Sprague Dawley rats from different vendors vary in the modulation of prepulse inhibition of startle (PPI) by dopamine, acetylcholine, and glutamate drugs.

RATIONALE: Rodent vendors are often utilized interchangeably, assuming that the phenotype of a given strain remains standardized between colonies. Several studies, however, have found significant behavioral and physiological differences between Sprague Dawley (SD) rats from separate vendors. Prepulse inhibition of startle (PPI), a form of sensorimotor gating in which a low-intensity leading stimulus reduces the startle response to a subsequent stimulus, may also vary by vendor. Differences in PPI between rat strains are well known, but divergence between colonies within the SD strain lacks thorough examination. OBJECTIVES: We explored intrastrain variation in PPI by testing SD rats from two vendors: Envigo and Charles River (CR). METHODS: We selected drugs acting on four major neurotransmitter systems that have been repeatedly shown to modulate PPI: dopamine (apomorphine; 0.5, 1.5, 3.0 mg/kg), acetylcholine (scopolamine; 0.1, 0.5, 1.0 mg/kg), glutamate (dizocilpine; 0.5, 1.5, 2.5 mg/kg), and serotonin (2,5-Dimethoxy-4-iodoamphetamine, DOI; 0.25, 0.5, 1.0 mg/kg). We determined PPI and startle amplitude for each drug in male and female Envigo and CR SD rats. RESULTS: SD rats from Envigo showed dose-dependent decreases in PPI after apomorphine, scopolamine, or dizocilpine administration, without significant effects on startle amplitude. SD rats from CR were less sensitive to modulation of PPI and/or more sensitive to modulation of startle amplitude, across the three drugs. CONCLUSIONS: SD rats showed vendor differences in sensitivity to pharmacological modulation of PPI and startle. We encourage researchers to sample rats from separate vendors before experimentation to identify the most suited source of subjects for their specific endpoints.

Association of Left Vertebral Artery Hypoplasia with Posterior Circulation Stroke and the Functional Outcome of Patients with Atrial Fibrillation-Related Cardioembolic Stroke.

BACKGROUND AND PURPOSE: A cardiogenic embolus could reach the posterior circulation through the right vertebral artery because of a relatively larger diameter in cases of left vertebral artery hypoplasia. Hence, we investigated whether left vertebral artery hypoplasia is associated with cardiac embolisms with atrial fibrillation in the posterior circulation and its functional outcomes. MATERIALS AND METHODS: In this monocentric retrospective study, patients with acute cardioembolic stroke with atrial fibrillation were enrolled and underwent CT or neck MRA, which visualized the aortic arch and subclavian arteries. The laterality and size of vertebral artery hypoplasia were recorded. Posterior circulation stroke, basilar artery occlusion, and the functional outcomes after 3 months were investigated. RESULTS: This study included 407 patients; the patients with left vertebral artery hypoplasia experienced a higher rate of posterior circulation stroke (19 versus 73; 42.2% versus 20.2%; P = .001) and basilar artery occlusion (5 versus 10; 11.1% versus 2.8%; P = .005) than the patients without left vertebral artery hypoplasia. Multivariate analysis revealed that left vertebral artery hypoplasia showed an association with lower odds of achieving a good functional outcome 3 months after the stroke (OR = 0.4; 95% CI, 0.2-0.9; P = .027). CONCLUSIONS: Patients with cardioembolic stroke and left vertebral artery hypoplasia had posterior circulation stroke, basilar artery occlusion, and poor functional outcomes after 3 months.

Ultrasound-assisted desalination of crude oil: The influence of mixing extent, crude oil species, chemical demulsifier and operation variables.

Coalescence of water droplets in crude oil has been effectively promoted by chemical demulsifiers integrated with ultrasound. Temporary images of water droplets in W/O emulsions were directly monitored using a metallurgical microscope. Water droplets achieved expansion of 118% at 40 min ultrasonic irradiation time under well mixing conditions. However, water droplets in heavy crude oil undergo less aggregation than those in light crude oil, due to resistance of mobility in highly viscous fluid. In the absence of chemical demulsifiers, water droplets enveloped by native surfactants appeared to aggregate arduously because of occurrence of interfacial tension gradients. Influential significance analyses have been executed by a factorial design method on operation variables, including acoustic power intensity, operation temperature, ultrasonic irradiation time and chemical demulsifier dosages. In this work, the outcomes indicate that the optimal operating conditions for desalination of crude oil assisted by ultrasound were as follows: acoustic power intensity = 300 W, operation temperature = 90℃, ultrasonic irradiation time = 75 min and chemical demulsifier dosages = 54 mg/L. Besides, it was found that the most influential importance of operation parameter was temperature, followed with acoustic power intensity, ultrasonic irradiation time and chemical demulsifier dosages.

Cell-free culture supernatant of Lactobacillus curvatus Wikim38 inhibits RANKL-induced osteoclast differentiation and ameliorates bone loss in ovariectomized mice.

This study was conducted to investigate the inhibitory effects of the cell-free culture supernatant of Lactobacillus curvatus Wikim 38 (LC38-CS) on RANKL-induced osteoclast differentiation and bone loss in a mice model of ovariectomy-induced post-menopausal osteoporosis. LC38-CS inhibited the RANKL-induced differentiation of bone marrow-derived macrophages (BMDMs) into osteoclasts in a dose-dependent manner. F-actin ring formation and bone resorption were also reduced by LC38-CS treatment of RANKL-treated BMDMs. In addition, LC38-CS decreased the RANKL-induced activation of the TRAF6/NF-κB/MAPKs axis at the early stage and the expression of osteoclastogenesis-related genes in BMDMs treated with RANKL. PRMT1 and ADMA levels, new biomarkers for osteoclastogenesis, were decreased by LC38-CS treatment. The administration of LC38-CS increased bone volume and bone mineral density in ovariectomized mice in µ-CT analysis. These findings suggest that LC38-CS inhibited RANKL-induced osteoclast differentiation by the downregulation of molecular mechanisms and exerted anti-osteoporotic effects.

[Multidisciplinary classification of magnetic resonance imaging features of neuropsychiatric lupus].

OBJECTIVE: To investigate the clinical correlation between the manifestations of neuropsychiatric lupus (NPSLE) and brain magnetic resonance imaging (MRI). METHODS: Retrospective analysis of 65 neuropsychiatric lupus patients with brain MRI and clinical data from Peking University Third Hospital from January 2006 to October 2016, which was classified by rheumatologist, neurologists, and radiologists based on their brain MRI findings. The correlation between brain MRI findings and clinical manifestations was analyzed. RESULTS: The characteristics of the brain MRI of the 65 patients were divided into 6 categories: 16 cases (25%) with demyelination in the white matter, 15 cases (23%) with cerebrovascular disease, including 4 cases (6%) with large vascular disease and 11 cases (17%) with small vessel disease, 4 cases (6%) with inflammation, 4 cases (6%) with edema, 13 cases (20%) with multiple manifestation coexistence, and 13 cases (20%) without any abnormality. Except for 4 cases of brain MRI with edema, the clinical manifestations were only epileptic seizures, other patients had complex and diverse clinical manifestations, including epileptic seizures, lupus-like headaches, mental symptoms, blurred vision, peripheral neuropathy and disturbance of consciousness. The incidence of epileptic seizures in patients with edema of MRI is significantly higher than that of other patients, and the therapeutic response time is the shortest. CONCLUSION: Multidisciplinary collaboration divides the MRI findings of neuropsychiatric lupus patients into six categories. This classification method helps clinicians to predict and intervene early possible neuropsychiatric symptoms to guide clinical treatment.

Quantifying Rome symptoms for diagnosis of the irritable bowel syndrome.

BACKGROUND: Irritable bowel syndrome (IBS) is a common functional gastrointestinal disorder, diagnosed on symptom-based criteria. Many have reported discrepancies between formal Rome criteria and diagnoses made in clinical practice. The aim of the study was to explore whether a quantitative version of the Rome criteria would better represent a clinical diagnosis of IBS than the current dichotomous criteria for symptom measure. METHODS: As part of a large, case-control study, participants completed a validated bowel disease questionnaire. Rome criteria were analyzed based on 15 individual symptoms. Penalized logistic regression model with stepwise selection was used to identify significant symptoms of IBS which were independently associated with case-control status. KEY RESULTS: In cases with a clinical diagnosis of IBS, 347 (70%) met Rome criteria for IBS. Increasing number of Rome symptoms were found related to the odds of being diagnosed with IBS. Nearly half of the Rome-negative case group experienced infrequent symptoms suggesting milder disease. Five of 15 Rome symptoms were associated with predicting case-control status in the final model, with 96% correctly classified among Rome-positive cases, 76% for Rome-negative cases, and 91% for controls. CONCLUSIONS AND INFERENCES: Irritable bowel syndrome appears to be a spectrum disorder. Quantifying individual symptoms of Rome criteria has greater utility than the current application in representing the degree of IBS affectedness and appears to better reflect a clinical diagnosis of IBS applied by physicians. The use of a quantitative diagnostic Rome "score" may be helpful in clinical practice and research studies to better reflect the degree an individual is affected with IBS.

Clinical diagnosis of cerebral vasospasm after subarachnoid hemorrhage by using transcranial Doppler sonography.

OBJECTIVE: To investigate the diagnosis of cerebral vasospasm (CVS) after subarachnoid hemorrhage (SAH) by transcranial Doppler (TCD) sonography. PATIENTS AND METHODS: 90 patients with SAH admitted to the Department of Neurosurgery of Weifang People's Hospital from January 2016 to December 2016 were selected. TCD and digital subtraction angiography (DSA) were used to diagnose the prevalence of CVS in patients. The severity of disease was evaluated (improved-Fisher grading). Correlations between neurological status (Hunt-Hess grading) and the prevalence of CVS were analyzed. It turned out that the prevalence of CVS was 87.78% detected by DSA and was 83.33% detected by TCD, no significant difference was found between them (p > 0.05). RESULTS: The results of TCD showed that the gender, age, smoking, alcoholism, and history of hypertension had no significant correlations with the prevalence of CVS (p > 0.05). Blood flow velocity of patients was significantly higher at 4-6 days after the occurrence of SAH compared with the level at 1-3 days, reached the peak at 7-9 days, and decreased at 10-12 days after occurrence. Significant differences in the severity of the disease were found between patients with different improved-Fisher grades and different Hunt-Hess grades (p < 0.05). The prevalence of CVS was significantly increased after SAH (p < 0.05). CONCLUSIONS: TCD can dynamically detect the blood flow velocity of SAH patients, and can be used for the prediction and diagnosis of CVS after SAH.

miR-520h is crucial for DAPK2 regulation and breast cancer progression.

This corrects the article DOI: 10.1038/onc.2015.168.

Tumour cell-derived WNT5B modulates in vitro lymphangiogenesis via induction of partial endothelial-mesenchymal transition of lymphatic endothelial cells.

Metastasis of the cervical lymph nodes frequently leads to poor survival of patients with oral squamous cell carcinoma (OSCC). The underlying mechanisms of lymph node metastasis are unclear. Wingless-type MMTV integration site family, member 5B (WNT5B), one component of the WNT signal pathway, was markedly up-regulated in OSCC sublines with high potential of lymphatic metastasis compared to that in OSCC cells with low nodal metastasis. Increased WNT5B mRNA was demonstrated in human OSCC tissues in comparison with adjacent non-tumorous tissues. Interestingly, the high level of WNT5B protein in serum was associated with lymph node metastasis in OSCC patients. Knockdown of WNT5B expression in OSCC sublines did not affect tumour growth but impaired lymph node metastasis and tumour lymphangiogenesis of orthotopic transplantation. Conditioned medium from WNT5B knockdown cells reduced the tube formation of lymphatic endothelial cells (LECs). In contrast, recombinant WNT5B enhanced the tube formation, permeability and migration of LECs. In LECs stained with phalloidin, the morphology of those treated with recombinant WNT5B changed from flat to spindle-like. Recombinant WNT5B also increased α-smooth muscle actin and inhibited the expression of vascular endothelial-cadherin but retained characteristics of endothelial cells. The results suggest that WNT5B functions in the partial endothelial-mesenchymal transition (EndoMT). Furthermore, WNT5B-induced tube formation was impaired in the LECs following the knockdown of EndoMT-related transcription factor, SNAIL or SLUG. The WNT5B-induced expression of Snail or Slug was abolished by IWR-1-endo and Rac1 inhibitors, which are involved in the WNT/ß-catenin and planar cell polarity pathways, respectively. Collectively, the data suggest that WNT5B induces tube formation by regulating the expression of Snail and Slug proteins through activation of canonical and non-canonical WNT signalling pathways.

[Clinical analysis on effect of retroperitoneoscopic nephrectomy in elderly donors for renal transplantation].

OBJECTIVE: To evaluate the safety and effectiveness of retroperitoneoscopic donor nephrectomy in elderly donors for renal transplantation. METHODS: A retrospective analysis was conducted with 123 cases of retroperitoneoscopic living donor kidney transplantation in 309th Hospital of PLA from March 2011 to March 2014, including 44 elderly donors (age≥55 years) and 79 young to middle-aged donors (age <55 years). Comparisons were made in terms of postoperative complications in both donors and recipients, renal function recovery in the donors and function of graft in the recipients. RESULTS: The clinical baseline data of the two groups shows that glomerular filtration rate (GFR) of donors in the elderly donor group was lower than the young donor group (P=0.04). The 123 donors all underwent retroperitoneoscopic donor nephrectomy successfully. Postoperative complications in donors and recipients of both groups had no significant differences (P=0.60; P=1.00). In the elderly donor group, the mean serum creatinine level of donors was significantly higher than that in the young donors group [(115.8±22.3) vs (102.5±16.3) µmol/L, P<0.01] 3 days after operation; and estimated GFR (eGFR) was lower [(53.0±9.1)vs(59.6±8.3)ml·min(-1)·(1.73 m(2))(-1,) P<0.01]. Serum creatinine and eGFR of the two groups showed no significant differences one week and six months after surgery (all P>0.05). Four recipients in the elderly donor group had delayed graft function (DGF), 3 had acute rejection; 8 recipients in the young donor group had DGF, 5 had acute rejection; no statistically significant differences were observed between the 2 groups (both P=1.00). Recipients' eGFR were higher in the young donor group than in the elderly donor group at 1 week, 1 month, 3 months, 6 months and 12 months after surgery, but with no statistically significant differences(all P>0.05). After (27.8±12.6) months follow-up, 1 recipient in the elderly donor group died from pulmonary infection; two recipients in the young donor group had kidney dysfunction. Graft survival in the two groups showed no significant difference(P=0.95). CONCLUSIONS: Retroperitoneoscopic donor nephrectomy is safe and feasible for elderly donors. With careful preoperative evaluation, precise operation, and close postoperative monitoring and follow-up, it could provide satisfactory clinical outcome.

An activated factor VII variant with enhanced tissue factor-independent activity speeds wound healing in a mouse hemophilia B model.

UNLABELLED: Essentials Disorders of hemostasis can lead to delayed and defective wound healing. In hemophilia B (HB) mice, 7 days of Factor (F)IX or VIIa are needed to normalize wound healing. One dose of a highly active FVIIa variant (DVQ) restored normal wound closure time in HB mice. Coagulation factors with enhanced activity may acquire biological effects not due to hemostasis. SUMMARY: Introduction We have previously reported that hemophilia B (HB) mice have delayed healing of cutaneous wounds and alterations in wound histology. Administration of a single dose of either factor IX or recombinant activated FVII (rFVIIa) (NovoSeven) prior to wounding did not improve wound closure time or histology. The FVIIa analog DVQ (V158D, E296V and M298Q mutations) was designed to have higher tissue factor-independent activity than rVIIa. We hypothesized that a single dose of DVQ would be more effective in restoring wound healing in HB mice. Methods Cutaneous punch wounds were made on the backs of HB and wild-type mice, and the time to wound closure was monitored. HB mice were treated with a dose of rFVIIa (10 mg kg(-1) ) or DVQ (1 mg kg(-1) ) that corrected the tail bleeding time. Skin samples were taken at various time points after wounding, fixed, and stained, and the histology was examined. Results As previously reported, wound closure times in HB mice given one dose of rFVIIa were not improved over those in untreated HB mice. Surprisingly, healing times in HB mice treated with an equally hemostatic dose of DVQ were normalized to that in wild-type mice. However, DVQ did not correct all histologic abnormalities in HB mice. Conclusions As the doses of DVQ and rFVIIa were chosen to support comparable levels of hemostasis, our data suggest that the improved healing seen with DVQ is not solely attributable to its hemostatic activity. It is possible that the improved wound healing arises through the effect of DVQ on cell signaling mechanisms.

Effect of fabric mounting method and backing material on bloodstain patterns of drip stains on textiles.

Textiles may provide valuable bloodstain evidence to help piece together events or activities at violent crime scenes. However, in spite of over 75 years of research, there are still difficulties encountered in many cases in the interpretation and identification of bloodstains on textiles. In this study, we dripped porcine blood onto three types of fabric (plain woven, single jersey knit, and denim) that are supported in four different ways (hard, taut, loose, and semi-hard, i.e., fabric laid on denim). These four mounting methods represent different ways in which a textile may be present when blood from a violent act lands on it. This study investigates how the fabric mounting method and backing material affect the appearance of drip stains on textiles. We found that bloodstain patterns formed on fabric lying flat on a hard surface were very different from when the same fabric was suspended loosely. We also found that bloodstains formed on the technical back of single jersey knit were vastly different from those on the technical face. Interestingly, some drip stains showed blood passing through the textile and leaving a stain behind it that resembled insect stains. By observing, recording, and describing how a blood stained textile is found or presented at the scene, the analyst may be able to better understand bloodstains and bloodstain patterns on textiles, which could be useful to confirm or refute a witness's account of how blood came to be where it was found after a bloodshed event.

The Role of Wnt Signaling in Squamous Cell Carcinoma.

Aberrant Wnt signaling pathway is a common feature of tumors and also plays important roles in tumor progression and metastasis of many cancer types. Various lines of evidence suggest that genetic defects affect Wnt pathway components, as well as epigenetic mechanisms that modulate the suppressors of Wnt pathway in oral squamous cell carcinoma. Recently, the newly discovered microRNAs are important molecular regulators in gene expression through transcription and translation repression. They play fundamental roles in a wide spectrum of biological functions, including cancer. In this review, we aim to accumulate recent research findings on the roles of Wnt/ß-catenin signaling and discuss how microRNAs affect Wnt/ß-catenin signaling in oral squamous cell carcinoma tumorigenesis. Apparently, investigations into the role of microRNAs with regard to the Wnt pathway in oral squamous cell carcinoma may help in the development of better strategies for tumor treatment.

miR-520h is crucial for DAPK2 regulation and breast cancer progression.

MicroRNAs (miRNAs) are small RNAs that suppress gene expression by their interaction with 3'untranslated region of specific target mRNAs. Although the dysregulation of miRNAs has been identified in human cancer, only a few of these miRNAs have been functionally documented in breast cancer. Thus, defining the important miRNA and functional target involved in chemoresistance is an urgent need for human breast cancer treatment. In this study, we, for the first time, identified a key role of miRNA 520h (miR-520h) in drug resistance. Through protecting cells from paclitaxel-induced apoptosis, expression of miR-520h promoted the drug resistance of human breast cancer cells. Bioinformatics prediction, compensatory mutation and functional validation further confirmed the essential role of miR-520h-suppressed Death-associated protein kinase 2 (DAPK2) expression, as restoring DAPK2 abolished miR-520h-promoted drug resistance, and knockdown of DAPK2 mitigated cell death caused by the depletion of miR-520h. Furthermore, we observed that higher level of miR-520h is associated with poor prognosis and lymph node metastasis in human breast cancer patients. These results show that miR-520h is not only an independent prognostic factor, but is also a potential functional target for future applications in cancer therapeutics.

Changes in oral mucosal MUC1 expression and salivary hormones throughout the menstrual cycle.

OBJECTIVE: To investigate relationships among oral mucosal epithelial MUC1 expression, salivary stress markers, and female gonadal hormones throughout the menstrual cycle. SUBJECTS AND METHODS: Thirty healthy women (25.9 ± 2.1 years) with regular menstrual cycle were included. Unstimulated (UWS) and stimulated whole saliva (SWS) were collected during the menstrual cycle. The expression level of oral mucosal MUC1 was analyzed. 17ß-Estradiol, progesterone, dehydroepiandrosterone (DHEA), cortisol, chromogranin A (CgA), and blood contamination levels were measured from UWS and SWS. RESULTS: Significant positive correlations were observed between 17ß-estradiol and DHEA in UWS, cortisol and CgA in UWS, MUC1 expression and DHEA in SWS, and among cortisol, progesterone, and DHEA in UWS and SWS. Significant negative correlations were observed between MUC1 and cortisol/DHEA ratio in UWS and SWS. When each phase was analyzed individually, MUC1 expression showed significant negative correlations with cortisol, progesterone, and cortisol/DHEA ratio in UWS and with progesterone and cortisol/DHEA ratio in SWS during the mid-luteal phase. A significant negative correlation was also observed between MUC1 and cortisol/DHEA ratio in UWS during the late luteal phase. CONCLUSIONS: Stress-related psychoendocrinological interactions throughout the menstrual cycle resulted in a decrease in oral mucosal epithelial MUC1 expression and a weakening of oral mucosal defense.

Phase I study of nanoliposomal irinotecan (PEP02) in advanced solid tumor patients.

PURPOSE: To define the dose-limiting toxicity (DLT), maximum tolerated dose (MTD) and pharmacokinetics (PK) of PEP02, a novel liposome-encapsulated irinotecan, in patients with advanced refractory solid tumors. METHODS: Patients were enrolled in cohorts of one to three to receive escalating dose of PEP02 in a phase I trial. PEP02, from 60 to 180 mg/m(2), was given as a 90-min intravenous infusion, every 3 weeks. RESULTS: A total of 11 patients were enrolled into three dose levels: 60 (one patient), 120 (six patients) and 180 mg/m(2) (four patients). DLT was observed in three patients, one at 120 mg/m(2) (grade 3 catheter-related infection) and two at 180 mg/m(2) (grade 4 neutropenia lasting for >3 days in one, grade 4 hematological toxicities and grade 4 diarrhea in the other). MTD was determined as 120 mg/m(2). Comparing with those after free-form irinotecan in the literature, the dose-normalized PK of SN-38 (the active metabolite) after PEP02 was characterized by lower C max, prolonged terminal half-life and higher AUC but with significant inter-individual variation. One patient who died of treatment-related toxicity had significantly higher C max and AUC levels of SN-38 than those of the other three patients at 180 mg/m(2). Post hoc pharmacogenetic study showed that the patient had a combined heterozygosity genotype of UGT1A1*6/*28. Two patients had objective tumor response. CONCLUSIONS: PEP02 apparently modified the PK parameters of irinotecan and SN-38 by liposome encapsulation. The MTD of PEP02 monotherapy at 3-week interval is 120 mg/m(2), which will be the recommended dose for future studies.

A KRAS mutation status-stratified randomized phase II trial of gemcitabine and oxaliplatin alone or in combination with cetuximab in advanced biliary tract cancer.

BACKGROUND: Previous clinical trials have not proved that adding epidermal growth factor receptor inhibitors to chemotherapy confers a survival benefit for patients with advanced biliary tract cancer (ABTC). Whether the KRAS mutation status of tumor cells confounded the results of past studies is unknown. PATIENTS AND METHODS: ABTC patients stratified by KRAS status, Eastern Cooperative Oncology Group performance status, and primary tumor location were randomized 1 : 1 to receive GEMOX (800 mg/m(2) gemcitabine and 85 mg/m(2) oxaliplatin) or C-GEMOX (500 mg/m(2) cetuximab plus GEMOX) every 2 weeks. The primary end point was objective response rate (ORR). RESULTS: The study enrolled 122 patients between December 2010 and May 2012 (62 treated with C-GEMOX and 60 with GEMOX). Compared with GEMOX alone, C-GEMOX was associated with trend to better ORR (27% versus 15%; P = 0.12) and progression-free survival (PFS, 6.7 versus 4.1 months; P = 0.05), but not overall survival (OS, 10.6 versus 9.8 months; P = 0.91). KRAS mutations, which were detected in 36% of tumor samples, did not affect the trends of difference in ORR and PFS between C-GEMOX and GEMOX. The two treatment arms had similar adverse events, except that more patients had skin rashes, allergic reactions, and neutropenia in the C-GEMOX arm. Of patients with C-GEMOX, the presence of a grade 2 or 3 skin rash was associated with significantly better ORR, PFS, and OS. CONCLUSIONS: Addition of cetuximab did not significantly improve the ORR of GEMOX chemotherapy in ABTC, although a trend of PFS improvement was observed. The trend of improvement did not correlate with KRAS mutation status. CLINICAL TRIALS NUMBER: This study is registered at ClinicalTrials.gov (NCT01267344). All patients gave written informed consent.

WWOX dysfunction induces sequential aggregation of TRAPPC6AΔ, TIAF1, tau and amyloid ß, and causes apoptosis.

Aggregated vesicle-trafficking protein isoform TRAPPC6AΔ (TPC6AΔ) has a critical role in causing caspase activation, tau aggregation and Aß generation in the brains of nondemented middle-aged humans, patients with Alzheimer's disease (AD) and 3-week-old Wwox gene knockout mice. WWOX blocks neurodegeneration via interactions with tau and tau-phosphorylating enzymes. WWOX deficiency leads to epilepsy, mental retardation and early death. Here, we demonstrated that TGF-ß1 induces shuttling of endogenous wild-type TPC6A and TPC6AΔ in between nucleoli and mitochondria (~40-60 min per round trip), and WWOX reduces the shuttling time by 50%. TGF-ß1 initially maximizes the binding of TPC6AΔ to the C-terminal tail of WWOX, followed by dissociation. TPC6AΔ then undergoes aggregation, together with TIAF1 (TGF-ß1-induced antiapoptotic factor), in the mitochondria to induce apoptosis. An additional rescue scenario is that TGF-ß1 induces Tyr33 phosphorylation and unfolding of WWOX and its the N-terminal WW domain slowly binds TPC6AΔ to block aggregation and apoptosis. Similarly, loss of WWOX induces TPC6AΔ polymerization first, then aggregation of TIAF1, amyloid ß and tau, and subsequent cell death, suggesting that a cascade of protein aggregation leads to neurodegeneration.

YM155 down-regulates survivin and XIAP, modulates autophagy and induces autophagy-dependent DNA damage in breast cancer cells.

BACKGROUND AND PURPOSE: The aim of this study was to determine the potency and molecular mechanism of action of YM155, a first-in-class survivin inhibitor that is currently under phase I/II clinical investigations, in various drug-resistant breast cancers including the oestrogen receptor positive (ER(+) ) tamoxifen-resistant breast cancer and the caspase-3-deficient breast cancer. EXPERIMENTAL APPROACH: The potency of YM155 in SK-BR-3, MDA-MB-231, MCF7 and its tamoxifen-resistant sublines, TamR6, TamR7, TamR8, TamC3 and TamC6, were determined by MTT assay. Western blot analysis, flow cytometric analysis, reverse transcription-PCR, fluorescent microscopy and comet assay were used to determine the molecular mechanism of action of YM155 in different breast cancer cell lines. KEY RESULTS: YM155 was equally potent towards the parental ER(+) /caspase-3-deficient MCF7 breast cancer cells and its tamoxifen-resistant sublines in vitro. The ER(-) /HER2(+) SK-BR-3 breast cancer cells and the triple-negative/caspase-3-expressing metastatic aggressive MDA-MB-231 breast cancer cells were also sensitive to YM155 with IC50 values in the low nanomolar range. Targeting survivin by YM155 modulated autophagy, induced autophagy-dependent caspase-7 activation and autophagy-dependent DNA damage in breast cancer cells. Interestingly, YM155 also induced XIAP degradation and the degradation of XIAP might play an important role in YM155-induced autophagy in breast cancer cells. CONCLUSIONS AND IMPLICATIONS: YM155 is a potent survivin inhibitor that has potential for the management of various breast cancer subtypes regardless of the expression of ER, HER2 and caspase-3. Importantly, this study provides new insights into YM155's molecular mechanism of action and therapeutic potential in the treatment of tamoxifen-resistant breast cancer.