Herbicidal proteins from Bacillus wiedmannii isolate ZT selectively inhibit ryegrass (Lolium temulentum L.).

BACKGROUND: The widespread use of chemical herbicides and the growing issue of weed resistance pose significant challenges in agriculture. To address these problems, there is a pressing need to develop biological herbicides based on bacterial metabolites. RESULTS: In this study, we investigated the impact of the cell-free culture filtrate (CFCF) from the ZT isolate, a bacilliform bacterium obtained from diseased wheat seeds, on the germination and seedling growth of various plant species, including wild oat, ryegrass, redroot, wheat, and chickpea. The results revealed that CFCF had a detrimental effect on the fresh and dry weight of stems and roots in most of the studied plants, except chickpeas. The CFCF was further subjected to separation into aqueous and organic phases using chloroform, followed by the division of the aqueous phase into 13 fractions using an alumina column. Notably, both the aqueous phase (20%) and all 13 fractions (ranging from 50% to 83%) displayed the ability to reduce the root length of ryegrass, a monocotyledonous weed. Liquid chromatography-mass spectrometry (LC-MS) analysis identified that fractions 3 and 7, which were effective against ryegrass but not redroot, contained Cry family proteins, including Cry10 Aa, Cry4 Ba, and Cry4 Aa. Additionally, 16s rRNA gene sequencing revealed that the ZT isolate is closely related (98.27%) to Bacillus wiedmannii. CONCLUSION: Conclusively, metabolites from the ZT bacterium hold promise for monocotyledonous weed-targeted herbicides, providing a constructive strategy to confront agricultural issues tied to chemical herbicides and weed resistance. © 2024 Society of Chemical Industry.

The effects of induced type I diabetes on developmental regulation of GDNF, NRTN, and NCAM proteins in the dentate gyrus of male rat offspring.

BACKGROUND: Maternal diabetes during pregnancy can affect the neurological development of offspring. Glial cell-derived neurotrophic factor (GDNF), neurturin (NRTN), and neural cell adhesion molecules (NCAM) are three important proteins for brain development. Therefore, this study aimed to investigate the impacts of the mentioned neurotrophic factors in the hippocampal dentate gyrus (DG) of rat offspring born to diabetic mothers. METHODS: Wistar female rats were randomly allocated into diabetic (STZ-D) [(45 mg/kg BW, STZ (Streptozotocin), i.p)], diabetic + NPH insulin (STZ-INS) [(4-6 unit/kg/day SC)], and control groups. The animals in all groups were mated by non-diabetic male rats. Two weeks after birth, male pups from each group were sacrificed and then protein contents of GDNF, NRTN, and NCAM were evaluated using immunohistochemistry. RESULTS: The study found that the expression of GDNF and NRTN in the hippocampus of diabetic rat offspring was significantly higher compared to the diabetic+ insulin and control groups, respectively (P < 0.01, P < 0.001). Additionally, the expression of NCAM was significantly higher in the diabetic group the diabetic+ insulin and control groups (P < 0.01, P < 0.001). CONCLUSIONS: The results of the study revealed that diabetes during pregnancy significantly impacts the distribution pattern of GDNF, NRTN, and NCAM in the hippocampus of rat neonates.

The antidepressant effect of combined extracts of Hypericum perforatum and Echium amoenum supplementation in patients with depression symptoms: A randomized clinical trial.

Objective: Echium amoenum and Hypericum perforatum dried flowers have been used for therapy of mental disorders in Iranian traditional medicine. In this study, we assessed the efficacy of the E. amoenum and H. perforatum extracts in patients with mild to moderate depression. Materials and Methods: In an 8-week double-blind, parallel-group trial, 51 patients randomly consumed 20 mg of fluoxetine or 350 mg of herbal medicine twice daily. The Hamilton Rating Scale for Depression (HAM-D) was used to assess depression severity in patients at weeks 0, 4, and 8. Results: According to the Hamilton score, there were no significant differences between the fluoxetine- and herbal medicine-treated groups after 4 and 8 weeks (p>0.05). Dry mouth was the only reported side effect which was significantly lower in the herbal group (p<0.05) in weeks 2 and 4. Conclusion: E. amoenum and H. perforatum have anti-depressant properties similar to fluoxetine and they can be used to treat depression as an alternative to fluoxetine.

Specific Streptomyces strain enhances the growth, defensive mechanism, and fruit quality of cucumber by minimizing its fertilizer consumption.

BACKGROUND: The required amounts of chemical fertilizers (NPK) are determined by plant yield, and product quality is given less consideration. The use of PGPRs is an environmentally friendly approach that, in addition to increasing yield, also improves fruit quality. This study examined the role of specific Streptomyces strains in aiding cucumber plants to 1) use fewer NPK fertilizers in the same quantity 2) improve the quality of cucumber fruit, and 3) promote growth and defense system. RESULTS: In this study, the effect of 17 Streptomyces strains on the vegetative traits of cucumber seedlings of the Sultan cultivar was evaluated as the first test. Four strains of Streptomyces with the highest root and shoot dry weight were selected from the strains. This experiment was performed to determine the interaction effect of selected strains and different amounts of NPK on cucumber yield, quality, physiological and biochemical responses of plants. The first experiment's results revealed that strains IC6, Y7, SS12, and SS14 increased significantly in all traits compared to the control, while the other strains dramatically improved several characteristics. Analysis of variance (ANOVA) revealed significant differences between the effect of strains, NPK concentrations, and their interactions on plant traits. The treatments containing 75% NPK + SS12, yielded the most fruit (40% more than the inoculated control). Antioxidant enzymes assay showed that SS12 substantially increased the activity of POX, PPO, and the expression of the genes related to these two enzymes. Hormone assay utilizing HPLC analysis revealed that various strains employ a specific mechanism to improve the immune system of plants. CONCLUSIONS: Treatment with strain SS12 led to the production of cucumbers with the highest quality by reducing the amount of nitrate, and soluble sugars and increasing the amount of antioxidants and firmness compared to other treatments. A specific Streptomyces strain could reduce 25% of NPK fertilizer during the vegetative and reproductive growth period. Moreover, this strain protected plants against possible pathogens and adverse environmental factors through the ISR and SAR systems.

Antibacterial Effect of Allium ampeloprasum and Allium porrum Extracts on Staphylococcus aureus and Pseudomonas aeruginosa.

Objectives: The most prominent microorganisms that cause hospital infections and acquire antibiotic resistance are Staphylococcus aureus and Pseudomonas aeruginosa. The present study aimed to compare the phenolic and flavonoid compounds of various Allium ampeloprasum and Allium porrum extracts and evaluate the antibacterial effects of these extracts against these two microorganisms. Methods: The total phenolic and flavonoid contents of the acetone, methanol, aqueous, and hexane leeks extracts from A. ampeloprasum and A. porrum were measured. The antibacterial activity of these extracts against S. aureus and P. aeruginosa was tested using the disk diffusion method for 24, 48, and 72 hours. Further, the minimum inhibitory concentrations and the minimum bactericidal concentrations of these extracts for these two bacteria were evaluated and compared with those of common antibiotics. Results: The aqueous extracts showed the highest phenolic and flavonoid contents and at concentrations of 35 and 40 mg per disk, showed the most antibacterial activity against S. aureus and P. aeruginosa; P. aeruginosa showed more sensitivity to the aqueous extracts than S. aureus. Conclusion: Aqueous A. ampeloprasum and A. porrum extracts may prevent the growth of hospital pathogens, especially P. aeruginosa; our findings will aid the discovery of new antimicrobial substances against antibiotic-resistant bacteria.

Streptomyces strains can improve the quality properties and antifungal bioactivities of tomato fruits by impacting WRKY70 transcription factor gene and nitrate accumulation.

The current study evaluated the effect of plant growth-promoting (PGP) strains of Streptomyces on yield, quality, and nitrate content of fruits, plant-microbe responses, and antifungal effect against blight disease caused by fungus pathogen Alternaria solani on tomato fruits in commercial greenhouse conditions. Greenhouse trials were done with four treatments including strains Y28, IC10, IT25, and commercial bio-fertilizer (Barvar NPK®) on tomato plants. In PGP treatments, the number of infected fruits significantly reduced (60%) compared to Barvar and control. Strain Y28 improved the quality of tomatoes more than other treatments. All three PGP treatments contained a higher level of total sugar concentration and antioxidant enzyme activities than Barvar and control. In contrast, PGP strains, especially Y28, significantly reduced nitrate accumulation (25%) compared to Barvar and control tomatoes. Streptomyces treatments induced more than a 20-fold increase in UDP and WRKY70 transcription factor gene expression relative to the control (P < 0.01). Based on the results, microbe-dependent plant defense induced by these strains is positively correlated to WRKY70 expression and nitrate reduction in commercial greenhouse conditions. These findings suggest that the commercial application of specific strains not only can illustrate an eco-friendly solution to induce resistance against fungal pathogens but also improve the quality properties of food plants with lower nitrate content.

Induction of apoptosis and cell cycle arrest in colorectal cancer cells by novel anticancer metabolites of Streptomyces sp. 801.

BACKGROUND: Colorectal cancer is the third and most significant cause of death and fourth most common cancer in the world. Chemotherapy can be introduced in the cases of locally or distantly invasive colorectal cancer. In recent years Actinomycetes, especially the genus Streptomyces, contain numerous bioactive compounds, some of which are known as important anti-tumor chemotherapy drugs. In this research, we aimed to explore the anti-cancer mode of action of Streptomyces sp. 801 on colorectal cancer cells in vitro conditions. METHODS: Fermented supernatant of strain Streptomyces sp. 801 isolated from soil showed maximum growth inhibition on human colorectal cancer cells. The cytotoxic effects of various concentrations of EtOAc extract from bacterial culture supernatant on HT-29, HCT 116 and SW480 cancer cells were surveyed using the MTT assay. Moreover, flow cytometry assays and Bax, Bcl-2, Cyclin D1 and P21 gene expressions were carried out to assess the apoptotic and cell cycle effects. Also, the scratch assay was performed to measure migration. Finally, Ethyl acetate (EtOAc) extract was analyzed by LC-MS to identify anti-cancer compounds. RESULTS: The cell viability of all three cell lines were decreased in a dose-dependent manner. The successful induction of apoptosis and cell cycle arrest at IC50 values, were confirmed by flow cytometry as well as by the mRNA expression levels of the genes involved in these processes. Scratch assays indicated the inhibition of cell migration in the cancer cell lines treated by Streptomyces sp. 801. Nine anti-cancer compounds of Streptomyces sp. 801 were detected by liquid chromatography-mass spectrometry (LC-MS) analysis. CONCLUSIONS: These findings suggest that Streptomyces sp. 801 can be a source of promising anticancer metabolites.

Streptomyces strains modulate dynamics of soil bacterial communities and their efficacy in disease suppression caused by Phytophthora capsici.

The responses of rhizosphere bacterial communities of Streptomyces (SS14 and IT20 stains) treated-pepper plants following inoculation by Phytophthora capsici (PC) was investigated using Illumina MiSeq sequencing. Distinct modulation of the bacteriome composition was found for PC samples with the highest relative abundance (RA) of Chitinophaga (22 ± 0.03%). The RA of several bacterial operational taxonomic units (OTUs) was affected and caused changes in alpha and beta-diversity measures. In IT20, the RA of Cyanobacteria was enriched compared to SS14 (72%) and control samples (47%). Phylotypes belonging to Devosia, Promicromonospora, Kribbella, Microbacterium, Amylocolatopsis, and Pseudomonas genera in the rhizosphere were positively responding against the pathogen. Our findings show that the phosphate solubilizing strain IT20 has higher microbial community responders than the melanin-producing strain SS14. Also, positive interactions were identified by comparing bacterial community profiles between treatments that might allow designing synthetic bio-inoculants to solve agronomic problems in an eco-friendly way.

Neuroprotective effects of vitamin C and garlic on glycoconjugates changes of cerebellar cortex in lead-exposed rat offspring.

The deteriorating effects of Lead (Pb) on central nervous system (CNS) such as cerebellum has been demonstrated in previous studies. Glycoconjugates with the important role in CNS development may be affected by Pb-exposure. Utilization of antioxidant agents and herbal plants has attracted a great deal of attention on attenuating neurotoxicants-induced damage. Thus, in this study the neuroprotective effects of vitamin C and garlic on content of glycoconjugates of cerebellar cortex in Pb-exposed animals were investigated. Wistar pregnant rats were divided into: control (C), Pb-exposed (Pb) (1500 ppm lead acetate in drinking water), Pb plus vitamin C (Pb + Vit C) (500 mg/kg) intraperitoneally, Pb plus garlic (Pb + G) (1 mL /100 g body weight fresh garlic juice via gavage), Pb plus vitamin C and garlic (Pb + Vit C + G), and sham groups (Sh). Finally, levels of Pb in blood were measured in both rats and offspring on postnatal day 50 (PND50). Also, the cerebellums were removed for measuring Pb-levels and performing lectin histochemistry. Blood and cerebellar Pb-levels were increased in Pb-exposed group compared to control group (P < 0.001), whereas they were decreased significantly in Pb + Vit C, Pb + G, and Pb + Vit C + G groups (P < 0.01). By using MPA, UEA-1, and WGA lectin histochemistry, Pb-exposed group showed weak staining intensity compared to other groups. Besides, significant decrease was observed in the density of lectin-positive neurons of Pb-exposed group compared to the control group (P < 0.001). Moreover, strong staining intensity and high lectin-positive neurons were found in Pb + Vit C, Pb + G and Pb + Vit C + G groups than Pb-exposed group (P < 0.001). The present study revealed that Pb-exposure can result in alteration in the cerebellar glycoconjugates contents and co-administration of vitamin C and garlic could attenuate the adverse effects of Pb. The findings of this study revealed the ameliorating effects of vitamin C and garlic against Pb, suggesting the potential use of vitamin C and garlic as preventive agents in Pb poisoning.

Neurotrophic factors in Alzheimer's disease: pathogenesis and therapy.

Alzheimer's disease (AD) is a common neurodegenerative disease with a prevalence estimated to reach 115 million by 2050. It is characterized by abnormal extracellular accumulation of amyloid­beta (Aß) peptide and intracellular neurofibrillary tangles (NFTs) that result in neuro­inflammation, synaptic dysfunction, neurotransmitter imbalance, neuronal loss, and dendritic changes. A hypothesis of neurotrophic factor (NTF) involvement in neurodegenerative diseases and their potential as a therapeutic tool has emerged. There are wide information gaps on this topic. However, consistent with this hypothesis, AD may be caused by a deficiency in neurotrophin proteins or receptors expression. In AD brains, an increase in nerve growth factor and a decrease in brain-derived neurotrophic factor in the hippocampus and certain neocortical regions, and a decrease in TrkA in the cortex and nucleus basalis has been observed. Thus, comparative data relating to recent hypotheses addressing NTF content and receptors in experimental animals and human brains, along with their potential roles in the treat ment of AD, are discussed in this review.

Ectoine production in bioreactor by Halomonas elongata DSM2581: Using MWCNT and Fe-nanoparticle.

Halomonas elongate produces ectoine to protect itselt from environmental stresses. In this research, important factors in the production of ectoine were optimized using statistical methods to achieve the best production efficiency in bioreactor. Screening important variables (ectoine, hydroxyectoine, l-aspartic acid, and glutamate) on H. elongate growth showed that ectoine and l-aspartic acid directly affect ectoine production. Two nanostructures, multiwalled carbon nanotube (MWCNT) and iron oxide nanoparticle (Fe2 O3 NPs), were used to increase the availability of substrate for the microorganism. The results showed that Fe2 O3 nanoparticles and MWCNT could have a negative or positive effect on bacterial growth and ectoine production depending on the concentration of nanoparticles. At optimized conditions, the amounts of bacterial growth and ectoine production in fermenter were 10.4 g/L and 14.25 g/L, respectively. Therefore, it could be concluded that nanoparticles improve bacterial growth and ectoine production at optimized concentrations.

Testicular tuberculosis: An uncommon complication after treatment of urothelial carcinoma.

Tuberculous epididymo-orchitis is an uncommon complication after intravesical bacilli Calmette-Guerin therapy for nonmuscle invasive bladder cancer. Spread of granulomatous disease through the genitourinary tract specifically to the testes occurs in 0.4% of treated patients. The following case presents a 77-year-old man who underwent intravesical therapy after transurethral resection of bladder tumor and developed testicular discomfort and a palpable mass 2 years after initiation of therapy. After wide range of serum and urine analyses, repeated testicular ultrasonography, and an unsuccessful course of antibiotics, the patient elected to undergo orchiectomy and was confirmed to have tuberculous epididymo-orchitis. Diagnosis based on imaging and laboratory serum and urine analysis may be elusive and therefore review of this entity and associated sonographic findings is discussed.

Corrigendum to "Plant growth promoting Streptomyces strains are selectively interacting with the wheat cultivars especially in saline conditions" [Heliyon 6 (2) (2020) e03445].

[This corrects the article DOI: 10.1016/j.heliyon.2020.e03445.].

Tissue-specific synergistic bio-priming of pepper by two Streptomyces species against Phytophthora capsici.

Among several studied strains, Streptomyces rochei IT20 and S. vinaceusdrappus SS14 showed a high level of inhibitory effect against Phytophthora capsici, the causal agent of pepper blight. The effect of two mentioned superior antagonists, as single or combination treatments, on suppression of stem and fruit blight diseases and reproductive growth promotion was investigated in pepper. To explore the induced plant defense reactions, ROS generation and transcriptional changes of selected genes in leaf and fruit tissues of the plant were evaluated. The plants exposed to the combination of two species responded differently in terms of H2O2 accumulation and expression ratio of GST gene compared to single treatments upon pathogen inoculation. Besides, the increment of shoot length, flowering, and fruit weight were observed in healthy plants compared to control. Likely, these changes depended on the coordinated relationships between PR1, ACCO genes and transcription factors WRKY40 enhanced after pathogen challenge. Our findings indicate that appropriate tissue of the host plant is required for inducing Streptomyces-based priming and relied on the up-regulation of SUS and differential regulation of ethylene-dependent genes.

Plant growth promoting Streptomyces strains are selectively interacting with the wheat cultivars especially in saline conditions.

Plant growth promoting (PGP) effect of Streptomyces on wheat growth in different conditions has been mostly reported although mechanisms which caused wheat cultivars differently response to a PGP Streptomyces has been less studied. In this study, the effect of two Streptomyces strains, previously reported as PGPR, on the growth of four salt-sensitive commercial wheat cultivars under normal and saline conditions was investigated. Strain C-2012 differently affected the growth of the cultivars in the normal and stress conditions. Cultivars Gonbad with the highest (63%) and Zarin without increased dry biomass upon C-2012 treatments were selected for further study. Salinity significantly decreased seedling fresh and dry weight, K+ and chlorophyll content and glutathione S-transferase activity. Moreover, the stress increased proline and Na+ content and peroxidase (POX) and ascorbate peroxidase (APX) activity in both cultivars. Strain C-2012, generally, ameliorated the negative effect of the stress with increased chlorophyll and carotenoid and reduced Na+ content and APX and SOD activity in both cultivars, however, its effect on biomass was different. Increase in SOD, APX and POX activities in bacterial inoculated-Zarin, but not Gonbad, under normal conditions suggested that this cultivar may recognize strain C-2012 as a gentle stressor and not as a PGPR. These results showed that the responses of the wheat cultivars to a defined PGPR is different in the physiological, phenotypic and molecular level. Based on the results, the evaluation of the effect of a bio-fertilizer on each wheat cultivar is necessary prior to use in a commercial field.

Volumetric investigation of the hippocampus in rat offspring due to diabetes in pregnancy-A stereological study.

BACKGROUND: The brain development during the prenatal period is affected by various factors, including the mother's metabolic condition. It has been revealed that diabetes in pregnancy is associated with structural and functional alterations in offspring's hippocampus. Hippocampus, as a critical region with well-known roles in learning and memory consolidation, is vulnerable to changes in glucose level. This study was designed to investigate the effects of maternal diabetes during the pregnancy period and insulin therapy on the neuronal density and the volume of different subfields of the hippocampus in rat offspring at postnatal day 14 (P14). METHODS: Wistar female rats were randomly divided into diabetics (STZ-D), diabetes treated with insulin (STZ-INS) group, and controls (CON). The animals in all groups were mated by non-diabetic male rats. Two weeks after birth, male pups from each group were sacrificed. The Cavalieri method was carried out to estimate the total volume, and the numerical density of the neurons in the hippocampus and its sub regions was measured by the optical dissector technique. RESULTS: Bilateral hippocampal volume decreased in the diabetic group, mainly in the CA1, dentate gyrus (DG) and subiculum areas (P ≤ 0.05), when compared to control and insulin-treated diabetic animals. In all hippocampus sub-regions, maternal diabetes resulted in a significant decrease in the number of cells in comparison with two other groups (P ≤ 0.05 each). CONCLUSION: These data indicate that diabetes during pregnancy has a negative impact on the development of the hippocampus in the rats. These changes in the volume of hippocampal CA1, DG, and subiculum areas might be at the core of underlying neurocognitive and neurobehavioral impairments observed in the children of diabetic mothers.

Streptomyces Strains Induce Resistance to Fusarium oxysporum f. sp. lycopersici Race 3 in Tomato Through Different Molecular Mechanisms.

Plant growth promoting rhizobacteria (PGPR) are potential natural alternatives to chemical fungicides in greenhouse production via inducing plant immune system against biotic stresses. In this research, 126 Streptomyces isolates were recovered from rhizosphere soils of 13 different commercial vegetable greenhouses in Iran. Streptomyces isolates were screened for in vitro Plant growth promoting (PGP) traits and ability to antagonize Fusarium oxysporum f. sp. lycopersici race 3 (FOL), the causal agent of Fusarium wilt of tomato (FWT). Six isolates with the highest antagonistic activity and at least three PGP traits were selected and compared with chemical fungicide Carbendazim® in a greenhouse experiment. All bacterial treatments mitigated FWT disease symptoms like chlorosis, stunting and wilting at the same level or better than Carbendazim®. Strains IC10 and Y28 increased shoot length and shoot fresh and dry weight compared to not inoculated control plants. Phenotypic characterization and 16S rRNA gene sequencing showed, strains IC10 and Y28 were closely related to S. enissocaesilis and S. rochei, respectively. The ability of the superior biocontrol strains to induce antioxidant enzymes activity and systemic resistance (ISR) was investigated. Increased activity of catalase (CAT) in plant treated with both strains as well as an increase in peroxidase (POX) activity in plants treated with Y28 pointed to a strain specific-induced systemic resistance (ss-ISR) in tomato against FOL. The differential induced expression of WRKY70 and ERF1 (two transcription factors involved in plant defense) and LOX and TPX by the analyzed Streptomyces strains, especially after inoculation with FOL, suggests that ss-ISR is triggered at the molecular level.

The effects of maternal diabetes and insulin treatment on neurogenesis in the developing hippocampus of male rats.

Diabetes in pregnancy is associated with an increasing risk of congenital malformations and central nervous system disorders (CNS) especially hippocampal neuronal circuitry disruption as a discreet region involved in neurogenesis phenomenon. This study aimed to investigate the effect of maternal diabetes and insulin treatment on the expression and distribution pattern of NeuN and DCX as two important markers of neurogenesis paradigm in developing rat hippocampus. All animals were randomly divided into three groups as follows: Control group, Diabetic (STZ-D), Diabetic treated with insulin (STZ-INS). Diabetes was induced in Wistar female rats by Sterptozotocin intraperitoneal injection (single does). Following confirmation of diabetes, animals were mated with non-diabetic males. Four to six units of protamine-Zinc insulin were delivered subcutaneously (SC) in insulin treated group. At the post-natal day 14 (P14), the brain of male offspring's were removed for further study. In fact Immunofluorescence staining and Real time - PCR assays are used for evaluation of neurogenesis phenomenon. Our results showed a significant higher level of hippocampal DCX expression and an increase in the mean number of DCX positive cells in the DG of diabetic group male offspring (P < 0.05). We also found an insignificant up-regulation in the expression of DCX and the mean number of positive cells in the insulin-treated diabetic group neonates as compared to control group (P > 0.05). Nevertheless the results of immunofluorescence staining for NeuN also indicated that the mean number of NeuN+ cells was significantly lower in dentate gyrus of diabetic group male offspring (P < 0.05). Besides, there were significant down- regulation in the hippocampal mRNA expression of NeuN in diabetic pups compare to control (P < 0.05 each). Our results revealed that diabetes during pregnancy has an adverse effect on the hippocampal neurogenesis in rat neonates. Furthermore, the control of glycemia by insulin is sufficient to prevent the alterations in expression of neurogenesis markers.

Correction to: Plant growth promoting activity of an auxin and siderophore producing isolate of Streptomyces under saline soil conditions.

Unfortunately, one of the author's name had been published incorrectly as "Peyman Abaszadeh Dahaji". The correct name is " Payman Abbaszadeh Dahaji.