Evaluation of the presence of gingivitis as confounding factor in assessing inflammatory status in serum and saliva of dogs with diabetes mellitus.

The aim of this study was to evaluate the changes in the serum and salivary inflammatory markers induced by Diabetes mellitus (DM) in dogs and to assess the possible confounding effect of gingivitis. A panel of 13 cytokines was measured in the serum and saliva of dogs diagnosed with DM and compared with healthy dogs without gingivitis (control group 1; CG1) and dogs with gingivitis but otherwise healthy (control group 2; CG2). The results of the present study showed statistically significantly higher levels of IL-8, KC-like and MCP1 in the serum of dogs with DM compared to CG1 dogs. In the case of saliva, the DM group presented statistically higher GM-CSF, IL6, IL15, and MCP1 levels compared to CG1, and lower KC-like chemokine compared to CG2. Finally, gingivitis produced changes in saliva, with salivary levels of GM-CSF, IL-6, IL-7, IL-15, IP-10, KC-like, IL-10, IL-18, MCP1, TNFα being statistically significantly higher in the saliva of CG2 dogs compared to CG1. The results of the present study indicate that dogs with DM have altered cytokine levels in serum and saliva compared to healthy dogs. In addition, this study highlights the importance of taking oral health into account when determining cytokines in dogs, as gingivitis can significantly alter their concentrations. .

Corn kernel hardness, drying temperature and amylase supplementation affect live performance and nutrient utilization of broilers.

Drying temperature (DT) of corn can influence its nutritional quality, but whether this is influenced by endosperm hardness is not clear. Two parallel experiments were conducted to investigate the effects of 2 yellow dent corn hybrids with average and hard kernel hardness, dried at 3 temperatures (35, 80, and 120°C), and 2 supplementation levels of an exogenous amylase (0, 133 g/ton of feed) on live performance, starch and protein digestibility, and energy utilization of Ross 708 male broilers. Twelve dietary treatments consisting of a 2 × 3 × 2 factorial arrangement were evaluated using 3-way ANOVA in a randomized complete block design. In Experiment 1, a total of 1,920 male-chicks were randomly allocated to 96 floor pens, whereas 480 day-old chicks were distributed among 96 cages for Experiment 2. At 40 d, interaction effects (P < 0.05) were detected on BWG, FCR, and flock uniformity. Supplementation with exogenous amylase resulted in heavier broilers, better FCR and flock uniformity, only in the diets based on corn dried at 35°C. Additionally, interaction effects were observed on FCR due to kernel hardness and DT (P < 0.01), kernel hardness and amylase supplementation (P < 0.001), and DT and amylase supplementation (P < 0.05). Exogenous amylase addition to the diets based on corn with an average hardness improved FCR up to 2 points (1.49 vs. 1.51 g:g) whereas there was no effect of amylase on FCR of broilers fed diets based on corn with hard endosperm. Total tract retention of starch was increased (P < 0.05) in broilers fed diets based on corn with average kernel hardness compared to hard kernel. Corn dried at 80 and 120°C had up to 1.21% points less starch total tract retention than the one dried at 35°C. Supplementing alpha-amylase resulted in beneficial effects for broiler live performance, energy utilization, and starch total tract digestibility results. Treatment effects on starch characteristics were explored. Corn endosperm hardness, DT and exogenous amylase can influence the live performance of broilers. However, these factors are not independent and so must be manipulated strategically to improve broiler performance.

Effects of amino acid levels during rearing on Cobb 500 slow-feathering broiler breeders: 2. Reproductive performance.

Crude protein and amino acid (AA) content in rearing diets affect body composition and reproductive performance. This study evaluated the effects of 4 dietary AA levels during rearing on BW, egg production and composition, fertility, hatchability, and embryo mortality up to 65 wk of age on Cobb 500 slow-feathering (SF) broiler breeders. The treatments consisted in 80% (low-AA), 90% (moderate-AA), 100% (standard-AA), and 110% (high-AA) of the AA recommendations for Cobb 500 SF pullets from 5 to 24 wk. AA was guided by an ideal protein profile based on digestible Lys. A total of 1,360 pullets and 288 Cobb MV cockerels were randomly placed in 16 pullets and 16 cockerel floor-pens. At 22 wk, 1,040 females and 112 males were transferred into 16-floor pens in a laying house. BW increased linearly (P < 0.01) as AA augmented at 25, 36, and 40 wk. No effects (P > 0.05) at the onset of lay were observed. Moderate-AA and standard-AA resulted in the best hen-housed egg production (HHEP) at 65 wk with 174.3 and 176.5 eggs, respectively. The optimum level of AA for HHEP at 65 wk was estimated (P < 0.001) in 96.7% and 94.7% by the quadratic and broken line models, respectively. Overall, the lightest egg weight (P = 0.022) was obtained with 89%AA during rearing, and the heaviest eggs (P < 0.001) were found at 54 wk. Response surface regression indicated linear effects on albumen and yolk percentages (P < 0.01) increasing and decreasing, respectively, as AA levels augmented; consequently, AA had a negative linear effect on Y:A ratio (P = 0.004) with quadratic effects (P < 0.01) of age (R2 = 0.92). No statistical effect of treatments was observed in fertility (P > 0.05), but AA had a quadratic effect (P = 0.046) on hatchability up to 50 wk of age with 97% as optimum, and decreased linearly (P = 0.004) from 51 to 65 wk. A few effects of treatments (P < 0.05) on embryo mortality were observed. In conclusion, AA levels during rearing affect broiler breeder reproductive performance.

Central and peripheral nervous system involvement by COVID-19: a systematic review of the pathophysiology, clinical manifestations, neuropathology, neuroimaging, electrophysiology, and cerebrospinal fluid findings.

BACKGROUND: SARS-CoV-2 can affect the human brain and other neurological structures. An increasing number of publications report neurological manifestations in patients with COVID-19. However, no studies have comprehensively reviewed the clinical and paraclinical characteristics of the central and peripheral nervous system's involvement in these patients. This study aimed to describe the features of the central and peripheral nervous system involvement by COVID-19 in terms of pathophysiology, clinical manifestations, neuropathology, neuroimaging, electrophysiology, and cerebrospinal fluid findings. METHODS: We conducted a comprehensive systematic review of all the original studies reporting patients with neurological involvement by COVID-19, from December 2019 to June 2020, without language restriction. We excluded studies with animal subjects, studies not related to the nervous system, and opinion articles. Data analysis combined descriptive measures, frequency measures, central tendency measures, and dispersion measures for all studies reporting neurological conditions and abnormal ancillary tests in patients with confirmed COVID-19. RESULTS: A total of 143 observational and descriptive studies reported central and peripheral nervous system involvement by COVID-19 in 10,723 patients. Fifty-one studies described pathophysiologic mechanisms of neurological involvement by COVID-19, 119 focused on clinical manifestations, 4 described neuropathology findings, 62 described neuroimaging findings, 28 electrophysiology findings, and 60 studies reported cerebrospinal fluid results. The reviewed studies reflect a significant prevalence of the nervous system's involvement in patients with COVID-19, ranging from 22.5 to 36.4% among different studies, without mortality rates explicitly associated with neurological involvement by SARS-CoV-2. We thoroughly describe the clinical and paraclinical characteristics of neurological involvement in these patients. CONCLUSIONS: Our evidence synthesis led to a categorical analysis of the central and peripheral neurological involvement by COVID-19 and provided a comprehensive explanation of the reported pathophysiological mechanisms by which SARS-CoV-2 infection may cause neurological impairment. International collaborative efforts and exhaustive neurological registries will enhance the translational knowledge of COVID-19's central and peripheral neurological involvement and generate therapeutic decision-making strategies. REGISTRATION: This review was registered in PROSPERO 2020 CRD42020193140 Available from: https://www.crd.york.ac.uk/prospero/display_record.php?ID=CRD42020193140.

Corn drying temperature, particle size, and amylase supplementation influence growth performance, digestive tract development, and nutrient utilization of broilers.

Broiler live performance may be influenced by postharvest corn drying temperature, and results could depend on particle size after grinding. The supplementation with an exogenous amylase may improve performance parameters, but responses to enzymes are also affected by particle size. Two parallel experiments were conducted to evaluate the effects of hard-kernel corn dried at 2 temperatures (35°C and 120°C), ground at 2 particle sizes (coarse or fine), and 3 supplementation levels (0, 133, and 266 g ton-1) of an exogenous amylase on live performance, gastrointestinal organ development, energy utilization, and nutrient digestibility. Twelve dietary treatments resulting from a 2 × 2 × 3 factorial arrangement of drying temperature, particle size, and amylase supplementation were evaluated in both experiments. A total of 1,920 day-old male chicks were randomly allocated to 96 floor pens, while 480 chicks were distributed among 4 battery brooder units. Ileal and fecal samples were collected to determine energy utilization and nutrient digestibility using titanium dioxide as inert marker. At 42 D, organs were collected, and relative weight or length was determined. Data were analyzed using a three-way ANOVA in a randomized complete block design. Feeding fine corn-based diets showed improvements on live performance for both studies. At 40 D, supplementing 266 g ton-1 of amylase improved feed conversion ratio (P < 0.05) by 1 point compared to chickens that consumed nonsupplemented diets and feed with amylase at 133 g ton-1. Broilers fed coarse corn-based diets had heavier gizzard (P < 0.001) and liver (P < 0.05) than chickens that consumed fine corn-based diets. In addition, starch digestibility was improved by amylase (P < 0.05) at 133 g ton-1 and by feeding coarse corn-based diets (P = 0.06). For chicks raised in cages (16 D), AMEn was increased (P < 0.01) by amylase supplementation regardless of its inclusion level. In conclusion, drying temperature and particle size interactions influenced broiler live performance, gastrointestinal organ development, nutrient digestibility, and energy utilization, and these parameters were improved by supplementing amylase.

A SMAD1/5-YAP signalling module drives radial glia self-amplification and growth of the developing cerebral cortex.

The growth and evolutionary expansion of the cerebral cortex are defined by the spatial-temporal production of neurons, which itself depends on the decision of radial glial cells (RGCs) to self-amplify or to switch to neurogenic divisions. The mechanisms regulating these RGC fate decisions are still incompletely understood. Here, we describe a novel and evolutionarily conserved role of the canonical BMP transcription factors SMAD1/5 in controlling neurogenesis and growth during corticogenesis. Reducing the expression of both SMAD1 and SMAD5 in neural progenitors at early mouse cortical development caused microcephaly and an increased production of early-born cortical neurons at the expense of late-born ones, which correlated with the premature differentiation and depletion of the pool of cortical progenitors. Gain- and loss-of-function experiments performed during early cortical neurogenesis in the chick revealed that SMAD1/5 activity supports self-amplifying RGC divisions and restrains the neurogenic ones. Furthermore, we demonstrate that SMAD1/5 stimulate RGC self-amplification through the positive post-transcriptional regulation of the Hippo signalling effector YAP. We anticipate this SMAD1/5-YAP signalling module to be fundamental in controlling growth and evolution of the amniote cerebral cortex.

E proteins sharpen neurogenesis by modulating proneural bHLH transcription factors' activity in an E-box-dependent manner.

Class II HLH proteins heterodimerize with class I HLH/E proteins to regulate transcription. Here, we show that E proteins sharpen neurogenesis by adjusting the neurogenic strength of the distinct proneural proteins. We find that inhibiting BMP signaling or its target ID2 in the chick embryo spinal cord, impairs the neuronal production from progenitors expressing ATOH1/ASCL1, but less severely that from progenitors expressing NEUROG1/2/PTF1a. We show this context-dependent response to result from the differential modulation of proneural proteins' activity by E proteins. E proteins synergize with proneural proteins when acting on CAGSTG motifs, thereby facilitating the activity of ASCL1/ATOH1 which preferentially bind to such motifs. Conversely, E proteins restrict the neurogenic strength of NEUROG1/2 by directly inhibiting their preferential binding to CADATG motifs. Since we find this mechanism to be conserved in corticogenesis, we propose this differential co-operation of E proteins with proneural proteins as a novel though general feature of their mechanism of action.

Measurement of salivary adiponectin concentrations in dogs.

BACKGROUND: Measurement of salivary adiponectin could improve understanding of this adipokine's physiology, and its role in various clinical conditions. OBJECTIVES: The purpose of the study was to evaluate the utility of a human adiponectin ELISA kit for measurement of salivary adiponectin in dogs, to compare serum and salivary adiponectin concentrations in a healthy dog population, and to evaluate possible effects of tooth-cleaning on serum and salivary adiponectin concentrations in dogs. METHODS: For analytical validation, precision, accuracy, and lower limit of quantification of the assay were determined with saliva samples. In addition, adiponectin concentrations were quantified in serum and saliva samples from 24 healthy dogs, and from 7 dogs with mild gingivitis before and after a tooth-cleaning procedure. RESULTS: The validation assays for salivary adiponectin had all coefficients of variation <15%, and recovery ranged from 85% to 120%. In the linearity test, interference was observed when measuring adiponectin in saliva, but this was solved by diluting samples 1:4. In healthy dogs, salivary and serum adiponectin concentrations were positively correlated (r = .650; P = .009). After the tooth-cleaning procedure, salivary adiponectin concentration increased on day 0 (P = .004), but by day 14, concentrations were less than prior to the procedure (P = .041). CONCLUSIONS: The human adiponectin ELISA kit can be used for precise and accurate salivary adiponectin measurement in dogs. Salivary adiponectin increased 24 hours after tooth-cleaning, possibly due to acute inflammation or adiponectin leakage from the blood after gingival trauma.

Canine demodicosis: the relationship between response to treatment of generalised disease and markers for inflammation and oxidative status.

BACKGROUND: Determination of acute phase proteins and markers of oxidative status may be of value for evaluating the severity of disease and the response to treatment. In canine demodicosis, there is no information available regarding the use of such analytes to discriminate between localized and generalized demodicosis or to monitor the response to treatment. HYPOTHESIS/OBJECTIVES: The aim was to measure analytes related to inflammation and oxidative stress in dogs with localized or generalized demodicosis. In cases of generalized demodicosis, the intention was to study these analytes before and after a period of treatment. ANIMALS: Serum was obtained from three groups: Group 1, healthy dogs; Group 2, dogs with localized demodicosis; and Group 3, dogs with generalized demodicosis. METHODS: Animals from Groups 1 and 2 were sampled at the point of diagnosis. Dogs in Group 3 were treated with oral ivermectin 1% at 0.6 mg/kg once daily, and samples were collected at the point of diagnosis and after 30 days of treatment. C-Reactive protein, haptoglobin, albumin, butyrylcholinesterase, paraoxonase-1 and total antioxidant capacity were measured. RESULTS: Dogs with generalized demodicosis had significantly higher concentrations of C-reactive protein and haptoglobin and lower butyrylcholinesterase activity than dogs in Groups 1 and 2. Dogs in Group 3 also had lower paraoxonase-1 than those in Group 2, The analytes tended to normalize during treatment. CONCLUSIONS AND CLINICAL IMPORTANCE: There was an evident acute phase response and changes in selected oxidative state analytes in generalized demodicosis that do not occur in the localized form. These changes could be used for monitoring the response to treatment.

Urinary C reactive protein levels in dogs with leishmaniasis at different stages of renal damage.

The objectives of the study were to validate a time-resolved immunofluorometric assay for C reactive protein (CRP) quantification in urine of dogs and to investigate the influence that the presence of proteinuria and azotemia could have on serum and urinary CRP (uCRP) values in dogs with leishmaniasis. Samples obtained from dogs naturally infected with Leishmania infantum were classified into four groups on the basis of the results of urinary protein/creatinine ratio and serum creatinine (sCr). In addition, 7 dogs were monitored at initial diagnosis and after a follow up visit. The assay showed good analytical performance based on precision, accuracy and limit of detection results. Results of the study suggested that CRP is present in urine of dogs with leishmaniasis and renal damage since uCRP/creatinine ratio was significantly increased in dogs with proteinuria, being the highest values observed in dogs with proteinuria and elevated sCr, and that the measurement of uCRP could be a tool to detect and evaluate the possible kidney damage associated with this disease.

Appropriate timing of nitroglycerin prior to intravascular ultrasound.

OBJECTIVES: To determine the time to maximal coronary dilation following intracoronary (IC) nitroglycerin (NTG) and whether the decrease in aortic pressure (AoP) is a surrogate marker for coronary vasodilatation. BACKGROUND: Intravascular ultrasound (IVUS) facilitates assessment of coronary plaque severity and morphology and aids in stent sizing. NTG is often administered prior to IVUS to prevent catheter-induced spasm and to facilitate standardized and accurate vessel size measurements. The impact of dose, timing, and route of delivering NTG on vessel size remains undefined. METHODS: Twelve patients undergoing IVUS-guided stent placement were studied. An IVUS catheter was positioned proximal to the target lesion and the following measurements made at baseline and 30 second (sec) intervals for 180 sec following 200 mcg IC NTG: AoP, IVUS-derived lumen diameter (Ld), lumen cross-sectional area (La), external elastic membrane diameter (EEMd) and EEM area (EEMa). Lumen and EEM measurements were compared at different time intervals and the relationship between time to max Ld and nadir AoP was analyzed. RESULTS: All patients had a vasodilatory response to IC NTG. Increase from baseline to max Ld following IC NTG was statistically significant (mean change 0.31 ± 0.18 mm, P=.0001). Mean time to max Ld following IC NTG was 117 sec (range, 60-180 sec). No correlation between time to max Ld and AoP nadir was observed (r = 0.19). CONCLUSIONS: Our study suggests that administration of 200 mcg IC NTG results in a significant change in lumen diameter and area with maximal vasodilation occurring on average approximately 2 minutes following IC NTG administration. There was no significant correlation between AoP change and maximal NTG-induced coronary vasodilation.

Urinary clusterin as a renal marker in dogs.

A validation of a species-specific enzyme immunoassay for urinary clusterin measurement in dogs was performed, and the use of urinary clusterin as a marker of renal damage was evaluated in a population of dogs with leishmaniasis. Urine was obtained from 75 dogs; 64 dogs had leishmaniasis and 11 were healthy. The dogs with leishmanias were divided into 5 groups: I (n = 9; serum creatinine [SCr] < 1.4 mg/dl, urinary protein-to-creatinine [UPC] ratio ≤ 0.5); II (n = 29; SCr < 1.4 mg/dl, UPC > 0.5); III (n = 6; SCr ≥ 1.4 mg/dl to <2 mg/dl, UPC > 0.5); IV (n = 13; SCr ≥ 2 mg/dl to <5 mg/dl, UPC > 0.5); and V (n = 7; SCr ≥ 5 mg/dl, UPC > 0.5). The urinary clusterin concentration was measured, and the urinary clusterin-to-creatinine ratio was calculated. Canine urinary clusterin assay showed good analytical performance based on precision accuracy and limit-of-detection results. There was a statistically significant increase in urinary clusterin and clusterin-to-creatinine ratio in groups II-V compared with group I and healthy group. The results of the current study showed that urinary clusterin concentration and urinary clusterin-to-creatinine ratios are increased in dogs with analytical evidences of renal damage and that the urinary clusterin-to-creatinine ratio might be used as a potential early biomarker of chronic kidney disease.

Acute phase protein response in the capybara (Hydrochoerus hydrochaeris).

We evaluated the acute phase protein response in capybaras (Hydrochoerus hydrochaeris). Three animal groups were used: 1) healthy animals (n=30), 2) a group in which experimental inflammation with turpentine was induced (n=6), and 3) a group affected with sarcoptic scabies (n=14) in which 10 animals were treated with ivermectin. Haptoglobin (Hp), acid-soluble glycoprotein (ASG) and albumin were analyzed in all animals. In those treated with turpentine, Hp reached its maximum value at 2 wk with a 2.7-fold increase, whereas ASG increased 1.75-fold and albumin decreased 0.87-fold 1 wk after the induction of inflammation. Capybaras affected with sarcoptic scabies presented increases in Hp and ASG of 4.98- and 3.18-fold, respectively, and a 0.87-fold decrease in albumin, compared with healthy animals. Haptoglobin and ASG can be considered as moderate, positive acute phase proteins in capybaras because they showed less than 10-fold increases after an inflammatory process and reached their peak concentrations 1 wk after the induction of inflammation. Conversely, albumin can be considered a negative acute phase protein in capybaras because it showed a reduction in concentration after inflammatory stimulus.

Canine C-reactive protein measurements in cerebrospinal fluid by a time-resolved immunofluorimetric assay.

In the current study, the quantification of C-reactive protein (CRP) in cerebrospinal fluid (CSF) of dogs using an adapted time-resolved immunofluorimetric assay (TR-IFMA) was investigated, as well as whether the assay could be used to detect the range of CRP concentrations found in different clinical situations. Intra- and interassay coefficients of variation were below 15% in all cases. The TR-IFMA measured the CRP values in a proportional and linear manner (r  =  0.99); also CRP concentrations measured in CSF and in serum were significantly correlated (r  =  0.80, P  =  0.003). The limit of detection of the method was 7.1 × 10(-6) mg/l. The assay was able to detect differences in CRP concentrations in CSF of dogs with inflammatory disorders compared with dogs with spinal cord compression or idiopathic epilepsy. In conclusion, TR-IFMA constitutes a very sensitive, precise, and accurate method for the measurement of CRP concentrations in CSF.

Regulation of aurora B kinase by the lipid raft protein flotillin-1.

The lipid raft protein Flotillin-1 was previously shown to be required for cell proliferation. Here we show that it is critical for the maintenance of the levels of the mitotic regulator Aurora B. Knockdown of Flotillin-1 induced aberrant mitotic events similar to those produced by Aurora B depletion and led to a marked decline in Aurora B levels and activity. Transfection of wild-type full-length Flotillin-1 or forms directed to the nucleus increased Aurora B levels and activity. Flotillin-1 interacted with Aurora B directly through its SPFH domain in a complex distinct from the chromosomal passenger protein complex, and the two proteins co-purified in nuclear, non-raft fractions. These observations are the first evidence for a function of Flotillin-1 outside of lipid rafts and suggest its critical role in the maintenance of a pool of active Aurora B.

Hypothyroidism and renal function in patients with systolic heart failure.

The extent to which hypothyroidism affects renal function in patients with heart failure remains incompletely explored, despite the known adverse prognostic implications of renal dysfunction in these patients.In a pilot retrospective study, we evaluated 75 patients (age, >or=18 yr) with left ventricular ejection fractions <0.40. Forty-five patients had normal thyroid function (thyroid-stimulating hormone [TSH], 0.35-5.5 micro IU/mL) and 30 had hypothyroidism. The group with hypothyroidism was subdivided into 17 patients who had controlled hypothyroidism (TSH, 0.35-5.5 micro IU/mL) and 13 who had uncontrolled hypothyroidism (TSH, >5.5 micro IU/mL). Renal function, measured in terms of glomerular filtration rate, was analyzed once in each patient, and the populations were statistically compared, with P <0.05 conferring statistical significance.Baseline characteristics in all groups were similar. Mean glomerular filtration rate was better in patients with normal thyroid function than those with hypothyroidism (75.45 +/- 31.48 vs 63.95 +/- 21.43 mL/min/1.73 m2; P=0.032). There was no significant difference between patients with controlled hypothyroidism (66.89 +/- 24.18 mL/min/1.73 m2) and those with normal thyroid function (P=0.131). In patients with uncontrolled hypothyroidism, mean glomerular filtration rate (60.2 +/- 17.4 mL/min/1.73 m2) was significantly worse than in patients with normal thyroid function (P=0.015).We found that heart-failure patients with insufficiently treated hypothyroidism have worse renal function than do patients whose thyroid function is normal or whose hypothyroidism is effectively treated. Larger studies will be needed in order to evaluate this conclusion further. We recommend that hypothyroidism in heart-failure patients be strictly controlled, lest it affect prognosis adversely.

Escherichia coli septic shock masquerading as ST-segment elevation myocardial infarction.

Elevated cardiac biomarkers in conjunction with electrocardiographic (ECG) changes are valuable in diagnosing acute coronary syndrome (ACS). Elevated troponin I (TnI), while commonly seen in ACS, can also occur in entities such as sepsis and pulmonary thromboembolic disease. Raised TnI levels in patients with sepsis result from various mechanisms, including hypoperfusion or direct extension of infection to cardiac tissue, and can also serve as an important prognostic indicator. Electrocardiographic changes in sepsis are not as well described. Some of the ECG findings associated with septic shock include loss of QRS amplitude, increase in QTc interval, bundle branch blocks, and development of narrowed QRS intervals with deformed, positively deflected J waves (commonly known as Osborn waves). ST-segment elevations in sepsis are rare and have only previously been noted in a handful of case reports involving patients with septic shock. We present a case of a 59-year-old woman with ST-segment elevations and increased levels of cardiac troponin from Escherichia coli septic shock in the setting of normal coronary angiography.

The role of eNOS, iNOS, and NF-kappaB in upregulation and activation of cyclooxygenase-2 and infarct size reduction by atorvastatin.

Pretreatment with atorvastatin (ATV) reduces infarct size (IS) and increases myocardial expression of phosphorylated endothelial nitric oxide synthase (p-eNOS), inducible NOS (iNOS), and cyclooxygenase-2 (COX2) in the rat. Inhibiting COX2 abolished the ATV-induced IS limitation without affecting p-eNOS and iNOS expression. We investigated 1) whether 3-day ATV pretreatment limits IS in eNOS(-/-) and iNOS(-/-) mice and 2) whether COX2 expression and/or activation by ATV is eNOS, iNOS, and/or NF-kappaB dependent. Male C57BL/6 wild-type (WT), University of North Carolina eNOS(-/-) and iNOS(-/-) mice received ATV (10 mg.kg(-1).day(-1); ATV(+)) or water alone (ATV(-)) for 3 days. Mice underwent 30 min of coronary artery occlusion and 4 h of reperfusion, or hearts were harvested and subjected to ELISA, immunoblotting, biotin switch, and electrophoretic mobility shift assay. As a result, ATV reduced IS only in the WT mice. ATV increased eNOS, p-eNOS, iNOS, and COX2 levels and activated NF-kappaB in WT mice. It also increased myocardial COX2 activity. In eNOS(-/-) mice, ATV increased COX2 expression but not COX2 activity or iNOS expression. NF-kappaB was not activated by ATV in the eNOS(-/-) mice. In the iNOS(-/-) mice, eNOS and p-eNOS levels were increased but not iNOS and COX2 levels; however, NF-kappaB was activated. In conclusion, both eNOS and iNOS are essential for the IS-limiting effect of ATV. The expression of COX2 by ATV is iNOS, but not eNOS or NF-kappaB, dependent. Activation of COX2 is dependent on iNOS.

Aspirin before reperfusion blunts the infarct size limiting effect of atorvastatin.

We assessed whether aspirin (acetylsalicylic acid, ASA), administered before reperfusion, abrogates the infarct size (IS)-limiting effect of atorvastatin (ATV). Statins reduce IS. This dose-dependent effect is mediated by upregulation of cycloxygenase-2 (COX2) and PGI(2) production. Administration of selective COX2-inhibitors either with ATV for 3 days or immediately before coronary occlusion blocks the IS-limiting effect of ATV. Sprague-Dawley rats received 3-day ATV (10 mg x kg(-1) x day(-1)) or water alone. Rats underwent 30 min coronary artery occlusion and 4 h reperfusion (IS protocol, n=8 in each group), or rats underwent 30 min coronary artery occlusion and 10 min reperfusion (enzyme expression and activity protocol, n=4 in each group). Immediately before reperfusion rats received intravenous ASA (5, 10, or 20 mg/kg) or saline. Area-at-risk (AR) was assessed by blue dye and IS by triphenyltetrazolium chloride. ATV reduced IS (10.1 +/- 1.4% of the AR) compared with controls (31.0 +/- 2.2%). Intravenous ASA alone did not affect IS (29.0 +/- 2.6%); however, ASA dose dependently (5, 10, and 20 mg/kg) attenuated the protective effect of ATV on IS (15.8 +/- 0.9%, 22.0 +/- 1.6%, and 23.7 +/- 3.8%, respectively). ASA dose dependently blocked the upregulation of COX2 by ATV. COX2 activity was as follows: control, 8.93 +/- 0.90 pg/mg; ATV, 75.85 +/- 1.08 pg/mg; ATV + ASA5, 34.39 +/- 1.48 pg/mg; ATV + ASA10, 19.87 +/- 1.10 pg/mg; and ATV + ASA20, 9.36 +/- 0.94 pg/mg. ASA, administered before reperfusion in doses comparable to those used in the clinical setting, abrogates the IS-limiting effect of ATV in a model with mechanical occlusion of the coronary artery. This potential adverse interaction should be further investigated in the clinical setting of acute coronary syndromes.