Anti-Arthritic and Immunomodulatory Potential of Methanolic, n-Hexane, and Ethyl Acetate Fractions of Bark of Acacia modesta on Complete Freund's Adjuvant-Induced Arthritis in Rats.

Rheumatoid arthritis is an autoimmune disorder and topic of interest for researchers due to its increasing frequency and limited treatment. Acacia modesta Wall is known to treat rheumatic disorders in the traditional system of medicinal plants. Traditional medicines are still required for the treatment of this disease due to the large number of side-effects caused by commercial medicines. In the current study, the antiarthritic potential of methanolic extract (AM-metha), n-hexane (AM-hexa) fraction, and ethyl acetate (AM-etha) fraction of the bark of A. modesta against a complete Freund's adjuvant rat model was evaluated. Evaluation using a digital plethysmometer, macroscopic evaluation, and histopathological evaluation were conducted to determine the paw volume and arthritic scoring. ELISA was performed to assess the PGE2 levels. RT-PCR was used to evaluate the expression levels of MMP2, MMP3, MMP9, NF-κB, IL6, IL1ß, TNFα, and VEGF. Biochemical and hematological analyses were also conducted. GC/MS was also carried out to analyze the presence of medicinal compounds. The data revealed a marked reduction in the paw volume, arthritic scoring, and histopathological parameters, indicating the anti-arthritic potential of the plant. Treatment with plant extracts and fractions markedly down-regulated MMP2, MMP3, MMP9, NF-κB, IL6, IL1ß, TNFα, and VEGF levels. Similarly, PGE2 levels were also found to be ameliorated in the treatment groups, indicating the immunomodulatory property of plant bark. Plant treatment nearly normalized hematological parameters such as counts of WBCs, RBCs, and platelets, along with Hb content, thereby validating the anti-arthritic activity. GC/MS analysis disclosed the presence of strong anti-inflammatory compounds such as lupeol, oleic acid, and squalene. The study showed that A. modesta possesses anti-arthritic and immunomodulatory potential linked to significant down-regulation of pro-inflammatory and inflammatory biomarkers.

Maximizing the extraction yield of plant gum exudate using response surface methodology and artificial neural networking and pharmacological characterization.

Prunus armeniaca gum is used as food additive and ethno medicinal purpose. Two empirical models response surface methodology and artificial neural network were used to search for optimized extraction parameters for gum extraction. A four-factor design was implemented for optimization of extraction process for maximum yield which was obtained under the optimized extraction parameter (temperature, pH, extraction time, and gum/water ratio). Micro and macro-elemental composition of gum was determined by using laser induced breakdown spectroscopy. Gum was evaluated for toxicological effect and pharmacological properties. The maximum predicted yield obtained by response surface methodology and artificial neural network was 30.44 and 30.70% which was very close to maximum experimental yield 30.23%. Laser induced breakdown spectroscopic spectra confirmed the presence Calcium, Potassium, Magnesium, Sodium, Lithium, Carbon, Hydrogen, Nitrogen and Oxygen. Acute oral toxicity study showed that gum is non-toxic up to 2000 mg/Kg body weight in rabbits, accompanied by high cytotoxic effects of gum against HepG2 and MCF-7cells by MTT assay. Overall, Aqueous solution of gum showed various pharmacological activities with significant value of antioxidant, antibacterial, anti-nociceptive, anti-cancer, anti-inflammatory and thrombolytic activities. Thus, optimization of parameters using mathematical models cans offer better prediction and estimations with enhanced pharmacological properties of extracted components.

Green Synthesis: The Antibacterial and Photocatalytic Potential of Silver Nanoparticles Using Extract of Teucrium stocksianum.

Green synthesis is one of the promising pathways for biologically active nanoscale materials. Herein, an eco-friendly synthesis of silver nanoparticles (SNPs) was carried out using an extract of Teucrium stocksianum. The biological reduction and size of NPS were optimized by controlling the physicochemical parameters such as concentration, temperature, and pH. A comparison of fresh and air-dried plant extracts was also undertaken to establish a reproducible methodology. The biosynthesized SNPs were characterized by UV-Vis spectroscopy, FT-IR, SEM, DLS, and XRD analyses. The prepared SNPs exhibited significant biological potential against multi-drug-resistant pathogenic strains. The results revealed that the biosynthesized SNPs exhibit high antimicrobial activity at low concentrations compared to the parent plant extract. Minimum inhibition concentration (MIC) values were found between 5.3 µg/mL to 9.7 µg/mL for the biosynthesized SNPs, whereas the aqueous extract of the plant showed many high values of MIC, i.e., between 69 and 98 µg/ML. Furthermore, the synthesized SNPs were found efficient in the photolytic degradation of methylene blue under sunlight.

A green and environmental sustainable approach to synthesis the Mn oxide nanomaterial from Punica granatum leaf extracts and its in vitro biological applications.

Pathogenic fungal infections in fruit cause economic losses and have deleterious effects on human health globally. Despite the low pH and high water contents of vegetables and fresh, ripened fruits, they are prone to fungal and bacterial diseases. The ever-increasing resistance of phytopathogens toward pesticides, fungicides and bactericides has resulted in substantial threats to plant growth and production in recent years. However, plant-mediated nanoparticles are useful tools for combating parasitic fungi and bacteria. Herein, we synthesized biogenic manganese oxide nanoparticles (MnONPs) from an extract of Punica granatum (P. granatum), and these nanoparticles showed significant antifungal and antibacterial activities. The production of MnONPs from plant extracts was confirmed by infrared spectroscopy (FTIR), X-ray diffraction (XRD) and UV visible spectroscopy (UV). The surface morphology and shape of the nanoparticles were characterized by scanning electron microscopy (SEM) and transmission electron microscopy (TEM). Using a detached fruit method, the MnONPs were shown to exhibit significant antimicrobial activities against two bacterial strains, E. coli and S. aureus, and against the fungal species P. digitatum. The results revealed that the MnONPs had a minimum antimicrobial activity at 25 µg/mL and a maximum antimicrobial activity at 100 µg/mL against bacterial strains in lemon (citrus). Furthermore, the MnONPs exhibited significant ROS scavenging activity. Finally, inconclusive results from the green-synthesized MnONPs magnified their significant synergetic effects on the shelf life of tomatoes (Lycopercicum esculantum) and indicated that they could be used to counteract the phytopathological effects of postharvest fungal diseases in fruits and vegetables. Overall, this method of MnONPs synthesis is inexpensive, rapid and ecofriendly. MnONPs can be used as potential antimicrobial agents against different microbial species.

Evaluation of di-rhamnolipid biosurfactants production by a novel Pseudomonas sp. S1WB: Optimization, characterization and effect on petroleum-hydrocarbon degradation.

Rhamnolipid biosurfactants are multifunctional compounds that can play an indispensable role in biotechnological, biomedical, and environmental bioremediation-related fields, and have attracted significant attention in recent years. Herein, a novel strain Pseudomonas sp. S1WB was isolated from an oil-contaminated water sample. The biosurfactants produced by this strain have capabilities to reduce surface tension (SFT) at 32.75 ± 1.63 mN/m and emulsified 50.2 ± 1.13 % in liquid media containing 1 % used engine oil (UEO) as the sole carbon source. However, the lowest SFT reduction (28.25 ± 0.21), highest emulsification index (60.15 ± 0.07), and the maximum yields (900 mg/L) were achieved under optimized conditions; where, the glucose/urea and glycerol/urea combinations were found efficient carbon and nitrogen substrates for improved biosurfactants production. Biosurfactants product was characterized using ultra-high performance liquid chromatography-mass spectrometry (UHPLC- MS) and detected various di- rhamnolipids congeners. In addition, the di-rhamnolipids produced by S1WB strain was found highly stable in terms of surface activity and EI indices at different environmental factors i.e. temperature, pH and various NaCl concentrations, where, emulsifying property was found high stable till 30 days of incubation. Moreover, the stain was capable to degrade hydrocarbon at 42.2 ± 0.04 %, and the Gas chromatography- mass spectrometry (GC-MS) profile showed the majority of peak intensities of hydrocarbons have been completely degraded compared to control.

Controllable synthesis and stabilization of Tamarix aphylla-mediated copper oxide nanoparticles for the management of Fusarium wilt on musk melon.

Excessive use of pesticides and mineral fertilizers poses a serious threat to ecoenvironment sustainability and human health. Nano pesticides or Nano fungicides have attained great attention in the field of agriculture due to their unique characteristics, by improving crop growth with enhancing pathogenesis-related defense system. However, there is a need to develop a sustainable mechanism for the synthesis of fungicides which replace the chemical pesticides to avoid their hazardous impact. Here in, Tamarix aphylla mediated CuO-Nanoparticles (NPs) were synthesized, characterized and their activity was evaluated under in-vitro and in-vivo conditions. The structural and elemental analysis of NPs were carried out by using X-ray powder diffraction (XRD), Fourier transformed infrared spectroscopy (FTIR), UV-visible spectrophotometer, Scanning electron microscope (SEM) and Transmission electron microscope (TEM). In the greenhouse, at an optimum concentration of 50 mg/L reduced disease severity very effectively and enhanced plant growth. Application of NPs also assisted in the induction of systemic response of defense-related genes in melon. Under In vitro condition at 100 mg/L significantly reduced mycelial growth (84.5%) by directly acting on the pathogenic cell wall. Our work confirmed that dosedependent concentration of T. aphylla extract based biological CuO-NPs enhance plant growth and help to effectively resist against F. oxysporum infection. Supplementary Information: The online version contains supplementary material available at 10.1007/s13205-022-03189-0.

Effect of natural microbiome and culturable biosurfactants-producing bacterial consortia of freshwater lake on petroleum-hydrocarbon degradation.

Freshwater lake ecosystem is a reservior of valuable microbial diversity. It needs to be explored for addressing key environmental issues like petroleum-hydrocarbon contamination. In this work, the microbial communities (pre and post enriched with petroleum-hydrocarbons) from different layers of freshwater lake, i.e. surface water, sediments and deepwater, were explored through metagenomic and culture-dependent approaches. A total of 41 bacterial phyla were retrieved from pre-enriched samples, which were significantly reduced in enriched samples where Proteobacteria were dominant (87% to 100%) followed by Bacteroidetes (7.37%) and Verrucomicrobia (3.06%). The most dominant hydrocarbon-degrading genera were extensively verified as Pseudomonas (48.65%), Acinetobacter (45.38%), Stenotrophomonas (3.16%) and Brevundimonas (2.07%) in surface water (S1WCC); Acinetobacter (62.46%), Aeromonas (10.7%), Sphingobacterium (5.20%) and Pseudomonas (4.23%) in sediment (S2MCC); and Acinetobacter (46.57%), Pseudomonas (13.10%), Comamonas (12.93%), Flavobacterium (12.18%) and Enterobacter (9.62%) in deep water (S4WCC). Additionally, the maximum biodegradation of petroleum-hydrocarbons (i.e. used engine oil or UEO) was achieved by microbiome of S2MCC (67.60 ± 0.08%) followed by S4WCC (59.70 ± 0.12%), whereas only 36.80 ± 0.10% degradation was achieved by S1WCC microbiome. On the other hand, UEO degradation by cultivable biosurfactant-producing single cultures such as Pseudomonas sp. S2WE, Pseudomonas sp. S2WG, Pseudomonas sp. S2MS, Ochrobactrum sp. S1MM and Bacillus nealsonii S2MT showed 31.10 ± 0.08% to 40.50 ± 0.11% biodegradation. Comparatively, the biodegradation efficiency was found higher (i.e. 42.20 ± 0.12% to 56.10 ± 0.12%) in each consortia comprising of two, three, four, and five bacterial cultures. Conclusively, the isolated culturable biosurfactants-producing bacterial consortium of freshwater lake demonstrated >80% contribution in the total petroleum-hydrocarbons degradation by the natural microbiome of the ecosystem.

Enhanced Oil Recovery by Potential Biosurfactant-Producing Halo-thermotolerant Bacteria Using Soil Washing and Sand-Packed Glass Column Techniques.

Biosurfactants offer numerous advantages over the chemical surfactants, especially in energy and environment-related applications. Microbial enhanced oil recovery (MEOR) is a technique to recover oil from reservoirs by using microbes and their metabolites. In present study, total sixteen morphologically distinct bacterial strains isolated from different salty areas of the district Khairpur Mir's, Pakistan, were investigated for their MEOR potential. Screening assays for thermotolerance and halotolerance declared 7 out of 16 (43.75%) bacterial isolates as thermotolerant (capable of growing in the temperature range 60-70 °C) and halotolerant (tolerating NaCl concentrations up to 17%, w/v). Moreover, five of them were screened as biosurfactant producers. Among, the lowest surface tension reduction was achieved with biosurfactants produced by the strains KJ2MO (27.8 mN/m) and KJ2SK (29.3 mN/m). The biosurfactant activity was found stable at temperature (100-121 °C, 1 h) and pH (4-10). Moreover, maximum oil recovery was obtained with biosurfactant of bacterial strain KJ2MO (54.7%, 51.25%) followed by KJ2SK (44.7%, 40.5%), KJ1WB (37%, 35.5%) and KJ2MD (37.8%, 31.9%) by using either techniques, i.e., soil washing and sand-packed column, respectively. Moreover, the potent species were identified as Pseudomonas pseudoalcaligenes KJ1WB, Bacillus aerius KJ2MD, Bacillus licheniformis KJ2SK, and Bacillus subtilis KJ2MO using 16S rRNA ribo-typing. The investigated species were found to be promising biosurfactants producers having potential for enhanced oil recovery and could be used in other environmental applications like bioremediation.

Determination of antimicrobial and antioxidant potential of agro-waste peels.

Natural products from plants are pulling in more interest in exploration due to their therapeutic properties like, mainly because of the drug-resistance in microbes against synthetic drugs. Thus, the present study was designed to assess the antimicrobial and antioxidant actions of some Agro-waste (garlic, ginger, onion, potato) peel extracts. Ginger peel extracts exhibited the highest FRAP (0.273±0.044 mm Fe2+ Eq/g dry weight) while garlic peel extracts exhibited the highest TAC (0.47±0.0452 mm AAE/g dry weight) values among all selections. The antimicrobial activity of the peels was evaluated against different pathogenic bacterial and fungal strains i.e.; Escherichia coli, Bacillus megaterium, Bacillus cereus, Staphylococcus aureus, Colletotrichum falcatum, Fusarium moniliforme, and Rhizoctonia solani. Among all extracts, ginger peel extracts exhibited maximum inhibition against all bacterial strains, while onion peel extracts exhibited zero inhibition against any bacterial strains. All extracts exhibited maximum inhibition against Staphylococcus aureus except onion peel extracts. Positive inhibition against all fungal strains was observed for all samples, with maximum inhibition against Colletotrichum falcatum. The outcomes of the study, therefore reveal that Agro-wastes have a powerful antimicrobial and antioxidant potency and thus can be used for many medicinal purposes, which will likewise be helpful in waste management and environmental safety.

Morpho-palynological study of Cyperaceae from wetlands of Azad Jammu and Kashmir using SEM and LM.

In this study 12 species of Cyperaceae have been studied for quantitative and qualitative observation of pollen grains through Light and scanning electron microscopy. Pollens of 12 species of Cyperaceae from different wetlands of Azad Jammu and Kashmir were collected. Morphological characters of pollen grains were then investigated under the Light and Scanning electron microscope. Two pollen types have been observed apolar and heteropolar. Shape of pollens was prolate (4 spp), sub-spheroidal (7 spp), and oblate (1 spp). Variation observed in exine sculpturing granular (4 spp), reticulate (1 spp), areolate-punctate (3 spp), and psilate (2 spp). Polar to equatorial ratio and fertility percentage of the pollens were also studied. Based on these micromorphlogical characters of pollens taxonomic keys have been made for the accurate identification of the members of Cyperaceae. The characteristics studied in present research work are very much valuable taxonomically and phytochemically for the identification of species of family Cyperaceae. Light microscope (LM) and Scanning electron microscope (SEM) were used for pollen observation, which play vital role in the taxonomical identification of species and provide sufficient information for taxonomist.

Extensively and pre-extensively drug resistant tuberculosis in clinical isolates of multi-drug resistant tuberculosis using classical second line drugs (levofloxacin and amikacin).

OBJECTIVE: To find out the frequency of Extensively Drug Resistant (XDR) and pre-XDR tuberculosis in clinical isolates of Multi-Drug Resistant (MDR) Tuberculosis (TB) by determining the susceptibilities against Levofloxacin and Amikacin (classical second line antituberculosis drugs). STUDY DESIGN: Adescriptive cross-sectional study. PLACE AND DURATION OF STUDY: Microbiology Department, Armed Forces Institute of Pathology (AFIP), Rawalpindi, from September 2011 to August 2013. METHODOLOGY: Amikacin (AK) and Levofloxacin (LEVO) were obtained in chemically pure form from Sigma (Taufkirchen, Germany). The breakpoint concentration used for AK was 1.0 µg/ml and for LEVO 2.0 µg/ml. Mycobacterial Growth Indicator Tube (MGIT) 960 system was used to carry out drug susceptibility testing as per recommended protocol. RESULTS: A total of 3 MDR-TB isolates (3%) turned out to be XDR-TB based upon simultaneous resistance to injectable second line antituberculosis drug AK and one of the fluoro-quinolones (LEVO). A total of 24 MDR-TB isolates (24%) were found to be pre-XDR based upon resistance to LEVO alone. Treatment status record of patients with XDR and pre-XDRTB isolates revealed that majority of patients had received fluoroquinolones (FQs) during the course of treatment. CONCLUSION: XDR-TB has started to emerge in MDR-TB isolates in our set up. The worrying sign is the high frequency of pre-XDR tuberculosis. Urgent steps need to be taken to stem the tide of pre-XDR-TB in our population. It is thus recommended to develop facilities to carry out drug susceptibility testing to monitor the status of pre-XDR and XDR-TB in our population.

In vitro activity of fosfomycin tromethamine against extended spectrum beta-lactamase producing urinary tract bacteria.

OBJECTIVE: To determine the in vitro activity of Fosfomycin tromethamine against extended spectrum beta-lactamase producing uropathogens. STUDY DESIGN: Experimental study. PLACE AND DURATION OF STUDY: Department of Microbiology, Armed Forces Institute of Pathology, Rawalpindi, from October 2011 to October 2012. METHODOLOGY: A total of 381 culture positive ESBL producing isolates from 2400 urine samples submitted over a period of one year were included in this study. Identification of isolates was done by standard biochemical profile of the organisms. The antimicrobial susceptibility of culture positive isolates was performed by disk diffusion method as recommended by Clinical Laboratory Standard Institute guidelines (CLSI). RESULTS: The antimicrobial activity of Fosfomycin to various isolates revealed that 93% of E. coli, 64% Klebsiella spp. 50% Proteus spp. 75% Enterobacter cloacae, 100% Citrobacter freundii, 100% Burkholderia spp. 100% Serratia spp. and 50% Stenotrophomonas maltophilia were susceptible to this chemical compound. CONCLUSION: Fosfomycin showed excellent effectiveness to most of the common ESBL producing bacteria such as E. coli, Klebsiella and Proteus spp.

Susceptibility pattern of extended spectrum ß-lactamase producing isolates in various clinical specimens.

OBJECTIVE: To determine the susceptibility pattern of extended spectrum ß-lactamase (ESBL) producing Gram negative isolates from various clinical specimens. STUDY DESIGN: Descriptive study. PLACE AND DURATION OF STUDY: Microbiology Department, Armed Forces Institue of Pathology, Rawalpindi, from January 2008 to January 2009. METHODOLOGY: A total of 308 ESBL producing isolates from various clinical specimens sent to AFIP for culture and sensitivity were identified using standard microbiological techniques and tested for antimicrobial susceptibility. At the same time screening for ESBL production was also done. ESBL production was confirmed by combination disc synergy method. The susceptibility pattern of isolates was then recorded in frequency percentages. RESULTS: Out of the 308 ESBL producing isolates more than 99% were susceptible to carbapenems, 84% to tazobactam/ piperacillin, 81% to sulbactam/cefoperazone, 12% to fluoroquinolones, 13% to cotrimoxazole, 59% to amikacin and 18% to gentamicin. Among the urinary isolates 49% were susceptible to Nitrofurontoin and only 5% to Pipemidic acid. CONCLUSION: Antibiotic choices in case of ESBL producing isolates are limited and at present only carbapenems can be regarded as treatment of choice. As empirical agents, beta-lactam/beta lactamase inhibitor combinations should be used cautiously for serious infections. Fluoroquinolones showed very poor efficacy. Amikacin can be used alternatively in such cases. Nitrofurantoin is still a good oral agent for treating UTI.

Galinsosides A and B, bioactive flavanone glucosides from Galinsoga parviflora.

Galinsosides A (1) and B (2), new flavanone glucosides together with two known flavanones, 7,3',4'-trihydroxyflavanone (3) and 3,5,7,3',4'-pentahydroxyflavanone (4) have been isolated from an ethyl acetate- soluble fraction of Galinsoga parviflora. Their structures were assigned on the basis of spectral studies. Compound 1 showed significant antioxidant and urease inhibitory activity while compound 2 was moderately active. On the other hand, 2 showed inhibitory potential against alpha-glucosidase.