11 research outputs found
Primena LCāMS/MS metoda u ispitivanju parametara oksidativnog stresa
Oxidative stress is a phenomenon that occurs due to the disturbance in the balance
between the production of reactive oxygen species and the ability of biological systems to
remove the resulting compounds. Oxidative stress is involved in the pathogenesis of many
disorders in the human organism. This indicates the importance of quantification of
oxidative stress parameters in biological samples. Traditionally, these parameters are
determined by biochemical tests. Although these tests are routinely performed, they have
many drawbacks. To determine the exact concentration of selected compounds, more
sensitive analytical methods are becoming more important. In the modern scientific
literature, liquid chromatography-tandem mass spectrometry (LC-MS/MS) is increasingly
mentioned (1). This method, with adequate selection of stationary and mobile phases,
enables quantification of very low concentrations of selected parameters. In addition, these
methods can simultaneously determine the concentration of many selected components.
However, it is necessary to take into account that LC-MS/MS methods require a very purified
biological sample from which the proteins have been maximally removed. In this work, we
will present the results of determination of cysteine, cystine, reduced and oxidized
glutathione in patients with microcellular lung cancer. The use of LC-MS/MS methods is
becoming increasingly common for the analysis of oxidative stress markers in biological
fluids. In the future, we are expected to move to even more modern, fully automated
methods, which simultaneously purify and analyze samples.Oksidativni stres je pojava koja nastaje usled naruÅ”avanja ravnoteže izmeÄu
proizvodnje i nagomilavanja reaktivnih jedinjenja kiseonika u organizmu i sposobnosti
bioloÅ”kih sistema da ukloni nastala jedinjenja. Oksidativni stres uÄestvuje u patogenezi
mnogih poremeÄaja kao Å”to su kardiovaskularne bolesti, dijabetes i bolesti bubrega. Ovo
ukazuje na znaÄaj odreÄivanja odabranih parametara oksidativnog stresa u uzorcima
bioloÅ”kog materijala. Tradicionalno, najveÄi broj parametara se odreÄuje biohemijskim
testovima. Iako se ovi testovi rutinski izvode, oni imaju mnoge nedostatke. Da bi se odredila
taÄna koncentracija izabranih parametara, koji se u uzorcima nalaze u veoma niskim
koncentracijama, sve veÄu prednost imaju osetljivije analitiÄke metode. U savremenoj
nauÄnoj literaturi se u te svrhe sve viÅ”e spominje teÄna hromatografija spregnuta sa
masenim detektorom (LC-MS/MS) (1). Ova metoda, uz adekvatan izbor stacionarne i
mobilne faze, omoguÄava kvantifikaciju veoma niskih koncentracija odabranih jedinjenja.
Sem toga, ovim metodama se istovremeno, u jednom uzorku, može odrediti koncentracija
veÄeg broja odabranih komponenti. Ipak, neophodno je voditi raÄuna o tome da teÄna
hromatografija spregnuta sa masenim detektorom zahteva dobro preÄiÅ”Äen uzorak bioloÅ”kog
materijala iz kog su maksimalno uklonjeni proteini. U ovom radu biÄe prikazani rezultati
odreÄivanja cisteina, cistina, redukovanog i oksidovanog glutationa kod pacijenata sa
mikrocelularnim karcinomom pluÄa. Upotreba LC-MS/MS metoda je sve uobiÄajenija za
analizu markera oksidativnog stresa u bioloÅ”kim teÄnostima zbog svoje osetljivosti. U
buduÄnosti se oÄekuje razvoj ka joÅ” savremenijim, potpuno automatizovanim metodama,
kojima se istovremeno uzorci preÄiÅ”Äavaju i analiziraju.VIII Kongres farmaceuta Srbije sa meÄunarodnim uÄeÅ”Äem, 12-15.10.2022. Beogra
Analysis of the Prognostic Potential of Schlafen 11, Programmed Death Ligand 1, and Redox Status in Colorectal Cancer Patients
The Schlafen 11 (SLFN11) protein has recently emerged as pivotal in DNA damage conditions, with predictive potential for tumor response to cytotoxic chemotherapies. Recent discoveries also showed that the programmed death ligand 1 (PD-L1) protein can be found on malignant cells, providing an immune evasion mechanism exploited by different tumors. Additionally, excessive generation of free radicals, redox imbalance, and consequential DNA damage can affect intestinal cell homeostasis and lead to neoplastic transformation. Therefore, our study aimed to investigate the significance of SLFN11 and PD-L1 proteins and redox status parameters as prognostic biomarkers in CRC patients. This study included a total of 155 CRC patients. SLFN11 and PD-L1 serum levels were measured with ELISA and evaluated based on redox status parameters, sociodemographic and clinical characteristics, and survival. The following redox status parameters were investigated: spectrophotometrically measured superoxide dismutase (SOD), sulfhydryl (SH) groups, advanced oxidation protein products (AOPP), malondialdehyde (MDA), pro-oxidantāantioxidant balance (PAB), and superoxide anion (O2ā¢ā). The prooxidative score, antioxidative score, and OXY-SCORE were also calculated. The results showed significantly shorter survival in patients with higher OXY-SCOREs and higher levels of serum SLFN11, while only histopathology-analysis-related factors showed significant prognostic value. OXY-SCORE and SLFN11 levels may harbor prognostic potential in CRC patients
Oksidativni stres i dužina telomera kod COVIDā19 pacijenata
The pathogenesis of the COVID-19 infection caused by the SARS-CoV-2 coronavirus is
the subject of active research worldwide. As a complex disease, it is suspected that oxidative
stress plays a role in its pathogenesis and contributes to the progressive shortening of
telomeres. Patients aged 25ā84 years (n=31) were monitored at the time of diagnosis, as
well as at control on the 14th and 21st days. Redox status parameters were determined: total
oxidant status (TOS), superoxide anion radical (O2.-), prooxidant-antioxidant balance (PAB),
advanced protein oxidation products (AOPP), malondialdehyde (MDA), and ischemia-
modified albumin (IMA) as prooxidants and total antioxidant status (TAS), superoxide-
dismutase (SOD) and paraoxonase 1 (PON1) enzyme activity, total antioxidant and total
oxidant status ratio (TAS/TOS), and total sulfhydryl groups (SHG) as antioxidants. Patients
have the shortest telomeres at diagnosis and their length increases during recovery. A
statistically significant difference in rTL was shown in patients at the time of diagnosis and
after 21 days (p=0.008). rTL has a statistically significant negative correlation with SHG in
patients at the time of diagnosis (Ļ=-0.386, p=0.038), as well as in patients after 14 days (Ļ =-
0.389, p=0.037). After 21 days, a statistically significant positive correlation was shown
between rTL and SOD (Ļ =0.381, p=0.046). The obtained results indicate progressive
shortening of telomeres caused by COVID-19 infection, impaired redox status, as well as the
change of these parameters during recovery, which confirms the link between oxidative
stress and rTL, as well as the participation of these processes in the pathogenesis of COVID-
19.Patogeneza COVID-19 infekcije izazvane koronavirusom SARS-CoV-2 je predmet
aktivnog istraživanja Ŕirom sveta. Kako je u pitanju složena bolest, smatra se da i u njenoj
patogenezi leži oksidativni stres, koji doprinosi progesivnom skraÄivanju telomera. Pacijenti
starosti 25-84 godina (n=31) praÄeni su u trenutku postavljanja dijagnoze, kao i na kontroli
14. i 21. dana. Parametri redoks statusa odreÄivani su iz seruma ili plazme odgovarajuÄim
spektrofotometrijskim metodama, dok je iz leukocita periferne krvi merena relativna dužina
telomera (rTL) pomoÄu qPCR metode. Od parametara oksidativnog statusa odreÄivani su:
totalni oksidantni status (TOS), superoksidni anjon radikal (O2.- ), prooksidantno-
antioksidantni balans (PAB), uznapredovali produkti oksidacije proteina (AOPP),
malondialdehid (MDA), ishemijom modifikovan albumin (IMA). Kao parametri
antioksidativne zaÅ”tite odreÄivani su: totalni antioksidantni status (TAS), aktivnost enzima
superoksid-dismutaze (SOD) i paraoksonaze 1 (PON1), odnos totalnog antioksidantnog i
totalnog oksidantnog statusa (TAS/TOS), kao i ukupne sulfhidrilne grupe (SHG). Pacijenti u
trenutku postavljanja dijagnoze imaju najkraÄe telomere i njihova dužina se poveÄava tokom
oporavka. Pokazana je statistiÄki znaÄajna razlika u rTL kod pacijenata u trenutku
postavljanja dijagnoze i nakon 21 dan (p=0,008). rTL je u statistiÄki znaÄajnoj negativnoj
korelaciji sa SHG kod pacijenata u trenutku postavljanja dijagnoze (Ļ=-0,386, p=0,038), kao i
kod pacijenata nakon 14 dana oporavka (Ļ=-0,389, p=0,037). Nakon 21 dan, pokazana je
statistiÄki znaÄajna pozitivna korelacija izmeÄu rTL i SOD (Ļ=0,381, p=0,046). Dobijeni
rezultati ukazuju na progresivno skraÄenje telomera izazvano COVID-19 infekcijom, naruÅ”en
redoks status, kao i na promenu ovih parametara tokom oporavka, koja potvrÄuje vezu
izmeÄu oksidativnog stresa i rTL, kao i uÄeÅ”Äe ovih procesa u patogenezi COVID-19.VIII Kongres farmaceuta Srbije sa meÄunarodnim uÄeÅ”Äem, 12-15.10.2022. Beogra
Oxidative Stress and Inflammatory Biomarkers in Patients with Diabetic Foot
Background and Objectives: Diabetic foot (DF) development is driven by complex interactions of hyperglycemia, inflammation, and oxidative stress (OS). We aimed to investigate OS and inflammatory biomarkers in patients with DF and their potential to improve early diagnosis and management of DF. Materials and Methods: The prooxidantāantioxidant balance (PAB), superoxide dismutase (SOD), total oxidative status (TOS), total sulfhydryl groups (SHG), routine biochemical parameters, and complete blood count were determined in 42 patients with type-2 DM, of which 23 patients had DF, while 19 patients were without DF complications. The neutrophils-to-lymphocyte ratio (NLR) was evaluated as a biomarker of inflammation. Results: Patients with DF had significantly higher (p < 0.05) PAB levels (170 Ā± 33.9 U/L) compared to those without DF complications (142 Ā± 31.3 U/L). In addition, patients with DF had significantly reduced SOD activities (p < 0.01). NLR values were significantly higher in the DF group (median: 2.8; interquartile range: 2.0ā4.3) than in the group without DF (median: 1.4; interquartile range: 1.4ā2.1; p < 0.01). A positive correlation was found between the PAB and NLR index (r = 0.449; p < 0.05). The diagnostic accuracy of both PAB (AUC = 0.741; p < 0.01) and NLR (AUC = 0.760; p < 0.01) was estimated as acceptable. Conclusions: In conclusion, the development of DF is associated with enhanced OS and inflammation processes. PAB and NLR could be useful non-invasive biomarkers of DF development
Markeri redoks statusa kod pacijenata sa nealkoholnom masnom boleÅ”Äu jetre
Nonalcoholic fatty liver disease (NAFLD) is the most common chronic liver disease,
present in up to 30% of the adult population worldwide (1). Triglycerides accumulation in
hepatocytes (steatosis) represents the root cause of NAFLD and is associated with oxidative
stress, which could further lead to fibrosis and cell death of hepatocytes (2). The aim of this
research was to identify redox status markers for predicting the risk of developing steatosis.
158 participants were included. Steatosis was confirmed by ultrasound in 101 subjects,
while the remaining 57 were in the control group. The following markers of redox status
were determined in serum and plasma samples of all subjects: superoxide dismutase 1
(SOD1), paraoxonase (PON1), malondialdehyde (MDA) and superoxide anion (O2.-). For this
purpose, spectrophotometric methods and enzyme immunosorbent assays were used. SOD1
was statistically significantly higher (P<0.001), while O2 .- was significantly lower in the
patient group (P<0.001). SOD1 was significantly negatively correlated with O2.- (Ļ= -0.494,
P<0.001) and MDA (Ļ= -0.242, P=0.002). Univariate binary logistic regression analysis
showed a positive association between SOD1 and the presence of steatosis (OR=1.018, 95%
CI 1.005-1.031; P=0.005), as well as a negative association between O2 .- and the presence of
steatosis (OR=0.959, 95% CI 0.941-0.978; P<0.001). Multivariate analysis singled out SOD1
(OR=1.024, 95% CI 1.006-1.041; P=0.007) and O2.- (OR=0.965, 95% CI 0.942-0.989;
P=0.004) as independent predictors for the presence of steatosis in our subjects. The redox
status parameters, SOD1 and O2, respectively, showed a positive and negative prediction of
the presence of statosis in our subjects.Nealkoholna masna bolest jetre (eng. nonalcoholic fatty liver disease, NAFLD) je
najÄeÅ”Äe hroniÄno oboljenje jetre, prisutno i u do 30% adultne populacije Å”irom sveta (1).
ZnaÄajnu ulogu u nastanku NAFLD ima akumulacija triglicerida u hepatocitima ā steatoza,
koja je povezana sa oksidativnim stresom, a koji dalje vodi fibrozi i Äelijskoj smrti hepatocita
(2). Cilj ovog istraživanja bio je identifikacija markera redoks statusa za predviÄanje rizika za
nastanak steatoze. Studija je obuhvatila 158 ispitanika iz KliniÄko bolniÄkog centra Zemun.
Steatoza je potvrÄena ultrazvukom kod 101 ispitanika, dok je preostalih 57 Äinilo kontrolnu
grupu. U uzorcima seruma i plazme svih ispitanika odreÄeni su sledeÄi markeri redoks
statusa: superoksid dismutaza 1 (SOD1), paraoksonaza (PON1), malondialdehid (MDA) i
superoksidni anjon (O2.-). U tu svrhu koriÅ”Äene su spektrofotometrijske metode i enzimski
imunosorbentni testovi. SOD1 je bila statistiÄki znaÄajno viÅ”a (P<0,001), dok O 2.- znaÄajno
niži u grupi pacijenata (P<0,001), dok se PON1 i MDA nisu znaÄajno razlikovali izmeÄu
grupa. SOD1 je bila u znaÄajnoj negativnoj korelaciji sa O 2.- (Ļ=-0,494, P<0,001) i MDA (Ļ= -
0,242, P=0,002). Univarijatna binarna logistiÄka regresiona analiza je pokazala pozitivnu
asocijaciju izmeÄu SOD1 i prisustva steatoze (OR=1,018, 95% CI 1,005-1,031; P=0,005), kao i
negativnu asocijaciju izmeÄu O 2.- i prisustva steatoze (OR=0,959, 95% CI 0,941-0,978;
P<0,001). Multivarijantna analiza je izdvojila SOD1 (OR=1,024, 95% CI 1,006-1,041;
P=0,007) i O2.- (OR=0,965, 95% CI 0,942-0,989; P=0,004) kao nezavisne prediktore za
prisustvo steatoze kod naŔih ispitanika. Parametri redoks statusa, SOD1 i O 2.- redom, su
pokazali pozitivnu, odnosno negativnu predikciju prisustva statoze kod naÅ”ih ispitanika.VIII Kongres farmaceuta Srbije sa meÄunarodnim uÄeÅ”Äem, 12-15.10.2022. Beogra
Dužina telomera kao biomarker starenja i bolesti
As research related to healthspan and lifespan has become a hot topic, the necessity for a
reliable and practical biomarker of aging (BoA), which can provide information about mortality
and morbidity risk, along with remaining life expectancy, has increased. The chromosome
terminus non-coding protective structure that prevents genomic instability is called a telomere.
The continual shortening of telomeres, which affects their structure as well as function, is a
hallmark of agedness. The aforementioned process is a potential cause of age-related diseases
(ARDs), leading to a bad prognosis and a low survival rate, which compromise health and
longevity. Hence, studies scrutinizing the BoAs often include telomere length (TL) as a
prospective candidate. The results of these studies suggest that TL measurement can only provide
an approximate appraisal of the aging rate, and its implementation into clinical practice and
routine use as a BoA has many limitations and challenges. Nevertheless, measuring TL while
determining other biomarkers can be used to assess biological age. This review focuses on the
importance of telomeres in health, senescence, and diseases, as well as on summarizing the results
and conclusions of previous studies evaluating TL as a potential BoA.Kako su istraživanja na temu starenja postala sve popularnija, javila se potreba za pouzdanim i praktiÄnim biomarkerom starenja, koji može pružiti informacije o riziku od mortaliteta i morbiditeta. Telomere su nekodirajuÄi krajevi linearnih hromozoma koji održavaju njihovu stabilnost i integritet. Äelijsko starenje i starenje organizma karakteriÅ”e progresivno skraÄivanje telomera, Å”to ugrožava njihovu strukturu i funkciju. SkraÄivanje telomera je u vezi sa poveÄanom incidencom bolesti povezanih sa starenjem i loÅ”om stopom preživljavanja, Å”to ugrožava zdravlje i skraÄuje životni vek. Stoga je dužina telomera dugo vremena prepoznata kao jedan od najboljih biomarkera starenja. MeÄutim, nedavna istraživanja ukazuju na to da dužina telomera može da pruži samo približnu procenu brzine starenja, pa je implementacija ovog biomarkera u kliniÄku praksu i rutinsku primenu praÄena mnogim ograniÄenjima i izazovima. Uprkos tome, merenje dužine telomera, uz istovremeno odreÄivanje drugih biomarkera, može poslužiti za procenu bioloÅ”ke starosti. Fokus ovog rada je na znaÄaju telomera u ljudskom zdravlju, starenju i bolestima, kao i na sumiranju rezultata i zakljuÄaka dosadaÅ”njih studija koje su se bavile ispitivanjem dužine telomera kao potencijalnog kandidata za biomarker starenja
Correlation of Systemic Inflammation Parameters and Serum SLFN11 in Small Cell Lung CancerāA Prospective Pilot Study
Background and objectives: The objective of this research was to analyze the correlation of the neutrophil-to-lymphocyte ratio (NLR), C-reactive protein (CRP), soluble programmed cell death ligand 1 (sPD-L1), and Schlafen 11 (SLFN11) with the response to first-line chemotherapy in a cohort of small cell lung cancer (SCLC) patients, and to determine their potential as predictive serum biomarkers. Materials and Methods: A total of 60 SCLC patients were included. Blood samples were taken to determine CRP, sPD-L1, and SLFN11 levels. The first sampling was performed before the start of chemotherapy, the second after two cycles, and the third after four cycles of chemotherapy. Results: The patients who died earlier during the study had NLR and SLFN11 concentrations significantly higher compared to the survivor group. In the group of survivors, after two cycles of chemotherapy, the NLR ratio decreased significantly (p < 0.01), but after four cycles, the NLR ratio increased (p < 0.05). Their serum SLFN11 concentration increased significantly (p < 0.001) after two cycles of chemotherapy, but after four cycles, the level of SLFN11 fell significantly (p < 0.01). CRP, NLR, and SLFN11 were significant predictors of patient survival according to KaplanāMeier analysis. The combination of inflammatory parameters and SLFN11 with a cutoff value above the 75th percentile of the predicted probability was associated with significantly lower overall survival in SCLC patients (average survival of 3.6 months vs. 4.8 months). Conclusion: The combination of inflammatory markers and the levels of two specific proteins (sPD-L1, SLFN11) could potentially serve as a non-invasive biomarker for predicting responses to DNA-damaging therapeutic agents in SCLC
The influence of Klotho protein and prooxidantāantioxidant balance combination on the mortality of HD patients
Purpose: End-stage renal disease patients on chronic hemodialysis (HD) have a shortened life expectancy compared to the general population. The aim of this study was to evaluate a possible link between three new and emerging factors in renal pathophysiology: Klotho protein, telomere length in peripheral blood mononuclear cells (TL) and redox status parameters before HD (bHD) and after HD (aHD), and to test mortality prediction capability of these emerging parameters in a population of HD patients. Methods: The study included 130 adult patients with average age 66 (54ā72), on HD (3 times per week; 4ā5 h per session). Klotho level, TL, routine laboratory parameters, dialysis adequacy and redox status parameters: advanced oxidation protein products (AOPP), prooxidantāantioxidant balance (PAB), superoxide anion (O2.ā), malondialdehyde (MDA), ischemia-modified albumin (IMA), total sulfhydryl group content (SHG), and superoxide dismutase (SOD) were determined. Results: Klotho concentration was significantly higher aHD; 68.2 (22.6ā152.9) vs. bHD 64.2 (25.5ā119.8) (p = 0.027). The observed increase in TL was not statistically significant. AOPP, PAB, SHG, and SOD activity were significantly increased aHD (p > 0.001). The patients with the highest mortality risk score (MRS) had significantly higher PAB bHD (p = 0.002). Significantly lower O2.ā (p < 0.001), SHG content (p = 0.072), and IMA (p = 0.002) aHD were found in patients with the lowest MRS values. Principal component analysis revealed redox balance-Klotho factor as a significant predictor of high mortality risk (p = 0.014). Conclusion: Decreased Klotho and TL attrition as well as redox status disturbance could be connected with higher mortality rate in HD patients
Childhood obesity accelerates biological ageing: is oxidative stress a link?
Obesity is multifactorial pathophysiological condition with an imbalance in biochemical, immunochemical, redox status and genetic parameters values. We aimed estimate connection between relative leukocyte telomere lengths (rLRL) - biomarker of cellular aging with metabolic and redox status biomarkers values in a group of obese and lean children. The study includes 110 obese and 42 lean children and adolescents, both genders. The results suggested that rLTL are significantly shorter in obese, compared to lean group (p30kg/m2 according to logistic regression analysis was PCA derived antioxidant factor's score (OR: 1.66, 95th Cl: 1.05-2.6, p=0.029). PCA analysis confirmed oxidative stress importance in biological ageing caused by obesity and its multiple consequences related to prooxidants augmentation and antioxidants exhaustion and gave us clear signs of disturbed cellular homeostasis deepness, even before any overt disease occurrence.This is peer-reviewed version of the following article: SuboÅ”iÄ, B., ZdravkoviÄ, V., JeÅ”iÄ, M., Munjas, J., KovaÄeviÄ, S., GuzonjiÄ, A., MitroviÄ, J., Saso, L., ÄuriÄiÄ, I.,& Kotur-StevuljeviÄ, J.. (2024). Childhood obesity accelerates biological ageing: is oxidative stress a link?. in British Journal of Nutrition
Cambridge University Press..
[https://doi.org/10.1017/S0007114524000898
Correlation of Short Leukocyte Telomeres and Oxidative Stress with the Presence and Severity of Lung Cancer Explored by Principal Component Analysis
Lung cancer (LC) is the second most common malignancy and leading cause of cancer death. The potential "culprit" for local and systemic telomere shortening in LC patients is oxidative stress. We investigated the correlation between the peripheral blood leukocyte (PBL) telomere length (TL) and the presence/severity of LC and oxidative stress, and its usefulness as LC diagnostic marker. PBL TL was measured in 89 LC patients and 83 healthy subjects using the modified Cawthon RTq-PCR method. The relative PBL TL, found to be a potential diagnostic marker for LC with very good accuracy (P < 0.001), was significantly shorter in patients compared to the control group (CG) (P < 0.001). Significantly shorter telomeres were found in patients with LC TNM stage IV than in patients with stages I-III (P = 0.014), in patients without therapy compared to those on therapy (P = 0.008), and in patients with partial response and stable/progressive disease compared to those with complete response (P = 0.039). The total oxidant status (TOS), advanced oxidation protein products (AOPP), prooxidant-antioxidant balance (PAB) and C-reactive protein (CRP) were significantly higher in patients compared to CG (P < 0.001) and correlated negatively with TL in both patients and CG (P < 0.001). PCA showed a relation between PAB and TL, and between the EGFR status and TL. Oxidative stress and PBL telomere shortening are probably associated with LC development and progression