Kitazin-pea interaction: understanding the fungicide induced nodule alteration,cytotoxicity, oxidative damage and toxicity alleviation by Rhizobium leguminosarum

Realizing the severity of fungicidal toxicity to legumes and importance of fungicide tolerant rhizobia in legume production, kitazin tolerant (2400 μg mL⁻¹) strain RP1 was recovered from pea nodules and was identified as Rhizobium leguminosarum (accession no. {"type":"entrez-nucleotide","attrs":{"text":"KY940047","term_id":"1339307653","term_text":"KY940047"}}KY940047). R. leguminosarum produced indole acetic acid (80.5 ± 2.5 mL⁻¹), siderophores: salicylic acid (54 ± 7.3 μg mL⁻¹) and 2,3-dihydoxybenzoic acid (31.9 ± 2.7 μg mL⁻¹), α-ketobutyrate (51 ± 3.2 per mg per protein per hour), solubilized insoluble phosphate (29.5 ± 1.8 μg mL⁻¹) and secreted 29.5 + 2.6 μg mL⁻¹ exopolysaccharides, which, however, decreased consistently with gradually increasing kitazin concentrations. Beyond the tolerance level, kitazin caused structural damage and altered membrane integrity of RP1, as revealed under scanning (SEM) and confocal (CLSM) electron microscopy. Phytotoxicity of kitazin to peas was obvious under both in vitro and in vivo conditions. A significant reduction of 23, 68, 57 and 50% in germination, seedling vigor index, plumule length and radicle length was found at 2× kitazin compared to the control. Cellular damage and cytotoxicity induced by kitazin in membrane altered root cells was detected with acridine orange/propidium iodide (AO/PI) and Evans blue dye. A maximum increase of 1.72, 5.2, 9.3 and 1.72, 5.2, 9.3-fold in red and blue fluorescence was quantified at 1×, 2×, and 3× doses of kitazin, respectively. In contrast, application of R. leguminosarum RP1 alleviated toxicity and enhanced the length of plant organs, dry biomass, symbiotic attributes, photosynthetic pigments, nutrient uptake and grain features of peas comparatively uninoculated and fungicide-treated plants. Additionally, strain RP1 expressively reduced the antioxidant enzymes peroxidase, ascorbate peroxidase, guaiacol peroxidase, catalase and malondialdehyde contents by 10, 2.2, 11, 20 and 4% compared to stressed plants raised at 192 μg kg⁻¹ soil. Moreover, a decline of 19, 21 and 20% in proline content extracted from roots, shoots and grains, respectively was recorded for R. leguminosarum inoculated pea plants grown with 96 μg kg⁻¹ kitazin. Also, the SEM and CLSM of roots revealed the bacterial colonization. In conclusion, R. leguminosarum tolerated a higher level of kitazin, secreted plant growth promoting (PGP) bioactive molecules even under fungicide stress and significantly increased the performance of peas while reducing the levels of proline and antioxidant enzymes. So, it can safely be suggested to legume growers that RP1 strain could inexpensively be explored as an efficient biofertilizer for enhancing the production of legumes especially peas while growing even under fungicide (kitazin) enriched soils.

Realizing the severity of fungicidal toxicity to legumes and the importance of fungicide tolerant rhizobia in legume production, kitazin tolerant strain RP1 was recovered from pea nodules and was identified as Rhizobium leguminosarum.     

Preparation of a magnetic multiwalled carbon nanotube@polydopamine/zeolitic imidazolate framework-8 composite for magnetic solid-phase extraction of triazole fungicides from environmental water samples

A novel magnetic zinc-based zeolitic imidazolate framework (MMP/ZIF-8) has been prepared using a magnetic multiwalled carbon nanotube@polydopamine nanocomposite as the magnetic core and support. It was then used as an adsorbent for magnetic solid-phase extraction of triazole fungicides from environmental water samples. Successful synthesis of MMP/ZIF-8 was confirmed by material characterization, and the results showed that the synthetic composite has a high Brunauer–Emmett–Teller surface area (141.56 m² g⁻¹), large total pore volume (0.636 mL g⁻¹), and high superparamagnetism with a saturation magnetization of 44.1 emu g⁻¹. To evaluate the extraction performance of MMP/ZIF-8, the main parameters that affect the extraction efficiency were optimized. Under the optimal conditions, the developed method shows good linearity (R² ≥ 0.9915) in the concentration range 1–400 μg L⁻¹. Low limits of detection (0.08–0.27 μg L⁻¹, signal/noise = 3 : 1) and good precision (intraday relative standard deviation ≤ 7.73%, interday relative standard deviation ≤ 9.65%) are also achieved. The developed method was applied for analysis of triazole fungicides in environmental water samples.

A magnetic ZIF-8 composite was prepared based on Fe₃O₄/MWCNT@PDA and used as an adsorbent for MSPE of triazoles from environmental water samples.     

Screening of multiclass pesticide residues in maca and Moringa oleifera by a modified QuEChERS sample preparation procedure and UPLC-ESI-MS/MS analysis

In the present study, a modified QuEChERS (quick, easy, cheap, effective, rugged, and safe) method was proposed for the simultaneous analysis of 75 pesticides in maca and Moringa oleifera with ultra-performance liquid chromatography-electrospray ionization-tandem mass spectrometry (UPLC-ESI-MS/MS). The developed method was validated in accordance with linearity, linear range, limit of detection, limit of quantification, accuracy, precision, and matrix effect. Each analyte had good linearity (R² > 0.99) in the corresponding concentration range. The method LOD and LOQ values of all the analytes ranged from 0.01 μg kg⁻¹ to 303.35 μg kg⁻¹ and 0.03 μg kg⁻¹ to 1011.15 μg kg⁻¹, respectively. The recoveries (n = 6) of the analyzed pesticides were in the range of 75.92–113.43%. The RSDs of precision were between 0.60% and 7.36%. All matrix effect values ranged from 81.79% to 118.71% and 80.36% to 119.64% in maca and Moringa oleifera, respectively. The analysis of 103 samples showed the presence of isofenphos-methyl in some of them. The method had a good application prospect and could be used as a general approach for the quantitative determination of pesticide residues in food.

In the present study, a modified QuEChERS (quick, easy, cheap, effective, rugged, and safe) method was proposed for the simultaneous analysis of 75 pesticides in maca and Moringa oleifera with UPLC-ESI-MS/MS.     

Differential Effects of Insulin on Splanchnic and Peripheral Glucose Disposal after an Intravenous Glucose Load in Man

The present study was designed to investigate the mechanisms by which insulin regulates the disposal of an intravenous glucose load in man. A combined tracer-hepatic vein catheter technique was used to quantitate directly the components of net splanchnic glucose balance (NSGB), i.e., splanchnic glucose uptake and hepatic glucose output, and peripheral (extrasplanchnic) glucose uptake. Four different protocols were performed: (a) intravenous infusion of glucose alone (6.5 mg kg⁻¹ min⁻¹) for 90 min (control group); (b) glucose plus somatostatin (0.6 mg/h) and glucagon (0.8 ng kg⁻¹ min⁻¹; (c) glucose plus somatostatin, glucagon, and insulin (0.15 mU kg⁻¹ min⁻¹); and (d) glucose plus somatostatin, glucagon, and insulin (0.4 m U kg⁻¹ min⁻¹). In groups 2-4, arterial blood glucose was raised to comparable levels to those of controls (170 mg/dl) by a variable glucose infusion. In the control group, plasma insulin levels reached 40 μU/ml at 90 min. NSGB switched from a net output of 1.71±0.13 to a net uptake of 1.5-1.6 mg kg⁻¹ min⁻¹ due to a 90-95% suppression of hepatic glucose output (P < 0.01) and a 105-130% elevation of splanchnic glucose uptake (from 0.78±0.13 to 1.6-1.8 mg kg⁻¹ min⁻¹; P < 0.01). Peripheral glucose uptake rose by 150-160% (P < 0.01). In group 2, plasma insulin fell to <5 μU/ml. Net splanchnic glucose output initially rose twofold but later returned to basal values. This response was entirely accounted for by similar changes in hepatic glucose output since splanchnic glucose uptake remained totally unchanged in spite of hyperglycemia. In contrast, peripheral glucose uptake rose consistently by 100% (P < 0.01) despite insulin deficiency. In an additional group of experiments, glucose metabolism by the forearm muscle tissue was quantitated during identical conditions to those of group 2 (hyperglycemia plus insulin deficiency). Both the arterial-deep venous blood glucose difference and forearm glucose uptake increased markedly by 300-400% (P < 0.05 - <0.01). In group 3, plasma insulin was maintained at near-basal, peripheral levels (12-14 μU/ml). Hepatic glucose output decreased slightly by 35-40% (P < 0.05) while splanchnic glucose uptake remained unchanged. Consequently, the net glucose overproduction seen in group 2 was totally prevented although NSGB still remained as a net output. In group 4, peripheral insulin levels were similar to those of the control group (35-40 μU/ml). The suppression of hepatic glucose output was more pronounced (60-65%) and splanchnic glucose uptake rose consistently by 65% (P < 0.01). Consequently, NSGB did not remain as a net output but eventually switched to a small uptake (0.3 mg kg⁻¹ min⁻¹). Peripheral glucose uptake rose to the same extent as in controls.     

Efficient analyses of triazole fungicides in water,honey and soy milk samples by popping candy-generated CO2 and sugaring-out-assisted supramolecular solvent-based microextraction prior to HPLC determinations

An enrichment method, namely popping candy-generated CO₂ and sugaring-out-assisted supramolecular solvent-based microextraction (PGS-SUPRA), was investigated for the determination of triazole fungicide residues in water, honey and soy milk samples. The extraction process was carried out by adding popping candies into a centrifuge tube. Consequently, rapid dispersion and mass transfer of extractants can be achieved without using dispersants and auxiliary devices, and therefore, the extraction efficiency increased. The extraction parameters affecting the efficiency of the developed method were investigated. The presented method was then analysed by high-performance liquid chromatography. Under the selected condition, the wide linearity of triazole fungicides after preconcentration by the proposed microextraction method ranged from 30 to 1000 μg L⁻¹ for triadimefon and from 90 to 1000 μg L⁻¹ for myclobutanil, tebuconazole and hexaconazole, with a coefficient for determination (R²) greater than 0.992. The limits of detection (LODs) and limits of quantitation (LOQs) were in the range of 10–30 μg L⁻¹ and 30–90 μg L⁻¹, respectively. The precisions were assessed from the relative standard deviations (RSDs) of the retention time and peak area obtained from intra- (n = 3) and inter-day (n = 3 × 5) experiments, and were greater than 1.66% and 13.52%, respectively. Moreover, the proposed method provided high enhancement factors (EnFs) ranging from 14 to 51 folds. This technique has been prosperously applied for the extraction of fungicide residues in water, honey and soy milk samples with a recovery within the range of 60–114%. Overall, the developed method was found to be advantageous as compared with other sample preparation methods.

Schematic diagram of the popping candy-generated CO₂ and sugaring-out-assisted supramolecular solvent-based microextraction (PGS-SUPRA) method prior to HPLC analysis.     

Unraveling the potential of pesticide-tolerant Pseudomonas sp. augmenting biological and physiological attributes of Vigna radiata (L.) under pesticide stress

In the agricultural domain, chemical pesticides are repetitively and harshly used to kill harmful pests, but they often pose a serious threat to microbial diversity, soil fertility and agricultural output. To deal with these problems, pesticide-tolerant plant growth promoting (PGP) rhizobacterial strains are often used to combat pesticidal toxicity. Here, Pseudomonas sp. PGR-11 (accession no. {"type":"entrez-nucleotide","attrs":{"text":"OM348534","term_id":"2182243475","term_text":"OM348534"}}OM348534), recovered from a Vigna radiata (L.) rhizosphere, produced various growth regulating (GR) substances, including indole-3-acetic acid (IAA; 82.5 ± 9.2 μg mL⁻¹), enzyme 1-aminocyclopropane 1-carboxylate (ACC) deaminase (μM α-ketobutyrate mg⁻¹ protein h⁻¹), siderophores and ammonia. Strain PGR-11 grew well when cultured in growth medium with added metalaxyl (MTXL; 1200 μg mL⁻¹), carbendazim (CBZM; 800 μg mL⁻¹) and tebuconazole (TBZL; 1600 μg mL⁻¹). Pseudomonas sp. synthesized PGP substances even in the presence of increasing doses of pesticides. The phytotoxicity of the tested pesticides was assessed both in vitro and under pot-house conditions using a Vigna radiata (L.) crop. Increasing concentrations of chemical pesticides negatively impacted the growth, physiological and biochemical features. However, pesticide-tolerant Pseudomonas sp. relieved the toxicity and improved the biological attributes of the plant. Bio-inoculated plants showed significant enhancement in germination attributes, dry biomass, symbiotic features and yield features when compared to un-inoculated ones. Furthermore, with 100 μg metalaxyl kg⁻¹ soil, strain PGR-11 increased the chl-a, chl-b, total chlorophyll, carotenoids, SPAD index, photosystem efficiency (Fv/Fm), PSII quantum yield (FPSII), photochemical quenching (qP) and non-photochemical quenching (NpQ) content by 12, 19, 16, 27, 34, 41, 26, 29 and 33%, respectively, over un-inoculated but pesticide-treated plants. Additionally, inoculation of Pseudomonas sp. with 100 μg tebuconazole kg⁻¹ soil caused a significant (p ≤ 0.05) enhancement in transpiration rate (E), stomatal conductance (g_s), photosynthetic rate (P_N), vapor pressure deficit (kPa) and internal CO₂ concentration (C_i) of 19, 26, 23, 28 and 34%, respectively. Conclusively, the power to tolerate abnormally high pesticide concentration, the capacity to produce/secrete PGP substances even in a pesticide-stressed medium and the potential for improving/increasing the growth and physiology of plants by pesticide detoxification makes Pseudomonas sp. PGR-11 a fascinating choice for augmenting the productivity of V. radiata (L.) even in pesticide-stressed soils. The current findings will be helpful for exploring pesticide-tolerant ACC-deaminase-positive microbial strains as gifted entities for the environmental bioremediation of pesticides.

In the agricultural domain, chemical pesticides are repetitively and harshly used to kill harmful pests, but they often pose a serious threat to microbial diversity, soil fertility and agricultural output.     

Arbuscular mycorrhizal inoculation increases molybdenum accumulation but decreases molybdenum toxicity in maize plants grown in polluted soil

Molybdenum (Mo) is an important micronutrient required by both plants and microorganisms, but may become toxic when presents in excess concentration. However, Mo toxicity in soil-plant systems as influenced by arbuscular mycorrhizal (AM) fungi (AMF) still remains unknown. Here, a pot culture experiment was conducted to study the effects of inoculation with Claroideoglomus etunicatum BEG 168 on the growth and Mo content of maize plants growing in soil supplemented with different levels (0, 1000, 2000, and 4000 mg kg⁻¹) of Mo. Results show that the added Mo had no significant effects on AM colonization rate, which ranged from 77% to 92%. Mo addition decreased plant dry weights and leaf pigment contents, as well as nutrient uptake of P, N, Fe, Mg and Cu in shoots and roots, and in most cases, the highest level (4000 mg kg⁻¹) showed the most inhibitory effects. Overall, AM inoculation enhanced plant growth, mineral nutrient uptake, leaf pigment contents and photosynthetic rate under all Mo addition levels. Mo concentrations in plants without Mo addition ranged from 13.1 to 40.1 mg kg⁻¹ in roots, and from 42.8 to 58.4 mg kg⁻¹ in shoots. Addition of Mo increased Mo concentrations in both shoots and roots of all the plants, but showed no significant dose-dependent effects. In non-inoculated plants receiving Mo addition, Mo concentrations were not lower than 400 mg kg⁻¹ in shoots and higher than 1300 mg kg⁻¹ in roots respectively. AM inoculation further enhanced Mo concentrations in shoots and roots, but decreased shoot/root Mo ratio at 2000 and 4000 mg kg⁻¹ Mo addition levels. In AM inoculation treatments, soil pH exhibited a decreasing trend with increasing Mo addition level. In conclusion, excess Mo caused toxicity in maize plants, while AM fungus C. etunicatum BEG 168 was tolerant to the added Mo, and could alleviate the Mo-induced phytotoxicity by improving plants'' mineral nutrition, leaf pigment contents and photosynthetic properties, and by mediating Mo partitioning in plants and soil pH. Our present results suggest a specific protection mechanism exists in AM plants against excess Mo, and their promising potential in ecological restoration and phytoremediation of Mo-polluted sites.

Arbuscular mycorrhizal inoculation increases Mo accumulation but decreases Mo phytotoxicity in maize plants grown in polluted soil.     

Influence of Ni content on structural,magnetocaloric and electrical properties in manganite La0.6Ba0.2Sr0.2Mn1−xNixO3 (0 ≤ x ≤ 0.1) type perovskites

We present a detailed study on the physical properties of La_0.6Ba_0.2Sr_0.2Mn_1−xNi_xO₃ samples (x = 0.00, 0.05 and 0.1). The ceramics were fabricated using the sol–gel route. Structural refinement, employing the Rietveld method, disclosed a rhombohedral R$\bar{3}$c phase. The magnetization vs. temperature plots show a paramagnetic–ferromagnetic (PM–FM) transition phase at the T_C (Curie temperature), which decreases from 354 K to 301 K. From the Arrott diagrams M²vs. μ₀H/M, we can conclude the phase transition is of the second order. Based on measurements of the isothermal magnetization around T_C, the magnetocaloric effects (MCEs) have been calculated. The entropy maximum change (−ΔS_M) values are 7.40 J kg⁻¹ K⁻¹, 5.6 J kg⁻¹ K⁻¹ and 4.48 J kg⁻¹ K⁻¹, whereas the relative cooling power (RCP) values are 232 J kg⁻¹, 230 J kg⁻¹ and 156 J kg⁻¹ for x = 0.00, 0.05 and 0.10, respectively, under an external field (μ₀H) of 5 T. Through these results, the La_0.6Ba_0.2Sr_0.2Mn_1−xNi_xO₃ (0 ≤ x ≤ 0.1) samples can be suggested for use in magnetic refrigeration technology above room temperature. The electrical resistivity (ρ) vs. temperature plots exhibit a transition from metallic behavior to semiconductor behavior in the vicinity of T_M–SC. The adiabatic small polaron hopping (ASPH) model is applied in the PM-semiconducting part (T > T_MS). Throughout the temperature range, ρ is adjusted by the percolation model. This model is based on the phase segregation of FM-metal clusters and PM-insulating regions.

We present a detailed study on the physical properties of La_0.6Ba_0.2Sr_0.2Mn_1−xNi_xO₃ samples (x = 0.00, 0.05 and 0.1).     

Micranthanosides I and II,two novel 1,10-secograyanane diterpenoids and their antinociceptive analogues from the leaves and twigs of Rhododendron micranthum

Micranthanosides I and II (1–2), two diterpenoid glucosides featuring a new 1,10-secograyanane skeleton, thirteen new diterpenoid glycosides (3–15), and 21 known analogues were obtained from the ethanol extract of the leaves and twigs of Rhododendron micranthum. Micranthanoside XII (12) represent the first example of 3,5-epoxy-4,5-seco-ent-kaurane diterpenoid. The structures of these compounds were determined by spectroscopic data analysis and quantum chemical calculations. To clarify the chemical basis and provide reference for rational use of this medicinal plant, the antinociceptive and the anti-inflammatory activities of the compounds were tested. In the acetic acid-induced writhing test, compounds 17 and 19 showed significant antinociceptive activity at a dose of 3 mg kg⁻¹ and compounds 2, 6 and 32 showed significant antinociceptive activity at a dose of 10 mg kg⁻¹. Toxic reactions such as nausea and convulsion were observed when 17, 19, 29, and 31 at a dose of 10 mg kg⁻¹ or 30 and 33 at a dose of 1 mg kg⁻¹ were administered. The anti-inflammatory activities of the isolated compounds were evaluated by measuring the inhibitory effects of LPS-induced NO production in BV2 cells. At 10 μM, micranthanoside IX (9) and rhodomicranoside F (26) showed moderate anti-inflammatory activities with inhibition rates of 56.31% and 72.43%, respectively.

Two unusual diterpenoids with a new 1,10-secograyanane skeleton and thirteen new analogues from the leaves and twigs of Rhododendron micranthum were obtained.     

Novel chalcone derivatives containing a 1,2,4-triazine moiety: design,synthesis, antibacterial and antiviral activities

A series of novel chalcone derivatives containing the 1,2,4-triazine moiety were synthesized and their structures were confirmed by ¹H NMR, ¹³C NMR and elemental analyses. Antiviral bioassays revealed that most of the compounds exhibited good antiviral activity against tobacco mosaic virus (TMV) at a concentration of 500 μg mL⁻¹. The designated compound 4l was 50% effective in terms of curative and protective activities against TMV with 50% effective concentrations (EC₅₀) of 10.9 and 79.4 μg mL⁻¹, which were better than those of ningnanmycin (81.4 and 82.2 μg mL⁻¹). Microscale thermophoresis (MST) also showed that the binding of compound 4l to coat protein (TMV-CP) yielded a K_d value of 0.275 ± 0.160 μmol L⁻¹, which was better than that of ningnanmycin (0.523 ± 0.250 μmol L⁻¹). At the same time, molecular docking studies for 4l with TMV-CP (PDB code:1EI7) showed that the compound was embedded well in the pocket between the two subunits of TMV-CP. Meanwhile, compound 4a demonstrated excellent antibacterial activities against Ralstonia solanacearum (R. solanacearum), with an EC₅₀ value of 0.1 μg mL⁻¹, which was better than that of thiodiazole-copper (36.1 μg mL⁻¹) and bismerthiazol (49.5 μg mL⁻¹). The compounds act by causing folding and deformation of the bacterial cell membrane as observed using scanning electron microscopy (SEM). The chalcone derivatives thus synthesized could become potential alternative templates for novel antiviral and antibacterial agents.

A series of novel chalcone derivatives containing the 1,2,4-triazine moiety were synthesized and their structures were confirmed by ¹H NMR, ¹³C NMR and elemental analyses.     

Electrochemical oxidation of resorcinol: mechanistic insights from experimental and computational studies

This work investigates the mechanisms of resorcinol oxidation by density functional theory (DFT) calculation and cyclic voltammetry measurements. Complementary data from experimental and computational studies provide new insights into the reaction mechanisms. At both macro- and micro-electrodes, cyclic voltammetry of resorcinol is chemically and electrochemically irreversible over the whole pH range (1–14). Resorcinol molecules undergo a 1H⁺ 1e⁻ oxidation at pH < pK_a1 and a 1e⁻ oxidation at pH > pK_a2 to form radicals. The radicals then readily react to form dimers/polymers deposited on the electrode surface. All of the experimental findings are consistent with the proposed mechanisms, including the apparent transfer coefficient (β) of 0.6 ± 0.1, the slope of the peak potential (E_p) against pH of −54 mV pH⁻¹, the peak-shaped responses at micro-electrodes, and the fouling of the electrodes upon the oxidation of resorcinol. DFT calculation of the reaction energy of elementary steps and the eigenvalues of the highest occupied molecular orbital (HOMO) of the radical intermediates confirms that the (1H⁺) 1e⁻ oxidation is the energetically favorable pathway. In addition to mechanistic insights, an electrochemical sensor is developed for resorcinol detection at microelectrodes in low ionic strength samples with the sensitivity of 123 ± 4 nA μM⁻¹ and the limit of detection (3 s_B m⁻¹) of 0.03 μM.

Resorcinol oxidation mechanism was investigated by DFT calculation and cyclic voltammetry experiments at macro- and micro-electrodes (1 ≤ pH ≤ 14).     

Heavy metal mediated phytotoxic impact on winter wheat: oxidative stress and microbial management of toxicity by Bacillus subtilis BM2

Heavy metals are toxic environmental contaminants, which severely affect microbial composition and functions and, concurrently, crop production. Due to these issues, the present study focussed on the selection of metal tolerant microbes endowed with metal detoxification abilities and their role in the management and remediation of metal contaminated soils. The metal tolerant bacterium BM2, identified as Bacillus subtilis by 16SrRNA gene sequencing, survived well under metal pressure and tolerated 1600 and 2000 μg mL⁻¹ of Ni and Pb, respectively. The inhibitory impact of metals on wheat increased consistently with a progressive increase in metal concentration. Deposition of Ni and Pb within root and leaf and oxidative stress were validated by SEM, EDX and CLSM. The overall growth parameters of wheat grown under metal stress were improved following B. subtilis BM2 colonization. As an example, B. subtilis with 195 mg Pb kg⁻¹ enhanced the length and dry biomass of shoots by 14% and 23%, respectively, over the control. Also, strain BM2 improved the grain yield significantly by 49% at 870 mg Ni kg⁻¹ and by 50% at 585 mg Pb kg⁻¹ compared to uninoculated plants. Moreover, B. subtilis BM2 relieved the metal stress on wheat and caused a significant drop in proline and malondialdehyde content and the activities of antioxidant enzymes, like catalase (CAT), superoxide dismutase (SOD) and glutathione reductase (GR). This study, therefore, provided solutions to the metal toxicity problems faced by winter wheat and clearly suggests that the metal detoxification potential of B. subtilis BM2 could be greatly useful in the management of metal polluted soils.

The bacterial management of heavy metal toxicity to durum wheat under a metal stressed environment was demonstrated.     

Copper-doped lanthanum manganite La0.65Ce0.05Sr0.3Mn1−xCuxO3 influence on structural,magnetic and magnetocaloric effects

Bulk nanocrystalline samples of La_0.65Ce_0.05Sr_0.3Mn_1−xCu_xO₃ (0 ≤ x ≤ 0.15) manganites are prepared by the sol–gel based Pechini method. The effect of the substitution for Mn with Cu upon the structural and magnetic properties has been investigated by means of X-ray diffraction (XRD), Raman spectroscopy and dc magnetization measurements. The structural parameters obtained using Rietveld refinement of XRD data showed perovskite structures with rhombohedral (R$\bar{3}$c) symmetry without any detectable impurity phase. Raman spectra at room temperature reveal a gradual change in phonon modes with increasing copper concentration. The analysis of the crystallographic data suggested a strong correlation between structure and magnetism, for instance a relationship between a distortion of the MnO₆ octahedron and the reduction in the Curie temperature, T_c. A paramagnetic to ferromagnetic phase transition at T_C is observed. The experimental results confirm that Mn-site substitution with Cu destroys the Mn³⁺–O²⁻–Mn⁴⁺ bridges and weakens the double exchange (DE) interaction between Mn³⁺ and Mn⁴⁺ ions, which shows an obvious suppression of the FM interaction in the La_0.65Ce_0.05Sr_0.3Mn_1−xCu_xO₃ matrix. The maximum magnetic entropy change −ΔS^max_M is found to decrease with increasing Cu content from 4.43 J kg⁻¹ K⁻¹ for x = 0 to 3.03 J kg⁻¹ K⁻¹ for x = 0.15 upon a 5 T applied field change.

Bulk nanocrystalline samples of La_0.65Ce_0.05Sr_0.3Mn_1−xCu_xO₃ (0 ≤ x ≤ 0.15) manganites are prepared by the sol–gel based Pechini method.     

Impact of synthesis route on structural,magnetic, magnetocaloric and critical behavior of Nd0.6Sr0.4MnO3 manganite

Nd_0.6Sr_0.4MnO₃ polycrystalline manganite was synthesized by two different methods: the auto-combustion reaction (NSMO-AC) and the sol–gel method (NSMO-SG). The structural, magnetic, magnetocaloric and critical behavior of the samples were examined. Rietveld refinements of the XRD patterns revealed that both compounds are pure single phase indexed to the orthorhombic system adopting the Pnma space group. The nanometric size estimated using the Williamson–Hall method was confirmed by TEM micrographs. Magnetic measurements as a function of temperature indicated that both samples underwent a second order ferromagnetic (FM)–paramagnetic (PM) phase transition at Curie temperature (T_C). The relative cooling power was observed to be around 95.271 J kg⁻¹ for NSMO-AC and 202.054 J kg⁻¹ for NSMO-SG at μ₀H = 5 T, indicating that these materials are potential candidates for magnetic refrigeration application close to room temperature. The critical behavior was estimated using diverse techniques based on the isothermal magnetization data recorded around the critical temperature T_C. The calculated values are fully satisfactory to the requirements of the scaling theory, implying their reliability. The estimated critical exponents matched well with the values anticipated for the mean-field model and the 3D Ising model for NSMO-AC and NSMO-SG, respectively, showing that the magnetic interactions depended on the process of elaboration.

The relative cooling power is around 95.271 J kg⁻¹ for NSMO-AC and 202.054 J kg⁻¹ for NSMO-SG at μ₀H = 5 T, making these materials potential candidates for magnetic refrigeration application near room temperature.     

Branched-chain Amino Acid Nitrogen Transfer to Alanine In Vivo in Dogs: DIRECT ISOTOPIC DETERMINATION WITH [15N]LEUCINE

To investigate the contribution of branched-chain amino acids as a nitrogen source for alanine in vivo, dogs were infused with l-[¹⁵N]leucine, l-[U-¹⁴C]leucine, l-[2,3,3,3-²H₄]alanine, and d-[6,6-²H₂]-glucose. ¹⁴C and ¹⁵N isotopic equilibrium in plasma leucine, and deuterium enrichment in arterial and femoral plasma glucose and alanine were achieved within 3 h of initiation of the respective isotope infusion in all animals. The average flux of leucine determined by [¹⁵N]leucine was 5.4 μmol·kg⁻¹·min⁻¹, whereas using [¹⁴C]leucine it was 3.7 μmol·kg⁻¹·min⁻¹. Turnover rates for alanine and glucose were 11.0 and 17.2 μmol·kg⁻¹·min⁻¹, respectively.     

Enhanced mechanical properties of hBN–ZrO2 composites and their biological activities on Drosophila melanogaster: synthesis and characterization

In this study, six compositions in the system [x(h-BN)–(100 − x)ZrO₂] (10 ≤ x ≤ 90) were synthesized by a bottom up approach, i.e., the solid-state reaction technique. XRD results showed the formation of a novel and main phase of zirconium oxynitrate ZrO(NO₃)₂ and SEM exhibited mixed morphology of layered and stacked h-BN nanosheets with ZrO₂ grains. The composite sample 10 wt% h-BN + 90 wt% ZrO₂ (10B90Z) showed outstanding mechanical properties for different parameters, i.e., density (3.12 g cm⁻³), Young''s modulus (10.10 GPa), toughness (2.56 MJ m⁻³), and maximum mechanical strength (227.33 MPa). The current study further checked the in vivo toxicity of composite 10B90Z and composite 90B10Z using Drosophila melanogaster. The composite 10B90Z showed less cytotoxicity in this model, while the composite 90B10Z showed higher toxicity in terms of organ development as well as internal damage of the gut mostly at the lower concentrations of 1, 10, and 25 μg mL⁻¹. Altogether, the current study proposes the composite 10B90Z as an ideal compound for applications in biomedical research. This composite 10B90Z displays remarkable mechanical and biological performances, due to which we recommend this composition for various biomedical applications.

In this study, six compositions in the system [x(h-BN)–(100 − x)ZrO₂], (10 ≤ x ≤ 90) were synthesized by a bottom up approach, i.e., the solid-state reaction technique.     

Analysis method development and health risk assessment of pesticide and heavy metal residues in Dendrobium Candidum

Herbal medicines that are widely used worldwide are easily contaminated by pesticides and heavy metals, threatening human health. In this study, a modified QuEChERS pre-treatment method combined with HPLC/GC-MS/MS was established for the determination of 24 pesticide residues in Dendrobium candidum. The average recoveries of 24 pesticides in D. candidum were 76.9–110.0% with the relative standard deviation (RSD) of 0.28–11.40%, and their limits of detection (LOD) and limits of quantitation (LOQ) were 0.005–10 and 0.011–22 μg kg⁻¹, respectively. The results showed that 83.33% of all samples had detected pesticide residues with the concentrations of 0.06–312.83 μg kg⁻¹. Meanwhile, microwave digestion combined with ICP-MS was used to detect the residues of 8 heavy metals in D. candidum. The average recoveries of 8 heavy metals were 82.7–108.1% with an RSD of 1.4–8.0%, and their LOD and LOQ were 0.0001–0.05 mg kg⁻¹ and 0.0003–0.2 mg kg⁻¹, respectively. The results indicated that 8 heavy metals were all detected in all samples, and the highest concentration of Zn was 11.97 mg kg⁻¹. Furthermore, the health risk assessment showed that the risk of the detected pesticides and heavy metals in samples to humans, specifically to the general population including adults and children, was acceptable.

Residues of 24 pesticides and 8 heavy metals in Dendrobium candidum were determinated using both the modified QuEChERS-HPLC(GC)-MS/MS and microwave digestion-ICP-MS analysis methods, and their health risk was acceptable.     

Pharmacological Properties of NAI-603, a Well-Tolerated Semisynthetic Derivative of Ramoplanin

NAI-603 is a ramoplanin derivative designed to overcome the tolerability issues of the parent drug as a systemic agent. NAI-603 is highly active against aerobic and anaerobic Gram-positive bacteria, with MICs ranging from 0.008 to 8 μg/ml. MICs were not significantly affected by pH (range, 6 to 8), by inoculum up to 10⁸ CFU/ml, or by addition of 50% human serum. Against staphylococci and enterococci, NAI-603 was rapidly bactericidal, with minimum bactericidal concentration (MBC)/MIC ratios never exceeding 4. The frequency of spontaneous resistance was low at 2× to 4× MIC (≤1 × 10⁻⁶ to ≤1 × 10⁻⁸) and below the detection limit (about ≤1 × 10⁻⁹) at 8× MIC. Serial subcultures at 0.5× MIC yielded at most an 8-fold increase in MICs. In a systemic infection induced by methicillin-resistant Staphylococcus aureus (MRSA), the 50% effective dose (ED₅₀) of intravenous (i.v.) NAI-603 was 0.4 mg/kg, lower than that of oral (p.o.) linezolid (1.4 mg/kg) and subcutaneous (s.c.) teicoplanin (1.4 mg/kg) or vancomycin (0.6 mg/kg). In neutropenic mice infected with vancomycin-resistant enterococci (VRE), the ED₅₀s for NAI-603 were 1.1 to 1.6 mg/kg i.v., compared to 0.5 mg/kg i.v. of ramoplanin. The bactericidal activity was confirmed in vivo in the rat granuloma pouch model induced by MRSA, where NAI-603, at 40 mg/kg i.v., induced about a 2- to 3-log₁₀-reduction in viable bacteria in the exudates, which persisted for more than 72 h. The pharmacokinetic (PK) profiles of NAI-603 and ramoplanin at 20 mg/kg show similar half-lives (3.27 and 3.80 h, respectively) with the maximum concentration (C_max) markedly higher for NAI-603 (207 μg/ml versus 79 μg/ml). The favorable pharmacological profile of NAI-603, coupled with the absence of local tolerability issues, supports further investigation.     

Mezlocillin Pharmacokinetics After Single Intravenous Doses to Patients with Varying Degrees of Renal Function

The pharmacokinetics of mezlocillin were examined after single 2- and 4-g intravenous injections to three groups of male patients with creatinine clearances of I ≥ 60, II = 21 to 59, and III ≤ 20 ml min⁻¹ 1.73 m⁻². The decline in serum antibiotic levels was biphasic in all groups, and serum data were interpreted in terms of the pharmacokinetic two-compartment model. The mean elimination half-life of mezlocillin after the 2-g dose was 1.3, 1.5, and 2.3 h in groups I, II, and III, respectively. Equivalent values after the 4-g dose were 1.2, 1.6, and 4.4 h. In three functionally anephric patients the mean serum half-life of mezlocillin was 1.5 h during hemodialysis. Mean antibiotic levels in serum were greater than 10 μg ml⁻¹ for 4 h after the 2- and 4-g doses in group I and 8 h in group II. In group III, levels greater than 10 μg ml⁻¹ were maintained for 6 h after the 2-g dose and over 12 h after the 4-g dose. Mezlocillin distribution characteristics were largely independent of renal function and dose size. The only observable change occurred in the value of V_dss, which was significantly increased to 0.32 with 0.38 liter kg⁻¹ in severe renal impairment, compared to ca. 0.2 liter kg⁻¹ in subjects with normal or slightly impaired renal function. Cumulative 24-h urinary excretion accounted for 50, 40, and 3.2% of the dose in groups I, II, and III, respectively. Urine levels of mezlocillin were uniformly greater than the minimum inhibitory concentration for susceptible organisms for 12 h after dosing in all patients who produced urine. Because of the relatively small increase in the mezlocillin elimination half-life with declining renal function, dose reduction is necessary only in cases of severe renal impairment.     

An investigation of the metabolic activity,isozyme contribution,species differences and potential drug–drug interactions of PI-103, and the identification of efflux transporters for PI-103-O-glucuronide in HeLa1A9 cells

PI-103 is a phosphatidylinositol 3-kinase inhibitor that includes multiple receptor affinity modifications, and it is also a therapeutic drug candidate primarily for human malignant tumors. However, its metabolic fate and potential drug–drug interactions involving human cytochrome P450 (CYP) and UDP-glucuronosyltransferases (UGT) enzymes remain unknown. In this study, our results demonstrated that the intrinsic clearance (CL_int) values of oxidated metabolite (M1) in human liver microsomes (HLM) and human intestine microsomes (HIM) were 3.10 and 0.08 μL min⁻¹ mg⁻¹, respectively, while PI-103 underwent efficient glucuronidation with CL_int values of 15.59 and 211.04 μL min⁻¹ mg⁻¹ for mono-glucuronide (M2) by HLM and HIM, respectively. Additionally, reaction phenotyping results indicated that CYP1A1 (51.50 μL min⁻¹ mg⁻¹), 1A2 (46.96 μL min⁻¹ mg⁻¹), and UGT1A1 (18.80 μL min⁻¹ mg⁻¹), 1A7 (8.52 μL min⁻¹ mg⁻¹), 1A8 (8.38 μL min⁻¹ mg⁻¹), 1A9 (34.62 μL min⁻¹ mg⁻¹), 1A10 (107.01 μL min⁻¹ mg⁻¹) were the most important contributors for the oxidation and glucuronidation of PI-103. Chemical inhibition assays also suggest that CYP1A2 and UGT1A1, 1A9 play a predominant role in the metabolism of PI-103 in HLM. Significant activity correlations were detected between phenacetin-N-deacetylation and M1 (r = 0.760, p = 0.004) as well as β-estradiol-3-O-glucuronide and M2 (r = 0.589, p = 0.044), and propofol-O-glucuronidation and M2 (r = 0.717, p = 0.009). Furthermore, the metabolism of PI-103 revealed marked species differences, and dogs, rats, mice and mini-pigs were not the appropriate animal models. Gene silencing of breast cancer resistance protein (BCRP) or multidrug resistance-associated protein (MRPs) transporter results indicated that M2 was mainly excreted by BCRP, MRP1 and MRP4 transporters. Moreover, PI-103 displayed broad-spectrum inhibition towards human CYPs and UGTs isozymes with IC₅₀ values ranging from 0.33 to 6.89 μM. Among them, PI-103 showed potent non-competitive inhibitory effects against CYP1A2, 2C19, 2E1 with IC₅₀ and K_i values of less than 1 μM. In addition, PI-103 exhibited moderate non-competitive inhibition against UGT1A7, 2B7, and moderate mixed-type inhibition towards CYP2B6, 2C9 and UGT1A3. Their IC₅₀ and K_i values were 1.16–6.89 and 0.56–5.64 μM, respectively. In contrast, PI-103 could activate the activity of UGT1A4 in a mechanistic two-site model with a K_i value of 13.76 μM. Taken together, PI-103 was subjected to significant hepatic and intestinal metabolism. CYP1A1, 1A2 and UGT1A1, 1A7, 1A8, 1A9, 1A10 were the main contributing isozymes, whereas BCRP, MRP1 and MRP4 contributed most to the efflux excretion of M2. Meanwhile, PI-103 had a potent and broad-spectrum inhibitory effect against human CYPs and UGTs isozymes. These findings could improve understanding of the metabolic fates and efflux transport of PI-103. The inhibited human CYP and UGT activities could trigger harmful DDIs when PI-103 is co-administered with clinical drugs primarily cleared by these CYPs or UGTs isoforms. Additional in vivo studies are required to evaluate the clinical significance of the data presented herein.

Metabolic activity and disposition characteristics of PI-103.