistically inhibits both hexokinase activity and viability of Ehrlich ascites carcinoma cells

Hexokinase‐2 is overexpressed in several carcinomas including breast cancer to sustain energy for rapidly dividing cells and associates with chemoresistance. However, the impact of chemo drugs (alone or in combination) on hexokinase activity and autophagic cell death is unclear. In this report, we used an in vivo murine adenocarcinoma model to validate the effects of As₂O₃ and cisplatin on hexokinase activity and autophagic cancer cell death. We found that the two drugs inhibit hexokinase activity and induce autophagic marker, beclin 1 expression. Interestingly, combining As₂O₃ with cisplatin synergistically enhanced these effects and alleviated oxidative stress often encountered in As₂O₃ treatment. Altogether, our data provide direct evidence that inhibition of hexokinase activity and induction of autophagic cell death are mediating the antineoplastic effects of As₂O₃ and cisplatin. Our findings raise the potential of combining As₂O₃ with cisplatin as an approach to augment cisplatin‐induced cell death and combat cisplatin chemoresistance in cancer.     

Brain slice stimulation using a microfluidic network and standard perfusion chamber

We have demonstrated the fabrication of a two-level microfluidic device that can be easily integrated with existing electrophysiology setups. The two-level microfluidic device is fabricated using a two-step standard negative resist lithography process. The first level contains microchannels with inlet and outlet ports at each end. The second level contains microscale circular holes located midway of the channel length and centered along with channel width. Passive pumping method is used to pump fluids from the inlet port to the outlet port. The microfluidic device is integrated with off-the-shelf perfusion chambers and allows seamless integration with the electrophysiology setup. The fluids introduced at the inlet ports flow through the microchannels towards the outlet ports and also escape through the circular openings located on top of the microchannels into the bath of the perfusion. Thus the bottom surface of the brain slice placed in the perfusion chamber bath and above the microfluidic device can be exposed with different neurotransmitters. The microscale thickness of the microfluidic device and the transparent nature of the materials [glass coverslip and PDMS (polydimethylsiloxane)] used to make the microfluidic device allow microscopy of the brain slice. The microfluidic device allows modulation (both spatial and temporal) of the chemical stimuli introduced to the brain slice microenvironments.