Evidence of cancer therapy-induced chronic inflammation in the ovary across multiple species: A potential cause of persistent tissue damage and follicle depletion.

Chemotherapy and radiation treatments are known for deleterious effects on the ovary, which can result in prolonged recovery time before ovarian function resumes, including follicular growth after completion of these therapies. To better understand the protracted ovarian dysfunctions after chemotherapy and radiotherapy, we designed a comprehensive study to investigate the underlying mechanisms involved in chronic ovarian damage that prevent follicular development and/or to induce persistent follicle loss. Blood and ovarian samples were collected from reproductive age women, rhesus macaques, and mice after completion of chemotherapy and/or radiotherapy and from age-matched patients and animals without chemotherapy agent or radiation exposure to serve as controls. Serum levels of anti-Müllerian hormone and proinflammatory cytokines, monocyte chemoattractant protein 1 and IL6, were measured. Ovarian tissue was assessed for histopathology and inflammatory cell infiltration, e.g., macrophages and neutrophils, by immuohistochemistry. Serum anti-Müllerian hormone concentrations were lower, whereas proinflammatory cytokine concentrations were higher, in patients and rhesus macaques at ~1 year post-chemotherapy agent and/or radiation exposure compared with controls. The number of primordial follicles reduced in the mouse ovary > 5 weeks after a single injection of cyclophosphamide. Macrophage infiltration was observed in the ovarian cortex of humans and animals. These data suggest that chronic inflammation induced by chemotherapy agents and/or radiation treatment may be associated with persistent ovarian tissue damage, follicle depletion, and functional decline. Interventions that dampen the overactivated inflammatory response may further protect the ovary after completion of chemotherapy and radiotherapy to maintain follicle viability and support continued follicular development in female patients.

Association between vitamin D and ovarian cancer development in BRCA1 mutation carriers.

OBJECTIVE: Women with inherited mutations in BRCA1 gene have a high (40-70%) genetic risk of developing ovarian cancer. Epidemiological studies suggest an inverse correlation between serum vitamin D (VD) levels and the risk of ovarian cancer, but there is a lack of data from BRCA1 mutation (BRCA1 mut) carriers. Therefore, we investigated VD levels and actions in cancer free women with BRCA1 mutations. MATERIALS AND METHODS: Blood, ovary and fallopian tube samples were collected from healthy pre-menopausal women with BRCA1 mut and without BRCA1 mutations (BRCA wt). Serum calcifediol (major circulating form of VD) concentrations were measured by electrochemiluminescence immunoassay. Immunohistochemistry was performed on paraffin-embedded ovarian and fallopian tube sections to determine vitamin D receptor (VDR) expression. Ovarian surface epithelial cells (OSEs) from BRCA1 mut carriers were cultured with or without calcitriol supplementation for 72 hrs. VDR protein levels, cell proliferation and cell viability were analyzed. RESULTS: BRCA1 mut women had lower serum calcifediol levels compared to BRCA wt women (p = 0.003). VDR protein expression was evident in ovarian and the fallopian tube epithelium of BRCA wt patients, but was reduced in BRCA1 mut women. Calcitriol (biologically active VD) supplementation elevated VDR expression in cultured BRCA1 mut OSEs (p = 0.005) and decreased cell proliferation rates in a dose-dependent manner without inducing apoptosis. CONCLUSIONS: VD biosynthesis and signaling via VDR in the ovarian and fallopian tube epithelium are impaired in BRCA1 mut women. VD treatment may limit BRCA1 mut epithelial cell proliferation without affecting cell viability, providing a rationale for exploring the potential for VD in ovarian cancer prevention in BRCA1 mut carriers.

Subcellular localization of FANCD2 is associated with survival in ovarian carcinoma.

OBJECTIVE: Ovarian cancer is a leading cause of death from gynecological cancers. Late diagnosis and resistance to therapy results in mortality and effective screening is required for early diagnosis and better treatments. Expression of the Fanconi Anemia complementation group D2 protein (FANCD2) is reduced in ovarian surface epithelial cells (OSE) in patients with ovarian cancer. FANCD2 has been studied for its role in DNA repair; however multiple studies have suggested that FANCD2 has a role outside the nucleus. We sought to determine whether subcellular localization of FANCD2 correlates with patient outcome in ovarian cancer. METHODS: We examined the subcellular localization of FANCD2 in primary OSE cells from consenting patients with ovarian cancer or a normal ovary. Ovarian tissue microarray was stained with anti-FANCD2 antibody by immunohistochemistry and the correlation of FANCD2 localization with patient outcomes was assessed. FANCD2 binding partners were identified by immunoprecipitation of cytoplasmic FANCD2. RESULTS: Nuclear and cytoplasmic localization of FANCD2 was observed in OSEs from both normal and ovarian cancer patients. Patients with cytoplasmic localization of FANCD2 (cFANCD2) experienced significantly longer median survival time (50 months), versus patients without cytoplasmic localization of FANCD2 (38 months; p < 0.05). Cytoplasmic FANCD2 was found to bind proteins involved in the innate immune system, cellular response to heat stress, amyloid fiber formation and estrogen mediated signaling. CONCLUSIONS: Our results suggest that the presence of cytoplasmic FANCD2 modulates FANCD2 activity resulting in better survival outcome in ovarian cancer patients.

Rapid actions of anti-Müllerian hormone in regulating synaptic transmission and long-term synaptic plasticity in the hippocampus.

Anti-Müllerian hormone (Amh) is a peptide factor that is known to regulate sexual differentiation and gonadal function in mammals. Although Amh is also suggested to be associated with cognitive development and function in the postnatal brain, little is known about its expression or direct effects on neuronal activities in the hippocampus. Therefore, we assessed Amh and its receptor expression in the hippocampus of male and female mice using PCR, Western blot, and immunofluorescence staining. While Amh-specific receptor expression was comparable between males and females, mRNA and protein levels of Amh were higher in females than those of males. Electrophysiological recordings on acute hippocampal slices showed that exogenous Amh protein addition increased synaptic transmission and long-term synaptic plasticity at the Cornu Ammonis (CA) 3-CA1 synapses. Amh exposure also increased the excitatory postsynaptic potential at CA1 synapses. Our findings support direct and rapid actions of Amh as a paracrine and/or autocrine factor in regulating hippocampal neuronal activities. Data provide functional evidence of Amh-mediated postsynaptic modulation of synaptic transmission and Amh-regulated long-term synaptic plasticity in the hippocampus. These results suggest a potential role of Amh in learning and memory, and a possible cause of the sex differences in cognitive development and function.

Stage-dependent actions of antimüllerian hormone in regulating granulosa cell proliferation and follicular function in the primate ovary.

OBJECTIVE: To study the direct action and physiological role of antimüllerian hormone (AMH) in regulating ovarian follicular development and function in vivo in primates. DESIGN: Animals were assigned to six treatment sequences in a crossover design study. Intraovarian infusion was performed during the follicular phase of the menstrual cycle with agents including: control vehicle; recombinant human AMH (rhAMH); and neutralizing anti-human AMH antibody (AMHAb). Before ovariectomy after the final treatment, the animals received intravenous injections of bromodeoxyuridine (BrdU). SETTING: National primate research center. ANIMALS: Adult female rhesus macaques (Macaca mulatta). INTERVENTIONS: None. MAIN OUTCOME MEASURES: Cycle length, follicle cohorts, and serum steroid levels were assessed. Ovarian histology, as well as granulosa cell (GC) proliferation and oocyte viability, were evaluated. RESULTS: In vehicle-infused ovaries, a dominant follicle was observed at midcycle E2 peak. However, rhAMH-treated ovaries exhibited an increased number of small antral follicles, whereas AMHAb-treated ovaries developed multiple large antral follicles. Serum E2 levels in the follicular phase decreased after rhAMH infusion and increased after AMHAb infusion. The rhAMH infusion increased serum T levels. Whereas early-growing follicles of rhAMH-treated ovaries contained BrdU-positive GCs, antral follicles containing BrdU-positive GCs were identified in AMHAb-treated ovaries. Autophagy was observed in oocytes of early-growing and antral follicles exposed to AMHAb and rhAMH, respectively. CONCLUSIONS: AMH enhanced early-stage follicle growth, but prevented antral follicle development and function via its stage-dependent regulation of GC proliferation and oocyte viability. This study provides information relevant to the pathophysiology of ovarian dysfunction and the treatment of infertility.

Comparative analysis of graded blends of palm kernel oil, palm kernel stearin and palm stearin.

A comprehensive analysis of graded binary and ternary blends of palm kernel oil (PKO), palm kernel stearin (PKS) and palm stearin (POs) was conducted, including solid fat content, iso-solid diagram, thermal properties, polymorphism and microstructure. Both PKO/POs and PKS/POs blends showed eutectic behavior. While PKO/PKS blends displayed good compatibility and only slightly monotectic effect at high temperature (above 30 °C). POs addition elevated binary phase transition temperature and accelerated the ß crystal formation. PKO/POs and PKS/POs binaries showed ß' polymorphism, ß and ß' polymorphism, and ß polymorphism with different POs additions, while all of PKO/PKS blends showed ß' polymorphism. Ternary blends (PKO/PKS/POs) showed eutectic effects, especially at high temperature (25 and 30 °C). POs addition made ternary blends undergo a polymorphism transformation from ß' to ß. Our findings open up the possibility of non-hydrogenated fat products as well as develop specific food formulations.

Novel Method for Assessment of His Bundle Pacing Morphology Using Near Field and Far Field Device Electrograms.

BACKGROUND: The 12-lead ECG is considered the gold standard to differentiate between selective (S), nonselective (NS) His bundle pacing (HBP), and right ventricular septal capture in routine clinical practice. We sought to assess the utility of device EGM recordings as a tool to identify the type of HBP morphology. METHODS: One hundred forty-eight consecutive patients underwent HBP with a 3830 Select Secure lead (Medtronic, Inc) at 3 centers between October 2016 and October 2017. The near field V-EGM morphology (NF EGM), near field V-EGM time to peak (NFTime to peak), and far-field EGM QRS duration (QRSd) were recorded while pacing the His lead with simultaneous 12-lead ECG rhythm strips. RESULTS: Indications for HBP were sinus node dysfunction, atrioventricular conduction disease, and cardiac resynchronization therapy in 68 (46%), 56 (38%), and 24 (16%) patients, respectively. Baseline QRSd was 108±38 ms with QRSd >120 ms in 57 (39%) patients (27 right bundle branch block, 18 left bundle branch block, and 12 intraventricular conduction delay). S-HBP was noted in 54 (36%) patients. A positive NFEGM and NFTime to peak >40 ms were highly sensitive (94% and 93%, respectively) and specific (90% and 94%) for S-HBP irrespective of baseline QRSd. All 3 parameters (+NFEGM, NFTime to peak >40 ms, and far-field EGM QRSd <120 ms) had high negative predictive value (97%, 95%, and 92%). A novel device-based algorithm for S-HBP was proposed. EGM transitions correlated with ECG transitions during threshold testing and can help accurately differentiate between S-HBP, NS-HBP, and right ventricular septal pacing with a cumulative positive predictive value of 91% (positive predictive value =100% in patients with baseline QRSd <120 ms). CONCLUSIONS: We propose a novel and simple criteria for accurate differentiation between S-HBP, NS-HBP, and right ventricular septal capture morphologies by careful analysis of device EGMs alone. This study paves the way for future studies to assess autocapture algorithms for devices with HBP.

Visualized phase behavior of binary blends of coconut oil and palm stearin.

The graded blends of coconut oil (CNO) and palm stearin (POs) phase behavior was studied in the present work, by using pulsed nuclear magnetic resonance (p-NMR), differential scanning calorimetry (DSC) and X-ray scattering (XRD). The kinetic phase diagram which was fitting to DSC data by polynomial equation (R > 0.95), indicated that the CNO-POs binary blends displayed monotectic behavior. The CNO-POs binary system displayed immiscible solid structures (ß and ß' polymorphism) with the addition of POs in the range of 10-60%, beyond which it showed miscible solid structures (ß polymorphism), respectively. Moreover, the presence of POs could elevate the liquid phase transition temperature and transform ß' polymorph into ß. These variations in phase behavior were reflected in the morphology of the binary blends. Our findings not only broaden the application of CNO and POs with novel attributes, but also direct the production of high quality non-hydrogenated fat-containing products.

Influence of indigenous minor components on fat crystal network of fully hydrogenated palm kernel oil and fully hydrogenated coconut oil.

Purification of triglycerides from fully hydrogenated palm kernel oil (FHPKO) and fully hydrogenated coconut oil (FHCNO) was performed by a chromatographic method. Lipid composition, thermal properties, polymorphism, isothermal crystallization behaviour, nanostructure and microstructure of FHPKO, FHPKO-triacylglycerol (TAG), FHCNO and FHCNO-TAG were evaluated. Removal of minor components had no effect on triglycerides composition. However, the presence of the minor components did increase the slip melting point and promote onset of crystallization. Furthermore, the thickness of the nanoscale crystals increased, and polymorphic transformation from ß' to ß occurred in FHPKO after the removal of minor components, and from α to ß' in FHCNO. Sharp changes in the values of the Avrami constant K and exponent n suggested that the presence of minor components changed the crystal growth mechanism. The PLM results indicated that a coarser crystal structure with lower fractal dimension appeared after the removal of minor components from both FHPKO and FHCNO.

Non-triglyceride components modulate the fat crystal network of palm kernel oil and coconut oil.

PKO and CNO are composed of 97-98% triacylglycerols and 2-3% minor non-triglyceride components (FFA, DAG and MAG). Triglycerides were separated from minor components by chromatographic method. The lipid composition, thermal properties, polymorphism, isothermal crystallization behavior, nanostructure and microstructure of PKO, PKO-TAG, CNO and CNO-TAG were evaluated. Removal of minor components had no effect on lipid composition and equilibrium solid fat contents. However, presence of minor components did increase the slip melting point and promoted the onset of crystallization from DSC crystallization profiles. The thickness of the nanoscale crystals increased with no polymorphic transformation after removing the minor components. Crystallization kinetics revealed that minor components decreased crystal growth rate with higher t1/2. Sharp changes in the values of the Avrami constant k and exponent n were observed for all fats around 10°C. Increases in n around 10°C indicated a change from one-dimensional to multi-dimensional growth. From the results of polarized light micrographs, the transformation from the coarser crystal structure to tiny crystal structure occurred in microstructure networks at the action of minor components.

No Evidence That Genetic Variation in the Myeloid-Derived Suppressor Cell Pathway Influences Ovarian Cancer Survival.

Background: The precise mechanism by which the immune system is adversely affected in cancer patients remains poorly understood, but the accumulation of immunosuppressive/protumorigenic myeloid-derived suppressor cells (MDSCs) is thought to be a prominent mechanism contributing to immunologic tolerance of malignant cells in epithelial ovarian cancer (EOC). To this end, we hypothesized genetic variation in MDSC pathway genes would be associated with survival after EOC diagnoses.Methods: We measured the hazard of death due to EOC within 10 years of diagnosis, overall and by invasive subtype, attributable to SNPs in 24 genes relevant in the MDSC pathway in 10,751 women diagnosed with invasive EOC. Versatile Gene-based Association Study and the admixture likelihood method were used to test gene and pathway associations with survival.Results: We did not identify individual SNPs that were significantly associated with survival after correction for multiple testing (P < 3.5 × 10-5), nor did we identify significant associations between the MDSC pathway overall, or the 24 individual genes and EOC survival.Conclusions: In this well-powered analysis, we observed no evidence that inherited variations in MDSC-associated SNPs, individual genes, or the collective genetic pathway contributed to EOC survival outcomes.Impact: Common inherited variation in genes relevant to MDSCs was not associated with survival in women diagnosed with invasive EOC. Cancer Epidemiol Biomarkers Prev; 26(3); 420-4. ©2016 AACR.

Dispersion behavior and aqueous foams in mixtures of a vesicle-forming surfactant and edible nanoparticles.

In an attempt to prepare ultrastable aqueous foams composed entirely of food-grade ingredients, we describe the foamability and foam stability of aqueous phases containing either calcium carbonate particles (CaCO3), sodium stearoyl lactylate surfactant (SSL), or their mixtures. Techniques including zeta potential measurements, adsorption isotherm determination, contact angles and optical and cryo-scanning electron microscopy are used to probe the interaction between particles and surfactant molecules. Aqueous dispersions of inherently hydrophilic cationic CaCO3 nanoparticles do not foam to any great extent. By contrast, aqueous dispersions of anionic SSL, which forms a lamellar phase/vesicles, foam progressively on increasing the concentration. Despite their foamability being low compared to that of micelle-forming surfactant sodium dodecyl sulfate, they are much more stable to collapse with half-lives (of up to 40 days) of around 2 orders of magnitude higher above the respective aggregation concentrations. We believe that, in addition to surfactant lamellae around bubbles, the bilayers within vesicles contain surfactant chains in a solidlike state yielding indestructible aggregates that jam the aqueous films between bubbles, reducing the drainage rate and both bubble coalescence and gas-transfer between bubbles. In mixtures of particles and surfactant, the adsorption of SSL monomers occurs on particle surfaces, leading to an increase in their hydrophobicity, promoting particle adsorption to bubble surfaces. Ultrastable foams result with half-lives of around an order of magnitude higher again at low concentrations and foams which lose only around 30% of their volume within a year at high concentrations. In the latter case, we evidence a high surface density of discrete surfactant-coated particles at bubble surfaces, rendering them stable to coalescence and disproportionation.