The ß-galactosidase immobilization protocol determines its performance as catalysts in the kinetically controlled synthesis of lactulose.

In this paper, 3 different biocatalysts of ß-galactosidase from Kluyveromyces lactis have been prepared by immobilization in chitosan activated with glutaraldehyde (Chi_Glu_Gal), glyoxyl agarose (Aga_Gly_Gal) and agarose coated with polyethylenimine (Aga_PEI_Gal). These biocatalysts have been used to catalyze the synthesis of lactulose from lactose and fructose. Aga-PEI-Gal only produces lactulose at 50 °C, and not at 25 or 37 °C, Aga_Gly_Gal was unable to produce lactulose at any of the assayed temperatures while Chi_Glu_Gal produced lactulose at all assayed temperatures, although a lower yield was obtained at 25 or 37 °C. The pre-incubation of this biocatalyst at 50 °C permitted to obtain similar yields at 25 or 37 °C than at 50 °C. The use of milk whey instead of pure lactose and fructose produced an improvement in the yields using Aga_PEI_Gal and a decrease using Chi_Glu_Gal. The operational stability also depends on the reaction medium and of biocatalyst. This study reveals how enzyme immobilization may greatly alter the performance of ß-galactosidase in a kinetically controlled manner, and how medium composition influences this performance due to the kinetic properties of ß-galactosidase.

Imaging and immunometabolic phenotyping uncover changes in the hepatic immune response in the early phases of NAFLD.

BACKGROUND & AIMS: The precise determination of non-alcoholic fatty liver disease (NAFLD) onset is challenging. Thus, the initial hepatic responses to fat accumulation, which may be fundamental to our understanding of NAFLD evolution and clinical outcomes, are largely unknown. Herein, we chronologically mapped the immunologic and metabolic changes in the liver during the early stages of fatty liver disease in mice and compared this with human NAFLD samples. METHODS: Liver biopsies from patients with NAFLD (NAFLD activity score [NAS] 2-3) were collected for gene expression profiling. Mice received a high-fat diet for short periods to mimic initial steatosis and the hepatic immune response was investigated using a combination of confocal intravital imaging, gene expression, cell isolation, flow cytometry and bone marrow transplantation assays. RESULTS: We observed major immunologic changes in patients with NAS 2-3 and in mice in the initial stages of NAFLD. In mice, these changes significantly increased mortality rates upon drug-induced liver injury, as well as predisposing mice to bacterial infections. Moreover, deletion of Toll-like receptor 4 in liver cells dampened tolerogenesis, particularly in Kupffer cells, in the initial stages of dietary insult. CONCLUSION: The hepatic immune system acts as a sentinel for early and minor changes in hepatic lipid content, mounting a biphasic response upon dietary insult. Priming of liver immune cells by gut-derived Toll-like receptor 4 ligands plays an important role in liver tolerance in initial phases, but continuous exposure to insults may lead to damage and reduced ability to control infections. LAY SUMMARY: Fatty liver is a very common form of hepatic disease, leading to millions of cases of cirrhosis every year. Patients are often asymptomatic until becoming very sick. Therefore, it is important that we expand our knowledge of the early stages of disease pathogenesis, to enable early diagnosis. Herein, we show that even in the early stages of fatty liver disease, there are significant alterations in genes involved in the inflammatory response, suggesting that the hepatic immune system is disturbed even following minor and undetectable changes in liver fat content. This could have implications for the diagnosis and clinical management of fatty liver disease.

Allogeneic Mesenchymal Stem Cell Transplantation in Dogs With Keratoconjunctivitis Sicca.

Keratoconjunctivitis sicca (KCS) is a dysfunction in tear production associated with clinical signs, which include conjunctival hyperemia, ocular discharge, discomfort, pain, and, eventually, corneal vascularization and pigmentation. Immunosuppressive drugs are routinely administrated for long periods to treat KCS but with side effects and limited results. Evaluation of the clinical benefits of intralacrimal transplantation of allogeneic mesenchymal stem cells (MSCs) in dogs with mild-moderate and severe KCS was done. A total of 24 eyes with KCS from 15 dogs of different breeds were enrolled in the present study. A single transplantation of MSCs (1 × 106) directly into lacrimal glands (dorsal and third eyelid) was performed. The Schirmer tear tests (STTs) and ocular surface improvements were used to assess short- and long-term effects of these cells. The STTs were carried out on day 0 (before MSCs transplantation) and on days 7, 14, 21, and 28, as well as 6 and 12 months after MSC transplantation. Our data demonstrate that allogeneic MSC transplantation in KCS dogs is safe since no adverse effects were observed immediately after transplantation and in short- and long-term follow-ups. A statistically significant increase in the STT and ocular surface improvements was found in all eyes studied. In all the eyes with mild-moderate KCS, STT values reverted to those of healthy eyes, while in eyes with severe KCS, although complete reversion was not found, there was improvement in tear production and in other clinical signs. Our study shows that a single dose of a low number of MSCs can be used to treat KCS in dogs. In contrast to immunosuppressive drug use, MSC transplantation has an effect over a long period (up to 12 months), even after a single administration, and does not require daily drug administration.

Transposição do ducto parotídeo em cão: relato de caso / Parotid duct transposition in dog: case report / Transpocisión del ducto parotídeo en un perro: relato de caso

La transposición del ducto parotídeo es un procedimiento quirúrgico utilizado como alternativa al tratamiento de la queratoconjuntivitis sicca que es refractaria al tratamiento clínico. Esta abordaje quirúrgica es realizada por medio de la transposición del ducto parotídeo de la cavidad oral para el fórnix conjuntival inferior, permitiendo la sustitución de la lágrima por la saliva. Un canino de 2 años de edad, sin raza definida, porte grande, fue atendido por el servicio de oftalmología del Laboratorio de Oftalmología comparada del Hospital Veterinario da Universidade Federal do Paraná y diagnosticado con queratoconjuntivitis sicca severa probablemente de causa neurogénica. Como el paciente demostró ser refractario al tratamiento clínico instituído previamente, se optó por el procedimiento quirúrgico mencionado anteriormente, utilizando el abordaje transcutáneo (abierto). A pesar de las consecuencias, que incluyeron epífora, blefaritis y deposición de minerales en la córnea, el procedimiento fue bien sucedido, permitiendo la lubricación de la superficie ocular.(AU)

Parotid duct transposition is an alternative surgical approach for the treatment of keratoconjunctivitis sicca when the clinical management has proven unsuccessful. This procedure consists in the transposition of parotid duct from the mouth to the inferior conjunctival fornix to provide lubrification of the eye. A large 2-yearold dog, presented at ophthalmology service of Comparative Veterinary Ophthalmology Laboratory at the Veterinary Hospital of the Federal University of Paraná with severe keratoconjunctivitis sicca with probably a neurogenic cause and refractory to previous medical treatments. There by, the patient was treated surgically by the afore mentioned technique. Although some of the expected complications appeared, including epiphora, blepharitis and mineral deposition in the cornea, the surgery was successful providing lubrication of the eye.(AU)

A transposição do ducto parotídeo é um procedimento cirúrgico utilizado como alternativa para o tratamento da ceratoconjuntivite seca refratária ao tratamento clínico. Esta abordagem cirúrgica é realizada por meio da transposição do ducto parotídeo da cavidade oral para o fórnice conjuntival inferior, permitindo a substituição da lágrima pela saliva. Um cão de 2 anos de idade, sem raça definida, porte grande, foi atendido pelo serviço de oftalmologia do Laboratório de Oftalmologia Comparada do Hospital Veterinário da Universidade Federal do Paraná e diagnosticado com ceratoconjuntivite seca severa de provável causa neurogênica. Como o paciente demonstrou ser refratário ao tratamento clínico previamente instituído, optou-se pelo procedimento cirúrgico mencionado, utilizando a abordagem transcutânea (aberta). Apesar das consequências, que incluíram a epífora, blefarite e deposição de minerais na córnea, o procedimento foi bem-sucedido, permitindo a lubrificação da superfície ocular.(AU)

Transposição do ducto parotídeo em cão: relato de caso / Parotid duct transposition in dog: case report / Transpocisión del ducto parotídeo en un perro: relato de caso

La transposición del ducto parotídeo es un procedimiento quirúrgico utilizado como alternativa al tratamiento de la queratoconjuntivitis sicca que es refractaria al tratamiento clínico. Esta abordaje quirúrgica es realizada por medio de la transposición del ducto parotídeo de la cavidad oral para el fórnix conjuntival inferior, permitiendo la sustitución de la lágrima por la saliva. Un canino de 2 años de edad, sin raza definida, porte grande, fue atendido por el servicio de oftalmología del Laboratorio de Oftalmología comparada del Hospital Veterinario da Universidade Federal do Paraná y diagnosticado con queratoconjuntivitis sicca severa probablemente de causa neurogénica. Como el paciente demostró ser refractario al tratamiento clínico instituído previamente, se optó por el procedimiento quirúrgico mencionado anteriormente, utilizando el abordaje transcutáneo (abierto). A pesar de las consecuencias, que incluyeron epífora, blefaritis y deposición de minerales en la córnea, el procedimiento fue bien sucedido, permitiendo la lubricación de la superficie ocular.

Parotid duct transposition is an alternative surgical approach for the treatment of keratoconjunctivitis sicca when the clinical management has proven unsuccessful. This procedure consists in the transposition of parotid duct from the mouth to the inferior conjunctival fornix to provide lubrification of the eye. A large 2-yearold dog, presented at ophthalmology service of Comparative Veterinary Ophthalmology Laboratory at the Veterinary Hospital of the Federal University of Paraná with severe keratoconjunctivitis sicca with probably a neurogenic cause and refractory to previous medical treatments. There by, the patient was treated surgically by the afore mentioned technique. Although some of the expected complications appeared, including epiphora, blepharitis and mineral deposition in the cornea, the surgery was successful providing lubrication of the eye.

A transposição do ducto parotídeo é um procedimento cirúrgico utilizado como alternativa para o tratamento da ceratoconjuntivite seca refratária ao tratamento clínico. Esta abordagem cirúrgica é realizada por meio da transposição do ducto parotídeo da cavidade oral para o fórnice conjuntival inferior, permitindo a substituição da lágrima pela saliva. Um cão de 2 anos de idade, sem raça definida, porte grande, foi atendido pelo serviço de oftalmologia do Laboratório de Oftalmologia Comparada do Hospital Veterinário da Universidade Federal do Paraná e diagnosticado com ceratoconjuntivite seca severa de provável causa neurogênica. Como o paciente demonstrou ser refratário ao tratamento clínico previamente instituído, optou-se pelo procedimento cirúrgico mencionado, utilizando a abordagem transcutânea (aberta). Apesar das consequências, que incluíram a epífora, blefarite e deposição de minerais na córnea, o procedimento foi bem-sucedido, permitindo a lubrificação da superfície ocular.

Octopamine and Dopamine differentially modulate the nicotine-induced calcium response in Drosophila Mushroom Body Kenyon Cells.

In Drosophila associative olfactory learning, an odor, the conditioned stimulus (CS), is paired to an unconditioned stimulus (US). The CS and US information arrive at the Mushroom Bodies (MB), a Drosophila brain region that processes the information to generate new memories. It has been shown that olfactory information is conveyed through cholinergic inputs that activate nicotinic acetylcholine receptors (nAChRs) in the MB, while the US is coded by biogenic amine (BA) systems that innervate the MB. In this regard, the MB acts as a coincidence detector. A better understanding of the properties of the responses gated by nicotinic and BA receptors is required to get insights on the cellular and molecular mechanisms responsible for memory formation. In recent years, information has become available on the properties of the responses induced by nAChR activation in Kenyon Cells (KCs), the main neuronal MB population. However, very little information exists on the responses induced by aminergic systems in fly MB. Here we have evaluated some of the properties of the calcium responses gated by Dopamine (DA) and Octopamine (Oct) in identified KCs in culture. We report that exposure to BAs induces a fast but rather modest increase in intracellular calcium levels in cultured KCs. The responses to Oct and DA are fully blocked by a VGCC blocker, while they are differentially modulated by cAMP. Moreover, co-application of BAs and nicotine has different effects on intracellular calcium levels: while DA and nicotine effects are additive, Oct and nicotine induce a synergistic increase in calcium levels. These results suggest that a differential modulation of nicotine-induced calcium increase by DA and Oct could contribute to the events leading to learning and memory in flies.