Effects of conditioning temperature and time during the pelleting process on feed molecular structure, pellet durability index, and metabolic features of co-products from bio-oil processing in dairy cows.

The objectives of this study were to systematically determine effects of conditioning temperature (70, 80, and 90°C), time (50 and 75 s), and interaction (temperature × time) during the pelleting process on co-products from bio-oil processing (canola meal) in terms of processing-induced changes on (1) protein molecular structure, (2) pellet durability index, (3) detailed chemical profile, (4) metabolic features and fractions of protein and carbohydrate, (5) total digestible nutrients and energy values, and (6) rumen degradable and undegradable content. Pellet durability was increased with increasing conditioning time. Chemical and carbohydrate profiles of co-products were not altered by pelleting process under different conditioning temperatures and times. With regard to protein fraction profiles, pellets conditioned for 50 s had higher soluble crude protein (SCP) and lower neutral detergent insoluble crude protein (NDICP) contents than those conditioned for 75 s (21.7 vs. 20.1% SCP, 16.0 vs. 16.5% NDICP, respectively). Total digestible nutrients and energy values were not altered by processing. Samples conditioned for 50 s had a higher content of rapidly degradable protein fraction (PA2) than those conditioned for 75 s (21.7 vs. 21.1% crude protein). In addition, the slowly degradable true protein fraction (PB2) was affected by the interaction of conditioning temperature and time. However, carbohydrate fractions did not differ with different conditioning temperatures and time. Different temperatures and time of conditioning during pelleting process greatly affect protein profiles without altering carbohydrate profiles. Molecular structure analyses also showed that pelleting altered inherent protein molecular structures of the co-products from bio-oil processing. Future study is needed to detect how molecular structure changes affect nutrient availability in dairy cattle.

Introduction of soft X-ray spectromicroscopy as an advanced technique for plant biopolymers research.

Soft X-ray absorption spectroscopy coupled with nano-scale microscopy has been widely used in material science, environmental science, and physical sciences. In this work, the advantages of soft X-ray absorption spectromicroscopy for plant biopolymer research were demonstrated by determining the chemical sensitivity of the technique to identify common plant biopolymers and to map the distributions of biopolymers in plant samples. The chemical sensitivity of soft X-ray spectroscopy to study biopolymers was determined by recording the spectra of common plant biopolymers using soft X-ray and Fourier Transform mid Infrared (FT-IR) spectroscopy techniques. The soft X-ray spectra of lignin, cellulose, and polygalacturonic acid have distinct spectral features. However, there were no distinct differences between cellulose and hemicellulose spectra. Mid infrared spectra of all biopolymers were unique and there were differences between the spectra of water soluble and insoluble xylans. The advantage of nano-scale spatial resolution exploited using soft X-ray spectromicroscopy for plant biopolymer research was demonstrated by mapping plant cell wall biopolymers in a lentil stem section and compared with the FT-IR spectromicroscopy data from the same sample. The soft X-ray spectromicroscopy enables mapping of biopolymers at the sub-cellular (~30 nm) resolution whereas, the limited spatial resolution in the micron scale range in the FT-IR spectromicroscopy made it difficult to identify the localized distribution of biopolymers. The advantages and limitations of soft X-ray and FT-IR spectromicroscopy techniques for biopolymer research are also discussed.

Detection of protein structure of frozen ancient human remains recovered from a glacier in Canada using synchrotron fourier transform infrared microspectroscopy.

We previously used synchrotron infrared microspectroscopy to describe the biochemical signature of skeletal muscle (biceps brachii) from the frozen ancient remains of a young man. In this current paper, we use light microscopy to assess the state of preservation of cellular components in the trapezius muscle from these same ancient remains and then use mid-infrared analysis at the Canadian Light Source synchrotron facility to further analyze the tissue. We compare spectra between the trapezius samples from the ancient remains and a recently deceased cadaver (control). Infrared spectra indicate preservation of secondary structure, with the α-helix being the principal component, along with triple helical portions of the protein backbone. Our mid-infrared analysis indicates an energy reserve in the skeletal muscle in the ancient remains.

Use of a dry fractionation process to manipulate the chemical profile and nutrient supply of a coproduct from bioethanol processing.

With an available processing technology (fractionation), coproducts from bioethanol processing (wheat dried distillers grains with solubles, DDGS) could be fractionated to a desired/optimal chemical and nutrient profile. There is no study, to the author's knowledge, on manipulating nutrient profiles through fractionation processing in bioethanol coproducts in ruminants. The objectives of this study were to investigate the effect of fractionation processing of a coproduct from bioethanol processing (wheat DDGS) on the metabolic characteristics of the proteins and to study the effects of fractionation processing on the magnitude of changes in chemical and nutrient supply to ruminants by comparing chemical and nutrient characterization, in situ rumen degradation kinetics, truly absorbed protein supply, and protein degraded balance among different fractions of coproduct of wheat DDGS. In this study, wheat DDGS was dry fractionationed into A, B, C, and D fractions according to particle size, gravity, and protein and fiber contents. The results showed that the fractionation processing changed wheat DDGS chemical and nutrient profiles. NDF and ADF increased from fraction A to D (NDF, from 330 to 424; ADF, from 135 to 175 g/kg DM). Subsequently, CP decreased (CP, from 499 to 363 g/kg DM), whereas soluble CP, NPN, and carbohydrate increased (SCP, from 247 to 304 g/kg CP; NPN, from 476 to 943 g/kg SCP; CHO, from 409 to 538 g/kg DM) from fraction A to D. The CNCPS protein and carbohydrate subfractions were also changed by the fractionation processing. Effective degradability of DM and CP and total digestible protein decreased from fraction A to D (EDDM, from 734 to 649; EDCP, from 321 to 241; TDP, from 442 to 312 g/kg DM). Total truly absorbed protein in the small intestine decreased from fraction A to D (DVE value, from 186 to 124 g/kg DM; MP in NRC-2001, from 193 to 136 g/kg DM). Degraded protein balance decreased from wheat DDGS fractions A-D (DPB in the DVE/OEB system, from 245 to 161 g/kg DM; DPB in NRC-2001, from 242 to 158 g/kg DM). The fractionation processing had a great impact on the chemical and nutrition profiles. Total truly digested and absorbed protein supply and degraded protein balance were decreased. The processing relatively optimized the protein degraded balance of the coproducts to dairy cattle. Compared with the original wheat DDGS (without fractionation), fractionation processing decreased truly absorbed protein supply of DVE and MP values. In conclusion, fractionation processing can be used to manipulate the nutrient supply and N-to-energy degradation synchronization ratio of coproducts from bioethanol processing. Among the fractions, fraction A was the best in terms of its highest truly absorbed protein DVE and MP values. Fractionation processing has great potential to fractionate a coproduct into a desired and optimal chemical and nutrient profile. To the author's knowledge, this is the first paper to show that with fractionation processing, the coproducts from bioethanol processing (wheat DDGS) could be manipulated to provide a desired/optimized nutrient supply to ruminants.

A new approach to concordance in mid-infrared spectromicroscopy mapping of malignant tumors.

Mid-infrared spectromicroscopy studies on biological tissue sections require accurate identification of tumor-bearing areas in histology-stained and infrared-unstained tissue sections. Concordance was achieved as follows: paired stained and unstained thin (5 microm) human brain tumor cryosections mounted on slides were scanned with a Nikon Coolscan 4000 film scanner at 4000 dpi, edited with Adobe Photoshop CS2 software, and both digital images saved. A digital tractile grid, developed in our laboratory, was overlaid onto both images. Boundaries of tumor-containing areas in stained sections were identified by light microscopy, and a digital boundary map constructed. The map was transferred onto the unstained spectromicroscopy tissue image, and finally layered onto the gridded, equisized, spectromicroscope-generated overview image prior to Fourier transform infrared spectromicroscopy. Accurate identification of tumor-bearing areas, normal brain tissue and transitional zones allowed for meaningful interpretation of respective spectral patterns in detecting subtle differences within biochemical profiles. This is the first reported method of a standardized technique for ensuring concordance in mapping of malignant tumors by mid-infrared spectromicroscopy. This technique is applicable to all biological thin tissue sections, and serves to enhance accuracy of concordance between globar- and synchrotron-light generated infrared data with that obtained by conventional light microscopy.

Fourier transform infrared microspectroscopic analysis of the effects of cereal type and variety within a type of grain on structural makeup in relation to rumen degradation kinetics.

The objectives of this study were to use Fourier transform infrared microspectroscopy (FTIRM) to determine structural makeup (features) of cereal grain endosperm tissue and to reveal and identify differences in protein and carbohydrate structural makeup between different cereal types (corn vs barley) and between different varieties within a grain (barley CDC Bold, CDC Dolly, Harrington, and Valier). Another objective was to investigate how these structural features relate to rumen degradation kinetics. The items assessed included (1) structural differences in protein amide I to nonstructural carbohydrate (NSC, starch) intensity and ratio within cellular dimensions; (2) molecular structural differences in the secondary structure profile of protein, alpha-helix, beta-sheet, and their ratio; (3) structural differences in NSC to amide I ratio profile. From the results, it was observed that (1) comparison between grain types [corn (cv. Pioneer 39P78) vs barley (cv. Harrington)] showed significant differences in structural makeup in terms of NSC, amide I to NSC ratio, and rumen degradation kinetics (degradation ratio, effective degradability of dry matter, protein and NSC) (P < 0.05); (2) comparison between varieties within a grain (barley varieties) also showed significant differences in structural makeup in terms of amide I, NSC, amide I to NSC ratio, alpha-helix and beta-sheet protein structures, and rumen degradation kinetics (effective degradability of dry matter, protein, and NSC) (P < 0.05); (3) correlation analysis showed that the amide I to NSC ratio was strongly correlated with rumen degradation kinetics in terms of the degradation rate (R = 0.91, P = 0.086) and effective degradability of dry matter (R = 0.93, P = 0.071). The results suggest that with the FTIRM technique, the structural makeup differences between cereal types and between different varieties within a type of grain could be revealed. These structural makeup differences were related to the rate and extent of rumen degradation.

Fourier transform infrared spectromicroscopy and hierarchical cluster analysis of human meningiomas.

Limitations of conventional light microscopy in pathological diagnosis of brain tumors include subjective bias in interpretation and discordance of nomenclature. A study using mid-infrared (IR) spectromicroscopy was undertaken to determine whether meningiomas, a group of brain tumors prone to recurrence, could be identified by the unique spectral 'fingerprints' of their chemical composition. Paired, thin (5-microm) cryosections of snap-frozen human meningioma tumor samples removed at elective surgery were mounted on glass (hematoxylin and eosin-stained tissue section) and infrared (unstained tissue section) reflectance slides, respectively. Concordance of the tumor-bearing areas identified in the stained section by a pathologist with the unstained IR tissue section was ensured using a novel digital grid and tumor-mapping system developed in our laboratory. Compared with the normal control, tumor samples from four meningioma patients revealed a marked decrease in bands associated with unsaturated fatty acids, particularly in the bands at 3010, 2920, 2850, and 1735 cm(-1). Spectral datasets were subjected to hierarchical cluster analyses (HCA) using Ward's algorithm for comparison and grouping of similar data groups, and were converted into color-coded digital maps for matching spectra with their respective clusters. False color images of 5 and 6 clusters obtained by HCA identified dominant clusters corresponding to tumor tissue. Corroboration of these findings in a larger number of meningiomas may allow for more precise identification of these and other types of brain tumors.

Inferring the geometry of fourth-period metallic elements in arabidopsis thaliana seeds using synchrotron-based multi-angle X-ray fluorescence mapping.

BACKGROUND: Improving our knowledge of plant metal metabolism is facilitated by the use of analytical techniques to map the distribution of elements in tissues. One such technique is X-ray fluorescence (XRF), which has been used previously to map metal distribution in both two and three dimensions. One of the difficulties of mapping metal distribution in two dimensions is that it can be difficult to normalize for tissue thickness. When mapping metal distribution in three dimensions, the time required to collect the data can become a major constraint. In this article a compromise is suggested between two- and three-dimensional mapping using multi-angle XRF imaging. METHODS: A synchrotron-based XRF microprobe was used to map the distribution of K, Ca, Mn, Fe, Ni, Cu and Zn in whole Arabidopsis thaliana seeds. Relative concentrations of each element were determined by measuring fluorescence emitted from a 10 microm excitation beam at 13 keV. XRF spectra were collected from an array of points with 25 or 30 microm steps. Maps were recorded at 0 and 90 degrees , or at 0, 60 and 120 degrees for each seed. Using these data, circular or ellipsoidal cross-sections were modelled, and from these an apparent pathlength for the excitation beam was calculated to normalize the data. Elemental distribution was mapped in seeds from ecotype Columbia-4 plants, as well as the metal accumulation mutants manganese accumulator 1 (man1) and nicotianamine synthetase (nasx). CONCLUSIONS: Multi-angle XRF imaging will be useful for mapping elemental distribution in plant tissues. It offers a compromise between two- and three-dimensional XRF mapping, as far as collection times, image resolution and ease of visualization. It is also complementary to other metal-mapping techniques. Mn, Fe and Cu had tissue-specific accumulation patterns. Metal accumulation patterns were different between seeds of the Col-4, man1 and nasx genotypes.

Imaging molecular chemistry of Pioneer corn.

Synchrotron Fourier transform infrared (FTIR) microspectroscopy as a rapid, direct, and nondestructive analytical technique can explore molecular chemical features of the microstructure of biological samples. The objective of this study was to use synchrotron FTIR microspectroscopy to image the molecular chemistry of corn (cv. Pioneer 39P78) to reveal spatial intensity and distribution of chemical functional groups in corn tissue. This experiment was performed at the U2B station of the National Synchrotron Light Source in Brookhaven National Laboratory (NSLS-BNL, Upton, NY). The Pioneer corn tissue was imaged from the pericarp, seed coat, aleurone, and endosperm under peaks at 1736 (carbonyl C=O ester), 1510 (aromatic compound), 1650 (amide I), 1550 (amide II), 1246 (cellulosic material), 1160 (CHO), 1150 (CHO), 1080 (CHO), 929 (CHO), 860 (CHO), 3350 (OH and NH stretching), 2929 (CH(2) stretching band), and 2885 cm(-1) (CH(3) stretching band). The results showed that with synchrotron FTIR microspectroscopy, the images of the molecular chemistry of Pioneer corn could be generated. Such information on the microstructural-chemical features of grain corn can also be used for corn breeding programs for selecting superior varieties of corn for targeted food and feed purposes and for prediction of corn quality and nutritive value for humans and animals.

Using synchrotron-based FTIR microspectroscopy to reveal chemical features of feather protein secondary structure: comparison with other feed protein sources.

Studying the secondary structure of proteins leads to an understanding of the components that make up a whole protein. An understanding of the structure of the whole protein is often vital to understanding its digestive behavior in animals and nutritive quality. Usually protein secondary structures include alpha-helix and beta-sheet. The percentages of these two structures in protein secondary structures may influence feed protein quality and digestive behavior. Feathers are widely available as a potential protein supplement. They are very high in protein (84%), but the digestibility of the protein is very low (5%). The objective of this study was to use synchrotron-based Fourier transform infrared (FTIR) microspectroscopy to reveal chemical features of feather protein secondary structure within amide I at ultraspatial resolution (pixel size = 10 x 10 microm), in comparison with other protein sources from easily digested feeds such as barley, oat, and wheat tissue at endosperm regions (without destruction of their inherent structure). This experiment was performed at beamline U2B of the Albert Einstein Center for Synchrotron Biosciences at the National Synchrotron Light Source (NSLS) in Brookhaven National Laboratory (BNL), U.S. Dept of Energy (NSLS-BNL, Upton, NY). The results showed that ultraspatially resolved chemical imaging of feed protein secondary structure in terms of beta-sheet to alpha-helix peak height ratio by stepping in pixel-sized increments was obtained. Using synchrotron FTIR microspectroscopy can distinguish structures of protein amide I among the different feed protein sources. The results show that the secondary structure of feather protein differed from those of other feed protein sources in terms of the line-shape and position of amide I. The feather protein amide I peaked at approximately 1630 cm(-1). However, other feed protein sources showed a peak at approximately 1650 cm(-1). By using multicomponent peak modeling, the relatively quantitative amounts of alpha-helix and beta-sheet in protein secondary structure were obtained, which showed that feather contains 88% beta-sheet and 4% alpha-helix, barley contains 17% beta-sheet and 71% alpha-helix, oat contains 2% beta-sheet and 92% alpha-helix, and wheat contains 42% beta-sheet and 50% alpha-helix. The difference in percentage of protein secondary structure may be part of the reason for different feed protein digestive behaviors. These results demonstrate the potential of highly spatially resolved infrared microspectroscopy to reveal feed protein secondary structure. Information from this study by the infrared probing of feed protein secondary structure may be valuable as a guide for feed breeders to improve and maintain protein quality for animal use.

Using synchrotron transmission FTIR microspectroscopy as a rapid, direct, and nondestructive analytical technique to reveal molecular microstructural-chemical features within tissue in grain barley.

The objective of this study was to use synchrotron transmission FTIR microspectroscopy as a rapid, direct, and nondestructive analytical technique to reveal molecular microstructural-chemical features within tissue in grain barley. The results showed that synchrotron transmission FTIR microspectroscopy could provide spectral, chemical, and functional group characteristics of grain barley tissue at ultrahigh spatial resolutions. The spatially localized structural-chemical distributions of biological components (lignin, cellulose, protein, lipid, and carbohydrates) and biological component ratios could be imaged. Such information on molecular microstructural-chemical features within the tissue can be used for plant breeding programs for selecting superior varieties of barley for special purposes and for prediction of grain barley quality and nutritive value for humans and animals.

Chemical imaging of microstructures of plant tissues within cellular dimension using synchrotron infrared microspectroscopy.

Synchrotron radiation-based Fourier transform infrared microspectroscopy (SR-FTIR) is an advanced bioanalytical technique capable of exploring the chemistry within microstructures of plant and animal tissues with a high signal to noise ratio at high ultraspatial resolutions (3-10 microm) without destruction of the intrinsic structures of a tissue. This technique is able to provide information relating to the quantity, composition, structure, and distribution of chemical constituents and functional groups in a tissue. The objective of this study was to illustrate how the SR-FTIR technique can be used to image inherent structures of plant tissues on a cellular level (pixel size, approximately 10 microm x 10 microm). The results showed that with the extremely bright synchrotron light, spectra with high signal to noise ratios were obtained from areas as small as 10 microm x 10 microm in the plant tissue, which allowed us to "see" plant tissue in a chemical sense on a cellular level. The ultraspatial resolved imaging of plant tissues by stepping in pixel-sized increments was obtained. Chemical distributions of plant tissues such as lignin, cellulose, protein, lipid, and total carbohydrate could be mapped. These images revealed the chemical information of plant intrinsic structure. In conclusion, SR-FTIR can provide chemical and functional characteristics of plant tissue at high ultraspatial resolutions. The SR-FTIR microspectroscopic images can generate spatially localized functional group and chemical information within cellular dimensions.