Incidence Rates of Gynecologic Cancers in the U.S. Active Duty Military Population.

INTRODUCTION: Despite an increasing number of female service members, incidence rates of gynecologic cancers (other than cervical cancer) have not been previously documented in the U.S. active duty military population. This study sought to determine the incidence rates of all gynecologic, including peritoneal, malignancies in the U.S. Active Duty population compared to the general US population as reported in the Surveillance, Epidemiology, and End Results Program database. MATERIALS AND METHODS: Gynecologic cancers diagnosed in U.S. Active Duty women aged 20-59 between 2004 and 2013 were retrospectively ascertained. Cancer cases were identified in both the Automated Central Tumor Registry and the Military Health System Data Repository. All cases in Automated Central Tumor Registry plus cases recorded in Military Health System Data Repository, but not duplicative of Automated Central Tumor Registry cases, were included. Age-specific and age-adjusted incidence rates were calculated in military and Surveillance, Epidemiology, and End Results cases. RESULTS: In U.S. Active Duty women, 327 incident cases of gynecologic cancer were identified. There were 110 cases of cervical cancer, 40 cases of endometrial cancer, 152 cases of ovarian cancer, and 25 other gynecologic malignancies. Of the 327 cases, 154 were ascertained from the Automated Central Tumor Registry database and the remainder from Military Health System Data Repository claims data. The age-adjusted rate of all gynecologic cancers for U.S. Active Duty women was 49.17 per 105 (95%CI 37.58, 65.12), while the age-adjusted rate for Surveillance, Epidemiology, and End Results -18 was 42.09 per 105 (95%CI 41.83, 42.35). The kappa coefficient assessing the overlap between the data sources was -0.1937. Though insufficient in numbers for statistical analysis, the observed proportion of ovarian to cervical cancer cases in active duty women < 45 years of age was substantially greater than in the general population. CONCLUSIONS: U.S. Active Duty women exhibited a similar age-adjusted rate of gynecologic cancer as the general US population. There was suboptimal overlap between the Automated Central Tumor Registry and Military Health System Data Repository databases, indicating the necessity of using both databases in order to obtain reliable data in the active duty population. This study is the current best estimate of a baseline rate of gynecologic cancer in U.S. active duty military women. This rate might change over time as women's roles and exposures in recent and future military conflicts evolve.

Development of new PTK7-targeting aptamer-fluorescent and -radiolabelled probes for evaluation as molecular imaging agents: Lymphoma and melanoma in vivo proof of concept.

Aptamers are single-stranded oligonucleotides that recognize molecular targets with high affinity and specificity. Aptamer that selectively bind to the protein tyrosine kinase-7 (PTK7) receptor, overexpressed on many cancers, has been labelled as probes for molecular imaging of cancer. Two new PTK7-targeting aptamer probes were developed by coupling frameworks from the fluorescent dye AlexaFluor647 or the 6-hydrazinonicotinamide (HYNIC) chelator-labelled to 99mTc. The derivatizations via a 5'-aminohexyl terminal linker were done at room temperature and under mild buffer conditions. Physicochemical and biological controls for both imaging agents were performed verifying the integrity of the aptamer-conjugates by HPLC. Recognition of melanoma (B16F1) and lymphoma (A20) mouse cell lines by the aptamer was studied using cell binding, flow cytometry and confocal microscopy. Finally, in vivo imaging studies in tumour-bearing mice were performed. The new probes were able to bind to melanoma and lymphoma cell lines in vitro, the in vivo imaging in tumour-bearing mice showed different uptake behaviours showing for the fluorescent conjugate good uptake by B cell lymphoma while the radiolabelled conjugate did not display tumour uptake due to its high extravascular distribution, and both showed rapid clearance properties in tumour-bearing mice.

Queer periods: attitudes toward and experiences with menstruation in the masculine of centre and transgender community.

Menstruation has long been viewed as an important aspect of women's health. However, scholars and healthcare providers have only recently begun to recognise that transgender men and people with masculine gender identities also menstruate, thus little is known about their attitudes toward and experiences with menstruation. A sample of masculine of centre and transgender individuals with a mean age of 30 years was recruited online to complete measures of attitudes toward menstruation and menstrual suppression and to answer exploratory questions about their experiences managing menstruation. Participants reported mixed attitudes toward menstruation, but generally positive attitudes toward menstrual suppression. Many participants said that they try to avoid public restrooms during menstruation because of practical and psychological concerns. Implications of our findings for the transgender health are discussed.

Carbohydrate antigens in nipple aspirate fluid predict the presence of atypia and cancer in women requiring diagnostic breast biopsy.

BACKGROUND: The goal of this prospective study was to determine (a) concentrations of the carbohydrate biomarkers Thomsen Friedenreich (TF) antigen and its precursor, Tn antigen, in nipple discharge (ND) collected from women requiring biopsy because of a suspicious breast lesion; and (b) if concentration levels predicted pathologic diagnosis. METHODS: Adult women requiring biopsy to exclude breast cancer were enrolled and ND obtained. The samples from 124 women were analyzed using an anti-TF and anti-Tn monoclonal antibodies in direct immunoassay. RESULTS: The highest median concentration in ND for TF and Tn was in women with ductal carcinoma in situ (DCIS). TF was higher in women with 1) cancer (DCIS or invasive) vs. either no cancer (atypia or benign pathology, p = .048), or benign pathology (p = .018); and 2) abnormal (atypia or cancer) versus benign pathology (p = .016); and was more predictive of atypia or cancer in post- compared to premenopausal women. Tn was not predictive of disease. High TF concentration and age were independent predictors of disease, correctly classifying either cancer or abnormal vs. benign pathology 83% of the time in postmenopausal women. CONCLUSIONS: TF concentrations in ND were higher in women with precancer and cancer compared to women with benign disease, and TF was an independent predictor of breast atypia and cancer. TF may prove useful in early breast cancer detection.

A generalized kinetic model for amine modification of proteins with application to phage display.

Amine modification of filamentous virions (phage particles) is widely used in phage display technology to couple small groups such as biotin or fluorescent dyes to the major coat protein pVIII. We have developed a generalized kinetic model for protein amine modification and applied it to the modification of pVIII with biotin and the near-infrared fluorophor Alexa Fluor 680. Empirically optimized kinetic parameters for the two modification reactions allow the modification level to be predicted for a wide range of virions and modifying reagent concentrations. Virions with 0.03 biotins per pVIII subunit have 50% of the maximal binding capacity for a streptavidin conjugate.

In vivo bacteriophage display for the discovery of novel peptide-based tumor-targeting agents.

A powerful strategy for targeted drug discovery is the use of bacteriophage (phage) display technology for identification of peptide-based tumor targeting agents. Peptide pharmaceuticals may possess clinically desirable properties because of their rapid blood clearance, non-immunogenic nature, and ease of synthesis. Phage display has identified hundreds of different peptide sequences that bind a desired target in vitro. Regrettably, few of these peptides offer good targeting efficacy in vivo. One reason for this is the synthesized peptide may not retain its optimal activity outside the microenvironment of the phage. Another possible explanation is that traditionally, phage selections are performed in vitro outside the complicated milieu of a living animal. Given these shortcomings, we have developed methods to select phage peptide display libraries in living mice, to identify, a priori, phage (and corresponding synthesized peptides) with ideal tumor-targeting propensity.

Phage peptide display.

Molecular imaging is at the forefront in the advancement of in-vivo diagnosis and monitoring of cancer. New peptide-based molecular probes to facilitate cancer detection are rapidly evolving. Peptide-based molecular probes that target apoptosis, angiogenesis, cell signaling and cell adhesion events are in place. Bacteriophage (phage) display technology, a molecular genetic approach to ligand discovery, is commonly employed to identify peptides as tumor-targeting molecules. The peptide itself may perhaps have functional properties that diminish tumor growth or metastasis. More often, a selected peptide is chemically synthesized, coupled to a radiotracer or fluorescent probe, and utilized in the development of new noninvasive molecular imaging probes. A myriad of peptides that bind cancer cells and cancer-associated antigens have been reported from phage library selections. Phage selections have also been performed in live animals to obtain peptides with optimal stability and targeting properties in vivo. To this point, few in-vitro, in-situ, or in-vivo selected peptides have shown success in the molecular imaging of cancer, the notable exception being vascular targeting peptides identified via in-vivo selections. The success of vasculature targeting peptides, such as those with an RGD motif that bind alpha(v)beta(3)integrin, may be due to the abundance and expression patterns of integrins in tumors and supporting vasculature. The discovery of molecular probes that bind tumor-specific antigens has lagged considerably. One promising means to expedite discovery is through the implementation of selected phage themselves as tumor-imaging agents in animals.

Melanoma imaging with pretargeted bivalent bacteriophage.

UNLABELLED: Random bacteriophage (phage) display peptide libraries have traditionally been used for the selection of clones that bind specific tissues, tumors, and antigens. However, once the targeting peptide is synthetically produced, it often displays a lower affinity than the original phage because of a lack of avidity effects and removal from the virion surface. We hypothesized that multivalent bifunctional phage displaying peptides that target novel molecular biomarkers would facilitate the in vivo imaging of cancer. This study provides proof of principle for the use of phage displaying multiple melanocortin-1 receptor-homing peptides for the pretargeting and subsequent imaging of murine melanomas in vivo. METHODS: A 2-step melanoma pretargeting-imaging system was developed by first generating and biotinylating phage that displayed up to 5 copies of alpha-melanocyte-stimulating hormone (alpha-MSH) peptide analogs. Second, streptavidin was conjugated to diethylenetriaminepentaacetic acid for the purpose of radiolabeling with (111)In. RESULTS: The specificity of the MSH2.0 phage for the B16-F1 melanoma was demonstrated both in vitro and in vivo. In vitro micropanning assays with phage at inputs of 10(7) and 10(6) transducing units per milliliter resulted in approximately 200- and approximately 1,000-fold-greater recovery of the MSH2.0 phage over the background, respectively. In vivo distribution studies indicated that melanoma uptake values were 2.6 +/- 1.1, 0.6 +/- 0.2, and 1.0 +/- 0.1 (mean +/- SD) percentage injected dose per gram at 0.5, 6, and 24 h after the injection of (111)In-radiolabeled streptavidin ((111)In-SA). The accumulation of radioactivity within the tumor was 1.8 times greater for the biotinylated MSH2.0 phage than for the biotinylated wild-type phage. These data, combined with reduction by 2.4-fold through competition with a nonradiolabeled alpha-MSH peptide analog, indicated the specific targeting of melanoma tumors in vivo. SPECT/CT image analysis of B16-F1 melanoma-bearing mice showed that intravenously injected biotinylated alpha-MSH phage were retained within melanoma tumors at 4 h after injection of (111)In-SA. CONCLUSION: This study demonstrated the use of multivalent bifunctional phage in a 2-step pretargeting-imaging system.

In vivo selection of phage for the optical imaging of PC-3 human prostate carcinoma in mice.

There is an increasing medical need to detect and spatially localize early and aggressive forms of prostate cancer. Affinity ligands derived from bacteriophage (phage) library screens can be developed to molecularly target prostate cancer with fluorochromes for optical imaging. Toward this goal, we used in vivo phage display and a newly described micropanning assay to select for phage that extravasate and bind human PC-3 prostate carcinoma xenografts in severe combined immune deficiency mice. One resulting phage clone (G1) displaying the peptide sequence IAGLATPGWSHWLAL was fluorescently labeled with the near-infrared fluorophore AlexaFluor 680 and was evaluated both in vitro and in vivo for its ability to bind and target PC-3 prostate carcinomas. The fluorescently labeled phage clone (G1) had a tumor-to-muscle ratio of approximately 30 in experiments. In addition, prostate tumors (PC-3) were readily detectable by optical-imaging methods. These results show proof of principle that disease-specific library-derived fluorescent probes can be rapidly developed for use in the early detection of cancers by optical means.

Remodeling of synaptic structure in sensory cortical areas in vivo.

Although plastic changes are known to occur in developing and adult cortex, it remains unclear whether these changes require remodeling of cortical circuitry whereby synapses are formed and eliminated or whether they rely on changes in the strength of existing synapses. To determine the structural stability of dendritic spines and axon terminals in vivo, we chose two approaches. First, we performed time-lapse two-photon imaging of dendritic spine motility of layer 5 pyramidal neurons in juvenile [postnatal day 28 (P28)] mice in visual, auditory, and somatosensory cortices. We found that there were differences in basal rates of dendritic spine motility of the same neuron type in different cortices, with visual cortex exhibiting the least structural dynamics. Rewiring visual input into the auditory cortex at birth, however, failed to alter dendritic spine motility, suggesting that structural plasticity rates might be intrinsic to the cortical region. Second, we investigated the persistence of both the presynaptic (axon terminals) and postsynaptic (dendritic spine) structures in young adult mice (P40-P61), using chronic in vivo two-photon imaging in different sensory areas. Both terminals and spines were relatively stable, with >80% persisting over a 3 week period in all sensory regions. Axon terminals were more stable than dendritic spines. These data suggest that changes in network function during adult learning and memory might occur through changes in the strength and efficacy of existing synapses as well as some remodeling of connectivity through the loss and gain of synapses.

Acceleration of visually cued conditioned fear through the auditory pathway.

Defensive responses elicited by sensory experiences are critical for survival. Mice acquire a conditioned fear response rapidly to an auditory cue but slowly to a visual cue, a difference in learned behavior that is likely to be mediated by direct projections to the lateral amygdala from the auditory thalamus but mainly indirect ones from the visual thalamus. Here, we show that acquisition of visually cued conditioned fear is accelerated in 'rewired' mice that have retinal projections routed to the auditory thalamus. Visual stimuli induce expression of the immediate early gene Fos (also known as c-fos) in the auditory thalamus and the lateral amygdala in rewired mice, similar to the way auditory stimuli do in control mice. Thus, the rewired auditory pathway conveys visual information and mediates rapid activity-dependent plasticity in central structures that influence learned behavior.

The P2Y2 nucleotide receptor mediates UTP-induced vascular cell adhesion molecule-1 expression in coronary artery endothelial cells.

P2Y2 receptor up-regulation and activation induces intimal hyperplasia and monocyte/macrophage infiltration in the collared rabbit carotid artery model of vascular injury, suggesting a potential role for P2Y2 receptors in monocyte recruitment by vascular endothelium. In this study, we addressed the hypothesis that activation of P2Y2 receptors by extracellular nucleotides modulates the expression of adhesion molecules on vascular endothelial cells that are important for monocyte recruitment. Results indicated that the equipotent P2Y2 receptor agonists UTP or ATP (1-100 microm) stimulated the expression of vascular cell adhesion molecule-1 (VCAM-1) in human coronary artery endothelial cells (HCAEC) in a time- and dose-dependent manner. P2Y2 antisense oligonucleotides inhibited VCAM-1 expression induced by UTP but not by tumor necrosis factor-alpha. Furthermore, UTP induced VCAM-1 expression in human 1321N1 astrocytoma cell transfectants expressing the recombinant P2Y2 receptor, whereas vector-transfected control cells did not respond to UTP. The effect of UTP on VCAM-1 expression in HCAEC was prevented by depletion of intracellular calcium stores with thapsigargin or by inhibition of p38 mitogen-activated protein kinase or Rho kinase, but was not affected by inhibitors of the mitogen-activated protein/extracellular signal-regulated kinase pathway (i.e. MEK1/2). Consistent with a role for VCAM-1 in the recruitment of monocytes, UTP or ATP increased the adherence of monocytic U937 cells to HCAEC, an effect that was inhibited by anti-VCAM-1 antibodies. These findings suggest a novel role for the P2Y2 receptor in the p38- and Rho kinase-dependent expression of VCAM-1 that mediates the recruitment of monocytes by vascular endothelium associated with the development of atherosclerosis.

Chromatic detection and discrimination in the periphery: a postreceptoral loss of color sensitivity.

The peripheral visual field is marked by a deterioration in color sensitivity, sometimes attributed to the random wiring of midget bipolar cells to cone photoreceptors in the peripheral retina (Mullen, 1991; Mullen & Kingdom, 1996). Using psychophysical methods, we explored differences in the sensitivity of peripheral color mechanisms with detection and discrimination of 2-deg spots at 18-deg eccentricity, and find evidence for a postreceptoral locus for the observed loss in sensitivity. As shown before, observers' sensitivity to green was lower than to red in the periphery, although the magnitude of this effect differed across observers. These results suggest that the asymmetry in peripheral sensitivity occurs at a postreceptoral site, possibly a cortical one. In addition, noise masking was used to determine the cone inputs to the peripheral color mechanisms. The masked detection contours indicate that the red and green mechanisms in the periphery respond to the linear difference of approximately equally weighted L- and M-cone contrasts, just as they do in the fovea. Thus, if the midget retinal ganglion system is responsible for red/green color perception in the fovea, it is likely to be responsible at 18-deg eccentricity as well.