Mechanisms of medicinal, pharmaceutical, and immunomodulatory action of probiotics bacteria and their secondary metabolites against disease management: an overview.

Probiotics or bacteriotherapy is today's hot issue for public entities (Food and Agriculture Organization, and World Health Organization) as well as health and food industries since Metchnikoff and his colleagues hypothesized the correlation between probiotic consumption and human's health. They contribute to the newest and highly efficient arena of promising biotherapeutics. These are usually attractive in biomedical applications such as gut-related diseases like irritable bowel disease, diarrhea, gastrointestinal disorders, fungal infections, various allergies, parasitic and bacterial infections, viral diseases, and intestinal inflammation, and are also worth immunomodulation. The useful impact of probiotics is not limited to gut-related diseases alone. Still, these have proven benefits in various acute and chronic infectious diseases, like cancer, human immunodeficiency virus (HIV) diseases, and high serum cholesterol. Recently, different researchers have paid special attention to investigating biomedical applications of probiotics, but consolidated data regarding bacteriotherapy with a detailed mechanistically applied approach is scarce and controversial. The present article reviews the bio-interface of probiotic strains, mainly (i) why the demand for probiotics?, (ii) the current status of probiotics, (iii) an alternative to antibiotics, (iv) the potential applications towards disease management, (v) probiotics and industrialization, and (vi) futuristic approach.

Characterization of heavy metal-associated bacteria from petroleum-contaminated soil and their resistogram and antibiogram analysis.

The aim of the current study was to screen and identify heavy metal (chromium, cadmium, and lead) associated bacteria from petroleum-contaminated soil of district Muzaffarabad, Azad Jammu and Kashmir, Pakistan to develop ecofriendly technology for contaminated soil remediation. The petroleum-contaminated soil was collected from 99 different localities of district Muzaffarabad and the detection of heavy metals via an atomic absorption spectrometer. The isolation and identification of heavy metals-associated bacteria were done via traditional and molecular methods. Resistogram and antibiogram analysis were also performed using agar well diffusion and agar disc diffusion methods. The isolated bacteria were classified into species, i.e., B. paramycoides, B. albus, B. thuringiensis, B. velezensis, B. anthracis, B. pacificus Burkholderia arboris, Burkholderia reimsis, Burkholderia aenigmatica, and Streptococcus agalactiae. All heavy metals-associated bacteria showed resistance against both high and low concentrations of chromium while sensitive towards high and low concentrations of lead in the range of 3.0 ± 0.0 mm to 13.0 ± 0.0 mm and maximum inhibition was recorded when cadmium was used. Results revealed that some bacteria showed sensitivity towards Sulphonamides, Norfloxacin, Erythromycin, and Tobramycin. It was concluded that chromium-resistant bacteria could be used as a favorable source for chromium remediation from contaminated areas and could be used as a potential microbial filter.

Green synthesised AuNps using Ajuga Bracteosa extract and AuNps-Free supernatant exhibited equivalent antibacterial and anticancerous efficacies.

The current study is designed to synthesize gold nanoparticles using Ajuga bracteosa extract, which is a highly known medicinal herb found in the northern Himalayas. The synthesized gold nanoparticles were initially characterized by UV-Vis spectrophotometer, SEM, FTIR, pXRD, and, GC-MS. Antibacterial efficacy of A. bracteosa extract, AuNps, and AuNps-free supernatant activity was checked against highly pathogenic clinical isolates of Escherichia coli, Staphylococcus aureus, and Pseudomonas aeruginosa via agar well diffusion method, assuming that supernatant might have active compounds. The Nps-free supernatant showed the maximum antibacterial activity against E. coli (20.8±0.3 mm), Staphylococcus aureus (16.5±0.5), and Pseudomonas aeruginosa (13±0.6). While green synthesized AuNps showed effective antibacterial activity (Escherichia coli (16.4±0.3mm), Staphylococcus aureus (15.05±0.5mm), and Pseudomonas aeruginosa (11.07±0.6mm)) which was high compared to A. bracteosa extract. Anticancer activity was assessed by MTT assay on U87 and HEK293 cell lines. Aj-AuNps have an antigrowth effect on both the cell lines however Aj-AuNps-free supernatant which was also evaluated along with the Aj-AuNps, showed high toxicity toward HEK293 cell line compared to U87. Further, the GC-MS analysis of supernatant showed the presence of resultant toxic compounds after the reduction of gold salt, which include Trichloromethane, Propanoic acid, 2-methyl-, methyl ester, Methyl isovalerate, Pentanoic acid, 2-hydroxy-4-methyl-, Benzene-propanoic acid, and alpha-hydroxy. Based on the observation small molecular weight ligands of Ajuga bracteosa were analyzed in-silico for their binding efficacy towards selected membrane proteins of our target pathogens. RMSD is also calculated for the best docked protein ligand pose. The results revealed that among all listed ligands, Ergosterol and Decacetylajugrin IV have high virtuous binding affinities towards the membrane proteins of targeted pathogens. The current findings revealed that the Aj-AuNps are good antibacterial as well as anticancerous agents while the Nps-free supernatant is also exceedingly effective against resistant pathogens and cancer cell lines.

Acute toxicity, anti-diabetic, and anti-cancerous potential of Trillium govanianum-conjugated silver nanoparticles in Balb/c mice.

BACKGROUND: The current study aimed to develop an economic plant-based therapeutic agent to improve the treatment strategies for diseases at the nano-scale because Cancer and Diabetes mellitus are major concerns in developing countries. Therefore, in vitro and in vivo anti-diabetic and anti-cancerous activities of Trillium govanianum conjugated silver nanoparticles were assessed. METHODS: In the current study synthesis of silver nanoparticles using Trillium govanianum and characterization were done using a scanning electron microscope, UV-visible spectrophotometer, and FTIR analysis. The in vitro and in vivo anti-diabetic and anti-cancerous potential (200 mg/kg and 400 mg/kg) were carried out. RESULTS: It was discovered that Balb/c mice did not show any major alterations during observation of acute oral toxicity when administered orally both TGaqu (1000 mg/kg) and TGAgNPs (1000 mg/kg), and results revealed that 1000 mg/kg is not lethal dose as did not find any abnormalities in epidermal and dermal layers when exposed to TGAgNPs. In vitro studies showed that TGAgNPs could not only inhibit alpha-glucosidase and protein kinases but were also potent against the brine shrimp. Though, a significant reduction in blood glucose levels and significant anti-cancerous effects was recorded when alloxan-treated and CCl4-induced mice were treated with TGAgNPs and TGaqu. CONCLUSION: Both in vivo and in vitro studies revealed that TGaqu and TGAgNPs are not toxic at 200 mg/kg, 400 mg/kg, and 1000 mg/kg doses and possess strong anti-diabetic and anti-cancerous effects due to the presence of phyto-constituents. Further, suggesting that green synthesized silver nanoparticles could be used in pharmaceutical industries to develop potent therapeutic agents.

Synthesis, Characterization, Acute Dermal Toxicity, Anti-inflammatory, and Wound Healing Potential of Biogenic Silver Nanoparticles in Balb C Mice.

AIM: The current study aimed to develop an economic plant-based therapeutic agent to improve the treatment strategies for diseases at the nano-scale. METHOD: In the current research, silver nanoparticles were synthesized using Trillium govanianum aqueous extract. Characterizations were done using UV-Visible spectrophotometer, X-ray diffraction, scanning electron microscopy, and Fourier transform infrared spectroscopy. In vivo biological activities such as acute dermal toxicity, wound healing, and anti-inflammatory were done on Balb C mice. Absorbance at 295 nm corresponds to the out-of-plane quadrupole Plasmon-resonance while at 350 nm corresponds to in-plane dipole resonance. SEM images showed the morphology of TGAgNPs is not exactly spherical while XRD analysis shows that highly crystalline TGAgNPs with an average size of 27.94 nm. The FTIR spectrum represents sharp peaks of aldehyde, amide I, aromatic rings, and polysaccharides. The microscopic assessment did not find any epidermal and dermal layer abnormalities in Blab C mice when exposed to TGAgNPs during acute dermal toxicity. RESULT & DISCUSSION: Results revealed that 1000 mg/kg is not a lethal dose. In the wound healing activity, no mortality and no abnormal signs were observed when petroleum jelly, Nitrofuranose, TGaqu, and TGAgNPs-based ointments were applied. Enhanced epithelization was recorded in TGaqu and TGAgNPs treated mice (p≤0.001). The wound contraction percentage was higher in nitrofuranose-treated mice (74%) followed by TGAgNPs (71%), and TGaqu (69%) compared to vehicle-treated and open-wounded mice. The paw edema model proved the potential use of TGAgNPs and TGaqu as anti-inflammatory agents. CONCLUSION: Hence, the results proved that both TGaqu and TGAgNPs are not toxic and possessed strong anti-inflammatory and wound-healing effects due to the presence of phytochemical constituents and could be used in various drug production as a therapeutic agent.

New Earthworm Record from Division Muzaffarabad, Azad Kashmir, Pakistan Supported by Molecular Markers.

Earthworm diversity and ecology in Pakistan is poorly known, especially in the region of Azad Jammu & Kashmir. An earthworm community survey assisted by genetic barcoding detected an unidentified species which could constitute a new record for Pakistan. Morphological study revealed its identity as Perelia kaznakovi. Additionally, Bayesian phylogenetic inference based on five mitochondrial and nuclear molecular markers was performed. Results provided a phylogenetic placement of the genus Perelia within Lumbricidae for the first time, indicating a close relationship with Eophila. This approach should be implemented to Perelia arnoldiana and further representatives of the genus in order to understand their biogeography, diversity and evolutionary history.

Cow dung putrefaction via vermicomposting using Eisenia fetida and its influence on seed sprouting and vegetative growth of Viola wittrockiana (pansy).

The current research was conducted at Vermi-tech Unit, Muzaffarabad in 2018 to evaluate the efficacy of cow dung and vermicompost on seed sprouting, seedlings, and vegetative developmental parameters of Viola x wittrokiana (pansy). In the current study, vermicompost was produced using Eisenia fetida. Physicochemical parameters of vermicompost and organic manure were recorded before each experimentation. The potting experiment was designed and comprised of eight germination mediums containing different combinations of soil, sand, cow dung, and various concentrations of vermicompost such as 10% VC, 15% VC, 20% VC, 25% VC, 30% VC, and 35% VC. Seed sprouting and seedling developmental parameters were observed for 28 days while vegetative plant growth parameters were recorded after 10 weeks of transplantation in various vermicompost amended germination media. Pre and post-physicochemical analysis of germination media were also recorded to check their quality and permanency. The current findings showed that 30% VC germination media was an effective dose for early seed germination initiation and all seed germination parameters. However, the significant vegetative plant growth and flowering parameters of pansy occurred at 35% VC. Findings revealed that vermicompost not only enhanced the seed germination and growth of pansy but also improved soil health. These results indicate that vermicompost can be exploited as a potent bio-fertilizer for ornamental plant production.

Bioremediation of heavy metals polluted environment and decolourization of black liquor using microbial biofilms.

BACKGROUND: With increased urbanization and industrialization, modern life has led to an anthropogenic impact on the biosphere. Heavy metals pollution and pollutants from black liquor (BL) have caused severe effects on environment and living organisms. Bacterial biofilm has potential to remediate heavy metals and remove BL from the environment. Hence, this study was planned to investigate the potential of microbial biofilms for the bioremediation of heavy metals and BL polluted environments. METHODS AND RESULTS: Eleven biofilm forming bacterial strains (SB1, SB2, SC1, AF1, 5A, BC-1, BC-2, BC-3, BC-4, BC-5 and BC-6) were isolated and identified upto species level via 16S rRNA gene sequencing. Biofilm strains belonging to Bacillus and Lysinibacillus sphaericus were used to remediate heavy metals (Pb, Ni, Mn, Zn, Cu, and Co). Atomic absorption spectroscopy showed significantly high (P ≤ 0.05) bioremediation potential by L. sphaericus biofilm (1462.0 ± 0.67 µgmL-1) against zinc (Zn). Similarly, Pseudomonas putida biofilm significantly (P ≤ 0.05) decolourized (65.1%) BL. Fourier transform infrared (FTIR) analysis of treated heavy metals showed the shifting of major peaks (1637 & 1629-1647, 1633 & 1635-1643, and 1638-1633 cm-1) corresponding to specific amide groups due to C = O stretching. CONCLUSION: The study suggested that biofilm of the microbial flora from tanneries and pulp paper effluents possesses a strong potential for heavy metals bioremediation and BL decolourization. To our knowledge, this is the first report showing promising biofilm remediation potential of bacterial flora of tanneries and pulp-paper effluent from Kasur and Sheikhupura, Punjab, Pakistan, against heavy metals and BL.

Interactions of Chitosan-coated Green Synthesized Silver Nanoparticles using Mentha spicata and Standard Antibiotics against Bacterial Pathogens.

BACKGROUND: Infectious diseases are caused by various multidrug-resistant pathogenic bacteria and in recent scenarios, nanoparticles have been used as innovative antimicrobial agents. AIMS: This current research aimed to evaluate the bactericidal effect of chitosan-coated green synthesized silver nanoparticles using aqueous extract of Mentha spicata (MSaqu) against bacterial pathogens, i.e., Pseudomonas aeruginosa, Escherichia coli, Klebsiella pneumoniae, Pseudomonas aeruginosa, Serratia marcescens, Staphylococcus aureus, and Streptococcus pyogenes. METHODS: Synthesis and characterization of silver nanoparticles (MSAgNPs) were carried out via atomic absorption spectrometer and Fourier-transform infrared spectroscopy. Agar well and agar disc diffusion methods were used to assess the antibacterial and synergistic effect of chitosanmediated biogenic silver nanoparticles and standard antibiotics. Three types of interactions, i.e., antagonistic (↓), synergistic (↑), and additive (¥) were observed. RESULTS: Synergistic effect was recorded against Pseudomonas aeruginosa (8.5±0.25 mm↑), Serratia marcescens (19.0±1.0 mm↑), and Klebsiela pneumonia (8.5±0.25 mm↑), an additive effect was exhibited by Escherichia coli (9.0±0.0 mm¥), Streptococcus pyogenes (10.0±0.0 mm¥), and Staphylococcus aureus (7.5±0.25 mm↓) and they showed antagonistic effects when chitosan-coated silver nanoparticles (CLMSAgNPs) were applied compared to chitosan, MSaqu, and MSAgNPs. Interesting antibacterial results were recorded when chitosan-coated Mentha spicata extract and silver nanoparticles were applied along with antibiotics. The synergistic effects of chitosan-coated silver nanoparticles (CLMSAgNPs) + K were recorded against E. coli (14.5±0.25 mm). The synergistic effects of chitosan-coated silver nanoparticles (CLMSAgNPs) + AML were recorded against E. coli (5.5±0.0 mm), S. pyogenes (10.0±0.0 mm), K. pneumonia (5.5±0.0 mm), and S. aureus (4.0±0.0 mm). The synergistic effects of chitosan-coated silver nanoparticles (CLMSAgNPs) + NOR were recorded against E. coli (16.0±0.0 mm), P. aeruginosa (19.0±0.0 mm), S. marcescens (19.5±0.25 mm), S. pyogenes (11.5.0±0.25 mm), K. pneumonia (23.0±0.0 mm), and S. aureus (8.5±0.25 mm). CONCLUSION: Current findings concluded that chitosan-coated biogenic silver nanoparticles have potential bactericidal effects against infectious pathogens and could be used as forthcoming antibacterial agents.

Efficacy of cow and buffalo dung on vermiremediation and phytoremediation of heavy metals via Fourier-transform infrared spectroscopy and comet assay.

Heavy metal contamination raised significant concerns throughout the world. The current research aimed to evaluate the impact of organic manure (cow dung and buffalo dung) on vermiremediation and phytoremediation and to remediate heavy metals, i.e., cadmium, lead, and chromium, from artificial contaminated soil via both remediation techniques. The impact of livestock manure was evaluated for the first time which could be effective in in situ as well as ex situ studies. Eisenia fetida, Pheretima lignicola, and Spinacia oleracea were used for the remediation process. Results revealed that E. fetida tolerated lead at 280 mg, cadmium at 150 mg, and chromium at 860 mg compared to P. lignicola. The growth and reproduction of E. fetida were efficient in the cow dung manure compared to buffalo dung. Similarly, seed germination and growth of Spinacia oleracea were better in cow dung media compared to buffalo dung. Bioaccumulation factor showed that E. fetida showed a higher accumulation of heavy metals in their tissues when vermi + phytoremediation was jointly applied (9.50 mg/l of Pb, 24.166 of Cd, and 6.695 of Cr). Fourier-transform infrared spectroscopy indicated that heavy metals had no drastic effects on E. fetida and S. oleracea. Similarly, comet assay revealed that heavy metals had no genotoxic effect on the E. fetida and S. oleracea. It was concluded that both E. fetida and S. oleracea are appropriate for heavy metals remediation in cow dung manure.

COYOTE: Sequence-derived structural descriptors-based computational identification of glycoproteins.

Glycoproteins play an important and ubiquitous role in many biological processes such as protein folding, cell-to-cell signaling, invading microorganism infection, tumor metastasis, and leukocyte trafficking. The key mechanism of glycoproteins must be revealed to model and refine glycosylated protein recognition, which will eventually assist in the design and discovery of carbohydrate-derived therapeutics. Experimental procedures involving wet-lab experiments to reveal glycoproteins are very time-consuming, laborious, and highly costly. However, costly and tedious experimental procedures can be assisted by ranking the most probable glycoproteins through computational methods with improved accuracy. In this study, we have proposed a novel machine learning-based predictive model for glycoproteins identification. Our proposed model is based on sequence-derived structural descriptors (SDSD) that fill the gap of unavailability of protein 3D structures and lack of accuracy in sequence information alone. Through a series of simulation studies, we have shown that our proposed model gives state-of-the-art generalization performance verified through various machine learning-centric and biologically relevant techniques and metrics. Through data mining in this study, we have also identified the role of descriptors in determining glycoproteins. Python-based standalone code together with a webserver implementation of our proposed model (COYOTE: identifiCation Of glYcoprOteins Through sEquences) is available at the URL: https://sites.google.com/view/wajidarshad/software.

Molecular Characterization and Phylogenetic Analysis of Earthworm Species Collected from Different Soil Habitats of Poonch Division Azad Jammu and Kashmir Pakistan.

This study aims to analyze molecular characterization and phylogenetic analysis of earthworm species collected from different soil habitats of Poonch division Azad Kashmir Pakistan by using CO1 gene partial sequencing methodology. Samples gathered randomly from 18 study sites (127 localities) by digging and hand sorting methods were preserved in pure ethanol at -20°C. The modified CTAB (Cetyltrimethyl ammonium bromide) method extracted high quality DNA from region of representative earthworm's caudal region. This extracted DNA was used to amplify the 700 bp partial region of the cytochrome oxidase I (COI) gene with LCO1490 and HCO2198 universal primers. All of the obtained amplified gene sequences were aligned, edited and analyzed using MEGA X software to characterize different species of earthworms. Thirty-eight (38) Barcoding sequences belonging to 11 different strains of earthworms were successfully generated. Their phylogenetic analysis revealed that 7 Barcoding sequences gave maximum similarity with the available online database, while the rest of the 4 sequences gave lower similarity than the maximum threshold level. The collected DNA barcode sequences were also clustered together by the maximum likelihood method and the resultant phylogenetic tree revealed they belong to different family lineages. Moreover the identified earthworm species have a close evolutionary link with the earthworm fauna of south and central Asia instead of Europe, which might be due to similar climate of both regions.

Screening of Antibacterial Efficacy of Chitosan Encapsulated Probiotics (Lactococcus lactis and Lactobacillus curvattus) against Clinical Bacterial Pathogens.

Probiotics frontier in depressing the clinical bacterial pathogens to avoid multidrug resistance phenomenon. The present study aimed to determine the antibacterial efficiency of chitosan encapsulated probiotics isolated from buffalo milk samples against clinical bacterial pathogens. The Agar well method was used for antibacterial activity. Lactococcus lactis (A) and Lactobacillus curvattus (B) were isolated from fresh buffalo milk samples, identified via culturing media, Gram's staining, biochemical tests, and antibiogram analysis. Encapsulation of probiotics was carried out using chitosan and was characterized via a scanning electron microscope. Antibiogram analysis elicit that L. lactis culture (A1) was highly sensitive to chloramphenicol (17.66±0.47 mm), tobramycin (15.33±0.47 mm), and ciprofloxacin (12.33±0.47 mm) and resistant against tetracycline, Penicillin G, Erythromycin, Amoxycillin, Ceftriaxone, Cephalothin, and Cephradine, while L. curvattus culture (B1) was affected by Ceftriaxone (18.67±0.47 mm), Amoxycillin (14.33±0.94 mm), Cephalothin (13.67±0.47 mm), Erythromycin (13.33±0.47 mm), Penicillin G (12.67±0.47 mm), Cephradine (10.33±0.47 mm), and Chloramphenicol (9.67±0.47 mm) and resistant against tetracycline, Tobramycin, and Ciprofloxacin. Antibacterial efficacy of non-encapsulated probiotic cultures was significant and maximum inhibition of bacterial were recorded compared to their cellular components. SEM of encapsulated probiotics revealed that they were successfully covered with a chitosan protective layer and could be effective as bio-preservatives due to being slowly released at the target site. The current study concluded that L. lactis, L. curvattus, and their cellular components have a significant bactericidal effect against infectious pathogens and could be used as a potential therapeutic drug against infectious diseases.

In vitro Antimicrobial Activity of Biogenically Synthesized Nickel and Zinc Nanoparticles against Selected Pathogenic Bacterial Strains.

Bacterial resistance to already present antibiotics demands for new approaches in field of medicine. Scientists prefer nanoparticles (NPs) due to their promising potential in many applications. Two bacterial strains, Escherichia coli and Bacillus subtilis were used for biogenic synthesis of NPs. Characterization of prepared NPs was accomplished using UV-vis spectroscopy and fourier transform infrared spectroscopy (FTIR). The prepared NPs were confirmed by the color change from pale yellow to having white deposition for Zn NPs while from dark green to light green for Ni NPs. UV-vis spectroscopy of E. coli and B. subtilis based ZnNPs showed highest peak at 354nm and 362nm, respectively. Likewise, E. coli and B. subtilis NiNPs showed peaks at 246 nm and 238 nm, respectively. Antibacterial activity of B. subtilis based ZnNPs showed significant (p ≤ 0.05) zone of inhibition (ZOI; 27.3±0.6) against B. subtilis and 26.66±0.67 against E. coli at 100 mg/mL. Antibacterial activity of E. coli based ZnNPs showed 8.3±0.3 ZOI against B. subtilis and 6.6±0.3 ZOI against E. coli while NiNPs showed (25.0±0.0 mm) (ZOI) against B. subtilis and (25.0 ± 0.3 mm) against E. coli. Minimum inhibitory concentration (MIC) of E. coli ZnNPs showed values of 6.7±0.3 µg/mL for E. coli and 4.7±0.3 µg/mL for B. subtilis. MIC of B. subtilis ZnNPs showed 5.3±0.3 µg/mL for E. coli and 6.6±0.3 µg/mL for B. subtilis while NiNPs showed 33.0±1.0 µg/mL against E. coli and 24.0±1.0 µg/mL against B. subtilis as effective inhibitory concentrations. Minimum bactericidal concentration (MBC) of E. coli ZnNPs showed 7.3±0.3 µg/mL for E. coli and 8.3±0.3 µg/mL for B. subtilis. MBC of B. subtilis ZnNPs showed 7.6±0.3 µg/mL for E. coli and 8.6±0.3 µg/mL for B. subtilis while NiNPs showed 45.7±1.3 µg/mL against E. coli and 33.0±1.0 µg/mL against B. subtilis as effective inhibitory concentrations. It was concluded from the current study that biogenically synthesized ZnNPs and NiNPs are effective as promising antibacterial agents and have potential applications in biomedical fields.

Glycol-Chitosan-Based Technetium-99m-Loaded Multifunctional Nanomicelles: Synthesis, Evaluation, and In Vivo Biodistribution.

We hereby propose the use of stable, biocompatible, and uniformly sized polymeric micelles as high-radiotracer-payload carriers at region-of-interest with negligible background activity due to no or low offsite radiolysis. We modified glycol chitosan (GC) polymer with varying levels of palmitoylation (P) and quaternization (Q). Quaternary ammonium palmitoyl glycol chitosan (GCPQ) with a Q:P ratio of 9:35 (Q9P35GC) offers >99% biocompatibility at 10 mg mL−1. Q9P35GC micelles exhibit >99% 99mTechnetium (99mTc) radiolabeling via the stannous chloride reduction method without heat. The 99mTc-Q9P35GC micelles (65 ± 3 nm) exhibit >98% 6 h serum stability at 37 °C and 7 day of radiochemical stability at 25 °C. HepG2 cells show a higher uptake of FITC-Q9P35GC than Q13P15GC and Q20P15GC. The in vivo 24 h organ cumulated activity (MBq h) order follows: liver (234.4) > kidneys (60.95) > GIT (0.73) > spleen (88.84). The liver to organ ratio remains higher than 2.4, rendering a better contrast in the liver. The radiotracer uptake decreases significantly in fibrotic vs. normal liver, whereas a blocking study with excess Q9P35GC significantly decreases the radiotracer uptake in a healthy vs. fibrotic liver. FITC-Q9P35GC shows in vivo hepato-specific uptake. Radiotracer liver uptake profile follows reversible binding kinetics with data fitting to two-tissue compartmental (2T), and graphical Ichise multilinear analysis (MA2) with lower AIC and higher R2 values, respectively. The study concludes that 99mTc-Q9P35GC can be a robust radiotracer for noninvasive hepatocyte function assessment and diagnosis of liver fibrosis. Furthermore, its multifunctional properties enable it to be a promising platform for nanotheranostic applications.

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.

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.

A Comparative Analysis of Antimicrobial, Antibiofilm and Antioxidant Activity of Silver Nanoparticles Synthesized from Erythrina Suberosa Roxb. and Ceiba Pentandra.

Biofilm forming bacteria can cause serious health problems that are difficult to combat. Silver nanoparticles (Ag-NPs) synthesized from plant extracts have potential to fight and eradicate biofilmforming bacteria. In the present research, AgNPs were synthesized using leaf and bark extract of Erythrina suberosa Roxb. and Ceiba pentandra L. and their antibiofilm, antioxidant and antibacterial activity was checked. Phytochemical analysis of the plant extracts showed important bioactive compounds such as tannins, saponins, steroids, phenolics, alkaloids, flavonoids and glycosides. The AgNPs were synthesized and confirmed by visual color observation and UV-Vis spectrophotometer. Visual color observation showed that the color of the leaf and bark extracts of E. suberosa and C. pentandra turned into brown. UV-Vis spectra analysis showed absorbance peak range between 430-450 nm. The antioxidant activity of the AgNPs was determined by FRAC (Ferrous reducing antioxidant capacity) assay. Synthesized AgNPs from all sources showed significant antioxidant activity. However, antioxidant activity of E. suberosa AgNPs was significant compared to other sources. Antibacterial activity and biofilm forming assay was analyzed against Escherichia coli, Pseudomonas aeruginosa and Staphylococcus aureus. The synthesized AgNPs silver nanoparticles showed significant (p ≤ 0.05) antibacterial activity against all the bacteria. The maximum zone of inhibition was found in case of E. suberosa AgNPs bark extract against P. aeruginosa was 20±1.154 mm. The results of biofilm forming assay showed that the AgNPs from all sources significantly (p ≤ 0.05) inhibited the activity of biofilms by all the tested bacteria. From results, it can be concluded that AgNPs synthesized from both plants can be used in developing antimicrobial compounds.

SIP: A computational prediction of S-Adenosyl methionine (SAM) interacting proteins and their interaction sites through primary structures.

S-Adenosyl methionine (SAM), a universal methyl group donor, plays a vital role in biosynthesis and acts as an inhibitor to many enzymes. Due to protein interaction-dependent biological role, SAM has become a favorite target in various therapeutical and clinical studies such as treating cancer, Alzheimer's, epilepsy, and neurological disorders. Therefore, the identification of the SAM interacting proteins and their interaction sites is a biologically significant problem. However, wet-lab techniques, though accurate, to identify SAM interactions and interaction sites are tedious and costly. Therefore, efficient and accurate computational methods for this purpose are vital to the design and assist such wet-lab experiments. In this study, we present machine learning-based models to predict SAM interacting proteins and their interaction sites by using only primary structures of proteins. Here we modeled SAM interaction prediction through whole protein sequence features along with different classifiers. Whereas, we modeled SAM interaction site prediction through overlapping sequence windows and ranking with multiple instance learning that allows handling imprecisely annotated SAM interaction sites. Through a series of simulation studies along with biological significant evaluation, we showed that our proposed models give a state-of-the-art performance for both SAM interaction and interaction site prediction. Through data mining in this study, we have also identified various characteristics of amino acid sub-sequences and their relative position to effectively locate interaction sites in a SAM interacting protein. Python code for training and evaluating our proposed models together with a webserver implementation as SIP (Sam Interaction Predictor) is available at the URL: https://sites.google.com/view/wajidarshad/software.

Biogenic Synthesis, Characterization and Antibacterial Properties of Silver Nanoparticles against Human Pathogens.

Biogenic synthesis of silver nanoparticles (AgNPs) is more eco-friendly and cost-effective approach as compared to the conventional chemical synthesis. Biologically synthesized AgNPs have been proved as therapeutically effective and valuable compounds. In this study, the four bacterial strains Escherichia coli (MT448673), Pseudomonas aeruginosa (MN900691), Bacillus subtilis (MN900684) and Bacillus licheniformis (MN900686) were used for the biogenic synthesis of AgNPs. Agar well diffusion assay revealed to determine the antibacterial activity of all biogenically synthesized AGNPs showed that P. aeruginosa AgNPs possessed significantly high (p < 0.05) antibacterial potential against all tested isolates. The one-way ANOVA test showed that that P. aeruginosa AgNPs showed significantly (p < 0.05) larger zones of inhibition (ZOI: 19 to 22 mm) compared to the positive control (rifampicin: 50 µg/mL) while no ZOI was observed against negative control (Dimethyl sulfoxide: DMSO). Minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) concentration against four test strains also showed that among all biogenically synthesized NPs, P. aeruginosa AgNPs showed effective MIC (3.3-3.6 µg/mL) and MBC (4.3-4.6 µg/mL). Hence, P. aeruginosa AGNPs were characterized using visual UV vis-spectroscopy, X-ray diffractometer (XRD), fourier transform infrared (FTIR) and scanning electron microscopy (SEM). The formation of peak around 430 nm indicated the formation of AgNPs while the FTIR confirmed the involvement of biological molecules in the formation of nanoparticles (NPs). SEM revealed that the NPs were of approximately 40 nm. Overall, this study suggested that the biogenically synthesized nanoparticles could be utilized as effective antimicrobial agents for effective disease control.