Delineating the mechanistic relevance of the TP53 gene and its mutational impact on gene expression and patients' survival in bladder cancer.

Bladder carcinoma (BLCA) is a widespread urological malignancy causing significant global mortality, often hindered by delayed diagnosis and limited treatments. BLCA frequently exhibits TP53 mutations, playing a pivotal role in its pathogenesis and underscoring the potential of targeting TP53 as a therapeutic approach for this prevalent urological malignancy. Tumor tissues from 50 bladder cancer patients were used for mutational analysis in TP53's mutation-rich exons (5, 7, & 8). The gene expression of the TP53 gene, along with a TP53-target gene B-cell translocation gene 2 (BTG2) was also assessed in the cDNA samples from the same BLCA tissues and 15 urine controls of healthy people. The analysis revealed 22 % of patients with somatic hotspot mutations, 18 % with pathogenic missense mutations, and 12 % with intronic variants. Patients with somatic mutations exhibited the worst prognosis, supported by survival analysis from The Cancer Genome Atlas (TCGA) BLCA data. Interestingly, H296Y missense mutation correlated with higher TP53 expression and improved survival, while intronic SNPs were linked to worse outcomes. Additionally, upregulated BTG2 expression in mutated patients was observed which was correlated with poor prognosis, emphasizing the role of TP53 mutations in bladder cancer progression. The multivariate analysis highlighted the predictive power of TP53 mutations, with a high frequency of high-grade tumors (78.57 %) in mutated patients, underscoring their role in cancer progression. In conclusion, this study emphasizes the crucial role of TP53 mutations in bladder cancer patients from Bangladesh.

Computational exploration of SLC14A1 genetic variants through structure modeling, protein-ligand docking, and molecular dynamics simulation.

The urea transporter UT-B1, encoded by the SLC14A1 gene, has been hypothesized to be a significant protein whose deficiency and dysfunction contribute to the pathogenesis of bladder cancer and many other diseases. Several studies reported the association of genetic alterations in the SLC14A1 (UT-B1) gene with bladder carcinogenesis, suggesting a need for thorough characterization of the UT-B1 protein's coding and non-coding variants. This study used various computational techniques to investigate the commonly occurring germ-line missense and non-coding SNPs (ncSNPs) of the SLC14A1 gene (UT-B1) for their structural, functional, and molecular implications for disease susceptibility and dysfunctionality. SLC14A1 missense variants, primarily identified from the ENSEMBL genome browser, were screened through twelve functionality prediction tools leading to two variants D280Y (predicted detrimental by maximum tools) and D280N (high global MAF) for rs1058396. Subsequently, the ConSurf and NetSurf tools revealed the D280 residue to be in a variable site and exposed on the protein surface. According to I-Mutant2.0 and MUpro, both variants are predicted to cause a significant effect on protein stability. Analysis of molecular docking anticipated these two variants to decrease the binding affinity of UT-B1 protein for the examined ligands to a significant extent. Molecular dynamics also disclosed the possible destabilization of the UT-B1 protein due to single nucleotide polymorphism compared to wild-type protein which may result in impaired protein function. Furthermore, several non-coding SNPs were estimated to affect transcription factor binding and regulation of SLC14A1 gene expression. Additionally, two ncSNPs were found to affect miRNA-based post-transcriptional regulation by creating new seed regions for miRNA binding. This comprehensive in-silico study of SLC14A1 gene variants may serve as a springboard for future large-scale investigations examining SLC14A1 polymorphisms.

Identification and validation of prognostic signature genes of bladder cancer by integrating methylation and transcriptomic analysis.

Being a frequent malignant tumor of the genitourinary system, Bladder Urothelial Carcinoma (BLCA) has a poor prognosis. This study focused on identifying and validating prognostic biomarkers utilizing methylation, transcriptomics, and clinical data from The Cancer Genome Atlas Bladder Urothelial Carcinoma (TCGA BLCA) cohort. The impact of altered differentially methylated hallmark pathway genes was subjected to clustering analysis to observe changes in the transcriptional landscape on BLCA patients and identify two subtypes of patients from the TCGA BLCA population where Subtype 2 was associated with the worst prognosis with a p-value of 0.00032. Differential expression and enrichment analysis showed that subtype 2 was enriched in immune-responsive and cancer-progressive pathways, whereas subtype 1 was enriched in biosynthetic pathways. Following, regression and network analyses revealed Epidermal Growth Factor Receptor (EGFR), Fos-related antigen 1 (FOSL1), Nuclear Factor Erythroid 2 (NFE2), ADP-ribosylation factor-like protein 4D (ARL4D), SH3 domain containing ring finger 2 (SH3RF2), and Cadherin 3 (CDH3) genes to be the most significant prognostic gene markers. These genes were used to construct a risk model that separated the BLCA patients into high and low-risk groups. The risk model was also validated in an external dataset by performing survival analysis between high and low-risk groups with a p-value < 0.001 and the result showed the high group was significantly associated with poor prognosis compared to the low group. Single-cell analyses revealed the elevated level of these genes in the tumor microenvironment and associated with immune response. High-grade patients also tend to have a high expression of these genes compared to low-grade patients. In conclusion, this research developed a six-gene signature that is pertinent to the prediction of overall survival (OS) and might contribute to the advancement of precision medicine in the management of bladder cancer.

Comprehensive characterization of coding and non-coding single nucleotide polymorphisms of the Myoneurin (MYNN) gene using molecular dynamics simulation and docking approaches.

Genome-wide association studies (GWAS) identified a coding single nucleotide polymorphism, MYNN rs10936599, at chromosome 3q. MYNN gene encodes myoneurin protein, which has been associated with several cancer pathogenesis and disease development processes. However, there needed to be a more detailed characterization of this polymorphism's (and other coding and non-coding polymorphisms) structural, functional, and molecular impact. The current study addressed this gap and analyzed different properties of rs10936599 and non-coding SNPs of MYNN via a thorough computational method. The variant, rs10936599, was predicted functionally deleterious by nine functionality prediction approaches, like SIFT, PolyPhen-2, and REVEL, etc. Following that, structural modifications were estimated through the HOPE server and Mutation3D. Moreover, the mutation was found in a conserved and active residue, according to ConSurf and CPORT. Further, the secondary structures were predicted, followed by tertiary structures, and there was a significant deviation between the native and variant models. Similarly, molecular simulation also showed considerable differences in the dynamic pattern of the wildtype and mutant structures. Molecular docking revealed that the variant binds with better docking scores with ligand NOTCH2. In addition to that, non-coding SNPs located at the MYNN locus were retrieved from the ENSEMBL database. These were found to disrupt the transcription factor binding regulatory regions; nonetheless, only two affect miRNA target sites. Again, eight non-coding variants were detected in the testes with normalized expression, whereas HaploReg v4.1 unveiled annotations for non-coding variants. In summary, in silico comprehensive characterization of coding and non-coding single nucleotide polymorphisms of MYNN gene will assist researchers to work on MYNN gene and establish their association with certain types of cancers.

Physicochemical, in vitro and in vivo evaluation of VEGF loaded PCL-mPEG and PDGF loaded PCL-Chitosan dual layered vascular grafts.

The present study has attempted to evaluate the endothelialization and smooth muscle regeneration efficiency of a novel dual-layer small-diameter vascular graft. Two types of layers (PCL-mPEG-VEGF and PCL-Chitosan-PDGF) were fabricated to find out the best layer giving endothelialization support for the lumen and unique contractile function for outer layer of blood vessels. Platelet-derived growth factor (PDGF) and chitosan were immobilized onto PCL surface by aminolysis-based surface modification technique. Besides, Poly (ethylene glycol) methyl ether (mPEG) and vascular endothelial growth factor (VEGF) were directly blended with PCL. Morphological analysis of membranes ensured consistency of average fibers diameter with native extracellular matrix. A favorable interaction of PCL-mPEG-VEGF with cow pulmonary endothelial cells (CPAEs) and PCL-Chitosan-PDGF with rat bone marrow mesenchymal stem cells (RBMSCs) was obtained during in vitro study. Controlled growth factor release patterns were found from both layers. Further, PCL-mPEG-VEGF exhibited endothelial markers expression properties from RBMSCs. Up-regulation of SMCs markers expression was significantly ensured by the PCL-Chitosan-PDGF membrane. Thus, PCL-mPEG-VEGF and PCL-Chitosan-PDGF were preferred as inner and outer layers respectively of a finally prepared tubular hybrid tissue engineered small diameter vascular graft. Finally, the dual-layer vascular graft was implanted onto a rat abdominal aorta model for 2 months. The extracted samples exhibited the presence of endothelial marker (ICAM 1) in the inner layer and smooth muscle cell marker (αSMA) in the outer layer as well as substantial amount of collagen deposition was observed in the both layers.

Liver tissue-derived ECM loaded nanocellulose-alginate-TCP composite beads for accelerated bone regeneration.

Guided bone regeneration with osteoinductive scaffolds is a competitive edge of tissue engineering due to faster and more consistent healing. In the present study, we developed such composite beads with nanocellulose reinforced alginate hydrogel that carriedß-tricalcium phosphate (ß-TCP) nano-powder and liver-derived extracellular matrix (ECM) from porcine. Interestingly, it was observed that the beads' group containing ECM-ß-TCP-alginate-nanocellulose (ETAC) was more cytocompatible than the others comprised ofß-TCP-alginate-nanocellulose (TAC) and alginate-nanocellulose (AC). Cell attachment on ETAC beads was dramatically increased with time. In parallel within vitroresults, ETAC beads produced uniform cortical and cancellous bone in the femur defect model of rabbits within 2 months. Although the group TAC also produced noticeable bone in the defect site, the healing quality was improved and regeneration was faster after adding ECM. This conclusion was not only confirmed by micro-anatomical analysis but also demonstrated with x-ray microtomography. In addition, the characteristic moldable and injectable properties made ETAC a promising scaffold for clinical applications.

Calibration and field validation of POCIS passive samplers for tracking artificial sweeteners as indicators of municipal wastewater contamination in surface waters.

Polar organic chemical integrative samplers (POCIS) are widely used to track contaminants in surface waters. However, POCIS have not been used previously to monitor for artificial sweeteners as an indicator of wastewater pollution. In this study, we report for the first time the POCIS sampling rates (Rscal) for four artificial sweetener compounds, acesulfame (0.001 L/day), sucralose (0.114 L/day), cyclamate (0.001 L/day), and saccharin (0.002 L/day). We also prepared a modified POCIS with Strata X-AW anion exchange resin as a sorbent (i.e., ax-POCIS) and determined the sampling rates for sucralose (0.060 L/day) and acesulfame (0.128 L/day). Rscal values were adjusted according to the rate of loss of the performance reference compound, metoprolol-d6 from deployed POCIS to yield field sampling rates (i.e., Rsfield). Field validation of the monitoring method was conducted in Presqu'ile Bay on the north-central coast of Lake Ontario that is impacted by discharges from a sewage lagoon. POCIS were deployed at four sites within the bay and in the lagoon discharge. The four artificial sweeteners, as well as caffeine, ibuprofen, and other microcontaminants of sewage origin, were present throughout the bay at estimated concentrations in the ng/L range, and in the lagoon discharge at estimated concentrations higher by approximately one order of magnitude. Because acesulfame is present in ionic form over the pH range of natural waters, there are uncertainties related to the sampling rates using the standard POCIS. Sucralose is recommended as the best choice for source tracking using POCIS. There was good agreement between the concentrations of sucralose estimated from POCIS and the measured concentrations in grab samples of surface water in the bay. The present study provides key data for monitoring artificial sweeteners using POCIS.

Sources of microbial contamination in the watershed and coastal zone of Soufriere, St. Lucia.

The Sustainable Development Goal 6 calls for global progress by 2030 in treating domestic wastewater and providing access to adequate sanitation facilities. However, meeting these goals will be a challenge for most Small Island Developing States, including Caribbean island nations. In the nearshore zone of the Soufriere region on the Caribbean island of St. Lucia, there is a history of high levels of bacteria of fecal origin. Possible land-based sources of microbial contamination in the Soufriere Bay include discharges from the Soufriere River and transport of wastewater, including fecal material from the town of Soufriere. This area is an important tourist destination and supports a local fishery. To identify the sources of microbial contamination in Soufriere Bay, a range of monitoring methods were employed in this study. In grab samples of surface water collected from the Soufriere River, counts of total coliforms and Escherichia coli were elevated above water quality guidelines. However, the spikes in concentrations of these indicator organisms in the river did not necessarily coincide with the spikes in the levels of total coliforms and E. coli detected in samples collected on the same dates in Soufriere Bay, indicating that there are other sources of pollution in the Bay besides discharges from the river. Monitoring for chemical indicators of wastewater (i.e., caffeine, sucralose, fluconazole) in the Soufriere River indicated that there are inputs of sewage or human fecal material throughout the watershed. However, analysis of Bacteroidales 16S rRNA genetic markers for fecal bacteria originating from humans, bovine ruminants, or other warm-blooded animals indicated that the majority of microbial contamination in the river was not from humans. Monitoring for chemical indicators of wastewater using passive samplers deployed in Soufriere Bay indicated that there are two "hot spots" of contamination located offshore of economically depressed areas of the town of Soufriere. This study indicates that efforts to control contamination of Soufriere Bay by fecal microorganisms must include management of pollution originating from both sewage and domestic animals in the watershed.

Clothianidin interferes with recognition of a previous encounter in rusty crayfish (Faxonius rusticus) due to a chemosensory impairment.

Clothianidin, a neonicotinoid insecticide that binds to arthropod nicotinic acetylcholine receptors, is widely used to protect plants against a wide variety of agricultural pests. Little is known about how this insecticide affects non-target invertebrate species in aquatic environments. In this study, we explored the effects of aqueous exposures of clothianidin on locomotion, chemosensory-based responses, and agonistic encounters of rusty crayfish (Faxonius rusticus). Clothianidin exposures at a concentration of 1.0 µg/L (i.e., 1.0 ppb) did not alter initiations and retreats, but did increase the amount of time the crayfish interacted per interaction. In a subsequent food cue experiment with crayfish exposed to clothianidin concentrations of 0.4 µg/L and 1.0 µg/L, the test organisms demonstrated chemosensory dysfunction, but no decrease in locomotory movement. As chemosensation is essential for recognizing previous rivals in crayfish, the loss of this sense likely resulted in the exposed crayfish being unable to detect cues used to recognize a previous competitor. An inability to recognize a previous competitor (and who won or lost the previous interaction) could result in crayfish spending more time fighting and less time on foraging and reproduction. This study demonstrates that exposures of crayfish to clothianidin at concentrations found in the environment affects the behavioural ecology of these aquatic invertebrates.

Fabrication of thrombin loaded TEMPO-oxidized cellulose nanofiber-gelatin sponges and their hemostatic behavior in rat liver hemorrhage model.

Excessive blood loss due to trauma or major surgical intervention can be life threatening which necessitates rapid hemorrhage management for the prevention of such bleeding related sufferings. Broad interest in developing new hemostatic technologies have been paid for bleeding control but none of them found completely satisfactory especially in terms of rapid clotting, absorbability, porosity, cost effectiveness and safety. To address these issues, a combination of active and passive hemostatic materials from biological sources could be a wise choice. Therefore, plant-derived TEMPO-oxidized nanocellulose (TOCN)/biopolymer gelatin (G) sponge was successfully prepared in co-operation with intrinsic blood coagulation enzyme thrombin (Th) via freeze drying method and their application as rapid hemostatic dressing was investigated. Morphological and in vitro characteristics of the samples were evaluated where uniformity, porosity, swelling, degradation behavior had direct relationship with the percent gelatin incorporation. In vitro hemocompatibility and cyto-compatibility of these sponges were confirmed as well. Among the samples, TOCN 2.5G-Th sponge exhibited excellent hemostatic effect, rapid absorbability, minimum clotting time (1.37 ± 0.152 min) and reduction of blood loss was ensured through rat liver punch biopsy model. The results demonstrated that, Th enhanced blood coagulation, platelet and red blood cell aggregation following application of biopolymer TOCN 2.5G-Th sponge compared with samples devoid of Th. In short, the functional, cost effective and nontoxic sponge developed via facile preparation could potentially be used as an absorbable biomaterial to achieve immediate hemostasis. HighlightsPlant-derived TEMPO-oxidized nanocellulose (TOCN) and biopolymer gelatin (G) was successfully used to prepare a hemostatic sponge in combination with intrinsic blood coagulation enzyme thrombin (Th).The TG sponge combines the advantages of TOCN and gelatin, exhibiting biocompatibility, biodegradability and superior blood-absorption performance.The TOCN 2.5G-Th sponge improves plasma absorption, red blood cell adhesion, aggregation, platelet adhesion and activation leading to enhanced hemostasis effect and shorter hemostasis time in vitro and in vivo.

Multi-functional nanocellulose-chitosan dressing loaded with antibacterial lawsone for rapid hemostasis and cutaneous wound healing.

Cutaneous wounds accompanied by massive bleeding, bacterial infections might be lethal and cause fundamental therapeutic impediments in clinical fields. As part of the push for a solution, biomaterial having hemostatic-antibacterial features is highly desirable. Inspired by this concept, freeze dried sponges were developed followed by combining tempo-oxidized nanocellulose (TOCN), chitosan using EDC/NHS cross-linker with antibacterial lawsone loading for controlled delivery of this compound during wound healing. The pore diameter decreased upon increasing chitosan (2.5, 3.5, 4.5, 5.5% w/v) while TOCN ensured scaffold's mechanical stability. The in vitro degradation, lawsone release from fibroblast cell-compatible sponge was faster in acidic pH 5.5 than physiologic pH 7.4 indicating adaptability to physiological skin milieu of wounds. The rat tail amputation model, 14 days rat full-thickness cutaneous-wound model ensured hemostasis, dramatic wound closure after TLC4.5 (optimized scaffold) treatment suggesting its potential as functional wound healing substitute showing obvious avenue for hemostatis and skin tissue reconstruction arena.

Nocardia infection causing non-healing surgical wounds: A case series from Bangladesh.

OBJECTIVE: Nocardia can be introduced accidentally causing non-healing surgical wounds. METHOD: From February 2017 to January 2021 samples from wounds were collected. Nocardia identification and susceptibility testing were carried out by standard procedure. RESULTS: Seventeen (35.4 %) Nocardia spp. and 20 other pathogens (41.7%) were recovered by culture. Drug susceptibility among Nocardia was >70% to amikacin, amoxicillin-clavulanic acid, levofloxacin, linezolid, and imipenem, 47% to cephalosporins and 41% to trimethoprim/sulfamethoxazole. CONCLUSIONS: Infections with Nocardia spp. should be considered in non-healing surgical wounds.

Hematological abnormalities and comorbidities are associated with COVID-19 severity among hospitalized patients: Experience from Bangladesh.

BACKGROUND: The hematological abnormalities are assumed to be involved in the disease progression of COVID-19. However, the actual associations between specific blood parameters and COVID-19 are not well understood. Here we aimed to assess the correlations between hematological parameters and the severity of COVID-19. METHODS: We included COVID-19 patients who were admitted to Evercare Hospital Ltd, Dhaka, Bangladesh, between November 10, 2020, to April 12, 2021, with a confirmed case of RT-PCR test. We recorded demographic information, clinical data, and routine hematological examination results of all COVID-19 patients. We performed statistical analyses and interpretation of data to compare severe COVID-19 patients (SCP) and non-severe COVID-19 patients (NSCP). RESULTS: The age and BMI of the admitted COVID-19 patients were 48.79±8.53 years and 25.82±3.75 kg/m2. This study included a total of 306 hospitalized COVID-19 patients. Among them, NSCP and SCP were 198 and 108, respectively. And we recorded 12 deaths from SCP. We observed the alterations of several hematological parameters between SCP and NSCP. Among them, we noticed the increased levels of C-reactive protein (CRP), d-dimer, and ferritin showed good indicative value to evaluate the severity of COVID-19. Also, there were positive correlations among these parameters. Moreover, we found correlations between the outcomes of COVID-19 patients with patient's demographics and comorbid diseases. CONCLUSION: Based on our results, CRP, d-dimer, and ferritin levels at admission to hospitals represent simple assessment factors for COVID-19 severity and the treatment decisions at the hospital setup. These blood parameters could serve as indicators for the prognosis and severity of COVID-19. Therefore, our study findings might help to develop a treatment protocol for COVID-19 patients at the hospital setup.

Effects of opioids on reproduction in Japanese medaka, Oryzias latipes.

To study the effects of exposure of fish to opioid drugs, we exposed Japanese medaka (Oryzias latipes) over a full life cycle to codeine spiked into river water at nominal concentrations of 100, 1,000 and 25,000 ng/L and to fentanyl spiked into river water at nominal concentrations of 5, 25 and 1,000 ng/L. The measured concentrations during medaka exposures were consistent with the nominal concentrations. Treatments with codeine at all test concentrations reduced the number of eggs produced by female medaka, as well as the number of mature oocytes observed histologically in the ovaries. Exposures to codeine also resulted in altered concentrations of hormones within the hypothalamic-pituitary-gonadal axis, including reduced levels of 17ß-estradiol in female medaka. Fentanyl did not affect reproduction or the levels of hormones in medaka at the concentrations tested. Monitoring of surface waters in southern Ontario, Canada downstream of wastewater treatment plants showed that the test concentrations of fentanyl and codeine were environmentally relevant. The results of this work contribute to the literature on the impacts of opioids and other drugs of abuse released into surface waters.

Polycaprolactone-gelatin membrane as a sealant biomaterial efficiently prevents postoperative anastomotic leakage with promoting tissue repair.

Anastomotic leakage due to post-surgical suture line disruption is one of the crucial factors affecting patient's survival and quality of life. To resolve the poor healing of surgical anastomosis and protect suture sites leakage, fibrous membrane sealing patch was developed using a synthetic polymer (polycaprolactone (PCL)) and biopolymer (gelatin). Electrospinning was used to develop fibrous architecture of membranes fabricated in different ratios (15% (w/v) PCL: 15% (w/v) gelatin mixing ratio of 1:1, 1:2, 1:3 and 1:4). Experimental findings suggested that, higher gelatin content in the membranes reduced the fiber diameter and contact angle, leading to a more hydrophilic scaffold facilitating attachment to the defect site. The degradation rate of various PCL-gelatin membranes (P1G1, P1G2, P1G3 and P1G4) was proportional to the gelatin content. Cytocompatibility was assessed using L929 cells while the P1G4 (PCL: gelatin 1:4 ratio) scaffold exhibited optimum outcome. From in vivo study, the wound site healed significantly without any leakage when the sutured area of rat caecum was covered with P1G4 membrane whereas rats in the control group (suture only) showed leakage after two weeks of surgery. In summary, the P1G4 membrane has potential to be applied as a post-surgical leakage-preventing tissue repair biomaterial.

Fibroblast cell derived extracellular matrix containing electrospun scaffold as a hybrid biomaterial to promote in vitro endothelial cell expansion and functionalization.

Biomaterials to be used for vascular tissue engineering must allow attachment, proliferation, and functionalization of vasoactive cells especially endothelial cells. In this study, decellularized L929 fibroblast cell-derived ECM containing electrospun scaffolds were fabricated and their biological response was investigated using rat glomerulus endothelial cells (rGECs). The L929 cells were grown for one week to get cell sheets on PCL membranes followed by decellularization of whole cell sheet-PCL membrane (PCL-ECM) using sodium dodecyl sulfate (SDS)/triton X-100 (TX) or freeze/thaw (F/T)/Deoxyribonuclease cycle to yield the corresponding mechanically stable scaffold. The nucleic acids and structural proteins quantification were performed on various membranes before and after decellularization process. Seeded rGECs on PCL, PCL-ECM (SDS/TX) and PCL-ECM (F/T) membranes were investigated through immunofluorescence and cell proliferation assay. The bio-macromolecules contents on decellularized scaffolds showed diverse outcome because of different decellularization methods used. The hydrophilic PCL-ECM (F/T) scaffold showed the best result by ensuring stability, good cytocompatibility, and interconnections among endothelial cells as was further confirmed by endothelial gene expression analysis. In short, the outcomes of this study may pave the way for the construction of new cell-derived ECM based vascular tissue engineering scaffolds as well as for the development of in vitro models to study endothelial cell function.

Boosting osteogenic potential and bone regeneration by co-cultured cell derived extracellular matrix incorporated porous electrospun scaffold.

Implants for bone regeneration to remedy segmental bone defects, osteomyelitis, necrotic bone tissue and non-union fractures have worldwide appeal. Although biomaterials offer most of the advantages by improving tissue growth but developments are more commonly achieved via biologically derived molecules. To aid site specific bone tissue regeneration by synthetic scaffold, cell derived extracellular matrix (ECM) can be a crucial component. In this study, co-cultured bone marrow mesenchymal stem cell and osteoblastic cells derived ECM incorporated electrospun polycaprolactone (PCL) membranes were assessed for bone tissue engineering application. The preliminary experimental details indicated that, co-culture of cells supported enhanced in vitro ECM synthesis followed by successful deposition of osteoblastic ECM into electrospun membranes. The acellular samples revealed retention of ECM related biomacromolecules (collagen, glycosaminoglycan) and partial recovery of pores after decellularization. In vitro biocompatibility tests ensured improvement of proliferation and osteoblastic differentiation of MC3T3-E1 cells in decellularized ECM containing membrane (PCL-ECM) compared to bare membrane (PCL-B) which was further confirmed by osteogenic marker proteins expression analysis. The decellularized PCL-ECM membrane allowed great improvement of bone regeneration over the bare membrane (PCL-B) in 8 mm size critical sized rat skull defects at 2 months of post implantation. In short, the outcome of this study could be impactful in development and application of cell derived ECM based synthetic electrospun templates for bone tissue engineering application.[Formula: see text].

Dissipation of sulfamethoxazole and trimethoprim during temporary storage of biosolids: A microcosm study.

Little is known about the dissipation rate of microcontaminants in biosolids during storage and stabilization in stockpiles (unsaturated) or storage lagoons/tanks (saturated). The objective of this study was to characterize the dissipation in biosolids of two antibiotics, sulfamethoxazole (SMX) and trimethoprim (TMP), in microcosms under saturated and unsaturated conditions that simulate biosolids that are stockpiled on land or deposited in lagoons/tanks, respectively. The laboratory experiment was conducted at 22 °C using biosolids spiked at an initial nominal concentration of 10 mg kg-1 for both antibiotics. Biosolids were sampled in triplicate at seven sampling times over a 42-d period. Concentrations of SMX and TMP in extracts prepared from biosolids were quantified using liquid chromatography with tandem mass spectrometry. Dissipation data fitted to a first-order kinetic model indicated that the time to 50% dissipation (DT50) for SMX was significantly shorter in the unsaturated microcosms (2.8 d) than the saturated microcosms (4.4 d), while the DT50 for TMP was significantly shorter in microcosms under saturated conditions (10 d) relative to unsaturated conditions (116 d). These results indicate that the reducing conditions that develop in biosolids deposited in lagoons or placed in storage tanks might be effective for enhancing the microbial degradation of antibiotics that are otherwise persistent under aerobic conditions (i.e., TMP), while also being effective for removing other antibiotics including those that dissipate relatively readily under aerobic conditions (i.e., SMX).

Thermal stimuli-responsive hyaluronic acid loaded cellulose based physical hydrogel for post-surgical de novo peritoneal adhesion prevention.

Effective strategies for post-surgical adhesion prevention have increasingly focused on injectable adhesion barriers due to their minimal invasiveness and wider applicability. In this study, a thermo-reversible hydrogel was developed by combining high molecular weight hyaluronic acid (HA) at various concentrations (0.05, 0.25, and 0.45% w/v) with tempo-oxidized nanocellulose (TOCN), methyl cellulose (MC) and polyethylene glycol (PEG) for anti-adhesion application. The hydrogel preparation time was short and did not require any chemical modification. TOCN ensured the mechanical stability of the hydrogel. MC confirmed thermo-sensitive feature. Higher amounts of HA increased the rate of hydrogel degradation. The HA 0.25 hydrogel was free-flowing, injectable at ambient temperature, capable of faster (40 ± 2 s), and reversible sol-gel (4 °C-37 °C) transition. A rat side-wall cecum abrasion model was used to confirm the complete de novo adhesion prevention efficacy of optimized HA 0.25 hydrogel, where the scratched abdominal wall of animals treated with HA 0.25 hydrogel healed after 14 days. During in vivo experiment, PEG in the hydrogel played a crucial role in adhesion prevention by minimizing friction between the surgical site and nearby organs. In a nutshell, HA 0.25 hydrogel, fabricated without crosslinking agent, is a potential candidate for tissue adhesion prevention strategies.