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In embryonic neurons, the intracellular chloride concentration is elevated, making GABA and glycine depolarizing. Later in development, coincident with neuronal maturation, the extruding potassium-chloride co-transporter KCC2 is expressed. It reverses the chloride gradient, rendering it hyperpolarizing. Early depolarization is assumed to play trophic roles during nervous system development. I thus decided to investigate the effects of the depolarizing chloride gradient on development in vivo in the zebrafish embryo. I first determined the temporal pattern of KCC2 expression in zebrafish and found it was absent in the embryo. I then over-expressed wild-type, gain-of-function and loss-of-function variants of human KCC2, using GFP-tagged constructs for detection purposes. Over-expression of functional hKCC2 perturbed the morphology and motor behaviours of the embryos. At the cellular level, KCC2 impaired axonal growth and affected the neuronal populations in the brain, hindbrain and spinal cord. This suggests the depolarizing effects of glycine are critical for neurogenesis.

Although the importance of cation/proton antiporters in cellular physiology is well recognized and widely studied, many antiport systems remain underinvestigated. In this work, I report the phenotypic and biochemical effects of deletion of the cytoplasmic C-terminal tail of the NhaP2 antiporter from Vibrio cholerae (Vc-NhaP2). Namely, deletion of the C-terminal tail results in diminished K+/H+ and Na+/H+ antiport activity, as well as a 5-fold decrease in affinity for its major substrate, K+ (measured as the apparent Km at pH 7.5). Furthermore, reconstitution of antiport activity in the truncation mutant upon addition of exogenous C-terminal tail is demonstrated. Currently, the only known mechanism of antiport is for NhaA, which lacks a cytoplasmic tail. Therefore, these results suggest that NhaP2 may employ a novel mechanism of antiport in which the cytoplasmic tail is directly or indirectly involved.

The normalized difference vegetative index (NDVI) has been correlated with physiological plant parameters and used to evaluate plant growth. There is little information about the use of this technique to detect soybean nutrient deficiencies. The objective of this work was to determine the ability of the NDVI sensor to detect P and K deficiencies, and grain yield reduction, in soybean. During 2010 and 2011, NDVI measurements were made on a soybean field trial site known to exhibit yield responses to both P and K nutrition. Four replicates of 8 levels each of P and K nutrition were evaluated. The NDVI measurements were made with an active proximal sensor held parallel to the soil surface every seven days after V2, and until R2. At each measurement a mean NDVI value was found for each plot. Phosphorus deficiency was detected with the first NDVI measurement. Potassium deficiency was first detected just after V4. Differences in NDVI values due to P or K nutrition increased with continued crop development. There were significant R1 leaf composition and grain yield responses to improved P or K nutrition. The active proximal sensor was able to detect soybean growth differences due to P or K deficiencies in soybean.

A new cryptand, 1,18-bis[(phenylmethoxy)methyl]-3,16,20,23,26,29,32,35-octaoxatetracyclo-[16.9.9,05,10 09,14]hexatriaconta-5,7,9,11,13-pentaene was synthesized from cis-9,19-bis(benzyloxymethyl)-1,4,7,11,14,17-hexaoxacycloeicosane-9,l9-diyldimethanol and l,5-bis(bromomethyl)naphthlene in the presence of t-butoxide base. The overall syntheses involved five steps. The new cryptand compound has a melting point at 164-1650 C, and the yield was 18.5%. The NMR spectra and the crystal structure determination proved the synthesized cryptand the same as that desinged.Fluorescent measurements were performed with this cryptand. The fluorescence was found to be quenched by cesium ion, and the quenching increased with the increasing of cesium ion concentration. The average quenching of the fully complexed (with Cs +) cryptand was estimated as 20 + 2% by a computerized data fitting program. The complexation constant was determined to be 16 ± 5 M-1. A possible explanation for these observations is presented. / Department of Chemistry

Porous constructs of calcium polyphosphate (CPP) are under investigation as a substrate for tissue engineering of cartilage for repair of osteochondral defects. Previous studies have shown that CPP has the required features to satisfy these requirements. However, its degradation rate is lower than desired. This study investigated the effect of doping with MgCO3, MgCl2, K2CO3 or KCl at a molar ratio of M/Ca = 0.02 on sintering and in vitro degradation behavior of CPP. Doping with magnesium or potassium improved the tensile and compressive strengths of CPP at similar porosities. After 15 days of aging in phosphate buffer saline, the rate of tensile strength loss was faster for the doped CPP groups than undoped CPP. The chemical degradation rate of Mg-doped CPP groups was the fastest among CPP groups. The chemical degradation rate of K-doped CPP groups was slower than undoped CPP.

Porous constructs of calcium polyphosphate (CPP) are under investigation as a substrate for tissue engineering of cartilage for repair of osteochondral defects. Previous studies have shown that CPP has the required features to satisfy these requirements. However, its degradation rate is lower than desired. This study investigated the effect of doping with MgCO3, MgCl2, K2CO3 or KCl at a molar ratio of M/Ca = 0.02 on sintering and in vitro degradation behavior of CPP. Doping with magnesium or potassium improved the tensile and compressive strengths of CPP at similar porosities. After 15 days of aging in phosphate buffer saline, the rate of tensile strength loss was faster for the doped CPP groups than undoped CPP. The chemical degradation rate of Mg-doped CPP groups was the fastest among CPP groups. The chemical degradation rate of K-doped CPP groups was slower than undoped CPP.

The rapid accumulation of municipal solid waste is a significant environmental concern in our rapidly growing world. Due to its low cost, high energy recovery and limited environmental impact anaerobic digestion (AD) is a promising solution for stabilizing the organic fraction of municipal solid waste (OFMSW). Hydrolysis is often the rate-limiting step during AD of wastes with high solid content; this step can be accelerated by pretreatment of waste prior to AD. This thesis presents the results of alkaline pretreatment of OFMSW using NaOH and KOH. Four different pH levels 10, 11, 12 and 13 at two temperatures 23±1°C and 80±1°C were examined to study the effects of the pretreatment on (i) enhancing the solubility of the organic fraction of the waste, and (ii) enhancing the AD process and the biogas production. The effects on solubility were investigated by measuring changes in the soluble COD (SCOD) concentrations of pretreated wastes and the enhanced AD was investigated by measuring volatile solids (VS) destruction, total COD (TCOD) and SCOD removal in addition to biogas and methane production using biochemical methane potential (BMP) assay and semi-continuous laboratory reactor experiments. Pretreatment at pH 13 at 80±1°C demonstrated the maximum solubility for both NaOH and KOH pretreated samples; however the BMP analysis demonstrated that pretreatment at pH 12 at 23±1°C showed the greatest biogas yield relative to the removed VS for both chemicals. Thus pretreatment at pH 12 at 23±1°C using NaOH and KOH were examined using semi-continuous reactors at three different HRTs: 10, 15 and 20 days. Pretreatment demonstrated a significant improvement in the AD performance at SRTs of 10 and 15 days.

Two series of tests were performed in summer 1993 and repeated in summer 1994 to investigate effects of potassium, calcium and water table depth on mechanical properties of tomato fruit. In the first group of tests five mechanical properties stress, strain, ER (ratio of stress to strain at break), energy and toughness were calculated from the force-deformation curve obtained by compression of pericarp disks taken from tomato fruits (cv New Yorker) at the breaker stage. In the second group of tests force, deformation and energy were obtained from puncture tests on whole tomato fruit. The textural parameters were used to compare the effects of the potassium, calcium and water level depths on the mechanical properties of tomatoes. Results showed the effects of calcium and water table depths on the mechanical properties were significant. Tomato fruits exposed to increasing moisture stress levels by increasing water table depth exhibited increased firmness. Increased application of calcium also resulted in increased firmness. Application of different potassium levels had in general no significant effect on breaking stress, strain, ER, energy and toughness of tomato fruit pericarp. / A new method of measuring skin strength of tomatoes is described. The method, refered to as loop method, overcame the problems with the existing method using mechanical grips. Further, the loop method was applied to measure the effect of potassium and calcium on skin strength of tomato. Using the loop method, data were collected on relaxation of tomato skin (epidermis). A computer program was written to model th tensile stress relaxation behaviour of tomato skin. The program was employed to calculate linear and non-linear coefficients of a relaxation model. The fitted values were in a very good agreement with experimental data (R$ sp2>$0.99).

Typescript. / Thesis (Ph. D.)--University of Hawaii, 1966. / Bibliography: leaves 156-168. / xiii, 168 l illus. (part mounted), tables