Korelacija između proteinske i genske ekspresije HER-2 i topoizomeraze 2a kod infiltrativnog duktalnog karcinoma dojke

Introduction: Breast cancer is a heterogeneous disease and the most common malignancy in women with increasing incidence in recent years. According to immunohistochemical expression of estrogen receptor (ER-R), progesterone receptor (PR-R) and HER-2 four different molecular subtypes of breast cancer were identified: luminal A (ER-R+ and / or PR-R+, HER-2-), luminal B (ER-R+ and / or PR-R+, HER- 2+), HER-2 positive (ER-R- and / or PR-R-, HER-2+) and triple negative (ER-R- and / or PR-R-, HER-2-) breast cancer subtypes. HER-2 and TOP2A genes are amplified in 20– 30 % of breast cancers. Amplification of HER-2 and TOP2A genes are associated with a poor prognosis and predictive response to treatment by Herceptin and anthracycline. Co-amplification of HER-2 and TOP2A has been seen in approximately 12–38 % of patients with breast cancers. A few retrospective studies have reported that TOP2A gene amplification occurs almost exclusively in conjunction with HER-2 gene amplification. Aim: The aim of our study is to investigate HER-2 and TOP2A gene aberrations and their correlation with apoptotic and proliferative indexes, Bcl-2, p53, ER-R, PR-R, HER-2, TOP2A, BRCA1, clinicopathological parameters and overall survival in four molecular subtypes of breast cancer. Material and methods: Sixty paraffin-embedded breast cancer tissue samples were studied immunohistochemically for the expression of ER-R, PR-R, HER-2, p53, Bcl-2, ssDNA, Ki67, BRCA1, TOP2A and FISH for HER-2 and TOP2A gene aberrations. HER-2 and TOP2A gene copy number were analyzed by HER-2 FISH pharmDxTM kit and TOP2A FISH pharmDxTM kit (DakoCytomation), respectively. Results: Clinicopathological parameters - The ages of patients ranged from 38 to 87 years (64.79 ± 21). The tumor size ranged between 0.4 and 5.5 cm, and the histological grades were G1, G2 and G3 in 6 (10 %), 25 (41.7 %), and 17 (28.3 %) patients, respectively. In 26 patients (43.4 %) lymph node status was negative; while in 22 patients (36.6 %) was detected positive lymph node status...Uvod: Karcinom dojke predstavlja jako heterogeno oboljenje kao i jedno od najzastupljenijih malignih bolesti u opštoj ženskoj populaciji. Na osnovu prisustva i nivoa ekspresije ER-R, PR-R i HER-2 receptora ostvarena je podela na četiri molekularna tipa karcinoma dojke: luminalni A (ER-R+ i / ili PR-R+, HER-2-), luminalni B (ER-R+ i / ili PR-R+, HER-2+), HER-2 pozitivni (ER-R- i / ili PR-R-, HER-2+) i trostruko negativni (ER-R- i / ili PR-R-, HER-2-) tip karcinoma dojke. U oko 20 do 30% karcinoma dojke javlja se amplifikacija HER-2 i TOP2A gena. Amplifikacija HER-2 i TOP2A gena je povezana sa lošijim ishodom bolesti, agresivnijim tokom i dobrim odgovorom pacijentkinja na terapiju Herceptinom i antraciklinima. Koamplifikacija HER-2 i TOP2A gena je prisutna u 12 do 38% karcinoma dojke. Na osnovu rezultata nekoliko studija postavljena je hipoteza po kojoj se amplifikacija TOP2A gena javlja isključivo sa amplifikacijom HER-2 gena, mada postoje i drugačija, suprotna mišljenja. Cilj: Cilj naše studije bio je da se odrede genski statusi HER-2 i TOP2A kao i njihova korelacija sa apoptotskim i proliferativnim indeksom, nivoom ekspresije ER-R, PR-R i HER-2 receptora, TOP2A, BRCA1, Bcl-2 i p53 proteina, osnovnim kliničko-patološkim parametrima kod tumora dojke 60 pacijentkinja podeljenih u četiri molekularna tipa. Materijal i metode: Šezdeset tkivnih uzoraka karcinoma dojke ispitivano je imunohistohemijskom metodom na prisustvo i stepen ekspresije ER-R, PR-R, HER-2, p53, Bcl-2, TOP2A, BRCA1, ssDNA, Ki67, kao i FISH metodom na stepen genske modifikacije HER-2 i TOP2A. Stepen modifikacije HER-2 i TOP2A gena analiziran je primenom HER-2 FISH pharmDxTM kit i TOP2A FISH pharmDxTM kit (DakoCytomation). Rezultati: Kliničko-patološki podaci - Starost ispitanica se kretala u rasponu izmeñu 38 i 87 godina (64.79 ± 21). Veličina tumora se kretala u opsegu izmeñu 0.4 i 5.5 cm, dok je histološki gradus G1 bio zastupljen kod 6 (10%) karcinoma, histološki gradus G2 kod 25 (41.7%) i histološki gradus G3 kod 17 (28.3%) karcinoma dojke..

Correlation between protein and gene expression of HER-2 and topoisomerase 2a in infiltrative ductal breast carcinoma

Uvod: Karcinom dojke predstavlja jako heterogeno oboljenje kao i jedno od najzastupljenijih malignih bolesti u opštoj ženskoj populaciji. Na osnovu prisustva i nivoa ekspresije ER-R, PR-R i HER-2 receptora ostvarena je podela na četiri molekularna tipa karcinoma dojke: luminalni A (ER-R+ i / ili PR-R+, HER-2-), luminalni B (ER-R+ i / ili PR-R+, HER-2+), HER-2 pozitivni (ER-R- i / ili PR-R-, HER-2+) i trostruko negativni (ER-R- i / ili PR-R-, HER-2-) tip karcinoma dojke. U oko 20 do 30% karcinoma dojke javlja se amplifikacija HER-2 i TOP2A gena. Amplifikacija HER-2 i TOP2A gena je povezana sa lošijim ishodom bolesti, agresivnijim tokom i dobrim odgovorom pacijentkinja na terapiju Herceptinom i antraciklinima. Koamplifikacija HER-2 i TOP2A gena je prisutna u 12 do 38% karcinoma dojke. Na osnovu rezultata nekoliko studija postavljena je hipoteza po kojoj se amplifikacija TOP2A gena javlja isključivo sa amplifikacijom HER-2 gena, mada postoje i drugačija, suprotna mišljenja. Cilj: Cilj naše studije bio je da se odrede genski statusi HER-2 i TOP2A kao i njihova korelacija sa apoptotskim i proliferativnim indeksom, nivoom ekspresije ER-R, PR-R i HER-2 receptora, TOP2A, BRCA1, Bcl-2 i p53 proteina, osnovnim kliničko-patološkim parametrima kod tumora dojke 60 pacijentkinja podeljenih u četiri molekularna tipa. Materijal i metode: Šezdeset tkivnih uzoraka karcinoma dojke ispitivano je imunohistohemijskom metodom na prisustvo i stepen ekspresije ER-R, PR-R, HER-2, p53, Bcl-2, TOP2A, BRCA1, ssDNA, Ki67, kao i FISH metodom na stepen genske modifikacije HER-2 i TOP2A. Stepen modifikacije HER-2 i TOP2A gena analiziran je primenom HER-2 FISH pharmDxTM kit i TOP2A FISH pharmDxTM kit (DakoCytomation). Rezultati: Kliničko-patološki podaci - Starost ispitanica se kretala u rasponu izmeñu 38 i 87 godina (64.79 ± 21). Veličina tumora se kretala u opsegu izmeñu 0.4 i 5.5 cm, dok je histološki gradus G1 bio zastupljen kod 6 (10%) karcinoma, histološki gradus G2 kod 25 (41.7%) i histološki gradus G3 kod 17 (28.3%) karcinoma dojke...Introduction: Breast cancer is a heterogeneous disease and the most common malignancy in women with increasing incidence in recent years. According to immunohistochemical expression of estrogen receptor (ER-R), progesterone receptor (PR-R) and HER-2 four different molecular subtypes of breast cancer were identified: luminal A (ER-R+ and / or PR-R+, HER-2-), luminal B (ER-R+ and / or PR-R+, HER- 2+), HER-2 positive (ER-R- and / or PR-R-, HER-2+) and triple negative (ER-R- and / or PR-R-, HER-2-) breast cancer subtypes. HER-2 and TOP2A genes are amplified in 20– 30 % of breast cancers. Amplification of HER-2 and TOP2A genes are associated with a poor prognosis and predictive response to treatment by Herceptin and anthracycline. Co-amplification of HER-2 and TOP2A has been seen in approximately 12–38 % of patients with breast cancers. A few retrospective studies have reported that TOP2A gene amplification occurs almost exclusively in conjunction with HER-2 gene amplification. Aim: The aim of our study is to investigate HER-2 and TOP2A gene aberrations and their correlation with apoptotic and proliferative indexes, Bcl-2, p53, ER-R, PR-R, HER-2, TOP2A, BRCA1, clinicopathological parameters and overall survival in four molecular subtypes of breast cancer. Material and methods: Sixty paraffin-embedded breast cancer tissue samples were studied immunohistochemically for the expression of ER-R, PR-R, HER-2, p53, Bcl-2, ssDNA, Ki67, BRCA1, TOP2A and FISH for HER-2 and TOP2A gene aberrations. HER-2 and TOP2A gene copy number were analyzed by HER-2 FISH pharmDxTM kit and TOP2A FISH pharmDxTM kit (DakoCytomation), respectively. Results: Clinicopathological parameters - The ages of patients ranged from 38 to 87 years (64.79 ± 21). The tumor size ranged between 0.4 and 5.5 cm, and the histological grades were G1, G2 and G3 in 6 (10 %), 25 (41.7 %), and 17 (28.3 %) patients, respectively. In 26 patients (43.4 %) lymph node status was negative; while in 22 patients (36.6 %) was detected positive lymph node status..

On the brachistochrone of a variable mass particle in general force fields

The problem of the brachistochronic motion of a variable mass particle is considered. The particle moves through a resistant medium in the field of arbitrary active forces. Beginning from these general assumptions, and applying Pontryagin's minimum principle along with singular optimal control theory, a corresponding two-point boundary value problem is obtained and solved. The solution proposed involves an appropriate numerical procedure based upon the shooting method. In this numerical procedure, the evaluation of ranges for unknown values of costate variables is avoided by the choice of a corresponding Cartesian coordinate of the particle as an independent variable. A numerical example assuming the resistance force proportional to the square of the particle speed is presented. A review of existing results for related problems is provided, and it can be shown that these problems may be regarded as special cases of the brachistochrone problem formulated and solved in this paper under very general assumptions by means of optimal control theory

Etiopatogeneza, dijagnoza i terapija stečenog megakolona pasa

Megacolon refers to an abnormal dilatation of the colon. This condition occurs in both humans and animals. Although it seems to be more common in cats, megacolon may also occur in dogs. However, data regarding the etiopathogenesis, clinical course and outcome of canine megacolon are scarce. The aim of this study is to present the experience of our team in diagnosis and therapy of canine acquired megacolon, with particular reference to etiopathogenetic aspects. The prospective study included 28 dogs affected with megacolon, aged 5-9 years. The 26 animals underwent a surgical procedure (colonotomy followed by manual extraction of faeces), and were followed up for a period of 28 days. On the basis of anamnestic data, clinical and radiographic findings, 7 dogs (25%) were presented with idiophatic acquired megacolon, while 75% of cases had secundary acquired megacolon of different etiology (including pelvic canal stenosis, lumbar and sacral spinal injuries or back leg fractures, in 46% of cases; keeping the animals in the backyard and irresponsibility of their owners, in 11%; non-adequate nutrition, in 11%; and decreased physical activity and keeping animals in small flats, in 7%). During early postoperative period, the medical treatment and dietary regimen enabled defecation in 65% of cases. The remaining 35% of cases were treated with Cisapride in order to establish spontaneous defecation. All dogs recovered completely during the 28- days follow-up period. According to the results of interviews with dog owners, all animals were in good condition six months after the surgical procedure.Megakolon označava abnormalnu dilataciju kolona. Javlja se i kod ljudi i kod životinja. Iako je oboljenje češće kod mačaka, javlja se i kod pasa. Međutim, podaci o etiopatogenezi, kliničkom toku i ishodu bolesti megakolona kod pasa su oskudni. Stoga je cilj ove studije da se iznesu sopstvena iskustva o dijagnostici i terapiji stečenog megakolona kod pasa, sa posebnim naglaskom na etiopatogenetski aspekt oboljenja. Studija je bila prospektivnog karaktera i obuhvatila je 28 pasa sa megakolonom, starosti 5-9 godina. Dvadeset šest životinja podvrgnuto je hirurškoj intervenciji (kolonotomija sa manuelnom ekstrakcijom fecesa) i praćeno postoperativno u toku 28 dana. Na osnovu anamnestičkih podataka dobijenih od vlasnika pasa, kliničkog i radiografskog nalaza, 7 pasa (25%) imalo je idiopatski stečeni megakolon, dok je kod 21 psa (75%) bolest okarakterisana kao sekundarni stečeni megakolon uzrokovan različitim etiološkim faktorima (uključujući stenozu pelvičnog kanala, oštećenje lumbalne i sakralne kičme ili frakturu zadnjih ekstremiteta, kod 46% pasa; čuvanje životinja u dvorištu i nedovoljni nadzor od strane vlasnika, kod 11%; neadekvatna ishrana, kod 11%; smanjenja fizičke aktivnosti i čuvanje životinja u skučenom prostoru, kod 7%). U ranom postoperativnom periodu, terapija i režim ishrane doveli su do uspostavljanja defekacije kod 65% pasa. Kod preostalih 35% slučajeva spontana defekacija je uspostavljena primenom terapije Cisapridom. Sve životinje su se u toku 28-dnevnog nadzora kompletno oporavile, a odlično opšte stanje zabeleženo je, na osnovu intervjua sa vlasnicima i 6 meseci nakon hirurške intervencije

Nitric Oxide Synthase Dependency in Hydroxyurea Inhibition of Erythroid Progenitor Growth

Hydroxyurea (HU) causes nitric oxide (NO) bioactivation, acting as both a NO donor and a stimulator of NO synthase (NOS). To examine whether HU effects are NO mediated by chemical degradation or enzymatic induction, we studied human and mouse erythroid cells during proliferation, apoptosis, and differentiation. The HU and NO donor demonstrated persisted versus temporary inhibition of erythroid cell growth during differentiation, as observed by γ-and β-globin gene expression. HU decreased the percentage of erythroleukemic K562 cells in the G2/M phase that was reversed by N-nitro l-arginine methyl ester hydrochloride (L-NAME). Besides activation of endothelial NOS, HU significantly increased apoptosis of K562 cells, again demonstrating NOS dependence. Administration of HU to mice significantly inhibited colony-forming unit-erythroid (CFU-E), mediated by NOS. Moreover, burst-forming-units-erythroid (BFU-E) and CFU-E ex vivo growth was inhibited by the administration of nitrate or nitrite to mice. Chronic in vivo NOS inhibition with L-NAME protected the bone marrow cellularity despite HU treatment of mice. NO metabolites and HU reduced the frequency of NOS-positive cells from CFU-E and BFU-E colonies that was reverted by NOS inhibition. HU regulation of the G2/M phase, apoptosis, differentiation, cellularity, and NOS immunoreactive cells was NOS dependent. Inhalation of NO therapy as well as strategies to increase endogenous NO production could replace or enhance HU activity

Enzimi zaštite od oksidacionih oštećenja u toku klijanja dve linije omorike [Picea omorika (Panč.) Purkyně]

dTwo lines of Picea omorika (Pane.) Purkyne were compared with respect to germination percentage as well as specific activity and isoenzyme pattern of catalase, superoxide dismutase, and peroxidase (POD) during germination. Line A had a higher gennination percentage and higher enzyme activities in dry seeds and seedlings compared to line C. Peroxidase activity was not detected in dry seeds, but measured up to 10 U/g and 28 U/g on the 7(th) day of germination in lines C and A, respectively. The most abundant POD basic isoform in seedlings of both lines (pl 8.2) was not found previously in needles of adult Serbian spruce trees of the same lines.Poređeni su procenat klijanja, specifična aktivnost i izoenzimski profil katalaze, superoksid dismutaze i peroksidaze u dve linije omorike [Picea omorika (Panč.) Purkyně] u toku klijanja. Linija A je imala veći procenat klijanja i veće enzimske aktivnosti u suvom semenu i klijancima, u poređenju sa linijom C. Peroksidazna aktivnost nije nađena u suvim semenima, a povećavala se do 10 U/g i 28 U/g sed- mog dana klijanja u linijama C i A, respektivno. Najzastupljenija bazna izoforma peroksidaze u klijancima obe linije (pI 8.2) nije nađena ranije u četinama odraslih jedinki istih linija.

Neuronal Nitric Oxide Synthase Mediates the Effect of Ethanol on IgA

Aims: We showed previously that the acute effect of ethanol on intestinal immunoglobulin A (IgA) expression might be mediated by endogenous nitric oxide (NO). To extend these findings, this study was designed to investigate a possible role of neuronal NO synthase (nNOS) in the observed phenomenon, using 7-nitroindazole (7-NI), a selective inhibitor of its activity. Methods: Adult male Wistar rats were treated with: (a) ethanol (4 g/kg, intraperitoneally, i.p.), (b) 7-NI (25 mg/kg, i.p.) followed by ethanol (4 g/kg, i.p.) 30 min later and (c) 7-NI (25 mg/kg, i.p.) followed by saline 30 min later. Untreated rats were used as controls. The concentrations of serum and intestinal IgA were measured by enzyme-linked immunosorbent assay, while the expression of nNOS was determined using western blot and immunohistochemistry. Results: Acute ethanol treatment significantly increased the concentration of IgA in the ileal extracts, whereas it decreased its serum level. Inhibition of nNOS activity by 7-NI abolished this action of alcohol on IgA. Additionally, western blot analysis revealed that the acute alcohol administration induced an increase in the expression of intestinal nNOS. Furthermore, nNOS-immunoreactive cells, observed within the lamina propria of small intestine, were numerous in ethanol-treated rats. Conclusion: Taken together, these results extended our previous findings suggesting that nNOS mediates the acute effect of ethanol on IgA and supported an immunomodulatory role of this enzyme isoform

Ispitivanje dejstva persirćetne kiseline na lipidnu komponentu bakterijskih spora i doprinos standardizaciji testa za ocenu efikasnosti

The aim of this research was to contribute to a more detailed insight in the action of peroxygen disinfectants based on peroxoacetic acid (PAA), on lipid components of sporogenic forms of microorganisms. Sporogenic forms are recognized to be significantly more resistant in comparison to vegetative forms. Bacillus cereus ATCC 11778 and Bacillus subtilis NCTC 10480 where chosen as referent bacterial sporogenic strains used to investigate the acting mechanism of PAA. After treatment of the above mentioned bacterial strains with PAA, fatty acids have been isolated from untreated and treated spores and consequent changes in the lipid component were analyzed. The obtained methyl-esters of fatty acids (MEFA) were analyzed by gas chromatography/mass spectrometry (GC/MS), using a standard of bacterial MEFA. Results confirmed that after treatment, in Bacillus cereus certain quantitative changes occurred, which included the decrease in quantity of fatty acids with 16 and 17 carbon atoms, i.e. 16:0, 16:1, /so 17:0 and anteiso 17:0. At the same time, in B. Subtilis the significant decrease in quantity of fatty acids with 15 and 17 carbon atoms occurred, i.e. anteiso 15:0 and anteiso 17:0.Cilj ovog istraživanja je bio da se stekne potpuniji uvid u dejstvo peroksigenih dezinfektanata na bazi peroksosircetne kiseline (u daljem tekstu: PSK) na lipidnu komponentu sporogenih formi mikroorganizama, za koje je poznato da pokazuju veću otpornost od vegetativnih oblika. Bacillus cereus ATCC 11778 i Bacillus subtilis NCTC 10480 su odabrani kao referentni sojevi - bioindikatori, na kojima je ispitivano delovanje PSK (nakon formiranja spora). Nakon tretmana spora pomenutih bakterijskih sojeva sa PSK, izolovane su masne kiseline iz netretiranih i tretiranih spora i analizirane su promene u lipidnoj komponenti. Dobijeni metil-estri masnih kiselina (MEMK) analizirani su pomoću gasne hromatografije i masene spektrometrije (GC/MS) uz poređenje sa standardom bakterijskih MEMK. Dobijeni rezultati su ukazali da je kod B. cereus ATCC 11778 nakon tretmana došlo do kvantitativne promene odnosno smanjenja količine masnih kiselina sa 16 i 17 Catoma, odnosno 16:0,16:1, ante/so 17:0,/so 17:0. Kod B. subtilis NCTC 10480 došlo je do značajnog smanjenja količine masnih kiselina sa 15 i 17 C atoma, odnosno anteiso 15:0 i anteiso 17:0

Brachistochronic motion of a variable mass system

A mechanical system consisting of rigid bodies and material particles, of which some particles are with variable masses, is considered. Laws of variation of the masses of the points and relative velocity of particles separating from the points are well-known. The system is moving in an arbitrary field of known potential and nonpotential forces. Applying Pontryagin’s Maximum Principle and singular optimal control theory, brachistochronic motion is determined. A two-point boundary value problem, due to nonlinearity of equations in a general case, is needed to be solved using some of the numerical procedures. Here the Shooting method is used, where the missing boundary conditions are chosen so as to be the physical variables (velocity and mass). The field where they are found can be approximately estimated, which is not the case with the conjugate vector coordinates being of purely mathematical nature. The paper also presents the manner of brachistrochronic motion realization without the action of active control forces. It is realized by subsequent imposition to the system a corresponding number of independent ideal holonomic mechanical constraints. The constraints must be in accordance with the previously determined brachistochronic motion of the system. The method is illustrated by an example of determining the brachistochronic motion of the system with three degrees of freedom and method of its realization. The system consists of one rigid body to which two points of variable masses are attached, where the system is moving in a vertical plane. Brachistochronic motion is realized by the help of two ideal holonomic constraints

Lysiphlebus orientalis (Hymenoptera, Braconidae), a new invasive aphid parasitoid in Europe - evidence from molecular markers

We report the occurrence of Lysiphlebus orientalis in Serbia, an aphid parasitoid from the Far East that is new to Europe and has the potential to become invasive. Our finding based on morphological characters is confirmed by analyses of mitochondrial cytochrome oxidase subunit I sequences. An increase in number and an expansion of the host range were observed during field studies over the past two years, and it is determined that the current host range encompasses nine aphid hosts on 12 different host plants, forming 13 tri-trophic associations. A host range determined for European populations of L. orientalis appears wider compared with that in its Far Eastern native habitats where Aphis glycines Mats. is the sole known host. Moreover, it overlaps considerably with the host ranges of European parasitoids that play an important role in the natural control of pest aphid