Towards photo-CIDNP MAS NMR as a generally applicable enhancement method

Solid-state photochemically induced dynamic polarisation (photo-CIDNP) is one of the hyperpolarization techniques that tremendously enhances the sensitivity and selectivity of solid state NMR. Photo-CIDNP MAS NMR studies on entire cells of heliobacteria Hb. mobilis in both anaerobic and aerobic forms are presented in Chapter 2. This chapter demonstrates that photo-CIDNP MAS NMR can be applied as an analytical tool directly on to cells and bacteria even if no further purification is known or possible. To improve the knowledge of the solid-state photo-CIDNP effect, field dependencies are measured over large ranges (Chapter 3). The electron spin density distribution in the 3P donor triplet state is constructed here using the analysis of differential relaxation mechanism and this stresses the versatility of photo-CIDNP MAS NMR as hyperpolarization method. Chapter 4 describes the first observation of photo-CIDNP MAS NM R in phototropin LOV1-C57S, overcoming an old limitation of this method to natural photosynthetic RCs. The first analytical studies on the LOV1 system in Chapter 5 provides the potential to develop photo-CIDNP MAS NMR into an enhancement method generally applicable to electron transfer proteins. In Chapter 6 this leads to a new paradigm, expecting plenty of photo-CIDNP hyperpolarization if the experiment is optimized for the LOV1 system.UBL - phd migration 201

The solid-state photo-CIDNP effect

Solid state NMR/Biophysical Organic Chemistr

The field-dependence of the solid-state photo-CIDNP effect in two states of heliobacterial reaction centers

Solid state NMR/Biophysical Organic Chemistr

Effects of postharvest treatments on storage quality of lime (Citrus latifolia Tanaka) fruit : a thesis presented in partial fulfilment of the requirements for the degree of Doctor of Philosophy in Food Technology at Massey University, New Zealand

Limes (Citrus latifolia Tanaka) are an attractive fruit crop but generally suffer a loss in value as their colour changes from green to yellow. Various approaches were taken to slow degreening including low temperature storage, use of controlled atmosphere (CA) environments, and treatment of fruit with physiologically active agents such as gibberellic acid (GA3). However, the cold storage life of lime fruit can also be restricted by a number of factors including chilling injury (CI) and rots. Various pretreatments such as the use of fungicide (thiabendazole, TBZ) and hot water dipping (HWD) and several postharvest regimes based on temperature conditioning (step down technique) and intermittent warming (IW) regimes were further investigated to protect the fruit against rots and CI during cold storage. The objective of this study was to determine what storage conditions and pretreatments would permit long term storage of NZ limes with minimal loss of quality. CA storage (10% O2 with 0 or 3% CO2) was compared to regular air storage (RA) and IW (varying durations) treatments across a range of temperatures. Although some CA storage regimes could assist in delaying degreening, none of the treatments provided protection against CI. CA storage at 3% CO2 delayed yellowing and gave better fruit quality than the low CO2 treatment. High CO2 CA treatments at 5 or 7°C decreased the rate of colour change compared to other constant temperature treatments but did not protect against CI. CI limited storage of fruit under all conditions at constant low temperatures. Including fungicide (TBZ) in the dip water reduced the incidence of rots and had a secondary effect on protection against CI of lime fruit. However, fungicide use may sometimes exacerbate stresses such as heat injury on lime peel. Hot water dipping has been shown previously to hold potential as a storage pretreatment, but this technique may give risk of damage on produce if it is dipped at too high a temperature. Some HWD treatments did delay degreening, but there was no major effect on CI. HWD at > 47°C for = 4 min caused heat injury to NZ limes. All HWD treatments showed severe CI (>15%) after 10 weeks of cold storage; and HWD fruit stored under RA at 13°C did not show any CI but showed some pitting (= 10%) and degreened rapidly. Overall no suitable HWD treatment for limes was identified in this trial. This project identified the critical periods and temperature conditions for successful IW of limes. The IW conditions successfully delayed losses in quality of lime fruit provided the first warming period was applied within the first 20 days of storage. At least 2-cycle IW was required to maintain lime quality during long term storage. Some benefits were found after just one cycle of IW treatment but there were not enough to extend storage life. IW storage benefited fruit quality and provided the highest overall fruit quality of all postharvest treatments tested. The degreening of lime during cold storage at 5°C could be delayed by IW treatments in which the fruit were stored at 5°C for 12, 16 or 20 days then moved to 15°C for 2 days. Both 2- and 6-cycle IW treatments proved satisfactory for maintaining colour on the green and yellow side of lime for 12 weeks of storage. IW treatments in which fruit were warmed within 20 day of cold storage did not show significant CI symptoms after 12 weeks of storage, and the 2-cycle IW treatment showed only a low percentage of CI fruit at this time. A 2-cycle IW treatment was almost as effective as 6 cycles, and a step down treatment also showed some promising results, indicating that it may be possible to further optimize the time and duration of variable temperature storage regimes to meet both quality requirements and the constraints of temperature management in commercial coolstores. The application of these regimes to other citrus species may also be beneficial. There are a number of physiological explanations that may account for the effectiveness of IW including positive effects on heat shock protein (HSP) and cell membranes. Nutritional factors such as vitamin C and flavonoid compositions were also investigated and fruit that did not show visible CI were found to retain at-harvest levels of these factors. Practical ways of implementing IW are discussed. In order to understand the effectiveness of IW on degreening, I used a logistic model to describe degreening of lime peel. This modelling approach demonstrated that IW did not change the mechanism of lime degreening based on the similarity between the hue values predicted by the model and the actual hue values measured during lime storage. The activation energy (Ea) for degreening based on either hue angle (H°) or colour score (CS) during air storage was estimated to be ~53 and ~86 KJ.mol-1, respectively. Relationship between colour (H° and CS) and chlorophyll content, relationship between reflectance spectra (%), chlorophyll content and H° of lime fruit stored under different conditions are presented and discussed. This data allowed deduction to be made about the changes in individual pigments that are driving colour change during “good” and “bad” storage

Factors Predicting 150 and 200 Microgram Adenosine Requirement during Four Increasing Doses of Intracoronary Adenosine Bolus Fractional Flow Reserve Assessment

Direct intracoronary adenosine bolus is an excellent alternative to intravenous adenosine fractional flow reserve (FFR) measurement. This study, during four increasing adenosine boluses (50, 100, 150, and 200 mcg), aimed to explore clinical and angiographic predictors of coronary stenotic lesions for which the significant ischemic FFR (FFR ≤ 0.8) occurred at 150 and 200 mcg adenosine doses. Data from 1055 coronary lesions that underwent FFR measurement at the Central Chest Institute of Thailand from August 2011 to July 2021 were included. Baseline clinical and angiographic characteristics were analyzed. The FFR ≤ 0.8 occurred at adenosine 150 and 200 mcg boluses in 47 coronary lesions, while the FFR ≤ 0.8 occurred at adenosine 50 and 100 mcg boluses in 186 coronary lesions. After univariable and multivariable logistic regression analyses, four characteristics, including male sex, younger age, non-smoking status, and FFR procedure of RCA, were predictors of the occurrence of FFR ≤ 0.8 at adenosine 150 and 200 mcg doses. Combining all four predictors as a predictive model resulted in an AuROC of 0.72 (95% CI: 0.68–0.76), an 86% negative predictive value. Comparing these four predictors, the FFR procedure of RCA gave the most predictive power, with the AuROC of 0.60 (95% CI: 0.56–0.63)

The Solid-State Photo-CIDNP Effect and Its Analytical Application

Photochemically induced dynamic nuclear polarization (photo-CIDNP) is an effect that produces non-Boltzmann nuclear spin polarization which can be observed as modification of signal intensity in NMR spectroscopy. The effect is well known in liquid-state NMR where it is explained most generally by the classical radical pair mechanism (RPM). In the solid-state, other mechanisms are operative in the spin-dynamics of radical pairs such as three-spin mixing (TSM) and differential decay (DD). Initially the solid-state photo-CIDNP effect has been solely observed on natural photosynthetic reaction centers (RCs). Therefore the analytical capacity of the method has been explored in experiments on reaction centers (RCs) of the purple bacterium of Rhodobacter ( R. ) sphaeroides . Here we will provide an account on phenomenology, theory, and analytical capacity of the solid-state photo-CIDNP effect

The Solid-State Photo-CIDNP Effect and Its Analytical Application

Photochemically induced dynamic nuclear polarization (photo-CIDNP) is an effect that produces non-Boltzmann nuclear spin polarization which can be observed as modification of signal intensity in NMR spectroscopy. The effect is well known in liquid-state NMR where it is explained most generally by the classical radical pair mechanism (RPM). In the solid-state, other mechanisms are operative in the spin-dynamics of radical pairs such as three-spin mixing (TSM) and differential decay (DD). Initially the solid-state photo-CIDNP effect has been solely observed on natural photosynthetic reaction centers (RCs). Therefore the analytical capacity of the method has been explored in experiments on reaction centers (RCs) of the purple bacterium of Rhodobacter ( R. ) sphaeroides . Here we will provide an account on phenomenology, theory, and analytical capacity of the solid-state photo-CIDNP effect

Solid-State Photo-CIDNP Effect Observed in Phototropin LOV1-C57S by 13C Magic-Angle Spinning NMR Spectroscopy

Thamarath SS, Heberle J, Hore PJ, Kottke T, Matysik J. Solid-State Photo-CIDNP Effect Observed in Phototropin LOV1-C57S by 13C Magic-Angle Spinning NMR Spectroscopy. JOURNAL OF THE AMERICAN CHEMICAL SOCIETY. 2010;132(44):15542-15543

Enhancing the sensitivity of micro magnetic resonance relaxometry detection of low parasitemia Plasmodium falciparum in human blood

Upon Plasmodium falciparum infection of the red blood cells (RBCs), the parasite replicates and consumes haemoglobin resulting in the release of free heme which is rapidly converted to hemozoin crystallites. The bulk magnetic susceptibility of infected RBCs (iRBCs) is changed due to ferric (Fe3+) paramagnetic state in hemozoin crystallites which induce a measurable change in spin-spin relaxation (transverse relaxation) rate in proton nuclear magnetic resonance (NMR) of iRBCs. Earlier, our group reported that this transverse relaxation rate (R2) can be measured by an inexpensive, portable 0.5 Tesla bench top magnetic resonance relaxometry (MRR) system with minimum sample preparation and is able to detect very low levels of parasitemia in both blood cultures as well as animal models. However, it was challenging to diagnose malaria in human blood using MRR, mainly due to the inherent variation of R2 values of clinical blood samples, caused by many physiological and genotypic differences not related to the parasite infection. To resolve the problem of baseline R2 rates, we have developed an improved lysis protocol for removing confounding molecular and cellular background for MRR detection. With this new protocol and by processing larger volume of blood (>1 ml), we are able to reliably detect very low level of parasitemia (representing early stage of infection, ~0.0001%) with a stable baseline and improved sensitivity using the current MRR system.NRF (Natl Research Foundation, S’pore)Published versio