Differential responses of chili varieties grown under cadmium stress.

Heavy metal cadmium (Cd) naturally occurs in soil and is a hazardous trace contaminant for humans, animals, and plants. The main sources of Cd pollution in soil include overuse of phosphatic fertilizers, manure, sewage sludge, and aerial deposition. That's why an experiment was conducted to analyze the effect of Cd toxicity in Capsicum annuum L. by selecting its seven varieties: Hybrid, Desi, Sathra, G-916, BR-763, BG-912, and F1-9226. Cadmium was spiked in soil with four levels, i.e., (0, 3, 4, and 5 mg Cd kg- 1 of soil) for a week for homogeneous dispersion of heavy metal. Chili seeds were sown in compost-filled loamy soil, and 25-day-old seedlings were transplanted into Cd-spiked soil. Cadmium increasing concentration in soil decreased chili growth characteristics, total soluble sugars, total proteins, and amino acids. On the other hand, the activities of antioxidant enzymes were increased with the increasing concentration of Cd in almost all the varieties. Treatment 5 mg Cd/kg application caused - 197.39%, -138.78%, -60.77%, -17.84%, -16.34%, -11.82% and - 10.37% decrease of carotenoids level in chili V2 (Desi) followed by V4 (G-916), V1 (Hy7brid), V7 (F1-9226), V6 (BG-912), V5 (BR-763) and V3 (Sathra) as compared to their controls. The maximum flavonoids among varieties were in V5 (BR-763), followed by V6 (BG-912), V7 (F1-9226), V3 (Sathra) and V1 (Hybrid). Flavonoids content was decreased with - 37.63% (Sathra), -34.78% (Hybrid), -33.85% (G-916), -31.96% (F1-9226), -31.44% (Desi), -30.58% (BR-763), -22.88% (BG-912) as compared to their control at 5 mg Cd/kg soil stress. The maximum decrease in POD, SOD, and CAT was - 31.81%, -25.98%, -16.39% in chili variety V7 (F1-9226) at 5 mg Cd/kg stress compared to its control. At the same time, maximum APX content decrease was - 82.91%, followed by -80.16%, -65.19%, -40.31%, -30.14%, -10.34% and - 6.45% in V4 (G-916), V2 (Desi), V3 (Sathra), V6 (BG-912), V1 (Hybrid), V7 (F1-9226) and V5 (BR-763) at 5 mg Cd/kg treatment as compared to control chili plants. The highest CAT was found in 5 chili varieties except Desi and G-916. Desi and G-916 varieties. V5 (BR-763) and V6 (BG-912) were susceptible, while V1 (Hybrid), V3 (Sathra), and V7 (F1-9226) were with intermediate growth attributes against Cd stress. Our results suggest that Desi and G-916 chili varieties are Cd tolerant and can be grown on a large scale to mitigate Cd stress naturally.

Assessing p-tolyloxy-1,3,4-oxadiazole acetamides as lipoxygenase inhibitors assisted by in vitro and in silico studies.

The underlying correlation between the inflammation, innate immunity and cancer is extensively familiar and linked through a process mediated by three enzymes; cyclooxygenase (COX), lipoxygenase (LOX) and cytochrome P450 (CYP450). The ever increase in the reported side effects of the antiinflammatory drugs against the targeted enzymes and the resistance developed afterwards compels the researchers to synthesize new effective molecules with safer profile. On the basis of these facts, our ongoing research on 1,3,4-oxadiazole derivatives deals with the synthesis of a new series of N-alkyl/aralky/aryl derivatives of 5-((p-tolyloxymethyl)-4H-1,3,4-oxadiazole-2-ylthio)acetamide (6a-o) which were developed by the sequential conversion of p-tolyloxyacetic acid (a) into ester (1) hydrazide (2) and 5-(p-tolyloxymethyl)-4H-1,3,4-oxadiazole-2-thiol (3). The designed compounds (6a-o) were acquired by the reaction of 1,3,4-oxadiazole (3) with numerous electrophiles (5a-o) in KOH. The synthesized analogues (6a-o) were characterized by FTIR, 1H-, 13C NMR spectroscopy, EI-MS and HR-EI-MS spectrometry, and were further assessed for their inhibitory potential against the soybean 15-LOX enzyme. The results showed excellent inhibitory potential of the compounds against the said enzyme, specifically 6o, 6b, 6n and 6e with inhibitory values (IC50 ± SEM) of 21.5 ± 0.76, 24.3 ± 0.45, 29.1 ± 0.65 and 31.3 ± 0.78 µM, respectively. These compounds displayed < 55 % blood mononuclear cells (MNCs) cellular viability as measured by MTT assay at 0.25 mM concentration. Other compounds demonstrated moderate inhibitory activities with IC50 values in the range of 33.2 ± 0.78 to 96.3 ± 0.73 µM and exhibited little cellular viability against MNCs except 6i, 6j, 6 m and 6 k that showed 61-79 % cellular viability. It was observed that most of the compounds (6o, 6b, 6n, 6e) were found more toxic towards MNCs at studied concentration of 0.25 mM. SAR studies revealed that the positions and nature of substituents accompanying phenyl ring have great influence on 15-LOX inhibitory activity. In the most active compound 6o, the amino acids Asp768 and Val126 were involved in hydrogen bonding, Thr529 was linked with π-anion interaction and π-sulphur interaction was displayed with Tyr525 and two π-alkyl interactions were formed with the benzene ring and amino acid residues Pro530 and Arg533. The in silico pharmacokinetics profiles and density functional theory calculations of the compounds further supported the in vitro findings. Further work on the synthesis of more oxadiazole derivatives is in progress in search for potential 'leads' for the drug discovery as LOX inhibitors.

Molecular and functional analysis of naphthalene-degrading bacteria isolated from the effluents of indigenous tanneries.

During present study, four naphthalene- metabolizing bacteria were isolated from tanneries effluents through enrichment on naphthalene as sole carbon source in minimal salt medium. The bacteria were analyzed to document growth pattern, naphthalene removal efficiency, biochemical and molecular characteristics, antibiotic sensitivity, and metabolic profile. The 16S ribosomal RNA gene sequences were compared through BLAST (basic local alignment search tool) similarity search tool and three isolates were found homologous to Brevibacillus agri strain NBRC 15538 and one similar to Burkholderia lata strain 383. The naphthalene removal efficiencies ranged from 1.16 ± 0.056 mg/h (IUBN1) to 1.379 ± 0.021 mg/h (IUBN26). All isolates were positive for p-nitrophenyl phosphate (PO4 ), esculin, and inulin fermentation tests. Majority were positive for glucosaminidase (IUBN3, 17, and 26) and a few for mannitol and sorbitol fermentation (IUBN1). Identification of metabolites through gas chromatography-mass spectrometry and liquid chromatography-mass spectrometry analysis allowed tracing pathways associated with naphthalene degradation. Intermediates such as cis-dihydrodiolnaphthalene, 2-hydroxychromene-2-carboxylate, 6-hydroxyhexanoic acid, acetyl-CoA confirmed that the present study bacteria can metabolize naphthalene through a pathway which differs from the pathways reported in earlier known bacteria. Due to fast growth rates, high naphthalene removal potentials, and multiple degradation pathways, these bacteria can be exploited for bioremediation of naphthalene.

Toluene degradation via a unique metabolic route in indigenous bacterial species.

Tanneries are the primary source of toluene pollution in the environment and toluene due to its hazardous effects has been categorized as persistent organic pollutant. Present study was initiated to trace out metabolic fingerprints of three toluene-degrading bacteria isolated from tannery effluents of Southern Punjab. Using selective enrichment and serial dilution methods followed by biochemical, molecular and antibiotic resistance analysis, isolated bacteria were subjected to metabolomics analysis. GC-MS/LC-MS analysis of bacterial metabolites helped to identify toluene transformation products and underlying pathways. Three toluene-metabolizing bacteria identified as Bacillus paralicheniformis strain KJ-16 (IUBT4 and IUBT24) and Brevibacillus agri strain NBRC 15538 (IUBT19) were found tolerant to toluene and capable of degrading toluene. Toluene-degrading potential of these isolates was detected to be IUBT4 (10.35 ± 0.084 mg/h), IUBT19 (14.07 ± 3.14 mg/h) and IUBT24 (11.1 ± 0.282 mg/h). Results of GC-MS analysis revealed that biotransformation of toluene is accomplished not only through known metabolic routes such as toluene 3-monooxygenase (T3MO), toluene 2-monooxygenase (T2MO), toluene 4-monooxygenase (T4MO), toluene methyl monooxygenase (TOL), toluene dioxygenase (Tod), meta- and ortho-ring fission pathways. But additionally, confirmed existence of a unique metabolic pathway that involved conversion of toluene into intermediates such as cyclohexene, cyclohexane, cyclohexanone and cyclohexanol. LC-MS analysis indicated the presence of fatty acid amides, stigmine, emmotin A and 2, 2-dinitropropanol in supernatants of bacterial cultures. As the isolated bacteria transformed toluene into relatively less toxic molecules and thus can be preferably exploited for the eco-friendly remediation of toluene.

Isolation and Characteristics of Biotechnologically Important Antagonistic Thermophilic Bacteria from Rhizosphere of Haloxylon salicornicum.

Rhizobacteria are an active part of microbial population in the rhizosphere of plants. In this study, twenty rhizobacteria were isolated from the rhizosphere of a perennial grass, Haloxylon salicornicum, found in Cholistan desert, an arid landmass near Bahawalpur Pakistan, in one set of experimental conditions. Colony characteristics, biochemical and molecular analyses of these isolates were performed. All isolates were bacilli, gram positive with off-white colonies and exhibited typical bacilli colony morphology. None of the isolates was gelatinase, urease, indole, H2S and catalase producer. Eleven isolates were amylase producers and 8 isolates were acid producers. All isolates fermented glucose, 3 fermented lactose and 19 fermented fructose. Molecular data revealed that out of twenty isolates, 14 isolates showed 91-99% identity with Brevibacillus borstelensis, 4 with Bacillus subtilis (97-98%) and 2 with Bacillus licheniformis (94-99%) through BLAST analysis. All identified bacterial isolates cladded with their respective groups in the phylogenetic tree. Many (11-15 out of 20) of the isolates were more effective in inhibiting growth of the tested bacterial strains as compared to the positive control (Ampicillin 50 µg/disc). We conclude that bacilli are the predominant form populating rhizosphere of this desert grass. Among the isolated bacteria Brevibacillus borstelensis, Bacillus subtilis and Bacillus licheniformis are the most predominant species.

Determination of nitrate in biological fluids by HPLC.

Nitrate is the stable product of nitric oxide, which is physiologically active radical, an immunomodulator and a neuromodulator; its quantification in biological fluids is important to study the physiological and biochemical nature. Therefore, the purpose of this study was to quantify nitrate in different biological fluids like serum, cerebrospinal fluid (CSF) and ascetic fluid (ASF) using HPLC technique. A new HPLC method for the estimation of nitrate in serum, CSF and ASF was developed using the mobile phase of 1.0mM each of Na2CO3 and NaHCO3 (1:1, v/v, pH 5 with H3PO4) at a flow rate of 1.0mLmin-1. Eluate was detected at 220nm with the retention time of nitrate 2.55 min. The LOD and LOQ values of nitrate were 0.03µgmL-1 and 0.098µgmL-1, respectively. Nitrate was eluted through SAX Hypersil column of 150 × 4.6mm, id, 5µm particle size. Run time was 10min. The method was validated according to the FDA guidelines and was found linear in the range of 0.39 to 50µgmL-1 and CV was <3%, within limits of FDA guidelines. The method was used successfully for the estimation of nitrate in biological fluids like serum, CSF and ASF of 20 patients each. This is an alternate and reproducible method for the detection of nitrates in biological fluids.

An Insight into the Role of E2F1 in Breast Cancer Progression, Drug Resistance, and Metastasis.

AIMS: This study aimed to investigate the role of E2F1 in breast cancer biology. BACKGROUND: Expression of E2F1, a transcription factor of many oncogenes and tumor suppressor genes, is lowered in several malignancies, including breast carcinoma. OBJECTIVES: In the present study, we analyzed the status of E2F1 expression in association with diverse attributes of breast malignancy and its impact on cancer progression. METHODS: For this purpose, we used various freely available online applications for gene enrichment, expression, and methylation analysis to extract mutation-based E2F1 map, to measure E2F1 drug sensitivity, and to determine E2F1 association with DNA damage response proteins. RESULTS: Results revealed tissue-specific regulatory behavior of E2F1. Moreover, the key role of E2F1 in the promotion of metastasis, stem cell-mediated carcinogenesis, estrogen-mediated cell proliferation, and cellular defense system, has therefore highlighted it as a metaplastic marker and hot member of key resistome pathways. CONCLUSION: The information thus generated can be employed for future implications in devising rational therapeutic strategies. Moreover, this study has provided a more detailed insight into the diagnostic and prognostic potential of E2F1.

Analysis of antibacterial and cytotoxic potential of medicinal plants from Cholistan desert, Pakistan.

This study aimed to investigate the antibacterial and cytotoxic activity of 03 medicinal plants, Calligonum polygonides, Farsetia hamiltonii, and Pulcaria crispa, from Cholistan desert, Pakistan. The active constituents of plants species were extracted in 05 different solvents and the extracts were tested against various bacterial strains and brine shrimps. Although all Calligonum polygonides's extracts except chloroform were active against Staphylococcus aureus the most active was the acetone extract (21 ± 0.00 mm at 200 µg/disc) and activity was better than Caricef (p-value 0.03). While its water extract was more potent (18 ± 1.45 mm at 200 µg/disc) than Augmentin and Caricef (p-value < 0.005). The methanol extract's activity (15 ± 0.39 mm in 200 µg/disc) was comparable to Fucidin against Proteus vulgaris (p-value > 0.99) and activity of diethyl ether extract against Escherichia coli (10 ± 1.16 mm in 200 µg/disc) was same as of Urixin (p-value 0.91). Farsetia hamiltonii's acetone extract against Pseudomonas aeruginosa (10 ± 0.15 mm in 1 µg/disc) was more active than Augmentin Caricef and Cefotax (p-value < 0.02) and against Staphylococcus aureus (15 ± 1.15 mm in 200 µg/disc) activity was higher than Caricef (p-value 0.03). All Pulicaria crispa's extracts except water extract were found active against Staphylococcus aureus. However, the diethyl ether extract was most effective (25 + 0.00 mm at 150 µg /disc) and activity was more than Augmentin, Oxy-tetracycline, Fucidin, Urixin, Ceftriaxone (p-value < 0.05). Although all extracts were exhibited cytotoxic activity, the Calligonum polygonides's acetone extract (100%), Farsetia hamiltonii's diethyl ether extract (90%) and Pulicaria crispa's methanol extract (100%) were most active at 1000 µg/ml concentration. This study validated the medicinal significance of the studied plants and thus opens the way for their therapeutic applications.

Investigations of p-tolyloxy-1,3,4-oxadiazole propionamides as soybean 15-lipoxygenase inhibitors in comforting with in vitro and in silico studies.

Inflammatory disorders are the prime contributor to public health issue and the development of more effective and safer anti-inflammatory drugs in addition to other therapeutic alternatives to treat inflammatory illnesses, particularly chronic inflammatory diseases, is one of the foremost current issues. In this regard, our present work is concerned with the synthesis of a new series of N-alkyl/aralkyl/aryl derivatives (7a-o) of 5-((p-tolyloxymethyl)-4H-1,3,4-oxadiazole-2-ylthio)propionamide which was instigated by the successive conversions of p-tolyloxyacetic acid into ester, hydrazide and 5-(p-tolyloxymethyl)-4H-1,3,4-oxadiazole-2-thiol. The planned compounds (7a-o) were attained by the reaction of 5-(p-tolyloxymethyl)-4H-1,3,4-oxadiazole-2-thiol with variety of N-alkyl/aralkyl/aryl electrophiles in potassium hydroxide and were characterized by FTIR, 1H-, 13C-NMR spectroscopy, EI-MS and HR-EI-MS spectrometry and probed for their inhibiting potential against soybean 15-lipoxygenase (15-LOX) enzyme. The compounds 7a, 7n, 7 g, 7e, 7h, 7i, 7j and 7b promulgated the potent inhibiting potential with IC50 values 9.43 ± 0.45, 16.75 ± 0.49, 19.45 ± 0.37, 21.32 ± 0.46, 22.64 ± 0.56, 23.53 ± 0.62, 24.32 ± 0.45 and 29.15 ± 0.57 µM, respectively, while excellent to good inhibitory activities were shown by 7o, 7 m, 7k, 7f, 7c, 7 l and 7d with IC50 values in the range 30.29 ± 0.56 to 52.54 ± 0.64 µM. Compounds 7i-o maintained 91.12 ± 1.5 to 98.23 ± 1.2% blood mononuclear cells (MNCs) viability at 0.25 mM by MTT assay whilst compounds 7d-h observed 46.51 ± 1.3 to 57.12 ± 1.4% viability where as the most toxic compounds were 7b (12.51 ± 1.4%), 7a (28.12 ± 1.5%) and 7c (38.23 ± 1.5%) as compared with controls. Pharmacokinetic profiles predicted good oral bioavailability and drug-likeness properties of molecules as per rule of five. Molecular docking studies displayed hydrogen bonding between the compounds and the enzyme with Arg378 which was common in 7n, 7 g, 7h and baicalein. In 7a and quercetin, hydrogen bonding was established through Asn375; Tyr512 and Val589 were also involved in bonding with other analogues. RMSD (root mean square deviation) values exhibited good inhibitory profiles in the order quercetin (0.73 Å)<7 g (0.98 Å)

Screening of hepatitis B and C viral infection, recognition of risk factors, and immunization of patients against hepatitis B virus: a module developed for effective hepatitis control.

Introduction: The continually increasing incidence of hepatitis, a worldwide health issue, in Pakistan, has highlighted the need to investigate the epidemiology factors and implement preventive measures accordingly. The purpose of this study was to scrutinize the prevalent and significantly associated risk factors of hepatitis in students and employees, screening them for hepatitis B and C virus and vaccinating them against HBV to make IUB hepatitis free. Methodology: A total of 12,912 participants including students (n = 10,948) and employees (n = 1964) were screened for HBV and HCV via immunochromatographic test. Hepatitis- positive participants' blood samples were further tested and viral load was estimated by quantitative PCR. All the hepatitis-negative participants were vaccinated against HBV. The demographic and risk factors-related data were collected using the questionnaire. Statistical analysis (Chi-square test and bivariate regression analysis) was performed using SPSS software to explore any association between risk factors and hepatitis. Results: Results indicated that 662/12912 participants (students = 478/10,948, employees = 184/1,964) tested positive for hepatitis. Among them, HCV was observed to be more prevalent than HBV among the study participants, employees, and students, and viral count was low in both HBV and HCV-infected participants. However, men were more affected than women. The studied risk factors represented higher frequency among hepatitis-positive participants relative to the hepatitis-negative participants. The Chi-square test revealed that students' gender, history of hepatitis in the family and relatives, dental treatment, sharing cosmetics and shaving blades were significant (p > 0.005) risk factors of hepatitis while in the employees group surgery and age were significant. Moreover, the reused of syringes was found to be associated with hepatitis in both groups. The bivariate analysis helped to identify various new risk factors which were independently, either positively or negatively, associated with hepatitis. Discussion: Our study enabled us to recognize different risk factors of hepatitis among the target population. The information thus generated can be usefully applied in planning hepatitis awareness, targeted screening, and effective control programs for other target populations. In general, this module can be further utilized for any other disease.

Mutational analysis of hemoglobin genes and functional characterization of detected variants, through in-silico analysis, in Pakistani beta-thalassemia major patients.

Thalassemia is one of the most prevalent genetic disorders worldwide. The present study aimed to explore the mutational spectrum of all hemoglobin (HB) encoding genes and to identify the potentially damaging and pathogenic variants in the beta (ß)-thalassemia major patients and thalassemia minor carriers of Southern Punjab, Pakistan. A total of 49 ß-thalassemia major patients and 49 carrier samples were screened for the identification of HBA1, HBA2, HBB, HBD, HBE1, HBG1 and HBG2 variants by NGS. PCR was performed for the amplification of HB encoding genes and the amplified product of 13 patients and 7 carrier samples were processed for the Sanger sequencing. Various bioinformatics tools and databases were employed to reveal the functional impact and pathogenicity potential of the observed variants. Results depicted a total of 20 variants of HB-related genes by NGS and 5 by Sanger sequencing in thalassemia patients. While 20 variants by NGS and 3 by Sanger were detected in carriers. Few known genetic variants of HB-encoding genes are being reported for the first time in Pakistani thalassemia patients and carriers. However, two novel HBB variants c.375A>C (p.P125P) and c.*61T>G and a novel variant of HBE1 (c.37A>T (p.T13S)) were also documented. Pathogenicity analysis predicted the pathogenic potential of HBB variants (c.47G>A (p.W16*), c.27-28insG (p. S10fs), and c.92+5G>C) for ß thalassemia. The study of functional impact indicated that these HBB variants result in the premature termination of translation leading to the loss of functional ß-globin protein. It is therefore suggested that the pathogenic HBB variants, identified during present study, can be employed for the diagnosis, carrier screening, and planning therapy of thalassemia.

Exceptional behavior of breast cancer-associated type 1 gene in breast invasive carcinoma.

Background: Cellular expression level of Breast Cancer-Associated Type 1 (BRCA1) encoded protein is the sign of genome integrity, stability, and surveillance. BRCA1 after sensing DNA damage activates repairing system and if mutated leaves genomic lesions unrepaired and triggers transformation of normal breast cells into cancerous ones. Aims of study: We conducted in silico study to have a holistic view of BRCA1's correlation with multiple variables of breast invasive carcinoma. Materials and Methods: We used user-friendly online GeneCardsSuite pathway-level enrichment analysis, UALCAN portal differential expression analysis, cBioPortal cancer genome platform for mutatome map construction, and cancer cell lines encyclopedia genomics of drug sensitivity toolkit to understand correlation of BRCA1 expression with the effectiveness of anti-cancer drugs. Results: Contrary to general behavior of a tumor suppressor gene our study revealed BRCA1 overexpression under all circumstances. This novel finding needs to be explored further to understand functional impact of BRCA1 overexpression on the expression of many genes which are transcriptionally regulated by BRCA1 and promotion of tumriogenesis. Conclusion: Our study highlights the potential role of BRCA1-regulated genes in oncogenesis and recommends use of BRCA1-linked genes as future therapeutic targets for effective disease management.

Identification of Key Biomarkers for the Future Applications in Diagnostics and Targeted Therapy of Colorectal Cancer.

BACKGROUND: Colorectal cancer (CRC) is one of the most prevalent and deadliest malignancies in the world. INTRODUCTION: The microarray dataset GSE87211 has been re-analyzed in the present study through a multi-layered bioinformatics approach to identify the CRC associated hub genes. GSE87211 dataset was retrieved and GEO2R was implemented for the identification of differentially expressed genes (DEGs). STRING tool was used to construct the protein-protein interaction (PPI) network. Cytoscape was utilized to identify the top six hub genes. METHOD: KEGG analysis was performed using DAVID. Expression validation of the hub genes and exploration of the correlation between hub genes expression and various other parameters was done through UALCAN, GEPIA, GENT2, cBioportal, TIMER, RegNetwork, and CTD. The six identified hub genes (GAL, GALR1, SST, SSTR2, NPY, and NPY1R) were enriched in diverse cancer-driving pathways. GAL was significantly up-regulated while other 5 hub genes (GALR1, SST, SSTR2, NPY, and NPY1R) were significantly down-regulated in colon adenocarcinoma (COAD) patients relative to controls. RESULT: All these hub genes were hypermethylated and 5 out of 6 hub genes were found to be associated with the worst Overall survival (OS) of the COAD patients relative to control group. Furthermore, we explored some interesting associations between hub genes' expression and different other parameters including copy number variations (CNVs), CD+T immune cells infiltration, different other cancer states and crucial mutant genes across COAD samples. In addition, a few miRNAs (miR-27a-3p and miR-130a-3p) and drugs (triclosan, soman, reserpine, and isoproterenol, etc.) were found capable to alter the expression of hub genes and thus need to further be evaluated for their potential role in CRC treatment. CONCLUSION: In conclusion, the identified hub genes may provide new insight into the CRC biology and treatment.

Phylogenetic Illustration of Eisenia fetida Associated Vermi-bacteria Involved in Heavy Metals Remediation and Retaining Plant Growth Promoting Traits.

Heavy metals contamination in the soil is a major threat to wildlife, the environment, and human health. Microbial remediation is an emerging and promising technology to reduce heavy metals toxicity. Therefore, the present research aimed to isolate and to identify the heavy metals tolerated bacteria from the Eisenia fetida for the first time, and to screen the bacto-remediation capabilities and plant growth promoting traits of vermi-bacterial isolates. Vermi-bacteria was isolated from the gut of E. fetida, identified through staining, culturing, biochemical tests, and ribotyping. Plant growth-promoting traits were also evaluated. Phylogenetic results revealed that isolated Vermi-bacterial strains showed resemblance with Bacillus thuringiensis, Bacillus aryabhattai, Staphylococcus hominis, Bacillus toyonensis, Bacillus cabrialesii, Bacillus tequilensis, Bacillus mojavensis, Bacillus amyloliquefaciens, Bacillus toyonensis, Bacillus anthracis, and Bacillus paranthracis. All identified Vermi-bacterial species are Gram-positive (rod and cocci) in nature, not only indicated the efficient biosorption of lead, cadmium, and chromium but also produce all plant growth stimulating traits such as indole acetic acid (IAA), amylase, protease, lipase, hydrogen cyanide, ammonia, and siderophore production, and also act as a phosphate solubilizers. Bacillus anthracis showed significant production of siderophore (33.0±0.0 mm), phosphate solubilizing (33.0±0.0 mm), proteolytic (15.0±0.0 mm), and lipolytic activities (20.0±0.0 mm) compared to other vermi-bacterial isolates. Bioaccumulation factor results revealed that Bacillus anthracis showed more accumulation of Cd (12.00±0.01 ppm), Cr (5.38±0.01 ppm), and Pb (4.38±0.01 ppm). Therefore, the current findings showed that all identified vermi-bacteria could be used as potential bactoremediation agents in heavy metals polluted environments and could be used as microbial biofertilizers to enhance crop production in a polluted area.

Hypothyroidism correlates with dyslipidemia and protein contents in patients with various metabolic disorders.

OBJECTIVES: The present study aimed to investigate the status of abnormalities in thyroid-related hormones, lipid profile parameters and total proteins in the sera of patients suffering from various metabolic disorders. METHODS: To analyze the study parameters, enzyme-linked immunosorbent assays, Bradford assays and standard clinical kits and methods were applied. Data were analyzed through the appropriate statistical tests. RESULTS: In all subjects except those with hypotension, thyroid-stimulating hormone, total cholesterol and low-density lipoprotein were elevated, whereas triiodothyronine levels were downregulated. Thyroxin was downregulated in subjects with diabetes and symptomatic thyroiditis but upregulated in patients with hypertension. High-density lipoprotein was upregulated in men who were diabetic only, and total protein was downregulated in those with hypotension only. Hypothyroidism in patients with diabetes, symptomatic thyroiditis and hypertension was correlated with dyslipidemia. In subjects with hypertension, it was correlated with total protein. CONCLUSION: This study revealed a link between hypothyroidism, dyslipidemia and total protein in patients with various metabolic disorders.

Molecular characterization of plant growth-promoting vermi-bacteria associated with Eisenia fetida gastrointestinal tract.

Earthworms are highly productive invertebrates and play a vital role in organic farming and improving soil structure and function. The gastrointestinal tract of earthworms possessed agricultural important bacteria. So, the current research aimed was to examine, screen, and identify the plant growth promoting bacteria existing in the digestive tract of Eisenia fetida called plant growth promoting vermi-bacteria. The plant growth promoting traits such as siderophore, phytohormone, and hydrolytic enzymes production, and phosphate solubiliation were assessed. Eleven vermi-bacteria i.e. Bacillus mycoides, B. aryabhattai, B. megaterium, Staphylococcus hominis, B. subtilis, B. spizizenii, B. licheniformis, B. mojavensis, B. toyonensis, B. anthracis, B. cereus, B. thuringiensis, and B. paranthracis were isolated and identified based on microscopic studies, biochemical tests, ribotyping, and agricultural traits. All vermi-bacteria are Gram-positive rods except Staphylococcus hominis and produce different compounds such as siderophore, indole acetic acid, catalase, oxidase, proteases, amylases, and lipases. All vermi-bacteria also act as phosphate solubilizers. Therefore, all isolated vermi-bacteria could be used as potential microbial biofertilizers to enhance crops production in Pakistan.

TP53 lacks tetramerization and N-terminal domains due to novel inactivating mutations detected in leukemia patients.

BACKGROUND: TP53 is a highly conserved tumor suppressor gene present on chromosome 17 and comprised 11 exons and 12 introns. The TP53 protein maintained the genomic integrity of the cell by regulating different pathways. The association of TP53 with leukemia and the increasing prevalence of leukemia in Pakistan instigated us to initiate the current study. MATERIALS AND METHODS: The TP53 gene of acute myeloid leukemia patients (n = 23) and normal individuals (n = 30) was amplified through polymerase chain reaction (PCR). The PCR amplified products of 3 samples 1 normal (NC-30) and 2 cancerous (LK-6 and LK-19) were subjected to deoxyribonucleic acid (DNA) sequence analysis. Bioinformatics analysis of the obtained DNA sequences helped to identify nature, type, and functional impact of mutations, if any. RESULTS: Results revealed 2 novel mutations in Case No. 1 (c. G >A10987 and c. InsA13298_13299) and Case No. 2 (c. InsC13284_13285, c. T >A13365) which generate a premature codon (ocher) at position 239 and lead to truncated TP53 protein. In Case No. 3, 16 novel mutations were identified and c. delC11093 mutation created a premature codon (opal) at 59th position. Hence, the resultant protein will lack its tetramerization and N-terminal domain required for its normal functioning. Moreover, some intronic mutations were noticed and found to have a negative impact on splicing related regulatory sequences. CONCLUSION: Results suggest the role of TP53 inactivating mutations in pathogenesis of leukemia.

Integrative analysis of multi-omics data highlighted TP53 as a potential diagnostic and prognostic biomarker of survival in breast invasive carcinoma patients.

The global incidence of breast invasive carcinoma (BRIC) has risen significantly in recent years, so it is important to identify the novel biomarkers for the early detection and treatment of BRIC. The role of the TP53 gene is well studied in the pathogenesis of BRIC but still, observations are conflicting. Therefore, this study was initiated to have a consolidated overview of TP53 contributions in the BRIC initiation and progression by analyzing its mutatome, expression variations, promoter methylation level, clinical outcome, and drug sensitivity analysis in BRIC using cBioPortal, UALCAN, KM plotter, and CCLE GDSC toolkit database. Mutatome analysis revealed that TP53 was mutated in 30 % BRIC cases and among all the noted mutations, missense and truncation mutation were noticed as the most frequent mutations and thought to be involved in the up-regulation of TP53 expression. TP53 transcription, translation, and promoter methylation levels in BRIC patients of various clinicopathological features were high relative to the normal controls. Kaplan Meier overall survival (OS) analysis revealed a good prognostic value of TP53 overexpression for the survival in BRIC patients. Moreover, TP53 overexpression was found to alter the effectiveness of various drugs used in the chemotherapy of BRIC. Collectively, our findings suggested that TP53 might be a potential diagnostic and prognostic marker for the survival in BRIC patients of various clinicopathological features.

Withdrawal Notice: TP53 Targetome: A Database of Novel Breast Cancer Biomarkers

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Overexpression of RAD50 is the Marker of Poor Prognosis and Drug Resistance in Breast Cancer Patients.

BACKGROUND: The prevalence of breast cancer is increasing at an alarming rate and thus demands exploration of the most relevant diagnostic biomarkers. RAD50 is a cancer susceptibility gene that encodes a DNA damage repairing protein. Its role in breast cancer as clinico-pathological specific biomarker has yet to be explored. OBJECTIVE: This study was aimed to investigate the RAD50 expression and its promoter's methylation level variations in breast invasive carcinoma patients having different clinico-pathological features. This study further explored the mutational spectrum of RAD50 and the correlation of its expression with the survival of patients and the effectiveness of drugs used for treatment. METHODS: Enrichment analysis of RAD50 was accomplished using the platform of GeneCards. The information regarding RAD50 expression, its promoter methylation and impact on survival of patient was retrieved from TCGA and CPTAC databases. However, the effect of RAD50 expression on tumor's response to various drugs was deduced through the analysis of CCLE and genomic of GDSC dataset. RESULTS: The promoter hyper-methylation and elevated expression of RAD50 was documented in various subgroups of breast invasive carcinoma. The subjects having low/medium expression levels were observed to survive longer than patients exhibiting high expression of RAD50 except for post-menopausal subjects. The frequency of missense mutations was higher in RAD50 than truncating mutations. Most of the drugs were found to have a positive correlation with RAD50 expression. CONCLUSION: The status of RAD50 promoter's methylation inversely correlates with the expression level of RAD50. While RAD50 is overexpressed in breast cancer patients and thus makes tumor resistant against many anti-cancer drugs.