Long-term progestin contraceptives (LTPOC) induce aberrant angiogenesis, oxidative stress and apoptosis in the guinea pig uterus: A model for abnormal uterine bleeding in humans

BACKGROUND: Irregular uterine bleeding is the major side effect of, and cause for, discontinuation of long-term progestin-only contraceptives (LTPOCs). The endometria of LTPOC-treated women display abnormally enlarged, fragile blood vessels (BV), decreased endometrial blood flow and oxidative stress. However, obtaining sufficient, good quality tissues have precluded elucidation of the mechanisms underlying these morphological and functional vascular changes. METHODS: The current study assessed the suitability of the guinea pig (GP) as a model for evaluating the uterine effects of LTPOC administration. Thus GPs were treated with a transdermal pellet for 21 days and examined for endometrial histology, angiogenic markers as well as markers of oxidative stress and apoptosis. RESULTS AND DISCUSSION: We now demonstrate that GP uteri were enlarged by both estradiol (E2) and medroxyprogesterone acetate (MPA) (p < 0.001). Effects of MPA on uterine weight differed significantly depending on E2 levels (p < 0.001), where MPA opposed the E2 effect in combined treatments. Angiogenesis parameters were similarly impacted upon: MPA alone increased BV density (p = 0.036) and BV average area (p = 0.002). The presence of E2 significantly decreased these parameters. These changes were associated with highly elevated of the lipid peroxidation product, 8-isoprostane (8-isoP) content in E2+MPA-treated and by nuclear 8-OH-deoxyguanosine (8oxoG) staining compared to all other groups (p < 0.001). Abnormalities in the E2+MPA group were consistent with chromatin redistribution, nuclear pyknosis, karyolysis and increased apoptosis as observed by a marked increase in TUNEL labeling. CONCLUSIONS: LTPOC exposure alters endometrial vascular and tissue morphology consistent with oxidative stress and apoptosis in a complex interplay with endogenous estrogens. These findings are remarkably similar to in vivo change observed in the human uterus following LTPOC administration. Hence, the GP is an excellent model for the study of LTPOC effects on the uterus and will be extremely useful in determining the mechanistic pathways involved in this process which cannot be conducted on humans

Localized Epigenetic Changes Induced by DH Recombination Restricts Recombinase to DJH Junctions

Immunoglobulin heavy chain (Igh) genes are assembled by sequential rearrangements of diversity (DH) and variable (VH) gene segments. Three critical constraints govern VH recombination. These include timing (VH recombination follows DH recombination), precision (VHs recombine only to DJH junctions) and allele specificity (VH recombination is restricted to DJH recombined alleles). We provide a model for these universal features of VH recombination. Analyses of DJH recombined alleles revealed that DJH junctions were selectively epigenetically marked, became nuclease sensitive and bound RAG proteins, thereby permitting DH-associated recombination signal sequences to initiate the second step of Igh gene assembly. We propose that VH recombination is precise because these changes did not extend to germline DH gene segments located 5′ of the DJH junction

Group versus individual academic detailing to improve the use of antihypertensive medications in primary care: a cluster-randomized controlled trial

Purpose To compare group versus individual academic detailing to increase diuretic or β-blocker use in hypertension. Methods We conducted a cluster-randomized controlled trial in a large health maintenance organization. Subjects (N=9820) were patients with newly treated hypertension in the year preceding the intervention (N=3692), the 9 months following the intervention (N=3556), and the second year following intervention (N=2572). We randomly allocated 3 practice sites to group detailing (N=227 prescribers), 3 to individual detailing (N=235 prescribers), and 3 to usual care (N=319 prescribers). Individual detailing entailed a physician-educator meeting individually with clinicians to address barriers to prescribing guideline-recommended medications. The group detailing intervention incorporated the same social marketing principles in small groups of clinicians. Results In the first year following the intervention, the rates of diuretic or β-blocker use increased by 13.2% in the group detailing practices, 12.5% in the individual detailing practices, and 6.2% in the usual care practices. As compared with usual care practices, diuretic or β-blocker use was more likely in group detailing practices (adjusted odds ratio (OR), 1.40; 95% confidence interval (CI), 1.11 – 1.76) and individual detailing practices (adjusted OR, 1.30; 95% CI, 0.95 – 1.79). Neither intervention affected blood pressure control. Two years following this single-visit intervention, there was still a trend suggesting a persistent effect of individual (OR, 1.22; 95% CI, 0.92 – 1.62), but not group, detailing (OR, 1.06; 95% CI, 0.80 – 1.39), as compared with usual care. Conclusion Both group and individual academic detailing improved antihypertensive prescribing over and above usual care but may require reinforcement to sustain improvements

Multiethnic Societies of Central Asia and Siberia Represented in Indigenous Oral and Written Literature

Central Asia and Siberia are characterized by multiethnic societies formed by a patchwork of often small ethnic groups. At the same time large parts of them have been dominated by state languages, especially Russian and Chinese. On a local level the languages of the autochthonous people often play a role parallel to the central national language. The contributions of this conference proceeding follow up on topics such as: What was or is collected and how can it be used under changed conditions in the research landscape, how does it help local ethnic communities to understand and preserve their own culture and language? Do the spatially dispersed but often networked collections support research on the ground? What contribution do these collections make to the local languages and cultures against the backdrop of dwindling attention to endangered groups? These and other questions are discussed against the background of the important role libraries and private collections play for multiethnic societies in often remote regions that are difficult to reach

Multiethnic Societies of Central Asia and Siberia Represented in Indigenous Oral and Written Literature

Central Asia and Siberia are characterized by multiethnic societies formed by a patchwork of often small ethnic groups. At the same time large parts of them have been dominated by state languages, especially Russian and Chinese. On a local level the languages of the autochthonous people often play a role parallel to the central national language. The contributions of this conference proceeding follow up on topics such as: What was or is collected and how can it be used under changed conditions in the research landscape, how does it help local ethnic communities to understand and preserve their own culture and language? Do the spatially dispersed but often networked collections support research on the ground? What contribution do these collections make to the local languages and cultures against the backdrop of dwindling attention to endangered groups? These and other questions are discussed against the background of the important role libraries and private collections play for multiethnic societies in often remote regions that are difficult to reach

Targeting cap-dependent translation blocks converging survival signals by AKT and PIM kinases in lymphoma

PIM kinase expression in human lymphomas can influence the outcome of chemotherapy, and blocking cap-dependent translation can reverse PIM-mediated rapamycin resistance in murine lymphomas

Factors That Influence the Quality of RNA From the Pancreas of Organ Donors

Attaining high-quality RNA from the tissues or organs of deceased donors used for research can be challenging due to physiological and logistical considerations. In this investigation, METHODS: RNA Integrity Number (RIN) was determined in pancreatic samples from 236 organ donors and used to define high (≥6.5) and low (≤4.5) quality RNAs. Logistic regression was used to evaluate the potential effects of novel or established organ and donor factors on RIN. Univariate analysis revealed donor cause of death (odds ratio [OR], 0.35; 95% confidence interval [CI], 0.15-0.77; P = 0.01), prolonged tissue storage before RNA extraction (OR, 0.65; 95% CI, 0.52-0.79; P < 0.01), pancreas region sampled (multiple comparisons, P < 0.01), and sample type (OR, 0.32; 95% CI, 0.15-0.67; P < 0.01) negatively influenced outcome. Conversely, duration of final hospitalization (OR, 3.95; 95% CI, 1.59-10.37; P < 0.01) and sample collection protocol (OR, 8.48; 95% CI, 3.96-19.30; P < 0.01) positively impacted outcome. Islet RNA obtained via laser capture microdissection improved RIN when compared with total pancreatic RNA from the same donor (ΔRIN = 1.3; 95% CI, 0.6-2.0; P < 0.01). A multivariable model demonstrates that autopsy-free and biopsy-free human pancreata received, processed, and preserved at a single center, using optimized procedures, from organ donors dying of anoxia with normal lipase levels increase the odds of obtaining high-quality RNA

Toll-like Receptor 9, Maternal Cell-free DNA and Myometrial Cell Response to CpG Oligodeoxynucleotide Stimulation

Problem: Among mechanisms triggering onset of parturition, it has been recently postulated that Toll-Like Receptor (TLR)9 engagement by cell-free DNA (cfDNA) triggers inflammation, myometrial contractions, and labor in absence of infection. The current study evaluated whether direct (myometrial) or indirect (decidual) TLR9 engagement enhances human myometrial contractility. Method of Study: Toll-like receptor 9 expression and cellular localization were surveyed by immunohistochemistry of placenta, fetal membranes, and myometrium in term (gestational age [GA]: \u3e37 weeks) labor (TL, n = 7) or term non-labor (TNL, n = 7) tissues. Non-pregnant myometrium (n = 4) served as reference. TLR9 mRNA expression relative to other TLRs was evaluated through the mining of an RNA-seq dataset and confirmed by RT-PCR. Immortalized human myometrial cells (hTERT-HM) were treated with incremental concentrations of TLR9 agonist ODN2395, TNF-α, or LPS. Secreted cytokines were quantified by multiplex immunoassay, and contractility was assessed by an in-gel cell contraction assay (n = 9). Induction of hTERT-HM contractility was also evaluated indirectly following exposure to conditioned media from primary term decidual cells (n = 4) previously stimulated with ODN2395. Results: Toll-like receptor 9 immunostaining in placenta and amniochorion was strongest in decidual cells, but unrelated to labor. TLR9 staining intensity was significantly decreased in TL compared with TNL myometrium (P = 0.002). Although total cfDNA in maternal circulation increased in TL (P = 0.025 vs TNL), difference in cffDNA was non-significant. Myometrial TLR9 mRNA levels were unaffected by contractile status and far less abundant than other pro-inflammatory TLRs. hTERT-HM contractility was enhanced by LPS (P = 0.002) and TNF-α (P = 0.003), but not by ODN2395 (P = 0.345) or supernatant of TLR9-stimulated decidual cells. Conclusion: Myometrial and decidual TLR9 are unlikely to directly regulate human parturition