Low background mould-prepared gelatine standards for reproducible quantification in elemental bio-imaging.

Standard preparation for elemental bio-imaging by laser ablation-inductively coupled plasma-mass spectrometry is confounded by the chemical and physical differences between standard and sample matrices. These differences lead to variable ablation, aerosol generation and transportation characteristics and must be considered when designing matrix-matched standards for reliable calibration and quantification. The ability to precisely mimic sample matrices is hampered due to the complexity and heterogeneity of biological tissue and small variabilities in standard matrices and sample composition often negatively impact accuracy, precision and robustness. Furthermore, cumbersome preparation protocols may limit reproducibility and traceability. This work presents novel facile methods for the preparation of gelatine standards using both commercial and laboratory-made moulds. Surface roughness, thickness and robustness of the mould-prepared standards were compared against cryo-sectioned gelatine and homogenised brain tissue standards. The mould-prepared standards had excellent thickness accuracy and signal precision which allowed robust quantification, were easier to prepare and therefore easier to reproduce. We also compared gelatine standards prepared from a variety of animal sources and discuss their suitability to calibrate low level elemental concentrations. Finally, we present a simple method to remove background metals in gelatine using various chelating resins to increase the dynamic calibration range and to improve limits of analysis.

Quantitative Determination and Subcellular Imaging of Cu in Single Cells via Laser Ablation-ICP-Mass Spectrometry Using High-Density Microarray Gelatin Standards.

This manuscript describes the development and characterization of a high-density microarray calibration standard, manufactured in-house and designed to overcome the limitations in precision, accuracy, and throughput of current calibration approaches for the quantification of elemental concentrations on the cellular level using laser ablation-inductively coupled plasma-mass spectrometry (LA-ICPMS). As a case study, the accumulation of Cu in the model organism Scrippsiella trochoidea resulting from transition metal exposure (ranging from 0.5 to 100 µg/L) was evaluated. After the Cu exposure, cells of this photosynthetic dinoflagellate were treated with a critical point drying protocol, transferred to a carbon stub, and sputter-coated with a Au layer for scanning electron microscopy (SEM) analysis. In subsequent LA-ICPMS analysis, approximately 100 cells of each population were individually ablated. This approach permitted the evaluation of the mean concentration of Cu in the cell population across different exposure levels and also allowed the examination of the cellular distribution of Cu within the populations. In a cross-validation exercise, subcellular LA-ICPMS imaging was demonstrated to corroborate synchrotron radiation confocal X-ray fluorescence (SR-XRF) microimaging of single cells investigated under in vivo conditions.

[Construction of NIRS-based total quality control system for compound Ejiao oral liquid and relative thinking].

In this paper, near infrared spectroscopy (NIRS)-based total quality control system of compound Ejiao oral liquid is introduced briefly, including the quality control of raw traditional Chinese medicine (TCM) materials, monitoring and control of the extract and the alkaline precipitation technics, and also the inspection of finished products in both open bottle and non-opening modes. By analyzing and summing up the significance and difficulties, several important problems in the practical applications of NIRS technology are proposed, which will provide references for the similar studies of other TCM products.

Application of QbD principles for the evaluation of empty hard capsules as an input parameter in formulation development and manufacturing.

Understanding the product and process variable on the final product performance is an essential part of the quality-by-design (QbD) principles in pharmaceutical development. The hard capsule is an established pharmaceutical dosage form used worldwide in development and manufacturing. The empty hard capsules are supplied as an excipient that is filled by pharmaceutical manufacturers with a variety of different formulations and products. To understand the potential variations of the empty hard capsules as an input parameter and its potential impact on the finished product quality, a study was performed investigating the critical quality parameters within and in between different batches of empty hard gelatin capsules. The variability of the hard capsules showed high consistency within the specification of the critical quality parameters. This also accounts for the disintegration times, when automatic endpoint detection was used. Based on these data, hard capsules can be considered as a suitable excipient for product development using QbD principles.

Conventional and real-time PCR-based approaches for molecular detection and quantitation of bovine species material in edible gelatin.

The majority of edible gelatin in Europe is derived from pigskin, but a significant portion is extracted from bovine tissue. Because of the bovine spongiform encephalopathy crisis, consumers might be concerned about the gelatin used in various products. To assure consumers of the quality and safety of edible gelatin, European Union directive 1999/724/EC described general guidelines for gelatin production, including requirements for documentary proof confirming that raw materials are from animals fit for human consumption. Analytical methods to confirm gelatin documentation or raw material animal species source in the finished product are lacking. In this study, several published species-specific PCR systems were evaluated as potential molecular methods for determining the origin of the raw material used in making gelatin. A recently validated bovine species-specific PCR primer set targeting the ATPase 8 subunit gene in bovine mitochondrial DNA was suitable for detection of bovine material in gelatin. This PCR primer set was optimized using conventional and real-time PCR approaches. An evaluation of these two PCR methods confirmed the high specificity for the adopted primer set in various gelatin matrices of known origin. The inclusion of bovine gelatin in pork or fish gelatin can be detected at 0.1 to 0.001%. These PCR assays are potential molecular detection tools that can be used to routinely detect bovine gelatin either alone or as an inclusion in gelatin made from other species.

Enteric coating of soft gelatin capsules by spouted bed: effect of operating conditions on coating efficiency and on product quality.

The present study was conducted in order to analyze the viability of the spouted bed process for application of a gastric-resistant coating to soft gelatin capsules. The variables investigated were: included angle of conical base, (gamma), the relation between the feed mass flow rate of the coating suspension and the feed mass flow rate of spouting gas (W(s)/W(g)); the ratio between the flow rate of the spouting gas and the flow rate at minimum spouting condition (Q/Q(ms)); the mass of capsules in the bed (M(0)), and the capsule's size. The product quality was measured by disintegration tests, traction x deformation tests, image analysis and by the evaluation of the coating mass distribution and shape factor variation during the coating operation. The experiments were performed in a spouted bed with a column diameter of 200 mm and included a conical base angle of 40 degrees. The best coating efficiency values were obtained for M(0)=300 g. Coating efficiency tended to increase with increasing W(s)/W(g) ratio. Disintegration tests showed that the gastric-resistant effect was obtained with a coating mass of 3.86 mg/cm(2). The shape factor increase during the coating operation. The capsule's coating mass distribution tended to maintain the original distribution.

[The estimation of the effectiveness of GRF glue in the respiratory].

We had reported on the basis of experimental findings that the efficacy of protecting the suture line in tracheoplasty by using a self-fascia lata and GRF glue. This time we investigated the effectiveness of GRF glue in the respiratory surgery on the basis of clinical findings. All ten cases in which GRF glue was used resulted in an excellent outcome, namely, GRF glue prevented from air leakage, bleeding and leakage of chyle. Moreover, it was not recognized that any grave side effects occurred in any cases after using GRF glue. We could conclude that GRF glue was a useful material for the respiratory surgery, especially for the case that had a dead space after lobectomy. However its price is not inexpensive, therefore, there seems need to restrict the use of GRF glue--for example--to cases of Giant bulla resection, residual of a large dead space as a postoperative possibility and chemotherapy performed before an operation, and so on.

Influence of gelatin and bovine serum lubricants on ultra-high molecular weight polyethylene wear debris generated in in vitro simulations.

Ultra-high molecular weight polyethylene (UHMWPE) wear debris induced osteolysis has a major role in the late aseptic loosening and ultimate failure of total hip replacements (THR). Clinically relevant in vitro simulations of wear are essential to predict the osteolytic potential of bearing surfaces in artificial hip joints. Newborn calf or bovine serum has been accepted as a boundary lubricant for such in vitro tests, but its biological stability has been questioned. This study compared the wear factors, number of wear particles and levels of microbial contamination produced in bovine serum and a gelatin-based lubricant. The wear factors produced by the two lubricants were not significantly different, however the wear debris morphology produced was substantially different. The bovine serum became contaminated with micro-organisms within 28 h, whereas the protein-based lubricant remained uncontaminated. The results showed that bovine serum was not a stable boundary lubricant. They also showed that although the wear factors for the two solutions were not significantly different, the protein-based lubricant was not a suitable alternative to bovine serum because the wear debris produced was not clinically relevant.

Injectable soft tissue substitutes.

Historical and modern advances in the development of an injectable soft tissue substitute are reviewed. Nonbiologic alloplastic and biologic injectables are described. The authors' experiences, as well as those of others, employing presently available materials in terms of specific indications and special techniques are delineated. The search for a safe, effective, easy-to-use, and long-lasting soft tissue substitute continues.

Use of beef extract as a diluent in direct agglutination test for visceral leishmaniasis.

Direct Agglutination Tests (DAT) have been developed for sero-diagnosis of visceral leishmaniasis (VL). The objective of this study was to find a more suitable diluent for serum dilution in a Direct Agglutination Test. 1% of beef extract (BEX) was used as a diluent in the Direct Agglutination Test; 1% foetal bovine serum (FBS) and 0.2% gelatine (Gel) were included for comparison as alternative diluents. Serum from VL patients, individuals suffering from hydatidosis, syphilis and from healthy adult Kenyans was used as control. Beef extract was found to be easy to handle, less expensive and results comparable to those obtained from the other diluents.

Urostomy appliances and stoma care routines. The relation to peristomal skin complications.

The aim of the study was to describe types of appliances and stoma care routines and evaluate their relation to peristomal skin complications. Sixty-six patients with a cutaneous uretero-ileostomy were interviewed and the peristomal skin was assessed according to Classification of Peristomal Skin (CPS). The results show a conservatism regarding the types of appliance and the stoma care routines. More than half of the patients used the same product at follow-up as they were initially fitted with three to 14 years earlier. The routines adopted by the patients were often inadequate, resulting in skin complications. Continuous exposure of the skin to urine by creation of a too wide opening in the face-plate and infrequent changing of the appliance resulted in development of pseudoverrucose skin lesions.

Biomechanical optimization of a model particulate composite for orthopaedic applications.

Particulate composites are a potential solution to the need for an injectable, biocompatible, resorbable material that could be used to reinforce fractures and defects in bone and temporarily to stabilize porous ingrowth prostheses. We have developed a model system for producing and testing particulate composites to determine if mechanical properties suitable for orthopaedic applications can be achieved. The experiments used bovine cortical bone and various forms of hydroxyapatite for the particulate phase and a collagen and particulate reinforce gelatin-resorcinol-formaldehyde (G-R-F) adhesive for the matrix phase. Using unconfined compression testing, we measured the effects of variation in particulate type, size, shape, and volume fraction on the material properties of the particulate composites. We found that compressive strengths greater than 10 MPa and compressive moduli greater than 100 MPa could be achieved in this model system. Rough and irregular particulates exhibited higher compressive strengths and moduli than smooth and spherical particulates. Mechanical properties were largely independent of particulate size in the range of 125-850 microns diameter. This model system suggests that, with the development of new biocompatible matrix materials, particulate composites with mechanical properties suitable for orthopaedic applications can be achieved.

Silastic and gelatin film sheeting in tympanoplasty.

In 78 patients, representing 10% of the most severe case of chronic otitis media and in whom the mucosa from the promontory had been removed or severely damaged, the promontory was covered in one-stage mastoidectomy-tympanoplasty with a large piece of thin silastic or of Gelfilm. The results in the silastic group, comprising 50 patients, and in the Gelfilm group, comprising 28 patients, were compared on the basis of various criteria. The results proved better in the Gelfilm group. In particular, the percentage of recurrent perforations was considerably lower than in the silastic group. In one-stage operations, which we prefer, it is therefore recommended to use Gelfilm rather that silastic.