RESUMEN
Fasting of up to 24 hr has been shown to increase intestinal permeability (IP) in chickens. The aim of this study was to determine whether fasting duration of 4.5 and 9 hr increased IP and whether l-glutamine (a non-essential amino acid) supplementation before fasting provided some protection of barrier function as shown in other species. Ross 308 male broilers (n = 96) were fed either a control diet or the same diet supplemented with 1% glutamine from d0 to d38 post-hatch. On d37, the birds were assigned to single-bird metabolism cages and were fasted for either 0, 4.5, 9 or 19.5 hr. This study design was 2 × 4 factorial with two levels of glutamine and four levels of fasting. Birds in the 0-hr fasting group had free access to feed. All birds had ad libitum access to water. To measure IP on day 38, following their respective fasting periods, birds were administered two separate oral gavages of fluorescein isothiocyanate dextran (FITC-d) followed by lactulose, mannitol and rhamnose (LMR) sugars, 60 min apart. Whole blood was collected from the jugular vein 90 min post-LMR sugar gavage. FITC-d and L/M/R ratios were measured by spectrophotometry and high-performance ionic chromatography respectively. Lipopolysaccharide (LPS) endotoxins in plasma of the birds fed the control diet were also measured using chicken-specific LPS antibody ELISA. Serum FITC-d and plasma L/M and L/R ratios for 4.5, 9 and 19.5 hr were significantly (p < .05) higher compared to the non-fasting group. However, IP was not different in the glutamine-supplemented group (p > .05) compared to the control group. LPS concentrations measured by the ELISA were below the detectable range. We conclude that fasting periods of 4.5 and 9 hr increased IP compared to non-fasted birds and dietary glutamine supplementation did not ameliorate changes in IP.
Asunto(s)
Pollos/fisiología , Privación de Alimentos , Alimentación Animal/análisis , Animales , Dextranos , Dieta/veterinaria , Fluoresceína-5-Isotiocianato/análogos & derivados , Glutamina , Intestinos , Lactulosa/sangre , Masculino , Manitol/sangre , Permeabilidad , Ramnosa/sangre , Factores de TiempoRESUMEN
Short-term fasting for 4.5 and 9 hr has been demonstrated to increase intestinal permeability (IP) in chickens. This study aimed to investigate the effects of 0, 4.5, 9 and 19.5 hr fasting on intestinal gene expression and villus-crypt architecture of enterocytes in jejunal and ileal samples. On day 38, Ross-308 male birds were fasted according to their group and then euthanised. Two separate intestinal sections (each 2 cm long, jejunum and ileum) were collected. One section was utilised for villus height and crypt depth measurements. The second section was snap-frozen in liquid nitrogen for quantitative polymerase chain reaction (qPCR) analysis of tight junction proteins (TJP) including claudin-1, claudin-3, occludin, zonula occludens (ZO-1, ZO-2), junctional adhesion molecules (JAM) and E-cadherin. Additionally genes involved in enterocyte protection including glucagon-like peptide (GLP-2), heat-shock protein (HSP-70), intestinal alkaline phosphatase (IAP), mammalian target of rapamycin (mTOR), toll-like receptors (TLR-4), mucin (MUC-2), cluster differentiation (CD-36) and fatty acid-binding protein (FABP-6) were also analysed. Normally distributed data were analysed using one-way analysis of variance ANOVA. Other data were analysed by non-parametric one-way ANOVA. Villus height and crypt depth were increased (p < .05) only in the ileum after fasting for 4.5 and 9 hr compared with non-fasting group. mRNA expression of claudin-3 was significantly reduced in the ileum of birds fasted for 9 and 19.5 hr, suggesting a role in IP modulation. However, all other TJP genes examined were not statistically different from control. Nevertheless, ileal FABP-6 of all fasted groups was significantly reduced, which could possibly be due to reduced bile acid production during fasting.
Asunto(s)
Pollos/fisiología , Privación de Alimentos , Regulación de la Expresión Génica/fisiología , Mucosa Intestinal/fisiología , Animales , Masculino , Permeabilidad , Reacción en Cadena de la Polimerasa/métodos , Reacción en Cadena de la Polimerasa/veterinaria , Factores de Tiempo , TranscriptomaRESUMEN
Increased intestinal permeability (IP) can lead to compromised health in chickens. As there is limited literature on in vivo biomarkers to assess increased IP in chickens, the objective of this study was to identify a reliable biomarker of IP using DSS ingestion and fasting models. Male Ross chickens (n = 48) were reared until day 14 on the floor pen in an animal care facility, randomized into the following groups: control, DSS and fasting (each with n = 16), and then placed in metabolism cages. DSS was administered in drinking water at 0.75% from days 16 to 21, while controls and fasted groups received water. All birds had free access to feed and water except the birds in the fasting group that were denied feed for 19.5 h on day 20. On day 21, all chickens were given two separate oral gavages comprising fluorescein isothiocyanate dextran (FITC-d, 2.2 mg in 1 ml/bird) at time zero and lactulose, mannitol and rhamnose (LMR) sugars (0.25 g L, 0.05 g M and 0.05 g R in 2 ml/bird) at 60 min. Whole blood was collected from the brachial vein in a syringe 90 min post-LMR sugar gavage. Serum FITC-d and plasma LMR sugar concentrations were measured by spectrophotometry and high-performance ion chromatography respectively. Plasma concentrations of intestinal fatty acid binding protein, diamine oxidase, tight junction protein (TJP), d-lactate and faecal α-antitrypsin inhibitor concentration were also analysed by ELISA. FITC-d increased significantly (p < 0.05) after fasting compared with control. L/M and L/R ratios for fasting and L/M ratio for DSS increased compared with control chickens (p < 0.05). TJP in plasma was significantly increased due to fasting but not DSS treatment, compared with controls. Other tests did not indicate changes in IP (p > 0.05). We concluded that FITC-d and LMR sugar tests can be used in chickens to assess changes in IP.
Asunto(s)
Pollos/sangre , Privación de Alimentos , Mucosa Intestinal/efectos de los fármacos , Animales , Biomarcadores , Sulfato de Dextran , Lactulosa/sangre , Masculino , Manitol/sangre , Permeabilidad , Ramnosa/sangreRESUMEN
Increased intestinal permeability (IP) can lead to compromised health. Limited in vivo IP research has been conducted in chickens. The objectives of the current study were to develop a model of increased IP utilizing lipopolysaccharide (LPS Escherichia coli O55:B5) and to evaluate IP changes using the lactulose, mannitol and rhamnose (LMR) sugar permeability test. In addition, fluorescein isothiocyanate dextran (FITC-d), d-lactate, zonula occludens (ZO-1) and diamine oxidase (DAO) permeability tests were employed. Male Ross chickens were reared until day 14 on the floor in an animal care facility and then transferred to individual cages in three separate experiments. In each of experiments 1 and 2, 36 chicks were randomly allocated to receive either saline (control) or LPS (n=18/group). Lactulose, mannitol and rhamnose sugar concentration in blood was measured at 0, 30, 60, 90, 120 and 180 min in experiment 1, at 60, 90 and 120 min in experiment 2 and at 90 min in experiment 3 (n=16/group). Lipopolysaccharide was injected intraperitoneally at doses of 0.5, 1 and 1 mg/kg BW in experiments 1, 2 and 3, respectively, on days 16, 18 and 20, whereas control received sterile saline. On day 21, only birds in experiments 1 and 2 were fasted for 19.5 h. Chicks were orally gavaged with the LMR sugars (0.25 gL, 0.05 gM, 0.05 gR/bird) followed by blood collection (from the brachial vein) as per time point for each experiment. Only in experiment 3, were birds given an additional oral gavage of FITC-d (2.2 mg/ml per bird) 60 min after the first gavage. Plasma d-lactate, ZO-1 and DAO concentrations were also determined by ELISA in experiment 3 (n=10). Administration of LPS did not affect IP as measured by the LMR sugar test compared with control. This was also confirmed by FITC-d and DAO levels in experiment 3 (P>0.05). The plasma levels of d-lactate were decreased (P<0.05). Plasma levels of ZO-1 were increased in the third experiment only and did not change in the first two experiments. Lipopolysaccharide at doses of 0.5 and 1 mg/kg did not increase IP in this model system. In conclusion, the LMR sugar can be detected in blood 90 min after the oral gavage. Further studies are needed for the applicability of LMR sugars tests.
Asunto(s)
Pollos/fisiología , Escherichia coli/química , Lipopolisacáridos/administración & dosificación , Modelos Biológicos , Amina Oxidasa (conteniendo Cobre)/sangre , Amina Oxidasa (conteniendo Cobre)/metabolismo , Animales , Dextranos/análisis , Dextranos/metabolismo , Fluoresceína-5-Isotiocianato/análogos & derivados , Fluoresceína-5-Isotiocianato/análisis , Fluoresceína-5-Isotiocianato/metabolismo , Intestinos/fisiología , Ácido Láctico/sangre , Ácido Láctico/metabolismo , Lactulosa/sangre , Lactulosa/metabolismo , Masculino , Manitol/sangre , Manitol/metabolismo , Permeabilidad/efectos de los fármacos , Distribución Aleatoria , Ramnosa/sangre , Ramnosa/metabolismo , Uniones Estrechas/metabolismoRESUMEN
A direct process for manufacturing polymer carbon nanotube (CNT)-based composite yarns is reported. The new approach is based on a modified dry spinning method of CNT yarn and gives a high alignment of the CNT bundle structure in yarns. The aligned CNT structure was combined with a polymer resin and, after being stressed through the spinning process, the resin was cured and polymerized, with the CNT structure acting as reinforcement in the composite. Thus the present method obviates the need for special and complex treatments to align and disperse CNTs in a polymer matrix. The new process allows us to produce a polymer/CNT composite with properties that may satisfy various engineering specifications. The structure of the yarn was investigated using scanning electron microscopy coupled with a focused-ion-beam system. The tensile behavior was characterized using a dynamic mechanical analyzer. Fourier transform infrared spectrometry was also used to chemically analyze the presence of polymer on the composites. The process allows development of polymer/CNT-based composites with different mechanical properties suitable for a range of applications by using various resins.
Asunto(s)
Nanocompuestos/química , Nanotecnología/instrumentación , Nanotecnología/métodos , Nanotubos de Carbono/química , Polímeros/química , Módulo de Elasticidad , Elastómeros/química , Microscopía Electrónica de Rastreo , Nanocompuestos/ultraestructura , Nanotubos de Carbono/ultraestructura , Polímeros/síntesis química , Poliuretanos/química , Espectroscopía Infrarroja por Transformada de Fourier , Estrés Mecánico , Propiedades de Superficie , Temperatura , Resistencia a la TracciónRESUMEN
AIMS: To evaluate the efficacy of bovine lactoferrin (BLf), recombinant human lactoferrin (rHLf) and desferrioxamine against Helicobacter pylori in vitro and in mice and also to determine whether BLf or rHLf alter gastric inflammation. METHODS AND RESULTS: In vitro: Broth dilution susceptibility tests were performed using different concentrations of desferrioxamine, BLf and rHLf. Murine trials: In the prevention trial, C57BL/6 female mice were treated with BLf or rHLF, and then infected with the SS1 strain of H. pylori. In the treatment trial, mice were gavaged with either BLf, rHLf or desferrioxamine. In addition, gastric myeloperoxidase activity (MPO) was measured to assess gastric inflammation. Desferoxamine was found to have a direct bactericidal effect, while BLf and rHLf only partially suppressed H. pylori growth in vitro. However, in both prevention and treatment trials all three forms of treatment failed to reduce H. pylori load in mice. Gastric MPO activity and H. pylori load were noted to be higher with lactoferrin treatments. CONCLUSIONS: Our study does not support the use of BLf or rHLF in the treatment of human H. pylori infection. Interestingly, H. pylori growth and gastric inflammation appear to be enhanced by lactoferrin treatment. SIGNIFICANCE AND IMPACT OF THE STUDY: The mouse model is ideal for testing novel H. pylori eradicating agents.
Asunto(s)
Antibacterianos/farmacología , Deferoxamina/farmacología , Mucosa Gástrica/inmunología , Infecciones por Helicobacter/tratamiento farmacológico , Helicobacter pylori/crecimiento & desarrollo , Lactoferrina/farmacología , Animales , Antibacterianos/efectos adversos , Deferoxamina/efectos adversos , Femenino , Mucosa Gástrica/efectos de los fármacos , Mucosa Gástrica/microbiología , Infecciones por Helicobacter/inmunología , Infecciones por Helicobacter/microbiología , Helicobacter pylori/efectos de los fármacos , Humanos , Inflamación/tratamiento farmacológico , Inflamación/microbiología , Lactoferrina/efectos adversos , Ratones , Resultado del TratamientoRESUMEN
Zinc (Zn) and its binding protein metallothionein (MT) have been proposed to suppress the disease activity in ulcerative colitis. To determine the role of Zn and MT in the dextran sulfate sodium (DSS)-induced model of colitis in mice, a DSS dose-response study was conducted in male C57BL/6 wild-type (MT+/+) and MT-null (MT-/-) mice by supplementing 2%, 3%, and 4% DSS in the drinking water for 6 days. In the intervention study, colitis was induced with 2% DSS, Zn (24 mg/ml as ZnO) was gavaged (0.1 ml) daily, concurrent with DSS administration, and the disease activity index (DAI) was scored daily. Histology, MT levels, and myeloperoxidase (MPO) activity were determined. DAI was increased (P<0.05) by 16% and 21% with 3% and 4% concentrations of DSS, respectively, compared to 2%, evident after 5 days of DSS administration. MPO activity was increased in MT+/+ compared to MT-/- mice and those receiving DSS. Zn administration had a 50% (P<0.05) lower DAI compared to DSS alone. Zn partially prevented the distal colon of MT+/+ by 47% from DSS-induced damage compared to MT-/- mice. MT did not prevent DSS-induced colitis and Zn was partially effective in amelioration of DSS-induced colitis.
Asunto(s)
Colitis Ulcerosa/tratamiento farmacológico , Metalotioneína/uso terapéutico , Oligoelementos/uso terapéutico , Zinc/uso terapéutico , Animales , Colitis Ulcerosa/inducido químicamente , Colitis Ulcerosa/metabolismo , Colon/efectos de los fármacos , Colon/metabolismo , Colon/patología , Sulfato de Dextran/administración & dosificación , Sulfato de Dextran/toxicidad , Modelos Animales de Enfermedad , Relación Dosis-Respuesta a Droga , Estudios de Seguimiento , Masculino , Metalotioneína/farmacocinética , Ratones , Ratones Endogámicos C57BL , Peroxidasa/metabolismo , Sustitutos del Plasma/administración & dosificación , Sustitutos del Plasma/toxicidad , Índice de Severidad de la Enfermedad , Espectrofotometría , Oligoelementos/farmacocinética , Resultado del Tratamiento , Zinc/farmacocinéticaRESUMEN
BACKGROUND: Mucosal damage by H. pylori infection is mainly caused by neutrophils producing large quantities of reactive oxygen species (ROS). Metallothionein (MT) an intracellular, low-molecular, cysteine-rich protein, which is inducible by dietary zinc (Zn), has been implicated in sequestering ROS. This study examines the effects of Zn supplementation on Helicobacter colonisation and associated gastritis and the relationship with gastric MT levels. METHODS: C57Bl/6 mice were inoculated with either 10(8) H. pylori or H. felis and were infected for 4 weeks or 6 and 12 weeks, respectively. Mice infected with H. pylori (4 weeks) or H. felis (6 weeks) were treated with either Zn acetate (ZnA; 1 mg/ml), or Zn sulphate (ZnSO4; 5 mg/ml) for 2 weeks with 0.1 ml oro-gastric gavage twice daily. H. pylori load and H. felis colonisation density were determined by culture and microscopy, respectively. MT levels and H. felis-induced gastritis were also determined. RESULTS: Zn treatment showed no significant difference in Helicobacter load and gastric MT, however, ZnSO4 treatment showed a significant (p<0.05) increased in gastric MT in H. felis infected mice. Both Zn-treated groups showed a significant (p<0.05) difference in gastritis score in the antrum of the stomach within the basal and submucosal compartments compared to H. felis-infected controls. CONCLUSIONS: We found that H. felis-induced gastritis can be attenuated by short-term treatment of Zn. This observation suggests that Zn alone may be effective for the suppression of gastric mucosal inflammation induced by Helicobacter.
Asunto(s)
Gastritis/tratamiento farmacológico , Infecciones por Helicobacter/tratamiento farmacológico , Helicobacter felis , Acetato de Zinc/uso terapéutico , Sulfato de Zinc/uso terapéutico , Administración Oral , Animales , Relación Dosis-Respuesta a Droga , Evaluación Preclínica de Medicamentos , Gastritis/etiología , Gastritis/metabolismo , Infecciones por Helicobacter/complicaciones , Infecciones por Helicobacter/metabolismo , Helicobacter felis/aislamiento & purificación , Metalotioneína/metabolismo , Ratones , Ratones Endogámicos C57BL , Estómago/microbiología , Estómago/patología , Acetato de Zinc/administración & dosificación , Sulfato de Zinc/administración & dosificaciónRESUMEN
A new method based on near-infrared (NIR) spectrometry and partial least-squares analysis has been developed for the noninvasive and nondestructive determination of the identity and sequences of amino acid residues in di- and tripeptides. The di- and tripeptides were synthesized from six amino acids with similar structures (Gly, Ala, Leu, Met, Phe, Val) on two different polymer beads (bead with and without a linker) using the solid-phase peptide synthetic method. The developed NIR method is capable of determining the identity of sequences of these di- and tripeptides (with and without the Fmoc protecting group) directly on the polymer beads. It can distinguish not only dipeptides from tripeptides but also peptides with very similar structures (e.g., bead-Gly-Ala-Ala, bead-Gly-Ala-Phe, bead-Gly-Ala-Leu, bead-Gly-Ala-Val, and bead-Gly-Ala-Met). More importantly, the method is capable of distinguishing di- and tripeptides with the same amino acid residues but different sequences (e.g., bead-Gly-Leu-Val from bead-Gly-Val-Leu).
Asunto(s)
Dipéptidos/análisis , Oligopéptidos/análisis , Técnicas Químicas Combinatorias , Dipéptidos/síntesis química , Indicadores y Reactivos , Oligopéptidos/síntesis química , Espectroscopía Infrarroja CortaRESUMEN
A multispectral imaging spectrometer is an instrument that can simultaneously record spectral and spatial information of a sample. Chemical and physical properties of the sample can be elucidated from such images. By synergistic use of an acousto-optic tunable filter and a progressive scan camera capable of snap shot recording it was possible to develop a novel imaging spectrometer with a spatial resolution of a few microns and which can record, grab and store up to 33 images per second (at a function of time) or 16 images per second (as a function of wavelength). This overview article summarizes the instrumentation development of various imaging spectrometers and their applications including its use as the detector for the determination of identity and sequences of peptides synthesized by the combinatorial solid phase method.
Asunto(s)
Oligopéptidos/química , Biblioteca de Péptidos , Secuencia de Aminoácidos , Técnicas Químicas Combinatorias , Diseño de Equipo , Microscopía/instrumentación , Microscopía/métodos , Sensibilidad y Especificidad , Espectrofotometría/instrumentación , Espectrofotometría/métodosRESUMEN
A new multispectral imaging microscope with micrometer spatial resolution and millisecond temporal resolution has been developed. The imaging microscope is based on the use of an acousto-optic tunable filter (AOTF) for spectral tuning and a progressive scan camera capable of snapshot operation for recording. It can operate in two modes: images are recorded as a function of time or wavelength. When operated as a function of time, the microscope is configured so that as many images as possible are recorded, grabbed, and stored per one wavelength. Upon completion, the AOTF is scanned to a new wavelength, and a new set of images are recorded. Up to 33 images/ second (i.e., 30 ms/image) can be recorded in this mode. In the other configuration, the recording wavelength is rapidly scanned (by means of the AOTF) and only one image is rapidly recorded, grabbed, and stored for each wavelength. Because additional time is needed to scan the AOTF, the maximum number of images can be grabbed in this case is 16 frames/s. Preliminary applications of the imaging microscope include measurements of photoinduced changes of a single unit cell in temperature-sensitive cholesteric liquid crystals as a function of time and wavelength. The changes were found to be varied with time and wavelength. Interestingly, the photoinduced changes of unit cells in the liquid crystal are not the same but different from cell to cell. This imaging microscope is particularly useful for measurements of small-size samples that undergo rapid chemical or biochemical reactions, e.g., activities of a single biological cell.
Asunto(s)
Microscopía/métodos , Colesterol/química , Cristalización , Procesamiento de Imagen Asistido por Computador , Microscopía/instrumentaciónRESUMEN
A new method based on the near infrared technique has been developed for the noninvasive and nondestructive determination of the identity and sequences of amino acid residues in small peptides. The method is capable of distinguishing not only peptides with very similar structures (e.g., Gly-Ala-Ala, Gly-Ala-Leu, Leu-Gly-Gly and Gly-Leu-Leu-Gly, Gly-Leu-Gly-Gly, Gly-Gly-Ala-Gly) but also peptides with the same amino acid residues but different sequences (e.g., Gly-Ala-Ala, Ala-Gly-Ala, Ala-Ala-Gly and Gly-Gly-Gly-Ala, Gly-Gly-Ala-Gly).
Asunto(s)
Péptidos/análisis , Espectroscopía Infrarroja Corta/instrumentación , Espectroscopía Infrarroja Corta/métodos , Sensibilidad y EspecificidadRESUMEN
Measurements of two-photon-excited fluorescence (TPF) of fluorescein and Rhodamine 6G in various solvents were performed with a continuous-wave (cw) laser for excitation and an acousto-optic tunable filter for spectral dispersion. Interestingly, the cw laser excitation produced an unwanted thermal-lens effect when the measurements were performed in solvents that absorb the excitation laser light (e.g., alcohols and water, because these solvents absorb the 780-nm excitation light through the overtone and combination transitions of the O-H group). The defocusing effect of the thermal lens leads to a decrease in the TPF signal. Because the strength of the thermal lens depends on the thermo-optical properties (dn/dT and thermal conductivity) of the solvent, its interference makes the effect of solvents on the TPF much different from those on one-photon-excited fluorescence. However, the thermal-lens interference will not limit the application of this cw laser excited TPF technique because, even when measurements were performed in solvents that absorb cw excitation laser light, the thermal-lens interference was observed only in solvents such as nonpolar organic solvents that have relatively better thermo-optical properties. Interference was not observed in water, which is the most widely used solvent for the TPF technique (because water has poor thermo-optical properties).
RESUMEN
BACKGROUND: Zinc (Zn) is protective and enhances epithelial repair in gut diseases. In this study we investigate the localization and distribution of Zn and its binding protein, metallothionein (MT), in the gut of rats fed diets varying in Zn content. METHODS: Male-Sprague Dawley rats were fed low, normal, high, or excess Zn in their diets (10, 100, 400, or 1000 mg Zn/kg, respectively) and killed 7 days later. Blood, liver, and gut tissues were collected. Tissue Zn was determined with atomic absorption spectrophotometery and MT with a Cd/haem affinity assay. Zn and MT were immunohistochemically localized in the small-intestinal wall with zinquin and an anti-MT antibody. RESULTS: Most Zn in the intestinal wall was present in the mucosal scrapings, with 94% membrane-bound and 6% cytosolic, irrespective of dietary Zn. MT levels increased in all gut regions at dietary Zn levels above 100 mg Zn/kg. MT was 40% higher in the ileum than in other gut regions in rats fed low- and normal-Zn diets. The Zn content of the ileum was also 20% higher than that of other gut regions in rats fed low-, normal-, or high-Zn diets. Zn and MT were colocalized in the base of the intestinal crypts, most visibly in the ileum. CONCLUSION: Mucosal cytosolic Zn and MT concentrations are increased only at high or excessive Zn intakes in all gut regions except the ileum, which can respond to a lower Zn intake. As the cytosolic Zn pool most likely influences mucosal protection and repair mechanisms, it is proposed that an increased MT may indicate the adequacy of oral Zn therapy in gut disease.
Asunto(s)
Intestino Delgado/metabolismo , Metalotioneína/análisis , Zinc/análisis , Zinc/farmacocinética , Músculos Abdominales/metabolismo , Análisis de Varianza , Animales , Colorantes Fluorescentes , Inmunohistoquímica , Mucosa Intestinal/metabolismo , Intestino Delgado/patología , Hígado/metabolismo , Masculino , Metalotioneína/inmunología , Microscopía Fluorescente , Quinolonas , Ratas , Ratas Sprague-Dawley , Espectrofotometría Atómica , Compuestos de Tosilo , Zinc/administración & dosificaciónRESUMEN
A near-infrared (NIR) multispectral imaging spectrometer was used to monitor solid-phase peptide synthesis. This imaging spectrometer has fast scanning ability and high sensitivity because it is based on an acousto-optic tunable filter and a NIR InGaAs focal plane array camera. This NIR imaging instrument possesses all the advantages of conventional NIR spectrometers; namely, it can be used for noninvasive monitoring of the reactions and identification of the products during the solid-phase peptide synthesis of glycine, alanine, and valine mediated by aminomethylstyrene resin beads. The reaction was determined by monitoring either the decrease of the band at 1529 nm, which is due to the amine group on the beads, or the increase of the amide band generated at 1483 nm. The amine band at 1529 nm was also used to determine the presence of the Fmoc protecting groups and the efficiency of its removal. More importantly, this NIR imaging spectrometer has additional features that conventional NIR spectrometers cannot offer; namely, its ability to measure spectra at different positions within a sample. This feature was utilized for the first demonstration in which reactions of three different solid-phase peptide syntheses (in a three-compartment cell) were simultaneously monitored. As expected, the kinetics obtained for three reactions are similar to those obtained when the each of the reactions was individually determined. In this study, data recorded by 16 x 16 pixels were used to calculate a spectrum for each sample. However, a relatively good spectrum can be obtained by using data recorded by a single pixel. Since the NIR camera used in this camera is equipped with 240 x 320 pixels, this NIR mutispectral imaging technique is not limited to the three-compartment cell used in this study but rather can be used as the detection method for the solid-phase peptide synthesis in combinatorial chemistry.
Asunto(s)
Péptidos/síntesis química , Química Orgánica/métodos , Espectroscopía Infrarroja CortaRESUMEN
The kinetics of curing of an epoxy resin by amine was studied using a near-infrared (NIR) multispectral imaging spectrometer. This imaging spectrometer is capable of sensitively and rapidly recording NIR spectral images of a sample because it was constructed with an acousto-optic tunable filter and an InGaAs focal plane array NIR camera. The high sensitivity and fast scanning ability of the spectrometer make it suitable for kinetic determination of fast reactions. Additionally, it has features that conventional NIR spectrometers cannot offer, namely, its ability to provide kinetic information at different positions within a sample. Furthermore, the high spatial resolution and sensitivity of the InGaAs camera make it possible to determine the kinetics from data collected by a single pixel in the camera. The kinetics of curing of epoxy by amine, determined by this multispectral imaging instrument, show that the reaction rates within the sample are very inhomogeneous. Because of this kinetic inhomogeneity, differences in the degrees of cure at different positions within the sample can be as high as 37% when data from only a single pixel were used for calculation. The inhomogeneity was not be observed if an average of a large number of pixels were used.
Asunto(s)
Compuestos Epoxi/química , Cinética , Espectroscopía Infrarroja CortaRESUMEN
Gut Zn homeostatic responses to low, replete, and excess dietary Zn (10, 150, and 400 mg Zn/kg, respectively) were compared in mice with (MT+/+) and without (MT-/-) metallothionein (MT) expression. MT concentrations decreased progressively from stomach (12.9 nmol Cd bound/g) to colon (4.6 nmol Cd bound/g). Small intestinal MT was increased in mice fed the 400-mg Zn/kg diet (+130%, duodenum; +56%, jejunum; +29%, terminal ileum), but not in the stomach, cecum and colon. Zn concentrations were much higher in the distal gut at increasing Zn intakes in MT+/+ mice but to a lesser extent in MT-/- mice. On the 10-mg Zn/kg diet, MT-/- mice had 45% more Zn in the jejunum/ileum than MT+/+ mice. In fasted (20 h) mice, Zn concentrations in all gut regions were similar to those of MT+/+ mice fed the 10-mg Zn/kg diet, irrespective of prior Zn intake or genotype. Liver MT quadrupled in mice fasted after the 10-mg Zn/kg diet but only doubled after the 400-mg Zn/kg diet, a trend also present in gut MT. Glucagon administration stimulated gut as well as liver MT, implicating it as a major component of the MT response to fasting. MT-/- mice had five times more variation than MT+/+ mice in plasma Zn over all dietary groups. Together, these findings demonstrate that without MT, there is little modification of regional gut Zn concentrations in response to extremes of dietary Zn and poorer regulation of Zn homeostasis.
Asunto(s)
Sistema Digestivo/metabolismo , Metalotioneína/metabolismo , Zinc/metabolismo , Animales , Dieta , Glucagón/administración & dosificación , Metalotioneína/genética , Ratones , Ratones Endogámicos C57BL , Ratones Noqueados , Distribución Tisular , Zinc/administración & dosificación , Zinc/sangreRESUMEN
A new multispectral imaging spectrometer capable of simultaneously recording spectral images in the visible and near-infrared has been developed. In this instrument, an acoustooptic tunable filter is used to diffract an unpolarized incident light into two diffracted beams with orthogonal polarization; one of them is detected by a silicon camera for the visible region while the other beam is detected in the near-infrared region (from 1 to 1.7 microns) with a NIR camera. The imaging spectrometer is sensitive, inexpensive, and field deployable because it is based on the recently available InGaAs focal plane arrays camera, which is low cost and can be sensitively operated at room temperature. Preliminary applications of the imaging spectrometer include measurements of the visible and NIR absorption spectra of ink used to print U.S. currency. Such results may help to characterize samples as well as to control and to ensure the quality of the samples during the production processes. More important are the results obtained on ethylene/vinyl acetate copolymers. The NIR spectral images obtained clearly indicate that these copolymers exhibit a high degree of chemical inhomogeneity. Because of the possibility of inhomogeneity, it is very important that the homogeneity of polymers or copolymers be thoroughly understood before the NIR methods, especially those based on NIR spectrometers equipped with a single-element detector, are used for measurements.
Asunto(s)
Polímeros/análisis , Diagnóstico por Imagen , Espectroscopía Infrarroja CortaRESUMEN
A novel integrated acoustooptic tunable filter (IAOTF) has been developed. This tunable filter is based on the Bragg interactions between waveguide and surface acoustic waves. Compared to (bulk) AOTF, its advantage include all-fiber construction, smaller size, narrower spectral resolution (1.7 nm), higher diffraction efficiency (37%), and lower rf power requirement (150 mW). A relatively narrow spectral tuning range (about 80 nm) is the only drawback for this integrated tunable filter. However, this disadvantage was overcome by judiciously using the filter for measurements in which its tuning range is coincident with the light source and also with absorption bands of analytes. In fact, an all-fiber, compact, high-throughput near-infrared spectrophotometer has been successfully constructed by synergistic use of this integrated AOTF and the erbium-doped fiber amplifier (EDFA), which has been shown to provide high intensity and wide spectral band-width in the near-infrared region from 1500 to 1600 nm. This spectral region is particularly useful for the determination of samples which have O-H and/or N-H groups. The all-fiber nature, compactness, high throughput, and high sensitivity of this spectrophotometer make it particularly suitable for on-line and real-time detection of trace gases in hostile environments, including leak detection of monomethylhydrazine (at a limit of detection of 191 ppm), which is often used as the hypergolic propellant for the space shuttle thruster systems.