Delta-aminolevulinic Acid Dehydratase Research Articles

Exogenous nitric oxide requires endogenous hydrogen sulfide to induce the resilience through sulfur assimilation in tomato seedlings under hexavalent chromium toxicity

Nitric oxide (NO) and hydrogen sulfide (H2S), versatile signaling molecules, play multiple roles in plant growth, physiological and biochemical processes under heavy metal stress. However, the mechanisms through which NO in association with endogenous H2S mediated hexavalent chromium Cr(VI) toxicity mitigation are still not fully understood. Therefore, we investigated the role of NO and H2S in sulfur (S)-assimilation and the effect of NO on endogenous H2S, and cysteine (Cys) biosynthesis and maintenance of cellular glutathione (GSH) pool in tomato seedlings under Cr(VI) stress. Cr(VI) toxicity caused an increase in reactive oxygen species (ROS; O2•– and H2O2) formation and activity of chlorophyll (Chl) degrading enzyme [Chlorophyllase (Chlase)] and decrease in seedlings growth attributes, Chl a and b content, and activity of Chl synthesizing enzyme [δ-aminolevulinic acid dehydratase (δ-ALAD)], gas exchange parameters, S-assimilation, and Cys and H2S metabolism. An increase in the content of glycinebetaine (GB), total soluble carbohydrates (TSCs) and total phenols (TPls), and decrease in DNA damage and ROS in NO treated seedlings conferred Cr(VI) toxicity tolerance. Under Cr(VI) toxicity conditions, the inclusion of H2S scavenger hypotaurine (HT) in growth medium containing NO validated the role of endogenous H2S in S-assimilation, H2S and Cys and GSH metabolism by withdrawing activity of enzymes involved in S-assimilation [adenosine 5-phosphosulfatereductase (APS-R), ATP-sulfurylase (ATP-S)], in the biosynthesis of H2S [L-cysteine desulfhydrase (L-CD) and D-cysteine desulfhydrase (D-CD)], Cys [O-acetylserin (thiol) lyase (OAST-L)], and GSH [glutamylcysteine synthetase (γ-GCS) and glutathione synthetase (GS)], and in antioxidant system. On the other hand, application of cPTIO [2-(4–32 carboxyphenyl)-4,4,5,5-tetramethylimidazoline-1-oxyl-3-oxide], a NO scavenger and HT diminished the effect of NO on internal H2S levels, Cys and glutathione homeostasis, and S-assimilation, which resulted in poor immunity against oxidative stress induced by Cr(VI) toxicity. The obtained results postulate that NO-induced internal H2S conferred tolerance of tomato seedlings to Cr(VI) toxicity and maintained better photosynthesis process and plant growth.

Mercury toxicity in pregnant and lactating rats: zinc and N-acetylcysteine as alternative of prevention.

This study evaluated the toxic effects of inorganic mercury (Hg) in pregnant and lactating rats, as well as the possible protective effect of zinc (Zn) and N-acetylcysteine (NAC). Pregnant and lactating rats were pre-treated with ZnCl2 (27mg/kg) and/or NAC (5mg/kg) and after 24h, they were exposed to HgCl2 (10mg/kg). Animals were sacrificed 24h after Hg exposure, and biochemical tests and metal determination were performed. Regarding pregnant rats, Hg exposure caused kidney, blood, and placenta δ-aminolevulinic acid dehydratase (δ-ALA-D) activity inhibition, and the pre-treatments showed a tendency of protection. Moreover, all the animals exposed to Hg presented high Hg levels in the kidney, liver, and placenta when compared with control group. Pregnant rats pre-exposed to Zn (Zn-Hg and Zn/NAC-Hg groups) presented an increase in hepatic metallothionein levels. Therefore, lactating rats exposed to Hg presented renal and blood δ-ALA-D inhibition; the pre-treatments showed a tendency to prevent the renal δ-ALA-D inhibition and prevented the blood δ-ALA-D inhibition caused by Hg. Lactating rats exposed to Hg presented high Hg levels in the kidney and liver. These results showed that 10mg/kg of HgCl2 causes biochemistry alterations in pregnant and lactating rats, and Zn and NAC present promising results against these damages.

Sub-chronic administration of flavonoid fraction Daflon improve lead-induced alterations in delta-aminolevulinic acid dehydratase activity, erythrocytic parameters, and erythrocyte osmotic fragility in Wistar rats

This study investigated the ameliorative potential of micronized purified flavonoid fraction Daflon on lead-induced alterations in heme-biosynthetic enzyme delta-aminolevulinic acid dehydratase (δ-ALAD) activity and erythrocytic parameters. Thirty-two male adult Wistar rats were divided by simple randomization into four groups of eight rats each. Lead acetate was administered orally at a dose of 190 mg/kg (1/20th of the LD50) body weight for 42 days, either alone or with Daflon at a dose of 100 mg/kg body weight. Blood lead level (BLL) was analyzed using the microwave plasma atomic emission spectrometer (MP-AES), while δ-ALAD activity was quantified using assay kit. Erythrocyte parameters (RBC, packed cell volume, Hb, MCV, and MCHC) were assayed using an automated hematologic analyzer. Erythrocyte osmotic fragility (EOF) was also analyzed using standard procedure. Sub-chronic exposure to lead acetate caused impairment in δ-ALAD activity, erythrocytic parameters, and increased EOF. Conversely, Daflon improved the activity of δ-ALAD and erythrocytic parameters by protecting the integrity of the erythrocyte membrane. The results of our study revealed that sub-chronic exposure of rats to lead acetate induced impairment in δ-ALAD, erythrocytic parameters, and EOF, while pretreatment with Daflon alleviated the ensued changes, and therefore suggested that dietary supplementation with Daflon-enriched foods may be a promising prophylactic agent for individuals living in lead toxicity prone environments.

Intercorrelation mechanism of Hypoxia and Cordycepin Biosynthesis in Zombie fungus

Cordycepin is an anticancer metabolite produces by a zombie fungus species of Cordyceps militaris. They are capable to infect and hijack insect’s nervous neuron system. Hypoxic environment commonly must be faced by the pathogenic fungus during infection either this zombie fungus. They activate oxygen sensing mode, heme, siderophore, and sterol biosynthesis to overcome it. Underlined our previous study that liquid surfaced culture of C. militaris NBRC103752 produced a higher amount of cordycepin than submerged culture, suggesting that hypoxic conditions might induce it. However, when and how the mechanism of cordycepin production started in liquid surfaced culture is not understood, yet. In our present study, the combination of transcriptomics and gas chromatography-mass spectrometry were carried out during the production phases of cordycepin (5d, 12d, and 19d of incubation periods) and the mechanism of cordycepin production was figured out. The expression of genes in the fermentation pathway and the oxidative phosphorylation pathway were significantly upregulated and down regulated, respectively. Expression of four genes in the heme biosynthesis, including 5-aminolevulinic acid synthase (CCM_01504), delta-aminolevulinic acid dehydratase (CCM_00935), coproporphyrinogen III oxidase (CCM_07483) and cytochrome c oxidase15 (CCM_05057) were upregulated at the beginning of the exponential phase (12d). Further, the activation of Zn(2)-C6 transcription factor that regulates the iron acquisition and ergosterol biosynthesis significantly upregulated and a metabolite reporter adenosine was detected only at 12d. The results in the present study show the correlation between hypoxia and the accumulation of heme before cordycepin biosynthesis.

Relationship Between Lead (Pb) Concentration in Soil, Grass , Blood, Milk and δ-aminolevulinic Acid Dehydratase (ALAD) Activity , Hemoglobin (Hb) and Hematocrit (Hct) in Grazing Cows from Vicinity of Smelter “Trepça” in Kosovo

Relationship Between Lead (Pb) Concentration in Soil, Grass , Blood, Milk and δ-aminolevulinic Acid Dehydratase (ALAD) Activity , Hemoglobin (Hb) and Hematocrit (Hct) in Grazing Cows from Vicinity of Smelter “Trepça” in Kosovo

Evaluation of antioxidant activity and toxicity of sulfur- or selenium-containing 4-(arylchalcogenyl)-1H-pyrazoles.

Pyrazoles represent a significant class of heterocyclic compounds that exhibit pharmacological properties. The present study aimed to investigate the antioxidant potential of pyrazol derivative compounds in brain of mice in vitro and the effect of pyrazol derivative compounds in the oxidative damage and toxicity parameters in mouse brain and plasma of mice. The compounds tested were 3,5-dimethyl-1-phenyl-4-(phenylselanyl)-1H-pyrazol (1a), 3,5-dimethyl-4-(phenylselanyl)-1H-pyrazole (2a), 4-((4-methoxyphenyl)selanyl)-3,5-dimethyl-1-phenyl-1H-pyrazole (3a), 4-((4-chlorophenyl)selanyl)-3,5-dimethyl-1-phenyl-1H-pyrazole (4a), 3,5-dimethyl-1-phenyl-4-(phenylthio)-1H-pyrazole (1b), 3,5-dimethyl-4-(phenylthio)-1H-pyrazole (2b), 4-((4-methoxyphenyl)thio)-3,5-dimethyl-1-phenyl-1H-pyrazole (3b), 4-((4-chlorophenyl)thio)-3,5-dimethyl-1-phenyl-1H-pyrazole (4b), and 3,5-dimethyl-1-phenyl-1H-pyrazole (1c). In vitro, 4-(arylcalcogenyl)-1H-pyrazoles, at low molecular range, reduced lipid peroxidation and reactive species in mouse brain homogenates. The compounds also presented ferric-reducing ability as well nitric oxide-scavenging activity. Especially compounds 1a, 1b, and 1c presented efficiency to 1,1-diphenyl-2-picryl-hydrazyl-scavenging activity. Compounds 1b and 1c presented 2,20 -azino-bis(3-ethylbenzthiazoline-6-sulfonic acid)-scavenging activity. In vivo assays demonstrated that compounds 1a, 1b, and 1c (300 mg/kg, intragastric, a single administration) did not cause alteration in the of δ-aminolevulinic acid dehydratase activity, an enzyme that exhibits high sensibility to prooxidants situations, in the brain, liver, and kidney of mice. Compound 1c reduced per se the lipid peroxidation in liver and brain of mice. Toxicological assays demonstrate that compounds 1a, 1b, and 1c did not present toxicity in the aspartate aminotransferase, alanine aminotransferase, urea, and creatinine levels in the plasma. In conclusion, the results demonstrated the antioxidant action of pyrazol derivative compounds in in vitro assays. Furthermore, the results showed low toxicity of compounds in in vivo assays.

Blood lead level (BLL), δ-aminolevulinic acid dehydratase activity (ALAD), hemoglobin (Hb) and hematocrit (hct) in primary school-children and adult residents living in smelter rural areas in Kosovo

The objective of this study was to investigate the effects of chronic environmental lead (Pb) exposure in blood lead level (BLL), δ-aminolevulinic acid dehydratase (ALAD) activity, hemoglobin (Hb) amount and hematocrit (Hct) value in primary schoolchildren and adults. Blood was obtained for BLL, ALAD, Hb and Hct measurements in 23 primary schoolchildren (girls and boys) and 117 adult residents (women and men) living in three villages (Kelmend, Boletin and Zhazhë) defined by concentric circles 2, 3 and 5 km in radius drawn around from the smelter-refinery complex “Trepça”in Zveçan and in Koliq village 40 km away. As expected, BLLs were substantially higher in the schoolchildren from smelter area compared with control (11 ± 4.2 µg/L and 6.9 ± 1.6 µg/L respectively) and in adult residents from Kelmend, Boletin and Zhazhë (24 ± 11.8, 12 ± 4.5, 11 ± 5.4 and 8.0 ± 2.8 µg/L respectively). Blood ALAD activity of children in Zhazhë is 16% inhibited compared to control and blood ALAD activity in adults in villages from smelter area is 32, 3%, 48, 4% and 17, 8% inhibited compared to control. There is no difference of Hb and Hct values in schoolchildren from Zhazhë and in adult residents from Kelmend and Zhazhë compared with control. Results of this study provide evidence of moderate inverse correlation between BLL and ALAD activity in both examined cohorts from smelter area. The inhibition of ALAD activity in primary schoolchildren and adults occurred at blood lead levels < 24 µg/L; consequently it can cause an increase of δ- Aminolevulinic acid.

Exogenous melatonin mitigates boron toxicity in wheat.

Boron (B) toxicity is an important abiotic constraint that limits crop productivity mainly in arid and semi-arid areas of the world. High levels of B in soil disturbs several physiological and biochemical processes in plant. The aim of this study was to investigate the function of melatonin (Mel) in the regulation of carbohydrate and proline (Pro) metabolism, photosynthesis process and antioxidant system of wheat seedlings under B toxicity conditions. High levels of B inhibited net photosynthetic rate (PN), stomatal conductance (gs), content of chlorophyll (Chl) a, b, δ-aminolevulinic acid (δ-ALA), nitrogen (N) and phosphorus (P), and increased accumulation of B, Chl degradation and activity of chlorophyllase (Chlase; a Chl degrading enzyme), and downregulated the activity of enzymes (δ-ALAD; δ-aminolevulinic acid dehydratase) involved in the biosynthesis of photosynthesis pigments, photosynthesis (carbonic anhydrase and ribulose-1,5-bisphosphate carboxylase/oxygenase) and carbohydrate metabolism (cell wall invertase, CWI) in wheat seedlings. Also, high levels of B caused oxidative damage by increasing the content of malondialdehyde, superoxide anion and H2O2, and activity of glycolate oxidase (an H2O2-producing enzyme) in leaves of seedlings. However, foliar application of Mel significantly improved photosynthetic pigments concentration by increasing δ-ALA, δ-ALAD and decreasing Chl degradation and Chlase activity and led to an increase of plant growth attributes under both B toxicity and non-toxicity conditions. Under normal and B toxicity conditions, exogenous Mel also improved content of N, P, total soluble carbohydrates (TSCs) and Pro, and upregulated activity of CWI and Δ1-pyrroline-5-carboxylate synthetase. Mel significantly suppressed the adverse effects of excess B by alleviating cellular oxidative damage through enhanced reactive oxygen species scavenging by superoxide dismutase, catalase, ascorbate peroxidase, glutathione reductase and lipoxygenase, and content of total phenolic compounds (TPC), ascorbate and reduced glutathione. These results postulate that Mel induced plant defense mechanisms by enhancing Pro, TSCs, TPC, nutrients (N and P) uptake and enzymatic and non-enzymatic antioxidants.

The Se…S/N interactions as a possible mechanism of δ-aminolevulinic acid dehydratase enzyme inhibition by organoselenium compounds: A computational study

Organoselenium compounds present many pharmacological properties and are promising drugs. However, toxicological effects associated with inhibition of thiol-containing enzymes, such as the δ-aminolevulinic acid dehydratase (δ-AlaD), have been described. The molecular mechanism(s) by which they inhibit thiol-containing enzymes at the atomic level, is still not well known. The use of computational methods to understand the physical–chemical properties and biological activity of chemicals is essential to the rational design of new drugs. In this work, we propose an in silico study to understand the δ-AlaD inhibition mechanism by diphenyl diselenide (DPDS) and its putative metabolite, phenylseleninic acid (PSA), using δ-AlaD enzymes from Homo sapiens (Hsδ-AlaD), Drosophila melanogaster (Dmδ-AlaD) and Cucumis sativus (Csδ-AlaD). Protein modeling homology, molecular docking, and DFT calculations are combined in this study. According to the molecular docking, DPDS and PSA might bind in the Hsδ-AlaD and Dmδ-AlaD active sites interacting with the cysteine residues by Se…S interactions. On the other hand, the DPDS does not access the active site of the Csδ-AlaD (a non-thiol protein), while the PSA interacts with the amino acids residues from the active site, such as the Lys291. These interactions might lead to the formation of a covalent bond, and consequently, to the enzyme inhibition. In fact, DFT calculations (mPW1PW91/def2TZVP) demonstrated that the selenylamide bond formation is energetically favored. The in silico data showed here are in accordance with previous experimental studies, and help us to understand the reactivity and biological activity of organoselenium compounds.

Blood lead levels and aberrant DNA methylation of the ALAD and p16 gene promoters in children exposed to environmental-lead

BackgroundLead (Pb) is a well-known toxic heavy metal which can have serious public health hazards. As of today, there is no safe threshold for Pb exposure, especially for children. Lead exposure has been associated with adverse health outcomes involving epigenetic mechanisms, such as aberrant DNA methylation. The objective of the present study was to elucidate the associations between blood lead levels (BLLs) and gene-specific promoter DNA methylation status in environmental Pb-exposed children from Kabwe, Zambia. MethodsA cross-sectional study was conducted using 2 to 10-year-old children from high Pb exposed area (N = 102) and low Pb exposed area (N = 38). We measured BLLs using a LeadCare II analyzer and investigated the methylation status of the ALAD and p16 gene promoters by methylation–specific PCR. ResultsThe mean BLLs were 23.7 μg/dL and 7.9 μg/dL in high Pb exposed and low Pb exposed children, respectively. Pb exposure was correlated with increased methylation of the ALAD and p16 genes. The promoter methylation rates of ALAD and p16 in high Pb exposed children were 84.3% and 67.7%, and 42.1% and 44.7% in low Pb exposed children, respectively. Significantly increased methylation was found in both genes in high Pb exposed children compared with low Pb exposed children (p < 0.05). Children with methylated ALAD and p16 genes showed an increased risk of Pb poisoning (odd ratio >1) compared to the unmethylated status. ConclusionsThis study for the first time tries to correlate promoter methylation status of the ALAD and p16 genes in environmental Pb-exposed children from Kabwe, Zambia as a representative. The result suggests that Pb exposure increases aberrations in ALAD and p16 gene methylation, which may be involved in the mechanism of Pb toxicity.

Melatonin and calcium function synergistically to promote the resilience through ROS metabolism under arsenic-induced stress

The interplay between melatonin (Mel) and calcium (Ca2+) in enhancing tolerance to metalloid toxicity and underlying physiological and biochemical mechanisms of this relationship still remains unknown. The present study reveals that the signaling molecules Mel and/or Ca2+ enhanced tolerance of Vicia faba (cv. Tara) plant to metalloid arsenic (As) toxicity. However, a combination of Mel and Ca2+ was more efficient than alone. Plants grew with As exhibited enhanced hydrogen peroxide, superoxide anion, electrolyte leakage, lipid peroxidation together with increased reactive oxygen species (ROS) producing enzymes, such as NADPH oxidase and glycolate oxidase (GOX). On the contrary, an inhibition in chlorophyll (Chl) biosynthesis and gas exchange parameters (net photosynthetic rate, stomatal conductance, intercellular carbon dioxide concentration) was observed. Under As toxicity conditions, the application of Mel and Ca2+ synergistically suppressed the plants’ program cell death features (nucleus condensation and nucleus fragmentation) in guard cells of stomata, DNA damage, and formation of ROS in guard cells, leaves and roots. Moreover, it enhanced gas exchange parameters and activity of enzymes involved in photosynthesis process (carbonic anhydrase and RuBisco), Chl biosynthesis (δ-aminolevulinic acid dehydratase), and decreased activity of Chl degrading enzyme (chlorophyllase) under As toxicity conditions. Our investigation evidently established that expression of ATP synthase, Ca2+-ATPase, Ca2+-DPKase, Hsp17.6 and Hsp40 was found maximum in the plants treated with Mel + Ca2+, resulting in higher tolerance of plants to As stress. Also, increased total soluble carbohydrates, cysteine, and Pro accumulation with increased Pro synthesizing enzyme (Δ1-pyrroline-5-carboxylate synthetase (P5CS) and decreased Pro degrading enzyme (proline dehydrogenase) in Mel + Ca2+ treated plants conferred As toxicity tolerance. The obtained results postulate strong evidence that the application of Mel along with Ca2+ enhances resilience against As toxicity by upregulating the activity of plasma membrane H+-ATPase, enzymes involved in antioxidant system, and ascorbate-glutathione pathway.

Effects of iron deficiency and exogenous sucrose on the intermediates of chlorophyll biosynthesis inMalushalliana.

Malus halliana is an iron (Fe)-efficient apple rootstock growing in calcareous soil that shows obvious 'greenness' traits during Fe deficiency. Recent studies have shown that exogenous sugars can be involved in abiotic stress. To identify the key regulatory steps of chlorophyll (Chl) biosynthesis in M. halliana under Fe deficiency and to verify whether exogenous sucrose (Suc) is involved in Fe deficiency stress, we determined the contents of the Chl precursor and the expression of several Chl biosynthetic genes in M. halliana. The results showed that Fe deficiency caused a significant increase in the contents of protoporphyrin IX (Proto IX), Mg-protoporphyrin IX (Mg-Proto IX) and protochlorophyllide (Pchlide) in M. halliana compared to the Fe-sensitive rootstock Malus hupehensis. Quantitative real-time PCR (RT-qPCR) also showed that the expression of protoporphyrinogen oxidase (PPOX), which synthesizes Proto IX, was upregulated in M. halliana and downregulated in M. hupehensis under Fe deficiency. Exogenous Suc application prominently enhanced the contents of porphobilinogen (PBG) and the subsequent precursor, whereas it decreased the level of δ-aminolaevulinic acid (ALA), suggesting that the transformation from ALA to PBG was catalyzed in M. halliana. Additionally, the transcript level of δ-aminolevulinate acid dehydratase (ALAD) was noticeably upregulated after exogenous Suc treatment. This result, combined with the precursor contents, indicated that Suc accelerated the steps of Chl biosynthesis by modulating the ALAD gene. Therefore, we conclude that PPOX is the key regulatory gene of M. halliana in response to Fe deficiency. Exogenous Suc enhances M. halliana tolerance to Fe deficiency stress by regulating Chl biosynthesis.

Melatonin and Gibberellic Acid Promote Growth and Chlorophyll Biosynthesis by Regulating Antioxidant and Methylglyoxal Detoxification System in Tomato Seedlings Under Salinity

The beneficial roles of melatonin (Mel) and gibberellic acid (GA3) in the biosynthesis of photosynthetic pigments, osmoregulation, and methylglyoxal (MG) detoxification and antioxidant system were studied in tomato (Solanum lycopersicum L. cv. Five Star) seedlings under NaCl stress. The exogenous application of Mel (100 µM) and GA3 (1.4 µM) together more efficiently affected growth performance of seedlings under salt stress. The decreased chlorophyll (Chl) degradation and Chl-degrading enzyme (chlorophyllase) activity in seedlings receiving Mel plus GA3 resulted in increased Chl content by upregulating Chl synthesizing enzyme (δ-aminolevulinic acid dehydratase) under salinity. Exogenous Mel plus GA3 suppressed the overproduction of reactive oxygen species (ROS; superoxide and hydrogen peroxide) and activity of glycolate oxidase. Application of Mel with GA3 reduced MG content by enhancing the activity of enzymes (glyoxalase I and glyoxalase II) involved in the MG detoxification system. Both Mel and GA3 together protected seedlings from ROS induced damage by regulating Δ1-pyrroline-5-carboxylate synthetase activity, and content of proline (Pro) and glycine betaine (GB). Seedlings receiving Mel + GA3 exhibited a substantial upregulated activity of catalase, ascorbate peroxidase, glutathione reductase, dehydroascorbate reductase, monodehydroascorbate reductase, glutathione peroxidase, polyphenol oxidase and lipoxygenase, and redox homeostasis that reduced oxidative damage induced by salinity. These outcomes advocate that Mel and GA3 played beneficial roles in Chl, Pro and GB biosynthesis, and improved redox homeostasis, and MG detoxification and antioxidant system under salt stress.

Multimeric Proteins that Can Come Apart, Change Shape, and Reassemble Differently with Functional Consequences ‐ Morpheeins

Herein we describe progress in a class of shape‐shifting proteins originally called morpheeins. These are homomultimers that can dissociate, change conformation in the dissociated state, and reassemble into a structurally and functionally distinct multimer. The original observation was of an octamer/dimer/dimer*/hexamer equilibrium wherein the conformational change was a hinge motion between folded domains of porphobilinogen synthase (PBGS). This phenomenon is established as a mechanism for allosteric regulation, druggable, and the structural basis for an inborn error of metabolism.Many aspects of the original observations in the early 2000s were unexpected and peculiar; suggesting a structural metastability. At the time, vocabulary to describe such multi‐structural and multi‐functional protein dynamics were limited. We defined morpheeins as (homo‐multimeric) proteins that can come apart, change shape and reassemble differently with functional consequences. A Wikipedia page introduced a dice model of a morpheein (see image), and showed how multimer‐specific surface cavities can provide a general mechanism for drug action. In the beginning, multimeric proteins with more than one shape (in the absence of refolding) strongly defied the one sequence, one structure, one function paradigm. Two decades later, there are a growing number of examples of multimeric proteins with multiple structures and other proteins with multiple functions, many in the absence of any chemical modification or significant refolding.The quaternary structure dynamics of PBGS have now been extensively described. Interestingly, one consequence of our focus on PBGS is that the literature began equated the specific properties of PBGS with all of the factors that must be in place for a protein to be classified as a morpheein. This is akin to saying that in order to be considered a mammal, an organism must have all of the characteristics of a rat. Herein, we set straight the definition of a morpheein; it is a protein that can come apart, change conformation in the dissociated state, and reassemble to a structurally and functionally distinct multimer. Alternate assemblies need not have different stoichiometries. Assembly interchange need not be triggered by ligand binding or catalysis. Functional difference need not be high activity and low activity (as in allosteric regulation of enzyme action), but can be moonlighting activities. Often the conformational change is a repositioning of folded domains relative to each other.An excellent example of a moonlighting protein that appears to be a morpheein, and has also been called a transformer, is the VP40 protein of the Ebola virus. It too exists as dimers or hexamer or octamer and each alternate assembly is responsible for an alternate function in the viral life cycle. The individual assemblies are stabilized through interaction with a “third party”. A third likely example is the family of alternate assemblies of ribonucleotide reductase, whose timely interchange amongst alternate assemblies appears essential to the regulation of nucleotide pools. A fourth example is HIV integrase, whose three domains reposition relative to each other as part of viral integration. These examples will be illustrated.Support or Funding InformationNIH grants 5R01 NS100081 and P30 CA006927Dice image depicting an interchange of morpheein forms and a ligand that stabilizes one form.Figure 1

Lead toxicity on a sentinel species subpopulation inhabiting mangroves with different status conservation

Lead is a priority pollutant introduced in the aquatic environment by different sources commonly located in estuarine regions, such as ports, marinas and industries. Environmental agencies around the world set the maximum allowable concentration of lead in effluents, surface water and sediment, but few studies reported its accumulation and chronic toxicity in mangrove benthic invertebrates using concentrations believed to be safe. In the case of Brazilian mangrove environments, Ucides cordatus is a crab species of choice to be used in bioaccumulation studies. We have assessed biomarkers’ responses (DNA strand breaks, micronucleated cells, metallothioneins, enzymatic activity of aminolevulinic acid dehydratase and neutral red retention time) and the total bioaccumulation in six tissues of U. cordatus crabs resident to mangrove areas under different conservation status during a 28-day period bioassay. We also investigated Pb subcellular partition and biomarkers’ responses using a supposedly safe concentration (10 μg L−1). During the Pb exposure, the highest concentration of Pb was observed in crab gills. Crabs also showed a high ability to allocate Pb in detoxified forms. Multivariate analysis pointed out that bioaccumulation (total, active and detoxified) is linked to biomarkers. Even in supposedly safe dosage, U. cordatus triggered its defense mechanisms expressing more metallothioneins and presented relevant cyto-genotoxic damage. Our data suggest the development of biological tolerance to Pb in crabs from polluted areas. Our results provided a new insight about lead toxicity even at concentrations considered environmentally safe, which could support new strategies to manage estuarine areas considering their respective conservation status.

Diphenyl diselenide is as effective as Ebselen in a juvenile rat model of cisplatin-induced nephrotoxicity

BackgroundCisplatin (CIS) is widely used in the chemotreatment of pediatric tumors. However, the CIS use is limited because of its high incidence of toxicity, mainly nephrotoxicity. Although there are many studies about CIS-related nephrotoxicity in animal models, only a few studies focus on juvenile animals. Because redox disturbances have been associated with kidney damage induced by CIS, this study aimed to compare the effectiveness of Ebselen and diphenyl diselenide (PhSe)2 against nephrotoxicity induced by CIS in juvenile rats. MethodsJuvenile Wistar rats were randomly divided into six groups: rats from groups I to III received an intraperitoneal (i.p.) injection with saline solution. The other groups received CIS (i.p., 6 mg/kg) on the first day. One hour before CIS injection and on the next four days, animals of groups III and V were intragastrically treated with Ebselen (11 mg/kg) whereas those from groups IV and VI received (PhSe)2 (12 mg/kg). After 24 h of the last treatment, blood and kidney were collected, and the parameters of renal function and oxidative stress were determined. ResultsKidney damage induced by CIS was confirmed by the increase of creatinine, urea and uric acid levels in the blood of juvenile rats. The renal oxidative disturbance was characterized by an increase in the levels of thiobarbituric acid reactive substances (TBARS), protein carbonyl, and nitrogen oxides (Nox), as well as the decrease in non-protein thiol content (NPSH), glutathione-S-transferase (GST), catalase (CAT) and superoxide dismutase (SOD) activities. CIS inhibited the activities of δ-aminolevulinic acid dehydratase (δ-ALA-D) and Na+, K+-ATPase and down-regulated the Nrf2/Keap-1/HO-1 pathway in the kidney of juvenile rats. ConclusionBoth Ebselen and (PhSe)2 modulated back to the normal levels all parameters altered by the CIS administration in the kidney of juvenile rats. Thus, this study shows that (PhSe)2 was as effective as Ebselen in protecting the kidney against oxidative damage caused by CIS in rats.

Fundamentals of 5‐aminolevulinic acid photodynamic therapy and diagnosis: An overview

Abstract5‐Aminolevulinic acid‐based photodynamic therapy (ALA‐PDT) comprises photochemical and biological reactions, finally generating tumor destruction. Tumor selectivity of ALA‐PDT relies on the disturbed heme synthesis pathway in neoplastic cells. Tumor pretreatment with ALA induces two reactions: the first, self‐activation of the key enzyme porphobilinogen synthase and then protoporphyrin (PpIX) synthesis at low Fe++ availability. We emphasize that the toxic mechanism of ALA‐PDT triggered by 1O2 targets intracellular organelle destruction death pathways. PpIX fluorescence of tumors, particularly in glioblastoma, is used clinically for fluorescence guidance during neurosurgery. Targeting residual and metastatic cancer stem cells is a crucial challenge of ALA‐PDT, plausibly combined with other therapeutic modalities to encounter metastatic recurrences. We overview the different aspects of ALA‐PDT mechanism and future research and prospects.

Risk assessment of heavy metal toxicity by sensitive biomarker δ-aminolevulinic acid dehydratase (ALA-D) for onion plants cultivated in polluted areas in Kosovo

Biomarkers allow an integrated risk assessment of heavy metal pollution effects in living organisms. In this study, the biochemical effects of Cd, Cr, Ni, Pb and Zn pollution in agricultural soil and their accumulation in Alium cepa L. (onion) were evaluated with ALA-D enzyme response as a biomarker, along with δ-aminolevulinic acid (ALA) and total chlorophyll contents in leaves of this plant. Soil samples were randomly selected from agricultural areas in two regions, Mitrovica and Obiliqi, which are considered the most industrially polluted regions in Kosovo. Results show that Pb and Zn concentrations in soil samples from Mitrovica (1953-2576 mg kg −1) and Obiliqi regions (138-179 mg kg −1) and their bioaccumulation levels in onion were significantly higher in comparison with the control group. There was an adverse negative correlation between Pb or Zn concentration and ALA-D activity and total chlorophyll content, and a positive correlation with ALA content. This study indicates that ALA-D activity can be used as a very sensitive biomarker for evaluation of heavy metal pollution. The bioaccumulation of heavy metals from soil polluted areas poses a threat for food contamination and public health.

Exposure to Pb-halide perovskite nanoparticles can deliver bioavailable Pb but does not alter endogenous gut microbiota in zebrafish.

Lead-halide perovskite nanoparticles (NPs) are a new technology, and investigation of toxicity is of considerable importance due to the potential lead (Pb) release into the environment. The aim of the study was to investigate aqueous and dietary toxicity of Pb-halide perovskite NP and Pb in zebrafish Danio rerio. Perovskite NP toxicity was evaluated in zebrafish by mortality, gene expression, histopathology, and phylogenetic analysis of gut microbiota. Zebrafish larvae were exposed to five Pb-halide perovskite NPs in parallel with Pb(NO3)2 exposures, and zebrafish adults were exposed to the three perovskite NPs that caused the strongest effect and Pb(NO3)2. No median lethal concentration (LC50) was observed for zebrafish larvae exposed to up to 200mg/L of perovskite NPs for 96h. Mortality, metallothionein 2 (mt2) and δ-aminolevulinic acid dehydratase (ala-d) gene expression (24-h exposure) in zebrafish larvae after aqueous perovskite NPs exposures did not differ from total Pb concentration - response curves. The lack of differences in mortality and gene expression between perovskite NPs and soluble Pb after aqueous exposure suggest that toxicity from perovskite NPs can be attributed to bioavailable Pb rather than nano-specific effects. Induction of mt2 and reduction of ala-d expression levels in liver tissues showed Pb bioavailability after 2-d and 4-d dietary exposure to perovskite-spiked feeds. Changes in gut microbiota of adult zebrafish were detected after 14-d exposure to Pb-spiked food, but no changes were detected from perovskite-NP spiked food. The phylogenetic analysis identified different microbiome profiles of Pb-fed fish compared to perovskite-fed fish suggesting a different mechanism of toxicity. Exposure to Pb-halide perovskite NPs led to absorption of Pb likely from release of Pb ions rather than absorption of NPs. Pb-halide perovskite NPs can release bioavailable Pb and this needs to be considered during the development of this technology.

Effect of lead exposure and nutritional iron-deficiency on immune response: A vaccine challenge study in rats

The prevalence of iron (Fe) deficiency and subclinical lead (Pb) toxicity is high in developing countries like India, and information on their potential additive effects on immune responses is scant. The current study assessed immune parameters in dual Pb-exposed\\Fe-deficient weanling SD rats. Rats were fed a control (CD) or Fe-deficient (ID) diet for 4 weeks and then evaluated for hemoglobin (Hb) and serum Fe status. Then, half the rats in each group began to receive daily oral Pb exposure (25 mg/4 ml/kg BW; gavage) or vehicle for a further 4 weeks (while maintained on original respective diets). After the 4-weeks of dosing, rats were assessed for Hb and serum Fe, and for blood lead level (BLL) and δ-aminolevulinic acid dehydratase (ALAD) activity. At this point, half the rats in each group (now n = 8) were then vaccinated with tetanus toxoid (TT), and then two boosters at 2-week intervals. All the time, rats stayed on their original respective diets along with exposure to Pb on alternate days. At 2 weeks after the final booster, rats were euthanized and blood collected to assess total/specific IgG and IgM levels; mucosal (intestinal) IgA levels were also determined. Spleens were taken to assess CD4+ and CD8+ cell levels and for ex vivo measures of splenocyte proliferation/TH1 and TH2 cytokine formation. The results indicated significant lowering of Hb and serum Fe levels in ID rats and increased blood Pb and decreased ALAD activity in all Pb-exposed rats. Fe-deficiency alone induced significant increases in ALAD activity, but only in an absence of Pb. While there was no impact of any regimen on total or TT-specific IgG, significant decreases in mucosal IgA and TT-specific IgM were seen in ID-fed Pb-exposed rats. CD4+ cell levels were not impacted by treatment; CD8+ levels were increased in all ID/Pb-exposed rats. Ex-vivo splenocyte proliferation was significantly higher among vaccinated rats, as well as ID-fed Pb-exposed unvaccinated rats. Cytokine formation in all cases was highly variable. The results suggest that Fe deficiency compromised cell-mediated, mucosal, and/or humoral immune response-related endpoints and that Pb exposure during the deficiency further impacted these outcomes.