Targeting the Oxytocin Receptor for Breast Cancer Management: A Niche for Peptide Tracers.

Breast cancer is a leading cause of death in women, and its management highly depends on early disease diagnosis and monitoring. This remains challenging due to breast cancer's heterogeneity and a scarcity of specific biomarkers that could predict responses to therapy and enable personalized treatment. This Perspective describes the diagnostic landscape for breast cancer management, molecular strategies targeting receptors overexpressed in tumors, the theranostic potential of the oxytocin receptor (OTR) as an emerging breast cancer target, and the development of OTR-specific optical and nuclear tracers to study, visualize, and treat tumors. A special focus is on the chemistry and pharmacology underpinning OTR tracer development, preclinical in vitro and in vivo studies, challenges, and future directions. The use of peptide-based tracers targeting upregulated receptors in cancer is a highly promising strategy complementing current diagnostics and therapies and providing new opportunities to improve cancer management and patient survival.

The impact of flushing with pre-filled saline syringes on the incidence of peripheral venous catheter failure: A quasi-experimental study.

BACKGROUND: Short peripheral venous catheters are one of the most frequently used devices in hospitals. Peripheral venous catheter failure, defined as the unscheduled dysfunction of peripheral venous catheter, is common and frequently entails a new invasive procedure. Flushing the catheter maintains patency and could prolong peripheral venous catheter dwell time. The introduction of pre-filled saline flushing syringes as compared to manually filled saline flushing syringes could facilitate the frequency of catheter flushing, and subsequently it could reduce peripheral venous catheter failure rate. OBJECTIVE: To demonstrate differences in overall peripheral venous catheter failure rates before and after the introduction of pre-filled saline flushing syringes and to assess the risk factors for peripheral venous catheter failure. METHODS: Quasi-experimental design, before-and-after intervention study. Intervention: introduction of pre-filled saline syringes for flushing. Multicenter study conducted in medical and surgical wards of three European hospitals during a 9-month period (4 months pre-intervention, 5 months intervention). A multivariate Cox proportional model was used to identify factors associated with the occurrence of peripheral venous catheter failure. RESULTS: Data from 3853 peripheral venous catheters in 1915 patients were analyzed. Compared to pre-intervention period, a significant decrease in peripheral venous catheter failure rate was observed in the intervention period (57% vs 43.4%, p < 0.001). Independent factors associated with peripheral venous catheter failure were as follows: Charlson score ⩾4 (hazard ratio: 1.648; 95% confidence interval: 1.069-2.527), days of hospital stay ⩾10 (hazard ratio: 1.468; 95% confidence interval: 1.172-1.837), and catheter "D" (hazard ratio: 1.758; 95% confidence interval: 1.058-2.919). CONCLUSION: The use of pre-filled saline syringes significantly reduced peripheral venous catheter failure and increased catheter dwell time. Thus, it is important to reinforce the use of the pre-filled syringes for flushing to reduce the incidence of peripheral venous catheters' failure.

Differential expression of two activating transcription factor 5 isoforms in papillary thyroid carcinoma.

BACKGROUND: Activating transcription factor 5 (ATF5) is a member of the activating transcription/cAMP response element-binding protein family of basic leucine zipper proteins that plays an important role in cell survival, differentiation, proliferation, and apoptosis. The ATF5 gene generates two transcripts: ATF5 isoform 1 and ATF5 isoform 2. A number of studies indicate that ATF5 could be an attractive target for therapeutic intervention in several tumor types; however, so far, the role of ATF5 has not been investigated in papillary thyroid carcinoma (PTC). METHODS: Quantitative real-time reverse transcription polymerase chain reaction and immuno-histochemical staining were used to study ATF5 mRNA and protein expression in PTC. RESULTS: We report here that ATF5 is expressed more in PTC tissue than in normal thyroid tissue. Furthermore, this is the first study that describes the presence of both ATF5 isoforms in PTC. CONCLUSION: These findings could provide potential applications in PTC cancer treatment.

The role of microRNAs in thyroid carcinomas.

Thyroid cancers (TCs) are the most common malignancies of endocrine organs. They originate from cells of different origin within the thyroid gland, which is located at the base of the neck. Several forms of TCs have been classified and great variability is observed in molecular, cellular and clinical features. The most common forms have favorable prognosis but a number of very aggressive TCs, which are characterized by a less differentiated cellular phenotype, have no effective treatment at the moment. While TC causes are not completely understood, many genetic factors involved in their onset have been discovered. In particular, activating mutations of BRAF, RET or RAS genes are known to be specifically associated with TC initiation, progression and outcome. The involvement of microRNAs in thyroid neoplasms has recently changed the paradigm for biomarker discovery in TC, suggesting that these small non-coding RNAs could be used to develop, refine or strengthen strategies for diagnosis and management of TCs. In this review, the importance of microRNA profiling in TC is explored suggesting that these molecules can be included in procedures that can perform better than any known clinical index in the identification of adverse outcomes.

Diagnostic consideration for sinonasal Wegener's granulomatosis clinically mistaken for carcinoma.

We report a case of Wegener's granulomatosis clinically mistaken for carcinoma in a 21-year-old girl presenting with an ulcerated mass of the nasopharynx associated with enlarged laterocervical nodes. The lesion was clinically suspected as malignant on the basis of clinical and radiological findings (namely, computed tomography scan and positron emission tomography). However, multiple biopsies were not conclusive for malignancy showing histological change suggestive of Wegener's granulomatosis. A serum determination of cANCA supported the diagnosis of Wegener's granulomatosis. Clinical findings and image studies suggested an erroneous diagnosis of malignancy whereas a definitive diagnosis of Wegener's granulomatosis was achieved only after repeated biopsies thus leading to a correct therapeutic approach. The Wegener granulomatosis must be added to the list of the differential diagnoses of the masses of the nasopharynx associated with or without enlarged laterocervical nodes.

Renal artery thrombosis following an inappropriate implantable cardioverter defibrillator shock.

In spite of technological advances, the incidence of inappropriate therapies continues being high. Usually, the most direct consequence of inappropriate shocks is impairment on quality of life parameters, but in some cases the consequences may be unpredictable. We report on a case of renal artery thrombosis, following an inappropriate implantable cardioverter defibrillator shock.

Temporal retinal sensitivity in mesopic adaptation.

PURPOSE: Night driving is a complex visual task with important ramifications for driver and pedestrian safety. It is usually performed under mesopic or scotopic conditions and frequently, in the presence of transient glare sources that can adapt parts of the central retina. The objective of this work was to analyze the time response of adaptation for the central 15° of the retina when part of it is exposed to transient or steady mesopic adapting fields. METHODS: Absolute visual thresholds and luminance thresholds when viewing steady and transient adaptation fields were measured for three observers, at temporal retinal eccentricities of 0°-14.5° in steps of 2.9° (subsequently described as 0°, 3°, 6°, 9°, 12° and 15°) using a two-channel Maxwellian view optical system. The adaptation field and stimulus subtended 1.05° and 0.45° respectively. The transient adaptation field was presented with a stimulus onset asymmetry (SOA) of 300 ms. Time course adaptation curves were also measured at 0°, 6° and 9°. RESULTS: The absolute dark adaptation threshold (threshold measured at dark adaptation conditions or L(a)(t) decreases in peripheral retina due to an increasing rod contribution. Luminance thresholds vs eccentricity curves for transient L(SOA300)(t) and steady L(LA)(t) mesopic adaptation fields intersect across the first 15° of the peripheral retina. CONCLUSIONS: While the fovea shows higher sensitivity than the areas of peripheral retina investigated in this study, the speed of adaptation, measured from the visibility loss, is greater for retinal regions between 6° and 9° than for the fovea or retinal eccentricities beyond 9°.

MARCH-I expression in cord blood CD34+KDR+ cells.

Hematopoietic stem cells transplantation has been successfully used in the treatment of patients with hematological malignances. A better knowledge of the mechanisms beyond their ability to completely repopulate the entire hematopoietic system would help in the treatment of hematological diseases. For this reason we focused our studies on a cell population that has been demonstrated to have some peculiar characteristics among the stem cells: CD34+KDR+ cells. These cells, an extremely rare population among the CD34 (0.1%-0.5%) cells, have been demonstrated from different groups to have the potential to give rise to the hematopoietic and endothelial lineage. By a subtraction library approach we found different sequences more expressed in CD34+KDR+ than their CD34+KDR- counterpart. In particular, we found an open reading frame correspondent to a newly characterized E3 ligase, MARCH-I. This gene is part of a recently described family involved in immune response modulation through the proteosomal mediated degradation. MARCH-I expression in stem cells could be important for their intrinsic immune properties.

Ocular tolerance of a new multipurpose solution specifically formulated for daily wear of silicone hydrogel contact lenses.

PURPOSE: To evaluate the safety, physiological performance, and effect on ocular tissues of a new multipurpose disinfecting solution (MPDS) specifically formulated for use with silicone hydrogel (SH) contact lenses (CL). METHODS: Two MPDS [solution A (Solo-care Aqua(®), Ciba-Vision) and solution B (Hidro Health(®), Disop)] were randomly assigned and prescribed in a clinical trial in a crossover clinical trial (Registered #293/07/EC; Spanish National Health Department). Only lotrafilcon B CL daily wearers were included. After each solution was used, a masked investigator assessed the slit-lamp findings (SL9, Topcon Inc.), lens deposits, and wettability according to the Guidance for clinical investigation (ISO 11980). CL daily wearing time (hours per day and days per week) were recorded. RESULTS: Fifty-four daily wearers were included. Average CL wear time was 8.3 ± 2.3 (CI95% 7.9-8.7) hours per day (8.5 ± 2.7 at baseline visit, 8.2 ± 1.9 with solution A, and 8.4 ± 2.2 with solution B). The average number of days per week of CL wear was 5.9 ± 1.3 (CI95% 5.6-6.1) (5.4 ± 1.6 at baseline visit, 6.1 ± 0.9 with solution A, and 6.0 ± 1.1 with solution B). Non-adverse slit-lamp findings were recorded (higher than 2 points). No statistical difference (P>0.05 Friedman test) in CL wearing time, number of days of CL wears and slit-lamp findings were found between the two solutions. CONCLUSIONS: This clinical trial shows that the new formulation of Hidro Health(®) MPDS is safe when used for the care of daily-wear lotrafilcon B CL. This new solution has no clinical significance on ocular tissues, according to the Guidance for clinical investigation (ISO 11980).

Circulating fibrocytes as a marker of liver fibrosis in chronic hepatitis C.

Surrogate markers of liver fibrosis are needed as an alternative to liver biopsy, which is invasive and life-threatening. Peripheral blood fibrocytes (PBF) are considered to be involved in systemic fibrogenic processes. We measured the level of PBF in patients with chronic hepatitis C by enrolling 70 patients affected with chronic hepatitis C, 20 patients with HCV-positive decompensated cirrhosis and 30 healthy volunteers. All patients underwent liver biopsy and Fibroscan for fibrosis assessment. Patients with chronic hepatitis C had significantly higher levels of PBF in comparison with healthy individuals and decompensated cirrhotics. Patients in the F0-F1 stage had a percentage of PBF of 23.3+/-4%, significantly lower (p<0.001) than in F2 and F3 stages. Patients in the F4 stage had a PBF rate of 50.6+/-2% (p<0.001 versus the F0, F1 and F2 stages). The percentage of PBF correlated positively with the Metavir score and the liver stiffness as measured by Fibroscan. PBF are increased in patients with HCV infection and correlate with the histological stage of liver fibrosis.

Differences in the daily symptoms associated with the silicone hydrogel contact lens wear.

OBJECTIVE: To determine the differences in symptoms associated with daily wear of silicone hydrogel (lotrafilcon B) contact lenses (CLs) after CL insertion and at the end of wear and their relationship with environmental factors and multipurpose solutions (Solo-care Aqua, CIBA Vision, Duluth, GA and Hidro Health, Disop, Spain). METHODS: Fifty-four patients were fitted with lotrafilcon B CLs. Two multipurpose solutions were randomly assigned and prescribed in a double-masked crossover study with three visits. After each solution was used, two questionnaires were conducted, including a survey produced by us and Contact Lens Dry Eye Questionnaire. Our questionnaire included 10 items addressing discomfort, blurry vision, lens-handling problems, dryness, redness, tearing, burning, itching, discharge, and dissatisfaction. Patients were assigned scores from 0 (without symptoms) to 10 (symptoms unbearable) at two different times (after CL insertion and at the end of wear) and in two environments (outdoors and adverse environments). RESULTS: The average duration of CL wear was 8.32 +/- 2.27 hr/day and 5.85 +/- 1.30 days/week. All symptoms became worse (P < 0.05 Wilcoxon test, except lens handling) at the end of CL wear in all visits. The results were independent of the multipurpose solution used (P > 0.05 Friedman test), except for tearing, which showed statistical differences between visits (P = 0.03 Friedman test). Contact Lens Dry Eye Questionnaire showed increased dry eye symptoms at the end of wear (P < 0.05 Friedman test). CONCLUSIONS: Daily wear symptoms associated with lotrafilcon B CL increase with the time of wear. This increase in symptoms is nonrelated with the multipurpose solutions compared in this study.

16,16-Dimethyl prostaglandin E2 efficacy on prevention and protection from bleomycin-induced lung injury and fibrosis.

In this study, we evaluated the protective effect and therapeutic potential of the prostaglandin E(2) (PGE(2)) synthetic analog 16,16-dimethyl-PGE(2) (dmPGE(2)) in the animal model of pulmonary fibrosis induced by bleomycin. Mice subjected to intratracheal administration of bleomycin (1 mg/kg) received a dmPGE(2) dose of 30 microg/kg/day by continuous subcutaneous infusion. Bronchoalveolar lavage (BAL); immunohistochemical analysis for IL-1, TNF-alpha, and nitrotyrosine; measurement of fluid content in lung; myeloperoxidase activity assay; and lung histology were performed 1 week later. Lung histology and Sircol assay for collagen deposition were performed 3 weeks after treatments. Changes of body weight and survival rate were also evaluated at 1 and 3 weeks. Compared with bleomycin-treated mice, dmPGE(2) co-treated mice exhibited a reduced degree of body weight loss and mortality rate as well as of lung damage and inflammation, as shown by the significant reduction of: (1) lung infiltration by leukocytes; (2) myeloperoxidase activity; (3) IL-1, TNF-alpha, and nitrotyrosine immunostaining; (4) lung edema; and (5) histologic evidence of lung injury and collagen deposition. In a separate set of experiments, dmPGE(2) treatment was started 3 days after bleomycin administration, and the evaluation of lung damage and inflammation was assessed 4 days later. Importantly, delayed administration of dmPGE(2) also was able to protect from inflammation and lung injury induced by bleomycin. These results, indicating that dmPGE(2) is able to prevent and to reduce bleomycin-induced lung injury through its regulatory and anti-inflammatory properties, encourage further research to find new options for the treatment of pulmonary fibrosis.

TGF-beta1 targets the GSK-3beta/beta-catenin pathway via ERK activation in the transition of human lung fibroblasts into myofibroblasts.

Transforming growth factor-beta1 (TGF-beta1) is known to induce the transition of human lung fibroblasts to myofibroblasts, a primary event in the pathogenesis of idiopathic pulmonary fibrosis. The molecular pathways involved in myofibroblast transformation are only partially identified. We found that a 24-h treatment with TGF-beta1 (10 ng/ml) induced alpha-smooth actin (SMA) expression and collagen production in human lung fibroblasts. These effects were abrogated by PD98059, a specific inhibitor of the mitogen-activated protein kinase (MAPK) pathway. TGF-beta1 treatment activated the MAPK pathway, as shown by an increased phosphorylation of extracellular-regulated kinases (ERK)1/2 after 30 min of exposure. TGF-beta1 also increased the expression of the Ser-9-phosphorylated inactive form of glycogen synthase kinase-3beta (GSK-3beta), an effect that was largely attenuated by PD98059. A nuclear translocation of beta-catenin in human lung fibroblasts was observed 2h after TGF-beta1 addition both by confocal microscopy and nuclear protein analysis. At this time, TGF-beta1 also increased the total levels of beta-catenin, an effect that was prevented by PD98059. Similarly to TGF-beta1, the GSK-3beta inhibitor lithium chloride (10mM), increased the total levels of beta-catenin and promoted alpha-SMA expression and collagen production. This study demonstrates that TGF-beta1 induces alpha-SMA expression and collagen production in human lung fibroblasts via ERK1/2 activation, GSK-3beta inhibition and nuclear beta-catenin translocation. The evidence that the silencing of beta-catenin by siRNAs was able to prevent the induction of alpha-SMA expression in TGF-beta1-treated fibroblasts further supports the hypothesis of a contribution of the GSK-3beta/beta-catenin pathway in the pathogenesis of idiopathic pulmonary fibrosis.

Activation of cytosolic phospholipase A2 and 15-lipoxygenase by oxidized low-density lipoproteins in cultured human lung fibroblasts.

In cell cultures of human lung fibroblasts, we found that oxidized LDL (oxLDL), after 24-h treatment, stimulated arachidonic acid release. A putative role for phospholipases A(2) and MAPK activities in this process was postulated. Consequently, we studied the contribution of either Ca(2+)-dependent, cytosolic phospholipase A(2) (cPLA(2)) or Ca(2+)-independent phospholipase A(2) (iPLA(2)), and the role of the MAP kinase family in oxLDL toxicity to fibroblastic cells in vitro. Activation of extracellular signal-regulated kinases ERK1/2, p38 and c-Jun NH(2)-terminal kinase (JNK) was also assessed with Western blotting. Compared with cellular samples untreated or treated with native LDL, treatment with oxLDL (50-100 microM hydroperoxides) for 24 h significantly increased the levels of either cPLA(2) protein expression or constitutively phosphorylated cPLA(2) protein; in addition we observed enzyme translocation to membranes. iPLA(2) activity was not stimulated by oxLDL. Arachidonic acid release appeared to be associated with phosphorylation of ERK1/2 which was significantly enhanced in a dose-dependent manner whereas no activation of p38 and JNKs was found, indicating that these MAPKs are not involved in mediating the maximal oxLDL response. Western blotting on subcellular fractions and confocal microscopy analyses confirmed an increase in 15-lipoxygenase (15-LO) protein expression and translocation upon activation. A significant increase of cyclooxygenase-2 expression into membrane fraction was also found. Collectively, the data presented link the stimulation of ERK-cPLA(2)-15-LO pathway by oxLDL to the prooxidant mechanism of the lipoprotein complex. It may initially stimulate the fibroblast reaction against the oxidation challenge as well as metabolic repair, such as during lung inflammation and pulmonary fibrosis.

Protective effect of orally administered carnosine on bleomycin-induced lung injury.

Carnosine is an endogenously synthesized dipeptide composed of beta-alanine and L-histidine. It acts as a free radical scavenger and possesses antioxidant properties. Carnosine reduces proinflammatory and profibrotic cytokines such as transforming growth factor-beta (TGF-beta), IL-1, and TNF-alpha in different experimental settings. In the present study, we investigated the efficacy of carnosine on the animal model of bleomycin-induced lung injury. Mice were subjected to intratracheal administration of bleomycin and were assigned to receive carnosine daily by an oral bolus of 150 mg/kg. One week after fibrosis induction, bronchoalveolar lavage (BAL) cell counts and TGF-beta levels, lung histology, and immunohistochemical analyses for myeloperoxidase, TGF-beta, inducible nitric oxide synthase, nitrotyrosine, and poly(ADP-ribose) polymerase were performed. Finally, apoptosis was quantified by terminal deoxynucleotidyltransferase-mediated UTP end-labeling assay. After bleomycin administration, carnosine-treated mice exhibited a reduced degree of lung damage and inflammation compared with wild-type mice, as shown by the reduction of 1) body weight, 2) mortality rate, 3) lung infiltration by neutrophils (myeloperoxidase activity and BAL total and differential cell counts), 4) lung edema, 5) histological evidence of lung injury and collagen deposition, 6) lung myeloperoxidase, TGF-beta, inducible nitric oxide synthase, nitrotyrosine, and poly(ADP-ribose) polymerase immunostaining, 7) BAL TGF-beta levels, and 8) apoptosis. Our results indicate that orally administered carnosine is able to prevent bleomycin-induced lung injury likely through its direct antioxidant properties. Carnosine is already available for human use. It might prove useful as an add-on therapy for the treatment of fibrotic disorders of the lung where oxidative stress plays a role, such as for idiopathic pulmonary fibrosis, a disease that still represents a major challenge to medical treatment.

Bradykinin differentiates human lung fibroblasts to a myofibroblast phenotype via the B2 receptor.

BACKGROUND: The identification of factors mediating the transition of lung fibroblasts into myofibroblasts is considered fundamental in the comprehension of abnormal reparative processes. Bradykinin, a mediator known for its proinflammatory action, is able to induce cytokine production and contractility in fibroblast cultures. OBJECTIVES: In this study the ability of bradykinin to drive fibroblast into a myofibroblast phenotype at the cellular and molecular level was evaluated. METHODS: alpha-Smooth muscle actin (alpha-SMA) expression and TGF-beta in bradykinin stimulated fibroblasts were tested by means of flow cytometry, Western blot, and RT-PCR. Cell proliferation and collagen production were evaluated by the colorimetric methylthiazol tetrazolium assay and sirius red assay, respectively. Which bradykinin receptor mediates the expression of alpha-SMA was evaluated using selective B1 and B2 blocking agents. Furthermore, the effect of bradykinin on extracellular signal-regulated kinase 1/2 phosphorylation was explored. RESULTS: Bradykinin caused in lung fibroblasts a significant increase in alpha-SMA at the cellular and molecular level. The B2 receptor was held responsible for this effect because a specific receptor antagonist had entirely blocked this effect. Bradykinin was able to induce fibroblast proliferation and collagen production. Bradykinin significantly activated mitogen-activated protein kinase pathway by phosphorylating extracellular signal-regulated kinase 1/2, whereas PD98059, a specific inhibitor, was able to block myofibroblast induction. Although bradykinin induced an increase of TGF-beta on fibroblasts, the blockage of this cytokine did not alter alpha-SMA expression. CONCLUSION: The data support the hypothesis that bradykinin may be involved in bronchial remodeling and lung fibrosis beyond its well recognized proinflammatory activity, also suggesting a new potential therapeutic strategy to control altered reparatory processes.

Interaction between human lung fibroblasts and T-lymphocytes prevents activation of CD4+ cells.

BACKGROUND: T lymphocytes are demonstrated to play an important role in several chronic pulmonary inflammatory diseases. In this study we provide evidence that human lung fibroblasts are capable of mutually interacting with T-lymphocytes leading to functionally significant responses by T-cells and fibroblasts. METHODS: Human lung fibroblast were co-cultured with PMA-ionomycin activated T-CD4 lymphocytes for 36 hours. Surface as well as intracellular proteins expression, relevant to fibroblasts and lymphocytes activation, were evaluated by means of flow cytometry and RT-PCR. Proliferative responses of T lymphocytes to concanavalin A were evaluated by the MTT assay. RESULTS: In lung fibroblasts, activated lymphocytes promote an increase of expression of cyclooxygenase-2 and ICAM-1, expressed as mean fluorescence intensity (MFI), from 5.4 +/- 0.9 and 0.7 +/- 0.15 to 9.1 +/- 1.5 and 38.6 +/- 7.8, respectively. Fibroblasts, in turn, induce a significant reduction of transcription and protein expression of CD69, LFA-1 and CD28 in activated lymphocytes and CD3 in resting lymphocytes. In activated T lymphocytes, LFA-1, CD28 and CD69 expression was 16.6 +/- 0.7, 18.9 +/- 1.9 and 6.6 +/- 1.3, respectively, and was significantly reduced by fibroblasts to 9.4 +/- 0.7, 9.4 +/- 1.4 and 3.5 +/- 1.0. CD3 expression in resting lymphocytes was 11.9 +/- 1.4 and was significantly reduced by fibroblasts to 6.4 +/- 1.1. Intracellular cytokines, TNF-alpha and IL-10, were evaluated in T lymphocytes. Co-incubation with fibroblasts reduced the number of TNF-alpha positive lymphocytes from 54.4% +/- 6.12 to 30.8 +/- 2.8, while IL-10 positive cells were unaffected. Finally, co-culture with fibroblasts significantly reduced Con A proliferative response of T lymphocytes, measured as MTT absorbance, from 0.24 +/- 0.02 nm to 0.16 +/- 0.02 nm. Interestingly, while the activation of fibroblasts is mediated by a soluble factor, a cognate interaction ICAM-1 mediated was demonstrated to be responsible for the modulation of LFA-1, CD28 and CD69. CONCLUSION: Findings from this study suggest that fibroblasts play a role in the local regulation of the immune response, being able to modulate effector functions of cells recruited into sites of inflammation.