Biological markers in breast cancer prognosis and treatment.

In this review we will provide a synopsis of the biological markers used in the care of breast cancer patients with emphasis on clinical application. The advent of molecular technology has incorporated new biomarkers along with the older immunohistochemical and serum ones. Serum tumor markers are proteins shed from breast cancer cells. Their levels have long been used as a measure of tumor burden and disease progression or recurrence. However, limitations exist that should be known to those involved in breast cancer management. Historically, immunohistochemical markers have been used to guide treatment decisions. These markers reveal characteristics of the cancer cells and have been used both as prognostic and predictive factors. Molecular markers give information on the expression of certain genes in tumor tissues related to proliferation, invasion, and metastasis and researchers try to correlate them with the use of mathematical modeling with clinical outcomes, hence those markers exhibit prognostic and predictive significance. All these tools can guide personalized treatment by estimating patient prognosis and risk of relapse and tailor accordingly therapeutic approaches.

Targeted treatment for metastatic renal cell carcinoma and immune regulation.

New targeted agents have become the mainstream of treatment in metastatic renal cell carcinoma (mRCC) and substituted the previous cytokine-based therapies. Vascular endothelial growth factor (VEGF) pathway is the principle target for drugs like sunitinib, sorafenib and bevacizumab. As VEGF is regulating dendritic cell (DC) function, inhibition of VEGF results in activation of DCs and a shift towards cellular (type 1) immunity, which is believed to favor cancer rejection. Recent studies have established the immune-stimulating effects of sunitinib that may as well be a marker for effectiveness. On the other hand, sorafenib not only inhibits VEGF receptor (VEGFR) but is also a B-Raf inhibitor (a component of the ras - MAPK pathway) and this leads to downregulation of immune responses. Sorafenib has not yet shown benefit in first-line treatment of mRCC when compared to interferon (IFN)-alpha and sorafenib-mediated immunosuppression may partially account for that. Mammalian target of rapamycin (mTOR), the target of temsirolimus, is an element of the DC activation pathway. There are no data for in vivo effects of temsirolimus in the immune system. The addition of IFN-alpha to temsirolimus resulted in inferior outcomes than temsirolimus alone. IFN-alpha has however still a place in mRCC treatment, as bevacizumab has been approved in combination with IFN-alpha. New clinical trials address the effects of the combination of cytokines with targeted agents. The immune-modulating effects of targeted treatments may be important in pharmacodynamic outcomes, effectiveness or the development of adverse events.

Effect of HbS in the determination of HbA2 with the Menarini HA-8160 analyzer and comparison with other instruments.

It is known that the presence of hemoglobin S (HbS) affects the determination of hemoglobin A(2) (HbA(2)) levels in clinical samples. We quantitated this effect using the Menarini HA-8160 analyzer and compared with other instruments (HELENA beta-thal quik column, TOSOH HLC-723G7 and BIORAD Variant II) using the HELENA SAS-MX alkaline gel electrophoresis kit as the reference method. The %HbA(2) values from the HA-8160 analyzer and the alkaline gel electrophoresis show a good linear correlation in the absence of HbS. A strong positive bias in the %HbA(2) values from the HA-8160 is apparent in the presence of HbS in the samples, when compared with the alkaline electrophoresis. The analytical imprecision and bias of the three HPLC instruments are comparable both in the presence and absence of HbS. The manual column method shows a lower bias in the absence of HbS but is more affected when HbS is present in the samples.

Cell signaling in cancer.

The pathways that control cell differentiation and growth are almost always compromised in cancer. Although in the last years there have been major advances in understanding these changes and how they contribute to tumor initiation and growth, the task is far from complete. In this review we discuss some of the factors that are found in major key nodes of the signaling pathways. Included among them are the nuclear factor kappaB (NF-kappaB) that has a quite central role in inflammation, and the mammalian target of rapamycin (mTOR) kinase. Also an eminent role is played by the EGF (epidermal growth factor) and the Snail/Slug family of repressors. Since the expansion of tumor cells depends heavily on nutrient and oxygen supply, it requires the growth of new blood vasculature which is directed by the hypoxia inducible factor (HIF).

The unexpected formation of biologically active Cu(II) Schiff mono-base complexes with 2-thiophene-carboxaldehyde and dipropylenetriamine: crystal and molecular structure of CudptaSCl2.

Two novel mononuclear Cu(II) coordination compounds of the type [Cu(dptaS)Cl(2)] and [Cu(dptaS)Br(2)] (dptaS=1,3-propanediamine, N(1)-[3-aminopropyl]-N(3)-[2-thienylmethylidene] or Schiff mono-base of dipropylenetriamine with 2-thiophene-carboxaldehyde) were prepared by the hydrolysis of the di-bases, [Cu(dptaSS)Cl(2)] and [Cu(dptaSS)Br(2)] (dptaSS=1,3-propanediamine, N(1)-[2-thienylmethylidene]-N(3)-[[2-thienylmethylidene]aminopropyl] or Schiff di-base of dipropylenetriamine with 2-thiophenecarboxaldehyde) to mono-bases with the release of one aldehyde molecule and freeing of the -NH(2) group of the coordinated dpta ligand. The X-ray determined structure of the compound [Cu(dptaS)Cl(2)] was confirmed by spectroscopic methods, magnetic and molar conductivity measurements. The Cu(II) atom is a five-coordinated CuN(3)Cl(2) chromophore with three nitrogen atoms coming up from the (dptaS) ligand and two chlorine atoms completing the square pyramidal geometry. Antiproliferative activity of both novel compounds was examined against a panel of different normal and cancer cell lines (MRC5, HeLa, MCF7, HT-29 and T47D) and showed that the Cu(II) Schiff mono-bases exhibit increased activity as compared to the starting materials. In vitro studies of plasmid DNA (pDNA) and double stranded DNA (dsDNA) interaction with the compounds under study support this difference. Some of the important factors contributing to the antiproliferative activity of the compounds under study, such as ionic character and dipole moment were also discussed in terms of the density functional theory calculated electronic structures of the ligands and their Cu(II) compounds.

Copper(II) Schiff base coordination compounds of dien with heterocyclic aldehydes and 2-amino-5-methyl-thiazole: synthesis, characterization, antiproliferative and antibacterial studies. Crystal structure of CudienOOCl2.

A new series of coordination compounds of the starting materials [Cu(dienX(2)Y(2))] and their adducts [Cu(dienXXY(2))(2a-5mt)] (where dien=diethylenetriamine, dienXX=Schiff bases of diethylenetriamine with 2-furaldehyde or 2-thiophene-carboxaldehyde, X=O, S, Y=Cl, Br, NO(3) and 2a-5mt=2-amino-5-methylthiazole) were synthesized by stepwise reactions and their structures were established by C, H, N, Cu analysis, spectroscopic, magnetic and molar conductivity measurements. The isolated compounds are monomers, paramagnetic and electrolytic compounds of the type 1:1. In all cases, the pentadentate Schiff base (dienXX) is bonded in a tridentate fashion through the 3 N atoms. In the CudienXXY(2) compounds the coordination sphere is completed by two Cl or Br or NO(3) groups in a square pyramidal arrangement. The proposed structure for this type of compound was further supported by X-ray diffraction analysis of the compound [Cu(dienOO)Cl(2)]. Its basal plane consists of three Cu-N contacts [2.017(2), 2.025(2) and 2.012(2) A] from dienOO, and the Cl(1) atom, while the Cl(2) atom possesses the apical position, the relevant distances being 2.2732(7) A for Cu-Cl(1) and 2.6051(7) A Cu-Cl(2). In the CudienX(2)Y(2).2a-5mt adducts the coordination sphere of copper is further completed by the nitrogen ring atom of the 2a-5mt, forming an octahedral configuration. The study of the biological activity of the compounds synthesized against a panel of different normal and cancer cell lines (MRC5, HeLa, MCF7, HT-29, OAW42, T47D) and bacteria (E. coli, B. cereus, B. subtilis) showed that the adducts of the type [Cu(dienXXY(2))(2a-5mt)] exhibit increased activity both in cancer cells and in bacteria, compared to the starting material of type [Cu(dienXXY(2))].

Exercise-induced changes in c-Fos protein levels in skeletal muscle of trained and untrained rats.

We investigated the effect of an acute bout of endurance exercise on c-Fos protein levels in the extensor digitorum longus muscle of trained and untrained rats. Fifty rats were equally divided into a trained and an untrained group. Rats of the trained group ran on a treadmill 45 min/day for 5 days. On the sixth day, 5 rats were killed without exercise, while the remaining 20 ran as above and were killed 0, 3, 6, and 12 h post-exercise (5 rats at each time point). In the untrained group, 5 rats were killed without exercise, while the remaining 20 ran as above only once and were killed at the same time points as the trained group. Western blotting demonstrated no significant changes in c-Fos protein levels in the untrained group. On the contrary, in the trained group, there was a significant increase at 6 and 12 h compared to 3 h post-exercise. The levels of the protein in the trained rats were above the corresponding levels in the untrained ones at all time points, although these differences reached statistical significance only immediately, 6 h and 12 h post-exercise. These results show that trained skeletal muscle exhibits increased levels of c-Fos, probably as a cumulative result of changes occurring during recovery from each exercise bout, and greater c-Fos response after acute endurance exercise compared to untrained skeletal muscle.

Myeloma metastasis to the testis: findings on gray-scale and color Doppler ultrasonography with literature review.

Testicular metastasis of multiple myeloma is very rare and only 49 cases have been reported in the world literature. We report on a case of a 59-year-old man with a 9- month history of multiple myeloma who presented with painless swelling of the left testis. An ultrasound (US) examination was performed and the diagnosis was documented histologically. The US findings and the differential diagnosis are discussed in the light of color Doppler ultrasonography (CDUS).

Antiproliferative activity of mixed-ligand dien-Cu(II) complexes with thiazole, thiazoline and imidazole derivatives.

The reaction of [Cu(dien)NO(3)]NO(3) with 2-amino-5-methylthiazole (2A5MT), 2-amino-2-thiazoline (2A-2Tzn), imidazole (im), N,N'-thiocarbonyldiimidazole (Tcdim), 2-aminothiazole (2AT) and 2-ethylimidazole (2Etim), gave a new series of mixed-ligand compounds of the general formula [Cu(dien)(B)NO(3))]NO(3); (dien, diethylenetriamine; B, 2A5MT, 2A-2Tzn, im, Tcdim, 2AT and 2Etim). The complexes have been characterised by elemental analysis, molar conductivity and magnetic measurements, as well as by electronic and IR spectral studies. According to the above measurements the possible structure of the compounds is the square pyramidal in the solid state and the square planar in aqueous solution. We tested all complexes for antiproliferative (cytostatic and cytotoxic) activity against a panel of cell lines (HeLa, L929, HT-29 and T47D). All [(dien)Cu(B)NO(3))](NO(3)) complexes had an activity against colon cancer cells (HT-29), inducing G2/M cell cycle arrest, an effect that for most of the complexes could be attributed to p34cdc2 inhibition by tyrosine-phosphorylation and/or to induction of (cyclin-dependent kinase inhibitor) p21(WAF1). Other cell lines were resistant to the majority of the complexes, except [Cu(dien)(2A5MT)NO(3))](NO(3)), that had showed the highest anti-proliferative activity against HT-29 cells also. The predilection for colon cancer cells and the relatively low toxicity against normal (L929) cells justify further investigation of this group of compounds.

In vitro cytotoxicity of six dentin bonding agents.

The cytotoxicity of six dentin bonding agents (Syntac, Solobond, Bond 1, Scotchbond 1, Heliobond and F-2000) was tested against an established cell line, L929. Under aseptic conditions 3, 5 and 10 microL dentin bonding agents were placed in the centre of Petri dishes. Each dish was covered with a 5-mL suspension of fibroblasts at a concentration of 40 000 cells mL(-1). The cultures were incubated at 37 degrees C and cytotoxicity was assessed by a quantitative technique at 24 and 72 h. All the dentin bonding agents were found to be cytotoxic. Scotchbond 1 and F-2000 showed the highest cytotoxicity followed by Solobond and Bond 1. Heliobond and Syntac were the least toxic materials.

Antitumor activity of L-asparaginase from Thermus thermophilus.

L-asparaginase (EC 3.5.1.1) was purified to homogeneity from Thermus thermophilus. The apparent molecular mass of L-asparaginase was found to be 33 kDa by SDS-PAGE, whereas by Sephacryl S-300 superfine column it was found to be 200 kDa, indicating that the enzyme in the native stage acts as hexamer. It is a thermostable enzyme and keeps all of its activity at 80 degrees C for 10 min. The antiproliferative activity of the purified L-asparaginase from T. thermiphilus was tested against the following human cell lines: K-562 (chronic myelogenous leukemia), Raji (Burkitt's lymphoma), SK-N-MC (primitive neuroectodermal tumor), HeLa (cervical cancer), BT20 and MCF7 (breast cancers), HT-29 (human colon cancer), and OAW-42 (ovarian cancer). The antiproliferative activity of T. thermophilus enzyme was compared with Erwinase, the commercially available L-asparaginase from Erwinia corotovora. The potency difference between the two L-asparaginases was greater in HeLa and SK-N-MC than in other cell lines. The fact that L-asparaginase from T. thermophilus does not hydrolyse L-glutamine makes it advantageous for future clinical trials.

Protein tyrosine kinase inhibitor, genistein, enhances apoptosis and cell cycle arrest in K562 cells treated with gamma-irradiation.

Genistein, is a natural isoflavone compound with a potent activity against protein tyrosine kinases. The leukemic cell line, K562, is a bcr/abl (Philadelphia chromosome) positive cell line that is resistant to DNA-damaging agents, including gamma-irradiation. Treatment with genistein increased apoptosis and promoted G2-phase arrest in the non-apoptotic population of the gamma-irradiated K562 cells. Irradiated cells that survived 72 h after the irradiation had a normal distribution in cell cycle, whilst genistein treatment kept cells arrested in the G2-phase, decreased the S-phase fraction and suppressed DNA-synthesis. Taken together, our results show that genistein augments apoptotic cell death after gamma-irradiation in K562 cells and this result cannot be attributed to abrogation of the G2/M checkpoint.

CDK-inhibitor olomoucine inhibits cell death after exposure of cell lines to cytosine-arabinoside.

Signal transduction for apoptosis or programmed cell death, after DNA damage in mammalian cells, is believed to involve activation of cyclin-dependent kinases (CDKs), especially CDK-1 (cdc2) and CDK-2. We used CDK-inhibitor olomoucine, a purine analogue to evaluate the role CDK inhibition on cytosine-arabinoside (Ara-C)-induced cell death. The two drugs showed an antagonistic effect, suggesting that apoptosis after exposure to Ara-C is inhibited by olomoucine. DNA-electrophoresis showed a clear inhibition of the apoptotic pattern when olomoucine was added to Ara-C. We conclude that CDK-inhibitor olomoucine inhibits cell death induced by Ara-C.

Cytotoxic effects of different concentrations of neutral and alkaline EDTA solutions used as root canal irrigants.

The cytotoxic effects of neutral and alkaline EDTA solutions were evaluated and compared with those of sodium hypochlorite solution using an established cell line: L929. Cytotoxicity was assessed by a quantitative technique at five observation periods (1, 3, 6, 12, and 24 h). All tested agents showed moderate to severe cytotoxicity in the present experimental model in a concentration-dependent manner.

In vitro release of hydroxyl ions from calcium hydroxide gutta-percha points.

In endodontic practice, calcium hydroxide is widely used for a number of reasons associated with its high pH. The purpose of the present study was to determine in vitro the alkalizing potential of newly introduced calcium hydroxide gutta-percha points that are proposed for temporary filling of root canals. The materials tested were: calcium hydroxide gutta-percha points; chemical pure calcium hydroxide powder mixed with distilled water; and Reogan rapid, a nonsetting calcium hydroxide preparation. The materials were placed into dialysis tubing and transferred into plastic vials containing bidistilled water. Measurements were taken by a digital pH meter after 10, 20, and 30 s; 1, 15, and 30 min; and 1, 2, 3, 24, 48, 72, 96, and 120 h. The calcium hydroxide containing gutta-percha points showed a significantly lower alkalizing potential than Reogan rapid and calcium hydroxide mixed with distilled water (p < 0.05).

Anti-inflammatory drugs interacting with Zn(II), Cd(II) and Pt(II) metal ions.

Complexes of Zn(II), Cd(II) and Pt(II) metal ions with the anti-inflammatory drugs, 1-methyl-5-(p-toluoyl)-1H-pyrrole-2-acetic acid (Tolmetin), alpha-methyl-4-(2-methylpropyl)benzeneacetic acid (Ibuprofen), 6-methoxy-alpha-methylnaphthalene-2-acetic acid (Naproxen) and 1-(4-chlorobenzoyl)-5-methoxy-2-methyl-1H-indole-3-acetic acid (indomethacin) have been synthesized and characterized. In the structurally characterized Cd(naproxen)2 complex the anti-inflammatory drugs acts as bidentate chelate ligand coordinatively bound to metal ions through the deprotonated carboxylate group. Crystal data for 1: [C32H26O8Cd], orthorhombic, space group P22(1)2(1), a = 5.693(2) (A), b = 8.760(3) (A), c = 30.74(1) (A), V = 1533(1) A3, Z = 2. Antibacterial and growth inhibitory activity is higher than that of the parent ligands or the platinum(II) diamine compounds.

Cytotoxicity of three resin-based root canal sealers: an in vitro evaluation.

The cytotoxicity of three resin-based root canal sealers (AH26, AH-Plus, Topseal) was evaluated in vitro. The experiments included two cell lines, L929 mouse skin fibroblasts and RPC-C2A rat pulp cells. The cytotoxicity was assessed by sulforodamine B (SRB) colorimetric assay and hemocytometer viable cell counting after 24- and 48-h exposure. AH26 had a severe cytotoxic effect whilst Topseal and AH-Plus showed a markedly lower toxic influence on the cells during the experimental period.

In vitro combined effect of a new series of copper (II) complexes with cisplatin or epirubicin on human breast and cervical cancer cell lines.

The in vitro cytotoxic effects of some recently synthesized copper (II) complexes were investigated in combination with two clinically important anticancer agents, cisplatin (CDDP) and epirubicin (EPR), on three human breast cancer cell lines (BT20, MCF7 and T47D), a human cervical carcinoma cell line (HeLa) and two non-tumour cell lines (BHK-21 and L929). The in vitro antiproliferative effects were measured by means of XTT assay and drug potency was expressed in terms of IC50 values. Synergistic effects were observed for EPR in combination with a Cu(II) malonate derivative in BT20, MCF7 and HeLa cells. Antagonistic effects were apparent when CDDP combined with a pivalate derivative in HeLa cells. For other drug combinations, synergism, adjuvance or antagonism was demonstrable depending on the cell line applied or the varied administration schedules. The results suggest that these novel complexes may enhance the cytotoxic activity of CDDP or EPR against human tumour cell lines when administered in combination with them.

In vitro evaluation of the cytotoxicity of a bleaching agent.

Cell cultures of L929 and BHK21/C13 cells were used to evaluate the toxicity of a newly introduced bleaching agent (Colgate Platinum) compared to hydrogen peroxide, an established bleaching agent. The cell reaction was determined by a quantitative technique at 24 h and 72 h. Both bleaching materials had a dose-dependent effect on cell viability. Concentrations of hydrogen peroxide causing a 50% decrease in cell number (50% inhibition dose-ID50) were calculated as 0.00034% after 24 h and 0.00001% after 72 h in L929 cells. The ID50 of hydrogen peroxide was found to be 0.00016% after 24 h and 0.00007% after 72 h in BHK21/C13 cells. The ID50 of Colgate Platinum was 0.00074% after 24 h and 0.00045% after 72 h in L929 cells and 0.00055% after 24 h and 0.00024% after 72 h in BHK21/C13 cells. The results showed that, in vitro, both bleaching agents were cytotoxic to fibroblasts and the new bleaching agent was less toxic than hydrogen peroxide.

Antiproliferative activity of synthetic tetrapeptides, analogs of AS-I phytotoxin, towards cancer cell lines.

The in vitro chemosensitivity of three cancer cell lines [HT29 (colon), HeLa (cervical) and T47D (breast)] to eight synthetic tetrapeptides, analogs of AS-I toxin, with phytotoxic effect on a series of plants was studied. Mouse fibroblast L929 cell line was also tested for chemosensitivity to these peptides. All cell lines were especially sensitive to Cys-Val-Gly-Glu tetrapeptide with IC50 values of 0.18, 0.3 and 0.63 mM for HT29, HeLa and T47D cells, respectively, whereas the IC50 value for the L929 cells was higher than 1 mM. Antiproliferative activity was also observed with peptides Tyr-Val-Gly-Glu and His-Val-Gly-Glu with IC50 values higher than those obtained for Cys-Val-Gly-Glu. For the rest of the peptides tested the IC50 values were found close to or higher than 3 mM.