Nickel uptake in leafy greens from contaminated soil: an investigation into phytoavailability and health risk assessment using in vitro digestion model.

Nickel (Ni) is a toxic metal that not only pollutes the environment but also causes harmful impacts on plant growth and human health. Therefore, it is crucial to assess the relationship between the phytoavailability of Ni in soil and its accumulation in edible and non-edible parts of vegetables. A pot experiment was conducted to investigate Ni uptake in three different leafy vegetables, spinach (Spinacia oleracea L.), lettuce (Lactuca sativa L.), and fenugreek (Trigonella foenum-graecum L.), grown in soil artificially contaminated with Ni at three different treatment levels (100 mg kg-1, 200 mg kg-1, and 300 mg kg-1). The potential dietary toxicity of these vegetables in humans was examined by using an in vitro digestion model. The lowest and highest chlorophyll contents were detected in lettuce at 300 mg kg-1 of Ni concentration and in control plants of spinach. Their values were 34.16 ± 3.01 (SPAD unit) and 53 ± 3.7673 (SPAD unit), respectively. Among the three vegetables, lettuce and spinach at 300 mg kg-1 exhibited the highest accumulation of Ni, with 43 mg kg-1 in edible parts and 182 mg kg-1 in non-edible parts. Furthermore, health risk index (HRI) values were found to be > 1 for lettuce and fenugreek at Ni concentrations of 200 and 300 mg kg-1 for both children and adults. The average bioaccessibility of Ni in lettuce, fenugreek, and spinach during the gastrointestinal phase was 32-23%, 24-10%, and 45-37%, respectively, at a Ni concentration of 300 mg kg-1. All three vegetables grown on Ni-contaminated soil may potentially contribute to food chain toxicity. The HRI values being > 1 suggest that these vegetables are unsafe for consumption. Monitoring of Ni concentration in leafy vegetables is essential to minimize human health risks associated with food chain contamination.

Characterization of functional genes GS3 and GW2 and their effect on the grain size of various landraces of rice (Oryza sativa).

BACKGROUND: Grain size is an essential factor of grain quality and yield in rice. The genetic studies have substantially contributed to enhancing yield and maintaining a good quality of rice. The two major genes GS3 (a negative regulator of grain length) and GW2 (a negative regulator of grain width) with functional mutation play a significant role in controlling the grain size of rice. METHODS AND RESULTS: In the study, 17 different widely grown Pakistani landraces of various genetic and geographic backgrounds were evaluated for grain phenotypic traits (1000-grain weight, length, width, and thickness) and also screened for genotypic mutation in GS3 and GW2 genes. Phenotypic data revealed the range for grain weight from 16.86 g (Lateefy) to 26.91 g (PS2), grain length ranged from 7.27 mm (JP-5) to 12.18 mm (PS2), grain width ranged from 2.01 mm (Lateefy) to 3.51 mm (JP5), and grain thickness ranged from 1.79 mm to 2.19. Correlation revealed a negative and significant correlation between grain width and length. There was no significant correlation between grain length and 1000-grain weight and grain width. LSD test displayed that the means of three variables grain length, grain width, and 1000-grain weight were statistically different from one another except grain width and grain breadth. Fifteen accessions carried the domesticated allele of GS3 while JP5 and Fakhr-e-Malakand carried the dominant allele. Similarly, fifteen accessions carried the dominant allele of GW2 while JP-5 and Fakhr-e-Malakand carried the mutant allele. CONCLUSIONS: The study shows that the mutant alleles of both genes are of significance to pyramid them in any breeding program. However, just incorporating favorable alleles is not the sole solution for improving the grain size. Therefore, further elucidation of GS3 and GW2 genes regulatory network, their interaction, trade-off, and pathways will better coordinate their marker-assisted selection in the future breeding program. Additionally, the study concluded that the selection of grain size was not dependent on 1000-grain weight in the selected germplasm.

Evaluation of Jatropha curcas L. leaves mulching on wheat growth and biochemical attributes under water stress.

BACKGROUND: Organic mulches are widely used in crop production systems. Due to their benefits in improving soil fertility, retention of soil moisture and weed control. Field experiments were conducted during wheat growing seasons of 2018-2019 and 2019-2020 to evaluate the effects of Jatropha leaves mulch on the growth of wheat varieties 'Wadan-17' (rainfed) and 'Pirsabaq-2013' (irrigated) under well irrigated and water stress conditions (non-irrigated maintaining 40% soil field capacity). Jatropha mulch was applied to the soil surface at 0, 1, 3 and 5 Mg ha-1 before sowing grains in the field. Under conditions of water stress, Jatropha mulch significantly maintained the soil moisture content necessary for normal plant growth. RESULTS: We noted a decrease in plant height, shoot and root fresh/dry weight, leaf area, leaf relative water content (LRWC), chlorophyll, and carotenoid content due to water stress. However, water stress caused an increase in leaf and root phenolics content, leaf soluble sugars and electrolytes leakage. We observed that Jatropha mulch maintained LRWC, plant height, shoot and root fresh/dry weight, leaf area and chlorophyll content under water stress. Moreover, water stress adverse effects on leaf soluble sugar content and electrolyte leakage were reversed to normal by Jatropha mulch. CONCLUSION: Therefore, it may be concluded that Jatropha leaves mulch will minimize water stress adverse effects on wheat by maintaining soil moisture and plant water status.

Toxic effect of common poisonous plants of district Bannu, Khyber Pakhtunkhwa, Pakistan.

The present paper was a part of Ph.D research work, conducted during the year 2014, in which 87 poisonous plants belonging to 54 genera, were collected, documented and preserved in the herbarium of Bannu, Department of Botany UST, Bannu Khyber Pakhtunkwa Pakistan. The plants were identified botanically, arranged alphabetically along with their Latin name, family name, common name, poisonous parts, toxicity, affects, toxin and their effects. Aim of the study was to induce awareness in the local people of district Bannu about the poisonous effects of the commonly used plants. Data about poisonous effect were collected from the local experienced and mostly old age people through questionnaire. Some information were collected from a number of veterinary texts and literature. The most important plants genera studied in the area were Brassica 6 species (11.11%), Lathyrus 5 spp (9.26%), Astragalus, Euphorbia and Prunus were with 4 spp (7.40%). Datura, Jatropha, Ranunculus, Solanum and Sorghum were with 3 spp (5.56%) while Allium, Amaranthus, Chenopodium, Melilotus and Taxus were with 2 spp (3.70%). These 15 genera contribute 48 species (55.17 %) while the remaining 39 genera have single species each and contribute 44.83% to the total poisonous flora of the research area. Other important poisonous plants were Anagallis arvensis L., Cannabis sativa, Datura stramonium L., D. metel L., Euphorbia species, Heliotropium europaeum, Ipomoea tricolor, Jatropha curcas, Lolium temulentum L., Malus domestica, Mangifera indica L., Medicago sativa L., Melilotus alba Desr., M. officinalis (L.) Lam., Mirabilis jalapa L., Narcissus tazetta, Nicotiana tabacum L., Sorghum halepense (L) Pers., and Xanthium strumarium. It was concluded that the local population had poor knowledge about the poisonous effect of the plants and the present research work was anticipated for use by health care professionals, veterinarians, farmers, homeowners, as well as botanically curious individuals.

Report: Cytotoxic activities of Rosa brunonii, Calligonum polygonoides, Pegnum harmala and Sueda fruticosa extract using brine shrimp.

The present study was carried out to record and evaluate the effect of Rosa brunonii, Calligonum polygonoides, Sueda fruticosa and Pegnum harmala L., extracts on brine shrimp collected during March-June 2013 from different regions of District Bannu. These four plants were medicinal xerophytes and widely distributed throughout Pakistan. Rosa brunonii is commonly used as a hedge plant for gardening. Calligonum polygonoides and Sueda fruticosa are locally used as a fuel, while Pegnum harmala (L.) is the most important multipurpose medicinal xeric plant, which is used for various purposes. All these selected medicinal xerophytes have inhibitory effect on bacterial growth. In this study the effect of different concentration (10-70 µ/ml) were tested on brine shrimp. The results showed that maximum cytotoxic activities were observed in Rosa brunonii (100.0±0.4), Calligonum polygonoides (100.0±0.2) and Pegnum harmala (L.) (90.0±5.2) while Sueda fruticosa (50.0±7.1) has less cytotoxic property. These activities are may be due to the presence of bioactive constituents.

Ramadan fasting increases leniency in judges from Pakistan and India.

We estimate the impact of the Ramadan fasting ritual on criminal sentencing decisions in Pakistan and India from half a century of daily data. We use random case assignment and exogenous variation in fasting intensity during Ramadan due to the rotating Islamic calendar and the geographical latitude of the district courts to document the large effects of Ramadan fasting on decision-making. Our sample comprises roughly a half million cases and 10,000 judges from Pakistan and India. Ritual intensity increases Muslim judges' acquittal rates, lowers their appeal and reversal rates, and does not come at the cost of increased recidivism or heightened outgroup bias. Overall, our results indicate that the Ramadan fasting ritual followed by a billion Muslims worldwide induces more lenient decisions.

Mulching impact of Jatropha curcas L. leaves on soil fertility and yield of wheat under water stress.

In present studies we have evaluated mulching impact of Jatropha curcas leaves on soil health and yield of two wheat (Triticum aestivum L.) varieties Wadan-2017 (rainfed) and Pirsabak-2013 (irrigated) under imposed water stress. Mulch of Jatropha leaves was spread on the soil surface at the rate of 0, 1, 3 and 5 Mg ha-1 after seed germination of wheat. Water stress was imposed by skipping irrigations for one month at anthesis stage of wheat maintaining 40% soil field capacity. We found a significant decline in soil microbial biomass carbon (30.27%), total nitrogen (22.28%) and organic matter content (21.73%) due to imposed water stress in non-mulch plots. However, mulch application at 5 Mg ha-1 significantly improved soil organic matter (38.18%), total nitrogen (37.75%), phenolics content (16.95 mg gallic acid equivalents/g) and soil microbial biomass carbon (26.66%) as compared to non-mulch control. Soil health indicators like soil carbonates, bicarbonates, electrical conductivity, chloride ions and total dissolved salts were decreased by 5 Mg ha-1 mulch application. We noted a decline in yield indicators like spike weight (14.74%), grain spike-1 (7.02%), grain length (3.79%), grain width (3.16%), 1000 grains weight (6.10%), Awn length (9.21%), straw weight (23.53%) and total grain yield (5.98%) of wheat due to imposed water stress. Reduction in yield traits of wheat due to water stress was higher in Pirsabak-2013 than Wadan-2017. Jatropha leaves mulch application at 5 Mg ha-1 significantly minimized the loss in yield traits of wheat crop caused by water stress. Jatropha curcas leaves mulch application at 5 Mg ha-1 is recommended for the successful establishment of wheat crop under water deficit conditions.

Antimicrobial, antioxidant and cytotoxic properties of Chenopodium glaucum L.

We evaluated phytochemical composition, antibacterial, antifungal, anti-oxidant and cytotoxic properties of aqueous (water) and organic extracts (methanol, ethyl acetate and n-hexane) of Chenopodium glaucum. Highest phenolic content 45 mg gallic acid equivalents (GAE)/g d.w was found in aqueous extract followed by ethyl acetate (41mg GAE/g d.w) and methanol extract (34.46 mg GAE/g d.w). Antibacterial potential of aqueous and organic extracts of C. glaucum was examined against Acinetobacter baumannii, Klebsiella pneumoniae, Escherichia coli and Staphylococcus epidermidis. The aqueous, methanolic, ethyl acetate, and n-hexane extract showed antibacterial activity against A. baumannii, K. pneumoniae, E. coli and S. epidermidis. However, against A. baumannii significantly higher inhibition zone (19 mm and 18.96 mm respectively) was shown by ethyl acetate and methanol extracts. Aqueous extract possessed highest growth inhibition (11 mm) against E. coli. Aqueous, ethyl acetate and methanol extracts showed 9 mm, 10 mm, and 10.33 mm zone of inhibition against the K. pneumoniae. For antifungal activity, the extracts were less effective against Aspergillus niger but showed strong antifungal activity against Aspergillus flavus (A. flavus). The antioxidant activity was measured as DPPH (2, 2-diphenyl-1-picrylhydrazyl), H2O2 and ABTS (2, 2'-azino-bis (3-ethylbenzothiazoline-6-sulfonic acid) scavenging activity of free radicals. All the organic extracts of C. glaucum possessed ABTS, DPPH and H2O2 scavenging properties. The highest cytotoxic activity measured as half maximal inhibitory concentration (IC50) against human lungs carcinoma cells was recorded for methanolic (IC50 = 16 µg/mL) and n-hexane (IC50 = 25 µg/mL) extracts, respectively. The Gas chromatography-mass spectrometry (GC-MS) analysis showed 4 major and 26 minor compounds in n-hexane extract and 4 major and 7 minor compounds in methanol extract of the C. glaucum. It is concluded that aqueous and organic extracts of C. glaucum would be potential therapeutic agents and could be exploited on a pilot scale to treat human pathogenic diseases.

Analysis of deformable distortion in the architecture of leaf xylary vessel elements of Carthamus oxycantha caused by heavy metals stress using image registration.

Anatomical study of leaf xylary vessel elements of Carthamus oxycantha under various intensities of lead (Pb) and nickel (Ni) stress (200, 400, 600, and 800 mg Pb(NO3 )2 , NiCl2 ·6H2 O/kg of the soil) was conducted. The deformations caused due to metal stress were detected using point-based image registration technique. Initially, a set of corresponding feature points called landmarks was selected for warping of two-dimensional microscopic images of deformed/source vessel (stressed) to its normal/target (unstressed) counterpart. The results show that the target registration error is less than 3 mm using real plant image datasets. The stress caused alterations mainly in diameter, size, and shape of the cells. Average cell diameter and average wall diameter of vessels were measured with "Image J." The range of decrease in average cell diameter from 18.566 to 13.1 µm and the range of increase in average wall diameter was from 5.166 to 10.1 µm, with increase in stress factor through 200, 400, 600, and 800 mg Pb(NO3 )2 , NiCl2 ·6H2 O/kg of the soil. We noted large deformation in the form of shrinkage in cell size and diminution in its diameter. The diminution in diameter and the shrinkage in cell size of vessel cells may be due to the deposition of wall materials. It can be a possible strategy to limit the water flow to overcome the rapid mobility and transportation of the excess amount of metals to safeguard the cellular components from unpleasant consequences of metallic stress.

Spasmolytic and Ca++ Channel Blocking Potential of Nepetolide: Isolated from Nepeta suavis.

Nepeta suavis is used in traditional medicine for treatment of abdominal spasm (colic). The tricyclic clerodane type diterpene, nepetolide, isolated for the first time from Nepeta suavis, was evaluated for Ca++ antagonist and antispasmodic activities. When studied in isolated rabbit jejunum, nepetolide caused concentration-dependent (0.03-100 µM) relaxation of spontaneous and high K+ (80 mM)-induced contractions, like that caused by verapamil, indicating that nepetolide exhibits spasmolytic activity, possibly mediated through Ca++ channel blocking action, which provides scientific explanation for the medicinal application of Nepeta suavis as an antispasmodic agent.