A systematic review of natural products for skin applications: Targeting inflammation, wound healing, and photo-aging.

BACKGROUND: Every day the skin is constantly exposed to several harmful factors that induce oxidative stress. When the cells are incapable to maintain the balance between antioxidant defenses and reactive oxygen species, the skin no longer can keep its integrity and homeostasis. Chronic inflammation, premature skin aging, tissue damage, and immunosuppression are possible consequences induced by sustained exposure to environmental and endogenous reactive oxygen species. Skin immune and non-immune cells together with the microbiome are essential to efficiently trigger skin immune responses to stress. For this reason, an ever-increasing demand for novel molecules capable of modulating immune functions in the skin has risen the level of their development, particularly in the field of natural product-derived molecules. PURPOSE: In this review, we explore different classes of molecules that showed evidence in modulate skin immune responses, as well as their target receptors and signaling pathways. Moreover, we describe the role of polyphenols, polysaccharides, fatty acids, peptides, and probiotics as possible treatments for skin conditions, including wound healing, infection, inflammation, allergies, and premature skin aging. METHODS: Literature was searched, analyzed, and collected using databases, including PubMed, Science Direct, and Google Scholar. The search terms used included "Skin", "wound healing", "natural products", "skin microbiome", "immunomodulation", "anti-inflammatory", "antioxidant", "infection", "UV radiation", "polyphenols", "polysaccharides", "fatty acids", "plant oils", "peptides", "antimicrobial peptides", "probiotics", "atopic dermatitis", "psoriasis", "auto-immunity", "dry skin", "aging", etc., and several combinations of these keywords. RESULTS: Natural products offer different solutions as possible treatments for several skin conditions. Significant antioxidant and anti-inflammatory activities were reported, followed by the ability to modulate immune functions in the skin. Several membrane-bound immune receptors in the skin recognize diverse types of natural-derived molecules, promoting different immune responses that can improve skin conditions. CONCLUSION: Despite the increasing progress in drug discovery, several limiting factors need future clarification. Understanding the safety, biological activities, and precise mechanisms of action is a priority as well as the characterization of the active compounds responsible for that. This review provides directions for future studies in the development of new molecules with important pharmaceutical and cosmeceutical value.

Unsupervised clustering to differentiate rheumatoid arthritis patients based on proteomic signatures.

OBJECTIVE: Patients with rheumatoid arthritis (RA) have different presentations and prognoses. Cluster analysis based on proteomic signatures creates independent phenogroups of patients with different pathophysiological backgrounds. We aimed to identify distinct pathophysiological clusters of RA patients based on circulating proteomic biomarkers. METHOD: This was a cohort study including 399 RA patients. Clustering was performed on 94 circulating proteins (92 CVDII Olink®, high-sensitivity troponin T, and C-reactive protein). Unsupervised clustering was performed using a partitioning cluster algorithm. RESULTS: The clustering algorithm identified two distinct clusters: cluster 1 (n = 223) and cluster 2 (n = 176). Compared with cluster 1, cluster 2 included older patients with a higher burden of comorbidities (cardiovascular and RA related), more erosive and longer RA duration, more dyspnoea and fatigue, walking a shorter distance in the Six-Minute Walk Test, with more severe diastolic dysfunction, and a 4.5-fold higher risk of death or hospitalization for cardiovascular reasons. Tumour necrosis factor (TNF) receptor superfamily-related pathways were mainly responsible for the model's discriminative ability. CONCLUSION: Using unsupervised cluster analysis based on proteomic phenotypes, we identified two clusters of RA patients with distinct biomarkers profiles, clinical characteristics, and different outcomes that could reflect different pathophysiological backgrounds. TNF receptor superfamily-related proteins may be used to distinguish subgroups.

Behavioral and morphological effects of resveratrol and curcumin in rats submitted to doxorubicin-induced cognitive impairment.

Doxorubicin (DOX) is known to cause cognitive impairments in patients submitted to long-term chemotherapy (deficits also known as chemobrain). Therefore, there is an urgent need for therapeutic strategies capable of returning cancer survivors back to their previous quality of life. The present study investigated whether resveratrol (RSV) or curcumin (CUR) administration could affect mnemonic function and brain morphological changes following DOX administration in rats. Male Wistar rats were divided into 4 groups: DOX group (2.5 mg/kg/week for 4 weeks, i.p., plus distilled water for 28 days, oral gavage - OG), DOX + RSV group (DOX, 2.5 mg/kg/week for 4 weeks, i.p., plus RSV, 10 mg/kg/day for 28 days, OG), DOX + CUR group (DOX, 2.5 mg/kg/week for 4 weeks, i.p., plus CUR, 100 mg/kg/day for 28 days, OG) and control (CTR) group (0.9% saline solution weekly for 4 weeks, i.p., plus distilled water for 28 days, OG). Behavioral analyses (open field - OF - and the novel object recognition test - NORT) were performed. Brains were collected and analyzed by hematoxylin-eosin and luxol fast blue staining techniques and by immunohistochemistry for GFAP (glial fibrillary acidic protein) expression in astrocytes and Iba1 (ionized calcium-binding adaptor molecule 1) expression in microglia. DOX-injected rats presented short-term and long-term memory impairments as seen in the NORT at 3 and 24 h after habituation and increased GFAP and Iba1 expression, respectively, in astrocytes and microglia of the frontal cortex, hypothalamus and hippocampus. Such cognitive deficits were prevented by CUR at both periods and by RSV at 24 h. DOX-induced astrogliosis and microgliosis were avoided by RSV and CUR. No signs of demyelination or neuronal loss were found in any group. Thus, CUR and RSV prevented memory loss, astrogliosis and microgliosis induced by DOX monotherapy.

MMP8 increases tongue carcinoma cell-cell adhesion and diminishes migration via cleavage of anti-adhesive FXYD5.

Matrix metalloproteinases (MMPs) modify bioactive factors via selective processing or degradation resulting in tumour-promoting or tumour-suppressive effects, such as those by MMP8 in various cancers. We mapped the substrates of MMP8 to elucidate its previously shown tumour-protective role in oral tongue squamous cell carcinoma (OTSCC). MMP8 overexpressing (+) HSC-3 cells, previously demonstrated to have reduced migration and invasion, showed enhanced cell-cell adhesion. By analysing the secretomes of MMP8 + and control cells with terminal amine isotopic labelling of substrates (TAILS) coupled with liquid chromatography and tandem mass spectrometry (LC-MS/MS), we identified 36 potential substrates of MMP8, including FXYD domain-containing ion transport regulator 5 (FXYD5). An anti-adhesive glycoprotein FXYD5 has been previously shown to predict poor survival in OTSCC. Cleavage of FXYD5 by MMP8 was confirmed using recombinant proteins. Furthermore, we detected a loss of FXYD5 levels on cell membrane of MMP8 + cells, which was rescued by inhibition of the proteolytic activity of MMP8. Silencing (si) FXYD5 increased the cell-cell adhesion of control but not that of MMP8 + cells. siFXYD5 diminished the viability and motility of HSC-3 cells independent of MMP8 and similar effects were seen in another tongue cancer cell line, SCC-25. FXYD5 is a novel substrate of MMP8 and reducing FXYD5 levels either with siRNA or cleavage by MMP8 increases cell adhesion leading to reduced motility. FXYD5 being a known prognostic factor in OTSCC, our findings strengthen its potential as a therapeutic target.

Oncofertility case report: live birth 10 years after oocyte in vitro maturation and zygote cryopreservation.

PURPOSE: This study aims to report a case of urgent fertility preservation in an oncological patient with collection of immature oocytes in the absence of ovarian stimulation that, through in vitro maturation (IVM), followed by ICSI and cryopreservation of zygotes resulted, 10 years later, in the live birth of a healthy baby. METHODS: In September 2008, our clinic performed IVM in a 32-year-old woman diagnosed with a ductal invasive carcinoma with positive estradiol receptors, negative progesterone receptors and positive human epidermal growth factor receptor 2. The retrieval of immature oocytes was performed in the absence of ovarian stimulation after a simple mastectomy and prior to any chemotherapy treatment. The compact cumulus-oocyte complexes (COCs) collected were placed in Lag medium for 2 h, followed by incubation in IVM medium, supplemented with heat inactivated patient serum, recombinant FSH, and recombinant LH. After 30 h in culture, cumulus cells were removed, the metaphase II oocytes were microinjected, and the zygotes obtained were cryopreserved. In 2017, the zygotes were thawed and cultured until day 3. One embryo was transferred and the other cryopreserved. RESULTS: Four compact COCs were collected and subjected to IVM. Two oocytes reached metaphase II and were microinjected. Two zygotes were obtained and were cryopreserved at the two pronuclear stage. Approximately 9 years later, the two zygotes were thawed and cultured until day 3. An embryo with 10 cells was transferred and implanted, resulting in the birth of a healthy baby. CONCLUSIONS: In cases where urgency to start adjuvant therapy requires immediate oocyte collection, IVM may be the only option to obtain fully competent mature oocytes allowing for effective preservation of the reproductive potential.

Greenhouse gas balance and carbon footprint of pasture-based beef cattle production systems in the tropical region (Atlantic Forest biome).

The production of beef cattle in the Atlantic Forest biome mostly takes place in pastoral production systems. There are millions of hectares covered with pastures in this biome, including degraded pasture (DP), and only small area of the original Atlantic Forest has been preserved in tropics, implying that actions must be taken by the livestock sector to improve sustainability. Intensification makes it possible to produce the same amount, or more beef, in a smaller area; however, the environmental impacts must be assessed. Regarding climate change, the C dynamics is essential to define which beef cattle systems are sustainable. The objectives of this study were to investigate the C balance (t CO2e./ha per year), the intensity of C emission (kg CO2e./kg BW or carcass) and the C footprint (t CO2e./ha per year) of pasture-based beef cattle production systems, inside the farm gate and considering the inputs. The results were used to calculate the number of trees to be planted in beef cattle production systems to mitigate greenhouse gas (GHG) emissions. The GHG emission and C balance, for 2 years, were calculated based on the global warming potential (GWP) of AR4 and GWP of AR5. Forty-eight steers were allotted to four grazing systems: DP, irrigated high stocking rate pasture (IHS), rainfed high stocking rate pasture (RHS) and rainfed medium stocking rate pasture (RMS). The rainfed systems (RHS and RMS) presented the lowest C footprints (-1.22 and 0.45 t CO2e./ha per year, respectively), with C credits to RMS when using the GWP of AR4. The IHS system showed less favorable results for C footprint (-15.71 t CO2e./ha per year), but results were better when emissions were expressed in relation to the annual BW gain (-10.21 kg CO2e./kg BW) because of its higher yield. Although the DP system had an intermediate result for C footprint (-6.23 t CO2e./ha per year), the result was the worst (-30.21 CO2e./kg BW) when the index was expressed in relation to the annual BW gain, because in addition to GHG emissions from the animals in the system there were also losses in the annual rate of C sequestration. Notably, the intensification in pasture management had a land-saving effect (3.63 ha for IHS, 1.90 for RHS and 1.19 for RMS), contributing to the preservation of the tropical forest.

Expressão e caracterização da glicoproteína D do herpesvírus equídeo 1 em Pichia pastoris / Expression and characterization of equid herpesvirus 1 glycoprotein D in Pichia pastoris

O herpesvírus equídeo 1 (EHV-1) apresenta distribuição mundial e causa graves prejuízos à equideocultura. É agente de surtos de doença respiratória, reprodutiva e neurológica, em equídeos jovens e adultos. A glicoproteína D (gD) do envelope viral é essencial para ligação e penetração em células permissivas e direcionamento do sistema imunológico do hospedeiro, induz respostas imunes humorais e celulares, sendo um antígeno apropriado para ser utilizado em vacinas e imunodiagnóstico. O objetivo deste trabalho foi expressar e caracterizar a gD do EHV-1 em Pichia pastoris para posterior utilização como antígeno em técnicas de imunodiagnóstico e formulação de vacinas recombinantes. Uma sequência de DNA que codifica uma forma truncada da gDEHV-1 foi clonada no vetor pPICZαA de expressão em P. pastoris. Obteve-se uma proteína de ~41 kDa, como esperado. A proteína apresentou glicosilação entre 4 kDa e 16 kDa, demonstrada por deglicosilação enzimática. A proteína recombinante foi caracterizada antigenicamente e imunogenicamente por Western blot, utilizando-se anticorpos policlonais equinos anti-EHV-1, e por ELISA indireto em modelo murino, demonstrando que a gD recombinante manteve epítopos similares aos da proteína nativa. Esses resultados sugerem que a gDEHV-1 é um antígeno promissor para uso como imunobiológico no controle do EHV-1.(AU)

Equine herpesvirus 1 (EHV-1) has a worldwide distribution and causes serious damage to horse breeding. It is an agent of respiratory, reproductive and neurological disease outbreaks in young and adult equids. Viral envelope glycoprotein D (gD) is essential for binding and penetration into permissive cells and targeting the host immune system, inducing humoral and cellular immune responses, and is an appropriate antigen for use in vaccines and immunodiagnostics. The objective of this work was to express in Pichia pastoris and to characterize EHV-1 gD for later use as an antigen in immunodiagnostic techniques and formulation of recombinant vaccines. A DNA sequence encoding a truncated form of gDEHV-1 has been cloned into the P. pastoris expression vector pPICZαA. A protein of ~41 kDa was obtained as expected. The protein presented glycosylation between 4 kDa and 16 kDa, demonstrated by enzymatic deglycosylation. The recombinant protein was antigenically and immunogenically characterized by Western blot using equine polyclonal anti-EHV-1 antibodies, and by indirect ELISA in a murine model, demonstrating that the recombinant gD maintained epitopes similar to those of the native protein. These results suggest that gDEHV-1 is a promising antigen for use as an immunobiological in the control of EHV-1.(AU)

Prenatal LPS induces sickness behaviour and decreases maternal and predatory behaviours after an LPS challenge.

Purpose: The influence of a challenge dose of lipopolysaccharide (LPS) on the behavioural selection between maternal (MB) and predatory behaviours (PB) of female rats prenatally treated with the same endotoxin or saline solution (F1 generation) were studied.Material and methods: Thus, in adult age, these female rats were mated and, at lactation days 5 or 6, the following groups were formed: (1) LPS + LPS group-female rats prenatally treated with LPS and received an LPS challenge dose; (2) S + LPS group-female rats prenatally treated with saline solution and received a challenge LPS dose (3) S + S group-females rats prenatally treated with saline which received a saline injection. MB, PB to cockroaches, exploratory behaviour, periaqueductal grey (PAG) expression of the astrocytic biomarker glial fibrillary acidic protein (GFAP), and corticosterone and TNF-alpha serum levels were evaluated.Results: Showed that: (1) relative to the S + S group, the LPS + S group showed decreased MB and slightly increased PB, without inducing sickness behaviour; (2) the LPS + LPS group showed decreased MB but few effects on PB; (3) there was increased sickness behaviour associated with increased TNF-alpha serum levels in the LPS + LPS group; (4) a significant increase in GFAP expression was observed in both LPS groups, which was greater in the LPS + LPS group and (5) no differences in the corticosterone of all groups.Conclusions: Prenatal LPS impaired the switch from MB to PB in female rats of the LPS + LPS group by increased sickness behaviour as well as an increase in plasmatic TNF-alpha levels inducing PAG astrogliosis.

Health economic assessment of a shift to active transport.

Active transportation (walking or cycling) as a substitute for car trips still represents a small percentage of all daily travels in many European cities. This study aimed to estimate the health and economic co-benefits for the adult population of modal shift from driving to active travel in urban environments. Three scenarios were modelled for the case study, the city of Porto, Portugal, by comparing travel patterns of 2013 to hypothetical scenarios of modal shifts from driving to active transport, namely: i) SC1 - conservative scenario, with a change of 5% from driving to cycling and 10% from driving to walking; ii) SC2 - moderate scenario, with a shift of 10% and 15%, respectively; and iii) SC3 - optimistic scenario, with a shift of 15% and 20%, respectively. The mortality risk reduction for five health outcomes (colon and breast cancers, diabetes, ischemic heart disease, cerebrovascular disease) was assessed, including an estimation of traffic injury and air pollution exposure risks. Results were presented in Disability-Adjusted Life Years (DALYs) avoided. Economic valuation for each scenario was performed using a Willingness-to-Pay approach for morbimortality and a Cost of Illness approach for 2013 hospitalizations and work absenteeism. Significant health benefits were found in all modelled scenarios, ranging from 1657 (16%) to 2881 (28%) DALYs avoided. Total costs averted ranged from €3894 to €6769 million through the scenarios. Cardio and cerebrovascular diseases mortality presented the largest benefit, accounting for about 3/4 of all avoidable DALYs in all scenarios. Reductions in CO2 and PM10 emissions were calculated, showing a decrease from 31.6 to 73.7 kt of CO2 and 7 to 16 t for PM10, respectively. A modal shift towards active transportation could lead to significant health and economic benefits, indicating that the evaluation of health impacts should be included in the analysis of active transport interventions.

Impact of the systematic use of the Gugging Swallowing Screen in patients with acute ischaemic stroke.

BACKGROUND AND PURPOSE: Post-stroke dysphagia occurs in up to three quarters of patients with acute stroke and is associated with a higher risk of respiratory infections and poor outcome. Systematic screening of dysphagia in the acute stroke unit is essential to identify patients at risk of aspiration and to provide dietary recommendations. Our study aimed to assess the impact of the systematic application of the Gugging Swallowing Screen (GUSS) in patients with acute ischaemic stroke. METHODS: This was a retrospective study of consecutive patients with acute ischaemic stroke admitted to an acute stroke unit in two time periods: pre-GUSS (February 2014-July 2015), when the 10-mL water-swallowing test was systematically administered, and GUSS (August 2015-October 2016), when the GUSS test was systematically administered. Groups were compared with regard to baseline and stroke characteristics, and the occurrence of stroke-associated pneumonia (SAP), in-hospital death and 3-month outcome. RESULTS: Of the 344 patients who were included in the study (median age 71 years), 51.7% were male with a median National Institutes of Health Stroke Scale score of 11. A total of 204 patients were included during the pre-GUSS period and 140 during the GUSS period. Patients in the GUSS period more frequently had diabetes and partial anterior circulation syndromes, and were more frequently treated with thrombectomy. There was no difference in the occurrence of SAP between the two groups (pre-GUSS, 12.5%; GUSS, 15.1%; P = 0.490) and no differences were found concerning in-hospital mortality (P = 0.996), 3-month functional independence (P = 0.647) or 3-month mortality (P = 0.598). CONCLUSIONS: The systematic administration of GUSS in a population of patients with acute ischaemic stroke did not reduce the occurrence of SAP, mortality or 3-month functional dependence when compared with the systematic administration of the 10-mL water-swallowing test.

Monensin and a blend of castor oil and cashew nut shell liquid used in a high-concentrate diet abruptly fed to Nellore cattle.

Monensin and functional oils (FO) were supplemented to a high-concentrate diet abruptly fed to 12 ruminally cannulated Zebu steers to study their effects on rumen fermentation, blood metabolites, and , , and relative population. A randomized complete block design with repeated measures over time within 2 experimental periods of 21 d each was used. Treatments were a control (CTR; with no additives), FO (included at 400 mg/kg), and monensin included at 30 mg/kg (M30) or 40 mg/kg (M40). All steers were fed the same high-concentrate basal diet, which consisted of 92.25% concentrate. The first 60 h after transition showed a treatment and hour interaction for ruminal propionate proportion ( = 0.028), and no change in acetate molar proportion ( = 0.633), rumen pH ( = 0.370), and time the rumen pH remained below 5.6 ( = 0.242) were observed. The acetate:propionate ratio decreased ( = 0.020) when monensin was fed in both concentrations (2.30 for the M30 treatment and 2.32 for the M40 treatment) compared with when the CTR was fed (2.85), without being different when the FO (2.71) treatment was fed. Only the M30 treatment did not show pH below 5.2 (P=0.047) over the 60 h after the abrupt transition. Within the entire period, DMI ( = 0.008) and mean ruminal pH ( = 0.040) as well as molar proportions of propionate ( = 0.034) and valerate ( = 0.031) had significant interactions between treatment and day. Total VFA concentration was greater ( = 0.017) for the M30 (117.36 m) and CTR treatments (115.77 m) compared with the M40 treatment (105.02 m), without being different for the FO treatment (111.55 m). Treatments did not change feed behavior parameters. Blood HCO ( = 0.006) and total carbon dioxide ( = 0.003) were greater for the M30 (27.8 and 29.3 mmol/L, respectively) and FO treatments (28.3 and 29.7 mmol/L, respectively) compared with the CTR treatment (25.7 and 26.9 mmol/L, respectively). ( < 0.0001) and ( < 0.0001) decreased their population throughout days, whereas ( = 0.026) increased its population. Independent of ciliated protozoa genera, the greatest ( < 0.0001) protozoa counts were observed for the CTR treatment (52.7 × 10/mL), intermediate for the FO treatment (35.3 x10/mL), and least for steers fed monensin in both concentrations (15 × 10/mL for the M30 treatment and 14 × 10/mL for the M40 treatment). Feed additives had different effects to reduce the subacute acidosis. The use of the FO and M40 treatments did not change most of the rumen fermentation variables, especially in the first week after abrupt transition, when the M30 treatment provided higher protection against acidosis.

Human leukocyte antigen distribution and genomic ancestry in Brazilian patients with sickle cell disease.

Hematopoietic stem-cell transplantation (HSCT) is currently the only established curative treatment for sickle cell disease (SCD), but is limited by donor availability. Ethnicity is thought to have an impact on the complications experienced by patients that undergo HSCT and on the likelihood of identifying an human leukocyte antigen (HLA) matched donor. In the present study, we investigated the genomic ancestry and the distribution of HLA allele groups in Brazilian patients with SCD, compared these HLA profiles to worldwide populations and evaluate the availability of HLA-matched donors. A broad intercontinental admixture of patients with SCD was observed, with African ancestry ranging from 6.7% to 93.4%. In a dendrogram based on HLA frequencies, Brazilian patients with SCD were included in a branch containing only populations with a significant African component. Among the 126 patients evaluated, 10 (8%) found a HLA-matched unrelated donor in a database of 18 134 donors. Self-reported white, brown and black matched donors were identified, and no significant difference in the percentage of compatible donors was observed between these ethnic groups. Our results show that Brazilian patients with SCD are very admixed, indicating that this group is a promising target for admixture mapping of genes involved in complications after HSCT. Additional studies may help to clarify the impact of the genetic diversity and admixture of these patients on the donor availability.

Antimicrobial and immunomodulatory activity of host defense peptides, clavanins and LL-37, in vitro: An endodontic perspective.

Endodontic treatment is mainly based on root canal disinfection and its failure may be motivated by microbial resistance. Endodontic therapy can be benefitted by host defense peptides (HDPs), which are multifunctional molecules that act against persistent infection and inflammation. This study aimed to evaluate the antimicrobial, cytotoxic and immunomodulatory activity of several HDPs, namely clavanin A, clavanin A modified (MO) and LL-37, compared to intracanal medication Ca(OH)2. HDPs and Ca(OH)2 were evaluated by: (1) antimicrobial assays against Candida albicans and Enterococcus faecalis, (2) cytotoxicity assays and (3) cytokine tumor necrosis factor (TNF)-α, monocyte chemoattractant protein (MCP)-1, interleukin (IL)-1α, IL-6, IL-10 and IL-12 and nitric oxide (NO) production by RAW 264.7 cells incubated with or without heat-killed (HK) C. albicans or E. faecalis combined or not with interferon-γ. The minimum inhibitory concentration (MIC) was established only for E. faecalis (LL-37, 57µM). Considering cytotoxicity, clavanin MO was able to reduce cell viability in many groups and demonstrated lowest LC50. The Ca(OH)2 up-regulated the production of MCP-1, TNF-α, IL-12 and IL-6 and down-regulated IL-1α, IL-10 and NO. Clavanins up-regulated the TNF-α and NO and down-regulated IL-10 production. LL-37 demonstrated up-regulation of IL-6 and TNF-α production and down-regulation in IL-10 and NO production. In conclusion, LL-37 demonstrated better antibacterial potential. In addition, Ca(OH)2 demonstrated a proinflammatory response, while the HDPs modulated the inflammatory response from non-interference with the active cytokines in the osteoclastogenesis process, probably promoting the health of periradicular tissues.

Membrane-enriched proteome changes and prion protein expression during neural differentiation and in neuroblastoma cells.

BACKGROUND: The function of the prion protein, involved in the so-called prion diseases, remains a subject of intense debate and the possibility that it works as a pleiotropic protein through the interaction with multiple membrane proteins is somehow supported by recent reports. Therefore, the use of proteomic and bioinformatics combined to uncover cellular processes occurring together with changes in the expression of the prion protein may provide further insight into the putative pleiotropic role of the prion protein. RESULTS: This study assessed the membrane-enriched proteome changes accompanying alterations in the expression of the prion protein. A 2D-DIGE approach was applied to two cell lines after prefractionation towards the membrane protein subset: an embryonic stem cell line and the PK1 subline of neuroblastoma cells which efficiently propagates prion infection. Several proteins were differentially abundant with the increased expression of the prion protein during neural differentiation of embryonic stem cells and with the knockdown of the prion protein in PK1 cells. The identity of around 20% of the differentially abundant proteins was obtained by tandem MS. The catalytic subunit A of succinate dehydrogenase, a key enzyme for the aerobic energy metabolism and redox homeostasis, showed a similar abundance trend as the prion protein in both proteomic experiments. A gene ontology analysis revealed "myelin sheath", "organelle membrane" and "focal adhesion" associated proteins as the main cellular components, and "protein folding" and "ATPase activity" as the biological processes enriched in the first set of differentially abundant proteins. The known interactome of these differentially abundant proteins was customized to reveal four interactors with the prion protein, including two heat shock proteins and a protein disulfide isomerase. CONCLUSIONS: Overall, our study shows that expression of the prion protein occurs concomitantly with changes in chaperone activity and cell-redox homeostasis, emphasizing the functional link between these cellular processes and the prion protein.

Physiological responses of reared sea bream (Sparus aurata Linnaeus, 1758) to an Amyloodinium ocellatum outbreak.

Amyloodiniosis represents a major bottleneck for semi-intensive aquaculture production in Southern Europe, causing extremely high mortalities. Amyloodinium ocellatum is a parasitic dinoflagellate that can infest almost all fish, crustacean and bivalves that live within its ecological range. Fish mortalities are usually attributed to anoxia, associated with serious gill hyperplasia, inflammation, haemorrhage and necrosis in heavy infestations; or with osmoregulatory impairment and secondary microbial infections due to severe epithelial damage in mild infestation. However, physiological information about the host responses to A. ocellatum infestation is scarce. In this work, we analysed the proteome of gilthead sea bream (Sparus aurata) plasma and relate it with haematological and immunological indicators, in order to enlighten the different physiological responses when exposed to an A. ocellatum outbreak. Using 2D-DIGE, immunological and haematological analysis and in response to the A. ocellatum contamination we have identified several proteins associated with acute-phase response, inflammation, lipid transport, homoeostasis, and osmoregulation, wound healing, neoplasia and iron transport. Overall, this preliminary study revealed that amyloodiniosis affects some fish functional pathways as revealed by the changes in the plasma proteome of S. aurata, and that the innate immunological system is not activated in the presence of the parasite.

Leptin receptor expression and Gln223Arg polymorphism as prognostic markers in oral and oropharyngeal cancer.

The leptin gene product is released into the blood stream, passes through the blood-brain barrier, and finds the leptin receptor (LEPR) in the central nervous system. This hormone regulates food intake, hematopoiesis, inflammation, immunity, differentiation, and cell proliferation. The LEPR Gln223Arg polymorphism has been reported to alter receptor function and expression, both of which have been related with prognostics in several tumor types. Furthermore, several studies have shown a relationship between the Gln223Arg polymorphism and tumor development, and its role in oral and oropharyngeal squamous cell carcinoma is now well understood. In this study, 315 DNA samples were used for LEPR Gln223Arg genotyping and 87 primary oral and oropharyngeal squamous cell carcinomas were used for immunohistochemical expression analysis, such that a relationship between these and tumor development and prognosis could be established. Homozygous LEPR Arg223 was found to be associated with a 2-fold reduction in oral and oropharyngeal cancer risk. In contrast, the presence of the Arg223 allele in tumors was associated with worse disease-free and disease-specific survival. Low LEPR expression was found to be an independent risk factor, increasing the risk for lymph node metastasis 4-fold. In conclusion, the Gln223Arg polymorphism and LEPR expression might be valuable markers for oral and oropharyngeal cancer, suggesting that LEPR might serve as a potential target for future therapies.

Effects of feeding a spray-dried multivalent polyclonal antibody preparation on feedlot performance, feeding behavior, carcass characteristics, rumenitis, and blood gas profile of Brangus and Nellore yearling bulls.

The objective of this study was to evaluate the effects of replacing monensin (MON) with a spray-dried multivalent polyclonal antibody preparation (PAP) against several ruminal microorganisms on feedlot performance, carcass characteristics, feeding behavior, blood gas profile, and the rumenitis incidence of Brangus and Nellore yearling bulls. The study was designed as a completely randomized design with a 2 × 2 factorial arrangement, replicated 6 times (4 bulls per pen and a total of 24 pens), in which bulls ( = 48) of each biotype were fed diets containing either MON fed at 300 mg/d or PAP fed at 3 g/d. No significant feed additive main effects were observed for ADG ( = 0.27), G:F ( = 0.28), HCW ( = 0.99), or dressing percentage ( = 0.80). However, bulls receiving PAP had greater DMI ( = 0.02) and larger ( = 0.02) final LM area as well as greater ( < 0.01) blood concentrations of bicarbonate and base excess in the extracellular fluid than bulls receiving MON. Brangus bulls had greater ( < 0.01) ADG and DMI expressed in kilograms, final BW, heavier HCW, and larger initial and final LM area than Nellore bulls. However, Nellore bulls had greater daily DMI fluctuation ( < 0.01), expressed as a percentage, and greater incidence of rumenitis ( = 0.05) than Brangus bulls. In addition, Brangus bulls had greater ( < 0.01) DMI per meal and also presented lower ( < 0.01) DM and NDF rumination rates when compared with Nellore bulls. Significant interactions ( < 0.05) between biotype and feed additive were observed for SFA, unsaturated fatty acids (UFA), MUFA, and PUFA concentrations in adipose tissues. When Nellore bulls were fed PAP, fat had greater ( < 0.05) SFA and PUFA contents but less ( < 0.01) UFA and MUFA than Nellore bulls receiving MON. For Brangus bulls, MON led to greater ( < 0.05) SFA and PUFA and less ( < 0.05) UFA and MUFA than Brangus bulls fed PAP. Feeding a spray-dried PAP led to similar feedlot performance compared with that when feeding MON. Spray-dried PAP might provide a new technology alternative to ionophores.

Surgical management of cervical spine instability in rheumatoid arthritis patients.

AIM: Cross-sectional study that aims to evaluate the results of cervical spine surgeries due to rheumatoid arthritis (RA) instability, between January of 2000 and of 2012 in a main Portuguese centre. METHODS: Patients followed on Rheumatology submitted to cervical spine fusion due to atlantoaxial (AAI), sub-axial (SAI) or cranio-cervical (CCI) instabilities between 2000-2012 were included. Information about the surgical procedure and associated complications was gathered and imagiologic and clinical indexes before and after surgery (as anterior and posterior atlanto-axial interval and Ranawat index) were evaluated and compared using adequate statistics. RESULTS: Forty-five patients with RA were included: 25 with AAI, 13 with CCI and 7 with SAI. Ten AAI and 4 CCI patients were submitted to wiring stabilization techniques; 15 AAI and 9 CCI patients to rigid ones; and in all patients with SAI an anterior cervical arthrodesis was chosen. There is a significant increase in PADI and a decrease in AADI in the postoperative evaluation (p<0,05) that only remains significant when rigid systems were used. After surgery the Ranawat index decreased (p<0,05) and no patient showed a deterioration of neurological condition. The complication rate was of 23,1%, with 5 mal-unions. CONCLUSION: Surgical management of cervical column instabilities in patients with rheumatoid arthritis seems to be a safe procedure, with a high rate of neurologic improvement. Rigid techniques seem to lead to a better imagiological improvement when compared to wiring ones.