Brza i osjetljiva HPLC metoda za određivanje doksorubicina u krvi psa Artifakt srebrovog nitrata

A rapid and sensitive high performance liquid chromatographic (HPLC) assay utilizing fluorimetric detection (excitation at 480 nm, emission at 560 nm) for the determination of doxorubicin in dog blood was developed and validated. Treatment of blood samples containing doxorubicin with AgNO3 (as protein precipitant) resulted in appearance of an additional peak in the chromatogram of doxorubicin at 11.5 min along with the parent peak (tR = 5.5 min). The latter peak was not found when treated with other protein precipitants such as trichloroacetic acid and methanol. Construction of calibration curve based on area of both peaks alone did not result in linearity of the curve. However, summation of areas of both peaks resulted in a curve with good linearity and correlation coefficient (R2 = 0.9985). Appearance of second peak may be due to the interaction of doxorubicin with cellular components of blood in the presence of AgNO3 leading to the formation of complex with reduced polarity. Analysis of the quality control samples showed good accuracy (96.7-100.42) and precision (RSD = 2,6-5,7%). The proposed method could be advantageous in estimation of doxorubicin incorporated into targeted delivery systems that concentrate in blood cells and quantify the absolute blood concentration of doxorubicin.Razvijena je i validirana brza i osjetljiva metoda visokotlačne tekućinske kromatografije (HPLC) s fluorimetrijskom detekcijom za određivanje doksorubicina u krvi psa. Nakon obrade uzoraka krvi koji sadrže doksorubicin s AgNO3 (taložno sredstvo za proteine), uz osnovni signal (tR = 5,5min) pojavljuje se dodatni signal u kromatogramu doksorubicina na 11,5 min. Nikakav dodatni signal se ne pojavljuje ako se za taloženje proteina upotrijebe trikloroctena kiselina ili metanol. Međutim, u hemoliziranim uzorcima plazme kojima je dodan doksorubicin nakon obrade sa srebrovim nitratom pojavljuje se drugi signal. Kalibracijska krivulja s površinom ispod signala za samo prvi pik (tR = 5,5min) nije linearna. Međutim, suma površina ispod prvog (tR = 5.5min) i drugog signala (tR = 11.5min) daje linearnu zavisnost s korelacijskim koeficijentom R2 = 0,9985. Pojava drugog signala mogla bi biti posljedica interakcije doksorubicina sa staničnim komponentama u krvi u prisutnosti AgNO3 te nastajanja kompleksa smanjene polarnosti. Površina ispod signala na kromatogramu određena na temelju fluorescencijske detekcije na 480 nm i 560 nm (valne duljine pobuđivanja, odnosno emisije) upotrebljena je za kvantitativnu analizu doksorubicina. Određivanje doksorubicina u krvi psa je jednostavna, precizna i točna. U svim eksperimentima, relativna standardna devijacija (RSD) najčešće je bila < 10%, a izmjerena i teorijska koncentracija razlikovala se za < 10%. Doksorubicin inkubiran s uzorcima krvi psa bio je stabilan najmanje 3 dana ako je čuvan na 5 °C, odnosno najmanje 8 h ako je čuvan na sobnoj temperaturi (28 °C). Predložena HPLC metoda mogla bi imati prednosti u određivanju doksorubicina u sustavima za isporuku lijekova ciljano u krvne stanice i kvantifikaciji ukupne koncentracije doksorubicina u krvi

Brza i osjetljiva HPLC metoda za određivanje doksorubicina u krvi psa Artifakt srebrovog nitrata

A rapid and sensitive high performance liquid chromatographic (HPLC) assay utilizing fluorimetric detection (excitation at 480 nm, emission at 560 nm) for the determination of doxorubicin in dog blood was developed and validated. Treatment of blood samples containing doxorubicin with AgNO3 (as protein precipitant) resulted in appearance of an additional peak in the chromatogram of doxorubicin at 11.5 min along with the parent peak (tR = 5.5 min). The latter peak was not found when treated with other protein precipitants such as trichloroacetic acid and methanol. Construction of calibration curve based on area of both peaks alone did not result in linearity of the curve. However, summation of areas of both peaks resulted in a curve with good linearity and correlation coefficient (R2 = 0.9985). Appearance of second peak may be due to the interaction of doxorubicin with cellular components of blood in the presence of AgNO3 leading to the formation of complex with reduced polarity. Analysis of the quality control samples showed good accuracy (96.7-100.42) and precision (RSD = 2,6-5,7%). The proposed method could be advantageous in estimation of doxorubicin incorporated into targeted delivery systems that concentrate in blood cells and quantify the absolute blood concentration of doxorubicin.Razvijena je i validirana brza i osjetljiva metoda visokotlačne tekućinske kromatografije (HPLC) s fluorimetrijskom detekcijom za određivanje doksorubicina u krvi psa. Nakon obrade uzoraka krvi koji sadrže doksorubicin s AgNO3 (taložno sredstvo za proteine), uz osnovni signal (tR = 5,5min) pojavljuje se dodatni signal u kromatogramu doksorubicina na 11,5 min. Nikakav dodatni signal se ne pojavljuje ako se za taloženje proteina upotrijebe trikloroctena kiselina ili metanol. Međutim, u hemoliziranim uzorcima plazme kojima je dodan doksorubicin nakon obrade sa srebrovim nitratom pojavljuje se drugi signal. Kalibracijska krivulja s površinom ispod signala za samo prvi pik (tR = 5,5min) nije linearna. Međutim, suma površina ispod prvog (tR = 5.5min) i drugog signala (tR = 11.5min) daje linearnu zavisnost s korelacijskim koeficijentom R2 = 0,9985. Pojava drugog signala mogla bi biti posljedica interakcije doksorubicina sa staničnim komponentama u krvi u prisutnosti AgNO3 te nastajanja kompleksa smanjene polarnosti. Površina ispod signala na kromatogramu određena na temelju fluorescencijske detekcije na 480 nm i 560 nm (valne duljine pobuđivanja, odnosno emisije) upotrebljena je za kvantitativnu analizu doksorubicina. Određivanje doksorubicina u krvi psa je jednostavna, precizna i točna. U svim eksperimentima, relativna standardna devijacija (RSD) najčešće je bila < 10%, a izmjerena i teorijska koncentracija razlikovala se za < 10%. Doksorubicin inkubiran s uzorcima krvi psa bio je stabilan najmanje 3 dana ako je čuvan na 5 °C, odnosno najmanje 8 h ako je čuvan na sobnoj temperaturi (28 °C). Predložena HPLC metoda mogla bi imati prednosti u određivanju doksorubicina u sustavima za isporuku lijekova ciljano u krvne stanice i kvantifikaciji ukupne koncentracije doksorubicina u krvi

Brza i osjetljiva HPLC metoda za određivanje doksorubicina u krvi psa Artifakt srebrovog nitrata

A rapid and sensitive high performance liquid chromatographic (HPLC) assay utilizing fluorimetric detection (excitation at 480 nm, emission at 560 nm) for the determination of doxorubicin in dog blood was developed and validated. Treatment of blood samples containing doxorubicin with AgNO3 (as protein precipitant) resulted in appearance of an additional peak in the chromatogram of doxorubicin at 11.5 min along with the parent peak (tR = 5.5 min). The latter peak was not found when treated with other protein precipitants such as trichloroacetic acid and methanol. Construction of calibration curve based on area of both peaks alone did not result in linearity of the curve. However, summation of areas of both peaks resulted in a curve with good linearity and correlation coefficient (R2 = 0.9985). Appearance of second peak may be due to the interaction of doxorubicin with cellular components of blood in the presence of AgNO3 leading to the formation of complex with reduced polarity. Analysis of the quality control samples showed good accuracy (96.7-100.42) and precision (RSD = 2,6-5,7%). The proposed method could be advantageous in estimation of doxorubicin incorporated into targeted delivery systems that concentrate in blood cells and quantify the absolute blood concentration of doxorubicin.Razvijena je i validirana brza i osjetljiva metoda visokotlačne tekućinske kromatografije (HPLC) s fluorimetrijskom detekcijom za određivanje doksorubicina u krvi psa. Nakon obrade uzoraka krvi koji sadrže doksorubicin s AgNO3 (taložno sredstvo za proteine), uz osnovni signal (tR = 5,5min) pojavljuje se dodatni signal u kromatogramu doksorubicina na 11,5 min. Nikakav dodatni signal se ne pojavljuje ako se za taloženje proteina upotrijebe trikloroctena kiselina ili metanol. Međutim, u hemoliziranim uzorcima plazme kojima je dodan doksorubicin nakon obrade sa srebrovim nitratom pojavljuje se drugi signal. Kalibracijska krivulja s površinom ispod signala za samo prvi pik (tR = 5,5min) nije linearna. Međutim, suma površina ispod prvog (tR = 5.5min) i drugog signala (tR = 11.5min) daje linearnu zavisnost s korelacijskim koeficijentom R2 = 0,9985. Pojava drugog signala mogla bi biti posljedica interakcije doksorubicina sa staničnim komponentama u krvi u prisutnosti AgNO3 te nastajanja kompleksa smanjene polarnosti. Površina ispod signala na kromatogramu određena na temelju fluorescencijske detekcije na 480 nm i 560 nm (valne duljine pobuđivanja, odnosno emisije) upotrebljena je za kvantitativnu analizu doksorubicina. Određivanje doksorubicina u krvi psa je jednostavna, precizna i točna. U svim eksperimentima, relativna standardna devijacija (RSD) najčešće je bila < 10%, a izmjerena i teorijska koncentracija razlikovala se za < 10%. Doksorubicin inkubiran s uzorcima krvi psa bio je stabilan najmanje 3 dana ako je čuvan na 5 °C, odnosno najmanje 8 h ako je čuvan na sobnoj temperaturi (28 °C). Predložena HPLC metoda mogla bi imati prednosti u određivanju doksorubicina u sustavima za isporuku lijekova ciljano u krvne stanice i kvantifikaciji ukupne koncentracije doksorubicina u krvi

Endo-Epicardial Homogenization of the Scar Versus Limited Substrate Ablation for the Treatment of Electrical Storms in Patients With Ischemic Cardiomyopathy

ObjectivesThis study investigated the impact on recurrences of 2 different substrate approaches for the treatment of these arrhythmias.BackgroundCatheter ablation of electrical storms (ES) for ventricular arrhythmias (VAs) has shown moderate long-term efficacy in patients with ischemic cardiomyopathy.MethodsNinety-two consecutive patients (81% male, age 62 ± 13 years) with ischemic cardiomyopathy and ES underwent catheter ablation. Patients were treated either by confining the radiofrequency lesions to the endocardial surface with limited substrate ablation (Group 1, n = 49) or underwent endocardial and epicardial ablation of abnormal potentials within the scar (homogenization of the scar, Group 2, n = 43). Epicardial access was obtained in all Group 2 patients, whereas epicardial ablation was performed in 33% (14) of these patients.ResultsMean ejection fraction was 27 ± 5. During a mean follow-up of 25 ± 10 months, the VAs recurrence rate of any ventricular tachycardia (VTs) was 47% (23 of 49 patients) in Group 1 and 19% (8 of 43 patients) in Group 2 (log-rank p = 0.006). One patient in Group 1 and 1 patient in Group 2 died at follow-up for noncardiac reasons.ConclusionsOur study demonstrates that ablation using endo-epicardial homogenization of the scar significantly increases freedom from VAs in ischemic cardiomyopathy patients

Discrepant comorbidity between minority and white suicides: a national multiple cause-of-death analysis

Abstract Background Clinician training deficits and a low and declining autopsy rate adversely impact the quality of death certificates in the United States. Self-report and records data for the general population indicate that proximate mental and physical health of minority suicides was at least as poor as that of white suicides. Methods This cross-sectional mortality study uses data from Multiple Cause-of-Death (MCOD) public use files for 1999–2003 to describe and evaluate comorbidity among black, Hispanic, and white suicides. Unintentional injury decedents are the referent for multivariate analyses. Results One or more mentions of comorbid psychopathology are documented on the death certificates of 8% of white male suicides compared to 4% and 3% of black and Hispanic counterparts, respectively. Corresponding female figures are 10%, 8%, and 6%. Racial-ethnic discrepancies in the prevalence of comorbid physical disease are more attenuated. Cross-validation with National Violent Death Reporting System data reveals high relative underenumeration of comorbid depression/mood disorders and high relative overenumeration of schizophrenia on the death certificates of both minorities. In all three racial-ethnic groups, suicide is positively associated with depression/mood disorders [whites: adjusted odds ratio (AOR) = 31.9, 95% CI = 29.80–34.13; blacks: AOR = 60.9, 95% CI = 42.80–86.63; Hispanics: AOR = 34.7, 95% CI = 23.36–51.62] and schizophrenia [whites: AOR = 2.4, 95% CI = 2.07–2.86; blacks: AOR = 4.2, 95% CI = 2.73–6.37; Hispanics: AOR = 4.1, 95% CI = 2.01–8.22]. Suicide is positively associated with cancer in whites [AOR = 1.8, 95% CI = 1.69–1.93] and blacks [AOR = 1.8, 95% CI = 1.36–2.48], but not with HIV or alcohol and other substance use disorders in any group under review. Conclusion The multivariate analyses indicate high consistency in predicting suicide-associated comorbidities across racial-ethnic groups using MCOD data. However, low prevalence of documented comorbid psychopathology in suicides, and concomitant racial-ethnic discrepancies underscore the need for training in death certification, and routinization and standardization of timely psychological autopsies in all cases of suicide, suspected suicide, and other traumatic deaths of equivocal cause

Ablation of Stable VTs Versus Substrate Ablation in Ischemic Cardiomyopathy the VISTA Randomized Multicenter Trial

Background Catheter ablation reduces ventricular tachycardia (VT) recurrence and implantable cardioverter defibrillator shocks in patients with VT and ischemic cardiomyopathy. The most effective catheter ablation technique is unknown. Objectives This study determined rates of VT recurrence in patients undergoing ablation limited to clinical VT along with mappable VTs ("clinical ablation") versus substrate-based ablation. Methods Subjects with ischemic cardiomyopathy and hemodynamically tolerated VT were randomized to clinical ablation (n = 60) versus substrate-based ablation that targeted all "abnormal" electrograms in the scar (n = 58). Primary endpoint was recurrence of VT. Secondary endpoints included periprocedural complications, 12-month mortality, and rehospitalizations. Results At 12-month follow-up, 9 (15.5%) and 29 (48.3%) patients had VT recurrence in substrate-based and clinical VT ablation groups, respectively (log-rank p < 0.001). More patients undergoing clinical VT ablation (58%) were on antiarrhythmic drugs after ablation versus substrate-based ablation (12%; p < 0.001). Seven (12%) patients with substrate ablation and 19 (32%) with clinical ablation required rehospitalization (p = 0.014). Overall 12-month mortality was 11.9%; 8.6% in substrate ablation and 15.0% in clinical ablation groups, respectively (log-rank p = 0.21). Combined incidence of rehospitalization and mortality was significantly lower with substrate ablation (p = 0.003). Periprocedural complications were similar in both groups (p = 0.61). Conclusions An extensive substrate-based ablation approach is superior to ablation targeting only clinical and stable VTs in patients with ischemic cardiomyopathy presenting with tolerated VT

Pigeonpea improvement: An amalgam of breeding and genomic research

In the past five decades, constant research has been directed towards yield improvement in pigeonpea resulting in the deployment of several commercially acceptable cultivars in India. Though, the genesis of hybrid technology, the biggest breakthrough, enigma of stagnant productivity still remains unsolved. To sort this productivity disparity, genomic research along with conventional breeding was successfully initiated at ICRISAT. It endowed ample genomic resource providing insight in the pigeonpea genome combating production constraints in a precise and speedy manner. The availability of the draft genome sequence with a large‐scale marker resource, oriented the research towards trait mapping for flowering time, determinacy, fertility restoration, yield attributing traits and photo‐insensitivity. Defined core and mini‐core collection, still eased the pigeonpea breeding being accessible for existing genetic diversity and developing stress resistance. Modern genomic tools like next‐generation sequencing, genome‐wide selection helping in the appraisal of selection efficiency is leading towards next‐generation breeding, an awaited milestone in pigeonpea genetic enhancement. This paper emphasizes the ongoing genetic improvement in pigeonpea with an amalgam of conventional breeding as well as genomic research

Modeling causes of death: an integrated approach using CODEm

Background: Data on causes of death by age and sex are a critical input into health decision-making. Priority setting in public health should be informed not only by the current magnitude of health problems but by trends in them. However, cause of death data are often not available or are subject to substantial problems of comparability. We propose five general principles for cause of death model development, validation, and reporting.Methods: We detail a specific implementation of these principles that is embodied in an analytical tool - the Cause of Death Ensemble model (CODEm) - which explores a large variety of possible models to estimate trends in causes of death. Possible models are identified using a covariate selection algorithm that yields many plausible combinations of covariates, which are then run through four model classes. The model classes include mixed effects linear models and spatial-temporal Gaussian Process Regression models for cause fractions and death rates. All models for each cause of death are then assessed using out-of-sample predictive validity and combined into an ensemble with optimal out-of-sample predictive performance.Results: Ensemble models for cause of death estimation outperform any single component model in tests of root mean square error, frequency of predicting correct temporal trends, and achieving 95% coverage of the prediction interval. We present detailed results for CODEm applied to maternal mortality and summary results for several other causes of death, including cardiovascular disease and several cancers.Conclusions: CODEm produces better estimates of cause of death trends than previous methods and is less susceptible to bias in model specification. We demonstrate the utility of CODEm for the estimation of several major causes of death