Clinical application of acellular matrix derived from the bubaline diaphragm and caprine rumen for the repair of abdominal wall defects in animals

The abdominal wall hernias resulting due to trauma or other clinical conditions are common in animals. Large hernias required the use of synthetic mesh, which is costly and may result in infection, fistula formation, and pain. Application of biomaterials in hernia repair causes a reduction in pain, reduced recovery time, and rate of recurrence. The study was undertaken to test the acellular bubaline diaphragm matrix (BDiaM) and acellular caprine rumen matrix (CRuM) for the repaired hernia in clinical cases. Fresh bubaline diaphragm and caprine rumen were decellularized using sodium deoxycholate (1% for CRuM and 2% for BDiaM) for 48h. Acellularity was ascertained histologically and by DNA quantification. Histologically, both the matrices showed complete acellularity and orderly arranged collagen fibers after 48 h. The DNA contents were significantly (P0.05) reduced in both the matrices in comparison to the native matrices. The BDiaM and CRuM matrices were applied in eight and nine clinical cases of abdominal wall defects, respectively. Animals with BDiaM and CRuM matrices recovered uneventfully and remained sound at least up to 3 months. Hematological and immunological findings were unremarkable. BDiaM and CRuM matrices showed good results without complications. Keywords:Biocompatibility, Bubaline diaphragm matrix, Caprine rumen matrix, DNA quantification, ELISA, SDS-PAG

Deforming Active Droplets in Viscoelastic Media

To mimic the motion of biological swimmers in bodily fluids, a novel experimental system of micellar solubilization driven active droplets in a visco-elastic polymeric solution is presented. The visco-elastic nature of the medium, characterized by the Deborah number (De), is tuned by varying the surfactant (fuel) and polymer concentration in the ambient medium. At moderate De, the droplet exhibits a steady deformed shape, markedly different from the spherical shape observed in Newtonian media. A theoretical analysis based on the normal stress balance at the interface is shown to accurately predict the droplet shape. With a further increase in De, time-periodic deformations accompanied by oscillatory transitions in swimming modes are observed. The study unveils the rich complexity in the motion of active droplets in viscoelastic fluids, which has been hitherto unexplored.Comment: 7 pages, 6 figures, 31 reference

Bubaline Cholecyst Derived Extracellular Matrix for Reconstruction of Full Thickness Skin Wounds in Rats

An acellular cholecyst derived extracellular matrix (b-CEM) of bubaline origin was prepared using anionic biological detergent. Healing potential of b-CEM was compared with commercially available collagen sheet (b-CS) and open wound (C) in full thickness skin wounds in rats. Thirty-six clinically healthy adult Sprague Dawley rats of either sex were randomly divided into three equal groups. Under general anesthesia, a full thickness skin wound (20 × 20 mm2) was created on the dorsum of each rat. The defect in group I was kept as open wound and was taken as control. In group II, the defect was repaired with commercially available collagen sheet (b-CS). In group III, the defect was repaired with cholecyst derived extracellular matrix of bovine origin (b-CEM). Planimetry, wound contracture, and immunological and histological observations were carried out to evaluate healing process. Significantly (P<0.05) increased wound contraction was observed in b-CEM (III) as compared to control (I) and b-CS (II) on day 21. Histologically, improved epithelization, neovascularization, fibroplasia, and best arranged collagen fibers were observed in b-CEM (III) as early as on postimplantation day 21. These findings indicate that b-CEM have potential for biomedical applications for full thickness skin wound repair in rats

Bone marrow derived cell-seeded extracellular matrix: A novel biomaterial in the field of wound management

Aim: Extensive or irreversible damage to the skin often requires additional skin substitutes for reconstruction. Biomaterials have become critical components in the development of effective new medical therapies for wound care. Materials and Methods: In the present study, a cell matrix construct (bone marrow-derived cells (BMdc) seeded extracellular matrix [ECM]) was used as a biological substitute for the repair of full-thickness skin wound. ECM was developed by decellularizing fish swim bladder (FSB). Goat bone marrow-derived cells (G-BMdc) were seeded over this decellularized matrix. Efficacy of this cell matrix construct in wound repair was tested by implanting it over 20 mm2 × 20 mm2 size fullthickness skin wound created over the dorsum of rat. The study was conducted in 16 clinically healthy adult rats of either sex. The animals were randomly divided into 2 equal groups of 8 animals each. In Group I, animal’s wounds were repaired with a cellular FSB matrix. In Group II, wounds were repaired with G-BMdc seeded a cellular FSB matrix. Immune response and efficacy of healing were analyzed. Results: Quality of healing and immuno tolerance to the biological substitute was significantly better in Group II than Group I. Conclusion: Seeding with BMdc increases the wound healing potency and modulates the immune response to a significantly negligible level. The BMdc seeded acellular FSB matrix was found to be a novel biomaterial for wound management

Activity of cefepime/zidebactam against MDR Escherichia coli isolates harbouring a novel mechanism of resistance based on four-amino-acid inserts in PBP3

BACKGROUND: Recent reports reveal the emergence of Escherichia coli isolates harbouring a novel resistance mechanism based on four-amino-acid inserts in PBP3. These organisms concomitantly expressed ESBLs or/and serine-/metallo-carbapenemases and were phenotypically detected by elevated aztreonam/avibactam MICs. OBJECTIVES: The in vitro activities of the investigational antibiotic cefepime/zidebactam and approved antibiotics (ceftazidime/avibactam, ceftolozane/tazobactam, imipenem/relebactam and others) were determined against E. coli isolates harbouring four-amino-acid inserts in PBP3. METHODS: Whole-genome sequenced E. coli isolates (n = 89) collected from a large tertiary care hospital in Southern India (n = 64) and from 12 tertiary care hospitals located across India (n = 25) during 2016-18, showing aztreonam/avibactam MICs ≥1 mg/L (≥4 times the aztreonam epidemiological cut-off) were included in this study. The MICs of antibiotics were determined using the reference broth microdilution method. RESULTS: Four-amino-acid inserts [YRIK (n = 30) and YRIN (n = 53)] were found in 83/89 isolates. Among 83 isolates, 65 carried carbapenemase genes [blaNDM (n = 39), blaOXA-48-like (n = 11) and blaNDM + blaOXA-48-like (n = 15)] and 18 isolates produced ESBLs/class C β-lactamases only. At least 16 unique STs were noted. Cefepime/zidebactam demonstrated potent activity, with all isolates inhibited at ≤1 mg/L. Comparator antibiotics including ceftazidime/avibactam and imipenem/relebactam showed limited activities. CONCLUSIONS: E. coli isolates concurrently harbouring four-amino-acid inserts in PBP3 and NDM are an emerging therapeutic challenge. Assisted by the PBP2-binding action of zidebactam, the cefepime/zidebactam combination overcomes both target modification (PBP3 insert)- and carbapenemase (NDM)-mediated resistance mechanisms in E. coli

Multiplexed Autoantibody Signature for Serological Detection of Canine Mammary Tumours

Abstract Spontaneously occurring canine mammary tumours (CMTs) are the most common neoplasms of female unspayed dogs and are of potential importance as models for human breast cancer as well. Mortality rates are thrice higher in dogs as compared to humans with breast cancer, which can partly be attributed to lack of diagnostic techniques for their early detection. Human breast cancer studies reveal role of autoantibodies in early cancer diagnosis and also the usefulness of autoantibody panels in increasing the sensitivity, as well as, specificity of diagnostic assays. Therefore, in this study, we took advantage of high-throughput Luminex technique for developing a multiplex assay to detect autoantibody signatures against 5 canine mammary tumour-associated autoantigens (TAAs). These TAAs were expressed separately as fusion proteins with halo tag at the N-terminus, which allows easy and specific covalent coupling with magnetic microspheres. The multiplex assay, comprising a panel of candidate autoantigens (TPI, PGAM1, MNSOD, CMYC & MUC1) was used for screening circulating autoantibodies in 125 dog sera samples, including 75 mammary tumour sera and 50 healthy dog sera. The area under curve (AUC) of the combined panel of biomarkers is 0.931 (p < 0.0001), which validates the discriminative potential of the panel in differentiating tumour patients from healthy controls. The assay could be conducted in 3hrs using only 1ul of serum sample and could detect clinical cases of canine mammary tumour with sensitivity and specificity of 78.6% and 90%, respectively. In this study, we report for the first time a multiplexed assay for detection of autoantibodies in canine tumours, utilizing luminex technology and halo-tag coupling strategy. Further to the best of our knowledge, autoantibodies to CMYC and MUC1 have been reported for the first time in canines in this study

Gene expression profiling of spontaneously occurring canine mammary tumours: Insight into gene networks and pathways linked to cancer pathogenesis.

Spontaneously occurring canine mammary tumours (CMTs) are the most common neoplasms of unspayed female dogs leading to thrice higher mortality rates than human breast cancer. These are also attractive models for human breast cancer studies owing to clinical and molecular similarities. Thus, they are important candidates for biomarker studies and understanding cancer pathobiology. The study was designed to explore underlying molecular networks and pathways in CMTs for deciphering new prognostic factors and therapeutic targets. To gain an insight into various pathways and networks associated with the development and pathogenesis of CMTs, comparative cDNA microarray expression profiling was performed using CMT tissues and healthy mammary gland tissues. Upon analysis, 1700 and 1287 differentially expressed genes (DEGs, P ≤ 0.05) were identified in malignant and benign tissues, respectively. DEGs identified from microarray analysis were further annotated using the Ingenuity Systems Pathway Analysis (IPA) tool for detection of deregulated canonical pathways, upstream regulators, and networks associated with malignant, as well as, benign disease. Top scoring key networks in benign and malignant mammary tumours were having central nodes of VEGF and BUB1B, respectively. Cyclins & cell cycle regulation and TREM1 signalling were amongst the top activated canonical pathways in CMTs. Other cancer related significant pathways like apoptosis signalling, dendritic cell maturation, DNA recombination and repair, Wnt/β-catenin signalling, etc. were also found to be altered. Furthermore, seven proteins (ANXA2, APOCII, CDK6, GATC, GDI2, GNAQ and MYH9) highly up-regulated in malignant tissues were identified by two-dimensional gel electrophoresis (2DE) and MALDI-TOF PMF studies which were in concordance with microarray data. Thus, the study has uncovered ample number of candidate genes associated with CMTs which need to be further validated as therapeutic targets and prognostic markers

Comparison of Tensile Bond Strength of Fixed-Fixed Versus Cantilever Single- and Double-Abutted Resin-Bonded Bridges Dental Prosthesis

Resin-bonded fixed dental prostheses (RBFDP) are minimally invasive alternatives to traditional full-coverage fixed partial dentures as they rely on resin cements for retention. This study compared and evaluated the tensile bond strength of three different resin-bonded bridge designs, namely, three-unit fixed-fixed, two-unit cantilever single abutment, and three-unit cantilever double-abutted resin-bonded bridge. Furthermore, the study attempted to compare the tensile bond strengths of the Maryland and Rochette types of resin-bonded bridges. Based on the inclusion and exclusion criteria, a total of seventy-five extracted maxillary incisors were collected and later were mounted on the acrylic blocks. Three distinct resin-bonded metal frameworks were designed: three-unit fixed-fixed (n = 30), two-unit cantilever single abutment (n = 30), and a three-unit cantilever double abutment (n = 30). The main groups were further divided into two subgroups based on the retainer design such as Rochette and Maryland. The different prosthesis designs were cemented to the prepared teeth. Later, abutment preparations were made on all specimens keeping the preparation as minimally invasive and esthetic oriented. Impression of the preparations were made using polyvinyl siloxane impression material, followed by pouring cast using die stone. A U-shaped handle of 1.5 mm diameter sprue wax with a 3 mm hole in between was attached to the occlusal surface of each pattern. The wax patterns were sprued and cast in a cobalt&ndash;chromium alloy. The castings were cleaned by sandblasting, followed by finishing and polishing. Lastly, based on the study group, specimens for Rochette bridge were perforated to provide mechanical retention between resin cement and metal, whereas the remaining 15 specimens were sandblasted on the palatal side to provide mechanical retention (Maryland bridge). In order to evaluate the tensile bond strength, the specimens were subjected to tensile forces on a universal testing machine with a uniform crosshead speed. The fixed-fixed partial prosthesis proved superior to both cantilever designs, whereas the single abutment cantilever design showed the lowest tensile bond strength. Maryland bridges uniformly showed higher bond strengths across all framework designs. Within the limitations of this study, the three-unit fixed-fixed design and Maryland bridges had greater bond strengths, implying that they may demonstrate lower clinical failure than cantilever designs and Rochette bridges