ABUNDANCE OF YELLOW ALKALOID ‘BERBERINE’ IN THE MEDICINAL PLANT OF TALI KUNING (Tinospora dissitiflora Diels.) COLLECTED FROM MANOKWARI – WEST PAPUA

This research was conducting to quantify the berberine concentration. Rapid method using 1H-NMR was used to determine berberine quantitative and qualitatively from the crude extract. Tali kuning was collected from Manokwari Papua Barat and powdered with hammer mill. Sonication was employed to extract using methanol at room temperature. 500 µg authentic berberine chloride were dissolved in 5.4 mL methanol-d4 (containing 84.4 µg anthracene). 1H-NMR spectra were recorded in methanol-d4 (99,9%) using JEOL JNM-ECX 500. Each sample was scanned for 100 using the following parameter 0.18 Hz/point, spectral width 14400 Hz, pulse with 4.0 US, relaxation delay 2 sc. Peak areas were used for qualitative analysis and integration of each peach were employed for quantitative analysis. The results demonstrated that 1H-NMR signal pattern of H-13 and H-8 recorded from Tali Kuning, and Amur corktree were well recorded, and in accordance to the berberine chloride standard. Using peak integration of H-13 and H-8, the berberine quantity in Tali kuning is 18.06 mg/g of dried powder, and 22.78 mg//gr for Amur corktree. Berberine percentage based on the weight of oven-dried-extracts was 8.34% (MC 7.54%) and 12.04% (11,54%) for Amur corktree and Tali kuning.

Heart failure-associated alterations in troponin I phosphorylation impair ventricular relaxation-afterload and force-frequency responses and systolic function

Recent studies have found that selective stimulation of troponin (Tn)I protein kinase A (PKA) phosphorylation enhances heart rate-dependent inotropy and blunts relaxation delay coupled to increased afterload. However, in failing hearts, TnI phosphorylation by PKA declines while protein kinase C (PKC) activity is enhanced, potentially augmenting TnI PKC phosphorylation. Accordingly, we hypothesized that these site-specific changes deleteriously affect both rate-responsive cardiac function and afterload dependence of relaxation, both prominent phenotypic features of the failing heart. A transgenic (TG) mouse model was generated in which PKA-TnI sites were mutated to mimic partial dephosphorylation (Ser22 to Ala; Ser23 to Asp) and dominant PKC sites were mutated to mimic constitutive phosphorylation (Ser42 and Ser44 to Asp). The two highest-expressing lines were further characterized. TG mice had reduced fractional shortening of 34.7 ± 1.4% vs. 41.3 ± 2.0% ( P = 0.018) and slight chamber dilation on echocardiography. In vivo cardiac pressure-volume studies revealed near doubling of isovolumic relaxation prolongation with increasing afterload in TG animals ( P < 0.001), and this remained elevated despite isoproterenol infusion (PKA stimulation). Increasing heart rate from 400 to 700 beats/min elevated contractility 13% in TG hearts, nearly half the response observed in nontransgenic animals ( P = 0.005). This blunted frequency response was normalized by isoproterenol infusion. Abnormal TnI phosphorylation observed in cardiac failure may explain exacerbated relaxation delay in response to increased afterload and contribute to blunted chronotropic reserve.