A Comprehensive Computed Tomographic Analysis of Pneumatization Pattern of Sphenoid Sinus and Their Association with Protrusion/Dehiscence of Vital Neurovascular Structure in a Pakistani Subgroup.

AIM: To determine the clinically significant association between pneumatization types of the sphenoid sinus (SS) and protrusion/ dehiscence of the optic nerve (ON) and the internal carotid artery (ICA). MATERIAL AND METHODS: This prospective cross-sectional study was conducted between November 2020 and April 2021 at the Dow Institute of Radiology, Dow University of Health Sciences, Karachi. This study examined 300 computed tomography (CT) PNS patients aged 18-60 years. The forms of SS pneumatization, extent of pneumatization to the greater wing (GW), anterior clinoid process (ACP), and pterygoid process (PP), as well as the protrusion/dehiscence of the ON and ICA were examined. A statistical relationship was identified between pneumatization type and protrusion/dehiscence of the ON and ICA. RESULTS: The study included 171 men and 129 women with an average age of 39.28 ± 10.9 years. The most commonly encountered pneumatization type was postsellar (63.3%), followed by sellar (27.3%), presellar (8.7%), and conchal (0.75%). The most frequent extended pneumatization was observed up to PP (44%), followed by ACP (31.33%), and GW (16.67%). The rate of dehiscence of the ON and ICA was less than that of protrusion of the same structures. The association between postsellar and sellar pneumatization types and protrusion of the ON and ICA was statistically significant (p < 0.001), with the postsellar type showing more protrusions of the ON and ICA than the sellar type. CONCLUSION: The pneumatization type of SS has a significant impact on the protrusion/dehiscence of adjacent vital neurovascular structures and should be mentioned in CT reports to alert surgeons for any disastrous intraoperative complications and outcomes.

Differentiation of CD117+ Amniotic Fluid Stem Cells towards Nephron Progenitors.

Objective: The study aimed at isolation of CD117+ stem cells from amniotic fluid samples followed by their invitro differentiation towards nephron progenitors that can be potentially used for regenerative medicine studies and to further understand pathways involved in renal pathogenesis. Methods: This experimental study was conducted at Dow Research Institute of Biotechnology and Biomedical Sciences (DRIBBS), Dow University of Health Sciences, OJHA Campus Karachi from November 2019 to December 2020. After patient consent, a Pfannenstiel incision was performed by the gynecologist through abdominal and uterine muscles without cutting into Amniotic Membrane. Using a needle of 5CC syringe connected to sterile Redivac bottle, a blunt end insertion was passed through the membrane and the amniotic fluid was aseptically sucked into Redivac bottle, the ice bag was used for transporting amniotic fluid from hospital to the lab and samples were processed within 60 minutes after collection. Amniotic fluid was centrifuged at 4º C for 20 minutes at 1400xg. After centrifugation the cell pellet was treated for analysis of CD117+ cells using flowcytometry, once small percentage of CD117+ cells were identified the cells were prepared for differentiation and that was carried out using specific growth factors including BMP4, BMP7, FGF2, and retinoic acid, providing the niche to the stem cells for differentiation towards nephron progenitors which was confirmed by protein expression of Wilms Tumor-1 (WT1) using immunofluorescence analysis. The sample size for this invitro work was n=3. Results: We successfully isolated small percentage of CD117+ cells in amniotic fluid followed by in vitro expansion and differentiation towards nephron progenitor cells (NPCs) using well defined media and growth factors, initially differentiated cells were spindle shaped and showed fibroblastic appearance later at stage of nephron progenitors it attained the shape of rounded big clusters, differentiated cells stained positive for WT1 and negative for cluster of differentiation (CD117). Therefore, confirming the successful isolation and differentiation of amniotic fluid stem cells towards nephron progenitors. Conclusion: To the best of our knowledge this is the first study from the country on the use of Amniotic fluid stem cells and their differentiation towards nephron progenitors that can be used as substitution source of cell therapy for exploration of renal diseases at cellular and molecular level and potential regenerative medicine applications.

Application of a novel bacterial consortium BDAM for bioremediation of bispyribac sodium in wheat vegetated soil.

Plant-bacterial mutualism has tremendous potential for remediation of herbicide contaminated soils. Generally, bacterial inoculation helps plants to grow well in the contaminated environment. Here, we investigated the impact of bispyribac sodium (BS) degrading bacterial consortium (BDAM) on BS remediation, plant growth promotion and BS accumulation in plant parts. Wheat (Triticum aestivum) was planted in BS spiked soil and inoculated with BDAM. Inoculation showed a beneficial effect on plant biomass production and degradation of BS in the rhizosphere and the rhizosheath. After 40 and 60 days of inoculation, the degradation of BS was more than 96% and approximately 100% respectively in the planted and inoculated soil spiked with 2 and 5 mg kg-1 BS. However, in planted and un-inoculated soil, the degradation of BS was 72% after 60 days of sowing. Furthermore, inoculated bacterial strains colonized both in rhizo- and endosphere of the inoculated plants. In comparison with the un-inoculated soil, significantly less accumulation of BS was found in the roots and shoots of the plants growing in inoculated soil. We report the efficiency of plant-bacterial partnership for enhanced biodegradation of BS and to eliminate the BS residual toxicity to non-target plants.

Identification and analysis of 5-enolpyruvylshikimate-3-phosphate synthase (EPSPS) gene from glyphosate-resistant Ochrobactrum intermedium Sq20.

BACKGROUND: Glyphosate is a herbicide that acts by inhibition of the enzyme, 5-enolpyruvylshikimate-3-phosphate synthase (EPSPS), involved in the catalysis of an essential step in the biosynthesis of aromatic amino acids. The objective of this study was the isolation of glyphosate-resistant bacterial strains and subsequent characterization of the gene(s) encoding glyphosate resistance in these isolates. Using an enrichment culture technique, a glyphosate-resistant bacterium, Ochrobactrum intermedium Sq20 was isolated from glyphosate-contaminated indigenous soil and characterized. RESULTS: An open reading frame (ORF) comprising of 1353 bp potentially encoding aroAO. intermediumSq20 was amplified from O. intermedium Sq20. It showed 97% homology with aroA genes from other Ochrobactrum spp. Physicochemical characterization revealed that aroAO. intermediumSq20 encodes a polypeptide of 450 amino acids with a calculated molecular mass of 48.9782 kDa and an isoelectric point of 5.21. Secondary structure prediction of AroAO. intermediumSq20 demonstrated a high percentage of random coils and α helices. Methodical optimization and validation of the protein structure helped to build a reliable protein model indicating the presence of 91.8% amino acid residues in most favoured regions. In addition, strain Sq20 was found to be capable of complete degradation of glyphosate at 500 mg L-1 initial concentration as the sole carbon and energy source within 4 days. CONCLUSION: A glyphosate-resistant bacterial strain O. intermedium Sq20 was discovered. Sequence analysis and structure modelling demonstrated that AroAO. intermediumSq20 closely resembles class II EPSPS and possesses high glyphosate resistance. This provides a good foundation for functional analysis of experimentally derived crystal structures. The cloning and characterization of AroAO. intermediumSq20 will further help in understanding its role at the molecular level and its potential use in the production of glyphosate-resistant transgenic crops. © 2017 Society of Chemical Industry.

Enhanced remediation of chlorpyrifos by ryegrass (Lolium multiflorum) and a chlorpyrifos degrading bacterial endophyte Mezorhizobium sp. HN3.

For effective remediation of contaminants, plant-endophyte partnership is a promising field to be explored. Generally endophytic bacteria assist their host plant by withstanding the stress induced by the contaminants. The objective of this study was to explore the suitability of plant-bacterial partnership for chlorpyrifos (CP) remediation using ryegrass and a CP degrading endophyte, Mesorhizobium sp. HN3 which belongs to plant growth promoting rhizobia. The inoculated yfp-tagged Mesorhizobium sp. HN3 efficiently colonized in the rhizosphere, enhanced plant growth and degradation of CP and its metabolite 3,5,6 trichloro-2-pyridinol (TCP). Significantly lower CP residues were observed in the roots and shoots of plants vegetated in inoculated soil which might be attributed to the efficient root colonization of HN3yfp. These results suggest the involvement of Mesorhizobium sp. HN3yfp in CP degradation inside the roots and rhizosphere of plants and further emphasize on the effectiveness of endophytic bacteria in stimulating the remediation of pesticide contaminants. This is the first report which demonstrates the efficacy of bacterial endophyte for degradation of CP residues taken up by the plant and enhanced remediation of chlorpyrifos contaminated soil.