Advancements in CHO metabolomics: techniques, current state and evolving methodologies.

Background: Investigating the metabolic behaviour of different cellular phenotypes, i.e., good/bad grower and/or producer, in production culture is important to identify the key metabolite(s)/pathway(s) that regulate cell growth and/or recombinant protein production to improve the overall yield. Currently, LC-MS, GC-MS and NMR are the most used and advanced technologies for investigating the metabolome. Although contributed significantly in the domain, each technique has its own biasness towards specific metabolites or class of metabolites due to various reasons including variability in the concept of working, sample preparation, metabolite-extraction methods, metabolite identification tools, and databases. As a result, the application of appropriate analytical technique(s) is very critical. Purpose and scope: This review provides a state-of-the-art technological insights and overview of metabolic mechanisms involved in regulation of cell growth and/or recombinant protein production for improving yield from CHO cultures. Summary and conclusion: In this review, the advancements in CHO metabolomics over the last 10 years are traced based on a bibliometric analysis of previous publications and discussed. With the technical advancement in the domain of LC-MS, GC-MS and NMR, metabolites of glycolytic and nucleotide biosynthesis pathway (glucose, fructose, pyruvate and phenylalanine, threonine, tryptophan, arginine, valine, asparagine, and serine, etc.) were observed to be upregulated in exponential-phase thereby potentially associated with cell growth regulation, whereas metabolites/intermediates of TCA, oxidative phosphorylation (aspartate, glutamate, succinate, malate, fumarate and citrate), intracellular NAD+/NADH ratio, and glutathione metabolic pathways were observed to be upregulated in stationary-phase and hence potentially associated with increased cell-specific productivity in CHO bioprocess. Moreover, each of technique has its own bias towards metabolite identification, indicating their complementarity, along with a number of critical gaps in the CHO metabolomics pipeline and hence first time discussed here to identify their potential remedies. This knowledge may help in future study designs to improve the metabolomic coverage facilitating identification of the metabolites/pathways which might get missed otherwise and explore the full potential of metabolomics for improving the CHO bioprocess performances.

Fabrication of mulberry leaf extract (MLE)- and tasar pupal oil (TPO)-loaded silk fibroin (SF) hydrogels and their antimicrobial properties.

Biocomposites have gained tremendous advantages over synthetic composites due to their biocompatibility, sustainable degradation, and ability to easily combine with other substances. In the present study, we have prepared silk fibroin (SF) hydrogel, mulberry leaf extract (MLE), tasar pupal oil (TPO), and their composites, such as TPO-loaded SF hydrogel and MLE-loaded SF hydrogel, and characterized them by using a phase contrast microscope (PCM), scanning electron microscope (SEM) SEM- EDX, and Fourier transform infrared spectroscopy (FTIR). In addition, 1H-NMR was used for profiling of mulberry leaf extract and GC-MS was used to find tasar pupal oil composition. Further, the disc diffusion method evaluated their antimicrobial activities against S. aureus, E. coli, A. flavus, and A. brassicae. PCM, SEM, and FTIR results validated the conjugation of MLE and SF hydrogel composite; 1H-NMR confirmed the 41 metabolites in MLE, and GC-MS established the composition of tasar pupal oil. Since both composites, such as TPO-loaded SF hydrogel and MLE-loaded SF hydrogel, reduced the S. aureus and E. coli activities at all tested concentrations, the antibacterial results were unambiguous in their conclusion. S. aureus could only be inhibited by SF hydrogel at a high concentration (300 g/ml), despite suppressing E. coli growth at all tested concentrations. At 300 g/ml, MLE demonstrated antibacterial action against S. aureus. Furthermore, at a dosage of 300 g/ml, TPO inhibited both S. aureus and E. coli. Both mulberry leaf extract (at 200 and 300 g/ml) and the MLE-loaded SF hydrogel composite displayed antifungal activity against A. flavus at all tested concentrations (100, 200, and 300 g/ml).

Prospective Study to Evaluate the Role of Multidetector Computed Tomography in Evaluation of Paranasal Sinus Pathologies.

BACKGROUND: The use of computed tomography (CT) combined with functional endoscopic sinus surgery (FESS) has empowered the modern sinus surgeon to treat patients more effectively, facilitating reduced morbidity and complications. This study aimed to evaluate the role of multidetector computed tomography (MDCT) in the evaluation of paranasal sinus pathologies. MATERIALS & METHODS: This cross-sectional study was conducted among the adult subjects attending the department of radiology at a tertiary care center in Lucknow for CT scan of paranasal sinuses for suspected paranasal sinus pathology from August 2018 to January 2020. The study included subjects above 12 years of age visiting the facility for CT of paranasal sinuses with a suspected paranasal sinus pathology and were also undergoing FESS. RESULTS: A total of 74 patients falling in the sampling frame were enrolled in the study. Majority of cases were aged <40 years (n=41; 55.4%). Age group of 21-30 years was most affected (24.3%). Mean age of patients was 38.39±14.48 years. Inflammatory pathologies diagnosed on MDCT-included sinusitis (n=25), sinonasal polyps (n=17), sinusitis with polyps (n=15), and mucocele (n=2), respectively. FESS/histopathological diagnosis was done in 73 cases and revealed inflammatory pathology in 59/73 (80.8%) and neoplastic pathology in 14 (19.2%) cases. FESS/histopathological break up of inflammatory pathologies included 22 cases of sinusitis, 20 cases of sinonasal polyps, 14 cases of sinusitis with polyps, and three cases of mucocele. Agreement between MDCT and final diagnosis was seen in 67/74 (90.5%) cases assessed. CONCLUSION: The findings of the present study showed that MDCT is a useful modality for preoperative assessment of paranasal sinuses. With its high precision in diagnosis, it can help in further treatment planning and management in patients with paranasal sinus pathologies.

Plant isoquinoline alkaloids: Advances in the chemistry and biology of berberine.

Alkaloids are one of the most important classes of plant bioactives. Among these isoquinoline alkaloids possess varied structures and exhibit numerous biological activities. Basically these are biosynthetically produced via phenylpropanoid pathway. However, occasionally some mixed pathways may also occur to provide structural divergence. Among the various biological activities anticancer, antidiabetic, antiinflammatory, and antimicrobial are important. A few notable bioactive isoquinoline alkaloids are antidiabetic berberine, anti-tussive codeine, analgesic morphine, and muscle relaxant papaverine etc. Berberine is one of the most discussed bioactives from this class possessing broad-spectrum pharmacological activities. Present review aims at recent updates of isoquinoline alkaloids with major emphasis on berberine, its detailed chemistry, important biological activities, structure activity relationship and implementation in future research.

Fluorinated benzylidene indanone exhibits antiproliferative activity through modulation of microtubule dynamics and antiangiogenic activity.

The application of fluorine in drug design has been understood significantly by the medicinal chemists in recent years. Modulation of tubulin-microtubule dynamics is one of the most effective targets for cancer chemotherapeutics. A logically designed and identified lead compound, fluorinated benzylidene indanone 1 has been extensively evaluated for cancer pharmacology. It occupied colchicine binding pocket acting as microtubule destabilizer and induced a G2/M phase arrest in MCF-7 cells. Compound 1 exerted an antiangiogenic effect in MCF-7 cells by down-regulating Vascular Endothelial Growth Factor (VEGF) and Hypoxia Inducible Factor-α (HIF-α). In in-vivo efficacy in C3H/Jax mice mammary carcinoma model, benzylidene indanone 1 reduced tumour volumes by 48.2%. Further in acute oral toxicity studies compound 1 was well tolerated and safe up to 1000 mg/kg dose in Swiss albino mice. The fluorinated benzylidene indanone 1, a new chemical entity (NCE) can further be optimized for better efficacy against breast adenocarcinoma.1.

Posterior capsule rupture with herniation of lens fragment following blunt ocular trauma.

Posterior capsule rupture with herniated lens fragment in the vitreous cavity on magnetic resonance imaging has not been reported in India until now; however, it has been reported in other countries. Therefore, this study reports the case of a 15-year-old boy presenting with posttraumatic loss of vision in the right eye due to posterior capsular rupture and herniation of lens material into the vitreous cavity, which was detected by B-scan ultrasonography and magnetic resonance imaging as no ophthalmic examination was possible due to the posttraumatic cataract. The patient was treated by lens aspiration with anterior chamber vitrectomy and placement of posterior chamber intraocular lens, with the patient achieving 6/6 visual acuity postoperatively. This case is unusual due to the rarity of the findings, and it highlights the crucial role of imaging in achieving timely diagnosis and surgery to restore vision in the affected eye.