Clinical Practice Guideline for the prevention and early detection of breast and ovarian cancer in women from HBOC (hereditary breast and ovarian cancer) families.

An estimated 10% of breast cancer cases exhibit a higher familial incidence, and functional mutations in BRCA (breast cancer-gene) 1 or 2 are responsible for the development of malignant tumors in approximately half of these cases. Women with a germline mutation in either of the two genes have a lifetime risk of up to 85% to develop breast cancer, and of up to 60% risk to develop ovarian cancer. This clinical practice guideline defines the individual and familial tumor constellations that represent an indication for BRCA germline testing. It also describes the therapeutic options (early detection programme vs prophylactic surgery) that arise from the result of a BRCA mutational analysis. This guideline further includes recommendations regarding the use of multigene panels and therapeutic aspects that arise from the selective use of poly ADP ribose polymerase (PARP) inhibitors in patients with known BRCA1 or 2 mutations. It replaces the previous version of the "Clinical Practice Guideline for the Prevention and Early Detection of Breast- and Ovarian Cancer in women from HBOC (hereditary breast and ovarian cancer) families" which was published in 2012.

Clinical implications of genetic testing for BRCA1 and BRCA2 mutations in Austria.

The objective of this study was to describe the experience of genetic testing in Austrian women with a BRCA1 or BRCA2 mutation in terms of preventive measures taken and incident cancers diagnosed. We collected clinical information on 246 Austrian women with a BRCA1 or BRCA2 mutation tested between 1995 and 2012 and followed 182 of them for an average of 6.5 years. Of the 90 women who were cancer-free at baseline, 21.4% underwent preventive bilateral mastectomy, 46.1% had preventive bilateral salpingo-oophorectomy, and 1 took tamoxifen; 58.8% of the at-risk women underwent at least one screening breast magnetic resonance imaging (MRI). Of the 85 women with breast cancer, 69.4% had a unilateral mastectomy or lumpectomy and 30.6% had a contralateral mastectomy. In the follow-up period, 14 new invasive breast cancers (6 first primary and 8 contralateral), 1 ductal carcinoma in situ case, 2 incident ovarian cancer cases, and 1 peritoneal cancer were diagnosed. In Austria, the majority of healthy women with a BRCA1 or BRCA2 mutation opt for preventive oophorectomy and MRI screening to manage their breast cancer risk; few have preventive mastectomy or take tamoxifen.

Presence of intratumoral stem cells in breast cancer patients with or without BRCA germline mutations.

BACKGROUND: BRCA-1/2 germline mutations are responsible for early onset breast cancer and familial association. The underlying causes of the characteristic phenotypic behavior are not completely understood, but mammary stem cells appear to have a key role in this process. MATERIALS AND METHODS: We have investigated the presence of mammary stem / progenitor cells in normal tissues and in tumor tissues obtained from women with and without BRCA1/2 germline mutations by utilizing ALDH-1 immunohistochemistry. RESULTS: Isolated ALDH-1 positive cells were found in 15/28 (54%) of breast cancer samples from women with BRCA 1 or 2 mutations and in 33 /51 (65%) of matched sporadic breast cancer cases (p=0.5949, Chi Square test). While mammary stem cells were also detected in non-malignant breast lesions, only 41% of the tissues contained ALDH-1 positive cells (p=0.0371, Chi Square test). In patients with BRCA germline mutations ALDH-1 positive cells were more common in p53 positive (p=0.0028, Chi Square test) tumors, in high grade (p=0.0796), and in larger tumors (p=0.0604), while no such association was seen in sporadic cancer cases. In our patients, the expression of ALDH-1 positive cells in breast cancer was neither associated with disease-free and overall survival, nor time to metastasis. CONCLUSION: Breast cancers from BRCA mutation carriers do not harbor more ALHD-1 positive cells than sporadic tumors, and their more aggressive phenotype can thus not be explained by an increased stem cell pool. The presence of ALDH-1 in normal breast tissue suggests that additional factors determine the biological behavior of mammary stem cells.

Cardiac tissue ablation with catheter-based microwave heating.

The common condition of atrial fibrillation is often treated by cutting diseased cardiac tissue to disrupt abnormal electrical conduction pathways. Heating abnormal tissue with electromagnetic power provides a minimally invasive surgical alternative to treat these cardiac arrhythmias. Radio frequency ablation has become the method of choice of many physicians. Recently, microwave power has also been shown to have great therapeutic benefit in medical treatment requiring precise heating of biological tissue. Since microwave power tends to be deposited throughout the volume of biological media, microwave heating offers advantages over other heating modalities that tend to heat primarily the contacting surface. It is also possible to heat a deeper volume of tissue with more precise control using microwaves than with purely thermal conduction or RF electrode heating. Microwave Cardiac Ablation (MCA) is used to treat heart tissue that allows abnormal electrical conduction by heating it to the point of inactivation. Microwave antennas that fit within catheter systems can be positioned close to diseased tissue. Specialized antenna designs that unfurl from the catheter within the heart can then radiate specifically shaped fields, which overcome problems such as excessive surface heating at the contact point. The state of the art in MCA is reviewed in this paper and a novel catheter-based unfurling wide aperture antenna is described. This antenna consists of the centre conductor of a coaxial line, shaped into a spiral and insulated from blood and tissue by a non-conductive fluid filled balloon. Initially stretched straight inside a catheter for transluminal guiding, once in place at the cardiac target, the coiled spiral antenna is advanced into the inflated balloon. Power is applied in the range of 50-150 W at the reserved industrial, scientific and medical (ISM) frequency of 915 MHz for 30-90 s to create an irreversible lesion. The antenna is then retracted back into the catheter for removal. Simulated and experimental measurements on phantoms, in vitro animal organ tissue and living animals have shown that these microwave applicators deliver the intended therapeutic lesions that are both wider and deeper than those generated by RF ablation or other recently reported microwave applicators.

New perfluorocarbon system for multilayer growth of anchorage-dependent mammalian cells.

A novel tissue culture system has been developed that supports the multilayer growth of Hep G2 cells. The system consists of growing cells on collagen-coated perfluorocarbon substrata in the wells of a multi-well plate designed so that, even at very high densities, the oxygen in the cultures is replenished as rapidly as it is consumed. Hep G2 cells, which are typically contact inhibited, grow to form more than 10 layers of cells that continue to secrete albumin. Both multilayer growth and high rates of albumin depend on using a very enriched nutrient medium, compared to media usually used for monolayer culture of Hep G2 cells. The role played by increased oxygenation, enriched media, and the unique properties of the perfluorocarbon substrata for the 3-D growth of anchorage-dependent cells is discussed.

Monte Carlo simulations of electromagnetic wave scattering from a random rough surface with three-dimensional penetrable buried object: mine detection application using the steepest-descent fast multipole method.

We present a statistical study of the electric field scattered from a three-dimensional penetrable object buried under a two-dimensional random rough surface. Monte Carlo simulations using the steepest-descent fast multipole method (SDFMM) are conducted to calculate the average and the standard deviation of the near-zone scattered fields. The SDFMM, originally developed at the University of Illinois at Urbana-Champaign, has been modified to calculate the unknown surface currents both on the rough ground and on the buried object that are due to excitation by a tapered Gaussian beam. The rough ground medium used is an experimentally measured typical dry Bosnian soil with 3.8% moisture, while the buried object represents a plastic land mine modeled as an oblate spheroid with dimensions and burial depth smaller than the free-space wavelength. Both vertical and horizontal polarizations for the incident waves are studied. The numerical results show that the TNT mine signature is almost 5% of the total field scattered from the ground. Moreover, relatively recognizable object signatures are observed even when the object is buried under the tail of the incident beam. Interestingly, even for the small surface roughness parameters considered, the standard deviation of the object signature is almost 30% of the signal itself, indicating significant clutter distortion that is due to the roughness of the ground.

Microwave ablation using a spiral antenna design in a porcine thigh muscle preparation: in vivo assessment of temperature profile and lesion geometry.

INTRODUCTION: Theoretical studies have suggested that microwave energy can increase the depth of heating compared with radiofrequency energy. A spiral microwave antenna design may have advantages over previous designs using smaller designs because the resulting power deposition pattern is considerably larger than the catheter diameter. We tested the efficacy of a spiral antenna using microwave energy in a porcine thigh muscle preparation. METHODS AND RESULTS: In five anesthetized pigs, the thigh muscle was exposed and bathed in heparinized bovine blood (36 degrees to 37 degrees C). A helical microwave catheter with a fiberoptic thermometer attached to the distal end was positioned perpendicular to the thigh muscle. The antenna-tissue interface and tissue temperatures at depths of 3.0 and 6.0 mm were measured. A 915-MHz microwave generator delivered energy at one of three power outputs (50, 100, or 150 W) for 60 seconds. Seventy lesions were created: 50 W (n = 23), 100 W (n = 24), and 150 W (n = 23). The mean depths at 50, 100, and 150 W were 4.3 +/- 1.8 mm, 7.2 +/- 1.7 mm, and 9.4 +/- 0.9 mm, respectively. Lesion depth (R = 0.96, P = 0.05), maximum surface dimension (R = 0.99, P = 0.06), and volume (R = 0.99, P = 0.04) were closely correlated to the power applied. CONCLUSION: Power is an important determinant of lesion size using a spiral microwave antenna. A novel, spiral microwave antenna design can create lesions of significant depth that may be applicable for the ablative therapy of ventricular tachycardia.

A 2 1/4-turn spiral antenna for catheter cardiac ablation.

To investigate the delivery of microwave energy by a catheter positioned inside the heart for ablating small abnormal regions producing cardiac arrhythmias, a 2 1/4-turn spiral catheter-based microwave antenna applicator has been developed. The antenna consists of the center conductor with continuous insulating material extending from the coaxial feed cable formed into a spiral antenna. The insulator completely isolates the center conductor from tissue. Phantom experiments were performed on homogeneous tissue equivalent medium. The reflection coefficient of the antenna at different frequencies and for different spiral lengths, the time course and temperature profile of an ablation, and the dosimetry of power versus temperature, all indicate that the high-power heating patterns from this antenna are both wider and deeper than with the other microwave antenna systems and radio-frequency electrodes.

Novel oxygenation system supports multilayer growth of HeLa cells.

We have developed a novel substratum in which gelatin is bonded to a reservoir of perfluorodecalin using a perfluoroalkylating technique. This forms a stable substratum supporting good adhesion for cells. HeLa cells cultured on this substratum continued to grow exponentially after the surface was covered with a monolayer forming a tissue-like structure of more than 19 layers of cells. Histological sectioning and staining of the block of tissue formed revealed the presence of mitotic figures deep within the structure. Every cell was surrounded by other cells, similar to growth of cells in vivo. This technique opens up a new approach to studying problems involved in cell-cell interaction and development of histotypic structures in vitro.

Investigation of AC-DC magnetic field effects in planar phospholipid bilayers.

Observations recently reported by others indicate that a combination of a weak dc magnetic field and extremely-low-frequency ac magnetic field can produce resonant effects in biological systems. We report measurements of the effects of combined dc and ac magnetic fields on the dc current through channel-free planar phospholipid membranes. The combined dc-ac magnetic fields did affect the dc current through planar phospholipid membranes, but not in every membrane, and not consistently at the same values of magnetic flux density and frequency. None of our measurements showed resonant response akin to the cyclotron-like resonance reported in diatoms [Smith et al., 1987] and lymphocytes [Liboff et al., 1987].

Calcium binding to metallochromic dyes and calmodulin in the presence of combined, AC-DC magnetic fields.

The possibility that weak, ac and dc magnetic fields in combination may affect binding equilibria of calcium-ions (Ca2+) was investigated with two metallochromic dyes as calcium-binding molecules: murexide and arsenazo III. Calcium-dye equilibria were followed by measuring solution absorbances with a fiber-optic spectrophotometer. A Ca(2+)-arsenazo solution was also used indirectly to monitor the binding of Ca2+ to calmodulin. Parallel, ac and dc magnetic fields were applied to each preparation. The ac magnetic field was held constant during each of a series of experiments at a frequency in the range between 50 and 120 Hz (sine wave) or at 50 pps (square wave) and at an rms flux density in the range between 65 and 156 microT. The dc magnetic field was then varied from 0 to 299 microT at 1.3 microT increments. The magnetic fields did not measurably affect equilibria in the binding of metallochromic dyes or calmodulin to Ca2+.

Health problems of homeless children in New York City.

We reviewed the outpatient medical records of 265 homeless children less than 5 years of age in New York City and compared them with children of similar low socioeconomic status (SES) attending the same pediatric clinic. The frequency of health problems among the homeless children, including delayed immunizations, elevated blood lead levels, the rate of admission to hospital, and the rate of child abuse and neglect reports, exceeded those for the comparison groups.

An hypothesis on the role of cellular colloid osmotic pressure in determining behavior of cells in vitro including anchorage dependency and maintenance of the differentiated state.

The osmotic problems involved when cells are isolated from tissues are analyzed. Evidence is considered which indicates that in vivo the Na pump is operating at maximal or near maximal rates and that this depends on low leak rates for salts and water due to various aspects of the tissues structure. Dispersion of the tissue results in breakdown of these barriers on free diffusion and the isolated cell is subjected to an enormous increase in passive influx due to colloid osmotic pressure without being able to increase its pumping rate to the extent needed to maintain volume control. It is proposed that the primary problem the cell faces in vitro is to compensate for the effective increase in its colloid pressure, e.g. the colloid osmotic pressure excess, emerging with the breakdown of the tissue structure. The finding that most normal cells have to adhere to a surface in order to grow or "anchorage dependency" is analyzed in terms of the way adhesion and spreading result in changes in ion and water movements into cells enabling them to achieve fluid balance in the face of the colloid pressure excess. It is also proposed that the differentiated state is more dependent on colloid osmotic balance than proliferation. The failure of conditions used in tissue culture to compensate adequately for the colloid pressure excess results in limiting the amount of protein which can be synthesized, dissipation of cellular energy, and changes in orientation of cellular components which contribute directly to the loss of differentiation which occurs during growth in vitro.

Proton-induced uptake of mammalian DNA by dog erythrocyte pink ghosts.

Low pH (below 6) induces the uptake of mammalian DNA in dog erythrocyte pink ghosts. Uptake requires either Ca2+ or Mg2+ and is stimulated by ATP. These agents induce a rapid sphering of the ghosts at 37 degrees C and sphering is required for uptake. Uptake is increased in ghosts which have been incubated 60-90 min before adding the DNA. Uptake is strongly temperature-dependent. Lowering the temperature of a suspension of ghosts taking up DNA at 37-0 degrees C stops uptake. It is concluded that uptake depends on active membrane processes and that it may depend on the capacity of the ghosts to maintain cation exchange.