The aim of this study was to evaluate whether changing the interval from CIDR removal to timed artificial insemination (TAI) according to the diameter of the preovulatory follicle (POF) would improve ...pregnancy per AI in cows. In Study 1, a retrospective analysis of TAI experiments (n=96 cows) was performed to characterize the time of ovulation according to the diameter of the dominant follicle. It was observed that cows with a larger POF had ovulations earlier than cows with smaller POF, according to the equation: y=0.72x2−26.74x+264.54 (R2=0.63; P<0.001). In Study 2, lactating Nelore cows (n=412) were subjected to an EB-CIDR based TAI protocol. On the morning of Day 10 (time of TAI), cows were randomized into Control (n=209) and Block (n=203) groups; (1) Cows in the Control Group were TAI 48h after CIDR removal (08:00am on Day 10), and; (2) Cows in the block group were inseminated once at one of the following time points, according to the diameter of the POF on Day 10: B0 (POF≥15mm, TAI 0h after convetional TAI), B1 (POF 13–14.9mm, TAI 6h later), B2 (POF 10.1–12.9mm, TAI 24h later) and B3 (POF≤10mm, TAI 30h later). The cows of the Block Group had greater pregnancy rates per AI than the Control Group (129/203, 63.5% when compared with 102/209, 48.8%, respectively; P<0.01). In conclusion, results of the present study demonstrate that adjusting the timing of TAI according to the diameter of the POF can be an effective practice for improving fertility of cows in TAI protocols.
The aim of this study was to evaluate if prostaglandin F2α (PGF) can be used to induce ovulation in a GnRH-progesterone based protocol. In Experiment 1 crossbred dairy cows (n=32) were synchronized ...with a progesterone-GnRH based protocol for seven days, where the luteolytic dose of 150μg PGF was given 24h prior progesterone device removal (CIDR). On Day 8 cows were separated into two groups to receive: 1) 2mL of Saline (Control Group, n=15) or 2) 150μg of PGF (PGF Group, n=17). Ovulation rate was higher in the PGF than Control group (100% vs 53.3%, P=0.001, Odds ratio=30.88). The percentage of cows that ovulated synchronously tended to be higher in the PGF than Control group (P=0.1, Odds ratio=9.6). Experiment 2 was performed in a cross-over (3×3) design. Crossbred dairy cows (n=25) received a CIDR for seven days and GnRH on Day 0. Seven days later 150μg of PGF was given and the progesterone device was removed, and 24h later cows were distributed into three groups to receive: 1) 2mL of Saline (Control Group, n=25), 2) 150μg of PGF (PGF Group, n=25) or 3) 1mg of ECP (ECP Group, n=23). Diameter of ovulatory follicle was larger in the PGF and Control than ECP Group (P=0.002, Effect size>4.0). Synchronized ovulation rate (between 72 and 96h after CIDR removal) tended to be higher in PGF group in Control group (P=0.1, Odds ratio=0.35). Results suggest that PGF is equally efficient to ECP to induce synchronized ovulation in dairy cows subjected to progesterone-GnRH based protocols.
•The effect of prostaglandin F2α (PGF) as ovulation inducer in a GnRH-progesterone based protocol was evaluate.•PGF was compared with estradiol cypionate in a protocol to cattle.•PGF is equally efficient to ECP to induce concentrated ovulation in dairy cows.
Ricinus communis L. is of great economic importance due to the oil extracted from its seeds. Castor oil has been used for pharmaceutical and industrial applications, as a lubricant or coating agent, ...as a component of plastic products, as a fungicide or in the synthesis of biodiesel fuels. After oil extraction, a castor cake with a large amount of protein is obtained. However, this by-product cannot be used as animal feed due to the presence of toxic (ricin) and allergenic (2S albumin) proteins. Here, we propose two processes for detoxification and allergen inactivation of the castor cake. In addition, we establish a biological test to detect ricin and validate these detoxification processes. In this test, Vero cells were treated with ricin, and cell death was assessed by cell counting and measurement of lactate dehydrogenase activity. The limit of detection of the Vero cell assay was 10 ng/mL using a concentration of 1.6 x 10(5) cells/well. Solid-state fermentation (SSF) and treatment with calcium compounds were used as cake detoxification processes. For SSF, Aspergillus niger was grown using a castor cake as a substrate, and this cake was analyzed after 24, 48, 72, and 96 h of SSF. Ricin was eliminated after 24 h of SSF treatment. The cake was treated with 4 or 8% Ca(OH)2 or CaO, and both the toxicity and the allergenic properties were entirely abolished. A by-product free of toxicity and allergens was obtained.
Submarine canyons play an important role in the transfer of contaminated sediments from shelf areas to the deeper ocean. To evaluate the importance of submarine canyons adjacent to the Tagus and Sado ...estuaries (Portuguese Margin) as sediment pathway major and trace elements,
210Pb radionuclides, organic carbon, calcium carbonate and grain size were analyzed in fifteen short sediment cores from the Cascais and Lisboa–Setúbal submarine canyons and neighbor slopes at the Estremadura Spur and off Sines. Mass accumulation rates (MAR) vary between 0.02 and 0.40
g cm
−2 y
−1 with the highest values in the upper Cascais and Lisboa canyons. Mercury and Pb enrichment factors exceed 2 and show a continuously increasing trend since 1950
AD in the upper sections of the Cascais Canyon and in the Lisboa–Setúbal Canyon system, implying an anthropogenic source. Mercury and Pb anthropogenic inventories also present highest values in the canyon heads, which is consistent with its proximity to the Tagus and Sado river mouths. To a lesser extent, the Estremadura and Sines slopes and the deeper sections of the canyons also have slightly increasing Hg and Pb enrichment factor trends towards the present-day. For these areas, with a major influence of pelagic sedimentation, we infer atmospheric transport as a complementary source of contaminants. Therefore, our data suggest that although canyons are preferential conduits to transport sediments to the deep sea when compared to the slopes, Hg and Pb derived from human activities have reached both domains with different intensities down to a depth of 2000
mwd.
► Submarine Canyons are preferential conduits of contaminated sediments. ► Mercury and Pb present the highest anthropogenic contributions. ► Areas with pelagic sedimentation show increasing trend Hg and Pb towards the Present.
In chronic myeloid leukemia (CML) cell lines, brief exposure to pharmacologically relevant dasatinib concentrations results in apoptosis. In this study, we assess the impact of intensity and duration ...of Bcr-Abl kinase inhibition on primary CD34(+) progenitors of chronic phase CML patients. As CML cells exposed to dasatinib in vivo are in a cytokine-rich environment, we also assessed the effect of cytokines (six growth factors cocktail or granulocyte-macrophage colony-stimulating factor (CSF) or granulocyte-CSF) in combination with dasatinib. In the presence of cytokines, short-term intense Bcr-Abl kinase inhibition (>or=90% p-Crkl inhibition) with 100 nM dasatinib did not reduce CD34(+) colony-forming cells (CFCs). In contrast, without cytokines, short-term exposure to dasatinib reduced CML-CD34(+) CFCs by 70-80%. When cytokines were added immediately after short-term exposure to dasatinib, CML-CD34(+) cells remained viable, suggesting that oncogene dependence of these cells can be overcome by concomitant or subsequent exposure to cytokines. Additional inhibition of Janus tyrosine kinase (Jak) activity re-established the sensitivity of CML progenitors to intense Bcr-Abl kinase inhibition despite the presence of cytokines. These findings support the contention that therapeutic strategies combining intense Bcr-Abl kinase inhibition and blockade of cytokine signaling pathways can be effective for eradication of CML progenitors.
Type 2 diabetes mellitus (T2DM) consists of a set of metabolic and endocrine disorders which evolve into deficiency in insulin action and hyperglycemia. Physical exercise is considered the main ...intervention to prevent and control T2DM. Literature has suggested that circulating microRNAs (miRs) help to understand responses to physical activity among diabetic patients. Thus, the aim of this study was to analyze the acute effect of two interventions (strength and cardiovascular) on the total, whole blood circulating concentrations of miR-126, miR-146a and miR-155 in older adults with and without T2DM. A total of 23 male and female older adults (68.2±5.3 years) participated in the trial, 13 of whom presented with controlled T2DM and 10 were nondiabetics. They underwent both interventions separately, performed with intensity from 60% to 70% of reserve heart rate. Glucose and miRs levels were quantified and compared across groups with baseline titers as covariables. Diabetic patients showed more reduction in serum blood glucose than nondiabetics, with a great magnitude of reduction after the strength training intervention, which was paralleled by a positive change of the whole blood circulating levels of miR-146a, but not of the other miRs. Our report supports evidence that miR-146a levels in peripheral blood leukocytes are negatively associated with a state of insulin resistance, which is suggested as a novel marker to trace response to antidiabetic interventions.
This work presents some of the results from the project: “Effect of the environment on reinforcement durability” (DURACON) in its first two-years period, which investigates the influence of urban and ...marine meteorochemical parameters on the performance of reinforced concrete structures. The results presented in this investigation are from 21 marine test sites only (no urban environments are included), distributed among 11 countries (Argentina, Bolivia, Brazil, Chile, Colombia, Costa Rica, Mexico, Spain, Uruguay, Portugal and Venezuela). The environment was evaluated using ISO Standard 9223 and the concrete was characterized by measuring compressive strength, elastic modulus, total and effective porosity, chloride permeability according to ASTM standards, as well as the effective porosity and resistance to water absorption using the Fagerlund method. To that effect, concrete specimens (with and without reinforcement) were prepared for electrochemical and physical/mechanical/chemical tests using the existing materials in each participating country, following strict procedures which enabled the preparation of similar concrete samples. Two water/cement (w/c) ratios (0.45 and 0.65) were selected, where the concrete with 0.45 w/c ratio had to have a minimum cement content of 400
kg/m
3 and the one with 0.65 w/c ratio a compressive strength of 210
kg/cm
2. Type I Portland cement, siliceous sand, and crushed rock as coarse aggregates (13-mm maximum nominal size) were used. After a one-year exposure, the results of the corrosion potentiality and probability analysis of the reinforcement in the different test stations showed that, for marine atmospheres, the most aggressive environment to induce steel corrosion was at Portugal’s Cabo Raso station, and the least aggressive one was at Chile’s Valparaíso station. These results are comparable with the ones found using electrochemical measurements, after a two-year exposure.
Mast cell diseases are myeloproliferative neoplasms characterized by an abnormal proliferation and accumulation of mast cells in different tissues. The clinical presentation of mastocytosis is ...heterogeneous, ranging from skin-limited disease to more aggressive variants that may be associated with multiorgan dysfunction/failure and shortened survival. In a relatively high proportion of cases, the clonal nature of the disease can be established on the basis of the demonstration of gain-of-function mutations involving the tyrosine kinase (TK) domain of KIT in skin lesions and BM cells and by the factor-independent proliferation and transforming abilities of these mutations. The tyrosine kinase inhibitor Imatinib is a treatment available for mastocytosis patients; however, some KIT mutations, specially KIT D816V, confer resistance to this drug.
To characterize the clinical phenotype and molecular mutations of 2 relatives with diagnosis of systemic mastocytosis (WHO 2008). We also aimed to test the in vitro sensitivity of primary bone marrow (BM) cells from both patients to tyrosine kinase inhibitors.
Four individuals were included in the study; two patients (case 1 mother, and case 2 daughter), and the parents of case 1. DNA samples were obtained from total BM cells, CD3+ BM cells and oral mucosa of patients, and from peripheral blood of all individuals. KIT (exons 1 to 21) was submitted for Sanger sequencing analysis. Primary bone marrow cells (5X104) from the 2 patients were cultured and treated with Imatinib (5uM), Dasatinib (80nM) and PKC 412 (100nM) or with vehicle only (control cells) and submitted for proliferation (MTT) and apoptosis assays (Annexin-V/PI) at days 4, 8 and 12 of culture.
Case 1 was a 33 year-old woman with a chronic history of pruritic skin rash who was referred to our outpatient service for evaluation of massive splenomegaly (25 centimeters in length) and pancytopenia. She had neither comorbidities nor any familial history of hematological malignancies. The patient had no siblings and had only one daughter (case 2). At biopsy, she showed extensive skin and bone marrow infiltration by mast cells. During follow up, the patient presented with spontaneous splenic rupture and had to undergo splenectomy, which led to the resolution of pancytopenia. She was diagnosed with Aggressive Systemic Mastocytosis. Her daughter (case 2), a 17 year-old woman, was also evaluated for an insidious history of diffuse skin rash. Skin and bone marrow biopsies showed massive infiltration by atypical mast cells and a diagnosis of Indolent Systemic Mastocytosis was made. The rare KIT K509I mutation was found in all DNA samples obtained from both patients, but not from the parents of case 1. This suggests that the KIT K509I was a germ line mutation acquired de novo by patient 1 that was subsequently transmitted to her daughter (patient 2). In vitro treatment of primary bone marrow cells harboring the KIT K509I mutation from patients 1 and 2 resulted in variable clinical response rates according to the drug used and the treatment duration. Imatinib treatment resulted in a significant reduction in proliferation (days 4, 8 and 12 of culture) and an increase in apoptosis (days 8 and 12) in cases 1 and 2 (all p≤0.03). Although Dasatinib resulted in decreased proliferation in both patients at day 12 (all p≤0.008), a significantly higher apoptosis ratio was observed only for patient 1 at day 12 of culture (p=0.03). PKC412 had a negative effect over cell growth in patient 1 (days 4 and 8) and in patient 2 (day 4) (all p≤0.03); however, no effect in apoptosis ratio was seen.
We herein provide a report of a KIT K509I mutation in familial mastocytosis. This mutation has been previously described in the literature in one case of familial mastocytosis. Although rare, the screening for KIT K509I mutation should be considered in all cases of familial mastocytosis. Based on in vitro studies, mastocytosis patients harboring the KIT K509I mutation could benefit from treatment with Imatinib, Dasatinib and PKC 412. However, Imatinib may be more effective in inducing neoplastic mast cells apoptosis. Both patients described were started on Imatinib in June 2013.
No relevant conflicts of interest to declare.
PDF
