Whole-genome and transcriptome analysis enhances precision cancer treatment options.

BACKGROUND: Recent advances are enabling delivery of precision genomic medicine to cancer clinics. While the majority of approaches profile panels of selected genes or hotspot regions, comprehensive data provided by whole-genome and transcriptome sequencing and analysis (WGTA) present an opportunity to align a much larger proportion of patients to therapies. PATIENTS AND METHODS: Samples from 570 patients with advanced or metastatic cancer of diverse types enrolled in the Personalized OncoGenomics (POG) program underwent WGTA. DNA-based data, including mutations, copy number and mutation signatures, were combined with RNA-based data, including gene expression and fusions, to generate comprehensive WGTA profiles. A multidisciplinary molecular tumour board used WGTA profiles to identify and prioritize clinically actionable alterations and inform therapy. Patient responses to WGTA-informed therapies were collected. RESULTS: Clinically actionable targets were identified for 83% of patients, of which 37% of patients received WGTA-informed treatments. RNA expression data were particularly informative, contributing to 67% of WGTA-informed treatments; 25% of treatments were informed by RNA expression alone. Of a total 248 WGTA-informed treatments, 46% resulted in clinical benefit. RNA expression data were comparable to DNA-based mutation and copy number data in aligning to clinically beneficial treatments. Genome signatures also guided therapeutics including platinum, poly-ADP ribose polymerase inhibitors and immunotherapies. Patients accessed WGTA-informed treatments through clinical trials (19%), off-label use (35%) and as standard therapies (46%) including those which would not otherwise have been the next choice of therapy, demonstrating the utility of genomic information to direct use of chemotherapies as well as targeted therapies. CONCLUSIONS: Integrating RNA expression and genome data illuminated treatment options that resulted in 46% of treated patients experiencing positive clinical benefit, supporting the use of comprehensive WGTA profiling in clinical cancer care.

A microfluidic model of the blood-brain barrier to study permeabilization by pulsed electric fields.

Pulsed electric fields interact with the blood-brain barrier (BBB) and have been shown to increase the BBB permeability under some pulsing regimes. Pulsed electric fields may enhance drug delivery to the brain by disrupting the integrity of the BBB and allowing otherwise impermeable drugs to reach target areas. Microfluidic, in vitro models offer an alternative platform for exploring the impact of pulsed electric fields on the BBB because they create physiologically relevant microenvironments and eliminate the confounding variables of animal studies. We developed a microfluidic platform for real-time measurement of BBB permeability pre- and post-treatment with pulsed electric fields. Permeability is measured optically by the diffusion of fluorescent tracers across a monolayer of human cerebral microcapillary endothelial cells (hCMECs) cultured on a permeable membrane. We found that this device is able to capture real-time permeability of hCMEC monolayers for both reversible and irreversible electroporation pulsing regimes. Furthermore, preliminary testing of deep brain stimulation pulsing regimes reveals possible impacts on BBB integrity. This device will enable future studies of pulsed electric field regimes for improved understanding of BBB permeabilization.

The vasculature in chronic adult periodontitis: a qualitative and quantitative study.

Blood vessel features in periodontal pocket soft tissues may be significant in the pathogenesis and progression of chronic periodontitis. The aim of this study was to make a quantitative histological assessment of the vasculature in soft tissue biopsies from patients with chronic adult periodontitis and patients with healthy periodontal tissues. We have also investigated changes in tissue morphology at both the histological and ultrastructural level. Twelve interdental biopsies were obtained, 6 from chronic adult periodontitis patients and 6 from healthy volunteers. The specimens were sliced, fixed in 3% glutaraldehyde, postfixed in 1% buffered osmium tetroxide, dehydrated, and embeded in araldite. One micron semithin sections were differentially stained with a dichromatic technique. The number of blood vessels (BV) for sub-epithelial, superficial and deep connective tissue layers were then assessed. Only in the sub-epithelial connective tissue layer was there a significant increase in the number of blood vessels (95% Confidence interval [CI]) in the chronic adult periodontitis specimens when compared to healthy specimens. The results of this study seem to indicate that a dichromatic staining technique facilitates the identification and quantification of blood vessels in epoxy resin embedded specimens at light microscope level, and that there is an increase in the number of blood vessels in the chronic adult periodontitis lesions.

Can capillaries increase in length without any increase in the number of endothelial cells?

Generally it is believed that one of the causes of vascular expansion is the increase in length of small capillaries, which in turn is attributed to an increase in the number of endothelial cells. In this report we suggest a new hypothesis for a different mechanism for the increased length of small capillaries, which excludes an increase in the number of endothelial cells. This hypothesis is based upon the possibility of a longitudinal expansion of blood vessels, and has been explained and justified through a series of mathematical examples.

Chronic inhalation exposure to ozone and nitric acid elevates stress-inducible heat shock protein 70 in the rat lung.

The ability of urban oxidant and acid air pollutants to induce heat shock proteins (HSPs) in the mammalian lung is not known. Such proteins are known to be correlated with environmental stress and pathophysiological conditions. In this study, stress-inducible HSP 70 was assessed by slot-blotting in rat lungs (N=10 per group) following inhalation exposures for 4 h per day, 3 days per week for 40 weeks to the following pollutants: (a) purified air;(b) 0.15 ppm ozone (O3);(c)50 micrograms/m3 nitric acid (HNO3); or(d) a combination of both 0.15 ppm O3 and 50 micrograms/m3 HNO3. At 24 h following the last exposure, samples from the right apical lobe of the lung were obtained for either slot-blotting or gel electrophoretic separation, subsequent protein immunoblotting, and chemiluminescence detection of HSP 70 levels. Experiments demonstrate that stress-inducible HSP 70 was present constitutively in the control lungs and was separable from the constitutive form of HSP 70. Slot-blotting analysis demonstrate that the O3 and HNO3 exposures alone produced significant elevations of HSP70. Specifically, either O3 or HNO3 alone significantly elevated lung stress-inducible HSP 70 levels by 277% and 221% respectively, above control levels. The group exposed to combined O3 and HNO3 showed a 177% elevation in lung stress-inducible HSP 70 that was significantly greater that the group inhaling purified air, but this effect was less than the effects of either pollutant component alone. Moreover, all exposure groups were significantly different from one another. These results indicate that stress-inducible HSP 70 in the rat lung is highly elevated after chronic inhalation exposures to both O3 and HNO3 when administered either alone or in combination within the range of urban ambient concentrations.

Continuous-flow performance of carbon electrodes modified with immobilized Fe(II)/Fe(III) centers Amperometric response to N(2)O, NO and NO(2).

The amperometric performance of two types of chemically modified carbon electrodes developed for the determination of oxides of nitrogen in continuous-flow systems is presented. The modification consists in immobilization of reversible Fe(II)/Fe(III) centers. The first type of electrode is a simple modification made by direct mixing of carbon paste with tris-4,7-diphenyl-1,10-phenanthroline-iron(II) perchlorate; the other is a glassy-carbon surface modified by oxidative electropolymerization of tris-[5-amino-1,10-phenanthroline]iron(II) perchlorate. Detection is accomplished by transporting an injected sample plug to the sensing surface with the aid of gravitational flow of an aqueous solution of supporting electrolyte. The polymer-coated electrochemical detector compares favourably with the chemically modified carbon paste. It offers excellent resistance to poisoning and a competitive limit of detection [about 2 ppb (2 parts in 10(9)) v/v], at + 1.0 V vs. Ag/AgCl, and good selectivity for NO(2) when used in a thin-layer cell. Incorporation of the cell in a continuous-flow system allows injection of about 120 samples per hour. The typical concentration range amenable to determination is 2-25 ppb v/v but depends on the thickness of the polymeric film. Nitrogen monoxide can also be detected but only in undiluted, pure form. Dinitrogen oxide gives no amperometric signal at any of the modified surfaces.

"Placental polyp": light microscopic and immunohistochemical observations.

A case of the chronic type of placental polyp, occurring in a 37-year-old woman approximately 9 years after abortion of her last known pregnancy, is reported. The placental polyp was predominantly composed of necrotic and hyalinized chorionic villi without identifiable lining trophoblast; however, some villi showed a thin rim of apparently viable syncytiotrophoblast that exhibited focal strong positivity for human chorionic gonadotropin by immunohistochemical studies. Intermediate trophoblast, especially abundant within the intervillous fibrin, appeared most viable and showed strong positivity for human placental lactogen (hPL); syncytiotrophoblast also showed focal positivity for hPL. The basal aspect of the polyp was composed of abundant decidua that contained dilated and ectatic blood vessels. This study demonstrates the presence of cytoplasmic markers for pregnancy in a chronic type of placental polyp, apparently of 9 years' duration, and draws attention to an entity that may be encountered more frequently due to the current prevalence of induced abortions.

Preconcentration and voltammetric measurement of mercury with a crown-ether modified carbon-paste electrode.

Carbon-paste electrodes modified with crown-ethers were constructed by mixing the crown-ethers into a graphite powder/Nujol oil matrix. The electrodes so formed were able to bind mercuric ions chemically, and gave greater voltammetric response to mercury than that of ordinary carbon-paste electrodes. The response was characterized with respect to paste composition, crown-ether, preconcentration period, mercury concentration, reproducibility, possible interferences, and other variables. Best results were obtained with 18-crown-6 and an acetate buffer (pH 4.0). The electrode gave good linearity for 1 x 10(-5)-6 x 10(-6)M mercury, a detection limit of 2 x 10(-6)M, and a relative standard deviation of 11%. The investigation may lead to a new class of modified (complexing) electrodes, with different patterns of reactivity.

An inulin-like clearance based on the atomic absorbance of a copper cluster compound.

Copper cluster ions which contain 14 copper atoms and 12 penicillamine ligands produce no indications of toxicity when injected intravenously in doses of 12 mg/kg (as the hydrated sodium salt). These ions pass freely through the Bowman's capsules and are eliminated with the urinary effluent. The blood plasma and blood serum kinetics of the process may be followed with selectivity and sensitivity using copper's flame atomic absorbance at 324.7 nm, and a first order rate law with a half life of approximately ten minutes is observed. Clearance obtained by this method is inulin-like. The copper cluster ion does not readily permeate the erythrocyte membrane, and its distribution immediately following intravenous injection appears to be essentially that of the plasma volume. Since serum or plasma preparation for atomic absorbance measurements is minimal, the method might have applications in rapid determinations of renal clearance.

Major gynecologic and obstetric surgery in Jehovah's Witnesses.

A retrospective study of 165 Jehovah's Witnesses and 164 Jehovah's Witnesses and 164 control patients compared the morbidity and mortality associated with major obstetric and gynecologic surgery in the 2 groups. There were no deaths and few complications in either group. There were few differences in preoperative and postoperative hemoglobin values. Medicolegal implications of performing major surgery without blood transfusions are discussed. The study adds evidence that major operative procedures can be carried out on Jehovah's Witness patients without blood transfusions or blood products.

Gonadoblastoma with a 45,XO karyotype.

An otherwise asymptomatic 17-year-old phenotypic female examined because of primary amenorrhea and hypoplastic reproductive organs was found to have a 45,XO karyotype. She had some of the stigmata of Turner syndrome. Laparoscopy revealed streak ovaries and an infantile uterus. Ovarian biopsy contained gonadoblastoma that was treated by total abdominal hysterectomy and bilateral salpingo-oophorectomy. Rationale for laparoscopy and ovarian biopsy of this asymptomatic patient and the serendipitous discovery of a malignant neoplasm are discussed.