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Protein hydrolysates are an important part of the human diet. Often, they are prepared from milk, soy, or collagen. In the present study, four different collagen hydrolysates were tested, varying in the average molecular weight and the animal source. Three types of samples, the dissolved start products, in vitro generated dialysates (containing the digested components that are potentially available for small intestinal absorption), and human serum collected after product ingestion, were analyzed using LC-MS to compare the state of the hydrolysates before and after absorption, i.e., uptake into the blood. It was found that the composition of the collagen hydrolysates prior to and after ingestion was highly complex and dynamic, which made it challenging to predefine a strategy for a targeted analysis. Therefore, we implemented a new analytical approach to first map hydrolysate data sets by performing non-targeted LC-MS analysis followed by non-targeted and targeted data analysis. It was shown that the insight gained by following such a top down (data) analytical workflow could be crucial for defining a suitable targeted setup and considering data trends beyond the defined targets. After having defined and performed a limited targeted analysis, it was found that, in our experimental setup, Hyp-Gly and especially Pro-Hyp contributed significantly as carrier to the total Hyp increase in blood after ingestion of collagen hydrolysate.

Terahertz and millimeter waves penetrate various dielectric materials, including plastics, ceramics, crystals, and concrete, allowing terahertz transmission and reflection images to be considered as a new imaging tool complementary to X-Ray or Infrared. Terahertz imaging is a well-established technique in various laboratory and industrial applications. However, these images are often two-dimensional. Three-dimensional, transmission-mode imaging is limited to thin samples, due to the absorption of the sample accumulated in the propagation direction. A tomographic imaging procedure can be used to acquire and to render three-dimensional images in the terahertz frequency range, as in the optical, infrared or X-ray regions of the electromagnetic spectrum. In this paper, after a brief introduction to two dimensional millimeter waves and terahertz imaging we establish the principles of tomography for Terahertz Computed tomography (CT), tomosynthesis (TS), synthetic aperture radar (SAR) and time-of-flight (TOF) terahertz tomography. For each technique, we present advantages, drawbacks and limitations for imaging the internal structure of an object.

It can be important for consumers to know whether food products contain animal material and, if so, of which species. Food products with animal material as an ingredient often contain collagen type 1. LC-MS/MS (Liquid Chromatography–tandem Mass Spectrometry) was applied as technique to generically detect bird. Unlike for example fish, that have experienced longer divergence times, it is still possible to find generic LC-MS targets for avian type 1 collagen. After theoretical target selection using 83 collagen 1α2 bird sequences of 33 orders and construction of a common ancestor sequence of birds, experimental evidence was provided by analyzing extracts from 10 extant bird species. Two suitable options have been identified. The combination of VGPIGPAGNR and VGPIGAAGNR (pheasant only) covers all investigated birds and was not found in other species. The peptide EGPVGF p GADGR covers all investigated birds, but also occurs in several species of crocodiles and turtles. The presence of the generic peptide (combination) was confirmed in food products, proving the principle, and can therefore be used to detect the presence of bird. Furthermore, it is shown how the use of constructed ancestor sequences could benefit the field of paleoproteomics, in the interpretation of collagen MS/MS spectra of ancient species. Our theoretical analysis and assessment of reported Brachylophosaurus canadensis collagen 1α2 MS/MS data provided support for several previous peptide sequence assignments, but we also propose that our constructed ancestral bird sequence GP p GESGAVGPAGPIGSR may fit the MS/MS data better than the original assignment GLPGESGAVGPAGP p GSR.

Collagen is an important structural protein and the most abundant protein in mammals. In several research fields, structural analysis of collagens is performed. Fibrillar collagens almost entirely consist of continuous repeats of GXY, where G is glycine, X is often proline or alanine and Y is often hydroxyproline or alanine. In the present study, the collagen structure was investigated in detail at the nucleotide, codon group, amino acid and target peptide level using sequence analyses. One of the most important findings was that a selection of codon groups is predominantly involved in amino acid changes between closely related collagens and that other change routes come up when collagens are less related. The findings of the sequence analyses were used to evaluate reported sequences of non-avian dinosaur species and database entries of duck and chicken collagen. The duck assessment was supported by an experimental data set, obtained by collagen extraction from duck skin and subsequent digestion and LC–MS analysis. It was found that database entries of chicken and duck collagen 3α1 contained unreliable features, such as missing parts, no continuous GXY pattern and too many interspecies differences. As an example, the erroneous nature of one of these unreliable features was confirmed experimentally using LC–MS. Finally, dino and bird collagen 1α1 were compared. The presented results will show that performing a domain-specific proteogenomic analysis provides very useful information to assess de novo sequencing results and database information of collagens. Furthermore, it offers deeper insight in the functional restrictions and routes of evolutionary divergence.

Hemolysis can result in analyte suppression or enhancement and it can affect the extraction efficiency and analyte stability. Triskelion developed an LC–MS method to monitor hemolysis. The concept can be integrated into existing and new quantitative protein LC–MS methods and can be validated according to the most appropriate tier. In this proof of concept study, the tryptic target LLVVYPWTQR was used to quantify hemoglobin. The peptide target has only few variations considering the most common (laboratory) animals and is thus nearly generic. It was shown that LC–MS is a suitable technique for the quantification of hemoglobin in hemolyzed samples and that the signals are not affected by lipemia. LC–MS exhibited the best performance to monitor hemolysis when the results were compared with UV–VIS and visual inspection, especially when samples were lipemic.

Background: In laryngectomized patients, tracheoesophageal voice generally provides a better voice quality than esophageal voice. Understanding the aerodynamics of voice production in patients with a voice prosthesis is important for optimizing prosthetic designs and successful voice rehabilitation. Objectives: To measure the aerodynamics and sound intensity in tracheoesophageal voice production. Study Design and Methods: We built a special setup, which consisted of a Pentium 200 MHz computer with an AD-DA interface card and Labview 4.01 software. In an oral/nasal mask we constructed several mass flow sensors and a microphone. This measured both the oral airflow and the level of sound. For the measurement of endotracheal pressure, which is the driving force behind the airflow, we used a transducer which was connected to the tracheostoma. The endoesophageal pressure was measured at the level of the prosthesis in the esophagus by a Mikrotip transducer. Using this we could determine how much the voice prosthesis contributes to the overall pressure drop of the phonatory tract. Furthermore, the average airflow rate as a function of the sound pressure levels could be determined. Results: In our population, 6 out of 7 patients showed a positive relationship between trans-source airflow and generated sound intensity. We compared our prosthesis pressure drop values with in vitro data and found that there are some differences, possibly due to difference in age of the prosthesis and physiological circumstances in vivo. The overall contribution of the voice prosthesis to the airway resistance depends on the level of phonation and the type of device. In our patient group it is apparent that the pharyngoesophageal (PE) segment has the greatest share of the total pressure drop, especially at higher airflow rates. We measured a 27% pressure drop in airflow over the voice prosthesis. Different tracheostoma occlusion methods did not have any effect on the aerodynamics and sound intensity. One patient that had had a jejunal graft for reconstruction showed, not unexpectedly, extremely different aerodynamic values. We were unable to define optimal airflow rates or optimal resistance values for sound production in the PE segment. Conclusion: The aerodynamic characteristics of voice production in laryngectomized patients with voice prostheses are determined by both prosthetic factors and PE segment tissue factors. In our patient group the PE segment is responsible for the greatest pressure drop. We found no significant difference in pressure drop and sound intensity for different tracheostoma occlusion methods.

Mycosporine-like amino acids (MAAs) are UVR-absorbing compounds ubiquitous in the aquatic environment. Metazoans appear to obtain them from their diet or from symbiotic microorganisms. The study of MAAs has been complicated by the lack of commercially available standards. Use of tandem mass spectrometry renders this problem tractable. My dissertation exploited the availability of previously published MAA fragmentation patterns to identify MAAs in the mucus of the corallivorous territorial butterflyfish, Chaetodon multicinctus. I next adapted both a powerful HPLC separation procedure and a tandem mass spectrometry technique to not only identify, but also quantify, MAAs in the absence of standards. With this method I examined MAAs through the trophic chain in C. multicinctus and its coral prey, Pocillopora meandrina, Porites compressa and Porites lobata. Specimens from three depths provided non-overlapping levels of downwelling UVR irradiance to investigate if the four species studied modulated their MAA content with exposure level. I found that the MAAs present in Chaetodon multicinctus epithelial mucus are different from the MAAs in their diet: some, like mycosporine-glycine, abundant in the coral diet, are not detectable in fish mucus and some, present in the epidermal mucus such as palythene and usujirene, are not found in the diet. These results are consistent with prior research on fish eyes, sea urchins, and pteropods. They suggest the possibility of selective uptake, translocation and transformation of MAAs by metazoan consumer organisms. The details of these processes are unknown. MAA concentrations in the three coral species decreased with increasing depth and the associated UVR reduction. This pattern was, however, not observed in the fish probably due to a ceiling effect. A model is proposed to illustrate this possibility. The corals also showed an interesting difference in MAAs diversity perhaps due to differences in their Symbiodinium: Porites spp. known to have low diversity of symbionts also had a low diversity of MAAs compared to Pocillopora meandrina that harbors a greater diversity of zooxanthellae. My work also resulted in the addition of several MS2 patterns useful for identification. Further, the discovery of up to four new MAAs suggests the value of continuing to investigate these intriguing metabolites.