Discrimination between washed Arabica, natural Arabica and Robusta coffees by using near infrared spectroscopy, electronic nose and electronic tongue analysis.

BACKGROUND: The aim of this study is to investigate the feasibility of a 'holistic' approach, using near infrared (NIR) spectroscopy and electronic devices (electronic nose and electronic tongue), as instrumental tools for the classification of different coffee varieties. Analyses were performed on green coffee, on ground roasted coffee and on coffee beverage. Principal component analysis was applied on spectral and sensory data to uncover correlations between samples and variables. After variable selection, linear discriminant analysis was used to classify the samples on the basis of the three coffee classes: Robusta, natural Arabica and washed Arabica. RESULTS: Linear discriminant analysis demonstrates the practicability of this approach: the external test set validation performed with NIR data showed 100% of correctly classified samples. Moreover, a satisfying percentage of correct classification in cross-validation was obtained for the electronic devices: the average values of correctly classified samples were 81.83% and 78.76% for electronic nose and electronic tongue, respectively. CONCLUSION: NIR spectroscopy was shown to be a very reliable and useful tool to classify coffee samples in a fast, clean and inexpensive way compared to classical analysis, while the electronic devices could assume the role of investigating techniques to depict the aroma and taste of coffee samples.

Propranolol, a ß-adrenoceptor antagonist, worsens liver injury in a model of non-alcoholic steatohepatitis.

Prazosin an α1-adrenoceptor (AR) antagonist has been shown to reduce liver injury in a mouse model of non-alcoholic steatohepatitis (NASH) and is suggested as a potential treatment of NASH especially given its concomitant anti-fibrotic properties. The effect however, of ß-AR blockade in non-cirrhotic NASH is unknown and is as such investigated here. In the presence of the ß-blocker propranolol (PRL), mice fed normal chow or a half methionine and choline deficient diet, supplemented with ethionine (HMCDE), to induce NASH, showed significantly enhanced liver injury, as evidenced by higher hepatic necrosis scores and elevated serum aminotransferases (ALT). Mechanistically, we showed that murine hepatocytes express α and ß adrenoceptors; that PRL directly induces hepatocyte injury and death as evidenced by increased release of lactate dehydrogenase, FASL and TNF-α from hepatocytes in the presence of PRL; and that PRL activated the apoptotic pathway in primary hepatocyte cultures, as indicated by upregulation of Fas receptor and caspase-8 proteins. The ß-AR antagonist PRL therefore appears to enhance liver injury through induction of hepatocyte death via the death pathway. Further studies are now required to extrapolate these findings to humans but meanwhile, ß-AR antagonists should be avoided or used with caution in patients with non-cirrhotic NASH as they may worsen liver injury.

Studies on proofing of yeasted bread dough using near- and mid-infrared spectroscopy.

Dough proofing is the resting period after mixing during which fermentation commences. Optimum dough proofing is important for production of high quality bread. Near- and mid-infrared spectroscopies have been used with some success to investigate macromolecular changes during dough mixing. In this work, both techniques were applied to a preliminary study of flour doughs during proofing. Spectra were collected contemporaneously by NIR (750-1100 nm) and MIR (4000-600 cm(-1)) instruments using a fiberoptic surface interactance probe and horizontal ATR cell, respectively. Studies were performed on flours of differing baking quality; these included strong baker's flour, retail flour, and gluten-free flour. Following principal component analysis, changes in the recorded spectral signals could be followed over time. It is apparent from the results that both vibrational spectroscopic techniques can identify changes in flour doughs during proofing and that it is possible to suggest which macromolecular species are involved.

Evaluation of freshness decay of minced beef stored in high-oxygen modified atmosphere packaged at different temperatures using NIR and MIR spectroscopy.

Meat freshness has been monitored by various microbiological, chemical and sensorial indices. However, these methods are slow and not suited to automation. Infrared spectroscopy is one of the most convenient analytical tools which could be used to monitor the evolution of food quality. The aim of this work was to investigate the ability of both NIR (Near Infrared) and MIR (Mid Infrared) spectroscopy to follow meat freshness decay. The minced beef was packaged in high-oxygen modified atmosphere (30% CO2 and 70% O2) and stored at three temperatures. Spectra were collected by Fourier-Transformation (FT)-NIR and FT-IR instruments. PCA, applied to the data, was able to discriminate samples on the basis of storage time and temperature. The modelling of PC scores versus time allowed the setting of the time of initial freshness decay for the samples (6-7 days at 4.3°C, 2-3 days at 8.1°C and less than 1 day at 15.5°C).

Chemometrical strategies for feature selection and data compression applied to NIR and MIR spectra of extra virgin olive oils for cultivar identification.

The possibility provided by Chemometrics to extract and combine (fusion) information contained in NIR and MIR spectra in order to discriminate monovarietal extra virgin olive oils according to olive cultivar (Casaliva, Leccino, Frantoio) has been investigated. Linear discriminant analysis (LDA) was applied as a classification technique on these multivariate and non-specific spectral data both separately and jointly (NIR and MIR data together). In order to ensure a more appropriate ratio between the number of objects (samples) and number of variables (absorbance at different wavenumbers), LDA was preceded either by feature selection or variable compression. For feature selection, the SELECT algorithm was used while a wavelet transform was applied for data compression. Correct classification rates obtained by cross-validation varied between 60% and 90% depending on the followed procedure. Most accurate results were obtained using the fused NIR and MIR data, with either feature selection or data compression. Chemometrical strategies applied to fused NIR and MIR spectra represent an effective method for classification of extra virgin olive oils on the basis of the olive cultivar.

Preliminary study on application of mid infrared spectroscopy for the evaluation of the virgin olive oil "freshness".

The freshness of virgin olive oils (VOO) from typical cultivars of Garda regions was evaluated by attenuated total reflectance (ATR) and Fourier transform infrared (FTIR) spectroscopy, in combination with multivariate analysis. The olive oil freshness decreased during storage mainly because of oxidation processes. In this research, 91 virgin olive oils were packaged in glass bottles and stored either in the light or in the dark at room temperature for different periods. The oils were analysed, before and after storage, using both chemical methods and spectroscopic technique. Classification strategies investigated were partial least square discriminant analysis (PLS-DA), linear discriminant analysis (LDA), and soft independent modelling of class analogy (SIMCA). The results show that ATR-MIR spectroscopy is an interesting technique compared with traditional chemical index in classifying olive oil samples stored in different conditions. In fact, the FTIR PCA results allowed a better discrimination among fresh and oxidized oils, than samples separation obtained by PCA applied to chemical data. Moreover, the results obtained by the different classification techniques (PLS-DA, LDA, SIMCA) evidenced the ability of FTIR spectra to evaluate the olive oil freshness. FTIR spectroscopy results are in agreement with classical methods. The spectroscopic technique could be applied for the prediction of VOOs freshness giving information related to chemical modifications. The great advantages of this technique, compared to chemical analysis, are related to rapidity, non-destructive characteristics and low cost per sample. In conclusion, ATR-MIR represents a reliable, cheap and fast classification tool able to assess the freshness of virgin olive oils.

Human hepatic progenitor cells express vasoactive intestinal peptide receptor type 2 and receive nerve endings.

BACKGROUND: We recently showed that human hepatic progenitor cells (HPCs) express muscarinic acetylcholine (Ach) receptor subtype 3 and that--following liver transplantation--HPC numbers are significantly reduced. To further elaborate on this, we examined whether HPC also express receptors for vasoactive intestinal peptide (VIP), which, besides Ach, also is an important parasympathetic neurotransmitter. VIP expressing nerves are known to be present in the liver. METHODS: We performed immunohistochemistry for VIP receptor subtypes 1 and 2 (VIPR1 and 2), on sections of normal and diseased human liver (n=17), and double staining for VIPR2 and known HPC markers. We performed RT-PCR for VIPR1 and 2 on total RNA from purified rat HPC. To document the probability of direct interaction, we also performed double immunostaining for nerve markers and HPC markers on human liver sections. RESULTS: VIPR2 immunostaining was clearly positive in HPC and reactive bile ductules on paraffin-embedded and frozen tissue sections. We could not demonstrate VIPR1 protein expression in the liver, with either of two VIPR1 antibodies tested. The presence of VIPR2 mRNA in HPC was confirmed by RT-PCR. Nerve endings were shown to abut on reactive bile ductules. CONCLUSION: We show here for the first time that HPC express VIPR2 and receive nerve endings. These features, and the fact that HPC numbers are influenced by the presence or absence of the autonomic innervation of the liver, suggest a direct interaction.

The vagal nerve stimulates activation of the hepatic progenitor cell compartment via muscarinic acetylcholine receptor type 3.

In the rat the hepatic branch of the nervus vagus stimulates proliferation of hepatocytes after partial hepatectomy and growth of bile duct epithelial cells after bile duct ligation. We studied the effect of hepatic vagotomy on the activation of the hepatic progenitor cell compartment in human and rat liver. The number of hepatic progenitor cells and atypical reactive ductular cells in transplanted (denervated) human livers with hepatitis was significantly lower than in innervated matched control livers and the number of oval cells in vagotomized rat livers with galactosamine hepatitis was significantly lower than in livers of sham-operated rats with galactosamine hepatitis. The expression of muscarinic acetylcholine receptors (M1-M5 receptor) was studied by immunohistochemistry and reverse transcriptase-polymerase chain reaction. In human liver, immunoreactivity for M3 receptor was observed in hepatic progenitor cells, atypical reactive ductules, intermediate hepatocyte-like cells, and bile duct epithelial cells. mRNA for the M1-M3 and the M5 receptor, but not the M4 receptor, was detected in human liver homogenates. In conclusion, the hepatic vagus branch stimulates activation of the hepatic progenitor cell compartment in diseased liver, most likely through binding of acetylcholine to the M3 receptor expressed on these cells. These findings may be of clinical importance for patients with a transplant liver.

Hepatic immunohistochemical staining with a monoclonal antibody against HCV-E2 to evaluate antiviral therapy and reinfection of liver grafts in hepatitis C viral infection.

BACKGROUND/AIMS: A simple and reproducible hepatic immunohistochemical staining (IHS) for hepatitis C virus (HCV) is not available. We aimed to validate hepatic IHS with monoclonal antibody (Mab) IG222, directed against the HCV-envelope 2 (E2) protein. METHODS: A three-step indirect immunoperoxidase method was used for frozen sections and a two-step indirect EnVision technique was used for paraffin-embedded sections. RESULTS: Naturally or in vitro HCV infected primary human hepatocytes were immunoreactive to HCV-E2. In the patient study (n=253), IHS had a sensitivity of 96% and a specificity of 91%. Six patients who showed positivity in the liver with Mab IG222, but remained anti-HCV and HCV-RNA negative, had hepatitis C-like changes in their liver biopsy. In one patient HCV-RNA could be detected in the liver biopsy. We confirmed early graft reinfection in patients transplanted for HCV-related disease (34 patients with serial biopsies). Treatment for acute cellular rejection with steroids was associated with an increase in staining intensity. In nine patients with clearance of HCV-RNA during antiviral therapy, seven achieved negativation of immunoreactivity and two a marked reduction. CONCLUSIONS: The IHS with Mab IG222 is an accurate tool for diagnosis and clinical management of chronic hepatitis C.

Sympathetic nervous system inhibition increases hepatic progenitors and reduces liver injury.

Recovery from liver damage might be enhanced by encouraging repopulation of the liver by endogenous hepatic progenitor cells. Oval cells are resident hepatic stem cells that promote liver regeneration and repair. Little is known about the mediators that regulate the accumulation of these cells in the liver. Parasympathetic nervous system inhibition reduces the number of oval cells in injured livers. The effect of sympathetic nervous system (SNS) inhibition on oval cell number is not known. Adrenergic inhibition mobilizes hematopoietic precursors into the circulation and has also been shown to promote liver regeneration. Thus, we hypothesized that SNS inhibition would promote hepatic accumulation of oval cells and reduce liver damage in mice fed antioxidant-depleted diets to induce liver injury. Our results confirm this hypothesis. Compared with control mice that were fed only the antioxidant-depleted diets, mice fed the same diets with prazosin (PRZ, an alpha-1 adrenoceptor antagonist) or 6-hydroxydopamine (6-OHDA, an agent that induces chemical sympathectomy) had significantly increased numbers of oval cells. Increased oval cell accumulation was accompanied by less hepatic necrosis and steatosis, lower serum aminotransferases, and greater liver and whole body weights. Neither PRZ nor 6-OHDA affected the expression of cytokines, growth factors, or growth factor receptors that are known to regulate progenitor cells. In conclusion, stress-related sympathetic activity modulates progenitor cell accumulation in damaged livers and SNS blockade with alpha-adrenoceptor antagonists enhances hepatic progenitor cell accumulation.

Norepinephrine induces hepatic fibrogenesis in leptin deficient ob/ob mice.

Leptin's actions on certain cells require a leptin-inducible neurotransmitter, norepinephrine (NE). NE modulates hepatic fibrosis. Therefore, decreased NE may explain why leptin deficiency inhibits hepatic fibrosis. We manipulated adrenergic activity in leptin-deficient ob/ob mice, leptin-sufficient, dopamine beta-hydroxylase deficient (Dbh(-/-)) mice, and HSC cultures to determine if leptin requires NE to activate HSC and induce hepatic fibrosis. ob/ob mice have chronic liver injury, but reduced numbers of HSC. Supplemental leptin increases HSC, suggesting that leptin-dependent, injury-related factors permit expansion of HSC populations. NE also increases HSC numbers and activation, normalizing fibrogenesis. When fed hepatotoxic diets, NE-deficient Dbh(-/-) mice fail to accumulate activated HSC and have impaired fibrogenesis unless treated with adrenergic agonists. NE acts directly on HSC to modulate leptin's actions because leptin increases HSC proliferation and prazosin, an alpha-adrenoceptor antagonist, inhibits this. Thus, leptin permits injury-related increases in adrenergic activity and requires NE to activate HSC and induce hepatic fibrogenesis.