Effects of sphingolipid extracts on the morphological structure and lipid profile in an in vitro model of canine skin

Ceramides (CER) are essential sphingolipids of the stratum corneum (SC) that play an important role in maintaining cutaneous barrier function. Skin barrier defects occur in both human beings and dogs affected with atopic dermatitis, and have been associated with decreased CER concentrations and morphological alterations in the SC. The aim of the present study was to investigate the changes induced by three different sphingolipid extracts (SPE-1, SPE-2 and SPE-3) on the morphological structure and lipid composition of canine skin, using an in vitro model, whereby keratinocytes were seeded onto fibroblast-embedded collagen type I matrix at the air-liquid interface. Cell cultures were supplemented with SPE-1, SPE-2, SPE-3 or vehicle (control) for 14 days. The relative concentrations of lipids were determined by ultra-performance liquid chromatography coupled to mass spectrometry. The ultrastructural morphology of samples was examined by transmission electron microscopy. SPE-1 induced significant elevation in total CERs, CER[NS], CER[NDS], CER[NP], CER[AS], CER[AP], CER[EOS] and CER[EOP] subclasses, whereas SPE-2 induced a significant elevation in total CER, CER[AP] and CER[EOS] compared with control conditions. Ultrastructural analysis revealed an increase in lamellar-lipid structures in the SC of SPE-1-treated samples. The findings demonstrated that SPE-1 stimulates production of CERs, as shown by changes in lipid composition and ultrastructural morphology. Thus, SPE-1 contributes to the formation of a well-organised SC and represents a potential therapeutic target for improving skin barrier function in atopic dermatitis

Glucose and fructose have sugar-specific effects in both liver and skeletal muscle in vivo: a role for liver fructokinase.

We examined glucose and fructose effects on serine phosphorylation levels of a range of proteins in rat liver and muscle cells. For this, healthy adult rats were subjected to either oral glucose or fructose loads. A mini-array system was utilized to determine serine phosphorylation levels of liver and skeletal muscle proteins. A glucose oral load of 125 mg/100 g body weight (G 1/2) did not induce changes in phosphorylated serines of the proteins studied. Loading with 250 mg/100 g body weight of fructose (Fr), which induced similar glycemia levels as G 1/2, significantly increased serine phosphorylation of liver cyclin D3, PI3 kinase/p85, ERK-2, PTP2 and clusterin. The G 1/2 increased serine levels of the skeletal muscle proteins cyclin H, Cdk2, IRAK, total PKC, PTP1B, c-Raf 1, Ras and the β-subunit of the insulin receptor. The Fr induced a significant increase only in muscle serine phosphorylation of PI3 kinase/p85. The incubation of isolated rat hepatocytes with 10 mM glucose for 5 min significantly increased serine phosphorylation of 31 proteins. In contrast, incubation with 10 mM fructose produced less intense effects. Incubation with 10 mM glucose plus 75 µM fructose counteracted the effects of the incubation with glucose alone, except those on Raf-1 and Ras. Less marked effects were detected in cultured muscle cells incubated with 10 mM glucose or 10 mM glucose plus 75 µM fructose. Our results suggest that glucose and fructose act as specific functional modulators through a general mechanism that involves liver-generated signals, like micromolar fructosemia, which would inform peripheral tissues of the presence of either glucose- or fructose-derived metabolites

J-PLUS: A wide-field multi-band study of the M15 globular cluster. Evidence of multiple stellar populations in the RGB

The Javalambre Photometric Local Universe Survey (J-PLUS) provides wide field-of-view images in 12 narrow, intermediate and broad-band filters optimized for stellar photometry. Here we have applied J-PLUS data for the first time for the study of Galactic GCs using science verification data obtained for the very metal-poor GC M\,15. Our J-PLUS data provide low-resolution spectral energy distributions covering the near-UV to the near-IR, allowing us to search for MPs based on pseudo-spectral fitting diagnostics. J-PLUS CMDs are found to be particularly useful to search for splits in the sequences formed by the upper red giant branch (RGB) and asymptotic giant branch (AGB) stars. We interpret these split sequences as evidence for the presence of MPs. This demonstrates that the J-PLUS survey will have sufficient spatial coverage and spectral resolution to perform a large statistical study of GCs through multi-band photometry in the coming years.Comment: 11 pages, 11 figures. Accepted for publication @ A&

'In vitro' capacitation and acrosome reaction are concomitant with specific changes in mitochondrial activity in boar sperm: evidence for a nucleated mitochondrial activation and for the existence of a capacitation-sensitive subpopulational structure

The main scope of this manuscript is to analyse the dynamics of mitochondrial activity in boar sperm subjected to 'in vitro' capacitation (IVC) and subsequent progesterone-induced 'in vitro' acrosome reaction (IVAR). This was determined after analysis of the rhythm of O(2) consumption and concomitant changes in the mitochondria activity-specific JC-1 staining. Results showed that IVC, and especially IVAR, was concomitant with a peak in O(2) consumption (from 1.61 ± 0.08 nmol O(2)/min/10(7) viable sperm at 0 h of incubation to 2.62 ± 0.12 nmol O(2) /min/10(7) viable sperm after 5 min of IVAR induction). These results were accompanied by parallel changes in the mean intensity of JC-1 staining. Based on JC-1, mitochondrial activation followed a nucleated pattern, with specific, activation starting points at the midpiece from which mitochondrial activation was spread. Moreover, four separate sperm subpopulations were detected following the JC-1 orange-red/green ratio, and the observed changes in the mean JC-1 staining during IVC and IVAR were related to concomitant changes in both the orange-red/green JC-1 ratio and the percentage of sperm included in each subpopulation. All of these results indicate that IVC and the first minutes of IVAR are accompanied by a progressive increase in mitochondrial activity, which reached a peak coincidental with the achievement of IVAR. Moreover, results suggest the presence of separate sperm subpopulations, which show a different mitochondrial sensitivity to IVC and IVAR. Finally, mitochondrial activation, at least under JC-1 staining, seems to originate in concrete nucleation points at the midpiece, thus suggesting thus a well-coordinated pattern in boar-sperm mitochondrial activity modulation

Exome sequencing study in patients with multiple sclerosis reveals variants associated with disease course

BACKGROUND: It remains unclear whether disease course in multiple sclerosis (MS) is influenced by genetic polymorphisms. Here, we aimed to identify genetic variants associated with benign and aggressive disease courses in MS patients. METHODS: MS patients were classified into benign and aggressive phenotypes according to clinical criteria. We performed exome sequencing in a discovery cohort, which included 20 MS patients, 10 with benign and 10 with aggressive disease course, and genotyping in 2 independent validation cohorts. The first validation cohort encompassed 194 MS patients, 107 with benign and 87 with aggressive phenotypes. The second validation cohort comprised 257 patients, of whom 224 patients had benign phenotypes and 33 aggressive disease courses. Brain immunohistochemistries were performed using disease course associated genes antibodies. RESULTS: By means of single-nucleotide polymorphism (SNP) detection and comparison of allele frequencies between patients with benign and aggressive phenotypes, a total of 16 SNPs were selected for validation from the exome sequencing data in the discovery cohort. Meta-analysis of genotyping results in two validation cohorts revealed two polymorphisms, rs28469012 and rs10894768, significantly associated with disease course. SNP rs28469012 is located in CPXM2 (carboxypeptidase X, M14 family, member 2) and was associated with aggressive disease course (uncorrected p value < 0.05). SNP rs10894768, which is positioned in IGSF9B (immunoglobulin superfamily member 9B) was associated with benign phenotype (uncorrected p value < 0.05). In addition, a trend for association with benign phenotype was observed for a third SNP, rs10423927, in NLRP9 (NLR family pyrin domain containing 9). Brain immunohistochemistries in chronic active lesions from MS patients revealed expression of IGSF9B in astrocytes and macrophages/microglial cells, and expression of CPXM2 and NLRP9 restricted to brain macrophages/microglia. CONCLUSIONS: Genetic variants located in CPXM2, IGSF9B, and NLRP9 have the potential to modulate disease course in MS patients and may be used as disease activity biomarkers to identify patients with divergent disease courses. Altogether, the reported results from this study support the influence of genetic factors in MS disease course and may help to better understand the complex molecular mechanisms underlying disease pathogenesis

Estudi sobre la incidencia i la quantificació del dolor postoperatori en la cirurgia de la cesària

La cesària és un procediment de cirurgia major que genera un dolor postoperatori agut, moderat o sever segons els casos. El dolor agut postoperatori es correlaciona amb un augment de la morbilitat postoperatòria. L’objectiu principal de l’estudi fou avaluar la incidència i la quantificació del dolor postoperatori en la cirurgia de cesària a l’Hospital General de Granollers. Durant 2 mesos es van incloure 66 pacients que van ser sotmeses a una cesària. El dolor postoperatori fou alt. Es recomanable realitzar un protocol de tractament del dolor postoperatori en cesàries que sigui eficaç, segur i amb mínims efectes secundaris per a la mare i el nadó

Anestèsia regional per a endarterectomia carotídia. 6 anys d’experiència

Introducció: L’estenosi carotídia causa el 20-25% dels accidents cerebrals vasculars d’origen isquèmic. L’endarterectomia carotídia (EC) es manté com a gold standard en els pacients amb alt grau d’estenosi en l’artèria caròtida interna per a la revascularització carotídia i la prevenció de l’infart cerebral subseqüent. La qüestió de l’anestèsia ideal per la EC ha estat un debat continu des de la primera intervenció. Hi ha evidència científica que les diferents tècniques anestèsiques no són equiparables ni en seguretat ni en qualitat. Així les coses i amb la presumpció que els millor resultats es podien obtenir de l’anestèsia peridural cervical associada a l’anestèsia del plexe cervical superficial, un nou protocol multidisciplinar s’instaura al nostre centre. La nostra opció es basa en la màxima cobertura analgèsica-anestèsica de la combinació de les dues tècniques (anestèsia peridural més plexe cervical superficial), en la major estabilitat hemodinàmica i en la reducció d’estada hospitalària de l’anestèsia regional vs general. El nostre objectiu principal és descriure la qualitat analgèsica-anestèsica de la tècnica. Material i mètodes: Es tracta d’un estudi observacional, descriptiu i retrospectiu. Es van estudiar de forma retrospectiva durant 6 anys (2009-2015) tots els pacients sotmesos a endarterectomia carotídia (EC). Per comparar l’estada hospitalària es van analitzar de forma restrospectiva les dades dels pacients intervinguts entre el 2003 i el 2008, quan la EC es realitzava mitjançant una anestèsia general. Resultats: Es va incloure un total de 88 pacients realitzats sota anestèsia regional i 30 pacients realitzats sota anestèsia general. En les variables principals relacionades amb la qualitat anestèsica es va trobar que un 4,5 % de pacients va requerir complementar amb anestèsic local i 11,2% amb opioids. Cap pacient va desenvolupar un IAM als 30 dies. En un 3,4 % dels casos va ser necessària una conversió a una anestèsia general. Pel que fa a la seguretat de la tècnica anestèsica, no es va trobar cap complicació vital. Es va observar una reducció de l’estada hospitalària de 2 dies en el grup d’anestèsia regional. El cost d’aquests dos dies sumat a l’estalvi en material va representar un estalvi entre 1.688,16 i 2.978,26 euros/pacient. Conclusions: Es va trobar una bona qualitat anestèsica-analgèsica amb l’anestèsia regional, donada la baixa necessitat de infiltració per part del cirurgià, la baixa necessitat d’opioid intraoperatori, la nul·la incidència d’IAM postoperatori i la baixa conversió a una anestèsia general. Hi va haver poques complicacions derivades de la tècnica anestèsica i aquestes van ser lleus. També es va observar una reducció del cost del procediment en el grup regional derivat de la reducció en l’estada hospitalària i de la reducció del consum de material.Introduction: Carotid stenosis is the cause of 20-25% of all vascular cerebral accidents of ischemic origin. Carotid endarterectomy (CE) is still the gold standard in patients with a high degree stenosis of the internal carotid artery for carotid revascularization and subsequent cerebral stroke prevention. Looking for the ideal anaesthetic technique for CE procedures has been a continuous debate since the first intervention. Scientific evidences show that different anaesthetic techniques are not comparable neither in safety nor in quality . Thus, and with the presumption that the best results could be obtained from one cervical epidural anaesthesia combined with anaesthesia of the superficial cervical plexus, a new multidisciplinary protocol was implemented in our setting. Our choice was based on the maximum anaesthetic-analgesic coverage of the combination of the two techniques (peridural anesthesia plus superficial cervical plexus), on a greater hemodynamic stability and on the reduction of hospital stay of the regional vs. general anaesthesia. Our main objective is to describe the analgesic-anesthetic quality of the tecnical. Material and methods: It is an observational, descriptive and retrospective study. We retrospectively analysed during a period of 6 years (2009-2015) all patients undergoing carotid endarterectomy (CE). In order to compare hospital stay, data from patients who underwent the same procedure under general anaesthesia between 2003 and 2008 were analysed retrospectively. Results: A total of 88 patients in the regional anaesthesia group and 30 patients in the general anaesthesia group were included. In terms of anaesthetic quality variables, we observed that 4.5% of patients required complementary local anaesthetic dosage and 11.2% complementary opioids. No patient developed MI at 30 days. In 3.4% of cases, one conversion to general anaesthesia was required. Regarding to anaesthetic technique safety, no vital complications were observed. There was a reduction of hospital length of stay of 2 days in the regional anaesthesia group. The cost of these two days plus material savings represented a saving between 1688.16 and 2978.26 euros per patient. Conclusions: A good anaesthetic-analgesic quality of the regional anesthesia technique was observed due to the following findings: less intraoperative infiltration requirements by the surgeon, less use of intraoperative opioids, no incidence of postoperative MI and less conversion into general anaesthesia. There were only a few complications related to the regional anaesthetic technique without severity. We observed savings in procedure costs due to a shorter hospital stay and to reduced material use in the regional group

Anestèsia regional per a endarterectomia carotídia. 6 anys d’experiència

Introducció: L’estenosi carotídia causa el 20-25% dels accidents cerebrals vasculars d’origen isquèmic. L’endarterectomia carotídia (EC) es manté com a gold standard en els pacients amb alt grau d’estenosi en l’artèria caròtida interna per a la revascularització carotídia i la prevenció de l’infart cerebral subseqüent. La qüestió de l’anestèsia ideal per la EC ha estat un debat continu des de la primera intervenció. Hi ha evidència científica que les diferents tècniques anestèsiques no són equiparables ni en seguretat ni en qualitat. Així les coses i amb la presumpció que els millor resultats es podien obtenir de l’anestèsia peridural cervical associada a l’anestèsia del plexe cervical superficial, un nou protocol multidisciplinar s’instaura al nostre centre. La nostra opció es basa en la màxima cobertura analgèsica-anestèsica de la combinació de les dues tècniques (anestèsia peridural més plexe cervical superficial), en la major estabilitat hemodinàmica i en la reducció d’estada hospitalària de l’anestèsia regional vs general. El nostre objectiu principal és descriure la qualitat analgèsica-anestèsica de la tècnica. Material i mètodes: Es tracta d’un estudi observacional, descriptiu i retrospectiu. Es van estudiar de forma retrospectiva durant 6 anys (2009-2015) tots els pacients sotmesos a endarterectomia carotídia (EC). Per comparar l’estada hospitalària es van analitzar de forma restrospectiva les dades dels pacients intervinguts entre el 2003 i el 2008, quan la EC es realitzava mitjançant una anestèsia general. Resultats: Es va incloure un total de 88 pacients realitzats sota anestèsia regional i 30 pacients realitzats sota anestèsia general. En les variables principals relacionades amb la qualitat anestèsica es va trobar que un 4,5 % de pacients va requerir complementar amb anestèsic local i 11,2% amb opioids. Cap pacient va desenvolupar un IAM als 30 dies. En un 3,4 % dels casos va ser necessària una conversió a una anestèsia general. Pel que fa a la seguretat de la tècnica anestèsica, no es va trobar cap complicació vital. Es va observar una reducció de l’estada hospitalària de 2 dies en el grup d’anestèsia regional. El cost d’aquests dos dies sumat a l’estalvi en material va representar un estalvi entre 1.688,16 i 2.978,26 euros/pacient. Conclusions: Es va trobar una bona qualitat anestèsica-analgèsica amb l’anestèsia regional, donada la baixa necessitat de infiltració per part del cirurgià, la baixa necessitat d’opioid intraoperatori, la nul·la incidència d’IAM postoperatori i la baixa conversió a una anestèsia general. Hi va haver poques complicacions derivades de la tècnica anestèsica i aquestes van ser lleus. També es va observar una reducció del cost del procediment en el grup regional derivat de la reducció en l’estada hospitalària i de la reducció del consum de material.Introduction: Carotid stenosis is the cause of 20-25% of all vascular cerebral accidents of ischemic origin. Carotid endarterectomy (CE) is still the gold standard in patients with a high degree stenosis of the internal carotid artery for carotid revascularization and subsequent cerebral stroke prevention. Looking for the ideal anaesthetic technique for CE procedures has been a continuous debate since the first intervention. Scientific evidences show that different anaesthetic techniques are not comparable neither in safety nor in quality . Thus, and with the presumption that the best results could be obtained from one cervical epidural anaesthesia combined with anaesthesia of the superficial cervical plexus, a new multidisciplinary protocol was implemented in our setting. Our choice was based on the maximum anaesthetic-analgesic coverage of the combination of the two techniques (peridural anesthesia plus superficial cervical plexus), on a greater hemodynamic stability and on the reduction of hospital stay of the regional vs. general anaesthesia. Our main objective is to describe the analgesic-anesthetic quality of the tecnical. Material and methods: It is an observational, descriptive and retrospective study. We retrospectively analysed during a period of 6 years (2009-2015) all patients undergoing carotid endarterectomy (CE). In order to compare hospital stay, data from patients who underwent the same procedure under general anaesthesia between 2003 and 2008 were analysed retrospectively. Results: A total of 88 patients in the regional anaesthesia group and 30 patients in the general anaesthesia group were included. In terms of anaesthetic quality variables, we observed that 4.5% of patients required complementary local anaesthetic dosage and 11.2% complementary opioids. No patient developed MI at 30 days. In 3.4% of cases, one conversion to general anaesthesia was required. Regarding to anaesthetic technique safety, no vital complications were observed. There was a reduction of hospital length of stay of 2 days in the regional anaesthesia group. The cost of these two days plus material savings represented a saving between 1688.16 and 2978.26 euros per patient. Conclusions: A good anaesthetic-analgesic quality of the regional anesthesia technique was observed due to the following findings: less intraoperative infiltration requirements by the surgeon, less use of intraoperative opioids, no incidence of postoperative MI and less conversion into general anaesthesia. There were only a few complications related to the regional anaesthetic technique without severity. We observed savings in procedure costs due to a shorter hospital stay and to reduced material use in the regional group

La inmunoterapia como tratamiento de la atopia en perros y gatos: un update

A abordagem diagnóstica e as subsequentes opções terapêuticas serão fundamentais para uma boa melhoria clínica, também em alergia. Para além das boas práticas, as linhas de orientação específicas para este domínio deverão ser atualizadas e universalizadas, visando uma medicina veterinária de precisão, baseada na evidência, também no domínio da imunoterapia alergénio-específica de animais de companhia