IDENTIFICATION OF CELLULOLYTIC BACTERIA ISOLATED FROM THE TERMITE COPTOTERMES CURVIGNATHUS (HOLMGREN)

The aim of the present research was to isolate and identify cellulolytic bacteria from the gut of the local termite Coptotermes curvignathus (Holmgren) present in the vicinity of the University of Putra Malaysia. The isolates were cultured in a medium containing carboxymethyl-cellulose and cellobiose. The bacterial species were tentatively identified by using the Biolog reader as well as the Bergey's manual and later confirmed by 16S rRNA sequence homology. The species were all novel strains and identified as Bacillus cereus strain Razmin A, Enterobacter aerogenes strain Razmin B, Enterobacter cloacae strain Razmin C, Chryseobacterium kwangyangense strain Cb and Acinetobacter strain Raminalimon. Biolog reader was not able to identify one of the bacterial species in which it was identified as C. kwangyangense strain Cb by 16S rRNA sequence homology. The GenBank NCBI/EMBL accession numbers for the bacterial strains are EU294508, EU305608, EU305609, EU169201 and EU332791 for B. cereus Razmin A, E. aerogenes strain Razmin B, E. cloacae strain Razmin C, C. kwangyangense strain Cb and Acinetobacter strain Raminalimon, respectively.

PRACTICAL APPLICATIONS

The experiment describes the isolation and identification of cellulolytic bacteria from termites, using Biolog reader and polymerase chain reaction technique. Termites are common insects in tropical regions and thrive on wood and cellulolytic materials. Hence, the bacteria isolated may be useful in the degradation of cellulosic materials to increase their digestibility and possibly production of metabolites and enzymes.     

Mineral phase in linguloid brachiopod shell: Lingula adamsi

The linguloid brachiopod shell family has been the focus of several studies because of the similarity in the composition of the mineral phase of these shells to that of human bone. However, ultrastructural features of Lingula shells have not yet been fully demonstrated at high magnification using Transmission Electron Microscopy (TEM) and Electron Diffraction. Ultrastructural characterization of the mineral phase in Lingula shells will improve our understanding of the biomineralization processes and mineral/organic interaction in more complex systems such as in bone or in other human mineralized tissues. In this study, the mineral phase of Lingula adamsi was characterized using a combination of ultrastructural and crystallographic techniques. The results showed that L. adamsi shells consist of apatite crystals of varying size, shape, and orientation in different areas of the shell. The c-axis of apatite was parallel to the shell surface and crystals were organized in different laminae. Compared to trabecular bovine bone, L. adamsi shells demonstrated a higher crystallinity and a lower amount of carbonate and organic compounds. This study therefore demonstrated how dissimilar organic matrix between L. adamsi shell and trabecular bone can modify the ultrastructural characteristics of apatite crystals in these two biomineralized tissues.     

Translation of a standardized manufacturing protocol for mesenchymal stromal cells: A systematic comparison of validation and manufacturing data

BackgroundMany data are available on expansion protocols for mesenchymal stromal cells (MSCs) for both experimental settings and manufacturing for clinical trials. However, there is a lack of information on translation of established protocols for Good Manufacturing Practice (GMP) from validation to manufacturing for clinical application. We present the validation and translation of a standardized pre-clinical protocol for isolation and expansion of MSCs for a clinical trial for reconstitution of alveolar bone.MethodsKey parameters of 22 large-scale expansions of MSCs from bone marrow (BM) for validation were compared with 11 expansions manufactured for the clinical trial “Jaw bone reconstruction using a combination of autologous mesenchymal stromal cells and biomaterial prior to dental implant placement (MAXILLO1)” aimed at reconstruction of alveolar bone.ResultsDespite variations of the starting material, the robust protocol led to stable performance characteristics of expanded MSCs. Manufacturing of the autologous advanced therapy medicinal product MAXILLO-1-MSC was possible, requiring 21 days for each product. Transport of BM aspirates and MSCs within 24 h was guaranteed. MSCs fulfilled quality criteria requested by the national competent authority. In one case, the delivered MSCs developed a mosaic in chromosomal finding, showing no abnormality in differentiation capacity, growth behavior or surface marker expression during long-term culture. The proportion of cells with the mosaic decreased in long-term culture and cells stopped growth after 38.4 population doublings.ConclusionsClinical use of freshly prepared MSCs, manufactured according to a standardized and validated protocol, is feasible for bone regeneration, even if there was a long local distance between manufacturing center and clinical site. Several parameters, such as colony forming units fibroblasts (CFU-F), percentage of CD34+ cells, cell count of mononuclear cells (MNCs) and white blood cells (WBCs), of the BM may serve as a predictive tool for the yield of MSCs and may help to avoid unnecessary costs for MSC manufacturing due to insufficient cell expansion rates.