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Comparative assessment of probiotics and monensin in the prophylaxis of acute ruminal lactic acidosis in sheep
No full textAbstract\ud
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Background\ud
Acute ruminal lactic acidosis (ARLA) is a major nutritional and metabolic disorder usually characterized by excessive or non-adapted intake of diets rich in nonstructural carbohydrates. Feed additives that regulate the ruminal environment have been used to prevent ARLA, such as ionophores and, more recently, yeast culture. Thus, we aimed to compare the efficacy of a yeast-based culture (Saccharomyces cerevisiae) with that of monensin sodium in the prevention of ARLA in sheep. Eighteen male, crossbred, rumen-cannulated sheep were randomly distributed into three groups of six animals: control, yeast culture and monensin. Thirty days after the start of supplementation with yeast culture (4 × 109 cfu/animal/day of S. cerevisiae) and monensin (33 mg/kg of total dry matter intake), 15 g/kg BW of sucrose was administered directly into the rumen of the animals to induce ARLA. Samples of blood and ruminal fluid were collected at the following time points: at baseline (T0 h) immediately before the induction of ARLA; 6 h (T6 h); 12 h (T12 h); 18 h (T18 h); 24 h (T24 h); 36 h (T36 h); and 48 h (T48 h) after ARLA induction.\ud
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Results\ud
Ruminal pH was higher in monensin group at T12 h and in yeast culture group at T36 h when compared to control group. Lower values of L-Lactate were found at yeast culture group at T24 h and T36 h. Monensin showed prophylactic effect by decreasing the rate of ruminal pH decline and occasionally reducing ruminal acidosis, whereas probiotics resulted in less accumulation of lactic acid in the rumen and a lower degree of systemic acidosis.\ud
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Conclusion\ud
The use of yeast culture can be beneficial in the prevention and treatment of ARLA in sheep, because it can effectively reduce the accumulation of lactic acid, and thereby increase ruminal pH and reduce ruminal osmolarity. On the other hand, monensin showed prophylactic effect by decreasing the rate of ruminal pH decline and occasionally reducing ruminal acidosis, however, it did not directly prevent these conditions.Financial support for this study was provided by the Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)
FAM3B/PANDER inhibits cell death and increases prostate tumor growth by modulating the expression of Bcl-2 and Bcl-XL cell survival genes
No full textAbstract\ud
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Background\ud
FAM3B/PANDER is a novel cytokine-like protein that induces apoptosis in insulin-secreting beta-cells. Since in silico data revealed that FAM3B can be expressed in prostate tumors, we evaluated the putative role of this cytokine in prostate tumor progression.\ud
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Methods\ud
FAM3B expression was analyzed by quantitative PCR in tumor tissue clinical samples and prostate tumor cell lines. Culture growth and viability of DU145 cell line were evaluated after treatment with either exogenous FAM3B protein obtained from conditioned media (CM) of 293 T cells overexpressing FAM3B or a recombinant FAM3B protein produced in a bacterial host. DU145 cells overexpressing FAM3B protein were produced by lentiviral-mediated transduction of full-length FAM3B cDNA. Cell viability and apoptosis were analyzed in DU145/FAM3B cells after treatment with several cell death inducers, such as TNF-alpha, staurosporine, etoposide, camptothecin, and serum starvation conditions. Anchorage-independent growth in soft agarose assay was used to evaluate in vitro tumorigenicity. In vivo tumorigenicity and invasiveness were evaluated by tumor xenograft growth in nude mice.\ud
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Results\ud
We observed an increase in FAM3B expression in prostate tumor samples when compared to normal tissues. DU145 cell viability and survival increased after exogenous treatment with recombinant FAM3B protein or FAM3B-secreted protein. Overexpression of FAM3B in DU145 cells promoted inhibition of DNA fragmentation and phosphatidylserine externalization in a time and dose-dependent fashion, upon apoptosis triggered by TNF-alpha. These events were accompanied by increased gene expression of anti-apoptotic Bcl-2 and Bcl-XL, decreased expression of pro-apoptotic Bax and diminished caspase-3, −8 and −9 proteolytic activities. Furthermore, inhibition of Bcl-2 anti-apoptotic family proteins with small molecules antagonists decreases protective effects of FAM3B in DU145 cells. When compared to the respective controls, cells overexpressing FAM3B displayed a decreased anchorage- independent growth in vitro and increased tumor growth in xenografted nude mice. The immunohistochemistry analysis of tumor xenografts revealed a similar anti-apoptotic phenotype displayed by FAM3B-overexpressing tumor cells.\ud
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Conclusions\ud
Taken together, by activating pro-survival mechanisms FAM3B overexpression contributes to increased resistance to cell death and tumor growth in nude mice, highlighting a putative role for this cytokine in prostate cancer progression.This work was partially supported by the São Paulo State Research Foundation (FAPESP. Proc 2007/04513–1) and the Brazilian National Council for Scientific and Technological Development (CNPq. Proc. 2011/486048). The funding agents had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript. We finalized this work despite the few resources available and the financial crisis provoked by the austerity policies of the Brazilian Government that directly affect the public university budget, the R&D perspectives and the formation of new scientists in our country
Nutritional risk in outpatients of a geriatric neuropsychiatry clinic
No full textAbstract\ud
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Background\ud
There are few published studies investigating the nutritional status of elderly individuals with psychiatric disorders. This study aims to describe the nutritional status of elderly patients from a neuropsychiatry outpatient clinic, investigating their nutritional status according to the type of psychiatric diagnostic, specifically Alzheimer’s disease (AD), depression (DEP), and other types of dementia (OTD).\ud
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Methods\ud
Elderly individuals from two outpatient clinics (n = 217) were evaluated for nutritional risk (using a validated tool) and for some anthropometric and biochemical measurements.\ud
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Results\ud
We found a high prevalence of nutritional risk in the sample (about 60%). The major dietary problems found were low daily meal frequency and the low daily intake of water. Biochemical analyses indicated blood glucose and total cholesterol to be above the reference values, while hemoglobin and vitamin D were below the reference values. Anthropometric measurements did not differ between the groups with different psychiatric diagnostics, except for calf circumference, which showed to be lower in the OTD group (p = 0.006).\ud
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Conclusions\ud
Although we found a high prevalence of nutritional risk, the differences between the diagnostic groups were very subtle. The nutritional risk is certainly associated with inadequate eating habits. It is necessary to seek strategies to improve the diet and other lifestyle factors in geriatric neuropsychiatry, in order to mitigate the negative outcomes brought about by the diseases
Techno-economic analysis of the industrial production of a low-cost enzyme using E. coli: the case of recombinant β-glucosidase
No full textAbstract\ud
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Background\ud
The enzymatic conversion of lignocellulosic biomass into fermentable sugars is a promising approach for producing renewable fuels and chemicals. However, the cost and efficiency of the fungal enzyme cocktails that are normally employed in these processes remain a significant bottleneck. A potential route to increase hydrolysis yields and thereby reduce the hydrolysis costs would be to supplement the fungal enzymes with their lacking enzymatic activities, such as β-glucosidase. In this context, it is not clear from the literature whether recombinant E. coli could be a cost-effective platform for the production of some of these low-value enzymes, especially in the case of on-site production. Here, we present a conceptual design and techno-economic evaluation of the production of a low-cost industrial enzyme using recombinant E. coli.\ud
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Results\ud
In a simulated baseline scenario for β-glucosidase demand in a hypothetical second-generation ethanol (2G) plant in Brazil, we found that the production cost (316 US$/kg) was higher than what is commonly assumed in the literature for fungal enzymes, owing especially to the facility-dependent costs (45%) and to consumables (23%) and raw materials (25%). Sensitivity analyses of process scale, inoculation volume, and volumetric productivity indicated that optimized conditions may promote a dramatic reduction in enzyme cost and also revealed the most relevant factors affecting production costs.\ud
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Conclusions\ud
Despite the considerable technical and economic uncertainties that surround 2G ethanol and the large-scale production of low-cost recombinant enzymes, this work sheds light on some relevant questions and supports future studies in this field. In particular, we conclude that process optimization, on many fronts, may strongly reduce the costs of E. coli recombinant enzymes, in the context of tailor-made enzymatic cocktails for 2G ethanol production.This work received fnancial support from Fundação de Amparo à Pesquisa\ud
do Estado de São Paulo—FAPESP (São Paulo, Brazil, Grants 2014/13974-6\ud
and 2010/08089-2) and Conselho Nacional de Desenvolvimento Científco e\ud
Tecnológico—CNPq (Brazil, Grant 400803/2013-5)
Transcriptome and secretome analysis of Aspergillus fumigatus in the presence of sugarcane bagasse
No full textAbstract\ud
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Background\ud
Sugarcane bagasse has been proposed as a lignocellulosic residue for second-generation ethanol (2G) produced by breaking down biomass into fermentable sugars. The enzymatic cocktails for biomass degradation are mostly produced by fungi, but low cost and high efficiency can consolidate 2G technologies. A. fumigatus plays an important role in plant biomass degradation capabilities and recycling. To gain more insight into the divergence in gene expression during steam-exploded bagasse (SEB) breakdown, this study profiled the transcriptome of A. fumigatus by RNA sequencing to compare transcriptional profiles of A. fumigatus grown on media containing SEB or fructose as the sole carbon source. Secretome analysis was also performed using SDS-PAGE and LC-MS/MS.\ud
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Results\ud
The maximum activities of cellulases (0.032 U mL-1), endo-1,4-β--xylanase (10.82 U mL-1) and endo-1,3-β glucanases (0.77 U mL-1) showed that functional CAZymes (carbohydrate-active enzymes) were secreted in the SEB culture conditions. Correlations between transcriptome and secretome data identified several CAZymes in A. fumigatus. Particular attention was given to CAZymes related to lignocellulose degradation and sugar transporters. Genes encoding glycoside hydrolase classes commonly expressed during the breakdown of cellulose, such as GH-5, 6, 7, 43, 45, and hemicellulose, such as GH-2, 10, 11, 30, 43, were found to be highly expressed in SEB conditions. Lytic polysaccharide monooxygenases (LPMO) classified as auxiliary activity families AA9 (GH61), CE (1, 4, 8, 15, 16), PL (1, 3, 4, 20) and GT (1, 2, 4, 8, 20, 35, 48) were also differentially expressed in this condition. Similarly, the most important enzymes related to biomass degradation, including endoxylanases, xyloglucanases, β-xylosidases, LPMOs, α-arabinofuranosidases, cellobiohydrolases, endoglucanases and β-glucosidases, were also identified in the secretome.\ud
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Conclusions\ud
This is the first report of a transcriptome and secretome experiment of Aspergillus fumigatus in the degradation of pretreated sugarcane bagasse. The results suggest that this strain employs important strategies for this complex degradation process. It was possible to identify a set of genes and proteins that might be applied in several biotechnology fields. This knowledge can be exploited for the improvement of 2G ethanol production by the rational design of enzymatic cocktails.This study was supported by Fundação de Amparo À Pesquisa do Estado de\ud
São Paulo (FAPESP, 2014/10466–0). AVB and LEG were funded by scholarships\ud
granted by Conselho Nacional de Desenvolvimento Científico e Tecnológico\ud
(CNPq). PFG was funded by a scholarship granted by Coordenação de\ud
Aperfeiçoamento de Pessoal de Ensino Superior (CAPES)
Staying in the labor force among patients with rheumatoid arthritis and associated factors in Southern Brazil
No full textAbstract
Background
Rheumatoid arthritis primarily affects the working-age population and may cause key functional and work limitations. As the disease progresses, individuals become increasingly unable to conduct daily activities, which has a substantial personal and socioeconomic impact. Fairly recent prior studies showed that patients with RA stop working 20 years earlier than age-matched controls. Factors related to sociodemographic, clinical, care and disease profiles might affect the loss of work capacity. The purpose of this study was to assess the factors associated with the prevalence of working patients with rheumatoid arthritis in the municipality of Blumenau.
Methods
A cross-sectional, population-based study was conducted between July 2014 and January 2015, with 296 individuals aged 20 years or older, male and female, living in Blumenau, Santa Catarina state, Brazil, and diagnosed with rheumatoid arthritis according to the 1987 American College of Rheumatology criteria. The prevalence of working patients with RA was assessed by employment status self-reporting during the interview. The chi-squared test, Wald test and Poisson regression analysis were used to test the possible associations between the independent variables and outcome.
Results
The prevalence of working patients with rheumatoid arthritis was 44.3%. Patients aged 20 to 59 years had a 90% higher prevalence of outcome than subjects aged 60 years or older. The prevalence of working patients was 132% and 73% higher among individuals with low income and high functional disability, measured using the Health Assessment Questionnaire (HAQ), respectively.
Conclusion
The prevalence of working RA patients was highest among adult patients with low income and high functional disability. The first variable is directly related to the individual characteristic, the second reflects the socioeconomic context of the patient, and the third reflects the degree of disability caused by the disease, which may be modifiable by health professionals
Hematobin is a novel immunomodulatory protein from the saliva of the horn fly Haematobia irritans that inhibits the inflammatory response in murine macrophages
No full textAbstract
Background
The horn fly Haematobia irritans is a blood-sucking ectoparasite responsible for substantial economic loss of livestock. Like other hematophagous arthropods species, the successful blood-feeding of H. irritans is highly dependent on the modulation of the host’s hemostasis and immune system. Here, we evaluated the biological activity of hematobin (HTB), a protein recently identified in the H. irritans saliva, on macrophage biology. The goal was to understand the putative interactions between the components of H. irritans saliva and the early host immune responses.
Results
Thioglycolate-elicited peritoneal macrophages from BALB/c mice were stimulated by lipopolysaccharide (LPS) plus interferon-γ (IFN-γ) in the presence or absence of recombinant HTB. The presence of the salivary protein in the cultures inhibited nitric oxide production and decreased the inducible nitric oxide synthase (iNOS) expression induced by LPS plus IFN-γ. The tumor necrosis factor-α (TNF-α) and interleukin-12p40 (IL-12p40) levels were also reduced in the macrophages pre-incubated with HTB; these findings correlated to the decreased NF-κB expression. The biological activities described here were not associated with changes in annexin V binding to macrophages suggesting that HTB does not induce cell death. In addition, the activity of HTB seems to be specific to macrophages because no changes were observed in lymphocyte proliferation or cytokine production.
Conclusions
We describe here the first bioactive salivary protein of H. irritans. We characterized its ability to modulate macrophage inflammatory response, and the results can help explain how horn flies modulate the host immune system to feed on blood
Bat rabies surveillance and risk factors for rabies spillover in an urban area of Southern Brazil
No full textAbstract\ud
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Background\ud
Bat rabies surveillance data and risk factors for rabies spillover without human cases have been evaluated in Curitiba, the ninth biggest city in Brazil, during a 6-year period (2010–2015). A retrospective analysis of bat complaints, bat species identification and rabies testing of bats, dogs and cats has been performed using methodologies of seasonal decomposition, spatial distribution and kernel density analysis.\ud
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Results\ud
Overall, a total of 1003 requests for bat removal have been attended to, and 806 bats were collected in 606 city locations. Bat species were identified among 13 genera of three families, with a higher frequency of Nyctinomops in the central-northern region and Molossidae scattered throughout city limits. Out of the bats captured alive, 419/806 (52.0%) healthy bats were released due to absence of human or animal contacts. The remaining 387/806 (48.0%) bats were sent for euthanasia and rabies testing, which resulted in 9/387 (2.32%) positives. Linear regression has shown an increase on sample numbers tested over time (regression: y = 2.02 + 0.17×; p < 0.001 and r2 = 0.29), as well as significant seasonal variation, which increases in January and decreases in May, June and July. The Kernel density analysis showed the center-northern city area to be statistically important, and the southern region had no tested samples within the period. In addition, a total of 4769 random and suspicious samples were sent for rabies diagnosis including those from dogs, cats, bats and others from 2007 to 2015. While all 2676 dog brains tested negative, only 1/1136 (0.088%) cat brains tested positive for rabies.\ud
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Conclusion\ud
Only non-hematophagous bats were collected during the study, and the highest frequency of collections occurred in the center-northern region of the city. Rabies spillover from bats to cats may be more likely due to the registered exposure associated with cats’ innate hunting habits, predisposing them to even closer contact with potentially infected bats. Although associated with a very low frequency of rabies, cats should always be included in rabies surveillance and vaccination programs.The authors are grateful to the Municipal Secretary of Health of Curitiba,\ud
ZCC, for providing the data archive, to the University of State of Sao Paulo\ud
(UNESP - Botucatu) and University of Sao Paulo (USP) for financial and\ud
technical support, and for all work by ZCC that directly or indirectly\ud
contributed for this study
Smoking-induced aggravation of experimental arthritis is dependent of aryl hydrocarbon receptor activation in Th17 cells
No full textAbstract\ud
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Background\ud
Epidemiologic studies have highlighted the association of environmental factors with the development and progression of autoimmune and chronic inflammatory diseases. Among the environmental factors, smoking has been associated with increased susceptibility and poor prognosis in rheumatoid arthritis (RA). However, the immune and molecular mechanism of smoking-induced arthritis aggravation remains unclear. The transcription factor aryl hydrocarbon receptor (AHR) regulates the generation of Th17 cells, CD4 T cells linked the development of autoimmune diseases. AHR is activated by organic compounds including polycyclic aromatic hydrocarbons (PAHs), which are environmental pollutants that are also present in cigarette smoke. In this study, we investigated the role of AHR activation in the aggravation of experiment arthritis induced by exposure to cigarette smoke.\ud
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Methods\ud
Mice were exposed to cigarette smoke during the developmental phase of antigen-induced arthritis and collagen-induced arthritis to evaluate the effects of smoking on disease development. Aggravation of articular inflammation was assessed by measuring neutrophil migration to the joints, increase in articular hyperalgesia and changes in the frequencies of Th17 cells. In vitro studies were performed to evaluate the direct effects of cigarette smoke and PAH on Th17 differentiation. We also used mice genetically deficient for AHR (Ahr KO) and IL-17Ra (Il17ra KO) to determine the in vivo mechanism of smoking-induced arthritis aggravation.\ud
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Results\ud
We found that smoking induces arthritis aggravation and increase in the frequencies of Th17 cells. The absence of IL-17 signaling (Il17ra KO) conferred protection to smoking-induced arthritis aggravation. Moreover, in vitro experiments showed that cigarette smoke can directly increase Th17 differentiation of T cells by inducing AHR activation. Indeed, Ahr KO mice were protected from cigarette smoke-induced arthritis aggravation and did not display increase in TH17 frequencies, suggesting that AHR activation is an important mechanism for cigarette smoke effects on arthritis. Finally, we demonstrate that PAHs are also able to induce arthritis aggravation.\ud
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Conclusions\ud
Our data demonstrate that the disease-exacerbating effects of cigarette smoking are AHR dependent and environmental pollutants with AHR agonist activity can induce arthritis aggravation by directly enhancing Th17 cell development.The research leading to these results received funding from the European\ud
Union Seventh Framework Programme [FP7–2007-2013] under grant\ud
agreement number HEALTH-F4–2011-281608 (TIMER), from the São Paulo Research\ud
Foundation (FAPESP) under grant agreements number 2011/19670–0\ud
(Projeto Temático) and 2013/08216–2 (Center for Research in Inflammatory\ud
Fig. 4 AHR activation in T cells induces arthritis aggravation in an IL-17RA-dependent manner. a AIA was induced in WT or Il17ra−/− mice and\ud
treated with vehicle or FICZ (90 μg/kg) during immunization. Neutrophil in the joints and hyperalgesia were determined 7 h after challenge.\ud
Mean ± SEM, n = 5/group, *\ud
P < 0.05; n.s, not significant, one-way ANOVA followed by Bonferroni post hoc test. b AIA was induced in WT mice\ud
treated with anti-IL-17 (α-IL-17) or the isotype control antibody at the same time as the mice were treated with vehicle or FICZ. Neutrophil in the\ud
joints and hyperalgesia were determined 7 h after challenge. Mean ± SEM, n = 5/group, *\ud
P < 0.05; n.s, not significant, one-way ANOVA followed by\ud
Bonferroni post hoc test. c AIA was induced in WT or Ahr−/− mice. The mice were treated or not with FICZ. One group of Ahr−/− mice (CD4+\ud
WT\ud
cells → Ahr−/−\ud
) received CD4+ T cells from WT mice 1 day before the first immunization. Data are mean ± SEM, n = 5/group, *\ud
P < 0.05, **P < 0.01, ***P < 0.001, one-way ANOVA followed by Bonferroni post hoc test. All data are representative of two independent experiments. DLN draining\ud
lymph nodes, FICZ 6-formylindolo[3,2-b]carbazole, IL-17 interleukin 17\ud
Talbot et al. Arthritis Research & Therapy (2018) 20:119 Page 9 of 11\ud
Disease) and from the University of São Paulo NAP-DIN under grant agreement\ud
number 11.1.21625.01.0
Pulmonary arterial hypertension in Latin America: epidemiological data from local studies
No full textAbstract\ud
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Background\ud
Pulmonary arterial hypertension is a rare, progressive disease with poor prognosis. However, there is limited information available on the characteristics of PAH patients outside of North America and Europe. This is particularly important as researchers have described that there are potential geographical and regional differences which are vital to consider in the design of clinical trials as well as PAH treatment. The aim of this study was to describe the epidemiology of PAH (PH group 1) in Latin America.\ud
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Methods\ud
A search of electronic databases for studies published in English, Spanish or Portuguese was conducted specifying publication dates from the 1st of January 1987 until 10th October 2016. Two authors independently assessed papers for inclusion and extracted data. A narrative synthesis of the findings was conducted.\ud
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Results\ud
The search revealed 22 conference abstracts and articles, and on application of the inclusion criteria, six conference abstracts and articles were included in the final review. Studies/registries were based in Argentina, Brazil and Chile. In contrast to the available literature from developed countries, in Latin America, most patients were diagnosed at younger age; nevertheless, the higher prevalence of idiopathic PAH (IPAH) and the advanced stage of the disease at diagnosis were comparable to the existing literature, as the long term survival, despite the lower availability of targeted therapies.\ud
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Conclusion\ud
This study highlights the regional characteristics in the epidemiology of group 1 PH. The recognition of these differences should be considered when developing clinical guidelines and extrapolating diagnostic and treatment algorithms. Equitable access to health care and therapies are also issues that need to be addressed in Latin America. Information coming from a large prospective registry representing the different populations in Latin America is of critical importance to increase disease awareness in the region and improve diagnosis and management.This study (GSK study number LS2579) was funded and supported by\ud
GlaxoSmithKline (GSK). Dr. Rogerio Souza received no funding from GSK to\ud
work on this study. RANDOM Foundation (Colombia) provided medical\ud
writing services funded by GSK