Analysis of the Inhibitory Effect of hsa-miR-145-5p and hsa-miR-203a-5p on Imatinib-Resistant K562 Cells by GC/MS Metabolomics Method.

Imatinib (IM) resistance is considered to be a significant challenge in the management of chronic myeloid leukemia (CML). Previous studies have reported that hsa-miR-145-5p and hsa-miR-203a-5p can overcome IM resistance and hsa-miR-203a-5p can alter glutathione metabolism in IM-resistant cells. The purpose of this study was to examine whether hsa-miR-145-5p or hsa-miR-203a-5p counters IM resistance by targeting the overall metabolic profile of IM-resistant K562 cells. The metablic profiling of cell lysates obtained from IM-sensitive, IM-resistant, and miR-transfected IM-resistant K562 cells was carried out using the GC-MS technique. Overall, 75 major metabolites were detected, of which 32 were present in all samples. The pathway analysis of MetaboAnalyst 5.0 revealed that the majorly enriched pathways included glucose metabolism, fatty acid biosynthesis, lipogenesis, and nucleotide metabolism. Eleven of identified metabolites, l-glutamine, l-glutamic acid, l-lactic acid, phosphoric acid, 9,12-octadecadienoic acid, 9-octadecenoic acid, myristic acid, palmitic acid, cholesterol, and ß-alanine, appeared in enriched pathways. IM-resistant cells had comparatively higher concentrations of all of these metabolites. Notably, the introduction of hsa-miR-145-5p or hsa-miR-203a-5p into resistant cells resulted in a decrease in levels of these metabolites. The efficacy of miR-203a-5p was particularly remarkable in comparison with miR-145-5p, as evidenced by partial least-squares-discriminant analysis (PLS-DA), which showed a high level of similarity in metabolic profile between IM-sensitive K562 cells and IM-resistant cells transfected with hsa-miR-203a-5p. The results indicate that GC-MS-based metabolic profiling has the potential to distinguish between drug-resistant and -sensitive cells. This approach can also be used to routinely monitor therapeutic response in drug-resistant patients, thus, enabling personalized therapy.

MicroRNAs Involved in Nutritional Regulation During Plant-Microbe Symbiotic and Pathogenic Interactions with Rice as a Model.

Plants are constantly challenged with numerous adverse environmental conditions, including biotic and abiotic stresses. Coordinated regulation of plant responses requires crosstalk between regulatory pathways initiated by different external cues. Stress induced by excessiveness or deficiency of nutrients has been shown to positively or negatively interact with pathogen-induced immune responses. Also, colonization by arbuscular mycorrhizal (AM) fungi can improve plant nutrition, mainly phosphorus and resistance to pathogen infection. The proposed review addresses these issues about a new question that integrates adaptation to nutrient stress and disease resistance. The main goal of the current review is to provide insights into the interconnected regulation between nutrient signaling and immune signaling pathways in rice, focusing on phosphate, potassium and iron signaling. The underpinnings of plant/pathogen/AM fungus interaction concerning rice/M. oryzae/R. irregularis is highlighted. The role of microRNAs (miRNAs) involved in Pi (miR399, miR827) and Fe (miR7695) homeostasis in pathogenic/symbiotic interactions in rice is discussed. The intracellular dynamics of membrane proteins that function in nutrient transport transgenic rice lines expressing fluorescent protein fusion genes are outlined. Integrating functional genomic, nutritional and metal content, molecular and cell biology approaches to understand how disease resistance is regulated by nutrient status leading to novel concepts in fundamental processes underlying plant disease resistance will help to devise novel strategies for crop protection with less input of pesticides and fertilizers.

Adrenal adenoma associated with raised metanephrine level: A case report.

INTRODUCTION AND IMPORTANCE: Adrenal adenomas are benign neoplasms of the adrenal cortex which can be functional or non-functional. The functional tumors manifest as Cushing syndrome. Rarely, they are associated with high levels of metanephrine due to pheochromocytoma. Here, we report an extremely rare case of adrenal adenoma associated with raised metanephrine, but no histopathological evidence of medullary changes. CASE PRESENTATION: A 30-year female with diabetes, hypothyroidism, and hypertension was admitted for resistant hypertension, severe refractory hypokalemia, decreased sleep, fearfulness, anxiety, and palpitation. Facial puffiness, ecchymoses on upper and lower limbs, and bilateral pedal edema were present. Laboratory investigation revealed raised 24-hour-urine cortisol (52.5 µmol/dL) and metanephrine (56.00 µmol/24 h), very low ACTH (<5.0 pg/mL), and negative dexamethasone suppression test. MRI brain findings were normal. CT scan of the abdomen revealed a well-defined soft tissue lesion in the right adrenal gland and diffuse osteopenia vertebrae. Hence, diagnosis of Cushing Syndrome secondary to adrenal adenoma, and pheochromocytoma was made, and laparoscopic adrenalectomy was done under pheochromocytoma protocol. The patient improved dramatically, thereafter. Histopathological findings showed adrenocortical adenoma only; no evidence of pheochromocytoma was found. CLINICAL DISCUSSION: We came across many articles reporting adrenal adenoma associated with high levels of metanephrine due to co-existing pheochromocytomas either in the ipsilateral or contralateral adrenal gland. But, in our case, the adrenal adenoma was associated with high metanephrine levels with no histopathologic features of pheochromocytoma. CONCLUSION: Cushing syndrome due to Cortisol secreting Adrenal adenoma can be associated with biochemical and clinical features suggestive of Pheochromocytoma which could resolve after adrenalectomy.

Complete small intestinal obstruction due to band formation in low-grade appendiceal mucinous neoplasm: A rare case report and literature review.

INTRODUCTION: Intestinal obstruction due to external compression is a rare condition, and those caused by appendiceal neoplasms like low-grade appendiceal mucinous neoplasms are extremely rare. PRESENTATION OF CASE: A 67-year-old post-menopausal female, with no history of prior abdominal surgery, presented to the emergency room with constipation for 2 days, and peri-umbilical pain and non-projectile bilious vomiting for 19 h. There was mild abdominal distension and diffuse tenderness. After ultrasonography and Computed Tomography (CT) of the abdomen and pelvis, a provisional diagnosis of complete small bowel obstruction secondary to perforated mucinous neoplasm of the appendix was made. An emergency exploratory laparotomy was performed followed by ileo-cecal resection and ileocolic anastomosis. Intraoperative findings revealed bands extending from the appendix to the ileum forming a closed loop with gangrenous spots in the distal ileum and ascitic fluid with mucin. Later, histopathological examination findings were consistent with low-grade appendiceal mucinous neoplasm (LAMN). DISCUSSION: We reviewed three cases of intestinal obstruction, caused by compression from a LAMN, including the presented case. Preoperative diagnosis of the cause of small intestinal obstruction on CT is sometimes challenging. Hence, in patients with intestinal obstruction with a transition point in the right lower quadrant of the abdomen on CT, a high index of suspicion for an appendiceal etiology is required. CONCLUSION: LAMN with an associated band should be considered as one of the differential diagnoses in the patient presenting with symptoms of complete small intestinal obstruction without prior abdominal surgery.

Hepatogastric fistula: a complication of pyogenic liver abscess in a patient with the Brugarda syndrome - A rare case report.

Hepatogastric fistula following pyogenic liver abscess (PLA) is a rare and fatal complication, and only a handful of cases have been reported without co-existing comorbidities of Brugarda syndrome. Case presentation: A 22-year-old male presented to the emergency room with a known case of Brugarda pattern ECG with chief complaints of on-and-off abdominal pain and fever for 2 weeks and shortness of breath for one day. On evaluation, echocardiography showed a clot in the inferior vena cava (IVC) and right atrium (RA), and on computed tomography scan of the abdomen revealed a liver abscess with transmural gastric perforation. During, an exploratory laparotomy where a fistula joining the left lobe of the liver and stomach was detected, and an emergency excision was done. The patient was shifted to the ICU and later developed septic shock, which was managed medically. Clinical discussion: Usually, thrombosis of the portal vein and the hepatic vein is a very common complication of a PLA but vascular complications like IVC, RA thrombosis, and hepatogastric fistula have been reported rarely. Our case is peculiar hepatogastric fistulization along with IVC/RA clots in a patient with Brugarda pattern ECG. The typical clinical manifestation of a patient with hepatogenic fistula is absent in our patient and presented with an on-off type of fever, epigastric pain, and shortness of breath and was managed surgically. Conclusion: Hepatogasric fistula, thrombosis of the IVC, and RA are a rare complications of PLA. The patient with Brugarda syndrome is at high risk as its clinical manifestation gets exaggerated during sepsis.

Antileukemic Activity of hsa-miR-203a-5p by Limiting Glutathione Metabolism in Imatinib-Resistant K562 Cells.

Imatinib has been the first and most successful tyrosine kinase inhibitor (TKI) for chronic myeloid leukemia (CML), but many patients develop resistance to it after a satisfactory response. Glutathione (GSH) metabolism is thought to be one of the factors causing the emergence of imatinib resistance. Since hsa-miR-203a-5p was found to downregulate Bcr-Abl1 oncogene and also a link between this oncogene and GSH metabolism is reported, the present study aimed to investigate whether hsa-miR-203a-5p could overcome imatinib resistance by targeting GSH metabolism in imatinib-resistant CML cells. After the development of imatinib-resistant K562 (IR-K562) cells by gradually exposing K562 (C) cells to increasing doses of imatinib, resistant cells were transfected with hsa-miR-203a-5p (R+203). Thereafter, cell lysates from various K562 cell sets (imatinib-sensitive, imatinib-resistant, and miR-transfected imatinib-resistant K562 cells) were used for GC-MS-based metabolic profiling. L-alanine, 5-oxoproline (also known as pyroglutamic acid), L-glutamic acid, glycine, and phosphoric acid (Pi)-five metabolites from our data, matched with the enumerated 28 metabolites of the MetaboAnalyst 5.0 for the GSH metabolism. All of these metabolites were present in higher concentrations in IR-K562 cells, but intriguingly, they were all reduced in R+203 and equated to imatinib-sensitive K562 cells (C). Concludingly, the identified metabolites associated with GSH metabolism could be used as diagnostic markers.

Wheat grain proteomic and protein-metabolite interactions analyses provide insights into plant growth promoting bacteria-arbuscular mycorrhizal fungi-wheat interactions.

KEY MESSAGE: Proteomic, protein-protein and protein-metabolite interaction analyses in wheat inoculated with PGPB and AMF identified key proteins and metabolites that may have a role in enhancing yield and biofortification. Plant growth-promoting bacteria (PGPB) and arbuscular mycorrhizal fungi (AMF) have an impact on grain yield and nutrition. This dynamic yet complex interaction implies a broad reprogramming of the plant's metabolic and proteomic activities. However, little information is available regarding the role of native PGPB and AMF and how they affect the plant proteome, especially under field conditions. Here, proteomic, protein-protein and protein-metabolite interaction studies in wheat triggered by PGPB, Bacillus subtilis CP4 either alone or together with AMF under field conditions was carried out. The dual inoculation with native PGPB (CP4) and AMF promoted the differential abundance of many proteins, such as histones, glutenin, avenin and ATP synthase compared to the control and single inoculation. Interaction study of these differentially expressed proteins using STRING revealed that they interact with other proteins involved in seed development and abiotic stress tolerance. Furthermore, these interacting proteins are involved in carbon fixation, sugar metabolism and biosynthesis of amino acids. Molecular docking predicted that wheat seed storage proteins, avenin and glutenin interact with secondary metabolites, such as trehalose, and sugars, such as xylitol. Mapping of differentially expressed proteins to KEGG pathways showed their involvement in sugar metabolism, biosynthesis of secondary metabolites and modulation of histones. These proteins and metabolites can serve as markers for improving wheat-PGPB-AMF interactions leading to higher yield and biofortification.

Bacillus subtilis impact on plant growth, soil health and environment: Dr. Jekyll and Mr. Hyde.

The increased dependence of farmers on chemical fertilizers poses a risk to soil fertility and ecosystem stability. Plant growth-promoting rhizobacteria (PGPR) are at the forefront of sustainable agriculture, providing multiple benefits for the enhancement of crop production and soil health. Bacillus subtilis is a common PGPR in soil that plays a key role in conferring biotic and abiotic stress tolerance to plants by induced systemic resistance (ISR), biofilm formation and lipopeptide production. As a part of bioremediating technologies, Bacillus spp. can purify metal contaminated soil. It acts as a potent denitrifying agent in agroecosystems while improving the carbon sequestration process when applied in a regulated concentration. Although it harbours several antibiotic resistance genes (ARGs), it can reduce the horizontal transfer of ARGs during manure composting by modifying the genetic makeup of existing microbiota. In some instances, it affects the beneficial microbes of the rhizosphere. External inoculation of B. subtilis has both positive and negative impacts on the endophytic and semi-synthetic microbial community. Soil texture, type, pH and bacterial concentration play a crucial role in the regulation of all these processes. Soil amendments and microbial consortia of Bacillus produced by microbial engineering could be used to lessen the negative effect on soil microbial diversity. The complex plant-microbe interactions could be decoded using transcriptomics, proteomics, metabolomics and epigenomics strategies which would be beneficial for both crop productivity and the well-being of soil microbiota. Bacillus subtilis has more positive attributes similar to the character of Dr. Jekyll and some negative attributes on plant growth, soil health and the environment akin to the character of Mr. Hyde.

Bacillus sp. and arbuscular mycorrhizal fungi consortia enhance wheat nutrient and yield in the second-year field trial: Superior performance in comparison with chemical fertilizers.

AIMS: The aim of the study is to analyse the effect of microbial consortia for wheat biofortification, growth, yield and soil fertility as part of a 2-year field study and compare it with the use of chemical fertilizers. METHODS AND RESULTS: A field trial (second year) was conducted with various combinations of plant growth-promoting bacteria (PGPB) and arbuscular mycorrhizal fungi (AMF) treatments, ranging from a single inoculant to multiple combinations. The microbial consortia used were Bacillus sp. and AMF based on first-year field trial results. The consortia based on native (CP4) and non-native (AHP3) PGPB (Bacillus sp.) and AMF performed better in terms of nutrients content in wheat grain tissue and yield-related traits compared with chemical fertilizer treated and untreated control. Dual treatment of PGPB (CP4+AHP3) combined with AMF resulted in a significant increase in antioxidants. The spatial colonization of AMF in roots indicated that both the isolates CP4 and AHP3 were able to enhance the AMF colonization in root tissue. Furthermore, soil enzymes' activities were higher with the PGPB and AMF combination giving the best results. A positive correlation was recorded between plant growth, grain yield and soil physicochemical parameters. CONCLUSIONS: Our findings confirm that the combined treatment of CP4 and AHP3 and AMF functions as an effective microbial consortium with excellent application prospects for wheat biofortification, grain yield and soil fertility compared with chemical fertilizers. SIGNIFICANCE AND IMPACT OF STUDY: The extensive application of chemical fertilizers on low-yielding field sites is a severe concern for cereal crops, especially wheat in the Asian continent. This study serves as a primer for implementing site-specific sustainable agricultural-management practices using a green technology leading to significant gains in agriculture.

Co-occurrence and patterns of phosphate solubilizing, salt and metal tolerant and antibiotic-resistant bacteria in diverse soils.

Soil is a treasure chest for beneficial bacteria with applications in diverse fields, which include agriculture, rhizoremediation, and medicine. Metagenomic analysis of four soil samples identified Proteobacteria as the dominant phylum (32-52%) followed by the phylum Acidobacteria (11-21% in three out of four soils). Bacteria that were prevalent at the highest level belong to the genus Kaistobacter (8-19%). PICRUSt analysis predicted KEGG functional pathways associated with the metagenomes of the four soils. The identified pathways could be attributed to metal tolerance, antibiotic resistance and plant growth promotion. The prevalence of phosphate solubilizing bacteria (PSB) was investigated in four soil samples, ranging from 26 to 59% of the total culturable bacteria. The abundance of salt-tolerant and metal-tolerant bacteria showed considerable variation ranging from 1 to 62% and 4-69%, respectively. In comparison, the soil with the maximum prevalence of temperature-tolerant and antibiotic-resistant bacteria was close 30%. In this study, the common pattern observed was that PSB were the most abundant in all types of soils compared to other traits. Conversely, most of the isolates, which are salt-tolerant, copper-tolerant, and ampicillin-resistant, showed phosphate solubilization activity. The sequencing of the partial 16S-rRNA gene revealed that PSB belonged to Bacillus genera. SUPPLEMENTARY INFORMATION: The online version contains supplementary material available at 10.1007/s13205-021-02904-7.

Assessment of Air Pollution Tolerance Index (APTI) and Anticipated Performance Index (API) of Roadside Plants for the Development of Greenbelt in Urban Area of Bathinda City, Punjab, India.

The aim of the present study is to biomonitor the tolerance level of roadside plant species towards mitigation of air pollution for the development of greenbelt. The leaves of four plant species (Azadirachta indica, Ficus benghalensis, Melia azedarach and Polyalthia longifolia) were collected from 41 sampling sites of Bathinda city during summer and winter seasons. Seasonal variation in APTI and API was determined in selected plants. Highest APTI (13.54) was reported in F. benghalensis during summer and lowest APTI (11.69) in A. indica during winter season. The APTI and API value revealed that F. benghalensis was the best performer, A. indica and M. azedarach were excellent performers and P. longifolia a good performer in greenbelt development to mitigate the air pollution. Pearson's correlation analysis illustrated significant correlation between APTI, ascorbic acid, total chlorophyll, relative water and pH, respectively.

Bacteria from native soil in combination with arbuscular mycorrhizal fungi augment wheat yield and biofortification.

Plant growth promoting bacteria (PGPB) have been used to enhance crop productivity. The effect of native PGPB and arbuscular mycorrhizal (AM) fungi in combination on wheat yield, biofortification and soil enzymatic activity is a relatively unexplored area. Twenty seven bacterial isolates from three different soils were characterized for their plant growth promoting traits. A total of three native and five non-native bacteria were used with and without arbuscular mycorrhizal (AM) fungi in an open greenhouse pot experiment with two wheat varieties to evaluate their effect on wheat yield, nutrient uptake, and soil health parameters. Wheat plants subjected to native PGPB (CP4) (Bacillus subtilis) and AM fungi treatment gave the best results with reference to macronutrient (nitrogen and phosphorus), micronutrient (iron and zinc) content in wheat grains and yield-related parameters, including thousand grain weight, number of grains per spike and total tillers per plant in both wheat cultivars. Treatment with CP4 and CP4 plus AM fungi enhanced total chlorophyll in wheat leaves indicating higher photosynthetic activity. Significant improvement in soil health-related parameters, including soil organic matter and dehydrogenase activity, was observed. Significant correlation among grain yield-related parameters, nutrient enhancement, and soil health parameters was observed in PGPB and AM fungi treated plants, especially HD-3086. These results provide a roadmap for utilizing native PGPB and AM fungi for enhancing wheat production in Punjab state of India and exploring their utility in other parts of the country with different soil and environmental conditions.

Brown gold of marginal soil: Plant growth promoting bacteria to overcome plant abiotic stress for agriculture, biofuels and carbon sequestration.

Marginal land is defined as land with poor soil characteristics and low crop productivity with no potential for profit. Poor soil quality due to the presence of xenobiotics or climate change is of great concern. Sustainable food production with increasing population is a challenge which becomes more difficult due to poor soil quality. Marginal soil can be made productive with the use of Plant Growth Promoting Bacteria (PGPB). This review outlines how PGPB can be used to improve marginal soil quality and its implications on agriculture, rhizoremediation, abiotic stress (drought, salinity and heavy metals) tolerance, carbon sequestration and production of biofuels. The feasibility of the idea is supported by several studies which showed maximal increase in the growth of plants inoculated with PGPB than to uninoculated plants grown in marginal soil when compared to the growth of plants inoculated with PGPB in healthy soil. The combination of PGPB and plants grown in marginal soil will serve as a green technology leading to the next green revolution, reduction in soil pollution and fossil fuel use, neutralizing abiotic stress and climate change effects.

Nutrient enhancement of chickpea grown with plant growth promoting bacteria in local soil of Bathinda, Northwestern India.

Plant growth promoting bacteria (PGPB) enhance crop productivity as part of green technology to reduce the use of chemical fertilizers. They also have the capability to enhance macro- and micronutrient content of plants. In the present study, PGPB isolates belonging to Pseudomonas citronellis (PC), Pseudomonas sp. RA6, Serratia sp. S2, Serratia marcescens CDP13, and Frateuria aurantia (Symbion-K) were tested on two chickpea varieties, PBG1 and PBG5 grown for 30 days in local soil from Bathinda region in Northwestern India. PC and CDP13 were found to be better chickpea growth stimulators compared to the commercial Symbion-K based on shoot length and biomass. Most PGPB enhanced macro- and micronutrients in shoots to varying degrees compared to the control. PBG5 gave better response compared to PBG1 with reference to plant growth attributes and enhancement of the macronutrients, calcium, nitrogen and phosphorus and micronutrients, boron, copper, iron, and zinc. PBG5 is a high yielding variety with better resistance compared to PBG1. Overall, PGPB isolated from the local soil and PGPB from other parts of India were shown to be useful for enhancement of nutrient content and plant growth.