Complement Receptor C3aR1 Contributes to Paclitaxel-Induced Peripheral Neuropathic Pain in Mice and Rats.

Cancer chemotherapy-induced neuropathic pain is a devastating pain syndrome without effective therapies. We previously reported that rats deficient in complement C3, the central component of complement activation cascade, showed a reduced degree of paclitaxel-induced mechanical allodynia (PIMA), suggesting that complement is integrally involved in the pathogenesis of this model. However, the underlying mechanism was unclear. Complement activation leads to the production of C3a, which mediates inflammation through its receptor C3aR1. In this article, we report that the administration of paclitaxel induced a significantly higher expression level of C3aR1 on dorsal root ganglion (DRG) macrophages and expansion of these macrophages in DRGs in wild-type (WT) compared with in C3aR1 knockout (KO) mice. We also found that paclitaxel induced less severe PIMA, along with a reduced DRG expression of transient receptor potential channels of the vanilloid subtype 4 (TRPV4), an essential mediator for PIMA, in C3aR1 KO than in WT mice. Treating WT mice or rats with a C3aR1 antagonist markedly attenuated PIMA in association with downregulated DRG TRPV4 expression, reduced DRG macrophages expansion, suppressed DRG neuron hyperexcitability, and alleviated peripheral intraepidermal nerve fiber loss. Administration of C3aR1 antagonist to TRPV4 KO mice further protected them from PIMA. These results suggest that complement regulates PIMA development through C3aR1 to upregulate TRPV4 on DRG neurons and promote DRG macrophage expansion. Targeting C3aR1 could be a novel therapeutic approach to alleviate this debilitating pain syndrome.

A mutant cotton fatty acid desaturase 2-1d allele causes protein mistargeting and altered seed oil composition.

BACKGROUND: Cotton (Gossypium sp.) has been cultivated for centuries for its spinnable fibers, but its seed oil also possesses untapped economic potential if, improvements could be made to its oleic acid content. RESULTS: Previous studies, including those from our laboratory, identified pima accessions containing approximately doubled levels of seed oil oleic acid, compared to standard upland cottonseed oil. Here, the molecular properties of a fatty acid desaturase encoded by a mutant allele identified by genome sequencing in an earlier analysis were analyzed. The mutant sequence is predicted to encode a C-terminally truncated protein lacking nine residues, including a predicted endoplasmic reticulum membrane retrieval motif. We determined that the mutation was caused by a relatively recent movement of a Ty1/copia type retrotransposon that is not found associated with this desaturase gene in other sequenced cotton genomes. The mutant desaturase, along with its repaired isozyme and the wild-type A-subgenome homoeologous protein were expressed in transgenic yeast and stably transformed Arabidopsis plants. All full-length enzymes efficiently converted oleic acid to linoleic acid. The mutant desaturase protein produced only trace amounts of linoleic acid, and only when strongly overexpressed in yeast cells, indicating that the missing C-terminal amino acid residues are not strictly required for enzyme activity, yet are necessary for proper subcellular targeting to the endoplasmic reticulum membrane. CONCLUSION: These results provide the biochemical underpinning that links a genetic lesion present in a limited group of South American pima cotton accessions and their rare seed oil oleic acid traits. Markers developed to the mutant desaturase allele are currently being used in breeding programs designed to introduce this trait into agronomic upland cotton varieties.

Defending Effects of Iodide Transfer in Placental Barrier Against Maternal Iodine Deficiency.

Objective: Placental iodide transport is necessary for maintaining an adequate iodide supply to the developing fetus. We hypothesized that compounds from the placental barrier can compensate for decreases in maternal iodine intake and normalize fetal iodine levels. Methods: Pregnant rats administered different amounts of iodine (1.24, 2.5, 5, or 10 µg/day) were evaluated on gestational day (gd) 16 and 20. The iodine levels in maternal blood, amniotic fluid (AF), and placental tissue were estimated using As-Ce catalytic spectrophotometry. The protein and/or messenger RNA (mRNA) levels of sodium iodide symporter (NIS), pendrin, alpha-smooth muscle actin (α-SMA), and CD31 in the placental labyrinth, trophoblast cells isolated using laser capture microdissection (LCM), and/or fetomaternal thyroid were detected using immunoblotting, real-time polymerase chain reaction, and/or immunohistochemistry. Results: When iodine intake was reduced, iodine levels in maternal blood gradually decreased; however, placental iodine levels were not significantly different between groups on gd16 and gd20. Minimal changes were observed in AF iodine levels on gd16, and a mild decreasing trend was observed (iodine dose, 10 to 1.24 µg/day) on gd20. NIS protein, which was linearly distributed along the basolateral membrane of maternal-fetal thyroid follicles, gradually increased with decreasing iodine levels. Regarding iodine deficiency in the placental labyrinth on gd16 and gd20, pendrin and glycosylated NIS proteins were significantly upregulated in a dose-dependent manner. However, the mRNA levels were unchanged. Furthermore, the conversion of NIS protein from the nonglycosylated to the glycosylated form increased. In trophoblast cells isolated using LCM, PDS mRNA levels increased in the 1.24-µg/day group on gd16 but not NIS mRNA levels. There was a smaller α-SMA+ area in the labyrinth zone on gd16 and gd20; however, the proportional CD31+ area increased on gd16 and reduced on gd20 with decreased iodine levels. Conclusions: All mechanisms upregulating the expression of iodine transporters and changes in villous stroma and microvessel area in the placental labyrinth can promote iodide transfer from mother to fetus in iodine deficiency, especially before the onset of fetal thyroid function. Compensatory NIS protein regulation in the placenta against decreased iodine intake mainly occurs during translation and glycosylation modification after translation. Pendrin may be more important than NIS in the mediation of placental iodide transport.

Metabolomics evaluation of repeated administration of potassium iodide on adult male rats.

The long-lasting consequence of a new iodine thyroid blocking strategy (ITB) to be used in case of nuclear accident is evaluated in male Wistar rats using a metabolomics approach applied 30 days after ITB completion. The design used 1 mg/kg/day of KI over 8 days. Thyroid hormones remained unchanged, but there was a metabolic shift measured mainly in thyroid then in plasma and urine. In the thyroid, tyrosine metabolism associated to catecholamine metabolism was more clearly impacted than thyroid hormones pathway. It was accompanied by a peripheral metabolic shift including metabolic regulators, branched-chain amino acids, oxidant stress and inflammation-associated response. Our results suggested that iodide intake can impact gut microbiota metabolism, which was related to host metabolic regulations including in the thyroid. As there were no clear clinical signs of dysfunction or toxicity, we concluded that the measured metabolomics response to the new ITB strategy, especially in thyroid, is unlikely to reveal a pathological condition but a shift towards a new adaptive homeostatic state, called 'allostatic regulation'. The question now is whether or not the shift is permanent and if so at what cost for long-term health. We anticipate our data as a start point for further regulatory toxicity studies.

A novel procedure to assess the non-enzymatic hydrogen-peroxide antioxidant capacity of metabolites with high UV absorption.

Assays assessing non-enzymatic hydrogen peroxide antioxidant capacities are often hampered by the high UV absorption of the sample itself. This is a typical problem in studies using plant extracts with high polyphenol content. Our assay is based on comparing the 405 nm absorption of the product of potassium iodine and hydrogen peroxide in the presence and absence of a putative hydrogen peroxide reactive antioxidant. This method is free of interference with either hydrogen peroxide or antioxidant self-absorption and it is also suitable for high-throughput plate reader applications.

Effect of various sources and levels of iodine, as well as the kind of diet, on the performance of young laying hens, iodine accumulation in eggs, egg characteristics, and morphotic and biochemical indices in blood.

Young hens were fed over a period of 150 d with 2 kinds of diets including corn and soybean meal or corn, soybean, and rapeseed meal. Diets were enriched with potassium iodide (KI) or potassium iodate (KIO3) as an I source in amounts equal to 1, 3, or 5 mg of supplemented I/kg of feed. The hen performance, egg quality, hematological and morphotic indices in blood, hepatic enzyme activity, lipid indices in blood serum as well as I accumulation in wet egg content were determined. Introduction of 00-variety rapeseed meal into the diet improved the laying rate and feed conversion (P < 0.05); however, better egg weight was noted by feeding the hens with a diet without rapeseed meal. Use of KI as an I source enhanced the egg weight. The increased I level in the diet had an equivocal influence on egg weight, improved the feed conversion per 1 kg of eggs, and decreased the proportion of damaged eggs. The use of corn, soybean, and rapeseed meal in hen diets significantly improved yolk color; similar results were noted after an increase in I levels in the diets after 3 mo of feeding. Hematological indices of hen blood demonstrated significantly higher red blood cells numbers and hemoglobin concentrations with the use of KI. The use of a diet containing rapeseed meal led to an enhancement of hepatic enzyme activity, especially of alkaline phosphatase (P = 0.007). Lipid metabolism indices were not influenced by the kind of diet or the I source or level. The accumulation of I in wet egg content was negatively influenced by the use of a diet containing rapeseed meal (P = 0.000). The application of KI as an I source enhanced (P = 0.003) the accretion of I in eggs after 5 mo of treatment. Enhanced I supply significantly increased accumulation of I in eggs (P = 0.000) after 3 and 5 mo of the experiment from 260 and 310 to 1,011 and 1,256 µg/kg of wet egg content, respectively.

A new resonance Rayleigh scattering spectral method for determination of O3 with victoria blue B.

Ozone (O3) could be absorbed by boric acid-potassium iodide (BKI) absorbent solution to produce tri-iodine ion (I3(-)) that react with victoria blue B (VBB) to form the associated particle (VBB-I3)n and exhibited a strong resonance Rayleigh scattering (RRS) peak at 722 nm. Under the chosen conditions, the RRS peak intensity was linear with O3 concentration in the range of 0.2-50 µmol/L, with a linear regression equation of ΔI722=17.9c-45.4 and detection limit of 0.057 µmol/L. Accordingly, a simple, rapid and sensitive RRS spectral method was set up for determination of trace O3 in air, with satisfactory results.

Increased concentration of iodide in airway secretions is associated with reduced respiratory syncytial virus disease severity.

Recent studies have revealed that the human and nonrodent mammalian airway mucosa contains an oxidative host defense system. This three-component system consists of the hydrogen peroxide (H2O2)-producing enzymes dual oxidase (Duox)1 and Duox2, thiocyanate (SCN(-)), and secreted lactoperoxidase (LPO). The LPO-catalyzed reaction between H2O2 and SCN(-) yields the bactericidal hypothiocyanite (OSCN(-)) in airway surface liquid (ASL). Although SCN(-) is the physiological substrate of LPO, the Duox/LPO/halide system can generate hypoiodous acid when the iodide (I(-)) concentration is elevated in ASL. Because hypoiodous acid, but not OSCN(-), inactivates respiratory syncytial virus (RSV) in cell culture, we used a lamb model of RSV to test whether potassium iodide (KI) could enhance this system in vivo. Newborn lambs received KI by intragastric gavage or were left untreated before intratracheal inoculation of RSV. KI treatment led to a 10-fold increase in ASL I(-) concentration, and this I(-) concentration was approximately 30-fold higher than that measured in the serum. Also, expiratory effort, gross lung lesions, and pulmonary expression of an RSV antigen and IL-8 were reduced in the KI-treated lambs as compared with nontreated control lambs. Inhibition of LPO activity significantly increased lesions, RSV mRNA, and antigen. Similar experiments in 3-week-old lambs demonstrated that KI administration was associated with reduced gross lesions, decreased RSV titers in bronchoalveolar lavage fluid, and reduced RSV antigen expression. Overall, these data indicate that high-dose KI supplementation can be used in vivo to lessen the severity of RSV infections, potentially through the augmentation of mucosal oxidative defenses.

Naproxen intercalates with DNA and causes photocleavage through ROS generation.

Naproxen is an important non-steroidal anti-inflammatory drug with many pharmacological and biological properties. In this study, we have attempted to ascertain the mode of action and the mechanism of binding of naproxen to DNA. We have also demonstrated that, upon irradiation with white light, naproxen generates reactive oxygen species, causing DNA cleavage. Generation of reactive oxygen species from photo-irradiated naproxen as determined spectrophotometrically was found to lead to nicking of plasmid DNA as analyzed by agarose gel electrophoresis. Without photo-irradiation, naproxen binds to DNA and forms drug-DNA complexes as revealed by spectroscopic techniques. Several experiments such as determination of the effect of urea, iodide-induced quenching and a competitive binding assay with ethidium bromide showed that naproxen binds to DNA primarily in an intercalative manner. These observations were further supported by CD analysis, viscosity measurements and molecular docking. Using DNA as a template, fluorescence resonance energy transfer between naproxen and ethidium bromide was also observed, further strengthening the evidence for intercalative binding of naproxen with DNA.

Structural variability between starch granules in wild type and in ae high-amylose mutant maize kernels.

Starch granule structure within wild-type and ae high-amylose mutant maize kernels has been mapped in situ using light, electron and atomic force microscopy, and both Raman and infra-red spectroscopy. The population of wild-type starch granules is found to be homogenous. The ae mutant granule population is heterogeneous. Heterogeneity in chemical and physical structure is observed within individual granules, between granules within cells, and spatially within the kernel. The highest level of heterogeneity is observed in the region where starch is first deposited during kernel development. Light microscopy demonstrates structural diversity through use of potassium iodide/iodine staining and polarised microscopy. Electron and atomic force microscopy, and infra-red and Raman spectroscopy defined the nature of the structural changes within granules. The methodology provides novel information on the changes in starch structure resulting from kernel development.

The effects of iodine level and source on iodine carry-over in eggs and body tissues of laying hens.

In the presented study the effect of different iodine (I) levels and sources in hen feed on the iodine concentration of different tissues, blood serum, and eggs of laying hens was studied. For this purpose, two experiments were conducted with 30 laying hens each. In these experiments feed was enriched with KI and Ca(IO(3))(2), respectively, at 0 (Control), 0.25, 0.5, 2.5 and 5 mg I/kg feed, resulting a analysed iodine level from 0.44 to 4.20 mg/kg feed. After four weeks experimental feeding the iodine concentrations of thyroid glands, blood, meat, liver, abdominal fat and eggs were measured with inductively coupled plasma mass spectrometry. The experimental treatment did not affect hen performance. The iodine supplementation significantly increased the iodine concentration of eggs (144-1304 µg/kg), thyroid glands (3367-5975 µg/g), blood serum (16-67 µg/kg) and liver (13-43 µg/kg). Meat (about 14 µg I/kg) and abdominal fat (about 12 µg I/kg) were not significantly affected by iodine treatment. Comparative regression analyses showed that at a similar iodine intake, the supply via KI resulted in significantly higher iodine deposition into eggs than Ca(IO(3))(2). Due to the high carry-over of iodine into eggs, eggs may considerably contribute to the iodine supply of the consumers.

Protection of wood from microorganisms by laccase-catalyzed iodination.

In the present work, Norway spruce wood (Picea abies L.) was reacted with a commercial Trametes versicolor laccase in the presence of potassium iodide salt or the phenolic compounds thymol and isoeugenol to impart an antimicrobial property to the wood surface. In order to assess the efficacy of the wood treatment, a leaching of the iodinated and polymerized wood and two biotests including bacteria, a yeast, blue stain fungi, and wood decay fungi were performed. After laccase-catalyzed oxidation of the phenols, the antimicrobial effect was significantly reduced. In contrast, the enzymatic oxidation of iodide (I(-)) to iodine (I(2)) in the presence of wood led to an enhanced resistance of the wood surface against all microorganisms, even after exposure to leaching. The efficiency of the enzymatic wood iodination was comparable to that of a chemical wood preservative, VP 7/260a. The modification of the lignocellulose by the laccase-catalyzed iodination was assessed by the Fourier transform infrared spectroscopy-attenuated total reflectance (FTIR-ATR) technique. The intensities of the selected lignin-associated bands and carbohydrate reference bands were analyzed, and the results indicated a structural change in the lignin matrix. The results suggest that the laccase-catalyzed iodination of the wood surface presents an efficient and ecofriendly method for wood protection.

Enhancement of respiratory mucosal antiviral defenses by the oxidation of iodide.

Recent reports postulate that the dual oxidase (DUOX) proteins function as part of a multicomponent oxidative pathway used by the respiratory mucosa to kill bacteria. The other components include epithelial ion transporters, which mediate the secretion of the oxidizable anion thiocyanate (SCN(-)) into airway surface liquid, and lactoperoxidase (LPO), which catalyzes the H(2)O(2)-dependent oxidation of the pseudohalide SCN(-) to yield the antimicrobial molecule hypothiocyanite (OSCN(-)). We hypothesized that this oxidative host defense system is also active against respiratory viruses. We evaluated the activity of oxidized LPO substrates against encapsidated and enveloped viruses. When tested for antiviral properties, the LPO-dependent production of OSCN(-) did not inactivate adenovirus or respiratory syncytial virus (RSV). However, substituting SCN(-) with the alternative LPO substrate iodide (I(-)) resulted in a marked reduction of both adenovirus transduction and RSV titer. Importantly, well-differentiated primary airway epithelia generated sufficient H(2)O(2) to inactivate adenovirus or RSV when LPO and I(-) were supplied. The administration of a single dose of 130 mg of oral potassium iodide to human subjects increased serum I(-) concentrations, and resulted in the accumulation of I(-) in upper airway secretions. These results suggest that the LPO/I(-)/H(2)O(2) system can contribute to airway antiviral defenses. Furthermore, the delivery of I(-) to the airway mucosa may augment innate antiviral immunity.

6 Iodo-delta-lactone reproduces many but not all the effects of iodide.

BACKGROUND: Iodide has direct effects on thyroid function. Several iodinated lipids are biosynthesized by the thyroid and they were postulated as intermediaries in the action of iodide. Among them 6 iodo-delta-lactone (IL-delta) has been identified and proposed to play a role in thyroid autoregulation. The aim of this study was to compare the effect of iodide and IL-delta on several thyroid parameters. METHODS: Thyroid bovine follicles were incubated with the different compounds during three days. RESULTS: KI and IL-delta inhibited iodide uptake, total protein and Tg synthesis but only KI had an effect on NIS and Tg mRNAs levels. Both compounds inhibited Na+/K+ ATPase and deoxy-glucose uptake. As PAX 8, FOXE 1 and TITF1 are involved in the regulation of thyroid specific genes their mRNA levels were measured. While iodide inhibited the expression of the first two, the expression of TITF1 was stimulated by iodide and IL-delta had no effect on these parameters. CONCLUSION: These findings indicate that IL-delta reproduces some but not all the effects of excess iodide. These observations apply for higher micromolar concentrations of iodide while no such effects could be demonstrated at nanomolar iodide concentrations.

A new enzyme-catalytic resonance scattering assay for glucose in serum using cationic surfactant.

In acetic acid buffer solution, glucose oxidase (GOD) catalyzed the dissolved oxygen oxidation of glucose to form H(2)O(2). In succession, horseradish peroxidase (HRP) catalyzed the H(2)O(2) oxidizing excess I(-) to form I(3)(-). The I(3)(-) combined with a cationic surfactant (CS) such as tetradecyl dimethyl benzyl ammonium chloride (TDMBA) to produce TDMBA-I(3) association particles that exhibited the strongest resonance scattering (RS) peak at 460 nm. The enhanced RS intensity at 460 nm was linear with glucose concentration in the range of 2.0 x 10(-8)-2.0 x 10(-6) mol/L, with a detection limit of 8.5 x 10(-9) mol/L. The glucose in serum samples were assayed by the enzyme catalytic RS assay and by spectrophotometry. The results of both assays showed a close correlation. This assay has simplicity, sensitivity and good specificity for quantitative determination of glucose.

Saccharomyces cerevisiae: the effect of different forms and concentrations of iodine on uptake and yeast growth.

The essentiality of iodine for humans, especially in the early stages of life, is well recognized. The chemical forms of iodine in food supplements, infant formulae and iodated salt are either iodide (KI) or iodate (KIO(3)). Because there are no or rare data about iodine uptake by yeasts, we investigated the influence of different sources of iodine, as KI, KIO(3) and periodate (KIO(4)), on its uptake in and growth of the model yeast Saccharomyces cerevisiae. KIO(3) inhibited the growth of the yeast the most and already at a 400 microM initial concentration in the growth medium; the OD was reduced by 23% in comparison with the control, where no KIO(3) was added. The uptake of different iodine sources by the yeast S. cerevisiae was minimal, in total <1%. Tracer experiments with radioactive (131)I added as KI showed that the yeast S. cerevisiae does not have the ability to transform KI into volatile species. We investigated the specificity of iodine uptake added as KIO(3) in the presence of Na(2)SeO(4) or ZnCl(2) or K(2)CrO(4) in the growth medium, and it was found that chromate had the most influence on reduction of KIO(3) uptake.

Effect of charge distribution in a flexible loop on the bioluminescence color of firefly luciferases.

Firefly luciferase is a monooxygenase that catalyzes the ATP-dependent conversion of firefly luciferin into a luciferyl-adenylate, which is oxidized to an electronically excited oxyluciferin in a multistep reaction and produces visible light with a remarkable quantum yield. The bioluminescence color of firefly luciferases is determined by the luciferase structure and assay conditions. Among different beetles, only luciferase from Phrixotrix railroad worm (Ph(RE)) emits red bioluminescence, naturally. The presence of Arg353 in Ph(RE) luciferase, which corresponds to the deleted residue in the other luciferases, is an important distinctive structural feature of it. Insertion of Arg356 into a green-emitter luciferase (Lampyris turkestanicus), corresponding to Arg353 in Phrixotrix hirtus, changed the emitted light from green to red. To further clarify the effect of this position on the light shift mechanism, four residues with similar sizes but different charges (Arg, Lys, Glu, and Gln) were inserted into Photinus pyralis luciferase, using site-specific insertion mutagenesis. Insertion of a residue with a positive side chain (Arg356 and Lys356) changed the light color to red, while insertion of a residue with a negative side chain (Glu356) had little effect on color. Insertion of a neutral residue (Gln356) at this position was performed without any change in bioluminescence spectra. Insertion of positively charged residues in this loop took place with a series of structural changes which were confirmed by fluorescence spectroscopy and homology modeling. Homology modeling reveals the appearance of a bulge in a flexible loop (T352-P359) upon mutation which shifts to the left side with a color change from green to red.

Thyroid dose estimation with potassium iodide (KI) administration in a nuclear emergency.

In a breach-of-containment nuclear reactor accident, the near-field exposure is primarily through inhalation of radioiodine. Thyroid blockade by oral potassium iodide (KI) is a practical and effective protective measure for the general public in such an emergency. The retention functions incorporating the thyroid blocking effects by KI have been derived using a standard three-compartment model of iodine metabolism. This allows more accurate estimation of the thyroid dose by calculating the blocking factor.

Uso racional del yoduro potásico durante el embarazo y la lactancia / Rational use of potassium iodine during pregnancy and lactation

La deficiencia de yodo grave y moderada durante la gestación y la lactancia afecta a la función tiroidea de la madre y del neonato, así como al desarrollo neuropsicológico del niño. En la mayoría de los países de Europa el estado nutricional de yodo de gestantes y recién nacidos es deficiente, pues las necesidades de yodo en las mujeres embarazadas y en las que amamantan a sus hijos son al menos el doble de las recomendadas para la población adulta. Estudios realizados en España confirman que la mayoría de las mujeres se encuentran en yododeficiencia durante la gestación y la lactancia y muestran quela ingesta de sal yodada no es suficiente para conseguir un estado nutricional de yodo óptimo en esta población. Varios estudios demuestran el efecto beneficioso de la suplementación de yodo de las embarazadas sobre el estado nutricional de yodo y la función tiroidea de la madre y el recién nacido. Las mujeres embarazadas, las que amamantan a sus hijos y las que planifican su gestación deberían recibir suplementos de yodo en forma de tabletas de yoduro potásico (AU)

Severe and mild iodine deficiency during pregnancy and lactation affects thyroid function of the mother and neonate as well as the infant’s neuropsychological development. In most European countries, maternal and neonatal iodine status is deficient, since iodine requirements in pregnant and lactating women are almost double those recommended for non pregnantor non-lactating women. Studies performed in Spain confirm that most women are iodine deficient during pregnancy and lactation, and show that iodized salt intake is insufficient to achieve optimal iodine nutritional status in this population. Several studies have demonstrated the beneficial effect of iodine supplementation in pregnant women on the iodine and thyroid status of both mother and neonate. Pregnant and breastfeeding women and women planning to become pregnant should take iodine supplements in the form of potassium iodide tablets (AU)