Launaea fragilis extract attenuated arthritis in rats through modulation of IL-1ß, TNF-α, IL-6, NF-κB, COX-2, IL-4, and IL-10.

Launaea fragilis (Asso) Pau is a Cholistan desert medicinal plant. Launaea species are used as traditional remedies against various inflammatory conditions. The current research was designed to evaluate the anti-nociceptive, anti-inflammatory, and anti-arthritic potential of ethanolic extract of L. fragilis (Et-LF). The plant extract was prepared by triple maceration. GC-MS screening explored the presence of various bioactive phytoconstituents including n-tetracosanol-1, 1-heptacosanol, and n-hexadecanoic acid. DPPH assay demonstrated the antioxidant potential of Et-LF. Safety profile data indicated that Et-LF was safe up to the oral dose of 5000 mg/kg in female rats. Anti-nociceptive activity of Et-LF was assessed in hot plate method and acetic acid-induced writhing model and the results suggested that Et-LF had significant analgesic effects in both animal models. Carrageenan, histamine, and serotonin-induced edema models were used to estimate the anti-inflammatory effects of Et-LF and were found to prevent paw edema development dose dependently. The anti-arthritic effect of Et-LF was estimated in CFA-induced arthritic rat model. Treatment with Et-LF 125, 250, 500 and flurbiprofen (FP) 10 mg/kg/day significantly attenuated the paw edema, reversed the reduced body weight, and restored the altered hematological parameters in arthritic rats. Gene expression studies revealed the significant downregulation of IL-1ß, TNF-α, IL-6, NF­κB, and COX-2, and upregulation of IL-4 and IL-10 in arthritic rats treated with various doses of plant extract. Histological evaluation of ankle joints showed that Et-LF mitigated pannus formation, infiltration of inflammatory cells, and fibrous connective tissue formation in the diseased rats. Thereof, it may be concluded that the recent study demonstrated the anti-nociceptive, anti-inflammatory, and anti-arthritic effects ascribed to the signifying presence of phytoconstituents in L. fragilis.

Dynamics of the time-fractional reaction-diffusion coupled equations in biological and chemical processes.

This paper aims to demonstrate a numerical strategy via finite difference formulations for time fractional reaction-diffusion models which are ubiquitous in chemical and biological phenomena. The time-fractional derivative is considered in the Caputo sense for both linear and nonlinear problems. First, the Caputo derivative is replaced with a quadrature formula, then an implicit method is used for the remaining part. In the linear case, the proposed strategy reduces the time fractional models into linear simultaneous equations. In nonlinear cases, Quasilinearization is utilized to tackle the nonlinear parts. With this strategy, solutions of the fractional system transform into linear algebraic systems which are easy to solve. Next, the Von Neumann method is implemented to examine the stability of the scheme which discloses that the scheme is unconditionally stable. Further, the applicability of the presented scheme is tested with different linear and nonlinear models which include the one dimensional Schnakenberg and Gray-Scott models, and one and two dimensional Brusselator models. To analyze the accuracy of the present technique two norms namely, L ∞ and L 2 , and relative error are addressed. Moreover, the obtained outcomes are shown tabulated and graphically which identifies that the scheme properly works for the time fractional reaction-diffusion systems.

Synthesis of Silver Nanoparticles from Ganoderma Species and Their Activity against Multi Drug Resistant Pathogens.

The widespread and indiscriminate use of broad-spectrum antibiotics leads to microbial resistance, which causes major problems in the treatment of infectious diseases. However, advances in nanotechnology using mushrooms have opened up new domains for the synthesis and use of nanoparticles against multidrug-resistant pathogens. Mushooms have recently attracted attention and are exploited for food and medicinal purposes. The current study focuses on the molecular identification, characterization of biologically synthesized silver nanoparticles by X-ray diffraction (XRD) spectroscopy, Fourier Transform Infrared Spectroscopy (FTIR), UV-Vis spectroscopy and scanning electron microscopy (SEM) and antibacterial analysis of extract and silver nanoparticles (AgNPs) synthesis from Ganoderma resinaceum against multidrug resistant microbes. Accurate identification of mushrooms is key in utilizing them for the benefit of humans. However, morphological identification of mushrooms is time consuming, tedious and may be prone to error. Molecular techniques are quick and reliable tools that are useful in mushroom taxonomy. Blast results showed that G. resinaceum (GU451247) obtained from Pakistan was 97 % same to the recognized G. resinaceum (GU451247) obtained from China as well as G. resinaceum (GU451247) obtained from India. The antimicrobial potential of mushroom composite and AgNPs showed high efficacy against pathogenic Staphylococcus aureus (ZOI 23 mm) K. pneumonia (ZOI 20 mm), Pseudomonas aeruginosa (ZOI 24 mm) and E. fecalis and A. baumannii (ZOI 10 mm), and multidrug resistant (MDR) A. baumannii (ZOI 24 mm). XRD evaluation revealed the crystalline composition of synthesized NPs with diameter of 45 nm. UV-Vis spectroscopy obsorption peaked of 589 nm confirmed the presence of AgNPs. SEM results showed the cubic morphology of AgNPs. The FTIR analysis of NPs obtained from G. resinaceum containing C=O as well as (O=C-H) stretching revealed presence of hydrogen, carbonyl and amide groups. The synthesized extract and AgNPs showed promising minimum inhibitory concentration (MIC) at 2 mg concentration against the MDR strains. AgNPs are observed to be efficient as they need less quantities to prevent bacterial growth. In the view of challenges for developing antimicrobial NPs of variable shape and size by various other methods, tuning nanoparticles synthesized via mushrooms can be a wonderful approach to resolve existing hurdles.

Molecular Profiling, Characterization and Antimicrobial Efficacy of Silver Nanoparticles Synthesized from Calvatia gigantea and Mycena leaiana against Multidrug-Resistant Pathogens.

The use of natural products isolated from mushrooms against infection, cancer diseases and other oxidative-stress-related diseases is one of the cornerstones of modern medicine. Therefore, we tried to establish a combination of medicinal mushrooms and nanotechnology possibly with the field of medicine for the development of antibacterial agents against these MDR strains. The aim of the research was to understand the molecular identification, characterization and antibacterial action of Calvatia gigantea and Mycena leaiana. The identification of fruiting body species via morpho-anatomical and molecular methods was necessary to analyze the genetic variability and phylogenetic relationships of mushrooms. Phylogenetic analysis revealed that Calvatia from Hunza, Pakistan, exhibited 98% resemblance to the previously discovered Langermannia gigantean (DQ112623) and L. gigantean (LN714562) from northern Europe, and Mycena (Pakistan) showed a 97% similarity to M. leaiana (MF686520) and M. leaiana (MW448623) from the USA. UV-vis, scanning electron microscopy (SEM), Fourier transform infrared spectroscopy (FTIR), and X-ray diffraction (XRD) were used for AgNPs' characterization. The UV-vis absorption peak of 500-600 nm indicates the AgNPs' presence. XRD results determined Calvatia gigantea AgNPs were nanocrystals and Mycena leaiana seems to be amorphous. In addition, SEM results showed the cubic morphology of C. gigantea with a diameter of 65 nm, and the FTIR spectra of fruiting body revealed the presence of functional groups-carboxyl, nitro, and hydroxyl-in AgNPs, which catalyzed the reduction of Ag+ to Ag0. Further antibacterial activity of mushrooms against MDR strains was determined via agar well diffusion assay, and Minimum Inhibitory Concentration (MIC) was estimated by qualitative experimentation using the broth dilution method. All experiments were conducted in triplicate. The results showed that the mushroom AgNPs, along with their synergy and nano-composites (with the exception of Ethyl-acetate), were shown to have zones of inhibition from 4 mm to 29 mm against multidrug-resistant pathogens such as Acinetobacter baumannii, Staphylococcus aureus, Pseudomonas aeruginosa, Klebsiella pneumonia, Proteus mirabilis, Enterobacter cloacae and Escherichia coli. The mushroom composites were active against most of the tested microorganisms whilst the lowest MIC value (10-40 mg/mL) was recorded against MDR strains. Hence, the present study suggested the possibility of employing compounds present in mushrooms for the development of new antibacterial agents, as well as efflux pump inhibitors.

A facile two-step hydrothermal preparation of 2D/2D heterostructure of Bi2WO6/WS2 for the efficient photodegradation of methylene blue under sunlight.

A facile two-step hydrothermal method was successfully used to prepare a photocatalyst Bi2WO6/WS2 heterojunction for methyl blue (MB) photodegradation. Fabricated photocatalysts were characterized by X-ray diffraction (XRD), scanning electron microscope (SEM), energy-dispersive X-ray analysis (EDX), and X-ray photoelectron spectroscopy (XPS). Band gap measurements were carried out by diffuse reflectance spectroscopy (DRS). Results indicated that the prepared heterostructure photocatalyst has increased visible light absorption. Photocatalytic performance was evaluated under sunlight irradiation for methylene blue (MB) degradation as a model dye. Variations in pH (4-10), amount of catalyst (0.025-0.1 g/L), and initial MB concentrations (5-20 ppm) were carried out, whereas all prepared catalysts were used to conduct the tests with a visible spectrophotometer. Degradation activity improved with the pH increase; the optimum pH was approximately 8. Catalyst concentration is directly related to degradation efficiency and reached 93.56% with 0.075 g of the catalyst. Among tested catalysts, 0.01 Bi2WO6/WS2 has exhibited the highest activity and a degradation efficiency of 99.0% in 40 min (min) for MB. MB photodegradation follows pseudo-first-order kinetics, and obtained values of kapp were 0.0482 min-1, 0.0337 min-1, 0.0205 min-1, and 0.0087 min-1 for initial concentrations of 5 ppm, 10 ppm, 15 ppm, and 20 ppm, respectively. The catalyst was reused for six cycles with a negligible decrease in the degradation activity. Heterostructure 0.01 Bi2WO6/WS2 has exhibited a photocurrent density of 16 µA cm-2, significantly higher than 2.0 and 4.5 µA cm-2 for the pristine WS2 and Bi2WO6, respectively. The findings from these investigations may serve as a crucial stepping stone towards the remediation of polluted water facilitated by implementing such highly efficient photocatalysts.

The fungicide azoxystrobin causes histopathological and cytotoxic changes in the midgut of the honey bee Apis mellifera (Hymenoptera: Apidae).

Apis mellifera is an important bee pollinating native and crop plants but its recent population decline has been linked to the use of pesticides, including fungicides that have been commonly classified as safe for bees. However, many pesticides, in addition to direct mortality cause sublethal effects, including damage to target selective honey bee organs. The midgut is the organ responsible for the digestion and absorption of nutrients and the detoxification of ingested substances, such as pesticides. This study evaluated the histopathological and cytotoxic changes in the midgut of A. mellifera workers caused by the pesticide azoxystrobin. The limit-test was performed, and a 100 µg a.i./bee dose was administered orally and midgut analyzed with light and transmission electron microscopies after 24 h and 48 h of pesticide exposure. The midgut of the control bees has a single layer of digestive cells, with spherical nuclei, nests of regenerative cells, and the lumen coated with the peritrophic matrix. The bees fed on azoxystrobin showed morphological changes, including intense cytoplasm vacuolization and cell fragments released into the gut lumen. The protein detection test showed greater staining intensity in the nests of regenerative cells after 24 h of exposure to azoxystrobin. The occurrence of damage to the midgut in A. mellifera exposed to azoxystrobin indicates that although this fungicide has been classified as low toxicity for bees, it has sublethal effects in the midgut, and effects in other organs should be investigated.

Development of efficient bi-functional g-C3N4@MOF heterojunctions for water splitting.

Herein we report the development of highly efficient heterojunctions by combining n-type g-C3N4 and MOFs as bi-functional photoelectrocatalysts towards the hydrogen evolution reaction (HER) and the oxygen evolution reaction (OER). g-C3N4@MIL-125(Ti) and g-C3N4@UiO-66 have been synthesized via in situ incorporation of pre-synthesized g-C3N4 nanoparticles into MIL-125(Ti) and UiO-66. Bare MIL-125(Ti) and UiO-66 are also prepared for comparison. All the synthesized samples have been characterized by Powder X-ray Diffraction analysis, Fourier Transform Infrared Spectroscopic analysis, Scanning Electron Microscopic analysis, Energy Dispersive X-ray Spectrometry and UV-Vis Spectroscopic analysis. Cyclic voltammetry and linear sweep voltammetry studies have been carried out for all samples which indicates that under visible light exposure the g-C3N4@MIL-125(Ti)/NF heterojunction achieved a current density of 10 mA cm-2 at just 86 and 173 mV overpotential for the HER and OER, respectively. Moreover, all the synthesized samples display significant stability and generate a constant current density up to 1000 cyles during water electrolysis performed at a constant applied potential 1.5 V.

Ultrasound-mediated blood-brain barrier opening: An effective drug delivery system for theranostics of brain diseases.

Blood-brain barrier (BBB) remains a significant obstacle to drug therapy for brain diseases. Focused ultrasound (FUS) combined with microbubbles (MBs) can locally and transiently open the BBB, providing a potential strategy for drug delivery across the BBB into the brain. Nowadays, taking advantage of this technology, many therapeutic agents, such as antibodies, growth factors, and nanomedicine formulations, are intensively investigated across the BBB into specific brain regions for the treatment of various brain diseases. Several preliminary clinical trials also have demonstrated its safety and good tolerance in patients. This review gives an overview of the basic mechanisms, ultrasound contrast agents, evaluation or monitoring methods, and medical applications of FUS-mediated BBB opening in glioblastoma, Alzheimer's disease, and Parkinson's disease.

Risk variants of obesity associated genes demonstrate BMI raising effect in a large cohort.

Obesity is highly polygenic disease where several genetic variants have been reportedly associated with obesity in different ethnicities of the world. In the current study, we identified the obesity risk or protective association and BMI raising effect of the minor allele of adiponectin, C1Q and collagen domain containing (ADIPOQ), cholesteryl ester transfer protein (CEPT), FTO alpha-ketoglutarate dependent dioxygenase (FTO), leptin (LEP), and leptin receptor (LEPR) genes in a large cohort stratified into four BMI-based body weight categories i.e., normal weight, lean, over-weight, and obese. Based on selected candidate genetic markers, the genotyping of all study subjects was performed by PCR assays, and genotypes and allele frequencies were calculated. The minor allele frequencies (MAFs) of all genetic markers were computed for total and BMI-based body weight categories and compared with MAFs of global and South Asian (SAS) populations. Genetic associations of variants with obesity risk were calculated and BMI raising effect per copy of the minor allele were estimated. The genetic variants with higher MAFs in obese BMI group were; rs2241766 (G = 0.43), rs17817449 (G = 0.54), rs9939609 (A = 0.51), rs1421085 (C = 0.53), rs1558902 (A = 0.63), and rs1137101 (G = 0.64) respectively. All these variants were significantly associated with obesity (OR = 1.03-4.42) and showed a high BMI raising effect (ß = 0.239-0.31 Kg/m2) per copy of the risk allele. In contrast, the MAFs of three variants were higher in lean-normal BMI groups; rs3764261 A = 0.38, rs9941349 T = 0.43, and rs7799039 G = 0.40-0.43). These variants showed obesity protective associations (OR = 0.68-0.76), and a BMI lowering effect per copy of the protective allele (ß = -0.103-0.155 Kg/m2). The rs3764261 variant also showed significant and positive association with lean body mass (OR = 2.38, CI = 1.30-4.34). Overall, we report six genetic variants of ADIPOQ, FTO and LEPR genes as obesity-risk markers and a CETP gene variant as lean mass/obesity protective marker in studied Pakistani cohort.

Gazing the dusty mirror: Joint effect of narcissism and sadism on workplace incivility via indirect effect of paranoia, antagonism, and emotional intelligence.

Workplace productivity is badly affected by many negative factors such as narcissism, and sadism. In addition, paranoia and antagonism play an important role in increasing workplace incivility. Through emotional intelligence, such negative behaviors could be addressed by managers and their junior colleagues. The current study aims to investigate the parallel mediating role of paranoia, antagonism, and emotional intelligence on the relationship between narcissism, sadism, and workplace incivility. A survey approach was used. Primary data was collected in PLS-SEM. The population of the study was all faculty members in higher education institutions in the Khyber Pakhtunkhwa (Pakistan) region. A measurement model and structural model were developed. The measurement model demonstrated that convergent and discriminant validities were established. The structural model's findings revealed that narcissism, antagonism, and emotional intelligence were not mediated between narcissism and workplace incivility. Similarly, emotional intelligence did not play any mediating role between sadism and workplace incivility. This implied that emotional intelligence has no role in decreasing or reducing workplace uncivil behavior.

Impact of Dark Triad on Anxiety Disorder: Parallel Mediation Analysis During Pandemic.

The current study aimed to investigate the mediating role of loneliness, ASMR, on the relationship between narcissism, Machiavellianism, and psychopathy and anxiety disorder. The population of the study was professionals working in public and private sector organizations. Non-probability snowball technique was used. Data was collected from 512 professionals. A total of 653 questionnaires were distributed and 512 were used in the analysis yielding a response rate of 78.4%. PLS-SEM was used to analyze the data. Measurement and structural models were developed to test the reliability, and validity of the scales as well as hypotheses. Findings of the measurement model revealed that scales were reliable and valid while results of the structural model revealed that narcissism, psychopathy, COVID loneliness, and ASMR have a significant direct impact on anxiety disorder but Machiavellianism does not have a significant effect on anxiety disorder. In addition, COVID loneliness and ASMR mediated between narcissism and psychopathy but do not significantly mediate between Machiavellianism and anxiety disorder. The current study has extended the body of knowledge by bridging the two theories theory of attachment and cognitive dissonance theory. The current study has provided the primary evidence that COVID loneliness increases anxiety while ASMR (audio-visual) tingling sensations help to reduce anxiety disorder.

The Moderating Role of Organizational Citizenship Behavior Toward Environment on Relationship Between Green Supply Chain Management Practices and Sustainable Performance.

The purpose of this study was to investigate the moderating role of "organizational citizenship behavior toward the environment" (OCBE) on the relation between "green supply chain management" (GSCM) practices and sustainable performance. The participants of the current study were manufacturing firms, and non-probability convenience sampling technique was used for selecting the sample size. The survey method used while data were collected from manufacturing firms was cross-sectional; a total of 151 responses were received and used in the analysis. For statistical analysis, "SmartPLS partial least square, structural equation modeling" PLS-SEM was used. From the findings, it was evident that in the measurement model, convergent and discriminant validities were established. In the second stage, a structural model was developed for hypotheses testing. It was found that sustainable performance was associated with proposed GSCM practices, excluding environmental management. OCBE being a moderator has significant impacts concerning GSCM operations and sustainability functions of firms. However, OCBE did not play the role of moderator between internal environmental management and EE and sustainable performance. The present study is significant for managers and policymakers for the attainment of competitive advantage, enhancement of OCBE, and sustainable performance. Furthermore, this research study is the first empirical study that has used OCBE as a moderator through the lens of NRBV.

Ultrasound imaging tracking of mesenchymal stem cells intracellularly labeled with biosynthetic gas vesicles for treatment of rheumatoid arthritis.

Rationale: Rheumatoid arthritis (RA) is an autoimmune disease characterized by chronic inflammation and damage to articular tissues that can lead to irreversible joint damage and progressive disability. The multipotent mesenchymal stem cells (MSCs) play an important role in immune disorders and tissue regeneration. However, their immunosuppressive effects and the underlying mechanisms are largely unclear due to the lack of tools for real-time imaging of MSCs in vivo. Gas vesicles (GVs) are biosynthetic nanobubbles that are ejected from aquatic microbes, such as bacteria and archaea, and have an excellent ultrasound imaging capacity. Methods: We harvested MSCs from the bone marrow of Sprague Dawley (SD) rats. Then, GVs were synthesized and incubated with MSCs to obtain intracellularly labeled MSCs. We firstly tested the ultrasound imaging of GV@MSCs in vitro and in vivo and then explored the therapeutic effect of GV@MSCs combined with methotrexate (MTX) in RA rats. Results: These GV@MSCs showed significant contrast-enhanced ultrasound signals without a loss of viability and differentiation capacity. In addition, the GV@MSCs could be imaged in real-time for 5 days using ultrasound both in vitro and in vivo, making it possible to visually track their migration and homing to the joint cavity from the subcutaneous layer of lateral malleolus joints in the injected RA rats. Furthermore, GV@MSCs significantly enhanced the curative effect of methotrexate (MTX) against RA, resulting in decreased paw thickness, lower arthritis index score, reduced bone erosion and cartilage destruction, compared to the PBS, free MTX, and GV@MSCs groups. Conclusion: We developed a novel therapeutic strategy against RA using GVs-loaded MSCs that can be tracked in vivo in real-time.

Molecular Evaluation of Kyoho Grape Leaf and Berry Characteristics Influenced by Different NPK Fertilizers.

Fertilization, a fundamental aspect of a plant's life, has been of great concern for agricultural specialists to minimize the yield gap between actual and potential yield. Around the globe, fertilizers with different NPK ratios are being used to attain a better yield of grape. To find the suitable commercially available fertilizer for quality grape production, a 2 years (2017-2018) study was conducted for the evaluation of 10 fertilizers with different NPK ratios. Commercial fertilizers included were Zhanlan (16:16:16), Garsoni (15:15:15), Acron (16:16:16), Norway (21:7:12), Peters 1 (30:10:10), Nutrivant (14:14:30), Peters 2 (20:20:20), UMAX (15:15:15), G2 (20:20:20), and Yara (15:15:15). The fertilizer application rate was 20 g plant-1, and each was applied at L-29, L-33, and L-36 phenological stages. Chlorophylls, carotenoids, macro/micronutrients in leaf, and anthocyanin derivatives in grape peel were evaluated. Expression levels of 24 genes, including nitrogen, phosphorous, potassium, and anthocyanin pathways in leaf, peel, and pulp were validated by qPCR at L-29, L-33, and L-36 stages. Results indicated that Norway (21:7:12) and Peters 1 (30:10:10) increased carotenoids, chlorophylls, and anthocyanins in leaves, while Zhanlan (16:16:16) improved fruit biochemical attributes, and anthocyanin (cyanidin, delphinidin, petunidin, malvidin, peonidin, and pelargonidin contents). However, a better grape yield was obtained by the application of Peters 1 (30:10:10). Potassium pathway genes were upregulated by Nutrivant (14:14:30), phosphorous pathway genes by Peters 2 (20:20:20), and nitrogen pathway genes by Peters 1 (30:10:10), while Nutrivant (14:14:30) upregulated anthocyanin pathway genes and simultaneously enhanced anthocyanin biosynthesis in berry peels. Results of two years' study concluded that Peters 1 (30:10:10) was proved better to increase yield, while Zhanlan (14:14:30) was superior in improving anthocyanin biosynthesis.

Identification of GH17 gene family in Vitis vinifera and expression analysis of GH17 under various adversities.

Glycoside hydrolase (GH, EC 3.2.1) is a group of enzymes that hydrolyzes glycosidic bonds and play a role in the hydrolysis and synthesis of sugars in living organisms. Vitis vinifera is an important fruit crop and it harbors GH17 gene family however, their function in grapes has not been systematically investigated. In this study, a total of 870 GH17 genes were identified from 14 plant species and their structural domain, sequence alignment, phylogenetic tree, collinear analysis, with the expression profiles of VviGH17 gene family was performed. The promoter analysis of VviGH17 gene showed the presence of cis-acting elements, which are responsive to plant growth and development. In addition, elements for plant hormones were found that are triggered in response to abiotic/biological stress. Transcriptomic data led to the identification of several VviGH17 genes, which are associated with bud dormancy and in response to abiotic stress. Transcript analysis was carried out for some of the selected VviGH17 genes RT-qPCR. VviGH17-16 and VviGH17-30 genes were differentially expressed during bud dormancy, fruit development and different abiotic stresses. Moreover, VviGH17-37 and VviGH17-44 were differentially expressed at fruit development, in response to abiotic stress. In addition, subcellular localization predicts that the VviGH17-16, VviGH17-30, and VviGH17-37 genes were located in the cell membrane, while VviGH17-44 gene was located in the vacuole. In conclusion, our study led to the identification of several GH17s and their probable role in development and stress. SUPPLEMENTARY INFORMATION: The online version contains supplementary material available at 10.1007/s12298-021-01014-1.

Behavioral and ultrastructural effects of novaluron on Aedes aegypti larvae.

Chitin synthesis inhibitors (CSI) are supposed to inhibit formation of chitin microfibrils in newly synthesized cuticle during molting process. Conversely, there has been comparatively few data on morphological effects of CSI on non-target insect organs. In this work, the effects of the CSI novaluron on behavior and midgut of A. aegypti were evaluated. Toxicity bioassays revealed that novaluron is toxic to A. aegypti larva with LC50 = 18.57 mg L-1 when exposed in aqueous solution for 24 h. Novaluron treated larvae were less active and spent more time resting compared to the control group. Histopathology showed that midguts of novaluron-treated larvae had cytoplasm vacuolization and damaged brush border. Cytotoxic effects in midguts of treated larvae induced necrosis, autophagy and damage to mitochondria. Despite being chitin synthesis inhibitor, novaluron did not induce alterations in the integument of A. aegypti larvae. Fluorescence microscopy revealed that the number of digestive cells were higher in novaluron-treated larvae than in control, in response to digestive cell apoptosis. The present study highlights the importance of novaluron against A. aegypti larvae by causing injuries to non-target organs, altering behaviors, inducing cell death and inhibiting cell proliferation.

First Report of the Rust Disease Caused by Puccinia crepidis-japonicae on New Host Sonchus arvensis from Pakistan.

Sonchus arvensis (Asteraceae) is a traditional medicinal herb. The aerial parts are a rich source of vitamins, essential amino acids, and minerals, which may help in treatment of fever, inflammation, detoxication, and blood circulation (Li et al. 2018). In October 2018, typical rust symptoms were observed on S. arvensis leaves and stems in Buner district, Malakand division, Khyber PakhtunKhwa Province, Pakistan (34.39°N; 72.61°E). Almost 40% of leaves and stems of five S. arvensis plants displayed severe rust. The specimens were examined microscopically and compared with type specimen description in published literature (Dietel 1908; Hiratsuka et al. 1992). The fungus was identified as Puccinia crepidis-japonicae based on the characteristic of spore morphology and phylogenetic study based on the internal transcribed spacer (ITS) and large subunit (LSU) sequence data. Uredinia were amphigenous, rounded, or somewhat elliptical, naked, small patches, equally spread, brown. Urediniospores (n=30) were globose to ellipsoid, brownish yellow and measured 22.4-24.7 × 20.2-22.1 µm. Urediniospore walls were brownish orange and finely echinulate and 1.7 to 2.1 µm thick with 2 to 3 germ pores. Telia were amphigenous, rounded or elliptic, scattered, dark brown to blackish. Teliospores (n=30) were ellipsoid, subglobose or long ellipsoid, rounded at both ends, not thickened at apex, warted, reddish brown, and measured 31.3-39 × 24.6-26.8 µm. Teliospore walls were reddish-black and about 1.5-2.5 µm thick, and the pedicles were short, hyaline, fragile, become tapered toward apex, and measured 14.4-18.7 × 4.7-9 µm. DNA was extracted from urediniospores, and the combined region of ITS and LSU (28S) were amplified using Rust2Inv (forward primer) and LR6 (Reverse primer) according to the protocol outlined by Aime (2006). A BLASTn search (http://www.ncbi.nlm.nih.gov) showed that the combined ITS and LSU region shared 99% identity (792/804 bp) to the P. crepidis-japonicae accessions (KY798395 from Hawaii, USA) with 100% query cover. The resulting sequence was deposited in GenBank (Accession No. MN093335). Both morphological and molecular characteristics indicatethat this species was P. crepidis-japonicae. To test pathogenicity and fulfill the Koch's postulates, a urediniospore suspension (1 × 104 spores/ml) was sprayed on three 6-week-old plants of S. arvensis, and one as negative control, incubated at 22-24°C. Uredinia were observed on the leaves after 10 days of inoculation, whereas the control plants remained symptomless. Microscopic examination confirmed that the symptoms on plants obtained from the field and greenhouse inoculations were morphologically identical. This fungus has been observed previously on Crepis japonica in China, Hong Kong, Japan, Korea, and Taiwan, on Prenanthes spp. in China and on Youngia tenuifolia and Y. fusca in China (Farr and Rossman 2021). Pereira et al. (2002) suggested that P. crepidis-japonicae may play a significant role as a biocontrol agent against its weed host. To the best of our knowledge, there are no other reports of this fungus on any other hosts in Pakistan. The specimen has been vouchered in LAH Herbarium, Department of Botany, University of the Punjab, Lahore, Pakistan (LAH36343). This is the first report of P. crepidis-japonicae on S. arvensis as a new host from Pakistan.

Iron and Manganese Codoped Cobalt Tungstates Co1-(x+y)Fe x Mn y WO4 as Efficient Photoelectrocatalysts for Oxygen Evolution Reaction.

Photoelectrocatalysts are robust materials for the production of energy through different ways such as water splitting. Narrow optical band gaps and high overpotentials are limiting the development of photoelectrocatalysts. In this study, a series of Co1-(x+y)Fe x Mn y WO4 solid solutions of cobalt tungstate codoped with iron and manganese have been synthesized hydrothermally. The synthesized solid solutions have been characterized by powder XRD, UV-visible spectra, cyclic voltammetry (CV), and linear sweep voltammetry (LSV). They all crystallize in a wolframite-type monoclinic crystal system with space group P2/c. Doping of iron and manganese leads to narrowing of the optical band gap of Co1-(x+y)Fe x Mn y WO4 from 2.60 to 2.04 eV. The electrocatalytic activity toward oxygen evolution reaction of all of the samples has been evaluated through LSV measurements. It is found that the sample named C5, which is codoped with manganese and iron, has the lowest onset potential and needs the lowest overpotential to attain the targeted 5 mA cm-2 and standard 10 mA cm-2 current densities as compared with all other synthesized samples. This study shows that the synthesized tungstates can be good candidates for the photoelectrocatalytic oxygen evolution reaction.

The salivary glands of Brontocoris tabidus (Heteroptera: Pentatomidae): Morphology and secretory cycle.

Brontocoris tabidus (Signoret) (Heteroptera: Pentatomidae) is a zoophytophagous insect used for biological control in agriculture and forest systems because its nymphs and adults feed on insects and plants. The predatory Pentatomidae insert the mouthparts into the prey, releasing saliva to paralysis and kills the insect, as well as digest body parts to be sucked in a preliminary extra-oral digestion. In a short period of time, this insect shows the ability to feed again, suggesting the existence of a constant and abundant secretory cycle in the salivary glands. This study evaluated the morphological, histochemical and ultrastructural changes of the salivary glands of B. tabidus in fed and starved insects. The salivary complex of this predatory bug has a pair of bilobed salivary glands and a pair of tubular accessory salivary glands. The accessory glands have the lumen lined by a thick non-cuticular layer rich in glycoproteins. The secretory cells of the B. tabidus principal salivary glands have constant secretory activity, with each lobe producing different substances. The physiological processes that occur in the salivary gland of B. tabidus indicate that the insect needs to feed constantly, corroborating the potential of this insect to be used in biological control programs.

Exposure to chlorantraniliprole reduces locomotion, respiration, and causes histological changes in the midgut of velvetbean caterpillar Anticarsia gemmatalis (Lepidoptera: Noctuidae).

The anthranilic diamide, chlorantraniliprole is a systemic insecticide affecting ryanodine receptors. This insecticide is used to control caterpillars in soybean crops because it has low toxicity to non-target organisms. The objective was to identify side-effects of chlorantraniliprole on midgut histopathology, respiration and behavior of the velvetbean caterpillar Anticarsia gemmatalis in laboratoty. Chlorantraniliprole has LC50 = 0.61 (0.58-0.64) mg mL-1 for A. gemmatalis fourth instar caterpillars after 96 h. The insecticide causes severe histopathological effects in the midgut with epithelial disorganization, microvilli degeneration, cytoplasm vacuolization, cell fragmentation, and peritrophic matrix disorganization. The respiratory rate and the walking speed decrease, whereas the resting period increase for caterpillars exposed to this insecticide. Chlorantraniliprole is toxic to A. gemmatalis at median lethal concentrations causing severe histological and ultrastructural changes with degeneration of the midgut epithelium, reduction of respiratory rates and inducing an arresting behavioral response of this insect.