Growth Medium Research Articles

C1s targeting antibodies inhibit the growth of cutaneous squamous carcinoma cells

Cutaneous squamous cell carcinoma (cSCC) is the most common metastatic skin cancer. The incidence of cSCC is increasing globally and the prognosis of metastatic disease is poor. Currently there are no specific targeted therapies for advanced or metastatic cSCC. We have previously shown abundant expression of the complement classical pathway C1 complex components, serine proteases C1r and C1s in tumor cells in invasive cSCCs in vivo, whereas the expression of C1r and C1s was lower in cSCCs in situ, actinic keratoses and in normal skin. We have also shown that knockdown of C1s expression results in decreased viability and growth of cSCC cells by promoting apoptosis both in culture and in vivo. Here, we have studied the effect of specific IgG2a mouse monoclonal antibodies TNT003 and TNT005 targeting human C1s in five primary non-metastatic and three metastatic cSCC cell lines that show intracellular expression of C1s and secretion of C1s into the cell culture media. Treatment of cSCC cells with TNT003 and TNT005 significantly inhibited their growth and viability and promoted apoptosis of cSCC cells. These data indicate that TNT003 and TNT005 inhibit cSCC cell growth in culture and warrant further investigation of C1s targeted inhibition in additional in vitro and in vivo models of cSCC.

Labeling of Cell Surface Proteins at the Drosophila Larval Neuromuscular Junction Using Binding Partner Peptides.

Determining the precise localization of interacting proteins provides fundamental insight into their putative function. Classically, immunolabeling of endogenous proteins or generating tagged versions of proteins has been used to localize interacting proteins. However, in many cases, the interacting partner of a protein of interest is unknown. For cell surface proteins, it is possible to determine the localization of interacting proteins if one of the binding partners is known. This approach is based on generating purified, recombinant, tagged extracellular domains (ECDs) of a protein of interest, and incubating tissue to allow the recombinant protein to bind to its interacting partner(s). In this protocol, we detail the cloning of secreted, tagged ECDs from cell surface proteins, transfection of cloned plasmids into S2 cells, collection of secreted domains, concentration of the cell culture medium to enrich for the ECDs, and labeling of tissue with these ECDs.

Valorisation of Spent Yeast Fermentation Media through Compositional-Analysis-Directed Supplementation

Spent fermentation media from bioprocessing represent a significant waste stream, and interest in recycling them as part of the developing circular bioeconomy is growing. The potential to reuse yeast spent culture media (YSM) to feed secondary bacterial fermentations producing recombinant protein was investigated in this study. Elemental and amino acid compositional analysis using inductively coupled plasma mass spectrometry (ICP-MS) and LC-MS/MS identified significant differences in the concentrations of 6 elements and 18/20 amino acids in YSM compared with rich microbiological media (LB). Restoration of levels of magnesium and sodium through addition of their salts and amino acids from tryptone supplementation led to the expression of equivalent titres of recombinant proteins by E. coli (0.275 g/L), compared to that in LB media (0.296 g/L) and BMMY media (0.294 g/L) in shake flask culture. When this supplementation strategy was employed in a bioreactor system, we observed a significant increase in recombinant protein titre using the supplemented YSM (2.29 (±0.02) g/L) over that produced using LB media (1.29 (±0.09) g/L). This study demonstrates through highly sensitive compositional analysis and identification of supplementation strategies the potential to valorise spent media from yeast fermentations that underpin industrial processes of significant scale, creating a circular approach to waste stream management.

Assessing the In Vitro Individual and Combined Effect of Arthrobotrys oligospora and A. musiformis (Orbiliales) Liquid Culture Filtrates against Infective Larvae of the Sheep Blood-Feeding Nematode Haemonchus contortus (Trichostrongylidae).

Arthrobotrys species are nematophagous fungi that secrete extracellular nematocidal products (ECP). The individual and combined effects of ECP from Arthrobotrys oligospora (Ao) and A. musiformis (Am) growth in liquid media against Haemonchus contortus L3 (HcL3) were assessed. The isolation, morphological (MI) and molecular identification (Mol-I), assessment of nematocidal activity (NA) of fungal liquid culture filtrates (LCF) in two liquid media alone and in combination and the myco-compound profile identification (MCP) were performed. The MI suggested that the fungi corresponded to the species Ao and Am. This result was confirmed by PCR analysis followed by sequencing, alignment and a phylogenetic analysis. Likewise, the highest Hc mortalities were 91.4% with individual LCF of Am and 86.2% with those of Ao at the highest concentration (100 mg/mL) in Czapek-Dox Broth. The combination of both LCF resulted in a similarly high larval mortality with no statistical differences in relation to individual activity (p > 0.05). The MCP showed the presence of alkaloids in both fungi. Coumarins, sterols and saponins were found only in Ao. Both fungi produced ECP with a high NA that could be identified and assessed in future studies as potential natural anthelmintic compounds.

Development of a Quorum Sensing-Mediated Bacterial Autolytic System in Escherichia coli for Automatic Release of Intracellular Products.

Escherichia coli, one of the most efficient expression hosts for recombinant proteins, is widely used in chemical, medical, food, and other industries. De novo engineering of gene regulation circuits and cell density-controlled E. coli cell lysis are promising directions for the release of intracellular bioproducts. Here, we developed an E. coli autolytic system, named the quorum sensing-mediated bacterial autolytic (QS-BA) system, by incorporating an acyl-homoserine lactone (AHL)-based YasI/YasR-type quorum sensing circuit from Pseudoalteromonas into E. coli cells. The results showed that the E. coli QS-BA system can release the intracellular bioproducts into the cell culture medium in terms of E. coli cell density, which offers an environmentally-friendly, economical, efficient, and flexible E. coli lysis platform for production of recombinant proteins. The QS-BA system has the potential to serve as an integrated system for the large-scale production of target products in E. coli for medical and industrial applications.

Hypoxia-induced activation of HIF-1alpha/IL-1beta axis in microglia promotes glioma progression via NF-κB-mediated upregulation of heparanase expression

BackgroundGlioma is a common tumor that occurs in the brain and spinal cord. Hypoxia is a crucial feature of the tumor microenvironment. Tumor-associated macrophages/microglia play a crucial role in the advancement of glioma. This study aims to illuminate the detailed mechanisms by which hypoxia regulates microglia and, consequently, influences the progression of glioma.MethodsThe glioma cell viability and proliferation were analyzed by cell counting kit-8 assay and 5-ethynyl-2’-deoxyuridine assay. Wound healing assay and transwell assay were implemented to detect glioma cell migration and invasion, respectively. Enzyme-linked immunosorbent assay was conducted to detect protein levels in cell culture medium. The protein levels in glioma cells and tumor tissues were evaluated using western blot analysis. The histological morphology of tumor tissue was determined by hematoxylin-eosin staining. The protein expression in tumor tissues was determined using immunohistochemistry. Human glioma xenograft in nude mice was employed to test the influence of hypoxic microglia-derived interleukin-1beta (IL-1β) and heparanase (HPSE) on glioma growth in vivo.ResultsHypoxic HMC3 cells promoted proliferation, migration, and invasion abilities of U251 and U87 cells by secreting IL-1β, which was upregulated by hypoxia-induced activation of hypoxia inducible factor-1alpha (HIF-1α). Besides, IL-1β from HMC3 cells promoted glioma progression and caused activation of nuclear factor-κB (NF-κB) and upregulation of HPSE in vivo. We also confirmed that IL-1β facilitated HPSE expression in U251 and U87 cells by activating NF-κB. Hypoxic HMC3 cells-secreted IL-1β facilitated the proliferation, migration, and invasion of U251 and U87 cells via NF-κB-mediated upregulation of HPSE expression. Finally, we revealed that silencing HPSE curbed the proliferation and metastasis of glioma in mice.ConclusionHypoxia-induced activation of HIF-1α/IL-1β axis in microglia promoted glioma progression via NF-κB-mediated upregulation of HPSE expression.

Optimized LC-MS/MS methods for quantifying antibody–drug conjugate payloads in cell culture media containing phenol red

Optimized LC-MS/MS methods for quantifying antibody–drug conjugate payloads in cell culture media containing phenol red

Response surface methodology mediated optimization of phytosulfokine and plant growth regulators for enhanced protoplast division, callus induction, and somatic embryogenesis in Angelica Gigas Nakai

BackgroundAngelica Gigas (Purple parsnip) is an important medicinal plant that is cultivated and utilized in Korea, Japan, and China. It contains bioactive substances especially coumarins with anti-inflammatory, anti-platelet aggregation, anti-cancer, anti-diabetic, antimicrobial, anti-obesity, anti-oxidant, immunomodulatory, and neuroprotective properties. This medicinal crop can be genetically improved, and the metabolites can be obtained by embryonic stem cells. In this context, we established the protoplast-to-plant regeneration methodology in Angelica gigas.ResultsIn the present investigation, we isolated the protoplast from the embryogenic callus by applying methods that we have developed earlier and established protoplast cultures using Murashige and Skoog (MS) liquid medium and by embedding the protoplast in thin alginate layer (TAL) methods. We supplemented the culture medium with growth regulators namely 2,4-dichlorophenoxyaceticacid (2,4-D, 0, 0.75, 1.5 mg L− 1), kinetin (KN, 0, 0.5, and 1.0 mg L− 1) and phytosulfokine (PSK, 0, 50, 100 nM) to induce protoplast division, microcolony formation, and embryogenic callus regeneration. We applied central composite design (CCD) and response surface methodology (RSM) for the optimization of 2,4-D, KN, and PSK levels during protoplast division, micro-callus formation, and induction of embryogenic callus stages. The results revealed that 0.04 mg L− 1 2,4-D + 0.5 mg L− 1 KN + 2 nM PSK, 0.5 mg L− 1 2,4-D + 0.9 mg L− 1 KN and 90 nM PSK, and 1.5 mg L− 1 2,4-D and 1 mg L− 1 KN were optimum for protoplast division, micro-callus formation and induction embryogenic callus. MS basal semi-solid medium without growth regulators was good for the development of embryos and plant regeneration.ConclusionsThis study demonstrated successful protoplast culture, protoplast division, micro-callus formation, induction embryogenic callus, somatic embryogenesis, and plant regeneration in A. gigas. The methodologies developed here are quite useful for the genetic improvement of this important medicinal plant.

Alternative Media from Mangrove Fruit for the Growth of Shigella dysentery, Escherichia coli, and Salmonella typhi bacteria

Objectives: Mangroves are rich in bioactive substances that support bacterial growth. The fruit of the mangrove tree can serve as a valuable source of carbohydrates, making it a potential substitute for Nutrient Agar (NA) as a growth medium. Methodology and Results: The samples in this study were suspensions of Shigella dysentery, Escherichia coli, and Salmonella typhiwith 16 replications. Thus, the total number of samples in this study was 94 samples. The samples were isolated and identified as colony growth on alternative Mangrove and NA media. The results showed that in alternative media prepared from Mangrove fruit flour, there was a significant growth difference (p-value <0,01) between S. dysentery bacteria with an average of 61 colonies, E. coli bacteria with an average of 2 colonies, and S. typhi bacteria with an average of 2 colonies. The comparison between media of Mangrove and NA also showed a significant growth difference with p-value < 0,01. Conclusions: Mangrove media has been found to be less conducive to the growth of S. dysentery, E. coli, and S. typhi bacteria, despite being a more cost-effective option compared to NA. Notably, S. dysenteriae bacteria exhibit greater potential for propagation in this medium. Keywords: Alternative, culture, mangrove, media, nutrient agar

Optimizing conditions of mycelial inoculum immobilized in Ca-alginate beads: a case study in ectomycorrhizal fungus Astraeus odoratus.

Ectomycorrhizal inoculum has emerged as a critical tool for forest restoration, especially under challenging climate change conditions. The inoculation of selective ectomycorrhizal fungi can enhance seedling survival and subsequent growth in the field. This study optimized the liquid media for mycelial growth of Astraeus odoratus strain K1 and the sodium alginate solution composition for enhanced mycelial viability after entrapment. Using Modified Melin-Norkrans as the optimal media for mycelial cultivation and 2% sodium alginate supplemented with Czapek medium, 0.25% activated charcoal, 5% sucrose, and 5% sorbitol in the alginate solution yielded the highest viability of A. odoratus mycelia. Preservation in distilled water and 10% glycerol at 25°C for 60 days proved to be the most effective storage condition for the alginate beads. Both fresh and preserved alginate beads were tested for colonizing on Hopea odorata Roxb. seedlings, showing successful colonization and ectomycorrhizal root formation, with over 49% colonization. This study fills a crucial gap in biotechnology and ectomycorrhizal inoculum, paving the way for more effective and sustainable forest restoration practices.

Oil-in-Water Emulsions Probed Using Fluorescence Multivariate-Curve-Resolution Spectroscopy.

Hydrophobic surfaces in contact with aqueous media are omnipresent in nature. A plethora of key biological and physiological processes occur at the interface of immiscible fluids. Besides its fundamental importance, probing such interfaces is rather challenging, especially when one medium is bathed in the other. Herein, we demonstrate a fluorescence-based method that probes the oil-water interface and interfacial processes through surface dielectric perturbations. The fluorescence response of Nile Red is measured in hexadecane in water nanoemulsions. Three major spectral components appear: two from the bulk liquid media (hexadecane and water) and a distinct band at around 640 nm due to the interfacial component. Such spectra are deconvoluted using the multivariate-curve-resolution algorithm, and interface-correlated fluorescence spectra are attained. The influence of anionic sodium dodecylbenzenesulfonate (SDBS) and cationic cetyltrimethylammonium bromide (CTAB) surfactants on the oil-water interface is elucidated with concentration-dependent measurements. A charge-dependent spectral shift is observed. The interface correlated band at 641 nm for bare hexadecane nanoemulsions red shifts in the presence of anionic surfactants, indicating an apparent dielectric increase. In contrast, the same band gradually blue shifts with increasing cationic surfactant concentration, indicating an apparent interface dielectric decrease. Such a method can be utilized to probe alterations at interfaces beyond the oil/water interface.

Utilization of organic-residues as potting media: Physico-chemical characteristics and their influence on vegetable production.

Soilless agriculture is acknowledged worldwide because it uses organic leftovers as a means of supporting intensive and efficient plant production. However, the quality of potting media deteriorates because of lower nutrient content and excessive shrinkage of most organic materials. A current study was undertaken to identify the optimal blend of locally available organic materials with desirable qualities for use as potting media. Therefore, different ingredients, viz., Pinus roxburghii needles, sugarcane bagasse, and farmyard manure were used alone or in combination as potting media to test their suitability by growing spinach as a test crop. Results showed that an increase in Pinus roxburghii needles and sugarcane bagasse decreased medium pH and electrical conductivity. Higher pH and electrical conductivity were recorded for the treatments having a higher farmyard manure ratio (≥50%) in combination. Except for pine needles 100%, pH and electrical conductivity were in the recommended range. The growth attributes include, leaves plant-1, shoot length, fresh- and dry shoot weight along with plant macronutrients (nitrogen, phosphorous, and potassium) and micronutrients (iron, copper, manganese, and zinc) content were higher in treatment pine needles 50%+farmyard manure 50% followed by pine needles 25%+farmyard manure 50%+sugarcane bagasse 25%. Moreover, the particular treatment of pine needles 50%+farmyard manure 50% exhibited the highest concentrations of macro- (nitrogen, phosphorus, and potassium) as well as micronutrients (iron, copper, manganese, and zinc) in the potting media following the harvest. This study highlights the potential of utilizing agro-industrial litter/waste as a soilless growing medium for spinach production under greenhouse conditions. When employed in appropriate proportions, this approach not only addresses disposal concerns but also proves effective for sustainable cultivation. Further research is needed to investigate the use of these wastes as potting media by mixing various particle-size ingredients.

Platelet-rich fibrin (PRF) modified nano-hydroxyapatite/chitosan/gelatin/alginate scaffolds increase adhesion and viability of human dental pulp stem cells (DPSC) and osteoblasts derived from DPSC

Bone tissue regeneration strategies have incorporated the use of natural polymers, such as hydroxyapatite (nHA), chitosan (CH), gelatin (GEL), or alginate (ALG). Additionally, platelet concentrates, such as platelet-rich fibrin (PRF) have been suggested to improve scaffold biocompatibility. This study aimed to develop scaffolds composed of nHA, GEL, and CH, with or without ALG and lyophilized PRF, to evaluate the scaffold's properties, growth factor release, and dental pulp stem cells (DPSC), and osteoblast (OB) derived from DPSC viability. Four scaffold variations were synthesized and lyophilized. Then, degradation, swelling profiles, and morphological analysis were performed. Furthermore, PDGF-BB and FGF-B growth factors release were quantified by ELISA, and cytotoxicity and cell viability were evaluated. The swelling and degradation profiles were similar in all scaffolds, with pore sizes ranging between 100 and 250 μm. FGF-B and PDGF-BB release was evidenced after 24 h of scaffold immersion in cell culture medium. DPSC and OB-DPSC viability was notably increased in PRF-supplemented scaffolds. The nHA-CH-GEL-PRF scaffold demonstrated optimal physical-biological characteristics for stimulating DPSC and OB-DPSC cell viability. These results suggest lyophilized PRF improves scaffold biocompatibility for bone tissue regeneration purposes.

БИОТЕХНОЛОГИЧЕСКИЙ ПОТЕНЦИАЛ НОВЫХ ШТАММОВ УГЛЕВОДОРОДОКИСЛЯЮЩИХ БАКТЕРИЙ

The ingress of oil has a depressing effect on the soil biocenosis, which is enhanced in the presence of other pollutants (heavy metals, chlorides, herbicides), which ultimately leads to the withdrawal of significant land areas (including agricultural ones) from the target turnover. For the bioremediation of such areas, oil-degrading microorganisms are needed that are resistant to additional pollutants, as well as a set of useful properties that increase the efficiency of cleaning and restoring disturbed soils. In this work, three isolates were isolated that actively grew in a liquid medium containing oil. As a result of sequencing the nucleotide sequence of the 16S rRNA gene, it was reliably established that all strains belong to the species Acinetobacter calcoaceticus . The bacteria showed a high oil-oxidizing ability (89.7-97.0%), which was assessed by the degree of destruction of the aliphatic fraction, including in the presence of herbicides based on 2,4-dichlorophenoxyacetic acid (Octapon extra), imazethapyr (Tapir) and tribenuron-methyl ( Special forces) (degree of biodestruction 40.4-54.1%). Microorganisms used aromatic hydrocarbons (including polycyclic ones) as a carbon source. They demonstrated high cell surface hydrophobicity (78-85%) with respect to hexadecane and exhibited emulsifying activity of more than 50%. The bacteria were tolerant to the herbicides Tapir and Spetsnaz in amounts up to 1% volume (mass). The herbicide Octapon extra was more toxic to bacteria - the growth of all strains occurred at its concentration in the medium not higher than 0.5% volume. Microorganisms showed resistance to sodium chloride in an amount of 3.0-5.0% and lead ions (1.00-1.25 g/l). They produced the enzyme lipase and were capable of fixing atmospheric nitrogen and dissolving inorganic phosphate (including in the presence of oil or herbicides). The studied Acinetobacter spp. stimulated the growth of barley (aboveground and underground parts), especially the A. calcoaceticus strain P32 (elongation of shoots and roots by 15.3 and 49.5%, respectively). The data obtained indicate that all three strains have certain prospects for use in the purification of complexly contaminated soils.

Controlling the growth of uncapped silver nanoparticles on poly methacrylic acid nanospheres: The effect of polymer’s free surface on the antimicrobial properties

In this study, the control over the growth of uncapped silver nanoparticles on the surface of poly methacrylic acid nanospheres is presented. The pH of the growth medium and the size of the polymer nanospheres were studied as parameters in relation to the antimicrobial activity of the resulting structures against Escherichia coli and Staphylococcus aureus. Adjustment of the growth medium pH has a direct effect on the number of silver nanoparticles grown on the surface of the nanospheres, while the size of the nanospheres affects the resulting silver nanoparticle size distribution. The minimum bactericidal concentration was determined to 8 and 32 μg/mL against E. coli and S. aureus respectively, whereas the antimicrobial efficiency of the resulting nanospheres was enhanced in structures with increased free surface. Hence, the contribution of the as synthesized structures to the antimicrobial activity is attributed to the polymer’s ability to form stable suspensions in the aqueous bacterial growth medium, thus promoting the dissolution of silver nanoparticles to Ag cations with antimicrobial activity.

Bioremediation of Liquid Waste of Olive Presses (Al-Jeft water) by Using the Biomass of the Local Isolate of The Fungus Helvella bachu

The study included investigating the potential of the biomass of the local isolate of the fungus Helvella bachu in bioremediation of the liquid waste of olive presses (Jeft water) at two concentrations of 10%-20%. The mycelium was inoculated on solid media treated with olive press water, with concentrations of 10% and 20%, for several incubation days 2, 4, and 6, respectively. We observed that the diameter of the fungal colony is directly proportional to the increase in the incubation period, as the largest diameter of the fungal colony was 13.56 mm and 11.46 mm after 6 days of incubation at a concentration of 10% and 20% of Jeft water, respectively, compared to the diameter of the fungal colony growing in untreated medium, which it reached 15 mm, while the least diameter of the fungal colony was 6.00 and 5.36 mm at a concentration of 10% and 20% of Jeft water, respectively, after two days of incubation compared to the control sample. As for using liquid media, the results revealed a decrease in the total phenol concentration, reaching 2.14 mg / 100 g after 30 days of treatment with a concentration of 20% of Jeft water, compared to the control treatment of 23.58 mg / 100. The percentage of black color removal was 64.39% at a concentration of 10% and an incubation period of 30 days, while at a concentration of 20%, the percentage of color removal was 49.53% and 42.10% after 30 days of incubation. The biomass of the fungus isolate was able to reduce the chemical oxygen demand in both concentrations of Jeft water used with different responses. The percentage of chemical oxygen requirement was 65.83% at a concentration of 10% of Jeft water after 30 days of treatment, while the percentage of COD was 8.08% at a concentration of 20% for the same incubation period above.

Experimental study of the antibacterial effect of anodic dissolution of a copper electrode used in endodontic treatment of teeth

BACKGROUND: One of the possible ways to improve the quality of dental treatment with obliterated root canals is the use of transcanal direct current exposure. AIM: The aim of the study was to investigate the comparative antibacterial activity of anodic dissolution of copper and silver-copper electrodes used in the endodontic treatment of teeth in the experiment. MATERIALS AND METHODS: An experimental study was performed by implementing a technique for the automatic cultivation of microorganisms in liquid culture media. We used for the study clinical isolates of individual strains of bacteria and yeasts, namely: S. constellatus, P. intermedia, C. albicans, as well as mixed cultures: 1) S. constellatus + F. nucleatum; 2) Streptococcus sanguis + Enterococcus faecium obtained from the root canals of the teeth during treatment of chronic pulpitis. RESULTS: The results of the study showed that anodic dissolution of both silver-copper and copper electrodes had a significant and, in general, unidirectional antibacterial effect. At the same time, it was found that, if for clinical isolate of P. intermedia the use of a silver-copper electrode was more effective, then for S. constellatus and C. Albicans strains, as well as for mixed cultures of pathogenic microorganisms S. constellatus + F. nucleatum and Streptococcus sanguis + Enterococcus faecium anodic dissolution of a copper electrode showed a more pronounced antibacterial effect. CONCLUSION: In endodontic treatment of teeth with partially obliterated root canals, along with anodic dissolution of silver-copper electrodes, it is possible to use anodic dissolution of copper electrodes as a means capable of having a pronounced antibacterial effect.

Isolation of Potential Azo dye Degrading/Tolerating Bacteria from Polluted Environment

Various industries including the food and textile industries result in the release of azo dyes into the environment. These dyes are known to exert toxic effects on humans, animal and plants. Biological methods of azo dye bioremediation is a solution for the detoxification of azo dye in the environment. The present study was undertaken to isolate bacteria with potential for azo dye bioremediation. From five polluted environment samples, eight bacteria were isolated in azo dye supplemented Nutrient agar. Six isolates (75%, n=8) decolorized azo dye completely following 24 hours of incubation. The control tube with no bacterial inoculation remained blue, indicating that only microbial biotransformation processes were taking place. In each case, a blue-colored ring remained at the top, indicating the anoxic nature of the dye decolorization process. All isolates grew in presence of 400 ppm, 60% tolerated 600 ppm and 20% tolerated 800 ppm, whereas 1000ppm was inhibitory for growth of all isolates. Dead autoclaved cells of three representative isolates were tested for biosorption potential. Only one isolate turned the methylene blue supplemented nutrient broth colorless. This explains that this isolate was not metabolizing methylene blue rather it bound the dye to the cell structure. Two isolates did not show biosorption abilities indicating that their mechanism of bioremediation was enzymatic reduction. 16s rDNA sequencing identified two of the isolates as Lysinibacillus capsici and Stenotrophomonas muris. Bangladesh J Microbiol, Volume 40, Number 2, December 2023, pp 56-59

Unveiling the Significance of Shodhan with Special Emphasis on Samanya Shodhana of Pittala

Shodhana, a crucial procedure in the pharmaceutical preparation of metals and minerals, aims to eliminate impurities and undesirable properties from raw materials, rendering them suitable for further processing. This study focuses on Samanya Shodhana of Pittala, a Mishra Loha, following the guidelines outlined in Rasa Ratna Samucchya. The process involves heating raw Pittala until red-hot and subsequently quenching it in Tila Taila, Takra, Gomutra, Kanji, and Kulatha Kwatha for seven iterations. Post-Shodhana, observations were made regarding the weight loss of Pittala, as well as pH and color changes in the respective liquid media. Results indicate a total weight loss of 7.0% in Pittala after Samanya Shodhana, suggesting effective impurity eradication.

Characterization of bacteriophages infecting multidrug-resistant uropathogenic Escherichia coli strains

Uropathogenic Escherichia coli (UPEC) is the most common causative agent of urinary tract infections, and strains that are resistant to antibiotics are a major problem in treating these infections. Phage therapy is a promising alternative approach that can be used to treat infections caused by polyresistant bacterial strains. In the present study, 16 bacteriophages isolated from sewage and surface water were investigated. Phage host specificity was tested on a collection of 77 UPEC strains. The phages infected 2–44 strains, and 80% of the strains were infected by at least one phage. The susceptible E. coli strains belonged predominantly to the B2 phylogenetic group, including strains of two clones, CC131 and CC73, that have a worldwide distribution. All of the phages belonged to class Caudoviricetes and were identified as members of the families Straboviridae, Autographiviridae, and Drexlerviridae and the genera Kagunavirus, Justusliebigvirus, and Murrayvirus. A phage cocktail composed of six phages – four members of the family Straboviridae and two members of the family Autographiviridae – was prepared, and its antibacterial activity was tested in liquid medium. Complete suppression of bacterial growth was observed after 5–22 hours of cultivation, followed by partial regrowth. At 24 hours postinfection, the cocktail suppressed bacterial growth to 43–92% of control values. Similar results were obtained when testing the activity of the phage cocktail in LB and in artificial urine medium. The results indicate that our phage cocktail has potential to inhibit bacterial growth during infection, and they will therefore be preserved in the national phage bank, serving as valuable resources for therapeutic applications.