The aim of the present study was to determine the prevalence and risk factors concerning Dirofilaria immitis infection in dogs from Figueira da Foz, located in the central region of Portugal. In the period between November 2009 and January 2011, 304 blood samples were obtained from dogs over 1 year of age, with no previous history of heartworm prevention or diagnosis. Every blood sample was analyzed using varied laboratory techniques (direct microscopic evaluation of a fresh blood sample, the modified Knott technique, and the ELISA antigen detection test – IDEXX Snapp®). In the samples in which microfilaremia was detected, a histochemical technique using acid phosphatase staining was applied to identify the species of microfilariae. A total prevalence of 27.3% (83 out of 304) was found. We also found that 73.5% of all positive cases (61 out of 83) were microfilaremic, and 26.5% were occult infections (22 out of 83). By means of a histochemical technique Dirofilaria immitis was identified in 96.7% of microfilaremic samples. A multivariate model allowed us to identify the following risk factors for the presence of heartworm disease: age between 4 and 9 years, dogs living in a rural environment, large breed dogs, and living outdoors. This study shows for the first time the high prevalence of heartworm disease in a central area of Portugal and emphasizes the importance of systematic screening for this disease, as well as the need to prevent it in dogs in this area.
The contribution of radiotherapy, per se, to late cardiotoxicity remains controversial. To clarify its impact on the development of early cardiac dysfunction, we developed an experimental model in which the hearts of rats were exposed, in a fractionated plan, to clinically relevant doses of ionizing radiation for oncological patients that undergo thoracic radiotherapy. Rat hearts were exposed to daily doses of 0.04, 0.3, and 1.2 Gy for 23 days, achieving cumulative doses of 0.92, 6.9, and 27.6 Gy, respectively. We demonstrate that myocardial deformation, assessed by global longitudinal strain, was impaired (a relative percentage reduction of >15% from baseline) in a dose-dependent manner at 18 months. Moreover, by scanning electron microscopy, the microvascular density in the cardiac apex was significantly decreased exclusively at 27.6 Gy dosage. Before GLS impairment detection, several tools (qRT-PCR, mass spectrometry, and western blot) were used to assess molecular changes in the cardiac tissue. The number/expression of several genes, proteins, and KEGG pathways, related to inflammation, fibrosis, and cardiac muscle contraction, were differently expressed in the cardiac tissue according to the cumulative dose. Subclinical cardiac dysfunction occurs in a dose-dependent manner as detected by molecular changes in cardiac tissue, a predictor of the severity of global longitudinal strain impairment. Moreover, there was no dose threshold below which no myocardial deformation impairment was detected. Our findings i) contribute to developing new markers and exploring non-invasive magnetic resonance imaging to assess cardiac tissue changes as an early predictor of cardiac dysfunction; ii) should raise red flags, since there is no dose threshold below which no myocardial deformation impairment was detected and should be considered in radiation-based imaging and -guided therapeutic cardiac procedures; and iii) highlights the need for personalized clinical approaches.
The 13C-isotopomer enrichment of hepatic cytosolic acetyl-CoA of overnight-fed mice whose drinking water was supplemented with [U-13C]fructose, and [1-13C]glucose and p-amino benzoic acid (PABA) was quantified by 13C NMR analysis of urinary N-acetyl-PABA. Four mice were given normal chow plus drinking water supplemented with 5% [1-13C]glucose, 2.5% [U-13C]fructose, and 2.5% fructose (Solution 1) overnight. Four were given chow and water containing 17.5% [1-13C]glucose, 8.75% [U-13C]fructose and 8.75% fructose (Solution 2). PABA (0.25%) was present in both studies. Urinary N-acetyl-PABA was analyzed by 13C NMR. In addition to [2-13C]- and [1,2-13C]acetyl isotopomers from catabolism of [U-13C]fructose and [1-13C]glucose to acetyl-CoA, [1-13C]acetyl was also found indicating pyruvate recycling activity. This precluded precise estimates of [1-13C]glucose contribution to acetyl-CoA while that of [U-13C]fructose was unaffected. The fructose contribution to acetyl-CoA from Solutions 1 and 2 was 4.0 ± 0.4% and 10.6 ± 0.6%, respectively, indicating that it contributed to a minor fraction of lipogenic acetyl-CoA under these conditions.
13 C]acetate during a separate visit in a subset of ND (n ϭ 7) subjects. Ratio of 13 C3/ 13 C4 obtained following either tracer was Ͻ1.0 at baseline and during clamp, indicating that TPI exchange was essentially complete and did not contribute to asymmetric glucose enrichment. Uncorrected and corrected rates of gluconeogenesis were no different (P ϭ not significant) in T2DM vs. ND both at baseline and during clamp. TA correction resulted in equivalent estimates of corrected gluconeogenesis in T2DM and ND that were ϳ25-35% lower than uncorrected gluconeogenesis both at baseline and during the clamp. The asymmetric enrichment of glucose from 13 C-gluconeogenic tracers is attributable to TA exchange and can be utilized to correct for TA exchange. In conclusion, TA exchange does not differ between T2DM and ND under fasting or hyperglycemic clamp conditions, and the 2 H2O method continues to provide an accurate estimation of gluconeogenesis. gluconeogenesis; deuterated water; transaldolase; triose phosphate isomerase AFTER AN OVERNIGHT FAST, plasma glucose is not symmetrically labeled from gluconeogenic carbon tracers such as [U-13 C] glycerol (21) and [3-14 C]lactate (25). Exchange of fructose 6-phosphate and triose phosphate mediated by transaldolase has been implicated by us (3, 7, 9) and others (12) in overestimation of rates of gluconeogenesis, utilizing the deuterated water method. In this method, glucose derived from glycogenolysis (GGL) via glucose 6-phosphate (G6P) source can become labeled in carbons 4, 5, and 6 from either a carbonlabeled gluconeogenic precursor or deuterated water independent of gluconeogenesis. This results in an overestimation of the gluconeogenic fraction and a corresponding underestimation of the glycogenolytic contribution to endogenous glucose production. This exchange is particularly important in the study of type 2 diabetes mellitus (T2DM) since it mimics the characteristic shift toward increased hepatic gluconeogenic activity during the progression of this disease. With a few exceptions (14), stable isotope tracer studies indicate an increased fractional gluconeogenesis in overnight-fasted T2DM subjects compared with healthy nondiabetic (ND) controls (6,10,15,16,28). Transaldolase (TA) exchange activity has been implicated in both ND and T2DM subjects from the depletion of position 5 relative to position 3 label following metabolism of [3, H 2 ]galactose to glucose (7). However, it is not known whether transaldolase exchange activity is different between ND and T2DM under conditions of fasting and hyperglycemia (frequently seen with meals). This study was undertaken to compare TA exchange reaction in ND and T2DM subjects using [1-13 C]acetate infusion under conditions of fasting and during a hyperglycemic moderate-dose insulin clamp to determine whether or not TA exchange activity explains differences in glucose enrichment from gluconeogenic substrates between ND and T2DM subjects.We have demonstrated that TA activity occurs in healthy humans based on a higher enrichment of gluco...
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