Assay for estrogen and progesterone receptors of breast cancer cell lines in monolayer culture.
A whole-cell assay for measuring estrogen (ER) and progesterone (PgR) receptors in monolayer culture of human breast cancer cell lines is described. It is based on the measurement of incorporated tritiated ligands during 50 min of incubation (i.e. [3H]estradiol for ER, [3H]ORG-2058 for PgR). The assay fulfills all criteria of specificity as shown by competitive studies and measurements of the dissociation constants of the binding reactions. Moreover, a subcellular fractionation of MCF-7 labeled cells revealed that the majority of incorporated steroids was associated with the nuclear fraction. This finding is consistent with the concept of nuclear location of steroid-receptor complexes. Cultures in the presence of 10(-8) M estradiol indicated that the methodology is adequate for detecting the well-known estrogenic induction of PgR synthesis. The assay proved suitable for the quantitative assessment of the receptor content of various neoplastic (MCF-7; ZR-75-1, Cama-1, Evsa-T) and non-neoplastic (HBL-100) cell lines. The methodology has the other advantages of being simple and rapid, of requiring small amounts of cells and of allowing histological examination of the latter before, during and after biochemical analysis.


Lethal Effect of Total Dissolved Gas-Supersaturated Water with Suspended Sediment on River Sturgeon (Acipenser dabryanus).
High total dissolved gas (TDG) levels and excessive suspended sediment (SS) concentrations pose serious threats to fish survival during flood season. However, little information is available on the effects of TDG supersaturation with varying SS concentrations on fish. In this study, laboratory experiments were performed to investigate the effects of TDG supersaturation with varying SS concentrations on five-month-old river sturgeons (Acipenser dabryanus). The test fish were exposed to combinations of SS concentrations (0, 200, 600 and 1,000 mg/L) and TDG levels (125, 130, 135 and 140%), and their mortality and median lethal time (LT50) were quantified. The fish showed abnormal behaviors (e.g., quick breathing, fast swimming and an agitated escape response) and symptoms of gas bubble disease (GBD). SS increased the mortality of river sturgeon exposed to TDG supersaturation. Furthermore, the LT50 values at 125% TDG were 4.47, 3.11, 3.07 and 2.68 h for the different SS concentrations (0, 200, 600 and 1,000 mg/L, respectively), representing a significant decrease in LT50 with increasing SS. However, at higher TDG levels (130-140%), there was no significant increase in LT50 with increasing SS. Therefore, river sturgeon showed weak tolerance of TDG-supersaturated water with SS.


Central nervous system (CNS) enterovirus infections: A single center retrospective study on clinical features, diagnostic studies, and outcome.
Enteroviruses (EV) are responsible for a large number of meningoencephalitis cases, especially in children. The objective of this study was to identify modes of diagnosis including the significance of respiratory and cerebrospinal fluid samples, associated clinical characteristics, inpatient management, and outcome of individuals with EV infections of the central nervous system (CNS). Electronic medical records of individuals with enterovirus infections of the CNS who presented to the Columbia University Irving Medical Center and Children's Hospital of New York between January 1, 2012 and December 31, 2017 were reviewed retrospectively for demographic, epidemiological, and clinical data. The median age overall was 1.7 months (interquartile range 14 years) and most (62.4%) were male. The majority of CNS infections presented as meningitis (95.7%) and occurred in the summer (45.2%) and fall seasons (37.6%). Eighty-five cases (91.4%) demonstrated EV positivity in cerebrospinal fluid, thirty cases (32.3%) exhibited both cerebrospinal fluid and respiratory positivity, and eight cases (8.6%) exhibited respiratory positivity with coinciding neurological findings. Eighty-nine individuals overall (95.7%) received antibiotics and 37 (39.8%) received antiviral treatment. All surviving individuals had favorable Modified Rankin Scores (MRS) within the zero to two ranges upon discharge. Testing respiratory samples in addition to cerebrospinal fluid was found to be an important diagnostic tool in EV-associated cases. While clinical outcomes were favorable for an overwhelming majority of cases, etiological understanding of CNS infections is essential for identifying ongoing and changing epidemiological patterns and aid in improving the diagnosis and treatment.


Impact of uterine balloon tamponade on the use of invasive procedures in severe postpartum hemorrhage.
The aim of this study was to assess the impact of tamponade when uterotonic agents fail, on the need for surgery or interventional radiology. All women who received sulprostone for postpartum hemorrhage were retrospectively compared over two periods [December 2008 to December 2010 without use of tamponade (period 1) and June 2011 to June 2013 with use of tamponade (period 2)] in the case of sulprostone failure (STROBE compliant retrospective cohort study). During period 2, interventional radiology or surgery was used only in the case of tamponade failure. 165 women were included (74 for period 1, 91 for period 2). The rate of interventional radiology or surgery significantly decreased from period 1 (21 of 74 women, 28.4%) to period 2 (six of 91 women, 6.6%, p = 0.0003). The rate of assumed failure of uterotonic agents was higher for period 2: 22 of 74 women (29.7%) during period 1, and 41 of 91 (45.1%, p = 0.0439) during period 2. The success rate of tamponade was 92.1% (35 of 38 women). Although the efficacy of tamponade should be viewed in the light of its widespread use, our findings confirm that tamponade significantly reduces the need for interventional radiology or surgery for postpartum hemorrhage treatment.


Usefulness of low-dose dobutamine echocardiography to predict response and outcome in patients undergoing cardiac resynchronization therapy.
A substantial proportion of patients who meet the current guidelines for cardiac resynchronization therapy (CRT) fail to respond to this pacing modality. Although appropriate patient selection and left ventricular (LV) lead location have been ascribed as determinants of CRT response, the interaction among contractile reserve, dynamics of dyssynchrony, and lead location is not well understood. The present study prospectively evaluated the effect of contractile reserve and dobutamine-induced changes in LV synchrony, in relation to the LV lead location, as predictors of the response to CRT. In the present study, 31 patients were prospectively evaluated and underwent low-dose dobutamine echocardiography. The dobutamine-induced increase in ejection fraction (contractile reserve [CR]) was measured, and the most mechanically delayed segment was identified to classify patients into 2 groups. Group 1 had a CR of >20% and a LV lead position concordant with the mechanically delayed segment. Group 2 included the remaining patients (i.e., low CR, discordant LV lead position, or both). Patients in group 1 were significantly more likely to have an echocardiographic response at 6 months (80% of group 1 vs 29% of group 2, p = 0.018) and had an improved 2-year heart failure hospitalization-free survival rate (90% in group 1 vs 33% in group 2, p = 0.006). In conclusion, low-dose dobutamine echocardiography provides information that can help to predict responders to CRT. The response rates and heart failure hospitalization-free survival were improved in those patients with a CR >20% and an LV lead tip concordant with the most delayed mechanical segment.


Uptake of [3H]2-deoxy-D-glucose and [3H]dopamine in adrenal chromaffin cells of the mouse.
Deoxyglucose method was applied to adrenal chromaffin cells. It was shown that dichromate-fixation could fix [3H]deoxyglucose in tissue sections. Dichromate-fixation has two advantages as compared with glutaraldehyde-fixation: (1) two types of adrenal chromaffin cells, adrenaline-storing (A) cell and noradrenaline-storing (NA) cells, are distinguishable, (2) more [3H]deoxyglucose is retained in sections than in those fixed with glutaraldehyde. No difference was demonstrated in the uptake of deoxyglucose between A and NA cells while A cells incorporated more [3H]dopamine than NA cells. This indicates that such difference was not due to the different rate of glucose utilization.


Biodegradable pH-responsive micelles loaded with 8-hydroxyquinoline glycoconjugates for Warburg effect based tumor targeting.
Biodegradable triblock copolymer poly(ethylene glycol)-b-polycarbonate-b-oligo([R]-3-hydroxybutyrate) was prepared via metal-free ring-opening polymerization of ketal protected six-membered cyclic carbonate followed by esterification with bacterial oligo([R]-3-hydroxybutyrate) (oPHB). Amphiphilic triblock copolymer self-organizes into micelles with a diameter of ~25 nm. Acid-triggered hydrolysis of ketal groups to two hydroxyl groups causes an increase in hydrophilicity of the hydrophobic micelle core, resulting in the micelles swell and drug release. oPHB was added as core-forming block to increase the stability of prepared micelles in all pH (7.4, 6.4, 5.5) studied. Doxorubicin and 8-hydroxyquinoline glucose- and galactose conjugates were loaded in the micelles. In vitro drug release profiles in PBS buffers with different pH showed that a small amount of loaded drug was released in PBS at pH 7.4, while the drug was released much faster at pH 5.5. MTT assay showed that the blank micelles were non-toxic to different cell lines, while glycoconjugates-loaded micelles, showed significantly increased ability to inhibit the proliferation of MCF-7 and HCT-116 cells compared to free glycoconjugates. The glycoconjugation of anti-cancer drugs and pH-responsive nanocarriers have separately shown great potential to increase the tumor-targeted drug delivery efficiency. The combination of drug glycoconjugation and the use of pH-responsive nanocarrier opens up new possibilities to develop novel strategies for efficient tumor therapy.


Observation and rationalization of nitrogen oxidation enabled only by coupled plasma and catalyst.
Heterogeneous catalysts coupled with non-thermal plasmas (NTP) are known to achieve reaction yields that exceed the contributions of the individual components. Rationalization of the enhancing potential of catalysts, however, remains challenging because the background contributions from NTP or catalysts are often non-negligible. Here, we first demonstrate platinum (Pt)-catalyzed nitrogen (N2) oxidation in a radio frequency plasma afterglow at conditions at which neither catalyst nor plasma alone produces significant concentrations of nitric oxide (NO). We then develop reactor models based on reduced NTP- and surface-microkinetic mechanisms to identify the features of each that lead to the synergy between NTP and Pt. At experimental conditions, NTP and thermal catalytic NO production are suppressed by radical reactions and high N2 dissociation barrier, respectively. Pt catalyzes NTP-generated radicals and vibrationally excited molecules to produce NO. The model construction further illustrates that the optimization of productivity and energy efficiency involves tuning of plasma species, catalysts properties, and the reactor configurations to couple plasma and catalysts. These results provide unambiguous evidence of synergism between plasma and catalyst, the origins of that synergy for N2 oxidation, and a modeling approach to guide material selection and system optimization.


[Psychoanalytic considerations of right extremist orientation of male adolescents].
Giving a case example of an male adolescent with radical right-wing attitudes, the lack of male identification as well as in unresolved dyadic fixation to the mother will be shown. Though the radical right-wing orientation the adolescent tries on the basis of a phallic narcissistic conflict to establish bordes against an unresolved dyadic fixation to his mother.


Leveling the curve of Spee using different sized archwires: a randomized clinical trial of blood flow changes.
To compare blood flow (BF) changes of teeth subjected to orthodontic forces during curve of Spee (COS) leveling using different archwires (AW). Thirty subjects with COS > 5 mm were randomly assigned (1:1:1) into three groups based on the AW used: group 1: 0.017 × 0.025-inch stainless-steel (SS)AW, group 2: 0.019 × 0.025-inch SSAW, and group 3: 0.021 × 0.025-inch β-titanium (TMA)AW. In the 3 groups, a 5 mm-depth reverse COS was placed in the AWs. A laser Doppler flowmeter was used to measure BF at different time intervals (T0-T4). In the 3 AWs group, BF of all measured teeth was reduced 20 min after force application. Afterwards, the BF values started to increase until the baseline values were almost restored within 1 week. Differences in BF changes between the extrusion and intrusion subgroups were observed within groups 1 and 3 during the first 20 min of force application (P < 0.05). Similar BF changes were recorded using the 3 different AWs. BF changes were associated with tooth type and the amount of COS depth change. During CoS leveling, similar BF changes were recorded using the 3 different AWs. Tooth type and the amount of COS depth change were associated with BF changes within the first 20 min of force application. Greater BF reduction was found in premolars compared to incisors during the first 20 min of AW placement. It is important to select a type of applied forces that minimally affect the BF. Intrusive forces appeared to have lower negative effects on the BF of teeth during COS leveling. ClinicalTrial.gov (# NCT04549948).


Heparin-like anticoagulant polypeptides with tunable activity: Synthesis, characterization, anticoagulative properties and clot solubilities in vitro.
Due to the uncontrollable anticoagulant activity and limited source, Heparin, which is commonly used in clinical anticoagulation therapies, faces the risk of spontaneous bleeding and thrombocytopenia. Herein, a series of anionic poly(amino acid) s poly (l-Serine-ran-L-Glutamic acid-ran-L-Cysteine-SO3) (PSEC-SO3) were prepared by the controlled Ring Opening Polymerization (ROP) of N-Carboxyanhydrides (NCAs). The anticoagulant activities of PSEC-SO3 can be regulated by simply adjusting the feeding ratio of monomers. In vitro tests show that these polypeptides can effectively prolong the Activated Partical Thromboplastin Time (APTT) and inhibit Factor IIa and Factor Xa, but has no significant effect on Prothrombin Time (PT) and Thrombin Time (TT), which indicates that PSEC-SO3 mainly act on the intrinsic pathway. In summary, the activity-tunable heparin-like polypeptides are expected to have good application prospects in the anticoagulant field.


Creating highly amplified enzyme-linked immunosorbent assay signals from genetically engineered bacteriophage.
For early detection of many diseases, it is critical to be able to diagnose small amounts of biomarkers in blood or serum. One of the most widely used sensing assays is the enzyme-linked immunosorbent assay (ELISA), which typically uses detection monoclonal antibodies conjugated to enzymes to produce colorimetric signals. To increase the overall sensitivities of these sensors, we demonstrate the use of a dually modified version of filamentous bacteriophage Fd that produces significantly higher colorimetric signals in ELISAs than what can be achieved using antibodies alone. Because only a few proteins at the tip of the micron-long bacteriophage are involved in antigen binding, the approximately 4000 other coat proteins can be augmented-by either chemical functionalization or genetic engineering-with hundreds to thousands of functional groups. In this article, we demonstrate the use of bacteriophage that bear a large genomic fusion that allows them to bind specific antibodies on coat protein 3 (p3) and multiple biotin groups on coat protein 8 (p8) to bind to avidin-conjugated enzymes. In direct ELISAs, the anti-rTNFα (recombinant human tumor necrosis factor alpha)-conjugated bacteriophage show approximately 3- to 4-fold gains in signal over that of anti-rTNFα, demonstrating their use as a platform for highly sensitive protein detection.


Interleukin-6 improves the survival of mesencephalic catecholaminergic and septal cholinergic neurons from postnatal, two-week-old rats in cultures.
Interleukin-6 (human recombinant) supported the survival of cultured mesencephalic, catecholaminergic and septal cholinergic neurons from postnatal, two-week-old (P13-P15) rats. Significantly, more catecholaminergic neurons, stained by monoclonal anti-tyrosine hydroxylase antibody, were found in cultures supplemented with interleukin-6 at a concentration of 5 ng/ml than in cultures not treated with interleukin-6. The optimal dose used was 50 ng/ml. The survival effect of interleukin-6 on postnatal rat, tyrosine hydroxylase-positive neurons was observed both in cultures using serum-containing and serum-free medium. Contents of dopamine and noradrenaline in cultures with interleukin-6 were also larger than in control cultures. Interleukin-6 also increased the survival of cultured embryonic (E17) rat midbrain tyrosine hydroxylase-positive neurons. The effect on these neurons was, however, smaller, and the optimal dose of interleukin-6 was nearly 5 ng/ml. Interleukin-6 also supported the survival of cultured postnatal (P13) rat septal cholinergic neurons, visualized by acetylcholinesterase staining. The concomitant addition of mouse nerve growth factor (100 ng/ml) and interleukin-6 (50 ng/ml) had a synergetic effect on the survival of acetylcholinesterase-positive neurons in culture. Our data suggest that the survival of cultured tyrosine hydroxylase-positive, mesencephalic, and acetylcholinesterase-positive, septal neurons from postnatal two-week-old rats was supported by interleukin-6, just as there was a different dose dependency of interleukin-6 on the cultured postnatal neurons compared with embryonic neurons.


Ileal strangulation by a secondary perineal hernia after laparoscopic abdominoperineal rectal resection: A case report.
We report a recent case of strangulated bowel obstruction due to an incarcerated secondary perineal hernia that developed after laparoscopic rectal resection. A 75-year-old man undergoing treatment for alcoholic cirrhosis underwent laparoscopic abdominoperineal resection of the rectum (APR) for lower rectal cancer after preoperative chemoradiotherapy. Lung metastases were diagnosed 2 months postoperatively. Ten days after chemotherapy initiation, the patient was hospitalized on an emergency basis due to hepatic encephalopathy. Ten days thereafter, we observed perineal skin protrusion. Moreover, the skin disintegrated spontaneously, resulting in ascetic fluid outflow. Pain and fever developed, with inflammatory reactions. Contrast-enhanced computed tomography showed strangulated small bowel obstruction due to perineal hernia. We performed an emergency surgery, during which we found small intestine wall incarcerated in the pelvic dead space, with thickening and edema; no necrosis or perforation was observed. We performed internal fixation by introducing an ileus tube into the ileocecum and fixing its balloon at the cecal terminus. Secondary perineal hernia is rare and can develop after APR. Its prevalence is likely to increase in future because of the increasing ubiquity of laparoscopic APR, in which no repair of peritoneal stretching to the pelvic floor is performed. However, only two case of secondary perineal hernia causing strangulated bowel obstruction has been reported in the literature. The follow-up evaluation of our procedures and future accumulation of cases will be important in raising awareness of this clinical entity. We suggest that the pelvic floor and the peritoneum should be repaired.


Can vegetation provide shelter to cyprinid species under hydropeaking?
A novel study that focuses on the capacity of vegetation to provide shelter for fish species under hydropeaking regimes is presented. Two artificial patches mimicking the structure and density of Carex sp. mats were installed in an experimental flume to test whether submerged plants can offer flow refuge to two cyprinid species, Luciobarbus bocagei and Pseudochondrostoma polylepis, under baseflow and hydropeaking scenarios. Local flow fields were characterized using a Lateral Line Probe (LLP) and an Acoustic Doppler Velocimeter (ADV) and 33 h of video analysis were viewed to report, on a per-second basis, fish use of 1) patches with plants, 2) patches upstream and 3) downstream from vegetation and 4) patches without vegetation. Data on flow fields and fish patch use were integrated and analyzed to assess whether plants favored fish sheltering behavior. Vegetation created hydraulically stable areas suitable for fish to shelter, triggering changes in fish patch use. Although both species sheltered under hydropeaking, L. bocagei presented a stronger preference than P. polylepis for vegetated patches and areas downstream from plants, taking advantage of sheltered regions more frequently. P. polylepis weaker search for shelter could be related to species-specific factors and territorial behavior interferences rather than to fish performance relative to flume hydraulic conditions. Despite a weaker response, some P. polylepis individuals used patches downstream from plants more during the second half of the hydropeaking trials. A trade-off between reducing swimming effort and territoriality might explain this response. Results indicate that vegetation can help to counterbalance the impact of hydropeaking on fish while providing river functioning benefits. Evaluating fish sheltering to a wide set of river plants and patch designs on a species-by-species basis would help targeting vegetation-based actions for restoring hydropeaking rivers.


Which factors predict the loss of cervical lordosis following cervical laminoplasty? A review of various indices and their clinical implications.
Many patients undergoing laminoplasty develop postoperative loss of cervical lordosis or kyphotic alignment of cervical spine despite sufficient preoperative lordosis. This results in poor surgical outcomes. Here, we reviewed the relationship between multiple radiological parameters of cervical alignment that correlated with postoperative loss of cervical lordosis in patients undergoing laminoplasty. Patient with a high T1 slope (T1S) has more lordotic alignment of the cervical spine preoperatively and is at increased risk for the loss of cervical lordosis postlaminoplasty. Those with lower values of difference between T1S and Cobb's angle (T1S-CL) and CL-T1S ratio have higher risks of developing a loss of the cervical lordosis postoperatively. Alternatively, C2-C7 lordosis, neck tilt, cervical range of motion, and thoracic kyphosis had no role in predicting the postlaminoplasty kyphosis. Among various radiological parameters, the preoperative T1S is the most important factor in predicting the postoperative loss of the cervical lordosis/alignment following laminoplasty.


Chlorodibromomethane metabolism to bromide and carbon monoxide in rats.
The chlorodibromomethane (CDBM) metabolites bromide and CO were analysed as bromide level in plasma and carboxyhaemoglobin (COHb) level in blood of rats, respectively. The mean basic levels of bromide in plasma of rats receiving vehicle were 0.075 +/- 0.036 mmol/l (n = 27). After administration of CDBM at 0.4, 0.8, 1.6, and 3.1 mmol/kg p.o., the mean bromide levels rose to maximal values that were higher by a factor 27, 48, 69, and 135, respectively. Bromide elimination was slow and the plasma level was significantly increased following repeated administration in comparison to a single administration of CDBM. The CDBM concentrations in blood and in fat tissue 6 h after the last of 7 administrations of 0.8 mmol CDBM/ kg p.o., once a day for 7 consecutive days, were significantly lower than 6 h after a single gavage of this CDBM dose. The mean normal level of 0.45 +/- 0.32% COHb in rats (n = 30) was significantly increased following oral CDBM uptake. Initially higher COHb levels were measured after 7 consecutive applications of 0.8 mmol/kg CDBM. After a single administration of CDBM the level of glutathione disulphide in the liver was significantly increased; this effect was reversible. The oxidative CDBM metabolism was influenced by the glutathione (GSH) concentration in the liver. The rate of COHb and bromide formation was decreased after GSH depletion due to pretreatment of rats with buthionine sulphoximine (BSO) and increased following enhancement of the GSH concentration due to pretreatment of the animals with butylated hydroxyanisole (BHA). CDBM is a substrate for cytochrome P-450 2E1 (CYP2E1), as demonstrated by the inhibition of bromide and COHb formation due to simultaneous administration of CDBM and the CYP2E1 inhibitor diethyldithiocarbamate (DDTC); also by the initially higher levels of bromide in plasma and COHb in blood after gavage of CDBM pretreated with isoniazid (INH), an inducer of CYP2E1. The increase of bromide formation after CDBM administration in phenobarbital (PB)-pretreated rats indicated that cytochrome P-450 2B1 and 2B2 (CYP2B1 and CYP2B2) play a role as catalysts of the CDBM biotransformation. It is shown that m-xylene pretreatment, which activates CYP2E1 as well as CYP2Bs, leads to a higher bromide level after CDBM administration than the INH or PB pretreatment. In liver microsomes of rats treated with CDBM (0.8 mmol/kg p.o., seven daily applications), the p-nitrophenol hydroxylase (p-NPH) activity, a market of CYP2E1, was increased. It is concluded that CDBM may be an inducer of CYP2E1. These results combined with literature data demonstrate that the oxidation of CDBM was catalysed mainly by CYP2E1 and CYP2Bs and that there may be a risk of bromide accumulation following repeated uptake of the trihalomethane.


Management and sustainability of midwifery group practice: Thematic and lexical analyses of midwife interviews.
Although there is high-level evidence supporting positive perinatal outcomes for midwifery group practice (MGP) care, not all women can access this model due to a failure to implement or sustain it. The way that MGPs are managed could be an important factor in whether they are successful in the long-term. To explore what determines optimal management of MGP in Australia, and the influence it has on sustainability of MGP. Interviews were conducted with MGP midwives (n=8). Transcriptions of the audio recordings were analysed thematically and lexically for triangulation. Following a thematic analysis of the data, an over-arching theme emerged - namely - being valued and supported, with three sub-themes: a nice little sweet spot, someone to stand up for you, and building relationships and support beyond the model. This revealed that value and support by all stakeholders, including the midwives themselves, was necessary to sustain an MGP. The lexical analysis revealed the themes: support, the system, and the caseload. This analysis highlighted the importance of the system - specifically, the impact of the institution on how MGP is operationalised. For MGP to be sustained in Australia, it needs to be prioritised, nurtured, and embraced by the whole maternity care system. Management plays an important role in ensuring the right people are employed to pivotal positions and that midwifery-led models of care are valued and supported throughout the system.


Projected incremental changes to extreme wind-driven wave heights for the twenty-first century.
Global climate change will alter wind sea and swell waves, modifying the severity, frequency and impact of episodic coastal flooding and morphological change. Global-scale estimates of increases to coastal impacts have been typically attributed to sea level rise and not specifically to changes to waves on their own. This study provides a reduced complexity method for applying projected extreme wave changes to local scale impact studies. We use non-stationary extreme value analysis to distil an incremental change signal in extreme wave heights and associate this with a change in the frequency of events globally. Extreme wave heights are not projected to increase everywhere. We find that the largest increases will typically be experienced at higher latitudes, and that there is high ensemble model agreement on an increase (doubling of events) for the waters south of Australia, the Arabian Sea and the Gulf of Guinea by the end of the twenty-first century.


Renal vein and vena caval thrombus: a rare presentation of emphysematous pyelonephritis.
Emphysematous pyelonephritis (EPN) can present with symptoms due to infection or sepsis. EPN presenting with renal vein and inferior vena cava (IVC) thrombus is very rare. We report a case of EPN with renal vein and IVC thrombus and its management.


End-tidal carbon dioxide (ETCO2) at intubation and its increase after 10 minutes resuscitation predicts survival with good neurological outcome in out-of-hospital cardiac arrest patients.
To evaluate whether end-tidal carbon dioxide (ETCO2) value at intubation and its early increase (10 min) after intubation predict both the survival to hospital admission and the survival at hospital discharge, including good neurological outcome (CPC 1-2), in patients with out-of-hospital cardiac arrest (OHCA). All consecutive OHCA patients of any etiology between 2015 and 2018 in Pavia Province (Italy) and Ticino Region (Switzerland) were considered. Patients died before ambulance arrival, with a "do-not-resuscitate" order, without ETCO2 value or with incomplete data were excluded. The study population consisted of 668 patients. An ETCO2 value at intubation > 20 mmHg and its increase 10 min after intubation were independent predictors (after correction for known predictors of OHCA outcome) of survival to hospital admission and survival at hospital discharge. Relative to hospital discharge with good neurological outcome, ETCO2 at intubation and its 10-min change were confirmed predictors both individually and in a bivariable analysis (OR 1.83, 95 %CI 1.02-3.3; p = 0.04 and OR 3.9, 95 %CI 1.97-7.74; p < 0.001, respectively). This was confirmed also when accounting for gender, age, etiology and location. After further adjustment for bystander and CPR status, presenting rhythm and EMS arrival time, the ETCO2 change remained an independent predictor. ETCO2 value > 20 mmHg at intubation and its increase during resuscitation improve the prediction of survival at hospital discharge with good neurological outcome of OHCA patients. ETCO2 increase during resuscitation is a more powerful predictor than ETCO2 at intubation. A larger prospective study to confirm this finding appears warranted.


Postharvest application of brassica meal-derived allyl-isothiocyanate to kiwifruit: effect on fruit quality, nutraceutical parameters and physiological response.
The use of natural compounds to preserve fruit quality and develop high value functional products deserves attention especially in the growing industry of processing and packaging ready-to-eat fresh-cut fruit. In this work, potential mechanisms underlying the effects of postharvest biofumigation with brassica meal-derived allyl-isothiocyanate on the physiological responses and quality of 'Hayward' kiwifruits were studied. Fruits were treated with 0.15 mg L-1 of allyl-isothiocyanate vapours for 5 h and then stored in controlled atmosphere (2% O2, 4.5% CO2) at 0 °C and 95% relative humidity, maintaining an ethylene concentration <0.02 μL L-1. The short- and long-term effects of allyl-isothiocyanate on fruit quality traits, nutraceutical attributes, glutathione content, antiradical capacity and the activity of antioxidant enzymes were investigated. The treatment did not influence the overall fruit quality after 120 days of storage, but interestingly it enhanced the ascorbic acid, polyphenols and flavan-3-ol content, improving the antioxidant potential of kiwifruit. The short-term effect of allyl-isothiocyanate was evidenced by an increase of superoxide dismutase activity and of oxidative glutathione redox state, which were restored 24 h after the treatment. The expression levels of genes involved in detoxification functions, ethylene, ascorbate and phenylpropanoid biosynthesis, were also significantly affected upon allyl-isothiocyanate application. These results suggest that allyl-isothiocyanate treatment probably triggered an initial oxidative burst, followed by an induction of protective mechanisms, which finally increased the nutraceutical and technological value of treated kiwifruits.


Evaluation of estimations in vitro of tissue/blood distribution coefficients for organothiophosphate insecticides.
Physiologically based pharmacokinetic (PBPK) modeling of foreign chemicals is dependent on the accurate determination of their tissue/blood distribution coefficients, Kp (partition coefficients). The present study was undertaken to evaluate the validity of the in vitro estimation of the Kp values of the organothiophosphate insecticides parathion and methyl parathion by equilibrium dialysis. Data derived from previously published studies that utilized single-pass perfusions of mouse livers in situ with parathion or methyl parathion were analyzed to determine liver/perfusate Kp values from the equation Kp = (t 1/2ss) (Q)/(0.693) (VH), where Kp is the liver/perfusate distribution ratio, t 1/2ss is the half-life for approach to steady state of the chemical, VH is the liver volume, and Q is the perfusate flow rate. Kp values for methyl parathion were calculated to be 16.4 +/- 7.5 and 9.5 +/- 2.7 (mean +/- SD) for perfused livers and equilibrium dialysis, respectively, while estimates of Kp for parathion were 15.6 +/- 6.3 and 19.5 +/- 5.5 for perfused livers and equilibrium dialysis, respectively. These results indicate that equilibrium dialysis can be utilized to give an accurate estimate of tissue partitioning of parathion and methyl parathion from perfusate into perfused mouse livers.


Author Correction: Proteome analysis reveals a role of rainbow trout lymphoid organs during Yersinia ruckeri infection process.
A correction to this article has been published and is linked from the HTML and PDF versions of this paper. The error has been fixed in the paper.


Cardiac echinococcosis associated with cerebrovascular occlusive disease and subcutaneous bullous eruptions and ulcers.
Cardiac involvement is an uncommon complication of echinococcosis. It is usually asymptomatic and may only be diagnosed incidentally. A 7-year-old boy was admitted with acute stroke and bullous and ulcerated skin lesions. He was diagnosed with cardiac echinococcosis complicated by systemic emboli to the central nervous system and superficial cutaneous arteries. In endemic areas, echinococcosis should be considered in the differential diagnosis of cardiac disease and unexplained cerebral embolism.


Intrahypothalamic noradrenaline injection in the rat enhances operant licking but not lever pressing for milk reward.
Injection of noradrenaline into the hypothalamus of the rat produces poor performance of lever pressing for liquid food reinforcement at moderate ratio values, even though such injection facilitates intake of liquid food available without an operant requirement. The present work shows that a tube-licking operant response was more facilitated and less suppressed by noradrenaline than a lever-pressing response. Noradrenaline produced rear-limb ataxia which might have affected lever pressing more than licking, but results of manipulating the height of the tube suggested that this was unlikely. It is concluded that the operant performance deficit is more evident with lever pressing than with a response naturally involved in ingestion.


Calculated parenteral initial treatment of bacterial infections: Bone and joint infections.
This is the 10th chapter of the guideline "Calculated initial parenteral treatment of bacterial infections in adults - update 2018" in the 2nd updated version. The German guideline by the Paul-Ehrlich-Gesellschaft für Chemotherapie e.V. (PEG) has been translated to address an international audience. This chapter deals with bacterial Infections of bones, joints and prosthetic joints. One of the most pressing points is that after an initial empirical therapy a targeted antimicrobial which penetrates well to the point of infection and is tolerated well over the usually long duration of the therapy is chosen.


The 808 nm and 980 nm infrared laser irradiation affects spore germination and stored calcium homeostasis: A comparative study using delivery hand-pieces with standard (Gaussian) or flat-top profile.
Photobiomodulation relies on the transfer of energy from incident photons to a cell photoacceptor. For many years the concept of photobiomodulation and its outcome has been based upon a belief that the sole receptor within the cell was the mitochondrion. Recently, it has become apparent that there are other photoacceptors operating in different regions of the electromagnetic spectrum. Alternative photoacceptors would appear to be water and mechanisms regulating calcium homeostasis, despite a direct effect of laser photonic energy on intracellular calcium concentration outwith mitochondrial activity or influence, have not been clearly demonstrated. Therefore, to increase the knowledge of intracellular‑calcium and laser photon interaction, as well as to demonstrate differences in irradiation profiles with modern hand-pieces, we tested and compared the photobiomodulatory effect of 808 nm and 980 nm diode laser light by low- and higher-energy (60s, 100 mW/cm2, 100 mW/cm2, 500 mW/cm2, 1000 mW/cm2, 1500 mW/cm2, 2000 mW/cm2) irradiated with a "standard" (Gaussian fluence distribution) hand-piece or with a "flat-top" (uniform fluence) hand-piece. For this purpose, we used the eukaryote unicellular-model Dictyostelium discoideum. The 808 nm and 980 nm infrared laser light, at the energy tested directly affect the stored Ca2+ homeostasis, independent of the mitochondrial respiratory chain activities. From an organism perspective, the effect on Ca2+-dependent signal transduction as the regulator of spore germination in Dictyostelium, demonstrates how a cell can respond quickly to the correct laser photonic stimulus through a different cellular pathway than the known light-chromophore(mitochondria) interaction. Additionally, both hand-piece designs tested were able to photobiomodulate the D. discoideum cell; however, the hand-piece with a flat-top profile, through uniform fluence levels allows more effective and reproducible effects.


Rheumatoid factor in acute bacterial endocarditis.
Sera from 55 parenteral drug abusers with endocarditis due to Staphylococcus aureus were assayed for the presence and titer of rheumatoid factor. Thirteen (24%) of the 55 patients with endocarditis had sera positive for rheumatoid factor at one point or another in their courses; only 2 (7%) of 30 noninfected drug users were found to be positive. It appeared that more severe cases, as evidenced by duration of fever after initiation of antibiotic therapy, were more likely to develop rheumatoid factor.


Brain networks, dimensionality, and global signal averaging in resting-state fMRI: Hierarchical network structure results in low-dimensional spatiotemporal dynamics.
One of the most controversial practices in resting-state fMRI functional connectivity studies is whether or not to regress out the global average brain signal (GS) during artifact removal. Some groups have argued that it is absolutely essential to regress out the GS in order to fully remove head motion, respiration, and other global imaging artifacts. Others have argued that removing the GS distorts the resulting correlation matrices and inappropriately alters the results of group comparisons and relationships to behavior. At the core of this argument is the assessment of dimensionality in terms of the number of brain networks with uncorrelated time series. If the dimensionality is high, then the distortions due to GS removal could be effectively negligible. In the current paper, we examine the dimensionality of resting-state fMRI data using principal component analyses (PCA) and network clustering analyses. In two independent datasets (Set 1: N = 62, Set 2: N = 32), scree plots of the eigenvalues level off at or prior to 10 principal components, with prominent elbows at 3 and 7 components. While network clustering analyses have previously demonstrated that numerous networks can be distinguished with high thresholding of the voxel-wise correlation matrices, lower thresholding reveals a lower-dimensional hierarchical structure, with the first prominent branch at 2 networks (corresponding to the previously described "task-positive"/"task-negative" distinction) and further stable subdivisions at 4, 7 and 17. Since inter-correlated time series within these larger branches do not cancel to zero when averaged, the hierarchical nature of the correlation structure results in low effective dimensionality. Consistent with this, partial correlation analyses revealed that network-specific variance remains present in the GS at each level of the hierarchy, accounting for at least 14-18% of the overall GS variance in each dataset. These results demonstrate that GS regression is expected to remove substantial portions of network-specific brain signals along with artifacts, not simply whole-brain signals corresponding to arousal levels. We highlight alternative means of controlling for residual global artifacts when not removing the GS.


Abnormal expression pattern of lncRNA H19 participates in multiple myeloma bone disease by unbalancing osteogenesis and osteolysis.
Accumulating genetic and epigenetic alterations in multiple myeloma (MM) have been demonstrated to be closely associated with osteolytic bone disease, generally characterized as increased osteoclast formation and decreased osteoblast activity. Previously, serum long non-coding RNA (lncRNA) H19 has been proved to be a biomarker for the diagnosis of MM. Whereas, its role in MM-associated bone homeostasis remains largely elusive. A cohort of 42 MM patients and 40 healthy volunteers were enrolled for evaluating differential expressions of H19 and its downstream effectors. The proliferative capacity of MM cells was monitored by CCK-8 assay. Alkaline phosphatase (ALP) staining and activity detection, either with Alizarin red staining (ARS) were employed to assess osteoblast formation. Osteoblast- or osteoclast-associated gene were detected using qRT-PCR and western blot analysis. Bioinformatics analysis, RNA pull-down, RNA immunoprecipitation (RIP), and chromatin immunoprecipitation (ChIP) were subjected to verify H19/miR-532-3p/E2F7/EZH2 axis, which was accounted for epigenetic suppression of PTEN. The functional role of H19 on MM development through unbalancing osteolysis and osteogenesis was also confirmed in the murine MM model. Upregulation of serum H19 was observed in MM patients, suggesting its positive correlation with the poor prognosis of MM patients. Loss of H19 dramatically weakened cell proliferation of MM cells, promoted osteoblastic differentiation, and impaired osteoclast activity. While reinforced H19 exhibited the opposite effects. Akt/mTOR signaling plays an indispensable role in H19-mediated osteoblast formation and osteoclastgenesis. Mechanistically, H19 served as a sponge for miR-532-3p to upregulate E2F7, a transcriptional activator of EZH2, thereby accounting for modulating epigenetic suppression of PTEN. The in vivo experiments further validated that H19 exerted important impacts on tumor growth through breaking the balance between osteogenesis and osteolysis via Akt/mTOR signaling. Collectively, increased enrichment of H19 in MM cells exhibits an essential role in MM development by disturbing bone homeostasis.


Use of matrix-assisted laser desorption/ionization time-of-flight mass spectrometry for multiplex genotyping.
After completion of the human genome project, the focus of geneticists has shifted to elucidation of gene function and genetic diversity to understand the mechanisms of complex diseases or variation of patient response in drug treatment. In the past decade, many different genotyping techniques have been described for the detection of single-nucleotide polymorphisms (SNPs) and other common polymorphic variants. Matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF-MS) is among the most powerful and widely used genotyping technologies. The method offers great flexibility in assay design and enables highly accurate genotyping at high sample throughput. Different strategies for allele discrimination and quantification have been combined with MALDI (hybridization, ligation, cleavage, and primer extension). Approaches based on primer extension have become the most popular applications. This combination enables rapid and reliable multiplexing of SNPs and other common variants, and makes MALDI-TOF-MS well suited for large-scale studies in fine-mapping and verification of genome-wide scans. In contrast to standard genotyping, more demanding approaches have enabled genotyping of DNA pools, molecular haplotyping or the detection of free circulating DNA for prenatal or cancer diagnostics. In addition, MALDI can also be used in novel applications as DNA methylation analysis, expression profiling, and resequencing. This review gives an introduction to multiplex genotyping by MALDI-MS and will focus on the latest developments of this technology.


Molecular cloning, bioinformatics analysis and functional characterization of B-cell activating factor in goat (Capra hircus).
B-cell activating factor of the TNF family (BAFF) induces B cell survival, proliferation, immunoglobulin secretion and has a role in enhancing immune responses. In the present study, we amplified the cDNA of goat (Capra hircus) BAFF (designated gBAFF) from spleen by reverse transcription-PCR (RT-PCR). The open reading frame (ORF) of gBAFF covers 843 bp encoding 280 amino acids, with a 152-aa mature peptide. Sequence comparison indicated that the amino acid of gBAFF possessed the TNF signature, a transmembrane domain, a putative furin protease cleavage site and three cysteine residues. The predicted three-dimensional (3D) structure of the soluble part of gBAFF (gsBAFF) analyzed by "comparative protein modelling" revealed that it was very similar to its counterparts. Real-time quantitative PCR (qPCR) analysis indicated that gBAFF mRNA was predominantly expressed in goat lymphoid tissue spleen. Recombinant gsBAFF was fused with a small ubiquitin-related modifier (SUMO) gene to enhance the soluble expression level in Escherichia coli BL21 (DE3). The resulting fused protein SUMO-gsBAFF was efficiently expressed and purified using metal chelate affinity chromatography (Ni-NTA), then confirmed by SDS-PAGE and Western blotting analysis. In vitro, the MTT assays and flow cytometric analysis indicated that SUMO-gsBAFF as well as gsBAFF could promote the survival/proliferation of goat splenic B cells or mouse splenic B cells. Therefore, BAFF may be a potential immunologic factor for enhancing immunological efficacy in goat.


A self-assessment tool was reliable in identifying hazards in the homes of elders.
Falls are a leading cause of fatal and nonfatal injuries, particularly among the elderly. A reliable instrument for self-assessment of home falls hazards would facilitate screening for falls risk. This study examined the reliability of self-assessment of home falls hazards by elderly women. Participants were 52 elderly women, aged 67 to 97. All evaluations were performed in the participants' homes. Home falls hazards were evaluated independently by study participants and by trained observers. Kappa statistics indicated good to excellent agreement for most of the environmental factors. However, observers were significantly more likely than the study participants to report certain tripping hazards, particularly objects in walkways. This home checklist is an important step towards a reliable self-report instrument for measuring home falls hazards. Self-assessment appears to be a reliable method for assessing many putative hazards of falling in the home. However, our findings raise questions regarding the reliable assessment of tripping hazards.


Ineffective implantable cardioverter-defibrillator shocks among patients on continuous left ventricular assist device support: Clinical characteristics and management.
Placement of a left ventricular assist device (LVAD) has been described to compromise implantable cardioverter-defibrillator (ICD) defibrillation threshold (DFT). Elevated DFT will have negative consequences and increases the risk of ineffective ICD shocks, morbidity, and mortality. DFT testing is not routinely performed in clinical practice, despite this fact. We describe the clinical characteristics of 7 LVAD patients who presented with multiple ineffective ICD shocks, along with the management strategy in such patients. Seven patients (5 male, mean age 52.2 ± 9 years, 85.7% nonischemic cardiomyopathy) with an ICD in situ who progressed to NYHA class IV, ACC/AHA stage D chronic systolic congestive heart failure who underwent successful LVAD implantation presented to our institution in the setting of ventricular tachyarrhythmia and ineffective ICD shocks. Six patients underwent implantation of azygos and subclavian coils with subsequent DFT testing. The remaining patient was made comfort care. Five patients had successful DFT testing with azygos (n = 4) and subclavian (n = 1) defibrillation coil implantation. One patient had unsuccessful DFT testing despite evaluation of multiple shock vectors. There were no major or minor vascular complications in any of the cases. There were no procedural-related deaths. This case series highlights the need for a systematic approach to management of ICDs and DFT testing in LVAD patients. The addition of new shock vectors with azygos and subclavian coil implantation may reduce DFT, shock burden, morbidity, and mortality.


Patterns of convulsive susceptibility in the long-sleep and short-sleep selected mouse lines.
It has been hypothesized that the Long-Sleep and Short-Sleep mouse lines were bidirectionally selected for high and low brain excitability, and further, that these differences are mediated by the benzodiazepine/gamma-aminobutyric acid (GABA) receptor-chloride channel complex. Hence, mice from both lines were administered seven convulsants (bicuculline, pentylenetetrazol, 3-carbomethoxy-beta-carboline, picrotoxin, caffeine, flurothyl and strychnine) and myoclonic and clonic seizure latencies recorded. Supporting the original hypothesis, the results show that the two lines were differentiated by all of the convulsants and that in response to the drugs, three distinct convulsive patterns were found. Nevertheless, a simple genetic model accounting for these results was not evident. To further clarify these susceptibility patterns, a convulsant representing each of these patterns (bicuculline, pentylenetetrazol or caffeine) was administered in conjunction with the anticonvulsant-barbiturate phenobarbital or the benzodiazepine antagonist Ro 15-1788. Irrespective of the convulsant given, phenobarbital attenuated both myoclonus and clonus subsequent to all convulsants, while Ro 15-1788 had a more discrete anticonvulsant profile.


Coronary artery aneurysm with thrombus evaluated by multi-detector row computed tomography.
This is a case report of a 60-year-old woman with acute myocardial infarction (AMI) who had coronary aneurysms and thrombus in the right coronary artery detected by multi-detector row computed tomography (MDCT). In this case, MDCT was useful for clarifying the thrombus complicated with coronary aneurysms, which was considered to be the cause of AMI. Thus, warfarin was started as an anticoagulant therapy for thrombus.


Repeated quantitative angiograms in coronary arterial aneurysm in Kawasaki disease.
Accurate evaluation of the extent of coronary artery lesions complicating Kawasaki disease is clinically important in patient management. Based on a total of 188 coronary angiograms and retrospective follow-up observations, the condition of coronary aneurysms was quantitatively graded as: 0 (normal)--no significant enlargement in any portion of the coronary artery; I (mild)--aneurysmal dilatation of the coronary artery evident but localized, with a maximal diameter of less than 4.0 mm; II (moderate)--maximal diameter of coronary aneurysms between 4.0 and 8.0 mm, regardless of body size; III (severe)--giant aneurysms, with the maximal diameter greater than 8.0 mm. Most mild coronary aneurysms regressed to normal within a short time, and the patient's prognosis was good. The course of grade II aneurysms varied, depending on initial angiographic coronary diameter, but all were eventually reduced in coronary size. In contrast, grade III aneurysms usually progressed to become obstructive or stenotic coronary lesions, or the large aneurysm persisted. Follow-up observations revealed that the course of coronary artery disease depended on the size and distribution of aneurysms at initial angiography. This grading of the severity of coronary lesions may provide useful criteria for predicting the prognosis of patients with Kawasaki disease.


Alu-minating the Mechanisms Underlying Primate Cortex Evolution.
The higher-order cognitive functions observed in primates correlate with the evolutionary enhancement of cortical volume and folding, which in turn are driven by the primate-specific expansion of cellular diversity in the developing cortex. Underlying these changes is the diversification of molecular features including the creation of human and/or primate-specific genes, the activation of specific molecular pathways, and the interplay of diverse layers of gene regulation. We review and discuss evidence for connections between Alu elements and primate brain evolution, the evolutionary milestones of which are known to coincide along primate lineages. Alus are repetitive elements that contribute extensively to the acquisition of novel genes and the expansion of diverse gene regulatory layers, including enhancers, alternative splicing, RNA editing, and microRNA pathways. By reviewing the impact of Alus on molecular features linked to cortical expansions or gyrification or implications in cognitive deficits, we suggest that future research focusing on the role of Alu-derived molecular events in the context of brain development may greatly advance our understanding of higher-order cognitive functions and neurologic disorders.


The use of anakinra in the treatment of secondary hemophagocytic lymphohistiocytosis.
Hemophagocytic lymphohistiocytosis (HLH) can be familial or secondary, which is often triggered by infection or malignancy. HLH therapy includes dexamethasone and etoposide. However, therapy is associated with significant morbidity and mortality. Anakinra, a recombinant interleukin-1 receptor antagonist, has been reported to treat macrophage activation syndrome (MAS), rheumatic sHLH. We report our experience with anakinra to treat patients with nonrheumatic secondary HLH (sHLH). Six children were diagnosed with HLH from December 2014 to August 2016 and were treated with subcutaneous anakinra (6-10 mg/kg/day divided over four doses) with or without dexamethasone (10 mg/m2 /day). Therapy was either escalated or weaned based on clinical and laboratory response. Five of six patients were treated with anakinra and dexamethasone, and one with anakinra alone due to active cytomegalovirus (CMV) pneumonitis. The median age of diagnosis was 1.8 years (range 0.8-14.9 years). No pathogenic mutations associated with HLH were identified, but three of six possessed genetic variants of unknown significance. Infectious triggers were identified for four patients and two patients had malignancies. The average treatment duration was 8 weeks with 3.5-5.5 years of follow up. No patient needed escalation of therapy to include etoposide. All patients achieved remission. Anakinra was well tolerated without significant adverse effects. Initial treatment with anakinra (with or without dexamethasone) is a feasible treatment alternative for patients with secondary HLH and may allow for avoidance of etoposide. We recommend early initiation of anakinra when HLH is suspected. A broader investigation of the use of anakinra as a first-line agent for HLH is ongoing.


Inhibiting of circ-TLK1 inhibits the progression of glioma through down-regulating PANX1 via targeting miR-17-5p.
Glioma remains the most common malignant tumors in the central nervous system and often has poor prognosis. In recent years, it has been gradually revealed that non-coding RNA effects glioma progression. In this study, we aimed to investigate the significance of circular RNA TLK1 (Circ-TLK1) in predicting the survival of glioma patients as well as its role in glioma development via both in-vitro and in-vivo experiments. We found that Circ-TLK1 was conspicuously up-regulated in glioma tissues compared with adjacent normal tissues, and the up-regulated Circ-TLK1 was significantly correlated with glioma patients' larger tumor volume and higher grades. Functionally, Circ-TLK1 over-expression facilitated glioma growth, migration and invasion, inhibited cell apoptosis, and accelerated PANX1/MAPK/ERK expression, while Circ-TLK1 low expression had the opposite effects. In addition, bioinformatics analysis showed that miR-17-5p was a potential target of Circ-TLK1 and targeted at PANX1. Furthermore, through dual luciferase viability assay, Circ-TLK1 acted as a competing endogenous RNA by sponging miR-17-5p, which targeted and inhibited PANX1/MAPK/ERK expression. MiR-17-5p overexpression mitigated glioma progression, which was significantly inhibited with Circ-TLK1 upregulation. In conclusion, this study confirmed a novel axis of Circ-TLK1-miR-17-5p-PANX1 in modulating glioma development, providing more references for glioma diagnosis and targeted therapy.


Five metastasis-related mRNAs signature predicting the survival of patients with liver hepatocellular carcinoma.
Liver hepatocellular carcinoma (HCC) is one of the most malignant tumors, of which prognosis is unsatisfactory in most cases and metastatic of HCC often results in poor prognosis. In this study, we aimed to construct a metastasis- related mRNAs prognostic model to increase the accuracy of prediction of HCC prognosis. Three hundred seventy-four HCC samples and 50 normal samples were downloaded from The Cancer Genome Atlas (TCGA) database, involving transcriptomic and clinical data. Metastatic-related genes were acquired from HCMBD website at the same time. Two hundred thirty-three samples were randomly divided into train dataset and test dataset with a proportion of 1:1 by using caret package in R. Kaplan-Meier method and univariate Cox regression analysis and lasso regression analysis were performed to obtain metastasis-related mRNAs which played significant roles in prognosis. Then, using multivariate Cox regression analysis, a prognostic prediction model was established. Transcriptome and clinical data were combined to construct a prognostic model and a nomogram for OS evaluation. Functional enrichment in high- and low-risk groups were also analyzed by GSEA. An entire set based on The International Cancer Genome Consortium(ICGC) database was also applied to verify the model. The expression levels of SLC2A1, CDCA8, ATG10 and HOXD9 are higher in tumor samples and lower in normal tissue samples. The expression of TPM1 in clinical sample tissues is just the opposite. One thousand eight hundred ninety-five metastasis-related mRNAs were screened and 6 mRNAs were associated with prognosis. The overall survival (OS)-related prognostic model based on 5 MRGs (TPM1,SLC2A1, CDCA8, ATG10 and HOXD9) was significantly stratified HCC patients into high- and low-risk groups. The AUC values of the 5-gene prognostic signature at 1 year, 2 years, and 3 years were 0.786,0.786 and 0.777. A risk score based on the signature was a significantly independent prognostic factor (HR = 1.434; 95%CI = 1.275-1.612; P < 0.001) for HCC patients. A nomogram which incorporated the 5-gene signature and clinical features was also built for prognostic prediction. GSEA results that low- and high-risk group had an obviously difference in part of pathways. The value of this model was validated in test dataset and ICGC database. Metastasis-related mRNAs prognostic model was verified that it had a predictable value on the prognosis of HCC, which could be helpful for gene targeted therapy.


Val50Met hereditary transthyretin amyloidosis: not just a medical problem, but a psychosocial burden.
Hereditary transthyretin (TTR) amyloidosis (ATTRv) is a heterogeneous disease with a clinical presentation that varies according to geographical area and TTR mutation. The symptoms of Val50Met-ATTRv are mainly neuropathic and progress to complete disability and death in most untreated patients within 10 to 15 years of diagnosis. The neurological effects may also be accompanied by gastrointestinal impairment, cardiomyopathy, nephropathy and/or ocular deposition. The disease is thus associated with a high degree of patient disability. Accordingly, we aimed to describe the psychosocial burden associated with ATTRv in a group of patients, asymptomatic Val50Met carriers, relatives and caregivers in the endemic focus of the disease in Majorca via a survey addressing various aspects related to psychosocial burden. We performed a an observational, descriptive, cross-sectional and multicentre study in order to analyze the prevalence of self-reported impact of ATTRv disease upon their daily life. In addition to the self-knowledge, fear and burden related to the disease. The survey was disseminated during the regular follow up at the outpatient clinic of the Hospital Universitario Son Llàtzer and during the meetings organized by the Andrade's Disease patients' advocacy group from the Balearic Islands. These meetings were attended also by subjects followed up by the Hospital Universitario Son Espases and their caregivers and relatives. Survey was self-administrated. No intervention was done by the investigators. 85 subjects completed the survey: 61 carrying the TTR-V50M variant and 24 caregivers or relatives. Our study revealed that, although most of the population studied had had prior contact with ATTRv through affected relatives, there was still a lack of information regarding disease diagnosis. Fear of the genetic test result and psychological issues were common in our population. Moreover, the disease had a stronger impact on the daily life of our patients than that of our asymptomatic carriers. Autonomic symptoms were the main source of burden for relatives and caregivers. Our survey results show high psychosocial burden associated with Val50Met-ATTRv in our area.


Tracing the wider impacts of biomedical research: a literature search to develop a novel citation categorisation technique.
There is an increasing need both to understand the translation of biomedical research into improved healthcare and to assess the range of wider impacts from health research such as improved health policies, health practices and healthcare. Conducting such assessments is complex and new methods are being sought. Our new approach involves several steps. First, we developed a qualitative citation analysis technique to apply to biomedical research in order to assess the contribution that individual papers made to further research. Second, using this method, we then proposed to trace the citations to the original research through a series of generations of citing papers. Third, we aimed eventually to assess the wider impacts of the various generations. This article describes our comprehensive literature search to inform the new technique. We searched various databases, specific bibliometrics journals and the bibliographies of key papers. After excluding irrelevant papers we reviewed those remaining for either general or specific details that could inform development of our new technique. Various characteristics of citations were identified that had been found to predict their importance to the citing paper including the citation's location; number of citation occasions and whether the author(s) of the cited paper were named within the citing paper. We combined these objective characteristics with subjective approaches also identified from the literature search to develop a citation categorisation technique that would allow us to achieve the first of the steps above, i.e., being able routinely to assess the contribution that individual papers make to further research.


Zebrafish eggs used as bioreactors for the production of bioactive tilapia insulin-like growth factors.
Multiple advantages-including the short generation time, large numbers of fertilized eggs, low cost of cultivation and easy maintenance favor the use of fish as bioreactors for the production of pharmaceutical proteins. In the present study, zebrafish eggs were used as bioreactors to produce mature tilapia insulin-like growth factors (IGFs) proteins using the oocyte-specific zona pellucida (zp3) promoter. The chimeric expression plasmids, pT2-ZP-tIGFs-IRES-hrGFP, in which hrGFP was used as reporter of tilapia IGFs expression, were designed to established Tg (ZP:tIGFs:hrGFP) transgenic lines for the expression of tilapia IGF-1 and IGF-2. Recombinant tilapia IGF-1 and IGF-2 were expressed as soluble forms in cytoplasm of fertilized eggs. The content level of tilapia IGF-1 and IGF-2 were 6.5 and 5.0% of the soluble protein, respectively. Using a simple Ni-NTA affinity chromatography purification process, 0.58 and 0.49 mg of purified tilapia IGF-1 and IGF-2 were obtained, respectively, from 650 fertilized eggs. The biological activity of the purified tilapia IGF-1 and IGF-2 was confirmed via a colorimetric bioassay to monitor the growth stimulation of zebrafish embryonic cells (ZF4), tilapia ovary cells (TO-2) and human osteosarcoma epithelial cells (U2OS). These results demonstrate that the use of zebrafish eggs as bioreactors is a promising approach for the production of biological recombinant proteins.


Sequence-Specific Control of Mitochondrial Gene Transcription Using Programmable Synthetic Gene Switches Called MITO-PIPs.
Mitochondria possess multiple copies of mitochondrial DNA (mtDNA) that encode 37 genes and their transcription and replication get controlled by unique molecular codes different from that in the nuclear DNA. The mtDNA has been gaining increased attention as one of the critical therapeutic targets as mutations in them impair the function of mitochondria and cause mitochondrial diseases like MELAS. In this chapter, we describe artificial control of mitochondrial transcription based on mtDNA sequence information with a new type of compounds termed MITO-PIPs, which encompasses two domains: pyrrole-imidazole polyamide as DNA recognition domain and mitochondrial penetrating peptide as the mitochondria-targeting domain. Because MITO-PIPs are amenable to tunability, they can be expanded as a synthetic strategy to modulate mitochondrial gene(s) on demand.


Pharmacokinetics, tolerability, and performance of a novel matrix transdermal delivery system of fentanyl relative to the commercially available reservoir formulation in healthy subjects.
A novel transdermal formulation of fentanyl-containing dipropylene glycol droplets dispersed in a silicone matrix with a rate-controlling membrane was developed. Healthy male subjects (n = 24) received repeated 72-hour applications of fentanyl (50 mug/h) as the novel matrix and the conventional reservoir formulations in a randomized, 2-way crossover study. Blood samples were collected, and serum concentrations of fentanyl were assayed using liquid chromatography with mass spectrometry detection. The mean area under the curve (AUCtau) and peak concentrations (C(max)) of the matrix formulation were 84 838 pg.h/mL and 1680 pg/mL, respectively. Ratio and 90% confidence intervals of AUCtau and C(max) between the 2 formulations were within 80% to 125%. Adherence of the matrix formulation was higher than the reservoir formulation (62.5 vs 56.2%, P < .0001), without affecting skin irritation. Vital signs and adverse events of the 2 formulations were similar in nature and frequency. The novel matrix formulation displayed enhanced adherence and resulted in similar pharmacokinetics and tolerability as the reservoir formulation.


Synthesis and antitumor activity of novel silibinin and 2,3-dehydrosilybin derivatives with carbamate groups.
A novel series of silibinin and 2,3-dehydrosilybin derivatives bearing carbamate groups were designed, synthesized and their in vitro anticancer activities were screened against human cancer cell lines including MCF-7, NCI-H1299, HepG2 and HT29 by CCK-8 assay. The results showed that most of the compounds significantly suppressed the proliferation of tested cancer cells. Among them, compounds 2h, 3h and 3f demonstrated markedly higher antiproliferative activity on MCF-7 cells with IC50 values of 2.08, 5.54 and 6.84 µM, respectively. Compounds 3e, 3g and 2g displayed better cytotoxic activity against NCI-H1299 cells with IC50 values of 8.07, 8.45 and 9.09 µM, respectively. Compounds 3g, 3c and 3h exhibited a promising inhibitory effect against HepG2 cells with IC50 values of 8.88, 9.47 and 9.99 µM, respectively. Compounds 3e, 2e and 3c revealed effective biological potency on HT29 cells with IC50 values of 6.27, 9.13 and 9.32 µM, respectively. In addition, the outcomes of the docking studies between compounds 2f, 2h, 3e, 3g and Hsp90 receptor (PDB ID: 4AWO) suggest the possible mechanism of inhibition against MCF-7 cell lines. Graphical abstract.


Validation of tumour-free distance as novel prognostic marker in early-stage cervical cancer: a retrospective, single-centre, cohort study.
The aim of the present study was to assess the prognostic value of tumour-free distance (TFD), defined as the minimum distance of uninvolved stroma between the tumour and peri-cervical stromal ring, in early-stage cervical cancer. Patients with pathologic FIGO 2009 stage IA1-IIA2 cervical cancer, treated by primary radical surgical treatment between 01/2000 and 11/2019, were retrospectively included. Adjuvant treatment was administered according to the presence of previously established pathologic risk factors. TFD was measured histologically on the hysterectomy specimen. Pre-operative TFD measured at MRI-scan from a cohort of patients was reviewed and compared with pathology TFD. 395 patients were included in the study. 93 (23.5%) patients had TFD ≤ 3.0 mm and 302 (76.5%) had TFD > 3.0 mm. TFD ≤ 3.0 mm together with lymph vascular space involvement represented the strongest predictor for lymph node metastasis at multivariate analysis. TFD ≤ 3.0 mm was associated with worse 5-year disease-free survival (DFS) and overall survival (OS), compared with TFD > 3.0 mm (p = 0.022 and p = 0.008, respectively). DFS difference was more evident in the subgroup of patients with low-risk factors who did not receive adjuvant treatment (p = 0.002). Cohen's kappa demonstrated an agreement between TFD measured at pre-operative MRI-scan and histology of 0.654. Pathologic TFD ≤ 3.0 mm represents a poor prognostic factor significantly associated with lymph node metastasis and it may be considered a novel marker to select candidates for adjuvant treatment. The possibility to obtain this parameter by radiological imaging makes it a potential easy-measurable pre-operative marker to predict the presence of high-risk pathologic factors in early-stage cervical cancer.


A set of 150 pictures with morphologically complex English compound names: norms for name agreement, familiarity, image agreement, and visual complexity.
We present a set of 150 pictures with morphologically complex English compound names. The pictures were collected from various sources and were standardized to appear as grayscale line drawings of a fixed size. All the compounds had two constituents and were primarily of the noun-noun type. Following previous studies, we collected name agreement (percentage and H), familiarity, image agreement, and visual complexity norms, as well as frequency estimates for the whole compound word and its first and second constituents. These pictures and their corresponding norms (available from the Psychonomic Society's supplemental archive) are a valuable tool in the study of the morphological representation of complex words in language processing.


Systemic immune response induced by oxaliplatin-based neoadjuvant therapy favours survival without metastatic progression in high-risk rectal cancer.
Systemic failure remains a challenge in rectal cancer. We investigated the possible systemic anti-tumour immune activity invoked within oxaliplatin-based neoadjuvant therapy. In two high-risk patient cohorts, we assessed the circulating levels of the fms-like tyrosine kinase 3 ligand (Flt3L), a factor reflecting both therapy-induced myelosuppression and activation of tumour antigen-presenting dendritic cells, at baseline and following induction chemotherapy and sequential chemoradiotherapy, both modalities containing oxaliplatin. The primary end point was progression-free survival (PFS). In both cohorts, the median Flt3L level was significantly higher at completion of each sequential modality than at baseline. The 5-year PFS (most events being metastatic progression) was 68% and 71% in the two cohorts consisting of 33% and 52% T4 cases. In the principal cohort, a high Flt3L level following the induction chemotherapy was associated with low risk for a PFS event (HR: 0.15; P < 0.01). These patients also had available dose scheduling and toxicity data, revealing that oxaliplatin dose reduction during chemoradiotherapy, undertaken to maintain compliance to the radiotherapy protocol, was associated with advantageous PFS (HR: 0.47; P = 0.046). In high-risk rectal cancer, oxaliplatin-containing neoadjuvant therapy may promote an immune response that favours survival without metastatic progression.


Carcinoma of the extrahepatic bile ducts.
Thirty-seven patients with carcinoma of the extrahepatic bile ducts (EHBD), diagnosed over 14 years from 1974 to 1987, were studied retrospectively. The most common presenting complaint was jaundice, followed by abdominal pain, nausea and vomiting, and weight loss. The majority of EHBD carcinomas were located at the middle third of the bile duct (43.2%), followed by upper third bile duct (37.8%), and lower third bile duct (18.9%). The resectability of the surgically managed group was 44.4%. The most common histologic type was adenocarcinoma. The 30-day postoperative mortality was 30.6%. The mean overall survival of the patients with EHBD carcinoma was 11.2 months. Patients with distal third tumors had the best prognosis with a mean overall survival of 16.0 months. The mean overall survival of patients with upper-third and middle-third tumors were 10.9 months and 9.4 months. A high index of suspicion of the disease as well as earlier, aggressive surgery may improve patient survival.


Expression of Bcl-2 protein in the epiphyseal plate cartilage and trabecular bone of growing rats.
The protooncogene protein, Bcl-2, protects cells from apoptosis and ensures their survival in vitro by inhibiting the action of the apoptosis-inducer, Bax. Its expression in proliferative and long-lived cells in vivo also indicates that it protects against cell death. The chondrocytes of the epiphyseal plate cartilage undergo a series of maturation steps and deposit mineral in the cartilage matrix before dying. The possibility that Bcl-2 helps protect chondrocytes until mineral deposition is completed was investigated by determining the distribution of Bcl-2 immunoreactivity in the epiphyseal plate cartilage of growing rats and its subcellular localization, using a specific antibody. The involvement of Bax in the triggering of chondrocyte death was checked by immunocytochemistry. Bcl-2 expression in the osteoblasts and the final result of their evolution, the osteocytes, was also examined in trabecular bone. Bcl-2 immunoreactivity was non-uniformly distributed throughout the epiphyseal cartilage. It was maximal in proliferative chondrocytes, decreased in mature chondrocytes, and low in hypertrophic chondrocytes, whereas there was Bax immunoreactivity in all chondrocytes examined. Immunolabeling was intense in osteoblasts but considerably lower in fully differentiated osteocytes. Bcl-2 immunoreactivity was mainly in the cytoplasm of chondrocytes, osteoblasts, and early osteocytes; the nuclei appeared clear. The subcellular distribution of Bcl-2 immunolabeling in chondrocytes, revealed by gold particles in the electron microscope, showed that gold particles were frequently concentrated in the mitochondria in all the cartilage zones and lay mainly within the organelles, not at their periphery. The endoplasmic reticulum contained moderate immunoreactivity and there were few gold particles in the cytoplasm and nuclei. The number of gold particles decreased in all the subcellular compartments from proliferative to hypertrophic chondrocytes. In contrast, Bax immunoreactivity changed little during chondrocyte terminal evolution, and its subcellular distribution mirrored that of Bcl-2. These immunocytochemical data indicate that Bcl-2 helps maintain chondrocytes and osteoblasts until their terminal maturation.


Effects of n-propyl gallate on neuronal survival after forebrain ischemia in rats.
The present study was conducted to assess the effects of intraperitoneal administration of n-propyl gallate (PG) on hippocampal neuronal survival after forebrain ischemia. Forty male Sprague-Dawley rats were randomly assigned to one of 6 groups. Animals in the PG-I-10, PG-I-8 and PG-S groups received intraperitoneal injection of PG (100mg/kg) 72, 48, 24h and 30 min before severe (10 min) or moderate (8 min) ischemia or sham operation, respectively, while animals in the V-I-10, V-I-8 and V-S groups received the vehicle (10% DMSO) in the same manner. Forebrain ischemia was produced by bilateral carotid occlusion combined with hypotension (35 mmHg) under isoflurane anesthesia. Animals were killed 7 days after reperfusion. Histological assessments were performed using hematoxylin and eosin staining. In separate groups of animals that received PG or vehicle, m-RNA levels of hypoxia-inducible factor 1α (HIF-1α), erythropoietin (EPO) and vascular endothelial growth factor (VEGF) were measured using the reverse transcription-PCR protocol. The number of normal neurons was significantly higher in the PG-I-8 group compared with that in the V-I-8 group, whereas it was similar between the PG-I-10 and V-I-10 groups. Animals that received PG had significantly higher levels of HIF-1α, EPO and VEGF expression compared with those that received vehicle. The results indicated that intraperitoneal administration of PG may have neuroprotective effects in a model of moderate, but not severe, forebrain ischemia in rats.


[Management of spondyloarthritis (ESPOGUIA): methodology and general data from the document].
The need for clinical practice guidelines for spondyloarthritis (SpA) comes from the complexity of its management and the great advances that both its treatment and its diagnosis have experiences, which come accompanied by a great variability in its management that exceeds what is considered acceptable. The objective of the ESPOGUIA is to reduce the variability in the management of SA, improving the quality of care through clinical recommendations adapted to their particular environment and based on the best available evidence. For the development of ESPOGUÍA the following methods have been employed: nominal group, systematic review and Delphi surveys. The expert panel is voluntary, with multiple disciplines participating and based on selection criteria. For the first time in a SER recommendation document, the opinion of the patient was included. In a meeting of the nominal group, the reach, objective, users, organization and needs of the systematic review were established. The elaboration of the guideline, along with its recommendations, was performed in parallel to the systematic reviews, with a meeting for agreement held afterward. The scoring of the degree of evidence and the level of the recommendation was established from the evidence levels proposed by the Oxford Center of Evidence-Based Medicine and the degree of agreement was obtained through a two-round Delphi exercise. We have produced a clinical guideline that encompasses all of the SpA, although it has two sub-guidelines, one for ankylosing spondylitis and the other one for psoriatic arthritis. The target users are mainly rheumatologists, although other specialists and paramedical staff, such as nurses or patient associations, might benefit. Its content envelops all of the management aspects of SpA, from clinical suspicion to treatment, follow-up and day-to-day activities of the patient. ESPOGUIA can become a very useful instrument for the management of SA and an interactive reference.


Temperature and moisture requirements for conidial germination of an isolate of Beauveria bassiana, pathogenic to Rhodnius prolixus.
The effects of temperature, relative humidity and water activity on germination of conidia of an isolate of Beauveria bassiana (Bals.) Vuill. pathogenic to the triatomine vector of Chagas' disease, Rhodnius prolixus Stål., were investigated in vitro. Germination occurred at temperatures between 15 degrees C and 35 degrees C under saturated atmosphere and the optima ranged from 25 degrees C to 30 degrees C. At the extreme temperatures tested (15 degrees C and 35 degrees C) the germination process was delayed, but germination rates reached more than 95%. Germination of B. bassiana conidia was strongly affected by moisture conditions. The availability of water, in both atmospheric and liquid conditions, caused changes in germination times as well as in germination rates. For example, at 25 degrees C + 0.5 degrees C, germination took place within 20 h at 95.5% RH, whereas it needed 72 h of incubation at 90% RH. Germination times increased as the water activity declined from 0.96 a(w) to 0.92 a(w). Below 0.92 a(w), no germination was observed after a 72 h incubation time.


The effect of 2-[(aminopropyl)amino] ethanethiol (WR1065) on radiation-induced DNA damage and repair and cell progression in V79 cells.
The radioprotector 2-[(aminopropyl)amino] ethanethiol (WR1065) was investigated with respect to its ability to affect radiation-induced DNA damage and repair in V79 cells. Studies were performed to evaluate the protector under conditions in which it is known to be effective in reducing the cytotoxic and mutagenic effects of gamma-irradiation. At a concentration of 4 mM, WR1065 protected against the formation of single strand breaks (SSB), as determined by the method of alkaline elution, when it was present during irradiation. The protector appeared, however, to inhibit the subsequent postirradiation repair or rejoining of SSB. While repair was complete within 24 h, the protector reduced the rate of repair by a factor of 3. This inhibitory effect on the rate of repair did not correlate with either measured differences in cell survival or mutagenesis. The radioprotector was also investigated with respect to its ability to affect cell cycle progression. WR1065 present in the growth medium inhibited the progression of cells through S-phase, and cell-doubling time following a 3 h exposure to the protector was increased from 11 to 18 h. These data are consistent with the well characterized property of thiols to inhibit DNA polymerase activity. It was concluded that, while the presence of WR1065 during irradiation reduced SSB-DNA damage, its effect on the subsequent rejoining of these breaks could not be correlated with its observed effect on protecting against radiation-induced mutagenesis. It may be that the inhibition of cell-cycle progression by the protector allowed more time to enhance the fidelity of repair as measured by the protector's ability to protect against radiation-induced mutagenesis.


Comparison of methods for the measurement of apoproteins A1 and B in serum.
We evaluated four currently available commercial methods for measuring the concentration of apoproteins A1 and B in human serum (a radioimmunoassay, an immunonephelometric assay, and two electroimmunoassays). We found that the immunonephelometric assay provided the best between-run precision, whereas the two electroimmunoassays gave the poorest. Only the immunoephelometric assay achieved a between-run coefficient of variation of less than 5%, which we believe is a desirable target for this parameter. Comparison with other laboratories was assessed using a reference serum obtained from the Centers for Disease Control, Atlanta, Georgia, USA; we are currently using this as a reference serum employing a range of +/- 15% of its assigned values as acceptable. Linearity of the immunonephelometric assay was satisfactory throughout the ranges of 0.5-2.0 g/L for apoprotein A1, and 0.3-1.8 g/L for apoprotein B.


Dietary very long chain fatty acids directly influence the ratio of tetracosenoic (24:1) to tetracosanoic (24:0) acids of sphingomyelin in rat liver.
Twenty-one groups of weanling male Wistar rats were fed semipurified diets containing 5% (w/w) of different dietary fats. After 2 wk, liver sphingomyelin (SM) fatty acid composition was determined. The ratio of 24:1 to 24:0 in liver SM varied over a tenfold range in response to dietary fat type. Step-wise multiple regression analysis indicated that dietary 24:1, 24:0, and 22:1 were the most significant factors in predicting the 24:1/24:0 ratio of liver SM. The mathematical relation between the dietary fatty acid composition and liver SM 24:1/24:0 was y = 1.88 (24:1) -1.49 (24:0) +0.21 (22:1) +0.01 (18:1) +0.26, r2 = 0.95, P < 0.0001. These results were confirmed by a second experiment in which the rats were fed olive oil-based diets supplemented with various fatty acid ethyl esters.


Subtle genomic DNA damage induces intraneuronal production of amyloid-β (1-42) by increasing β-secretase activity.
Aberrant accumulation of amyloid-β (Aβ) in brain is the major trigger for pathogenesis in Alzheimer's disease (AD). It is imperative to understand how Aβ attains such toxic levels in the brain parenchyma. We detected that a subtle and tolerable amount of DNA damage, related to aging, increased intraneuronal Aβ1-42 production both in cultured neuron and in cortex of rodent brain. Strikingly, we also observed elevated levels of mitochondrial fusion and of its major driver protein, MFN2. Hyperfusion of mitochondria may be seen as an adaptive stress response resulting from the induction of ER stress since we detected the activation of both PERK and IRE1α arms of unfolded protein response of ER stress. We found increased phosphorylation of PERK substrate eukaryotic initiation factor 2 α (eIF2α), and upregulation of the downstream effector proteins, ATF4 and CHOP. Concomitantly, increased XBP1 level, the direct effecter protein of IRE-1α, was observed. Reports suggest that eIF2α phosphorylation can increase BACE1 activity, the rate limiting enzyme in Aβ production. Here, we show that inhibiting PERK, decreased Aβ1-42 level while direct BACE1 inhibition, reduced the mitochondrial fusion. We found increased MFN2 expression in young 5xFAD mice when Aβ plaques and neurodegeneration were absent. Thus, our study indicates that mild DNA damage leads to increased Aβ1-42 production almost as a consequence of an initial ER stress-directed protective mitochondrial fusion in brain. We propose that an age-related subtle genomic DNA damage may trigger enhanced intraneuronal Aβ1-42 production in an apparently healthy neuron way before the appearance of clinical symptoms in AD.


Data on the daily electricity load profile and solar photovoltaic (PV) system components for residential buildings in Lagos, Nigeria.
This article contains the average daily electric load profile (for 24 h of the day) for the five categories of residential buildings (duplex, single family bungalow, traditional court yard, flat/apartment dwelling and 'face-me-I-face-you') in three Local Government Areas (LGAs) of the state of Lagos, Nigeria. In each of the LGAs, 10 buildings per residential building type were surveyed for the collection of data with the aid of a questionnaire. In each surveyed household, a household member completed the energy audit section of the questionnaire with the assistance of the questionnaire administrator while the section of the questionnaire designed as a time-of-use diary was left with the household for completion. For each building surveyed, the data retrieved from the completed time-of-use diary was used in Microsoft Excel for computing the hourly electricity load profile for the seven days of the week. In order to obtain the hourly energy load (in watts) for each building, the power rating of the appliances used during each of the 24 h of the day was summed and the result in watts was converted to kWh by dividing by 1000. Each dwelling's daily load profile was obtained as an average of the load profile for the seven days of the week. The article as well provides data on the solar photovoltaic systems' components designed to supply electricity to the building and the levelized cost of electricity (LCOE) of the systems for the base case scenario and different sensitivity cases obtained from simulations using HOMER Pro. The load profile data provided in this article can be reused by other researchers in the design of solar photovoltaic systems for residential buildings.


Decrease in Statewide Antipsychotic Prescribing after Implementation of Child and Adolescent Psychiatry Consultation Services.
To learn if a quality of care Medicaid child psychiatric consultation service implemented in three different steps was linked to changes in statewide child antipsychotic utilization. Washington State child psychiatry consultation program primary data and Medicaid pharmacy division antipsychotic utilization secondary data from July 1, 2006, through December 31, 2013. Observational study in which consult program data were analyzed with a time series analysis of statewide antipsychotic utilization. All consultation program database information involving antipsychotics was compared to Medicaid pharmacy division database information involving antipsychotic utilization. Washington State's total child Medicaid antipsychotic utilization fell from 0.51 to 0.25 percent. The monthly prevalence of use fell by a mean of 0.022 per thousand per month following the initiation of elective consults (p = .004), by 0.065 following the initiation of age/dose triggered mandatory reviews (p < .001), then by another 0.022 following the initiation of two or more concurrent antipsychotic mandatory reviews (p = .001). High-dose antipsychotic use fell by 57.8 percent in children 6- to 12-year old and fell by 52.1 percent in teens. Statewide antipsychotic prescribing for Medicaid clients fell significantly at different rates following each implementation step of a multilevel consultation and best-practice education service.


Allergens and Adjuvants in Allergen Immunotherapy for Immune Activation, Tolerance, and Resilience.
Allergen immunotherapy (AIT) is the only setting in which a vaccine is applied in patients allergic exactly to the active principle in the vaccine. Therefore, AIT products need to be not only effective but also safe. In Europe, for subcutaneous AIT, this has been achieved by the allergoid strategy in which IgE epitopes are destroyed or masked. In addition, adjuvants physically precipitate the allergen at the injection site to prevent too rapid systemic distribution. The choice of adjuvant critically shapes the efficacy and type of immune response to the injected allergen. In contrast to TH2-promoting adjuvants, others clearly counteract allergy. Marketed products in Europe are formulated with aluminum hydroxide (alum) (66.7%), microcrystalline tyrosine (16.7%), calcium phosphate (11.1%), or the TH1 adjuvant monophosphoryl lipid A (5.6%). In contrast to the European practice, in the United States mostly nonadjuvanted extracts and no allergoids are used for subcutaneous AIT, highlighting not only a regulatory but maybe a "historic preference." Sublingual AIT in the form of drops or tablets is currently applied worldwide without adjuvants, usually with higher safety but lower patient adherence than subcutaneous AIT. This article will discuss how AIT and adjuvants modulate the immune response in the treated patient toward immune activation, modulation, or-with new developments in the pipeline-immune resilience.


Suppression of ICAM-1 in human venous endothelial cells by small interfering RNAs.
Cardiopulmonary bypass-mediated release of proinflammatory cytokines promotes the transendothelial migration of leukocytes. Among others, intercellular adhesion molecule (ICAM) is essential for this migratory process within the venous bypass graft, which finally contributes to a diminished early patency rate by thickening of the intima. Small interfering ribonucleic acids (siRNAs) are efficient and specific modulators of endogenous gene expression. This study describes the application of siRNAs to suppress ICAM-1 expression on the surface of human venous endothelial cells. Primary cultures of human venous endothelial cells were either transfected with ICAM-1 siRNA, with a scrambled control siRNA or cultured without transfection. ICAM-1 expression was analyzed with or without TNF-alpha stimulation by flow cytometry. Upon TNF-alpha stimulation, cells transfected with ICAM-1 siRNA showed a six- to seven-fold decreased ICAM-1 expression compared to untransfected cells or cells transfected with the scrambled control siRNA. This is the first report that ICAM-1 expression can be effectively silenced by siRNAs on endothelial cells from human saphenous veins. This new technology may render novel therapeutic concepts to reduce early graft failure by protecting venous bypass grafts against early intra- or postoperative leukocyte infiltration.


Blood thrombopoietin, IL-6 and IL-11 levels in patients with agnogenic myeloid metaplasia.
Agnogenic myeloid metaplasia (AMM) is a disease characterized by bone marrow megakaryocyte hyperplasia and clusters of megakaryocytes, in which many of the megakaryocytes are atypical. In order to elucidate the mechanisms of megakaryocytosis, ELISA assays of blood levels of thrombopoietin (TPO), interleukin-6 (IL-6) and interleukin-11 (IL-11) were done in 45 patients with AMM and compared with normal volunteer controls. Higher blood TPO levels were found in AMM than in controls (P < 0.0001), and blood TPO levels were correlated with the degree of marrow fibrosis (P = 0.0078). Blood levels of IL-6 were also significantly higher in AMM, when compared with controls (P < 0.0001). However, no correlation was found between blood IL-6 levels and degree of marrow fibrosis. No correlation was found between either TPO or IL-6 and the number of blood platelet counts, the number of marrow megakaryocytes, WBC counts, or the degree of splenomegaly. Blood IL-11 levels were undetectable in most patients and no significant difference was found in AMM as compared to controls. The present study demonstrated that, while in idiopathic thrombocytopenic purpura (ITP) or aplastic anemia, blood TPO levels are relatively correlated with the numbers of platelet and/or megakaryocyte mass, blood TPO levels do not correlate with blood platelet counts, or marrow megakaryocyte mass in AMM. Therefore, in AMM, other mechanisms such as the number of TPO receptors on platelets or megakaryocytes, c-MPL receptor abnormalities, abnormal production of TPO mRNA and so on, will have to be studied. Furthermore, TPO may play a significant role in the pathogenesis of marrow fibrosis; IL-6 may be a factor in the development of marrow megakaryocytosis but its elevated blood levels may represent a secondary immune phenomenon; and IL-11 probably does not play a significant role in causing marrow megakaryocytosis in this disease.


Ionizing radiation increases the endothelial permeability and the transendothelial migration of tumor cells through ADAM10-activation and subsequent degradation of VE-cadherin.
We analyzed the changes in permeability of endothelial cell layers after photon irradiation, with a focus on the metalloproteases ADAM10 and ADAM17, and on VE-cadherin, components crucial for the integrity of endothelial intercellular junctions, and their roles in the transmigration of cancer cells through endothelial cell monolayers. Primary HUVEC were irradiated with 2 or 4 Gy photons at a dose rate of 5 Gy/min. The permeability of an irradiated endothelial monolayer for macromolecules and tumor cells was analyzed in the presence or absence of the ADAM10/17 inhibitors GI254023X and GW280264X. Expression of ADAM10, ADAM17 and VE-Cadherin in endothelial cells was quantified by immunoblotting and qRT. VE-Cadherin was additionally analyzed by immunofluorescence microscopy and ELISA. Ionizing radiation increased the permeability of endothelial monolayers and the transendothelial migration of tumor cells. This was effectively blocked by a selective inhibition (GI254023X) of ADAM10. Irradiation increased both, the expression and activity of ADAM10, which led to increased degradation of VE-cadherin, but also led to higher rates of VE-cadherin internalization. Increased degradation of VE-cadherin was also observed when endothelial monolayers were exposed to tumor-cell conditioned medium, similar to when exposed to recombinant VEGF. Our results suggest a mechanism of irradiation-induced increased permeability and transendothelial migration of tumor cells based on the activation of ADAM10 and the subsequent change of endothelial permeability through the degradation and internalization of VE-cadherin.