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Results: 5

1.
Figure 3

Figure 3. Short QT intervals in the Congenic strain compared with S. From: AUGMENTED RIFIFYLIN IS A RISK FACTOR LINKED TO ABERRANT CARDIOMYOCYTE FUNCTION, SHORT QT-INTERVAL AND HYPERTENSION.

Bars ± SEM represent ECG recording once every 5 min continuously for 24 hrs and averaged for 4 hr intervals. (a) Uncorrected short QT interval was observed in the congenic. (b) Representative Electrocardiogram recordings from individual S (n=3) and S.LEW×12×2×3×5 Congenic (n=3) rats. (c) Corrected QT interval (QTcb) ± SEM (by Bazett method). ** p<0.01, *p<0.05

Kathirvel Gopalakrishnan, et al. Hypertension. ;57(4):764-771.
2.
Figure 1

Figure 1. Mapping the blood pressure locus on rat chromosome 10. From: AUGMENTED RIFIFYLIN IS A RISK FACTOR LINKED TO ABERRANT CARDIOMYOCYTE FUNCTION, SHORT QT-INTERVAL AND HYPERTENSION.

(a) The LOD plot for BP using the F2 (S × LEW) population on rat chromosome 10 (RNO10) is followed by the previously published BP QTL containing the four genes shown by the arrows marked as Lig3, LOC688779, Rffl and Rad51l3. The region containing Rffl in red is the current mapped location. The homologous segment on human chromosome 17 (HSA17) is also shown. Numbers alongside RNO10 and HSA17 represent base pair locations.

Kathirvel Gopalakrishnan, et al. Hypertension. ;57(4):764-771.
3.
Figure 4

Figure 4. Expression analysis of Rififylin and Levels of polyubiquitinated proteins. From: AUGMENTED RIFIFYLIN IS A RISK FACTOR LINKED TO ABERRANT CARDIOMYOCYTE FUNCTION, SHORT QT-INTERVAL AND HYPERTENSION.

(a) Expression of Rffl transcript at 53 days of age as detected by RT-PCR. (b) Quantification of Rffl transcripts by Real time PCR (n=6 animals/group). (c) Immunoblot of Rffl in whole-cell lysates from S (n=3) and congenic (n=3) rat hearts at 53 days of age. 36.41kDa, RFFL (NP_0010717368, 2aa–99aa) partial recombinant protein was used as positive control and β-Actin was the loading control. Quantification of Rffl protein± SEM is shown on the right. **p<0.01. (d) Immunoblot for polyubiquitinated proteins in whole-cell lysates from S (n=2) and congenic (n=3) rat hearts at 110 days of age. Control: provided by the Pierce Ubiquitination enrichment kit. Quantification of these proteins ± SEM is shown in the right. **p<0.01.

Kathirvel Gopalakrishnan, et al. Hypertension. ;57(4):764-771.
4.
Figure 5

Figure 5. Cardiac functional analysis of Rififylin. From: AUGMENTED RIFIFYLIN IS A RISK FACTOR LINKED TO ABERRANT CARDIOMYOCYTE FUNCTION, SHORT QT-INTERVAL AND HYPERTENSION.

(a) Differential expression of Rffl gene transcripts from neonatal S.LEW×12×2×3×5 congenic rat hearts (n= 6/group) by Real-time PCR. (b) Defective transferrin recycling in the congenic cardiomyocytes. The disappearance of fluorescently labeled transferrin was plotted using the initial mean intensity of labeled internalized transferrin (± SEM) by the cardiomyocytes taken as 100% in three independent experiments conducted in duplicates. (c) Representative images of data presented in panel (b) from cardiomyocytes recycling transferrin. Green, Alexa488-Transferrin; Blue, DRAQ5. (d) Differential rates of beats/min of neonatal cardiomyocytes. Experiments were conducted twice and beats counted three times for each experiment. *** p<0.001, ** p<0.01

Kathirvel Gopalakrishnan, et al. Hypertension. ;57(4):764-771.
5.
Figure 2

Figure 2. Time course assessment of cardiac function and blood pressure. From: AUGMENTED RIFIFYLIN IS A RISK FACTOR LINKED TO ABERRANT CARDIOMYOCYTE FUNCTION, SHORT QT-INTERVAL AND HYPERTENSION.

(a) Mean systolic BP effect ± SEM by the tail-cuff method of 110 day old rats; S (n=10) and S.LEW×12×2×3×5 congenic (n=10) rats. (b) Echocardiographic measurements were obtained on 110 day old S and congenic rats. Two dimensional images represent the end diastole from S rat and congenic rat. M-mode images from S rat and S.LEW×12×2×3×5 congenic rat. (c) BP data corroborated by radiotelemetry using C40 transmitters. S (n=13) and S.LEW×12×2×3×5 congenic (n=13) rats. Data plotted is the recording obtained once every 5 min continuously for 24hrs and averaged for 4 hr intervals. P values indicated are obtained by two-way ANOVA followed by Tukey’s test (d) Time course assessment of BP by radiotelemetry. The data shown are from 5 rats/group on a low (0.3% NaCl) diet, implanted with the C10 transmitters. Data plotted was obtained by telemetry recording once every 5 min continuously for 24–72 hrs and averaged for 4 hr intervals. Levels of statistical significance by two-way ANOVA followed by Tukey’s test are indicated on the x-axis as follows: ***p<0.001, **p<0.01, *p<0.05. (e) Kaplan-Meier survival curves. Animals (S and S.LEW×12×2×3×5 congenic rats, n=12/group) were fed with 0.3% dietary salt (NaCl) for 116 days and then fed with 2% NaCl until their natural death (p<0.0007).

Kathirvel Gopalakrishnan, et al. Hypertension. ;57(4):764-771.

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