Jednostavna i brza metoda određivanja udjela ukupnih karotenoida u liofiliziranoj marelici (Prunus armeniaca L.) pomoću refleksne kolorimetrije i fotoakustične spektroskopije

Photoacoustic spectroscopy (PAS) and reflectance colorimetry are suggested as new tools for the analysis of total carotenoids in lyophilized apricot powders. The data obtained by these two techniques from seven apricot cultivars were compared to those acquired by spectrophotometry and high-performance liquid chromatography (HPLC). Best correlations were found between the total carotenoid (TC) content (obtained by VIS spectrophotometry: 1.2–3.4 mg per 100 g of fresh mass) and colorimetric index a* (a* represents the redness of the investigated sample), as well as either argon-ion laser- or xenon-lamp-based PAS. In all three cases linear correlations were comparable. However, according to the sensitivity and precision data, expressed via limit of detection (LOD) and measurement repeatability, the Xe-lamp-based PAS is a preferred approach, followed by colorimetric index a* and Ar-ion laser PAS. Both PAS methods exhibit practically the same Pearson\u27s correlation coefficient (R=0.987 and R=0.991) values. Nevertheless, residual sum of squares (RSS) and residual standard deviation of the linear regression (sy/x) differ markedly. For Xe-lamp-based PAS these parameters were much lower than in the case of Ar-ion laser PAS. Likewise, analysis imprecision amounted to relative standard deviation (RSD) of 1–3 % for Xe-lamp PAS and 2–6 % for Ar-ion laser PAS. On the other hand, as expected, the calibration sensitivity achieved for the PAS signal induced by an Ar-ion laser at 481 nm was substantially higher than that of a Xe-lamp at 470 nm. Nevertheless, according to much lower sy/x, the corresponding LOD for Xe-lamp PAS was still two times lower than that of Ar-ion-based laser PAS (0.59 vs. 1.10 mg per 100 g). Unlike this, Ar-ion laser PAS showed more favourable instrumental precision and standard error of the weighed mean when compared to the Xe-lamp PAS (0.1–0.6 and 0.1–0.3 % vs. 0.5–8.0 and 0.4–1.7 %, respectively). As far as colorimetric indices are concerned, only a* proved to be analytically useful; excellent R but rather modest RSS and sy/x resulted in LOD value of 0.70 mg per 100 g and acceptable analysis imprecision of up to 3 %. The outcome of this research provides sufficient amount of evidence that analytical methods such as reflectance colorimetry and PAS without the use of any chemicals are feasible for reliable quantification of total carotenoids in freeze-dried apricot homogenates.Fotoakustična spektroskopija (PAS) i refleksna kolorimetrija predložene su kao nove metode određivanja udjela ukupnih karotenoida u liofiliziranom prahu marelice. Rezultati dobiveni ispitivanjem sedam kultivara marelice ovim dvjema tehnikama uspoređeni su s onima dobivenim konvencionalnom vidljivom spektrofotometrijom i tekućinskom kromatografijom visoke djelotvornosti (HPLC). Najbolje su korelacije dobivene između udjela ukupnih karotenoida (vidljivom je spektrofotometrijom utvrđen udjel od 1,2 do 3,4 mg u 100 g svježe mase) i kolorimetrijskog indeksa a* (parametar crvenila ispitanog uzorka), te metodom PAS s argonskim laserom ili ksenonskom lampom kao izvorom zračenja. U sva su tri slučaja dobivene usporedljive linearne korelacije. Međutim, s obzirom na osjetljivost i preciznost rezultata izraženih pomoću granice detekcije i ponovljivosti mjerenja, najboljom se pokazala metoda PAS sa ksenonskom lampom, zatim kolorimetrijski indeks a* i naposljetku metoda PAS s argonskim laserom. Obje su metode PAS imale podjednake Pearsonove koeficijente korelacije (R=0,987 i R=0,991). Ipak, rezidualna suma kvadrata (RSS) i rezidualna standardna devijacija linijske regresije (sy/x) značajno su se razlikovale. Za metodu PAS sa ksenonskom lampom ovi su parametri bili znatno manji nego za metodu PAS s argonskim laserom. Također, nepreciznost je metode izražena pomoću relativne standardne devijacije (RSD) bila od 1 do 3 % za metodu PAS sa ksenonskom lampom i od 2 do 6 % za metodu PAS s argonskim laserom. S druge je pak strane, kao što je i bilo očekivano, kalibracijska osjetljivost postignuta za signal PAS induciran argonskim laserom pri 481 nm bila znatno niža od one dobivene ksenonskom lampom pri 470 nm. Ipak, zahvaljujući bitno nižoj vrijednosti sy/x, granica detekcije metode PAS sa ksenonskom lampom (0,59 mg u 100 g) bila je dvostruko manja od one metode PAS s argonskim laserom (1,10 mg u 100 g). Nasuprot tomu, metoda PAS s argonskim laserom imala je bolju preciznost instrumenta (nepreciznost mjerenja od samo 0,1-0,6 %) i standardnu pogrešku ponderirane sredine (0,1-0,3 %), u usporedbi s metodom PAS sa ksenonskom lampom (0,5-8,0 % odnosno 0,4-1,7 %). Među kolorimetrijskim indeksima samo je indeks a* bio koristan za analizu; odlične vrijednosti R, ali skromne vrijednosti RSS i sy/x rezultirale su granicom detekcije od 0,70 mg u 100 g i prihvatljivom nepreciznošću mjerenja od najviše 3 %. Rezultati ovog istraživanja pokazuju da se analitičke tehnike, poput refleksne kolorimetrije i fotoakustične spektroskopije mogu primijeniti za pouzdano određivanje udjela ukupnih karotenoida u liofiliziranim homogenatima marelice, i to bez uporabe reagenasa

Jednostavna i brza metoda određivanja udjela ukupnih karotenoida u liofiliziranoj marelici (Prunus armeniaca L.) pomoću refleksne kolorimetrije i fotoakustične spektroskopije

Photoacoustic spectroscopy (PAS) and reflectance colorimetry are suggested as new tools for the analysis of total carotenoids in lyophilized apricot powders. The data obtained by these two techniques from seven apricot cultivars were compared to those acquired by spectrophotometry and high-performance liquid chromatography (HPLC). Best correlations were found between the total carotenoid (TC) content (obtained by VIS spectrophotometry: 1.2–3.4 mg per 100 g of fresh mass) and colorimetric index a* (a* represents the redness of the investigated sample), as well as either argon-ion laser- or xenon-lamp-based PAS. In all three cases linear correlations were comparable. However, according to the sensitivity and precision data, expressed via limit of detection (LOD) and measurement repeatability, the Xe-lamp-based PAS is a preferred approach, followed by colorimetric index a* and Ar-ion laser PAS. Both PAS methods exhibit practically the same Pearson\u27s correlation coefficient (R=0.987 and R=0.991) values. Nevertheless, residual sum of squares (RSS) and residual standard deviation of the linear regression (sy/x) differ markedly. For Xe-lamp-based PAS these parameters were much lower than in the case of Ar-ion laser PAS. Likewise, analysis imprecision amounted to relative standard deviation (RSD) of 1–3 % for Xe-lamp PAS and 2–6 % for Ar-ion laser PAS. On the other hand, as expected, the calibration sensitivity achieved for the PAS signal induced by an Ar-ion laser at 481 nm was substantially higher than that of a Xe-lamp at 470 nm. Nevertheless, according to much lower sy/x, the corresponding LOD for Xe-lamp PAS was still two times lower than that of Ar-ion-based laser PAS (0.59 vs. 1.10 mg per 100 g). Unlike this, Ar-ion laser PAS showed more favourable instrumental precision and standard error of the weighed mean when compared to the Xe-lamp PAS (0.1–0.6 and 0.1–0.3 % vs. 0.5–8.0 and 0.4–1.7 %, respectively). As far as colorimetric indices are concerned, only a* proved to be analytically useful; excellent R but rather modest RSS and sy/x resulted in LOD value of 0.70 mg per 100 g and acceptable analysis imprecision of up to 3 %. The outcome of this research provides sufficient amount of evidence that analytical methods such as reflectance colorimetry and PAS without the use of any chemicals are feasible for reliable quantification of total carotenoids in freeze-dried apricot homogenates.Fotoakustična spektroskopija (PAS) i refleksna kolorimetrija predložene su kao nove metode određivanja udjela ukupnih karotenoida u liofiliziranom prahu marelice. Rezultati dobiveni ispitivanjem sedam kultivara marelice ovim dvjema tehnikama uspoređeni su s onima dobivenim konvencionalnom vidljivom spektrofotometrijom i tekućinskom kromatografijom visoke djelotvornosti (HPLC). Najbolje su korelacije dobivene između udjela ukupnih karotenoida (vidljivom je spektrofotometrijom utvrđen udjel od 1,2 do 3,4 mg u 100 g svježe mase) i kolorimetrijskog indeksa a* (parametar crvenila ispitanog uzorka), te metodom PAS s argonskim laserom ili ksenonskom lampom kao izvorom zračenja. U sva su tri slučaja dobivene usporedljive linearne korelacije. Međutim, s obzirom na osjetljivost i preciznost rezultata izraženih pomoću granice detekcije i ponovljivosti mjerenja, najboljom se pokazala metoda PAS sa ksenonskom lampom, zatim kolorimetrijski indeks a* i naposljetku metoda PAS s argonskim laserom. Obje su metode PAS imale podjednake Pearsonove koeficijente korelacije (R=0,987 i R=0,991). Ipak, rezidualna suma kvadrata (RSS) i rezidualna standardna devijacija linijske regresije (sy/x) značajno su se razlikovale. Za metodu PAS sa ksenonskom lampom ovi su parametri bili znatno manji nego za metodu PAS s argonskim laserom. Također, nepreciznost je metode izražena pomoću relativne standardne devijacije (RSD) bila od 1 do 3 % za metodu PAS sa ksenonskom lampom i od 2 do 6 % za metodu PAS s argonskim laserom. S druge je pak strane, kao što je i bilo očekivano, kalibracijska osjetljivost postignuta za signal PAS induciran argonskim laserom pri 481 nm bila znatno niža od one dobivene ksenonskom lampom pri 470 nm. Ipak, zahvaljujući bitno nižoj vrijednosti sy/x, granica detekcije metode PAS sa ksenonskom lampom (0,59 mg u 100 g) bila je dvostruko manja od one metode PAS s argonskim laserom (1,10 mg u 100 g). Nasuprot tomu, metoda PAS s argonskim laserom imala je bolju preciznost instrumenta (nepreciznost mjerenja od samo 0,1-0,6 %) i standardnu pogrešku ponderirane sredine (0,1-0,3 %), u usporedbi s metodom PAS sa ksenonskom lampom (0,5-8,0 % odnosno 0,4-1,7 %). Među kolorimetrijskim indeksima samo je indeks a* bio koristan za analizu; odlične vrijednosti R, ali skromne vrijednosti RSS i sy/x rezultirale su granicom detekcije od 0,70 mg u 100 g i prihvatljivom nepreciznošću mjerenja od najviše 3 %. Rezultati ovog istraživanja pokazuju da se analitičke tehnike, poput refleksne kolorimetrije i fotoakustične spektroskopije mogu primijeniti za pouzdano određivanje udjela ukupnih karotenoida u liofiliziranim homogenatima marelice, i to bez uporabe reagenasa

Viruses: incredible nanomachines. New advances with filamentous phages

During recent decades, bacteriophages have been at the cutting edge of new developments in molecular biology, biophysics, and, more recently, bionanotechnology. In particular filamentous viruses, for example bacteriophage M13, have a virion architecture that enables precision building of ordered and defect-free two and three-dimensional structures on a nanometre scale. This could not have been possible without detailed knowledge of coat protein structure and dynamics during the virus reproduction cycle. The results of the spectroscopic studies conducted in our group compellingly demonstrate a critical role of membrane embedment of the protein both during infectious entry of the virus into the host cell and during assembly of the new virion in the host membrane. The protein is effectively embedded in the membrane by a strong C-terminal interfacial anchor, which together with a simple tilt mechanism and a subtle structural adjustment of the extreme end of its N terminus provides favourable thermodynamical association of the protein in the lipid bilayer. This basic physicochemical rule cannot be violated and any new bionanotechnology that will emerge from bacteriophage M13 should take this into account

Assaying Total Carotenoids in Flours of Corn and Sweetpotato by Laser Photoacoustic Spectroscopy

This study describes the application of the laser photoacoustic spectroscopy (PAS) for quantification of total carotenoids (TC) in corn flours and sweetpotato flours. Overall, thirty-three different corn flours and nine sweetpotato flours were investigated. All PAS measurements were performed at room temperature using 488-nm argon laser radiation for excitation and mechanical modulation of 9 and 30 Hz. The measurements were repeated within a run and within several days or months. The UV–Vis spectrophotometry was used as the reference method. The concentration range that allows for the reliable analysis of TC spans a region from 1 to 40 mg kg−1 for corn flours and from 9 to 40 mg kg−1 for sweetpotato flours. In the case of sweetpotato flours, the quantification may extend even to 240 mg kg−1 TC. The estimated detection limit values for TC in corn and sweetpotato flours were 0.1 and 0.3 mg kg−1, respectively. The computed repeatability (n = 3–12) and intermediate precision (n = 6–28) RSD values at 9 and 30 Hz are comparable: 0.1–17.1% and 5.3–14.7% for corn flours as compared with 1.4–9.1% and 4.2–23.0% for sweetpotato flours. Our results show that PAS can be successfully used as a new analytical tool to simply and rapidly screen the flours for their nutritional potential based on the total carotenoid concentration

Unraveling genetic predisposition to familial or early onset gastric cancer using germline whole-exome sequencing

Recognition of individuals with a genetic predisposition to gastric cancer (GC) enables preventive measures. However, the underlying cause of genetic susceptibility to gastric cancer remains largely unexplained. We performed germline whole-exome sequencing on leukocyte DNA of 54 patients from 53 families with genetically unexplained diffuse-type and intestinal-type GC to identify novel GC-predisposing candidate genes. As young age at diagnosis and familial clustering are hallmarks of genetic tumor susceptibility, we selected patients that were diagnosed below the age of 35, patients from families with two cases of GC at or below age 60 and patients from families with three GC cases at or below age 70. All included individuals were tested negative for germline CDH1 mutations before or during the study. Variants that were possibly deleterious according to in silico predictions were filtered using several independent approaches that were based on gene function and gene mutation burden in controls. Despite a rigorous search, no obvious candidate GC predisposition genes were identified. This negative result stresses the importance of future research studies in large, homogeneous cohorts