Functional defect and restoration of temperature-sensitive mutants of FlhA, a subunit of the flagellar protein export apparatus

The flagellar axial component proteins are exported to the distal end of the growing flagellum for self-assembly by the flagellar type III export apparatus. FlhA is a key membrane protein of the export apparatus, and its C-terminal cytoplasmic domain (FlhA_C) is a part of an assembly platform for the three soluble export components, FliH, FliI, and FliJ, as well as export substrates and chaperone–substrate complexes. FlhA_C is composed of a flexible linker region and four compact domains (A_CD1–A_CD4). At 42 °C, a temperature-sensitive (TS) G368C mutation in FlhA_C blocks the export process after the FliH–FliI–FliJ–substrate complex binds to the assembly platform, but it remains unknown how it does so. In this study, we analyzed a TS mutant variant, FlhA_C(G368C), and its pseudorevertant variants FlhA_C(G368C/L359F), FlhA_C(G368C/G364R), FlhA_C(G368C/R370S), and FlhA_C(G368C/P550S) using far-ultraviolet circular dichroism. Whereas the denaturation of the wild-type FlhA_C occurs in a single step, FlhA_C(G368C) and its pseudorevertant variants showed thermal transitions, at least, in two steps. The first transition of FlhA_C(G368C) can further be divided into reversible and following irreversible transitions, which correspond to the denaturation of A_CD2 and A_CD1, respectively. We show the relation between the reversible transition and the TS defect in the exporting function of FlhA_C(G368C) and that the loss of function is caused by denaturation of A_CD2. We suggest that A_CD2 is directly involved in the translocation of export substrates.     

Surface receptors and immune activity of purified human circulating mononuclear cells: IV. The demonstration of seven subclasses of T cells in the circulation of the normal individual: The cytotoxic activities of these cells

T lymphocytes were isolated from monocyte-depleted mononuclear cells of normal individuals by rosetting them with sheep erythrocytes. These purified T cells were preferentially depleted of cells with receptors for FcG (T_G cells), FcM (T_M cells), or C3 (T_C cells) by rosette formation with EA(G), EA(M), and EAC, respectively, before or after incubation for 24 hr in medium 199 fortified with fetal calf serum (20%). The unfractionated lymphocytes and the purified and the depleted T cells were analyzed for receptors to FcG, FcM, and C3 and for cytotoxic activity in the natural killer (NK), antibody dependent cell-mediated cytotoxicity (ADCC), and mitogen-induced cell-mediated cytotoxicity (MICC) assays. The T_G and T_C cells were detected among the freshly isolated T cells, whereas the T_M cells were detected only following 24 hr of incubation. Removal of T_C cells from the 24-hr-cultured T cells resulted in removal of all the T_C cells and in the concomitant removal of the majority of T_M cells. Similarly, removal of T_M cells from the 24-hr-cultured T cells resulted in the elimination of all T_M cells as well as the majority of T_C cells. These results demonstrate the in vitro generation of T cells with receptors for both FcM and C3 (T_M+C cells). Ten percent of the freshly isolated T_G cells possessed detectable receptors for C3 and/or FcM. These cells constitute the T_G+C and T_G+M lymphocytes. Support for consideration of these receptor-bearing cells as unique and stable cells is provided by the finding that T_M and T_C cells maintained in culture for up to 72 hr do not generate other receptors but retain the single receptor which characterizes each of these cells. Only a small percentage of cultured T_G cells generate receptors for C3 and FcM. It may therefore be concluded that the T_G, T_M, and T_C cells are stable unireceptor-bearing cells. The T_G, T_M, T_C, T_G+C, T_G+M, and T_M+C lymphocytes account for approximately 50% of the circulating lymphocytes. Whether the remaining cells, the T null or T_N cells, constitute the precursors for any or all of the receptor-bearing T cells remains to be determined. Unfractionated freshly isolated T cells were highly cytotoxic in the NK and PWM-mediated MICC assays but were relatively inactive in the ADCC, naturally occurring cell-mediated cytotoxicity (NOCC), and PHA- and Con-A-mediated MICC assays. In contradistinction, T cells incubated for 24 hr displayed marked cytotoxic activity in the ADCC and PHA-mediated MICC assays; they were inactive in the NOCC and Con-Amediated MICC assays. The T_G cells were the predominant cytotoxic cells in the ADCC, NK, and MICC cytotoxic assays since their selective elimination from either the freshly isolated or 24-hr-incubated T cells resulted in almost total loss of cytotoxic activity of the remaining cells. Removal of the T_G+C cells from the freshly isolated or 24-hr-incubated T cells resulted in a significant decrease in PHA- and PWM-mediated MICC cytotoxic activity. T cells depleted of T_M, T_M+C, and T_C cells exhibited the same cytotoxic activity as did the unfractionated T cells. These results suggest that the predominant cytotoxic T cells in all the assays investigated are the T_G cells, that limited cytotoxic activity is also displayed by the T_G+C cells, and that the T_M, T_M+C, T_C, and T_N cells display no cytotoxic activity in the assays utilized in this investigation.     

Structure and Biosynthesis of Cuticular Lipids: Hydroxylation of Palmitic Acid and Decarboxylation of C(28), C(30), and C(32) Acids in Vicia faba Flowers

The structure and composition of the cutin monomers from the flower petals of Vicia faba were determined by hydrogenolysis (LiAlH₄) or deuterolysis (LiAlD₄) followed by thin layer chromatography and combined gas-liquid chromatography and mass spectrometry. The major components were 10, 16-dihydroxyhexadecanoic acid (79.8%), 9, 16-dihydroxyhexadecanoic acid (4.2%), 16-hydroxyhexadecanoic acid (4.2%), 18-hydroxyoctadecanoic acid (1.6%), and hexadecanoic acid (2.4%). These results show that flower petal cutin is very similar to leaf cutin of V. faba. Developing petals readily incorporated exogenous [1-¹⁴C]palmitic acid into cutin. Direct conversion of the exogeneous acid into 16-hydroxyhexadecanoic acid, 10, 16-dihydroxy-, and 9, 16-dihydroxyhexadecanoic acid was demonstrated by radio gas-liquid chromatography of their chemical degradation products. About 1% of the exogenous [1-¹⁴C]palmitic acid was incorporated into C₂₇, C₂₉, and C₃₁n-alkanes, which were identified by combined gas-liquid chromatography and mass spectrometry as the major components of the hydrocarbons of V. faba flowers. The radioactivity distribution among these three alkanes (C₂₇, 15%; C₂₉, 48%; C₃₁, 38%) was similar to the per cent composition of the alkanes (C₂₇, 12%; C₂₉, 43%; C₃₁, 44%). [1-¹⁴C]Stearic acid was also incorporated into C₂₇, C₂₉, and C₃₁n-alkanes in good yield (3%). Trichloroacetate, which has been postulated to be an inhibitor of fatty acid elongation, inhibited the conversion of [1-¹⁴C]stearic acid to alkanes, and the inhibition was greatest for the longer alkanes. Developing flower petals also incorporated exogenous C₂₈, C₃₀, and C₃₂ acids into alkanes in 0.5% to 5% yields. [G-³H]n-octacosanoic acid (C₂₈) was incorporated into C₂₇, C₂₉, and C₃₁n-alkanes. [G-³H]n-triacontanoic acid (C₃₀) was incorporated mainly into C₂₉ and C₃₁ alkanes, whereas [9, 10, 11-³H]n-dotriacontanoic acid (C₃₂) was converted mainly to C₃₁ alkane. Trichloroacetate inhibited the conversion of the exogenous acids into alkanes with carbon chains longer than the exogenous acid, and at the same time increased the amount of the direct decarboxylation product formed. These results clearly demonstrate direct decarboxylation as well as elongation and decarboxylation of exogenous fatty acids, and thus constitute the most direct evidence thus far obtained for an elongation-decarboxylation mechanism for the biosynthesis of alkanes.     

Exohedral complexes of large fullerenes,a theoretical approach

The possibility of existence of exohedral organometallic complexes of fullerenes larger than C₆₀ in which their coordination can have η⁶ hapticity was studied from a theoretical point of view. Complexes containing C₇₀, C₇₄ or C₆₀ cages, as well as cyclopentadienyl (Cp), pentamethyl-cyclopentadienyl (Me₅Cp), benzene rings and hexamethyl-phenyl (Me₆Ph) fragments as ligands, were designed and studied. The results show that many of these molecules can be thermodynamically stable and can have electronic interesting behavior.     

Photosynthesis pathways,ecological characteristics,and the geographical distribution of the Cyperaceae in Japan

Summary The nature of the photosynthetic pathways of Cyperaceae found in Japan were investigated on the basis of Kranz anatomy, the CO₂ compensation concentration and previously reported data. Among 301 species (96% of all cyperaceous species recorded in the region), 58 species were classified as being C₄ plants. These C₄ species were scattered among the tribes Fimbristylideae, Lipocarpheae, Cypereae and Rhynchosporeae in the subfamily Cyperoideae. The genera Cyperus, Eleocharis and Rhynchospora included, in Japan, both C₃ and C₄ species within a single genus. Using these data, an analysis was made of the ecological characteristics and geographical distribution of the C₃ and C₄ species in Japan. Although cyperaceous species grow in markedly different environments, the majority were found in wet and aquatic areas (61%) or shaded areas, such as forest floors (20%). Most of the C₃ species were also hygrophytes (58%) and forest-living species (25%), and C₃ species growing in mesic and dry areas were relatively rare. The C₄ species inhabited wet and aquatic (75%), mesic (13%) and dry areas (6%) and showed marked ecological characteristics with respect to soil-moisture conditions, unlike other C₄ plants, although they were absent from shaded habitats. In order to determine the climatic factors that influence the relative floristic abundance of C₃ and C₄ members of the Cyperaceae in Japan, the ratios of number of C₄ species to the total number of members of Cyperaceae (C₄ percentage) in 16 representative locales were examined in terms of various climatic variables. There were strong positive correlations between the C₄ percentage and temperature. Among the C₃ groups of three subfamilies, there were different distributional trends for various temperature regimes. The C₃ subfamily Caricoideae increased its relative contribution to the cyperaceous flora with a decrease in mean annual temperature, while the C₃ subfamily Sclerioideae exhibited the opposite pattern. The C₃ group of the subfamily Cyperoideae did not show any marked change in pattern along temperature gradients, unlike the two other C₃ subfamilies, and seemed to be heterogeneous in terms of its response to temperature. The relationships between the C₄ biochemical subtypes and ecological characteristics are also discussed.     

Getting the most out of natural variation in C4 photosynthesis

C₄ photosynthesis is a complex trait that has a high degree of natural variation, involving anatomical and biochemical changes relative to the ancestral C₃ state. It has evolved at least 66 times across a variety of lineages and the evolutionary route from C₃ to C₄ is likely conserved but not necessarily genetically identical. As such, a variety of C₄ species are needed to identify what is fundamental to the C₄ evolutionary process in a global context. In order to identify the genetic components of C₄ form and function, a number of species are used as genetic models. These include Zea mays (maize), Sorghum bicolor (sorghum), Setaria viridis (Setaria), Flaveria bidentis, and Cleome gynandra. Each of these species has different benefits and challenges associated with its use as a model organism. Here, we propose that RNA profiling of a large sampling of C₄, C₃–C₄, and C₃ species, from as many lineages as possible, will allow identification of candidate genes necessary and sufficient to confer C₄ anatomy and/or biochemistry. Furthermore, C₄ model species will play a critical role in the functional characterization of these candidate genes and identification of their regulatory elements, by providing a platform for transformation and through the use of gene expression profiles in mesophyll and bundle sheath cells and along the leaf developmental gradient. Efforts should be made to sequence the genomes of F. bidentis and C. gynandra and to develop congeneric C₃ species as genetic models for comparative studies. In combination, such resources would facilitate discovery of common and unique C₄ regulatory mechanisms across genera.     

Urinary selenium in healthy and diabetic Saudi Arabians

We examined Se in urine of 170 Saudi Arabian diabetics (19 insulin-dependent [type 1] and 151 insulin-independent [type 2]) and in an equal number of control subjects of the same origin by measuring the ratio of the concentration of this metal (C_Se) to that of creatinine in urine (C_creat) for each subject. The mean (and SEM) of C_Se/C_creat for the control subjects was 56 (2.9) μmol/mol creat, whereas, the value for the diabetics combined or separated into type 1 and type 2 was 56.7 (3.2), 51.5 (6.3), and 57.4 (3.5) μmol/mol creat, respectively. With the exception of type 2 diabetics who were treated with insulin in addition to oral hypoglycemic and diet (35 patients) (mean [SEM]=43 (4.3) μmol/mol creat), there was no significant difference in C_Se/C_creat between the diabetics and control subjects. Also, there was no significant correlation between C_Se/C_creat and age, sex, or weight of diabetics, whereas, the correlation with the degree of diabetic control was significant (p≤0.0136). Of all diabetes-associated disorders (cardiovascular diseases, neuropathy, ophthalmologic diseases, infections, and hepatic disease), only ophthalmologic diseases appears to cause a significant (p≤0.05) reduction in C_Se/C_creat, but only among type 2 diabetics. Inasmuch as Se status is reflected by urinary Se, healthy Saudi Arabians appear to have Se status that is comparable or higher than those reported for other populations.     

The occurrence of C4 plants and their morphological functional types in the vegetation of Xinjiang, China

Floristic composition, morphological functional types and habitat distributions for C₄ species were studied in Xinjiang, North-western China. 89 species, in 9 families and 41 genera, were identified with C₄ photosynthesis. 48 % of these C₄ species were found in Monocotyledoneae, e.g. Cyperaceae (5 species), Gramineae (38 species), the other 52 % was in Dicotyledoneae, e.g. Chenopodiaceae (29 species), Amaranthaceae (7 species), and Polygonaceae (5 species). Compared with those in semi-arid grasslands in North China and tropical forests in South China, more plant families were found with the occurrence of C₄ plants in this arid region. Relatively higher annual species (63 %), shrubs (18 %), and Chenopodiaceae C₄ species (33 %) compositions were the primary characteristics for the C₄ species occurring in Xinjiang, and this was remarkably related with its arid environment. More Chenopodiaceae C₄ species occurring in the region suggested that this type of C₄ species may have higher capacity to fit the air and soil droughts. There was a strong relationship between C₄ occurrence and habitat distributions, more than half of the total 89 C₄ species were found in disturbed and cultivated lands and early stages of vegetation successions, indicating C₄ occurrence was not only related with climate changes, but also with land uses and vegetation dynamics.     

Neurochemicals for the Investigation of GABA<Subscript>C</Subscript> Receptors

GABA_C receptors are being investigated for their role in many aspects of nervous system function including memory, myopia, pain and sleep. There is evidence for functional GABA_C receptors in many tissues such as retina, hippocampus, spinal cord, superior colliculus, pituitary and the gut. This review describes a variety of neurochemicals that have been shown to be useful in distinguishing GABA_C receptors from other receptors for the major inhibitory neurotransmitter GABA. Some selective agonists (including (+)-CAMP and 5-methyl-IAA), competitive antagonists (such as TPMPA, (±)-cis-3-ACPBPA and aza-THIP), positive (allopregnanolone) and negative modulators (epipregnanolone, loreclezole) are described. Neurochemicals that may assist in distinguishing between homomeric ρ1 and ρ2 GABA_C receptors (2-methyl-TACA and cyclothiazide) are also covered. Given their less widespread distribution, lower abundance and relative structural simplicity compared to GABA_A and GABA_B receptors, GABA_C receptors are attractive drug targets.     

Kinetic analysis of biodesulfurization of model oil containing multiple alkyl dibenzothiophenes

Biodesulfurization is regarded as a promising alternative technology for desulfurization from diesel oil due to its mild operating conditions and its ability to remove sulfur from alky dibenzothiophenes (C_x-DBTs). The diesel oil contains complex mixtures of C_x-DBTs in which individual microbial biodesulfurization may be altered. In this work, interactions among three typical C_x-DBTs such as dibenzothiophenes (DBT), 4-methyldibenzothiophene (4-MDBT), and 4,6-dimethyldibenzothiophene (4,6-DMDBT) were investigated using Mycobacterium sp. ZD-19 in an airlift reactor. The experimental results indicated that the desulfurization rates would decrease in the multiple C_x-DBTs system compared to the single C_x-DBT system. The extent of inhibition depended upon the substrate numbers, concentrations, and affinities of the co-existing substrates. For example, compared to individual desulfurization rate (100 %), DBT desulfurization rate decreased to 75.2 % (DBT + 4,6-DMDBT), 64.8 % (DBT + 4-MDBT), and 54.7 % (DBT + 4,6-DMDBT + 4-MDBT), respectively. This phenomenon was caused by an apparent competitive inhibition of substrates, which was well predicted by a Michaelis–Menten competitive inhibition model.     

Cell Envelope Phospholipid Composition of Burkholderia multivorans

Burkholderia multivorans causes opportunistic pulmonary infections in cystic fibrosis and immunocompromised patients. The purpose of the present study was to determine the nature of the phospholipids and their fatty acid constituents comprising the cell envelope membranes of strains isolated from three disparate sources. A conventional method for obtaining the readily extractable lipids fraction from bacteria was employed to obtain membrane lipids for thin-layer chromatographic and gas chromatography-mass spectrophotometric analyses. Major fatty acid components of the B. multivorans readily extractable lipid fractions included C_16:0 (palmitic acid), C_16:1 (palmitoleic acid), and C_18:1 (oleic acid), while C_14:0 (myristic acid), ΔC_17:0 (methylene hexadecanoic acid), C_18:0 (stearic acid), and ΔC_19:0 (methylene octadecanoic acid) were present in lesser amounts. Fatty acid composition differed quantitatively among strains with regard to C_16:0, C_16:1, ΔC_17:0, C_18:1, and ΔC_19:0 with the unsaturated:saturated fatty acid ratios being significantly less in a cystic fibrosis type strain than either environmental or chronic granulomatous disease strains. Phospholipids identified in all B. multivorans strains included lyso-phosphatidylethanolamine, phosphatidylglycerol, phosphatidylethanolamine, and diphosphatidylglycerol in similar ratios. These data support the conclusion that the cell envelope phospholipid profiles of disparate B. multivorans strains are similar, while their respective fatty acyl substituent profiles differ quantitatively under identical cultivation conditions.     

Occurrence and Some Properties of Cellulase in the Filtrates of Conidiospores and Mycelia of Pyricularia oryzae Cavara

Occurrence of cellulase activity was demonstrated in the filtrates of germinating conidiospores and growing mycelia of P. oryzae. Activity and some properties of cellulase in the filtrate of mycelia grown on rice plant powder as carbon source were compared among various strains.

Cellulase activity (C₁ and C_x enzymes; cellulose and carboxymethylcellulose as substrates, respectively) in the filtrate of germinating conidiospores was detected in the pathogenic T–l (Ken 53–33) strain as well as nonpathogenic 0 (THU 3 × 1) strain of P. oryzae. The activity was higher in the former than the latter strains. Cellulase activity (C_x enzyme) in the filtrate of growing mycelia was detected in the four strains used, T–l (Ken 53–33), C–3 (N 87), N–1 (H373), and 0 (THU 3 × 1). Cellulase activity (C_x enzyme) in the filtrate of mycelia was optimal at pH 5.0 and 40°C, and stable up to 40°C. Their properties did not differ significantly except for the pH-activity curve at alkaline side among various strains; but cellulase activity (C₁ enzyme) was found to be correlated with their pathogenicity except for the case of C–3 strain.     

The occurrence of C4 photosynthesis in Yunnan province, a tropical region in South-western China

Floristic composition, morphological functional types, and altitudinal distribution pattern for C₄ species were studied in Yunnan province, South-western China. 159 species, in 6 families and 60 genera, were identified with C₄ photosynthesis. 93 % of these C₄ species were found in Monocotyledoneae, e.g. Cyperaceae (18 species), Gramineae (129 species), and Commelinaceae (1 species), the other 7 % was in Dicotyledoneae, e.g. Amaranthaceae (5 species), Portulacaceae (4 species), and Chenopodiaceae (2 species). Hence C₄ plants mainly occurred in very few families in the tropical region. Compared with those in semi-arid grasslands and arid deserts in North China, more C₄ grasses and much less Chenopodiaceae C₄ species occurred in the tropical region. This indicates the physiological responses of C₄ plants from the two families are very different. Chenopodiaceae C₄ species may be more fit semi-arid and arid environments, while C₄ grasses are more fit the moist tropical conditions. There was a strong relationship between C₄ distribution and altitude in the tropical region. Altitudinal distribution pattern for C₄ species in the region was consistent with altitude, climate, and habitats.     

Application of Plasma Levels of Olanzapine and N-Desmethyl-Olanzapine to Monitor Clinical Efficacy in Patients with Schizophrenia

Background

This therapeutic drug monitoring (TDM) study aimed to determine the role of olanzapine (OLZ) and N-desmethyl-OLZ (DMO) levels in the therapeutic efficacy of OLZ in patients with schizophrenia.

Method

Plasma concentrations of OLZ (C_OLZ) and DMO (C_DMO) in schizophrenic patients 12 hours post-dose were assessed. The correlations of C_OLZ and C_DMO with the various scores of the Positive and Negative Syndrome Scale (PANSS) were evaluated. A receiver operating characteristic curve (ROC) was utilized to identify the threshold C_OLZ and C_OLZ/C_DMO ratio for maintenance of satisfactory efficacy.

Results

A total of 151 samples from patients with schizophrenia were analyzed for individual C_OLZ and C_DMO levels. The mean C_OLZ and C_DMO levels were 37.0 ± 25.6 and 6.9 ± 4.7 ng/mL, respectively, and C_OLZ was ~50% higher in female or nonsmokers (p<0.01). In all patients, the daily dose of OLZ was positively correlated with C_OLZ and C_DMO. Linear relationships between C_OLZ and OLZ dose were observed in both nonsmokers and smokers (r_s = 0.306, 0.426, p<0.01), although C_DMO was only correlated with OLZ dose in smokers (r_s = 0.485, p<0.01) and not nonsmokers. In all patients, C_OLZ was marginally negatively correlated with the total PANSS score. The total PANSS score was significantly negatively correlated with the C_OLZ/C_DMO ratio (p<0.005), except in smokers. The ROC analysis identified a C_OLZ/C_DMO ratio ≥2.99 or C_OLZ ≥22.77 ng/mL as a predictor of maintenance of an at least mildly ill status (PANSS score ≤58) of schizophrenia in all patients.

Conclusions

A significantly negative correlation between the steady-state C_OLZ/C_DMO ratio and total PANSS score was observed in Taiwanese schizophrenic patients. TDM of both OLZ and DMO levels could assist clinical practice when individualizing OLZ dosage adjustments for patients with schizophrenia.     

Global remote sensing of water–chlorophyll ratio in terrestrial plant leaves

I evaluated the use of global remote sensing techniques for estimating plant leaf chlorophyll a + b (C_ab; μg cm⁻²) and water (C_w; mg cm⁻²) concentrations as well as the ratio of C_w/C_ab with the PROSAIL model under possible distributions for leaf and soil spectra, leaf area index (LAI), canopy geometric structure, and leaf size. First, I estimated LAI from the normalized difference vegetation index. I found that, at LAI values <2, C_ab, C_w, and C_w/C_ab could not be reliably estimated. At LAI values >2, C_ab and C_w could be estimated for only restricted ranges of the canopy structure; however, the ratio of C_w/C_ab could be reliably estimated for a variety of possible canopy structures with coefficients of determination (R²) ranging from 0.56 to 0.90. The remote estimation of the C_w/C_ab ratio from satellites offers information on plant condition at a global scale.     

A comparative immunocytochemical localization study of phosphoenolpyruvate carboxylase in leaves of higher plants

The intracellular localization of phosphoenolpyruvate (PEP) carboxylase in plants belonging to the C₄, Crassulacean acid metabolism (CAM) and C₃ types was invetigated using an immunocytochemical method with an immune serum raised against the sorghum leaf enzyme. The plants studied were sorghum, maize (C₄ type), kalanchoe (CAM type), french bean, and spinach (C₃ type). In the green leaves of C₄ plants, it was shown that the carboxylase was located in the mesophyll and stomatic cells, being largely cytosolic in the mesophyll cells. Similarly, in CAM plants, the enzyme was found mainly outside the chloroplasts. In contrast, in C₃ plants, the PEP carboxylase appeared to be distributed between the cytosol and the chloroplasts of foliar parenchyma. Examination of sections from etiolated leaves showed fluorescence emission from etioplasts and cytosol for the parenchyma of french bean as well as for the bundle sheath and mesophyll of sorghum leaves. This data indicated that during the greening process photoregulation and evolution of PEP carboxylase is dependent on the tissue and on the metabolic type of the plant considered.Abbreviations CAM Crassulacean acid metabolism - PEP phosphoenolpyruvate     

Neuroprotective Effects of Butanol Fraction of Cordyceps cicadae on Glutamate‐Induced Damage in PC12 Cells Involving Oxidative Toxicity

The current study was aimed at investigating the neuroprotective effects of the butanol fraction from Cordyceps cicadae (C_BU), which was responsible for the anti‐aging effect of this medicine. Glutamate‐induced PC12 cells were used as a model to determine the neuroprotective effect against oxidative cell death. Cell viability, cytotoxicity, flow cytometry, mitochondrial transmembrane potential (MMP), reactive oxygen species (ROS), glutathione peroxidase (GSH‐Px), and superoxide dismutase (SOD) levels were analyzed to assess neuronal cell survival or death. The results obtained from the above evaluations showed that C_BU was the most effective fraction and even better than pure compounds present in C. cicadae in terms of suppressing glutamate‐induced damage in PC12 cells, increasing cell viability, decreasing lactase dehydrogenase (LDH) release, and reduction of apoptosis induced by exposure to glutamate. Furthermore, C_BU protected cells against mitochondrial dysfunction and oxidative stress as indicated by the suppression of ROS accumulation and up regulation of the levels of GSH‐Px and SOD. In summary, the above results showed that C_BU exerted neuroprotective effect against oxidative damage, and this activity could be partly due to the action of nucleosides present in the C_BU.     

Lisianthus response to salinity stress

The effect of salinity on some morpho-physiological characteristics in lisianthus cultivars was investigated. Cultivars namely, Blue Picotee (C1), Champagne (C₂), Lime Green (C₃), and Pure White (C₄), were subjected to salt stress (0–60 mM NaCl) in a sand culture and their responses were measured. Our results showed that as a salinity level increased, growth parameters, relative water content, photosynthetic pigments, and gas-exchange characteristics decreased in all cultivars, while root fresh mass, root/shoot length ratio, electrolyte leakage, and a malondialdehyde content increased. However, the changes were less pronounced in C₃ and C₄ compared to C1 and C₂. The regression analysis of the relationship between salinity levels and seedling height or root/shoot length ratio defined two groups with different slope coefficients: C1 and C₂ as salt-sensitive cultivars and C₃ and C₄ as salt-tolerant cultivars. Shoot dry mass and leaf area tolerance indices were less affected by salinity in C₃ and C₄ compared to those in C1 and C₂. Further, C₃ and C₄ showed higher photosynthetic rates, greater stomatal conductances, and accumulated greater K+ and Ca2+ contents and K+/Na+ ratios in roots and shoots compared to those in C1 and C₂. The results suggests that C₃ and C₄ could be recommended as resistant cultivars due to maintaining higher growth, water balance, leaf gas exchange, ion compartmentalization, and lower lipid peroxidation in response to salinity compared to C1 and C₂.     

Effect of inland salt-alkaline stress on C4 enzymes, pigments, antioxidant enzymes, and photosynthesis in leaf, bark, and branch chlorenchyma of poplars

The effects of soil salt-alkaline (SA) stress on leaf physiological processes are well studied in the laboratory, but less is known about their effect on leaf, bark and branch chlorenchyma and no reports exist on their effect on C₄ enzymes in field conditions. Our results demonstrated that activities of C₄ enzymes, such as phospholenolpyruvate carboxylase (PEPC), NADP-malic enzyme (NADP-ME), pyruvate orthophosphate dikinase (PPDK), and NADP-dependent malate dehydrogenase (NADP-MDH), could also be regulated by soil salinity/alkalinity in poplar (Populus alba × P. berolinensis) trees, similarly as the already documented changes in activities of antioxidative enzymes, such as superoxide dismutase (SOD), catalase (CAT), and glutathione reductase (GR), pigment composition, photosynthesis, and respiration. However, compared with 50–90% changes in a leaf and young branch chlorenchyma, much smaller changes in malondialdehyde (MDA), antioxidative enzymes, and C₄ enzymatic activities were observed in bark chlorenchyma, showing that the effect of soil salinity/alkalinity on enzymatic activities was organ-dependent. This suggests that C₄ enzymatic ratios between nonleaf chlorenchyma and leaf (the commonly used parameter to discern the operation of the C₄ photosynthetic pathway in nonleaf chlorenchyma), were dependent on SA stress. Moreover, much smaller enhancement of these ratios was seen in an improved soil contrary to SA soil, when the fresh mass (FM) was used as the unit compared with a calculation on a chlorophyll (Chl) unit. An identification of the C₄ photosynthesis pathway via C₄ enzyme difference between chlorenchyma and leaf should take this environmental regulation and unit-based difference into account.     

Epicuticular wax of Agropyron intermedium

Wax on leaves of Agropyron intermedium contains hydrocarbons (11%, C₂₇–C₃₃), esters (11%, C₃₂–C₆₀), free alcohols (180%, C₂₆) 25-oxohentriacontane-14,16-dione (17%), 10-oxohentriacontane-14,16-dione (5y%), 25-hydroxyhentriacontane-14,16-dione (12%) and 26-hydroxyhentriacontane-14,16-dione (2%). Wax on spikes contains additional components, C₂₅–C₃₃cis 9-alkenes (32% of hydrocarbons), and more β-diketones, 25-hydroxy (17%) and 26-hydroxy (3%) hentriacontane-14,16-diones, 10,25-dioxohentriacontane-14,16-dione (1%) and 4-hydroxy-25-oxo-(2%), 25-hydroxy-10-oxo-(1.3%) and 26-hydroxy-10-oxo-(0.7%) hentriacontane-14,16-diones; free alcohols were very minor components (1%, C₂₄–C₃₂).