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1 ml; CTL, n = 7; XNT, n = 7; STZ, n = 7; and STZ + XNT, n = 8), in accordance with previous studies (Franchini & Krieger, 1993). The changes in MAP and HR (��MAP, in millimetres of mercury, and ��HR, in beats per minute) were analysed [http://en.wikipedia.org/wiki/Alkaline_phosphatase (-)-p-Bromotetramisole Oxalate] at the peak of the responses. The autonomic function was evaluated using sequential and alternated pharmacological blockage of muscarinic and ��-adrenergic receptors with methyl-atropine (3 mg kg-1, 0.2 ml per rat) and atenolol (8 mg kg-1, 0.2 ml per rat), respectively, with an interval of 15 min between the injections. The intrinsic heart rate (IHR) was evaluated after 15 min of the double blockage with methyl-atropine and atenolol, and it was used for calculation of the parasympathetic (IHR minus minimal HR; CTL, n = 8; XNT, n = 7; STZ, n = 6; and STZ + XNT, n = 7) and sympathetic tones (maximal HR minus IHR; (CTL, n = 8; XNT, n = 7; STZ, n = 6; and STZ + XNT, n = 7; Negrao et al. 1992). At the end of the invasive cardiovascular analysis, [http://www.selleckchem.com/products/ipi-145-ink1197.html INK1197 nmr] the rats (CTL, n = 8; XNT, n = 8; STZ, n = 7; and STZ + XNT, n = 6) were heparinized (400 i.u., i.p.) and decapitated. The thorax was opened, and the heart was carefully dissected, removed and placed in a cold Krebs�CRinger solution (mm: 118.4 NaCl, 4.7 KCl, 1.2 KH2PO4, 1.2 MgSO4.7H2O, 2.5 CaCl2.2H2O, 11.7 glucose and 26.5 NaHCO3) to preserve the heart before the perfusion. As described previously (Goes et al. 1993), the hearts were perfused through the aortic stump with Krebs�CRinger solution at 37 �� 1��C in a Langendorff system with constant pressure (65 mmHg) and oxygenation (5% CO2 and 95% O2). A force transducer (TSD 104 A; Biopac Systems Inc.) was attached through [http://www.selleckchem.com/products/JNJ-26481585.html JNJ-26481585 cost] a heart clip to the apex of the ventricles to record the contractile force using a data-acquisition system (Acqknowledge; Biopac Systems Inc.). A diastolic tension of 1.0 �� 0.2 g was applied to the hearts. Coronary flow was measured by collecting the perfusate over a period of 1 min at regular intervals. Heart rate and ��dT/dt (time derivative of tension) were calculated from the contractile tension recordings. After 30 min of stabilization, the functional parameters were recorded for an additional period of 30 min. Data are expressed as means �� SEM. Statistical analysis was performed using Student's unpaired t test (glycaemia) or one-way ANOVA followed by the Newman�CKeuls post hoc test. A value of P