Accumulation Of Calcium Oxalate Crystals Research Articles

The Study of Crystals in the Fruits of Some Apiaceae Species Using Energy-Dispersive Spectroscopy

Crystals in the fruits of Apiaceae are of great importance for taxonomy. The presence/absence and location of large prismatic crystals, druses, and spherical aggregates characterize genera, tribes, and subfamilies. When the fruits of Apiaceae split into mericarps, cell separation occurs, and probably cell death and the destruction of their walls. In clades Saniculeae and Scandicinae, this process is accompanied by the accumulation of calcium oxalate crystals in the abscission layer. However, reports of smaller crystals are rare, and their taxonomic significance is unknown. To fill this gap, we started research with four species of the clade Apieae: Ammi majus, Modesciadium involucratum, Deverra aphylla, and Rutheopsis tortuosa; in the last two species, crystals were described for the first time. We also studied crystals in Chaerophyllum bulbosum from the clade Scandicinae, and in Sanicula rubriflora from the clade Saniculeae. Conventional optical and polarizing microscopy, scanning electron microscopy, and energy dispersive spectroscopy (EDS) were used. In the clade Apieae, the studied species contained crystals in the exocarp and on the commissure, and often in the mesocarp parenchyma as well; the shapes of the crystals are small druses and crystal sand. The calcium content in the crystals was determined as 10–31%; crystals contain also carbon and oxygen and are most likely composed of calcium oxalate. In the fruits of Apiaceae studied, the silicon content is low (<0.3%).

A sulfated polysaccharide κ-carrageenan induced antioxidant defense and proteomic changes in chloroplast against leaf spot disease of tomato

Ecofriendly approaches to control plant diseases using biopolymers are gaining momentum due to their non-toxic nature to the environment. The present study elucidates the potency of a sulfated polysaccharide, κ-carrageenan, obtained from the red seaweed Kappaphycus alvarezii, as an inducer of antioxidant defense and also chloroplast proteome against leaf spot disease in tomato. к-carrageenan (0.3%) treatment was very effective in reducing disease scores in tomato plants. Fluorescent microscopy of tomato leaves stained with Uvitex 2B (a chitin specific stain) revealed that foliar application of κ-carrageenan (0.3%) reduced colonization of the pathogen Septoria lycopersici. In addition, κ-carrageenan pretreatment followed by pathogen infection recorded a higher accumulation of calcium oxalate crystals. The treatment of κ-carrageenan resulted in the induction of H2O2 and superoxide anion compared to control or S. lycopersici-infected plants. The tomato leaves treated with κ-carrageenan showed higher activity of peroxidase enzyme. Furthermore, chloroplast proteome analysis by 2D-PAGE of control, S. lycopersici-infected, κ-carrageenan-treated, and к-carrageenan treatment followed by S. lycopersici-infected leaves of tomato showed differential expression of 31 proteins, of which 11 proteins were upregulated including acid phosphatase 1-like, C- terminal processing peptidase, GATA transcription factor 5, Calvin cycle protein CP12-1, Photosystem I reaction center subunit PSAK, and subunit of cytochrome B6-F complex iron-sulfur centers. These results indicated that κ-carrageenan could be a promising inducer of antioxidant defense and modulator of chloroplast proteome in tomato plants.

Small Molecule-Based Enzyme Inhibitors in the Treatment of Primary Hyperoxalurias.

Primary hyperoxalurias (PHs) are a group of inherited alterations of the hepatic glyoxylate metabolism. PHs classification based on gene mutations parallel a variety of enzymatic defects, and all involve the harmful accumulation of calcium oxalate crystals that produce systemic damage. These geographically widespread rare diseases have a deep impact in the life quality of the patients. Until recently, treatments were limited to palliative measures and kidney/liver transplants in the most severe forms. Efforts made to develop pharmacological treatments succeeded with the biotechnological agent lumasiran, a siRNA product against glycolate oxidase, which has become the first effective therapy to treat PH1. However, small molecule drugs have classically been preferred since they benefit from experience and have better pharmacological properties. The development of small molecule inhibitors designed against key enzymes of glyoxylate metabolism is on the focus of research. Enzyme inhibitors are successful and widely used in several diseases and their pharmacokinetic advantages are well known. In PHs, effective enzymatic targets have been determined and characterized for drug design and interesting inhibitory activities have been achieved both in vitro and in vivo. This review describes the most recent advances towards the development of small molecule enzyme inhibitors in the treatment of PHs, introducing the multi-target approach as a more effective and safe therapeutic option.

Sulfamethoxazole activates antioxidant defense response against Alternaria solani infection in Solanum lycopersicum Linn

The present study focuses on the potency of sulfamethoxazole (Sm) on inducing resistance against Alternaria solani infection. Anatomical observation indicated calcium oxalate deposits in pathogen-infected tissue compared to control plants. Sm pretreated and pathogen-infected tomato leaves showed decreased accumulation of calcium oxalate crystals. The oxalic acid content is found decreased in Sm (3 µM) pretreated and pathogen-infected leaves at 36 h compared to control and pathogen-infection (PI). Sm treatment resulted in transient and significant increase in H2O2 level during early stages of PI. A significant increase in superoxide dismutase, guaiacol dependent peroxidase and polyphenol oxidase was evident in Sm treated leaves which altogether contributed to protection of tomato from oxidative stress due to PI. Sm treatment followed by PI resulted in accumulation of superoxide anion compared to A. solani infected plants. In conclusion, Sm treatment regulate balance between antioxidants and free radicals, induces antioxidant enzymes and thereby controlling pathogen ingress.

Effect of Acyl Activating Enzyme (AAE) 3 on the growth and development of Medicago truncatula

The Acyl-Activating Enzyme (AAE) 3 gene encodes an oxalyl-CoA synthetase that catalyzes the conversion of oxalate to oxalyl-CoA in a CoA and ATP-dependent manner. Although the biochemical activity of AAE3 has been established, its biological role in plant growth and development remains unclear. To advance our understanding of the role of AAE3 in plant growth and development, we report here the characterization of two Medicago truncatula AAE3 (Mtaae3) mutants. Characterization of a Mtaae3 RNAi mutant revealed an accumulation of calcium oxalate crystals and increased seed permeability. These phenotypes were also exhibited in the Arabidopsis aae3 (Ataae3) mutants. Unlike the Ataae3 mutants, the Mtaae3 RNAi mutant did not show a reduction in vegetative growth, decreased seed germination, or increased seed calcium concentration. In an effort to clarify these phenotypic differences, a Mtaae3 Tnt1 mutant was identified and characterized. This Mtaae3 Tnt1 mutant displayed reduced vegetative growth, decreased seed germination, and increased seed calcium concentration as well as an accumulation of calcium oxalate crystals and increased seed permeability as found in Ataae3. Overall, the results presented here show the importance of AAE3 in the growth and development of plants. In addition, this study highlights the ability to separate specific growth and development phenotypes based on the level of AAE3 gene expression.

Salicylic Acid Derivatives Inhibit Oxalate Production in Mouse Hepatocytes with Primary Hyperoxaluria Type 1

Primary hyperoxaluria type 1 (PH1) is a rare life-threatening genetic disease related to glyoxylate metabolism and characterized by accumulation of calcium oxalate crystals. Current therapies involve hepatic and/or renal transplantation, procedures that have significant morbidity and mortality and require long-term immunosuppression. Thus, a pharmacological treatment is urgently needed. We introduce here an unprecedented activity of salicylic acid derivatives as agents capable of decreasing oxalate output in hyperoxaluric hepatocytes at the low micromolar range, which means a potential use in the treatment of PH1. Though correlation of this phenotypic activity with glycolate oxidase (GO) inhibition is still to be verified, most of the salicylic acids described here are GO inhibitors with IC50 values down to 3 μM. Binding mode of salicylic acids inside GO has been studied using in silico methods, and preliminary structure-activity relationships have been established. The drug-like structure and ease of synthesis of our compounds make them promising hits for structural optimization.

Does aridity influence the morphology, distribution and accumulation of calcium oxalate crystals in Acacia (Leguminosae: Mimosoideae)?

Calcium oxalate (CaOx) crystals are a common natural feature of many plant families, including the Leguminosae. The functional role of crystals and the mechanisms that underlie their deposition remain largely unresolved. In several species, the seasonal deposition of crystals has been observed. To gain insight into the effects of rainfall on crystal formation, the morphology, distribution and accumulation of calcium oxalate crystals in phyllodes of the leguminous Acacia sect. Juliflorae (Benth.) C. Moore & Betche from four climate zones along an aridity gradient, was investigated. The shapes of crystals, which include rare Rosanoffian morphologies, were constant between species from different climate zones, implying that morphology was not affected by rainfall. The distribution and accumulation of CaOx crystals, however, did appear to be climate-related. Distribution was primarily governed by vein density, an architectural trait which has evolved in higher plants in response to increasing aridity. Furthermore, crystals were more abundant in acacias from low rainfall areas, and in phyllodes containing high concentrations of calcium, suggesting that both aridity and soil calcium levels play important roles in the precipitation of CaOx. As crystal formation appears to be calcium-induced, we propose that CaOx crystals in Acacia most likely function in bulk calcium regulation.

Acúmulo de ácido oxálico e cristais de cálcio em ectomicorrizas de eucalipto.: II- formação de cristais de oxalato de cálcio induzida por fungos ectomicorrízicos em raízes laterais finas

O eucalipto é eficiente na aquisição de Ca do solo, mas pouco se sabe sobre a participação das ectomicorrizas e dos ácidos orgânicos nesse processo em campo. O acúmulo de cristais de Ca (CaOx) foi avaliado em, aproximadamente, 2.100 raízes laterais finas e ectomicorrizas do híbrido de Eucalyptus grandis x Eucalyptus urophylla, cultivado por 2,5 anos em área com topografia típica em meia laranja, com vertente côncavo-convexa, na região de Viçosa, MG. Técnicas de microscopia óptica e microscopia eletrônica de varredura foram usadas para a visualização dos CaOx. Em 73,7 % das raízes, ocorreu abundante acúmulo de drusas e grânulos de CaOx nas células do córtex. A presença conspícua de CaOx foi observada em 56,2 % das ectomicorrizas e em 17,5 % das raízes laterais finas não colonizadas, evidenciando o papel das micorrizas no acúmulo de Ca em eucalipto. A forma predominante dos CaOx foram as drusas nas ectomicorrizas e os grânulos cristalinos nas raízes. Os dez morfotipos de ectomicorrizas observados na área diferiram quanto à presença e à morfologia dos CaOx, o que pode representar distintas capacidades dos fungos ectomicorrízicos em fornecer Ca para a planta hospedeira. A análise da superfície do manto das ectomicorrizas por microscopia eletrônica de varredura não evidenciou a presença de CaOx nessa estrutura, confirmando que, nas condições avaliadas, o acúmulo de cristais limita-se ao córtex radicular. Este é o primeiro relato da ocorrência de CaOx em ectomicorrizas de eucalipto no Brasil, com dados que comprovam que há mecanismos de armazenamento de Ca nas ectomicorrizas em áreas com baixa disponibilidade do elemento.

Effects of the Papaya meleira virus on papaya latex structure and composition

Spontaneous latex exudation is the main symptom of papaya sticky (meleira) disease caused by the Papaya meleira virus (PMeV), a double-stranded RNA (dsRNA) virus. This paper describes different effects of PMeV on papaya latex. Latex samples were subjected to different histochemical tests to evaluate their chemical composition. Additionally, the integrity of the latex particles was assessed by transmission and scanning electron microscopy analysis. Biochemical and micro- and macro-element measurements were performed. PMeV dsRNA extraction was performed to evaluate the interaction of the virus with the latex particles. Sticky diseased latex was positive for alkaloid biosynthesis and showed an accumulation of calcium oxalate crystals. PMeV also increased H(2)O(2) synthesis within sticky diseased laticifers. The protein, sugar and water levels were altered, probably due to chemical changes. The morphology of the latex particles was further altered; PMeV particles seemed to be bound to the latex particles. The alkaloid and H(2)O(2) biosynthesis in the papaya laticifers indicate a papaya defense response against PMeV. However, such efforts failed, as the virus affected the plant latex. The effects described here suggest some advantages of the infection process, including facilitating the movement of the virus within the papaya plant.

Physiological and chemical characteristics of field-and mountain-cultivated ginseng roots

Demand is increasing for mountain-cultivatedPanax ginseng (MCG) because its quality is considered superior to that of field-cultivated ginseng (FCG). However, MCG grows very slowly, and the factors that might affect this are unknown. In addition, little information is available about the physiological characteristics of its roots. Here, we investigated local soil environments and compared the histological and chemical properties of MCG and FCG roots. Average diameters, lengths, and fresh weights were much smaller in the former. Photosynthesis rates and root cambial activity also were reduced in the MCG tissues. Our analysis of soil from the mountain site revealed an extremely low phosphorus content, although those samples were richer in total nitrogen and organic matter than were the field soils. MCG roots also contained higher amounts of ginsenosides, and total accumulations increased with age. Moreover, ginsenoside Rh2, a red ginseng-specific compound, accumulated in the MCG roots but not in those from FCG plants. Interestingly, numerous calcium oxalate crystals were found in MCG roots, particularly in their rhizomes (i.e., short stems). Therefore, we can conclude from these results that low levels of the essential mineral phosphorus in mountain soils are a critical factor that retards the growth of mountain ginseng. Likewise, the high accumulation of calcium oxalate crystals in MCG roots might be an adaptation mechanism for survival in such a harsh local environment.

Morpho-anatomical changes and photosynthetic metabolism of Stenocereus beneckei seedlings under soil water deficit

Characteristics developed by Cactaceae for adaptation to climates where water is limited include crassulacean acid metabolism (CAM), a thick cuticle, and spines and trichomes that intercept a proportion of solar radiation. A few studies consider morpho-anatomical and physiological characteristics of Cactaceae seedlings, which may help understand their establishment, growth, and eventual reproduction. In this study, photosynthetic metabolism (titratable protons) and morpho-anatomical features of Stenocereus beneckei seedlings were examined under limiting water conditions. Soil moisture treatments consisted of -0.03, -0.5, -1.5, and -3.0 MPa, and seedling samples were taken at 3 h intervals on one day at 7 and 9 months of age with three replicates per treatment. The results show irregular fluctuations in acidity concentrations during the first 6 and 7 months of age; at 9 months, an increase in titratable proton values was observed during the night, and it seems that soil moisture does not determine CAM expression. Seedlings from smaller seeds are less tolerant to water stress, they had poor growth in all treatments, and at -3.0 MPa after 3 months of drought none survived. Anatomical observations show collapsed cells associated with a high accumulation of calcium oxalate crystals and starch grains, as a response to water deficit. Titratable acidity concentration increased with seedling age, and CAM expression did not accelerate with soil water deficit.

Morphological Changes in Leaves of Mexican Lime Affected by Iron Chlorosis

ABSTRACT Iron (Fe) chlorosis reduces the concentration of photosynthetic pigments, photosynthates, and crop yield. The effect of Fe chlorosis on leaf composition and cell structure was evaluated in Mexican lime (Citrus aurantifolia) with different degrees of Fe chlorosis. Iron chlorosis significantly reduced concentrations of chlorophylls a, b, and a + b, and caused thickening of leaves, due to the increase in palisade and spongy parenchyma cells. The chloroplasts of the chlorotic and albino leaves showed a disorganized ultrastructure; they had an elongated shape with disarrayed thylakoids, underdeveloped grana, scarce starch granules, and hole-like folds in the thylakoid membranes. The accumulation of calcium oxalate crystals in the upper and lower sides of the epidermis, crystal length, and total crystal content increased with Fe chlorosis severity. The green leaves, in contrast, had chloroplasts with typical ultrastructure. The degree of Fe chlorosis in the leaves significantly affected the concentrations of potassium (K); Fe, manganese (Mn), Fe2+, and the phosphorus (P)/Fe and K/calcium (Ca) ratios.

Primary hyperoxaluria in an adult with renal failure, livedo reticularis, retinopathy, and peripheral neuropathy

We present the case of a young woman who developed renal failure of unknown cause, and after 2 months of maintenance hemodialysis developed livedo reticularis, retinopathy, and peripheral sensory neuropathy. The patient was subsequently shown to have primary oxalosis type I, a rare autosomal recessive error of metabolism characterized by accumulation of calcium oxalate crystals in the kidneys, eyes, skin, and other organs. Intravascular obstruction, caused by deposition of calcium oxalate crystals in cutaneous arterioles, is thought to be responsible for the ischemic livedo reticularis lesions observed in this patient. A method is described for measuring serum glycolate by isotope dilution gas chromatography-mass spectrometry (GC-MS). An approach to the diagnosis and management is also briefly mentioned.

Primary oxalosis-an unusual cause of livedo reticularis

A young woman presented with renal failure and skin lesions of livedo reticularis (LR) due to primary oxalosis. Primary oxalosis is a rare autosomal-recessive error of metabolism characterized by accumulation of calcium-oxalate crystals in the kidneys, eyes, heart and skin. This unusual cause of intravascular obstruction resulting in livedo reticularis should be considered in patients with renal impairment. An approach to the diagnosis of LR is presented.

Cardiac sarcoplasmic reticulum: Chemical and electron microscope studies of calcium accumulation

A sucrose gradient preparation (35–45 % sucrose layer) of fragmented sarcoplasmic reticulum (FSR) from canine myocardium has been examined by electron microscopy after ATP-induced accumulation of calcium oxalate crystals within vesicles. In sectioned pellets, the loaded vesicles appear centrifugally concentrated in the lowermost 10% of the pellet thickness. The high ratio of empty to full vesicles was also observed in suspensions deposited on electron microscope grids, examined without staining. Furthermore, black calcium oxalate crystals were produced in vesicles, adsorbed to the carbon foil, by floating the grid on a drop of incubation medium. Centrifugal isolation of loaded vesicles has produced a FSR fraction with more than half of the vesicles containing calcium oxalate. The enriched pellets contain only 15–20% of the original FSR protein in association with 80–90% of the total calcium from the incubation mixture, or 22–26 μmoles Ca/mg FSR protein. ATPase activity demonstrable with the loaded vesicle fraction corresponded to 45% of the original total FSR activity, including calcium-activated “extra ATPase” corresponding to 22–33 % of that in the original total FSR, while the specific activity was increased 2- and 3-fold, respectively.