examination of how LPS may influence the follicular fluid surrounding the oocyte and to parallel a follicular phase HS trial performed by our group (Dickson et al., under review). We were successful in infusing a dose of LPS that was high enough to elicit an initial response in levels of insulin and LPS binding protein but not so high as to induce a fever. These differences existed on day 1 but not day 3 or 5 of the infusion, indicating a development of tolerance. Additionally, there was no difference in LBP levels in follicular fluid. Gilts in the LPS group had smaller follicular diameter than the control gilts. It is not known if this is a direct effect of LPS exposure on the follicle or an indirect result of the immune system using a greater partition of energy to deal with inflammation. Interestingly, we saw no changes in abundance of PI3K pathway components measured, even though there was an increase in ovarian TLR4 abundance. The possibility that temporal changes in these pathway components may have occurred earlier in the infusion timeline cannot be discounted. Additionally, acyloxyacyl hydrolase (AOAH) was found in ovaries of both groups. This lipase is capable of cleaving the lipid A moiety from LPS, rendering it unable to effectively bind TLR4 and elicit an inflammatory response . Though we saw no differences between the groups, the finding is novel and warrants further investigation into potential detoxification of LPS in the ovary.
Previous studies investigating effects of hypoglycemia on platelet biology have suggested an increase in platelet reactivity; however, this increase was in the context of hypoglycemic stimulus as part of an insulin stress test (41). An older investigation into the effect of hypogly- cemia on monocyte-platelet interactions in type 1 di- abetes and healthy controls also suggested a trend toward increased MPA formation, but these data were not conclusive, with little difference between euglycemic and hypoglycemic conditions (42). Our study also recapitulates and extends previous findings that hypoglycemia is pro- thrombotic, as evidenced by an increased platelet count and increased platelet reactivity to ADP (43). We have con- clusively demonstrated an overall increase in formation of MPAs in hypoglycemia in comparison with euglycemia. Furthermore, we provide data demonstrating MPA formation within monocyte subsets in experimental hypoglycemia. MPA formation is a highly sensitive marker of both monocyte and platelet activation (44, 45). MPA formation promotes monocyte release of the proinflammatory cytokines TNF a , C-X-C motif che- mokine ligand 8, and C-C motif chemokine ligand 2 (46, 47) and increases adhesive properties of monocytes
Many tumour cells are unable to survive as single cells under anchorage- independent growth conditions; thus they form aggregates to avoid anoikis [365, 374, 375]. EOC cells that are shed from the primary tumour, and forced into suspension within the abdominal cavity, are believed to aggregate as multicellular spheroids in order to maintain cell-cell contact as part of their natural survival response . Key cell-cell adhesion molecules such as cadherins and integrins facilitate cell compaction and create a tumour microenvironment that supports mechanisms of cell survival [363, 376, 377]. The formation of ovarian cancer spheroids is greatly disrupted, for example, when cells are treated with blocking antibodies against α5- or β1-integrin subunits . Furthermore, the formation of multicellular aggregates is likely an important intermediate mechanism that facilitates metastasis. It has been suggested that spheroids should be considered in the dissemination of EOC and that the adhesion of spheroids initiates the conversion of floating cells within the ascites to an anchored metastatic lesion . Of note, when spheroids implant on the mesothelium (a cell monolayer that covers all organs in the peritoneal cavity) they do not directly adhere to mesothelial cells, but rather to the ECM underneath [364-366]. The Brugge laboratory has demonstrated that ovarian cancer spheroids use myosin-generated force to displace the mesothelium, thereby gaining access to the ECM to promote invasion . Burleson and colleagues have also shown that ovarian cancer spheroids adhere to, migrate on and invade into live human
surrounded by granulosa cells (GCs), theca cells, a fluid-filled antrum, and a basal lamina separating GCs from TCs, grows and differentiates, culminating in development of a fertilizable oocyte. TCs under the influence of luteinizing hormone (LH) produce androgens. GCs under the influence of follicle stimulating (FSH) hormone proliferate, produce steroid hormones, and differentiate to become responsive to the surge of LH that initiates ovulation. The cellular processes of folliculogenesis require cell-extracellular matrix (ECM) interactions. Spondin 1 (SPON1) is an ECM protein primarily studied for its role in nervous system development as a nerve outgrowth signalling molecule, but is also known to affect cell viability, differentiation, and migration in non-neural tissues and cells. Evidence that Spondin 1 is functional within the ovary includes its discovery in bovine follicular fluid, its increased mRNA expression in response to estrogen in uterus and mammary gland, its decreased mRNA expression in GCs of mice null for estrogen receptor β, and its overexpression in ovarian cancer. Despite the possible importance of Spondin 1 in the ovary it has never been characterized in this tissue. This study was undertaken with the goal of elucidating roles of Spondin 1 in the ovary. Experiments with the human GC tumour cell line, KGN, found that Spondin 1 increases cell viability and proliferation possibly by activating the mTORC1 complex, and decreases cAMP-induced progesterone production by inhibiting cAMP-induced STAR transcription. Experiments with mouse primary GCs corroborated the effects of Spondin 1 on granulosa cell viability and again found a role for Spondin 1 in steroidogenesis, however, progesterone
miR-21* acts as a driver of proliferation on induction by cisplatin. The effect was phenocopied by NAV3 knockdown; NAV3 knockdown by itself did not induce any changes in proliferation; however on cisplatin treatment, there is a significant increase in proliferation. These indicate that miR-21* drives proliferation on cisplatin treatment. One of the limitations of the project is that the actual mechanism of action of miR-21* and NAV3 has not been identified. Another limitation is that most of the experiments have been carried out in vitro. Other considerations include the fact that the effect of knockdown of gene function by the siRNAs was not confirmed in all the cell lines; also the efficacy of the drug 16F16 on PDIA4 was not quantified. There is much scope for future research into the actual mechanism by which miR-21* and NAV3 modify response to cisplatin and if this is the same in all the cell lines and tumours. More of the genes identified as being of interest could be followed up; pathway analysis could also be used to identify the actual mechanism. It would be interesting to study if this effect on cisplatin resistance is also present to other drugs such as paclitaxel as well as in other types of cancers. Another avenue to explore would include studying the levels in blood of patients with ovarian cancer to identify if this could distinguish some patients who are resistant, thereby predicting chemotherapy response.
During inflammation, many cells, predominantly macrophages, produce cytokines such as IL-1β, IL-6, and TNF-α at inflammation sites. These cytokines stimulate production of plasma proteins, also known as acute phase proteins (APP), in the liver. These APP include LBP, serum amyloid A (SAA), and Haptoglobin (Hp). SAA is an acute phase protein which exports cholesterol from phagocytosed cell membranes to enhance the availability of free phospholipids and cholesterol for new cell membranes needed during acute inflammation (Manley et al., 2006). Amyloid A was discovered in 1971 (Benditt et al., 1971) and the serum component, SAA was discovered shortly thereafter (Husby and Natvig, 1974), and interestingly, the structure has been conserved for some 500 million years (Santiago et al., 2000). At the initiation of inflammation, SAA rapidly increases from 100-1,000-fold (Epstein et al., 1999). Hp, an APP, plays an integral role in binding hemoglobin iron to render it inaccessible to bacteria (Eaton et al., 1982). It is known that elevated serum APP is a response to metritis (Smith et al., 1998; Hirvonen et al., 1999) as well as retained placenta (Mordark, 2009). Cows with acute puerperal metritis display increased serum Hp and SAA compared to healthy cows and the cows with elevated Hp took longer to conceive in their subsequent pregnancy, indicating that subclinical uterine infection could contribute to poor conception rates and decreased reproductive efficiency (Ceciliani et al., 2012).
Our results are in accordance with previous in vitro and animal experiments on the immunomodulating effect of nucleoside transport inhibition. Dipyridamole enhances the LPS-induced IL-10 production  and attenuates the production of TNF-a  and other proinflammatory cytokines in human cultured mono- nuclear cells. Furthermore, dipyridamole therapy in patients undergoing coronary artery bypass grafting inhibited postoperative ex vivo polymorphonuclear cell adhesion to endothelial cells . Also in animal stu- dies, administration of ENT inhibitors limited the inflammatory response and reduced tissue injury in situations of severe inflammation [9-11]. Of importance, these effects were abolished by concomitant administra- tion of adenosine A 2a receptor antagonists .
"Data presented in this study dealt with 81 women enrolled in IVF programs. The mean age of the cases in the present study (28.7 years) was close to that reported in Egyptian (29 years) and Iranian (29.1 years) studies (13-16), but lower than that reported from the Netherlands (33.8 years)" (5, 16). The younger age of women seeking IVF in developing countries, including Gaza Strip, could be explained in the context of social habits where people marry at young age and the desire to have children immediately after marriage. Women undergoing IVF showed that ovarianresponse is better with decreasing age, showing poor response (<4 oocytes) at the oldest age (36.5 years) and high response (>16 oocytes) at the youngest age (26.3 years). Consequently younger women have a better chance of a successful IVF. This inverse relationship between women response and age was also documented in other studies (15-19).
Selection of relevant predictors and estimation of param- eters were performed on a dataset with a limited number of events (i.e., poor response: n ⫽ 36; also see the results section). Therefore, to exclude overoptimism when applying the final logistic model and to assess the stability of the selection of variables, a correction was performed (18). In this so-called bootstrap sampling procedure, the selection and estimation process was repeated 200 times after creating 200 new datasets of 120 cases (bootstrap samples) by draw- ing cases with replacement from the original data. The original statistical analysis was performed on each bootstrap sample, yielding 200 sets of predictors and parameter esti- mates. The model estimates of each bootstrap sample were evaluated on the original data and a new mean ROC AUC was calculated, now corrected for statistical overoptimism and thus yielding a more accurate estimate of future perfor- mance.
Women, aged 20 to 39 years with infertility and oligomenorrhea were recruited for this study from infertility clinic at Aziz Medical Centre in Kara- chi, Pakistan, from 1st August 2015 to 31st July 2016. Infertility was defined as the inability to conceive after 24 months of regular unprotected sexual intercourse where semen analysis was re- ported within normal limits. Women with oligo- menorrhea which was defined for the study pur- pose as at least three menstrual cycle lengths of more than 40 days during the preceding year were selected as target population. Excluded from the study were those with history of cancer treatment, known autoimmune disorders, prior treatment of endometriosis or surgery to reproductive tract, prior history of pelvic inflammatory disease, those with tubal factors and uterine factors as assessed on history and confirmed by normal hysterosal- pingogram. Also, women who satisfied the Rot- terdam’s criteria for polycystic ovarian syndrome (PCOS) were excluded from the study (11).
We investigated neurophysiological responses to un- pleasant gustatory stimuli based on S1-S2 contingent ne- gative variation (CNV) paradigm, to clarify the intrinsic food preference mechanism. The CNV was one of the first ERP components to be described and it is a slow ne- gative-going ERP elicited by a warning stimulus that re- quires anticipation of a target stimulus [7,8]. After disco- very of the CNV, Loveless and Sanford (1975) and Weerts and Lang (1973) suggested that the component is quanti- fiable into two distinct subcomponents: an “early” CNV and a “late” CNV [9,10]. The presence of the early CNV is generally thought to be a cortical reflection of control- led, rather than automatic, psychological processes in response to an S1 that requires anticipation of a subse- quent S2 [7,11], while the late CNV is measured just prior to the onset of the target stimulus, and reflects the additional contribution of cortical resources required for motor response preparation [12,13]. To date, there are many researches which describe what stimulus characte- ristics can affect the characteristics of the CNV. For ex- ample, attention and expectancy, intensity, modality, du- ration, stimulus rate, probability, stimulus relevance and pitch discrimination can affect the CNV component [14, 15]. However, there also exist some debates regarding which cognitive processing the CNV is associated with. Considerable evidence suggested that the early compo- nent of the CNV of the ERP, thought to reflect anticipat- ed cognitive effort, is sensitive to the effortful process that contributes to the manifestation of prejudice . Since several patterns of cognitive processes are measurable by the early, late CNV may be related to the expression of the prejudice. In the present study, we expected that the S1-S2 CNV paradigm would be adapted to quantify the cognitive processing during taste processing for investi- gating neural mechanisms of taste preference.
Chromatin undergoes structural changes in response to extracellular and environmental signals. We observed changes in nuclear morphology in cancer tissue biopsied after chemotherapy and hypothesised that these DNA damage-induced changes are mediated by histone deacetylases (HDACs). Nuclear morphological changes in cell lines (PE01 and PE04 models) and a xenograft model (OV1002) were measured in response to platinum chemotherapy by image analysis of nuclear texture. HDAC2 expression increased in PEO1 cells treated with cisplatin at 24h, which was accompanied by increased expression of heterochromatin protein 1 (HP1). HDAC2 and HP1 expression were also increased after carboplatin treatment in the OV1002 carboplatin-sensitive xenograft model but not in the insensitive HOX424 model. Expression of DNA damage response pathways (pBRCA1, γH2AX, pATM, pATR) showed time-dependent changes after cisplatin treatment. HDAC2 knockdown by siRNA reduced HP1 expression, induced DNA double strand breaks (DSB) measured by γH2AX, and interfered with the activation of DNA damage response induced by cisplatin. Furthermore, HDAC2 depletion affected γH2AX foci formation, cell cycle distribution, and apoptosis triggered by cisplatin, and was additive to the inhibitory effect of cisplatin in cell lines. By inhibiting expression of HDAC2, reversible alterations in chromatin patterns during cisplatin treatment were observed. These results demonstrate quantifiable alterations in nuclear morphology after chemotherapy, and implicate HDAC2 in higher order chromatin changes and cellular DNA damage responses in ovarian cancer cells in vitro and in vivo.
Statistical analysis was done using JMP Software (SAS Institute, SAS Campus Diveway, Cary, North Carolina 27513). The effect of progesterone on each response variable was done using a regression model for the continuous variables, follicle number, CL size, cleavage rate, and embryo rate. For each categorical variable (pregnancy, breed, follicular response, and cyst presence) the mean and standard deviation were found using a one-way ANOVA. For the effect of multiple variables on the response variable (ie progesterone, pregnancy, and CL size on total follicular number) a fit model was constructed and the parameter estimates for each variable were considered. Not all data was independent because cows were used multiple times. The effects of this were considered in a multivariate analysis and found not to be significantly different from the fit model.
In our study, participants gave their written informed consent after a detailed explanation regarding the mean- ing of ovarian deserve and information about the pos- sible beneficial effects and potential side effects of DHEA. All procedures and sample collection methods were approved by the Human Ethics Committees of Beijing Obstetrics and Gynecology Hospital, Capital Medical University. The subfertile women undergoing IVF or intracytoplasmic sperm injection-embryo transfer (ICSI-ET) therapy were due to tubal and/or male factors or failure of intrauterine insemination (IUI) for 3 times. The inclusion criteria included  age <40 years,  duration of subfertility >1 year, and  DOR, diagnosis criteria including the following two or more items such as 25 IU/L > FSH >10 IU/L, E 2 > 80 pg/ml, AMH
Severe endotoxemia, a condition where microembolization and intravascular coagulation are thought to play important roles, was treated experimentally with prostacyclin (PGI 2 ). In a study of 24 dogs, 8 control animals injected with 1.75 mg·kg −1 of endotoxin died within 24 h. Six animals given intravenous aspirin 100 mg/kg, 30 min after endotoxin died. 9 of 10 dogs infused with 100 ng PGI 2 ·kg −1 ·min −1 for 3 h, given 30 min after the injection of endotoxin survived 24 h (P < 0.025). Injection of endotoxin resulted in a: (a) maximal 62% fall in mean arterial pressure (P < 0.001); (b) transient doubling of mean pulmonary arterial pressure (P < 0.001); (c) initial 70% drop in cardiac index (P < 0.001); (d) decline in blood platelets from 213,700 to 13,700/mm 3 (P < 0.001), and leukocytes from 7,719 to < 750/mm 3 (P < 0.001); (e) depressed urine output (P < 0.001); (f) 34% decrease in blood fibrinogen (P < 0.01) and an increase in fibrin degradation products > 50 µg/ml (P < 0.001); (g) fivefold increase in circulating cathepsin D titer (P < 0.005) and (h) increase in blood norepinephrine (P < 0.005), dopamine (P < 0.005), and epinephrine (P < 0.001). Aspirin treatment led to an increase in mean arterial pressure (P < 0.001) and mean pulmonary arterial pressure (P < 0.005), but cardiac index, […]
changes at both time points were significant, the higher of the two values is shown). Positive direction shows an increase in concentration, while negative shows a decrease. Accordingly, the concentrations of metabolites in the first and third quadrants change in parallel with the observa- tions in endotoxemia; while the ones in the second and fourth quadrants change in the opposite direction. The response reflected by the direction and magnitude of change in septic patients agrees well with response to LPS within this common subset. However, for the SIRS group, the directions of change are not in agreement with those in endotoxemia for more than half of the metabolites. This suggests that, at the whole body metabolome level, SIRS elicits a unique response with distinctive features. One such feature is the marked decrease in sulfated androgenic hormones (epiandrosterone sulfate, androsterone sul- fate, dehydroisoandrosterone sulfate (DHEA-S), 5alpha- pregnan-3beta,20alpha-diol disulfate) (Additional file 3: Table S2). Lower plasma concentration of one of these metabolites, DHEA-S, has previously been associated with other systemic inflammatory diseases, such as systemic lupus erythematosus and inflammatory bowel disease . This supports the idea that the inflammatory response without apparent infectious stimuli might elicit distinctive features not shared with sepsis or endotoxemia. It has been previously suggested that acute inflammatory stresses from different etiologies result in highly similar responses in humans at the genomic level . The observed distinct metabolomic responses to systemic inflammation with or without confirmed infection, however, suggest that the metabolome is much better at differentiating and understanding the various path- ophysiologies of the different systemic inflammatory responses. Identified unique features of the inflammatory response in different contexts may aid in improving the diagnosis or the development of more targeted therapies.
Firstly, we tested their cytotoxicity in vitro using the SKOV3 ovarian cancer cell line, and found that both B01002 and C26001 could inhibit tumor cell proliferation effectively (IC50 = 7.65 and 11.68 µ g/mL, respectively). Moreover, we found that both compounds could induce apoptosis of tumor cell, which seems the main mechanism of the toxicity, as a very small percentage of necrotic cells was observed. To evaluate the translational potential of these new agents, we tested their in vivo efficacy and safety using a xeno- graft mouse model. Consistent with the in vitro results, our study showed that both compounds decreased the size of xenografted tumors. By Ki-67 staining and PCNA level, we showed that both compounds inhibited the tumor prolifera- tion in vivo. By measuring body weight, food consumption, observing clinical signs, physical examination, blood routine testing, and biochemical tests, as well as necropsy of major organs, the mice showed good tolerance in various dosages to both compounds, indicating their clinical translational potential in future applications.
Establishing measurement invariance is essential for drawing accurate and valid infer- ences concerning respondents’ trait levels and for conducting comparisons between respond- ents from different groups. This dissertation investigated three aspects of measurement invar- iance in detail, namely 1) individual differences in response styles, 2) the measurement invar- iance of items between subgroups of respondents, and 3) the measurement invariance of items across assessment periods. If measurement invariance is violated regarding these three factors, trait scores do not represent the construct of interest purely. Instead, the measurement of the construct is contaminated by one or several secondary dimensions that influence item re- sponses in addition to the respondent’s latent trait level. The findings of this dissertation em- phasize that individual differences in response styles play an important role in questionnaire data since they explain variance in item responses that is incremental to the variance explained by the trait. This implies that trait scores may be biased and in consequence trait inferences and comparisons between individuals and groups based on sum scores can lead to distorted conclusions. However, response styles appear to be largely consistent across scales and there is tentative evidence that they might also be stable over time. This raises the possibility of correcting trait scores for response style effects. Model-based approaches such as mixed Rasch models or multidimensional item response models are able to separate stylistic variance from substantive variance in the measurement of response styles and to separate variance due to response styles from trait variance. This allows them to provide trait estimates that are cor- rected for response style effects. Therefore, trait comparisons should only be conducted using trait estimates containing a model-based correction for response style effects.
For each subject, at least eight hours of raw EEG were available for visual analysis. All EEGs before LPS infusion were within the normal range. One hour after LPS infu- sion mild transient encephalopathic EEG changes in the theta range were present in one subject for 15 minutes, without associated cognitive impairment. Of note, this subject had a very low cytokine response during endotox- emia (TNF-α level of 169 pg/ml compared with the group mean of 814 ± 133 pg/ml, and IL-6 level of 508 pg/ml compared with the group mean of 1,111 ± 142 pg/ml) and an average cortisol response (0.29 to 0.67 μmol/l). The EEGs from the other 14 subjects remained within the normal range after LPS infusion, and no focal or epilepti- form abnormalities were found.
The commonest cause of PICU admission in our study was respiratory failure that was usually due to viral lower respiratory tract infection. The majority (75%) of children with a respiratory tract infection had endotoxemia on admission despite having low severity- of-illness scores and good outcome. Several studies have documented high levels of endotoxin in the lung and blood of patients with respiratory infection as well as those on artificial ventilation. Pulmonary-to-systemic translocation of LPS has been demonstrated in non-pro- tective ventilation strategy studies in experimental ani- mals , and an association between ventilator- induced lung injury, ventilator-associated pneumonia, and systemic LPS and cytokine levels has been reported . Although the lung has no endogenous microflora that could provide a source of LPS, it is likely that bac- terial colonization of the airway or lung may be a poten- tial source of LPS that can translocate systemically following mechanical ventilation. The high prevalence of endotoxemia in our patients seems to confirm this.