Circulating Levels of Apelin, GDF-15 and Sarcopenia: Lack of Association in the MAPT Study.

OBJECTIVES: Apelin and GDF-15 have been proposed as biomarkers of age-related sarcopenia but evidence in human models is scarce. This study aimed to explore the associations between blood apelin and GDF-15 with sarcopenia incidence and the evolution of sarcopenia components over two years in older adults >70 years. DESIGN: Secondary longitudinal analysis of the Multidomain Alzheimer Preventive Trial. PARTICIPANTS: Older adults (>70 years) attending primary care centers in France and Monaco. SETTING: Community. MEASUREMENTS: Serum Apelin (pg/mL) and plasma GDF-15 (pg/mL) were measured. Outcomes included sarcopenia defined by the European Working Group on Sarcopenia in Older People (EWGSOP) and its determinants (appendicular lean mass [ALM] evaluated through a Dual-energy X-ray Absorptiometry (DXA) scan, handgrip strength (HGS) and the 4-meter gait speed) measured over 2 years. Linear mixed models and logistic regression were used to explore the longitudinal associations. RESULTS: We included 168 subjects from MAPT (median age=76y, IQR=73-79; 78% women). Serum apelin was not significantly associated with sarcopenia incidence (OR=1.001;95%CI=1.000,1.001;p-value>0.05 in full-adjusted models) nor with ALM (ß=-5.8E-05;95%CI=-1.0E-04,2.12E-04;p>0.05), HGS (ß=-1.1E-04;95%CI=-5.0E-04,2.8E-04;p>0.05), and GS (ß=-5.1E-06;95%CI=-1.0E-05,2.0E-05;p>0.05) in fully adjusted models. Similarly, plasma GDF-15 was not associated with both the incidence of sarcopenia (OR=1.001,95%CI=1.000,1.002,p>0.05) and the evolution of its determinants ([ALM, ß=2.1E-05;95%CI=-2.6E-04,3.03E-04;p>0.05], HGS [ß=-5.9E-04;95%CI=-1.26E-03,8.1E-05; p>0.05] nor GS [ß=-2.6E-06;95%CI=-3.0E-05, 2.3E-05;p>0.05]) in fully adjusted models. CONCLUSIONS: Blood apelin and GDF-15 were not associated with sarcopenia incidence or with the evolution of sarcopenia components over a 2-year follow-up in community-dwelling older adults. Well-powered longitudinal studies are needed to confirm or refute our findings.

Changes in mitochondrial morphology modulate LPS-induced loss of calcium homeostasis in BV-2 microglial cells.

Microglial activation involves both fragmentation of the mitochondrial network and changes in cellular Ca2+ homeostasis, but possible modifications in mitochondrial calcium uptake have never been described in this context. Here we report that activated microglial BV-2 cells have impaired mitochondrial calcium uptake, including lower calcium retention capacity and calcium uptake rates. These changes were not dependent on altered expression of the mitochondrial calcium uniporter. Respiratory capacity and the inner membrane potential, key determinants of mitochondrial calcium uptake, are both decreased in activated microglial BV-2 cells. Modified mitochondrial calcium uptake correlates with impaired cellular calcium signaling, including reduced ER calcium stores, and decreased replenishment by store operated calcium entry (SOCE). Induction of mitochondrial fragmentation through Mfn2 knockdown in control cells mimicked this effect, while inhibiting LPS-induced mitochondrial fragmentation by a dominant negative form of Drp1 prevented it. Overall, our results show that mitochondrial fragmentation induced by LPS promotes altered Ca2+ homeostasis in microglial cells, a new aspect of microglial activation that could be a key feature in the inflammatory role of these cells.

Isotropic finishing of austempered iron casting cylindrical parts by roller burnishing.

Roller burnishing technique to achieve isotropic surface topography on cylindrical components made of austempered ductile iron (ADI) casting is presented in this paper. In the last years, ADI casting components are used in many mechanical applications, due to their enhanced mechanical properties. ADI castings are difficult-to-cut materials; therefore, advanced techniques to improve manufacturing productivity are necessary and under research. On the other hand, spiral roughness pattern produced by turning operation is a common source of unconformities in several applications. Turning produces a defined kinematic pattern, similar to a thread. This work presents a theoretical and experimental validation using different burnishing conditions. Roughness and surface topography and surface integrity were checked. Results show that the technique greatly improves surface roughness, and eliminates the kinematic-driven roughness pattern of turning, leading to a more isotropic finishing. A comparison between roller burnishing and ball burnishing is also presented in this paper.

Proxy indicators to estimate the appropriateness of medications prescribed by paediatricians in infectious diseases: a cross-sectional observational study based on reimbursement data.

BACKGROUND: We previously developed proxy indicators (PIs) that can be used to estimate the appropriateness of medications used for infectious diseases (in particular antibiotics) in primary care, based on routine reimbursement data that do not include clinical indications. OBJECTIVES: To: (i) select the PIs that are relevant for children and estimate current appropriateness of medications used for infectious diseases by French paediatricians and its variability while using these PIs; (ii) assess the clinimetric properties of these PIs using a large regional reimbursement database; and (iii) compare performance scores for each PI between paediatricians and GPs in the paediatric population. METHODS: For all individuals living in north-eastern France, a cross-sectional observational study was performed analysing National Health Insurance data (available at prescriber and patient levels) regarding antibiotics prescribed by their paediatricians in 2017. We measured performance scores of the PIs, and we tested their clinimetric properties, i.e. measurability, applicability and room for improvement. RESULTS: We included 116 paediatricians who prescribed a total of 44 146 antibiotic treatments in 2017. For all four selected PIs (seasonal variation of total antibiotic use, amoxicillin/second-line antibiotics ratio, co-prescription of anti-inflammatory drugs and antibiotics), we found large variations between paediatricians. Regarding clinimetric properties, all PIs were measurable and applicable, and showed high improvement potential. Performance scores did not differ between these 116 paediatricians and 3087 GPs. CONCLUSIONS: This set of four proxy indicators might be used to estimate appropriateness of prescribing in children in an automated way within antibiotic stewardship programmes.

Spinal Myoclonus: Is It An Anesthetic Mystery? / Mioclonía espinal: ¿un misterio anestésico? ¡Se acepta el reto!

The goal is to present a descriptive study related an unprecedent case of spinal myoclonus (SM) following subarachnoid anesthesia (SA). SM are sudden, brief, involuntary non-generalized spasms that can be an adverse effect of drug administration via neuraxial routes. Female, 67y, ASA II, proposed for hip replacement surgery, with normal preoperative exams. 7min after SA with 10mg of bupivacaine 0,5%, no motor blockade observed, and patient complained of unbearable pain in legs and perineum and bilateral, asymmetrical and arrhythmic myoclonic movements in the lower limbs. The latter solved after 48h of general anesthesia and rocuronium perfusion, amongst other therapeutics. Accordingly, intrathecal bupivacaine appears to be the SM most likely cause, regarding the absence of neurologic and electrolyte disorders, spinal cord direct trauma, drug exchange and normal perioperative examination, imaging and laboratory testing.It is mandatory to always take the patients' anaesthetic histories and recognize, treat and report rare anaesthetic complications.

Fermentation profile and microbial population in soybean silages with inoculant and powdered molasses / Perfil fermentativo e população microbiana em silagens de soja com inoculante e melaço em pó

Fermentation profile and microbial population were assessed in soybean silages without any additive (control), with inoculant (I), with I + powdered molasses (I+M), and with powdered molasses only (M). Soybean plants were harvested at the R6 stage and ensiled in 2kg-capacity laboratory silos. The additives were added to the natural matter base of silages. The assessed fermentation periods were 1, 3, 7, 14, 28, and 56 days. A 4 × 6 factorial arrangement (4 additives × 6 fermentation periods) in a completely randomized design with 3 replicates was used. Lactic, acetic, and butyric acids concentrations were influenced by additives and periods (P< 0.05). It was observed higher lactic acid values to control silages, on the 56th day. Lower average values of acetic and butyric acids were observed to I+M and M silages. It was observed quadratic effect to pH values with a reduction estimated of 0.5504, 0.5358, 0.6312 and 0.6680 units to pH values to control, I, I+M, and M silages in the first 10 days. A maximum lactic acid bacteria population was observed at the 28th day of fermentation in silages with inoculant. The inoculant and powdered molasses improve the fermentation profile of soybean silages.(AU)

Avaliou-se o perfil fermentativo e a composição microbiana de silagens de soja sem aditivos (controle), com inoculante (I), com I + melaço em pó (I+M) e com melaço em pó (M). As plantas de soja foram colhidas no estádio R6 e ensiladas em silos laboratoriais com capacidade de 2kg. Os aditivos foram adicionados às silagens na matéria natural. Os períodos de fermentação avaliados foram: 1, 3, 7, 14, 28 e 56 dias. Utilizou-se um esquema fatorial 4 × 6 (quatro aditivos × seis períodos), em um delineamento completamente ao acaso, com três repetições. Observou-se efeito de aditivos e de período (P<0,05) sobre os ácidos láctico, acético e butírico. Foi constatado maior teor de ácido láctico na silagem controle aos 56 dias. Para os teores dos ácidos acético e butírico, registraram-se menores valores médios nas silagens I+M e M. Observou-se efeito quadrático para o pH das silagens, com estimativa de redução de pH nos primeiros 10 dias igual a 0,5504; 0,5358; 0,6312 e 0,6680 nas silagens controle, I, I+M e M, respectivamente. A máxima população de bactéria ácido-láctica foi observada aos 28 dias nas silagens inoculadas. O inoculante e o melaço em pó melhoram o perfil fermentativo de silagens de soja.(AU)

[Intracranial hemorrhage and oral anticoagulants of patients treated between 2011 and 2013 at the Nancy Regional University Hospital]. / Hémorragies intracrâniennes et anticoagulantes orales de patients traités entre 2011 à 2013. Étude descriptive au CHRU de Nancy.

OBJECTIVE: To perform a descriptive analysis of intracranial hemorrhages of patients treated with an antivitamin K (fluindione, acenocoumarol or warfarin) or a direct oral anticoagulant (dabigatran, rivaroxaban or apixaban) at the Nancy Regional University Hospital. MATERIAL AND METHOD: The study period was from January 2011 to December 2013 and the computerized data (Programme de Médicalisation des Systèmes d'Information) of our hospital was accessed to identify the patients. Clinical data were obtained from the patients' files. Regional healthcare system was queried for reimbursement data. RESULTS: Among the 157 identified cases of intracranial hemorrhage, 153 were related to antivitamin K, primarily fluindione (n=127), and only 4 to a direct oral anticoagulant (3 dabigatran and 1 rivaroxaban). During the same period, regional data indicated that 65,345 patients had had at least one reimbursement of antivitamin K and 20,983 patients one reimbursement of an oral direct anticoagulant. In our series, the most frequent intracranial hemorrhages were subdural hematoma (chronic in 65 cases, acute in 50 cases) and intraparenchymal hemorrhage (20 cases). The global mortality rate was 20.2% but varied with the site of hemorrhage. In multivariate analysis, the two risk factors of fatal outcome were coma on admission (OR 6.2; 95%CI: 2.6-15.0) and a history of previous intracranial hemorrhage (OR 13,4; 95% CI: 1,6-114,9). CONCLUSION: During the 2011-2013 period, antivitamin K, especially fluindione, was the most frequently involved anticoagulants in intracranial hemorrhages with hospitalization at our Regional University Hospital. Coma on admission and a history of previous intracranial hemorrhage were the two main risk factors for fatal outcome.

The capacity of silage inoculant bacteria to bind aflatoxin B1 in vitro and in artificially contaminated corn silage.

The objectives were to examine the aflatoxin B1 (AFB1)-binding capacity of silage bacteria and factors affecting the responses. Experiments 1 and 2 examined the effects of bacterial strain and population on the AFB1-binding capacity of 10 bacteria. When applied at 106 cfu/mL to an in vitro medium, only Lactobacillus plantarum PT5B bound the AFB1 and the binding capacity was low (4%). When applied at 109 cfu/mL, all 10 bacteria bound AFB1, but L. plantarum R2014 (Lp) and EQ12, Lactobacillus buchneri R1102 (Lb), and Pediococcus acidilactici R2142 and EQ01 (Pa) had the greatest capacity (23.9 to 33%). Experiment 3 examined the AFB1-binding capacity of viable and nonviable (HCl-treated) forms of Lp, Lb, and Pa at different pH. Nonviable Lb and Lp, but not Pa, increased AFB1 binding. Binding of AFB1 was greatest at pH 2.5 and least at pH 8. As the nonviable Lb and Lp that bound AFB1 in experiment 3 would not be effective silage inoculants, experiment 4 examined effects of benign versus severe treatments (85 vs. 100°C; pH 2.5 vs. <1) on the viability of Lp, Lb, and Pa. The population of bacteria was reduced from 9 to 4 log cfu/mL by treatment with HCl at pH 2.5 and to 2 log cfu/mL by 85 or 100°C, whereas acidification at pH <1 eliminated the bacteria. Experiment 5 determined the effect of the ensiling duration and benign treatment methods [37 (viable cells) or 85°C (heated cells) or acidification with HCl at pH 2.5 (acid-treated cells)] on binding of AFB1 and silage quality during the fermentation of corn forage. Corn forage was ensiled after treatment with only deionized water (control), AFB1 (30 µg/kg of fresh forage), or a mixture of AFB1 and 109 cfu/g of each of the treated bacteria. Adding AFB1 alone to corn forage reduced the pH decline during the first 3 d of ensiling and increased or tended to increase butyric acid concentration and final pH after ensiling for 21 d. Bacterial inoculation inhibited these negative effects. The fermentation profile of silage treated with Lb and Pa did not differ from those of the control silage. In all silages treated with the toxin, the AFB1 concentration decreased linearly (from 30 to ≤0.35 µg/kg) within 3 d of ensiling. Certain silage bacteria can bind AFB1 but the efficacy depends on several factors.

Phylogeny and taxonomy of the scab and spot anthracnose fungus Elsinoë (Myriangiales, Dothideomycetes).

Species of Elsinoë are phytopathogens causing scab and spot anthracnose on many plants, including some economically important crops such as avocado, citrus, grapevines, and ornamentals such as poinsettias, field crops and woody hosts. Disease symptoms are often easily recognisable, and referred to as signature-bearing diseases, for the cork-like appearance of older infected tissues with scab-like appearance. In some Elsinoë-host associations the resulting symptoms are better described as spot anthracnose. Additionally the infected plants may also show mild to severe distortions of infected organs. Isolation of Elsinoë in pure culture can be very challenging and examination of specimens collected in the field is often frustrating because of the lack of fertile structures. Current criteria for species recognition and host specificity in Elsinoë are unclear due to overlapping morphological characteristics, and the lack of molecular and pathogenicity data. In the present study we revised the taxonomy of Elsinoë based on DNA sequence and morphological data derived from 119 isolates, representing 67 host genera from 17 countries, including 64 ex-type cultures. Combined analyses of ITS, LSU, rpb2 and TEF1-α DNA sequence data were used to reconstruct the backbone phylogeny of the genus Elsinoë. Based on the single nomenclature for fungi, 26 new combinations are proposed in Elsinoë for species that were originally described in Sphaceloma. A total of 13 species are epitypified with notes on their taxonomy and phylogeny. A further eight new species are introduced, leading to a total of 75 Elsinoë species supported by molecular data in the present study. For the most part species of Elsinoë appear to be host specific, although the majority of the species treated are known only from a few isolates, and further collections and pathogenicity studies will be required to reconfirm this conclusion.

Autoimmune hemolytic anemia in systemic lupus erythematosus at diagnosis: differences between pediatric and adult patients.

Objective To determine the overall prevalence of autoimmune hemolytic anemia (AIHA), and to compare clinical and laboratory features in a large population of children and adult lupus patients at diagnosis. Methods This retrospective study evaluated the medical charts of 336 childhood-onset systemic lupus erythematosus (cSLE) and 1830 adult SLE (aSLE) patients followed in the same tertiary hospital. Demographic data, clinical features and disease activity were recorded. AIHA was defined according to the presence of anemia (hemoglobin <10 g/dL) and evidence of hemolysis (reticulocytosis and positive direct antiglobulin test (DAT)/Coombs test) at SLE diagnosis. Evans syndrome (ES) was defined by the combination of immune thrombocytopenia (platelet count <100,000/mm3) and AIHA. Results The frequency of AIHA at diagnosis was significantly higher in cSLE patients compared to aSLE (49/336 (14%) vs 49/1830 (3%), p = 0.0001), with similar frequency of ES (3/336 (0.9%) vs 10/1830 (0.5%), p = 0.438). The median of hemoglobin levels was reduced in cSLE vs aSLE patients (8.3 (2.2-10) vs 9.5 (6.6-10) g/dL, p = 0.002) with a higher frequency of multiple hemorrhagic manifestations (41% vs 7%, p = 0.041) and erythrocyte transfusion due to bleeding (24% vs 5%, p = 0.025). cSLE patients also had more often constitutional involvement (84% vs 31%, p < 0.001), fever (65% vs 26%, p < 0.001), weight loss > 2 kg (39% vs 6%, p < 0.001), reticuloendothelial manifestations (48% vs 8%, p < 0.001), hepatomegaly (25% vs 2%, p < 0.001) and splenomegaly (21% vs 2%, p = 0.004). Other major organ involvements were common but with similar frequencies in cSLE and aSLE ( p > 0.05). Median systemic lupus erythematosus disease activity index 2000 (SLEDAI-2 K) was comparable in cSLE and aSLE (p = 0.161). Conclusions We identified that AIHA was not a common condition in cSLE and aSLE, with distinct features characterized by a higher prevalence/severity in children and concomitant constitutional symptoms in the majority of them.

Blood levels of pro-inflammatory and anti-inflammatory cytokines during an oral glucose tolerance test in patients with symptoms suggesting reactive hypoglycemia.

We evaluated the impact of postprandial glycemia on blood levels of pro-inflammatory and anti-inflammatory cytokines during an oral glucose tolerance test in non-diabetic patients with symptoms suggesting reactive hypoglycemia. Eleven patients with clinical symptoms suggesting reactive hypoglycemia received an oral glucose solution (75 g) Blood was collected at 0 (baseline), 30, 60, 120 and 180 min after glucose ingestion and the plasma concentrations of interferon-α (IFN-α), interferon-γ (IFN-γ), interleukin-1 receptor antagonist (IL-1RA), interleukin 2 (IL-2), interleukin-2 receptor (IL-2R), interleukin 4 (IL-4), interleukin 6 (IL-6), interleukin 8 (IL-8), interleukin 10 (IL-10), interleukin-12 (IL-12), interleukin 13 (IL-13), interleukin 15 (IL-15), interleukin 17 (IL-17), IFN-γ inducible protein 10 (IP-10), monocyte chemotactic protein 1 (MCP1), monokine induced by IFN-γ (MIG), macrophage inflammatory protein-1α (MIP-1α), interleukin-1ß (IL-1ß), colony stimulating factor (G-CSF), granulocyte-macrophage CSF (GM-CSF), basic fibroblast growth factor (FGF-basic), eotaxin, tumor necrosis factor α (TNFα), epidermal growth factor (EGF), hepatocyte growth factor (HGF), vascular endothelial growth factor (VEGF), macrophage inflammatory protein-1α (MIP-1α), and 1ß (MIP-1ß) were evaluated. Overall, glycemic levels increased, reached its maximum at 30 min (phase 1), returned to baseline levels at 120 min (phase 2), followed by a mild hypoglycemia at 180 min (phase 3). During phase 1, cytokine blood levels were maintained. However, we observed a synchronous fall (P<0.05) in the concentrations of pro-inflammatory (IL-15, IL-17, MCP-1) and anti-inflammatory cytokines (FGF-basic, IL-13, IL-1RA) during phase 2. Furthermore, a simultaneous rise (P<0.05) of pro-inflammatory (IL-2, IL-5, IL-17) and anti-inflammatory cytokines (IL-4, IL-1RA, IL-2R, IL-13, FGF-basic) occurred during phase 3. Thus, mild acute hypoglycemia but not a physiological increase of glycemia was associated with increased blood levels of anti-inflammatory and pro-inflammatory cytokines.

Effects of lactic acid bacteria with bacteriocinogenic potential on the fermentation profile and chemical composition of alfalfa silage in tropical conditions.

The fermentation profile, chemical composition, and microbial populations of alfalfa silages treated with microbial inoculants (MI) at different fermentation periods (T) were evaluated in tropical conditions. A 4×6 factorial arrangement was used in a randomized design with 3 replicates. Fresh alfalfa was treated with (1) no treatment (CTRL), (2) commercial inoculant (CIN), (3) Pediococcus acidilactici (strain 10.6, S1), and (4) Pediococcus pentosaceus (strain 6.16, S2). An inoculant application rate of 10(6) cfu/g of fresh forage was used. The fermentation periods were 1, 3, 7, 14, 28, and 56 d. Alfalfa was harvested 82 d after sowing at the early flowering stage, chopped into 1.5-cm particle size, and ensiled in 25 × 35 cm vacuum-sealed plastic bags. The numbers of lactic acid bacteria, enterobacteria, mold, and yeast in alfalfa before ensiling were 5.42, 5.58, 4.82, and 4.8 log cfu/g, respectively. Silage chemical composition was evaluated only at 56 d. All parameters were affected by the interaction MI × T, except the concentrations of lactic and propionic acids. Alfalfa silage treated with S1 or S2 had lower pH values than CTRL from the first day until 28 d. However, the inoculants resulted in similar pH after 56 d, and these values were lower than the CTRL. The highest concentration of lactic acid was observed in the silage treated with S1 and S2 at 7 and 14 d of ensiling. The concentration of acetic acid was lower in the silages treated with S1 and S2 than the CTRL and CIN at 3 and 28 d of fermentation. There was no effect of MI or MI × T interaction on the microbial populations. However, the number of enterobacteria decreased over the fermentation period until 14 d and increased slightly after this time point. The chemical composition of alfalfa silage was not affected by MI at 56 d of ensiling. The strain P. pentosaceus 6.16 was the most efficient in dominating the fermentation process by decreasing the pH more quickly and increasing the concentration of lactic acid, suggesting its potential use as a silage inoculant.

High genetic variability in endophytic fungi from the genus Diaporthe isolated from common bean (Phaseolus vulgaris L.) in Brazil.

AIMS: The goals of the present study were to identify, to analyse the phylogenetic relations and to evaluate the genetic variability in Diaporthe endophytic isolates from common bean. METHODS AND RESULTS: Diaporthe sp., D. infecunda and D. phaseolorum strains were identified using multilocus phylogeny (rDNA ITS region; EF1-α, ß-tubulin, and calmodulin genes). IRAP (Inter-Retrotransposon Amplified Polymorphism) and REMAP (Retrotransposon-Microsatellite Amplified Polymorphism) molecular markers reveal the existence of high genetic variability, especially among D. infecunda isolates. CONCLUSIONS: It was concluded that the multilocus phylogenetic approach was more effective than individual analysis of ITS sequences, in identifying the isolates to species level, and that IRAP and REMAP markers can be used for studying the genetic variability in the genus Diaporthe particularly at the intraspecific level. SIGNIFICANCE AND IMPACT OF THE STUDY: The combined use of molecular tools such as multilocus phylogenetic approach and molecular markers, as performed in this study, is the best way to distinguish endophytic strains of Diaporthe isolated from common bean (Phaseolus vulgaris L.).

Exploring fungal mega-diversity: Pseudocercospora from Brazil.

Although the genus Pseudocercospora has a worldwide distribution, it is especially diverse in tropical and subtropical countries. Species of this genus are associated with a wide range of plant species, including several economically relevant hosts. Preliminary studies of cercosporoid fungi from Brazil allocated most taxa to Cercospora, but with the progressive refinement of the taxonomy of cercosporoid fungi, many species were relocated to or described in Pseudocercospora. Initially, species identification relied mostly on morphological features, and thus no cultures were preserved for later phylogenetic comparisons. In this study, a total of 27 Pseudocercospora spp. were collected, cultured, and subjected to a multigene analysis. Four genomic regions (LSU, ITS, tef1 and actA) were amplified and sequenced. A multigene Bayesian analysis was performed on the combined ITS, actA and tef1 sequence alignment. Our results based on DNA phylogeny, integrated with ecology, morphology and cultural characteristics revealed a rich diversity of Pseudocercospora species in Brazil. Twelve taxa were newly described, namely P. aeschynomenicola, P. diplusodonii, P. emmotunicola, P. manihotii, P. perae, P. planaltinensis, P. pothomorphes, P. sennae-multijugae, P. solani-pseudocapsicicola, P. vassobiae, P. wulffiae and P. xylopiae. Additionally, eight epitype specimens were designated, three species newly reported, and several new host records linked to known Pseudocercospora spp.

Blood levels of pro-inflammatory and anti-inflammatory cytokines during an oral glucose tolerance test in patients with symptoms suggesting reactive hypoglycemia

We evaluated the impact of postprandial glycemia on blood levels of pro-inflammatory and anti-inflammatory cytokines during an oral glucose tolerance test in non-diabetic patients with symptoms suggesting reactive hypoglycemia. Eleven patients with clinical symptoms suggesting reactive hypoglycemia received an oral glucose solution (75 g) Blood was collected at 0 (baseline), 30, 60, 120 and 180 min after glucose ingestion and the plasma concentrations of interferon-α (IFN-α), interferon-γ (IFN-γ), interleukin-1 receptor antagonist (IL-1RA), interleukin 2 (IL-2), interleukin-2 receptor (IL-2R), interleukin 4 (IL-4), interleukin 6 (IL-6), interleukin 8 (IL-8), interleukin 10 (IL-10), interleukin-12 (IL-12), interleukin 13 (IL-13), interleukin 15 (IL-15), interleukin 17 (IL-17), IFN-γ inducible protein 10 (IP-10), monocyte chemotactic protein 1 (MCP1), monokine induced by IFN-γ (MIG), macrophage inflammatory protein-1α (MIP-1α), interleukin-1β (IL-1β), colony stimulating factor (G-CSF), granulocyte-macrophage CSF (GM-CSF), basic fibroblast growth factor (FGF-basic), eotaxin, tumor necrosis factor α (TNFα), epidermal growth factor (EGF), hepatocyte growth factor (HGF), vascular endothelial growth factor (VEGF), macrophage inflammatory protein-1α (MIP-1α), and 1β (MIP-1β) were evaluated. Overall, glycemic levels increased, reached its maximum at 30 min (phase 1), returned to baseline levels at 120 min (phase 2), followed by a mild hypoglycemia at 180 min (phase 3). During phase 1, cytokine blood levels were maintained. However, we observed a synchronous fall (P<0.05) in the concentrations of pro-inflammatory (IL-15, IL-17, MCP-1) and anti-inflammatory cytokines (FGF-basic, IL-13, IL-1RA) during phase 2. Furthermore, a simultaneous rise (P<0.05) of pro-inflammatory (IL-2, IL-5, IL-17) and anti-inflammatory cytokines (IL-4, IL-1RA, IL-2R, IL-13, FGF-basic) occurred during phase 3. Thus, mild acute hypoglycemia but not a physiological increase of glycemia was associated with increased blood levels of anti-inflammatory and pro-inflammatory cytokines.

Diversity and potential impact of Calonectria species in Eucalyptus plantations in Brazil.

Species in the genus Calonectria (Hypocreales) represent an important group of plant pathogenic fungi that cause serious losses to plant crops in tropical and subtropical climates. Calonectria leaf blight is currently one of the main impediments to Eucalyptus cultivation in Brazil, and various species of Calonectria have been associated with this disease. Since most previous identifications were solely based on morphological characters, much of the published literature needs to be re-evaluated. The aim of this study was thus to identify and determine the phylogenetic relationships among species that occur in the Eucalyptus growing regions of Brazil by using partial sequences of the ß-tubulin, calmodulin, translation elongation factor 1-α and histone H3 gene regions. Based on extensive collections from soil and infected eucalypt leaf samples from plantations, phylogenetic inference revealed the Ca. pteridis complex to be the most common species complex present in Eucalyptus plantations in Brazil. By elucidating taxa in the Ca. pteridis, Ca. cylindrospora and Ca. candelabra species complexes, 20 novel Calonectria species were identified, and a new name in Calonectria provided for Cylindrocladium macrosporum as Ca. pseudopteridis.

First Report of Colletotrichum karstii Causing Anthracnose on Blueberry Leaves in Brazil.

Anthracnose or ripe rot of blueberry (Vaccinium spp.) is caused predominantly by Colletotrichum fioriniae, which belongs to the C. acutatum complex since it has cylindrical conidia with both ends acute (2). In May 2013, an isolate typical of the C. boninense complex (cylindrical conidia with both ends rounded) (1) was obtained from leaves of southern highbush blueberry seedlings in a nursery located in the municipality of Pelotas, Rio Grande do Sul, Brazil. The symptoms initially appear as circular, necrotic lesions 10 to 30 mm in diameter. At high humidity, lesions expand rapidly to cover the entire foliar surface, leading to severe defoliation of the seedlings. This disease occurred in 100% of the seedlings, causing serious losses in the nursery. A single-conidium culture was obtained on potato-dextrose-agar (PDA) medium followed by morphological and molecular characterization. This culture was deposited at the culture collection of the Universidade Federal de Viçosa, Brazil (Accession No. COAD 1741). Conidia were cylindrical, aseptate, hyaline, rounded at both ends, and 11 to 16 µm (µ = 13) long and 5 to 6 µm (µ = 5.5) wide (n = 100). For the molecular characterization, sequences of the internal transcribed spacer (ITS), glyceraldehyde-3-phosphate dehydrogenase (GAPDH) and ß-tubulin 2 (ßt) regions were obtained and deposited in GenBank (KM055653 to KM055655). A search of the Q-bank Fungi database using the ITS, ßt, and GAPDH sequences retrieved C. karstii with 100, 99, and 100% identities, respectively. Based on morphological and molecular data, the fungus was identified as C. karstii. To verify pathogenicity, 20 leaves from the upper branches of 1-m tall blueberry seedlings were inoculated with 6-mm-diameter plugs from a 7-day-old culture. PDA plugs were placed on the leaves of seedlings to serve as the control. Initially, seedlings were maintained at 25 ± 2°C in the dark. Thereafter, seedlings were covered with plastic bags and transferred to a greenhouse. Anthracnose symptoms on the leaves were observed at 5 days after inoculation. Seedlings from the control treatment remained symptomless. The fungus was re-isolated from the necrotic lesions, confirming Koch's postulates. C. karstii has a wide host range and in Brazil has been previously reported on Bombax aquaticum, Carica papaya, Eugenia uniflora, Malus domestica, and Mangifera indica (1,3,4). To the best of our knowledge, this is the first report of C. karstii causing anthracnose on the leaves of blueberry seedlings in Brazil or worldwide. Due to the high disease severity and intense defoliation of blueberry seedlings, this pathogen represents a new threat for nurseries. Therefore, control strategies should be investigated for this disease. References: (1) U. Damm et al. Stud. Mycol. 73:1, 2012. (2) U. Damm et al. Stud. Mycol. 73:37, 2012. (3) Lima et al. Plant Dis. 97:1248, 2014. (4) Velho et al. Plant Dis. 98:157, 2014.

Towards a phylogenetic reappraisal of Parmulariaceae and Asterinaceae (Dothideomycetes).

Members of the Asterinaceae and Parmulariaceae are obligate biotrophic fungi with a pantropical distribution that grow in direct association with living plant tissues and produce external ascomata and bitunicate asci. These fungi are poorly known, with limited information about their taxonomic position in the Dothideomycetes. Much of what is known is conjectural and based on observation of morphological characters. An assessment of the phylogenetic position of the Asterinaceae and Parmulariaceae is provided based on a phylogenetic analysis of the nrDNA operon (ITS) and the large subunit rDNA (LSU) sequence data obtained from fresh material of selected species collected in Brazil. Three key species were included and epitypified, namely Asterina melastomatis, which is the type species for the type genus of the Asterinaceae; Prillieuxina baccharidincola (Asterinaceae); and Parmularia styracis, which is the type species for the type genus of the Parmulariaceae. An LSU rDNA phylogenetic analysis was performed indicating the correct phylogenetic placement of the Asterinales within the Dothideomycetes. From this initial analysis it is clear that the Parmulariaceae as currently circumscribed is polyphyletic, and that the Asterinaceae and Parmulariaceae are related, which justifies the maintenance of the order Asterinales. Asterotexis cucurbitacearum is recognised as distinct from other Dothideomycetes and placed in the newly proposed family and order (Asterotexiaceae, Asterotexiales), while the higher order phylogeny of Inocyclus angularis remains unresolved. Additionally, Lembosia abaxialis is introduced as a novel species and the phylogenetic placement of the genera Batistinula and Prillieuxina is clarified.

Intake, digestibility, nitrogen efficiency, and animal performance of growing and finishing beef cattle fed warm-season legume (Stylosanthes capitata plus Stylosanthes macrocephala) silage replacing corn silage.

It was hypothesized that Stylosanthes cv. Campo Grande (ES) silage could be used as the single source of dietary forage for beef cattle and that performance on ES would be similar to corn silage (CS) at a 50:50 forage:concentrate. The objectives of this study were to evaluate intake, total and partial digestibility of nutrients, ruminal pH, ruminal ammonia, and productive performance in growing beef cattle fed diets with varying proportions of ES silage replacing CS. Treatments consisted of diets with ratios of 0:100, 25:75, 50:50, 75:25, and 100:0% ES:CS. Two experiments were conducted simultaneously. In the first experiment, 10 crossbred Holstein-Zebu bulls with an average initial weight of 272 ± 86 kg were used. The bulls were rumen and abomasums fistulated. An experimental design of two 5 × 5 Latin squares (Exp. 1) was used. The second experiment used 40 Nellore bulls with an average BW of 386 ± 30 kg in a completely randomized design (Exp. 2). Results showed a linear increase in CP intake (P < 0.05) in response to increased dietary ES. An increase in the proportion of ES in the diet had a negative linear effect on TDN. Apparent ruminal digestibility of CP increased linearly, and apparent intestinal digestibility of nonfibrous carbohydrates increased with the addition of ES to the diet (P < 0.05). Intestinal digestibility of DM exhibited a quadratic response (P < 0.05). Nitrogen balance, excretion of urinary urea, and plasma urea nitrogen did not respond to the inclusion of ES in the diet (P > 0.05). There was also no effect (P > 0.05) of ES inclusion on animal performance. Ruminal pH was not affected by an increased proportion of ES in the diet (P > 0.05), but ruminal pH was affected (P < 0.05) by the time of collection, for which a cubic model fit the data. There was an interaction (P < 0.05) between treatment and collection time for ruminal ammonia nitrogen concentration. It can be concluded that ES silage can be used as a source of roughage in the diet of beef cattle during the growing and finishing phases at a proportion of 50% of DM in the total diet. Therefore, ES silage is a promising alternative dietary ingredient and the use of this alternative source of silage will depend on availability and economic factors.