In Vitro Cellular & Developmental Biology

In vitro cloning via direct somatic embryogenesis and genetic stability assessment of Paphiopedilum niveum (Rchb.f.) Stein: the endangered Venus's slipper orchid Abstract Wild populations of Paphiopedilum niveum (Rchb.f.) Stein, an endangered orchid species, have been threatened with extinction due to climate change and overcollection. Conventional methods of propagation are unable to meet market demands. The current study reports the production of genetically stable, protocorm-derived protocorm-like bodies (PLBs) of P. niveum that can be used for commercial production. These PLBs are referred to hereafter as somatic embryos (SEs). Factors affecting SE induction and SE proliferation were examined. The highest percentage of SE formation (68.33 ± 11.77%) and the maximum number of SEs per explant (5.19 ± 0.67) were obtained on modified Vacin and Went (MVW) medium containing 0.1 mg L−1 1-naphthaleneacetic acid (NAA). The highest increase in fresh weight (FW) of proliferated SEs (183.33 ± 28.93 mg per 100 mg initial FW) was gained on hormone-free MVW. These SEs eventually developed into vigorous plantlets after culturing on MVW supplemented with 0.2% (w/v) activated charcoal and 50 g L−1 banana homogenate for 12 wk. A histological study revealed that SEs originated from both the epidermal and the sub-epidermal layers of the original protocorm via direct somatic embryogenesis. The genetic homogeneity between the mother generation (V1; the original protocorm) and subsequent generations (V2 and V3; the primary and secondary somatic embryos, respectively) was shown to be identical via random amplification of polymorphic DNA (RAPD) assessment. All regenerated plants grew well under greenhouse conditions.

Micropropagation of Nelumbo nucifera 'Weishan Hong' through germfree mature embryos Abstract Asian lotus (Nelumbo nucifera Gaertn.) is an economically important aquatic plant that is widely cultivated in Asia. It is mainly propagated by dividing rhizomes, which maintains genotype stability but is also laborious and time-consuming. An efficient in vitro method that ensures genetic stability is highly desirable. An optimized method was developed to produce intact and sterile mature embryos by soaking fruits in sterile water containing 2% (v/v) plant preservative mixture. Using mature embryos as explants, an improved micropropagation procedure was established in N. nucifera 'Weishan Hong.' The growth patterns of explants differed with the types and concentrations of cytokinins. Media containing 0.5 to 2.0 mg L−1 kinetin were suitable for elongation of unexpanded stems, while media with 1.0 to 2.0 mg L−1 6-benzylaminopurine (BAP) the stems were shorter. Higher concentrations of about 3.0 mg L−1 BAP promoted shoot cluster formation. Liquid–solid, double-layer media promoted plantlet growth. The well-rooted plantlets could normally grow after being transplanted. Lotus is known to be recalcitrant to regeneration from callus. Apices of mature embryos, aseptic plantlets, and young embryos formed callus successfully when the medium was supplemented with 2,4-dichlorophenoxyacetic acid and thidiazuron. Callus morphology differed with respect to the source. However, only a few of the calluses developed axillary shoots.

Increased freezing stress tolerance of Nicotiana tabacum L. cv. Bright Yellow-2 cell cultures with the medium addition of Ascophyllum nodosum (L.) Le Jolis extract Abstract The study of bioactive components of the brown alga Ascophyllum nodosum (L.) Le Jolis has shown that they promote growth and increase productivity of plants. However, a standardized model system providing consistent responses to such bioactive components has yet to be established. Given that freezing stress, especially in northern climates, reduces plant growth and productivity, a technique was developed to protect plant cells under freezing stress using the natural products made from the abundant A. nodosum as a cell culture supplement. In this study, a homogenous cell culture system of Nicotiana tabacum L. cultivar Bright Yellow-2 (BY-2) suspension cells was used to investigate the bioactivity and protection level of this alga extract under freezing temperatures, and BY-2 cell growth, physiology, and molecular changes were measured in the presence or absence of the extract. The results indicated that A. nodosum extract significantly improved BY-2 cell survival after exposure to freezing temperatures. Inclusion of alga extract also improved cell growth, membrane stability, and nuclear integrity, and reduced cell death of cold-stressed BY-2 suspension cultures. It was concluded that A. nodosum extract influenced cellular and molecular regulation and triggered mechanisms, such as osmolyte accumulation and antioxidant activity, to combat freezing stress in BY-2 suspension cells.

Goji berry ( Lycium barbarum L.) in vitro multiplication improved by light-emitting diodes (LEDs) and 6-benzylaminopurine Abstract In Brazil, Lycium barbarum L. (goji berry) is an agronomically valuable, imported, commercialized plant. The present study evaluated the effects of cytokinins and light sources on optimization of in vitro multiplication of goji berry by axillary bud proliferation. Nodal explants were excised from plants 90 d after in vitro germination and cultured on Murashige and Skoog (MS) medium containing 30 g L−1 sucrose; 7 g L−1 agar-agar; and 0, 2, 4, 8, or 16 μM 6-benzylaminopurine (BAP), 6-furfurylaminopurine (KIN), or thidiazuron (TDZ). After 60 days, regenerated lateral buds and shoot lengths were evaluated. The regenerated shoots were inoculated in MS supplemented with 0.0, 2.5, 5.0, 10.0, or 20.0 μM BAP under white fluorescent lamps or mixed treatment of red/blue (RB) light-emitting diode (LED) lamps. After 60 days in vitro cultivation, the number of formed shoots, leaf number, and shoot length were determined. Subsequently, the shoots were separated and acclimatized in a greenhouse for 30 d and the percentage of survival was evaluated. High regeneration and shoot numbers during lateral bud regeneration occurred with MS medium plus 4 μM BAP. The estimated optimum concentration of 11.56 μM BAP under RB LED lamp resulted in a high shoot number (8.15), with more average leaves per plant (9.2) at 5 μM BAP, and longer shoots (12.34 cm) at 12.42 μM BAP. Acclimatization was successful with 100% survival of ex vitro plants. An efficient in vitro multiplication protocol of an economically important species, goji berry, was developed to facilitate large-scale commercialization of this species.

Wounding and medium formulation affect de novo shoot organogenic responses in hypocotyl-derived explants of annatto ( Bixa orellana L.) Abstract Bixa orellana L. (annatto) is the unique exploited source for the production of bixin and norbixin natural dyes, which are widely used in the food, pharmaceutical, cosmetic, and textile industries. In tissue culture, adequate culture conditions are essential for the development of the plant. Among them, the saline composition of the culture medium and the type of explants are limiting factors. In this study, the effect of MS (Murashige and Skoog), Correia and colleagues (JADS) medium, Woody Plant Medium (WPM), and Driver Kuniyuki Walnut (DKW) culture media and wounding of hypocotyl explants on de novo shoot organogenesis (DNSO) of annatto were evaluated. It was demonstrated that JADS medium improved the nutritional status of shoots, which promoted the best development and quality of the plants generated after acclimatization, when compared to MS medium. The use of JADS medium for annatto DNSO during 45 d of culture was sufficient to regenerate and stimulate the growth of shoots, which had adequate elongation and rhizogenic responses. In addition, enlargement of the wound surface, using longitudinal sections of the hypocotyl explant cut surface down on JADS medium, significantly improved the frequency and number of newly formed shoots. This protocol enabled production of an average of 17 30-mm-long shoots per explant after 90 d of culture. These results will be useful for future studies related to annatto biotechnology.

Evaluation of canavanine as an allelochemical in etiolated seedlings of Vicia villosa Roth: protoplast co-culture method with digital image analysis Abstract Strong allelopathic activities of etiolated seedlings of a leguminous plant, Vicia villosa Roth (hairy vetch), were confirmed by a protoplast co-culture method. The combination of 1% Cellulase RS and 1% Driselase 20 in 0.4 M mannitol solution was used for protoplast isolation. The inhibitory effects of V. villosa protoplasts on the growth of recipient Lactuca sativa L. (lettuce) protoplasts were determined using 96-well culture plates with 50 μL of liquid medium consisting of Murashige and Skoog's basal medium containing 1 μM 2,4-dichlorophenoxyacetic acid, 0.1 μM 6-benzyladenine, 3% (w/v) sucrose, and 0.4 M mannitol in each well. The allelopathic activity of V. villosa epicotyl protoplasts (2 × 102 to 105 mL−1), on cell division of lettuce protoplasts (6 × 103 to 5 × 104 mL−1), was stronger than that of roots and was stronger than on cell wall formation. Digital image analysis of co-cultured protoplasts (DIA-PP method) showed the effect of V. villosa protoplasts on the accumulation of a yellow color in lettuce protoplasts. Weaker inhibition was seen than at the cell wall formation and cell division stages. Effects of putative allelochemical, canavanine at 10 μM strongly inhibited division of lettuce protoplasts, whereas putative allelochemical, cyanamide was stimulatory at 10 μM, but inhibitory at 1 mM. A high level of canavanine was found in epicotyl protoplasts using gas chromatography-mass spectrometry. Canavanine was suggested to be the allelochemical in V. villosa epicotyl protoplasts of etiolated seedlings, although its effect differed with the growth stages of lettuce protoplasts.

A new temporary immersion system for commercial micropropagation of banana ( Musa AAA cv. Grand Naine) Abstract Temporary immersion systems (TIS) are among the best options for commercial micropropagation. However, although successes have been reported with TIS, it is necessary to establish the most suitable system. The aim of this research was to compare the efficiency of different banana (Musa AAA cv. Grand Naine) micropropagation systems. In addition, a liquid medium with partial immersion and a semi-solid culture medium were evaluated. At 28 d of culture, the number and length of shoots, number of leaves per shoot, chlorophyll content, stomatal index, and the percentage of closed stomata were recorded. Additionally, the survival percentage and ex vitro development of the plantlets were evaluated during acclimatization. Results showed that TIS have a higher multiplication rate with respect to partial immersion and semi-solid medium. The chlorophyll content, stomatal index, and stomata functioning were affected by the different culture systems. The different culture systems had no effect on the survival percentage, although morphological differences were observed during acclimatization. The SETIS™ bioreactor was more efficient and resulted in better development in the plants obtained in vitro and ex vitro, which proved this system to be a useful alternative for banana micropropagation.

Bixa orellana L. (achiote) tissue culture: a review Abstract Bixa orellana L. (achiote) is a commercially important plant grown for its natural dye annatto, which is derived from the arils of seeds. Annatto, which contains the antioxidant bixin, is used in food, cosmetic, and textile industries as a natural colorant. Even though B. orellana can be propagated by seed, established tissue culture and in vitro propagation protocols exist for this plant. Organogeneses, both axillary and adventitious, as well as somatic embryogenesis, have been successfully induced from different explant sources, including cotyledons, hypocotyls, roots, stems, and leaves, and effective acclimatization protocols exist for regenerated plantlets. This present mini-review highlights the achievements in the tissue culture of achiote. In addition to using in vitro techniques for the clonal propagation of elite plants, knowledge from these protocols could be used to establish mass production systems, such as bioreactors, to facilitate year-round bixin harvest.

In vitro storage of micropropagated grapevine rootstocks at low temperature Abstract A long-term in vitro conservation protocol for five micropropagated grapevine rootstocks (3309 Couderc, 110 Richter, Vitis riparia Gloire de Montpellier, 101-14 Millardet et De Grasset, and Teleki 5C) was developed and optimized by testing the effect of culture containers (polyethylene bags versus glass test tubes), explant types (rooted versus unrooted cuttings), low-temperature regimes (1.6, 4.0, and 7.2°C), and light intensity (low versusdark). Optimal storage conditions were achieved at 4.0°C and low light intensity (3 μmol m−2 s−1), with rooted plantlets established on woody plant medium (WPM) supplemented with 15 g L−1 sucrose and 37 mg L−1cysteine in polyethylene bags. After 12 and 24 mo of optimal cold storage, the plant condition ratings (3.5 and 2.5 on a scale of 4) and survival rates (75% and 52%) were high, regardless of the rootstock genotype, as determined by visual observations and viability assays of subcultures on WPM supplemented with 15 g L−1 sucrose and 37 mg L−1 cysteine at 25°C, high light intensity (33 to 45 μmol m−2 s−1), and 16-h photoperiod. Prolonged storage at 4°C under low light intensity reduced the number of subcultures by 4- to 8-fold over 12 to 24 mo, which substantially lowered costs and labor, and provided opportunities for long-term germplasm conservation. Additionally, the use of polyethylene bags reduced storage space in the growth room and in a microprocessor temperature-controlled refrigerator, and also facilitated the safe exchange of germplasm across regulatory boundaries.

The in vitro propagation system of Citrus × latifolia (Yu. Tanaka) Yu. Tanaka (Rutaceae) affects the growth and depletion of nutriments Abstract Among citrus fruits, Citrus × latifolia (Yu. Tanaka) Yu. Tanaka stands out given its diverse uses, and there is a great demand for plants of high phytosanitary and genetic quality. In the present study, the in vitromicropropagation process of C. latifolia was studied in RITA®-type bioreactor, semisolid medium, and partial immersion with liquid culture medium. Explants that were 2 mo of age were used and apices of 0.5 cm were extracted and placed in each culture system. In all the systems, MS (Murashige and Skoog) culture medium was used, supplemented with 6-benzylaminopurine (BAP) and kinetin (KIN). After 30 d, the number of shoots, the size of the shoots, the number of leaves, and the depletion of nutrients from the culture medium were analyzed. With the data obtained, ANOVA and Tukey's mean comparison tests were performed. The RITA® bioreactors, with the addition of 40 mL of medium per explant in a 4-h-interval immersion, facilitated the production of the greatest number of shoots compared to the other culture systems, and also resulted in the highest levels in medium nutrient depletion.

Radiology

Forensic Radiology: An Exciting and Developing Field That Needs More Trained Radiologists Publication date: Available online 27 April 2019 Source: Academic Radiology Author(s): Barry Daly

Paired Drainage Catheter Insertion: Feasibility of Placing Two Catheters within the Same Complex Abscess Cavity as a Primary and Salvage Percutaneous Drainage Technique Publication date: Available online 26 April 2019 Source: Academic Radiology Author(s): David H. Ballard, Sarah T. Flanagan, Ryan W. Brown, Romulo Vea, Chaitanya Ahuja, Horacio B. D'Agostino Rationale and Objectives To assess the feasibility of paired catheter drainage for management of complex abdominal abscesses. Materials and Methods This was a single-center retrospective study of 54 patients (35 males; mean age 48.9 years) that underwent paired catheter insertion for complex abdominal fluid collections in an 18-month period. Complex collections were defined as abscesses ≥6 cm in diameter with septations, high viscosity fluid or necrotic debris, or abscesses with an associated fistula. Abscess etiologies included postoperative (n = 28), pancreatitis (n = 12), perforated bowel (n = 7), liver abscess (n = 4), and perihepatic from gallbladder perforation (n = 3). Paired catheter insertion was defined two catheters co-located within one collection through the same skin incision or two closely spaced insertion sites. Paired catheter insertion was used primarily as initial drainage for complex intraabdominal abscesses and for salvage drainage in collections that could not be evacuated by a single catheter. Primary paired catheter insertion was used in 45 patients and as salvage in nine patients. Results Abscess resolution occurred in 51 (94.4%) patients. Patients had a median of three drainage procedures. Median duration of paired catheterization was 22 days. Seven abscesses recurred and all resolved with repeated drainage. Complications included one hemorrhage that was taken for surgical exploration. Overall, 48 patients had good clinical outcome, 3 patients died (multiorgan failure, n = 2; sepsis, n = 1), and 3 patients were lost to follow-up. Conclusion Percutaneous paired catheter drainage is a feasible technique for the treatment of both complex intraabdominal abscesses and abscesses unresponsive to single catheter drainage.

Liver Ultrasound Texture Analysis: The Computer Finds More to Quantify Than Meets the Eye Publication date: Available online 25 April 2019 Source: Academic Radiology Author(s): Andrew Phelps

Reducing Radiation Dose and Improving Image Quality in CT Portal Venography Using 80 kV and Adaptive Statistical Iterative Reconstruction-V in Slender Patients Publication date: Available online 25 April 2019 Source: Academic Radiology Author(s): Zhanli Ren, Xirong Zhang, Zhijun Hu, Dou Li, Zhentang Liu, Donghong Wei, Yongjun Jia, Nan Yu, Yong Yu, Yuxin Lei, Xiaoxia Chen, Changyi Guo, Zhanliang Ren, Taiping He Objective To explore the feasibility of reducing radiation dose and improving image quality in CT portal venography (CTPV) using 80 kV and adaptive statistical iterative reconstruction-V(ASIR-V) in slender patients in comparison with conventional protocol using 120 kV and ASIR. Methods Sixty slender patients for enhanced abdominal CT scanning were randomly divided into group A and group B. Group A used the conventional 120 kV tube voltage, 600 mgI/kg contrast dose and reconstructed with the recommended 40% ASIR. Group B used 80 kV tube voltage, 350 mgI/kg contrast dose and reconstructed with ASIR-V from 40% to 100% with 10% interval. The CT values and standard deviation (SD) values of the main portal vein, left branch, and right branch of portal vein, liver, and erector spinae at the same level were measured to calculate the signal-to-noise ratio (SNR) and contrast-to-noise ratio (CNR). The image quality was subjectively scored by two experienced radiologists blindly using a 5-point criterion. The contrast dose, volumetric CT dose index, and dose length product were recorded in both groups and the effective dose was calculated. Results There was no significant difference in general data between the two groups (p > 0.05), the effective dose and contrast dose in group B were reduced by 63.3% (p < 0.001) and 39.7% (p < 0.001), respectively compared with group A. With the percentage of ASIR-V increased in group B, the CT values showed no significant difference, while the SD values gradually decreased and SNR values and CNR values increased accordingly. Compared with group A, group B demonstrated similar CT values (p > 0.05), while the SD values with 80% ASIR-V to 100% ASIR-V were significantly lower than those of 40% ASIR (p < 0.001), and the SNR values and CNR values with 70% ASIR-V to 100% ASIR-V were significantly higher than those of 40% ASIR (p < 0.001). The subjective image quality scores by the two radiologists had excellent consistency (kappa value>0.75, p < 0.001), and the final subjective image quality scores and the subjective scores in each of the 5 scoring categories with 60% ASIR-V to 100% ASIR-V were all significantly higher than those of 40% ASIR, and 80% ASIR-V obtained the highest subjective score among different reconstructions. Conclusion In CTPV, the application of 80 kV and ASIR-V reconstruction in slender patients can significantly reduce radiation dose (by 63.3%) and contrast agent dose (by 39.7%). Compared with the recommended 40% ASIR using 120 kV, ASIR-V with 80% to 100% percentages can further improve image quality and with 80% ASIR-V being the best reconstruction algorithm. Advances in knowledge CTPV with 80 kV and ASIR-V algorithm in slender patients can significantly reduce radiation dose and contrast agent dose as well as improve image quality, compared with the conventional 120 kV protocol using 40% ASIR.

An Appreciative Approach to Burnout Publication date: Available online 18 April 2019 Source: Academic Radiology Author(s): Richard B Gunderman

Making Learning Fun: Gaming in Radiology Education Publication date: Available online 17 April 2019 Source: Academic Radiology Author(s): Omer Awan, Courtney Dey, Hayden Salts, James Brian, Joseph Fotos, Eric Royston, Maria Braileanu, Emily Ghobadi, Jason Powell, Charlotte Chung, William Auffermann With continued technologic advances, it is not surprising that gaming techniques are increasingly being used in radiology residency programs. This comprehensive review on gaming in radiology education offers insight into the importance of gaming, types of games and principles utilized in gaming, as well as applications that are inherent in artificial intelligence and continued medical education. The advantages and disadvantages of gaming will be considered, as well as barriers to successful adoption of gaming.

Forensic Radiology: A Primer Publication date: Available online 17 April 2019 Source: Academic Radiology Author(s): Summer J. Decker, Maria Braileanu, Courtney Dey, Leon Lenchik, Michael Pickup, Jason Powell, Maria Tucker, Linda Probyn Rationale and objective: Forensic radiology is a relatively unknown subspecialty which is becoming increasingly more important. The field incorporates antemortem and postmortem imaging for the detection and documentation of various pathologies for medicolegal purposes. Postmortem imaging is increasingly used in conjunction with the traditional autopsy in a process called a "virtual" autopsy. Radiography has been a staple of forensic investigations for over a century, first used in 1896. Advanced imaging techniques such as postmortem computed tomography and postmortem magnetic resonance imaging have only recently gained acceptance in the forensic science community. Postmortem computed tomography and postmortem magnetic resonance imaging methods are now widely used in some parts of the world, while other countries including the United States have been slower to adopt these methods into their daily practice. Advanced forensic imaging is increasingly used in the courts where juries have responded positively to such presentation of forensic data. For these reasons, advanced postmortem imaging is becoming a regular part of forensic investigations. The increase in the use of forensic imaging presents a unique opportunity for radiologists to collaborate with pathologists and law enforcement officials. This paper provides an overview of forensic radiology and identifies potential challenges and opportunities.

Discrepancy in Practices Related to the Use of Oral Contrast in Abdominal CT Scan Publication date: Available online 13 April 2019 Source: Academic Radiology Author(s): Shahmeer Khan, Muhammad Awais, Anwar Ahmed

Complicated Cysts: Should Management Depend Upon Age? Publication date: Available online 13 April 2019 Source: Academic Radiology Author(s): Dana Ataya, Bethany L. Niell

Impact of Patient Size and Radiation Dose on Accuracy and Precision of Iodine Quantification and Virtual Noncontrast Values in Dual-layer Detector CT—A Phantom Study Publication date: Available online 12 April 2019 Source: Academic Radiology Author(s): Steven Van Hedent, Curtis Tatsuoka, Sarah Carr, Kai Roman Laukamp, Brendan Eck, Nils Große Hokamp, Rivka Kessner, Pablo Ros, David Jordan Rationale and Objectives Iodine quantification (IQ) and virtual noncontrast (VNC) images produced by dual-energy CT (DECT) can be used for various clinical applications. We investigate the performance of dual-layer DECT (DLDECT) in different phantom sizes and varying radiation doses and tube voltages, including a low-dose pediatric setting. Materials and Methods Three phantom sizes (simulating a 10-year-old child, an average, and a large-sized adult) were scanned with iodine solution inserts with concentrations ranging 0-32 mg/ml, using the DLDECT. Each phantom size was scanned with CTDIvol 2-15 mGy at 120 and 140 kVp. The smallest phantom underwent additional scans with CTDIvol 0.9-1.8 mGy. All scans were repeated 3 times. Each iodine insert was analyzed using VNC and IQ images for accuracy and precision, by comparison to known values. Results For scans from 2 to 15 mGy mean VNC attenuation and IQ error in the iodine inserts in the small, medium, and large phantoms was 1.2 HU ± 3.2, −1.2 HU ± 14.9, 2.6 HU ± 23.6; and +0.1 mg/cc ± 0.4, −0.9 mg/cc ± 0.9, and −1.8 mg/cc ± 1.8, respectively. In this dose range, there were no significant differences (p ≥ 0.05) in mean VNC attenuation or IQ accuracy in each phantom size, while IQ was significantly less precise in the small phantom at 2 mGy and 10 mGy (p < 0.05). Scans with CTDIvol 0.9-1.8 mGy in the small phantom showed a limited, but statistically significantly lower VNC attenuation precision and IQ accuracy (−0.5 HU ± 5.3 and −0.3 mg/cc ± 0.5, respectively) compared to higher dose scans in the same phantom size. Conclusion Performance of iodine quantification and subtraction by VNC images in DLDECT is largely dose independent, with the primary factor being patient size. Low-dose pediatric scan protocols have a significant, but limited impact on IQ and VNC attenuation values.

Addictive Behaviors

Associations among simultaneous and co-occurring use of alcohol and marijuana, risky driving, and perceived risk Publication date: September 2019 Source: Addictive Behaviors, Volume 96 Author(s): Jennifer C. Duckworth, Christine M. Lee

Shifts in momentary motivation to quit smoking based on experimental context and perceptions of motivational instability Publication date: September 2019 Source: Addictive Behaviors, Volume 96 Author(s): Jennifer C. Veilleux Abstract Background Smoking quit intentions (i.e., plans or commitments to reduce smoking) are distinct from quit motivation (i.e., desire to quit). Motivation to quit is a dynamic avoidance motivational state, likely subject to frequent fluctuations and shifts based on environmental context, similar to craving (an approach motivational state). Moreover, people's perceptions of motivational instability may influence their momentary quit motivation and beliefs surrounding their abilities to manage the quit process successfully. Methods In the current study, smokers were recruited online. Baseline motivation to quit was examined along with perceptions of motivational instability. Participants who reported some interest in eventually quitting (n = 174; Mage = 45.29, SD = 14.81, 74.1% women) were randomly assigned to negative affect, cigarette or neutral cue conditions and then momentary motivation to quit was assessed, followed by a validated index of quit intentions. Results Participants in the negative affect and cigarette cue conditions reported greater momentary motivation to quit than participants in the neutral condition (F = 3.73, p = .03), particularly for people who reported upfront that their motivation did not change from day-to-day (F = 3.97, p = .02). In addition, greater momentary quit motivation predicted stronger quit intentions (B = 0.66, p < .001). Conclusion Findings from this study suggest that motivation to quit is likely dynamic and influenced by context as well as by self-perceptions. Implications suggest that further efforts to separate motivation from intention may be useful in understanding smokers' desires and plans to quit smoking.

Evaluating the predictive value of measures of susceptibility to tobacco and alternative tobacco products Publication date: September 2019 Source: Addictive Behaviors, Volume 96 Author(s): Jessica L. Barrington-Trimis, Fei Liu, Jennifer B. Unger, Todd Alonzo, Tess Boley Cruz, Robert Urman, Mary Ann Pentz, Kiros Berhane, Rob McConnell Abstract Background The "cigarette susceptibility index" has been adapted for other products, yet, the validity of these adapted measures–particularly among youth who have used other tobacco products–has not been evaluated. Methods We used prospective data from the Southern California Children's Health Study to evaluate the association of questionnaire measures assessing susceptibility to e-cigarette, cigarette, hookah and cigar/cigarillo/little cigar use at wave 1 (W1; 11th/12th grade) with subsequent initiation between W1 and W2 (16 months later, N = 1453). We additionally examined whether each effect estimate differed by use of other tobacco products at W1. Results Odds ratios, attributable risk%, and risk differences for product initiation among susceptible vs. non-susceptible youth were consistently higher among never users of any tobacco product than among youth with any tobacco use history. For example, susceptible (vs. non-susceptible) youth with no prior tobacco use had 3.64 times the odds of subsequent initiation of e-cigarettes (95%CI:2.61,5.09), while among users of another product, susceptible (vs. non-susceptible) youth had 1.95 times the odds of e-cigarette initiation (95%CI:0.98,3.89; p-interaction = 0.016). 60.4% of e-cigarette initiation among never users of any product could be attributed to susceptibility, compared to 19.8% among users of another product. The e-cigarette absolute risk difference between susceptible and non-susceptible youth was 21.9%(15.2,28.6) for never users, vs. 15.4%(0.2,30.7) for users of another product. Conclusion Tobacco product-specific susceptibility associations with initiation of use at W2 were markedly attenuated among prior users of other products, demonstrating reduced utility for these measures among subjects using other products.

Peer victimization and problematic internet use in adolescents: The mediating role of deviant peer affiliation and the moderating role of family functioning Publication date: September 2019 Source: Addictive Behaviors, Volume 96 Author(s): Boyu Zhai, Dongping Li, Jichao Jia, Yuxiao Liu, Wenqiang Sun, Yanhui Wang Abstract The role of social-environmental factors in adolescent problematic Internet use (PIU) has attracted considerable attention recently. Several studies have documented that peer victimization is positively associated with PIU. However, little is known about "how" (i.e., mediation mechanisms) and "under what conditions" (i.e., moderation mechanisms) peer victimization is associated with adolescent PIU. To contribute to this gap in the knowledge, this study used a large sample of Chinese adolescents (N = 2758; Mage = 13.53 years, SD = 1.06) to examine deviant peer affiliation (DPA) as a mediator and family functioning as a moderator in this relationship. Students completed anonymous questionnaires to measure the main variables. After controlling for important covariates related to PIU, the results indicated that (a) peer victimization was positively associated with PIU, (b) DPA partially mediated the link between peer victimization and PIU, and (c) family functioning moderated the association between peer victimization and DPA. Specifically, for adolescents with better family functioning, the relationship between peer victimization and DPA was weaker. The current research deepens our understanding of "how" and "under what conditions" peer victimization is related to PIU in adolescents.

Associations between loot box use, problematic gaming and gambling, and gambling-related cognitions Publication date: September 2019 Source: Addictive Behaviors, Volume 96 Author(s): Gabriel A. Brooks, Luke Clark Abstract Loot boxes are virtual goods in video games that produce randomly-generated in-game rewards, and have attracted scrutiny because of a resemblance to gambling. This study tests relationships between gaming involvement, engagement with loot boxes, and their associations with disordered gambling and gambling-related cognitions. Online questionnaires were completed by 144 adults via MTurk (Study 1) and 113 undergraduates (Study 2). Gaming and loot box-related variables included estimated time spent gaming and monthly expenditure, the Internet Gaming Disorder Scale (IGDS), and questions that assessed perceptions and behaviours related to loot boxes. Most participants thought loot boxes were a form of gambling (68.1% & 86.2%). A subset of items were condensed into a unidimensional "Risky Loot-box Index" (RLI) via exploratory factor analysis. In Study 1, the RLI showed significant associations with the Problem Gambling Severity Index (r = .491, p < .001) and the Gambling Related Cognitions Scale (r = .518, p < .001). Overall, gambling-related variables predicted 37.1% (p < .001) of the variance in RLI scores. Findings were replicated, though attenuated, in Study 2. These results demonstrate that besides the surface similarity of loot boxes to gambling, loot box engagement is correlated with gambling beliefs and problematic gambling behaviour in adult gamers.

Heavy episodic drinking in late adolescents: The role of theory of mind and conformity drinking motives Publication date: September 2019 Source: Addictive Behaviors, Volume 96 Author(s): Fiorenzo Laghi, Dora Bianchi, Sara Pompili, Antonia Lonigro, Roberto Baiocco Abstract Background Heavy episodic drinking (HED) implies severe consequences for health, both in the short and long term. Peer conformity has shown contradictory relationships with HED, suggesting the presence of a complex interaction between social and individual factors. Objective This study investigated the moderation role of conformity in the relationship between theory of mind (ToM) and HED. Method Participants were 170 Italian adolescents (Mage = 18.71, SDage = 0.77) recruited in public high schools. Self-report measures were administered for evaluating drunkenness and HED. The Drinking Motives Questionnaire was used to assess three drinking motives, namely coping with negative emotions, enhancement of positive feelings, and the need for peer conformity, whereas the Reading the Mind in the Eyes Test was adopted to test advanced theory of mind skills. ANOVAs, MANOVAs and regression moderation analyses were performed. Results Drunkenness, HED and drinking motives did not vary with gender. Only social motives were higher in boys (vs. girls). HED was positively related to drunkenness and to social, coping and enhancement motives. However, only enhancement significantly predicted HED. Finally, conformity moderated the relationship between ToM and HED. In the presence of high conformity, adolescents with impaired ToM reported significantly more HED than adolescents with high ToM. Conclusions In the presence of conformity needs, social cognition impairment appeared to be a risk factor for HED, whereas good socio-cognitive skills were protective. Implications for research, clinical assessment and prevention are discussed.

Cross-cultural examination of college marijuana culture in five countries: Measurement invariance of the Perceived Importance of Marijuana to the College Experience Scale Publication date: September 2019 Source: Addictive Behaviors, Volume 96 Author(s): Matthew R. Pearson, Adrian J. Bravo, Melissa Sotelo, Cross-Cultural Addictions Study Team Abstract Marijuana internalized norms, measured by the Perceived Importance of Marijuana to the College Experience Scale (PIMCES; 8 items), has been found to be distinct from marijuana descriptive/injunctive norms and to be a unique robust predictor of marijuana-related outcomes among college students, yet the role of these beliefs has not been studied outside the U.S. Using confirmatory factor analysis, the present work examined the level of measurement invariance (i.e., configural, metric, and scalar) of the PIMCES across five different countries with distinct marijuana-related regulations (i.e., U.S., Argentina, Spain, Uruguay, and the Netherlands), sex, and marijuana user status among college students (n = 3424) recruited between September 2017 and January 2018. To make valid comparisons across groups, metric invariance is needed to compare correlations and scalar invariance is needed to compare latent means. We found strong measurement invariance (i.e., scalar invariance) for the PIMCES across countries, across males and females, and across marijuana users and non-users. College students in the U.S. reported the highest levels of marijuana internalized norms compared to college students from all other countries. As expected, males and marijuana users showed significantly higher scores on the PIMCES than females and non-users, respectively. Bivariate correlations between PIMCES scores and other marijuana-related variables were remarkably similar across males and females, though differences across countries warrant further exploration. Taken together, the degree to which college students view marijuana use to be an integral part of the college experience may be an important target for college student marijuana interventions across various countries/cultures.

Psychiatric comorbidity among first-time and repeat DUI offenders Publication date: September 2019 Source: Addictive Behaviors, Volume 96 Author(s): Layne M. Keating, Sarah E. Nelson, Rhiannon C. Wiley, Howard J. Shaffer Abstract Driving under the influence of alcohol or other substances is a serious public health concern. Previous research has shown that psychiatric comorbidity is more prevalent for repeat offenders than the general population, and that first-time offenders exhibit elevated rates of psychiatric comorbidity, but few studies have directly compared first-time and repeat DUI offenders. The current study compares psychiatric comorbidity among repeat and first-time DUI offenders. First-time and repeat DUI offenders completed the screener module of the Computerized Assessment and Referral System (CARS), adapted from the Composite International Diagnostic Interview (CIDI: Kessler & Ustun, 2004), to measure potential psychiatric comorbidity. For 16 of 19 psychiatric disorders, repeat DUI offenders were more likely to screen positive during their lifetime compared with first-time DUI offenders. Similarly, repeat DUI offenders were more likely to screen positive during the past year for 11 of 16 assessed psychiatric disorders. Overall, repeat DUI offenders screened positive for an average of 6.3 disorders during their lifetime, compared to first-time offenders who screened positive for an average of 3.7 disorders. Repeat DUI offenders also screened positive for more past-year disorders (M = 3.3) than first-time offenders (M = 1.9). Compared to first-time offenders, repeat DUI offenders evidence more severe and pervasive psychiatric comorbidity. Further research is necessary to determine whether psychiatric comorbidity among first-time offenders directly predicts re-offense. If so, screening for mental health issues among first-offenders could provide valuable information about how best to allocate resources for these offenders.

Cocaine use is declining among emerging adults in the United States: Trends by college enrollment Publication date: September 2019 Source: Addictive Behaviors, Volume 96 Author(s): Kristin E. Schneider, Julie K. Johnson, Renee M. Johnson Abstract Background The difference in cocaine use between student and non-student emerging adults is not well understood, despite the longstanding assumption that college is protective against use. Objective To describe trends in cocaine use among emerging adults by college enrollment. Methods Using cross-sectional, nationally-representative data from the National Survey on Drug Use and Health (2002–2016), we estimated the prevalence of four cocaine use indicators by year among emerging adults (ages 18–22) and stratified by college enrollment. We tested for linear and quadratic trends over time. Results From 2002 to 2016, 11.7% reported lifetime cocaine use, 5.8% reported past 12-month use, 1.7% reported past 30-day use, and 1% had a use disorder in the past 12-months. We observed significant linear decreases in all four use indicators over time (Lifetime: β = −0.20, Past 12-months: β = −0.17, Past 30-days: β = −0.22, Past 12-months disorder: β = −0.37). Students had lower rates of cocaine use than non-students, although prevalence estimates were more similar between groups for past 12-month (Students:5.3%, 95% CI:5.0–5.5; Non-students:6.3%, 95% CI:6.1–6.5) and past 30-day use (Students:1.6%, 95% CI:1.4–1.7; Non-students:1.8%, 95% CI:1.7–1.9) than for lifetime use (Students:9.4%, 95% CI:9.1–9.7; Non-students:13.8%, 95% CI:13.5–14.1). Conclusions Cocaine use has declined among emerging adults since the early 2000s. The college environment appears compensate for early life differences in cocaine use risk between students and non-students, closing the gap between groups for recent indicators of cocaine use. Given the severe health consequences associated with cocaine use, more public health attention to this issue is needed.

Desire thinking as a predictor of drinking status following treatment for alcohol use disorder: A prospective study Publication date: August 2019 Source: Addictive Behaviors, Volume 95 Author(s): Francesca Martino, Gabriele Caselli, Elena Fiabane, Federica Felicetti, Cecilia Trevisani, Marco Menchetti, Clarice Mezzaluna, Sandra Sassaroli, Ian P. Albery, Marcantonio M. Spada Abstract Research has indicated that craving is one of the strongest predictors of treatment outcome and relapse in Alcohol Use Disorders (AUD) but there is little consensus on the factors that may influence its activation and escalation. Research has also shown that desire thinking is an important cognitive process which may exacerbate craving in problem drinkers. The aim of present study was to explore, for the first time, the role of desire thinking in prospectively predicting relapse, craving and binge drinking in patients receiving treatment for AUD. One hundred and thirty-five patients admitted to two rehabilitation centres and two outpatient services for addiction and mental health problems were administered baseline, treatment completion and three months follow-up measures of anxiety and depression, AUD severity, binge drinking frequency, craving and desire thinking. Results indicated that the verbal perseveration component of desire thinking at treatment completion was the only significant predictor of relapse at follow-up over and above baseline AUD severity and binge drinking frequency. Furthermore, the imaginal prefiguration component of desire thinking and craving levels at treatment completion were found to predict craving levels at follow-up independently of AUD severity and binge drinking frequency at baseline. Finally, both the imaginal prefiguration and verbal perseveration components of desire thinking at treatment completion were found to be the only predictors of binge drinking frequency at follow-up independently of AUD severity and binge drinking frequency at baseline. Treatments for AUD should aim to reduce desire thinking in people to enhance clinical outcomes and reduce relapse risk.

Palaeogeography, Palaeoclimatology, Palaeoecology

3. Easier access to "fossil gelatin" and quality check by amino acid racemization Publication date: 15 July 2019 Source: Palaeogeography, Palaeoclimatology, Palaeoecology, Volume 526 Author(s): Hartwig Elster Abstract Gelatin in fossil bones is the most important source for radiocarbon, stable isotopes, ZooMS (Zooarchaeology by Mass Spectrometry) and Amino Acid Racemization (AAR) studies. The isolation and cleaning of fossil gelatin is a time consuming procedure and can introduce contaminations, which, as a consequence, can produce erroneous results. By synthesizing and optimizing a "Molecularly Imprinted Polymer" (MIP) for gelatin this isolation procedure can be faster and more specific. This first application of MIP on fossil gelatin is described. By optimizing the sample preparation over all steps from getting fossil bone powder (ca. 20 mg), isolating the fossil gelatin by MIP, and analyzing the amino acids, especially the enantiomers of aspartic acid from the fossil gelatin by gas chromatography (GC), results are available within one day. These results include a measure of the quality of the fossil gelatin by the pattern of the single amino acids and the ratios of defined amino acids from the GC chromatogram. At the same time, the AAR can give an estimate of the relative age of the fossil bones.

Composition and structural features of two Permian parareptile (Deltavjatia vjatkensis, Kotelnich Site, Russia) bone fragments and their alteration during fossilisation Publication date: 15 July 2019 Source: Palaeogeography, Palaeoclimatology, Palaeoecology, Volume 526 Author(s): Daria Kiseleva, Oleg Shilovsky, Evgeny Shagalov, Anastasia Ryanskaya, Maria Chervyakovskaya, Elizaveta Pankrushina, Nadezhda Cherednichenko Abstract The Kotelnich vertebrate fossil site, one of the richest of the Permian period, is characterised by the excellent preservation of fossil remains due to their burial in a silty anaerobic environment similar to modern peat bogs. The aim of the work is to carry out a comprehensive study of the composition and structure of bone fragments of the Permian parareptile Deltavjatia vjatkensis, as well as that of its embedding rocks, to support further paleoecological and fossilisation reconstructions. Fossil bones and their surrounding strata were investigated using optical and scanning electron microscopy, Raman spectroscopy, X-ray diffraction and inductively coupled plasma mass spectrometry (ICP-MS). The carbonate-rich composition of the sediment was found to favour the preservation of the bone apatite. The mineral phase of the bone tissue corresponds to fluorapatite. Organic matrix vibration bands arising on the Raman spectra correspond to CH vibrations of lipids and proteins and possibly cholesterol. The obtained data indicate a high degree of integrity of both bone mineral phase and its organic proteinaceous residue. White blood cell (WBC)-like structures were detected, which possibly could be interpreted as leukocytes. Elemental mapping of fossil bones was used to reveal element distribution features (uniform or with zones of apparent enrichment-depletion) and secondary mineralisation zones. Ca, P, Mg, Mn and S are uniformly distributed through the bone; conversely, elevated quantities of Fe are assigned to cracks and large pores formed as a consequence of the decomposition of organic materials. As compared to bone apatite, the authigenic calcite filling of bone porosities is significantly depleted in terms of microelements. The apatite rare earth element (REE) composition of bone tissue, which is inherited from seawater, can be used for paleoreconstructions of redox conditions and water composition. La/Sm ratios (0.4–0.5), high Y/Ho ratios, high uranium content and high La/Yb ratios (2–3) in the bone apatite indicate the absence of recrystallisation during the late stages of diagenesis. A positive cerium anomaly detected in the bones indicates the presence of an oxidising and possibly alkaline environment during the early stages of diagenesis in shallow coastal basin of the Kotelnich paleolocality. It is shown that 10 to 20% of REE in bone apatite is derived from seawater absorption (hydrogenous source), with the remainder having been derived from host rock (lithogenic) sources.

Genesis, modification, and preservation of complex Upper Ordovician hardgrounds: Implications for sequence stratigraphy and the Great Ordovician Biodiversification Event Publication date: 15 July 2019 Source: Palaeogeography, Palaeoclimatology, Palaeoecology, Volume 526 Author(s): Timothy R. Paton, Carlton E. Brett, George E. Kampouris Abstract Hardgrounds from the Upper Ordovician strata of eastern Laurentia exhibit a wide range of morphologies, though the development of their complex surface topographies remains poorly understood. These early lithified seafloors, which can be laterally very extensive, may provide key information about basin dynamics and global ocean fluctuations. Based on spectacular exposures and newly-excavated material, we examined a suite of hardgrounds from the Upper Ordovician (Katian) of southern Ontario and the Cincinnati Arch region and documented the processes of genesis, development, and modification of simple, complex, and block hardgrounds. In this study, we used extensive field work, petrology, and cathodoluminescence to determine the evolution and shaping of hardgrounds as a result of intense submarine erosion and prolonged exposure of these surfaces and reveal a progression from simple to complex hardgrounds and ultimately to isolated cobble- to boulder-sized block hardgrounds. We survey the distribution of hardgrounds in a sequence stratigraphic framework to determine allocyclic controls on hardground development, highlight the important role that mixed siliciclastic-carbonate environments play in promoting the development of heterogeneous hardground topographies, and analyze the effects of high surface reliefs on local depositional and erosional regimes. In addition, we interpret the impact of increasingly abundant complex hardgrounds throughout the Ordovician on hard-substrate community diversification and their contribution to the Great Ordovician Biodiversification Event. We also discuss time-specific aspects of the Late Ordovician that made it a peak interval of hardground proliferation.

Chronostratigraphic distribution of cetaceans in the Pliocene of Northern Apennines (Italy): palaeoecological implications Publication date: 15 July 2019 Source: Palaeogeography, Palaeoclimatology, Palaeoecology, Volume 526 Author(s): Alessandro Freschi, Simone Cau, Paola Monegatti, Marco Roveri Abstract A large number of cetacean fossils have been recovered through the time from the Pliocene deposits in Northern Apennines thrust belt, on both the Padan-Adriatic and the Tyrrhenian side. In this work, the cetaceans of Castell'Arquato Basin (Padan-Adriatic side) have been placed within a high-resolution chronostratigraphic framework available for this area. This results in a better assessment of their evolutionary history and of the palaeoecologic factors controlling their diversity and abundance through the time. Our results document a greater diversity between 3.1 and 2.7 Ma time interval, likely due to the development of eutrophic conditions during precessionally-driven insolation maxima at 400 ka eccentricity maxima, which are recorded in deeper depositional settings by sapropel clusters. Eutrophic conditions kept a complex trophic chain, as currently observed in the Ligurian Sea (Corsican-Ligurian-Provençal Basin), an area characterized also by upwelling. A reduction in both number and ecological diversity of cetacean taxa is observed along the Pliocene-Pleistocene transition; this trend, related to the onset of the Quaternary icehouse conditions, appears in good agreement with the recently recognized extinction event that affecting the marine megafauna at the end of the Pliocene.

Distinguishing core and flank facies based on shell fabrics in Lower Jurassic lithiotid shell beds Publication date: 15 July 2019 Source: Palaeogeography, Palaeoclimatology, Palaeoecology, Volume 526 Author(s): Valentina Brandolese, Renato Posenato, James H. Nebelsick, Davide Bassi Abstract Lower Jurassic larger bivalves, mostly represented by the monospecific lithiotid genera Lithiotis, Cochlearites and Lithioperna, formed large shell accumulations in the shallow-water carbonate Tethyan and Panthalassa margins. A quantitative analysis of lithiotid accumulations from the Trento Platform (northern Italy) was carried out in order to distinguish the distribution of autochthonous, parautochthonous and allochthonous lithiotids in the accumulation core and flanks. Various representative accumulations are characterized with respect to systematic content, shell cover, shell density, disarticulation, shell fragmentation and orientation of individual shells. Lithioperna shows a high shell cover within the accumulation core while that of Cochlearites is variable and Lithiotis is rare. All three lithiotids occur in high densities in the accumulation flanks. As expected, autochthonous individuals of all three genera are frequent in the accumulation cores where they are preserved in life position. In the flanks, most bivalves are parautochthonous except for Lithioperna which can grow in life position. All bivalves in allochthonous shell beds are highly fragmented and disarticulated. The studied traits are potentially useful as proxies for distinguishing core or flank facies when the accumulation does not crop out as a whole. Graphical abstract

Carbonate thermoluminescence and its implication for marine productivity change during the Permian-Triassic transition Publication date: 15 July 2019 Source: Palaeogeography, Palaeoclimatology, Palaeoecology, Volume 526 Author(s): Zhipu Qiu, Haijun Song, Chaoyong Hu, Paul B. Wignall, Huyue Song Abstract The Permian-Triassic (P-Tr) crisis was the largest mass extinction of the Phanerozoic and eliminated over 90% of marine species. However, the nature of marine productivity changes during the crisis is a matter of on-going debate. Here, thermoluminescence (TL) measurements from 144 bulk carbonate samples from Meishan (South China) show two levels of variation in TL peak intensities at ~270 °C. The first-order variation is characterized by a rapid, nearly three-fold increase. A secondary variation is interpreted to record periodic fluctuations on a Milankovitch scale (~20-kyr rhythm). The periodic variations of TL are negatively correlated with Mn concentrations, suggesting primary productivity is a key factor controlling the TL peak intensity by photo-reduction of particulate Mn in the presence of organic matter. Therefore, the periodic fluctuations of TL were likely controlled by primary productivity changes on orbital timescales. A significant rise of TL peak intensities across the P-Tr boundary represents the rapid increase of dissolved Mn associated with enhanced marine productivity and/or oceanic anoxia.

Population response during an Oceanic Anoxic Event: The case of Posidonotis (Bivalvia) from the Lower Jurassic of the Neuquén Basin, Argentina Publication date: 1 July 2019 Source: Palaeogeography, Palaeoclimatology, Palaeoecology, Volume 525 Author(s): Sonia Ros-Franch, Javier Echevarría, Susana E. Damborenea, Miguel O. Manceñido, Hugh C. Jenkyns, Aisha Al-Suwaidi, Stephen P. Hesselbo, Alberto C. Riccardi Abstract Benthonic marine species show a wide range of biological reactions to seawater chemical changes through time, from subtle adjustments to extinction. The Early Toarcian Oceanic Anoxic Event (T-OAE) was recently recognized in the Neuquén Basin, Argentina, confirming its global scope. The event was identified chemostratigraphically on the basis of a relative increase in marine organic carbon and a characteristic negative carbon-isotope excursion (δ13Corg) in bulk rock and fossil wood in the upper Pliensbachian–lower Toarcian interval in the Arroyo Lapa section (Neuquén). Simultaneously with collection of lithological samples, a high-resolution biostratigraphical survey was carried out, and the scarce benthonic fauna was collected in order to check the biotic response to changing marine geochemical conditions. We present here an analysis of size and abundance data from the T-OAE interval in the Neuquén Basin for the dominant bivalve species, the paper-clam Posidonotis cancellata (Leanza), and relate these data to geochemical proxies (%TOC and δ13Corg) obtained at the same locality. The abundance of P. cancellata increased when the rest of the benthos diminished, reaching a maximum at the onset level of the T-OAE, and then decreasing. Size-frequency distributions show a noteworthy lack of juvenile shells. Shell size shows a positive correlation with %TOC in the whole section, though over the T-OAE interval proper, it decreases below the level where the maximum %TOC value is attained and increases above it. Posidonotis cancellata shows features of opportunistic species, such as high tolerance to hypoxia, strong dominance in impoverished environments and a strong dependence on primary productivity, but at the same time had a reproductive strategy more similar to equilibrium species, with relatively low juvenile mortality rates. Several anatomical features suggest adaptation to permanently dysaerobic environments. The species disappeared just before the minimum negative carbon-isotope value was reached; and by the same time the genus became extinct worldwide. Graphical abstract

Recognition of tuffs in the middle-upper Dingqinghu Fm., Lunpola Basin, central Tibetan Plateau: Constraints on stratigraphic age and implications for paleoclimate Publication date: 1 July 2019 Source: Palaeogeography, Palaeoclimatology, Palaeoecology, Volume 525 Author(s): Ziqiang Mao, Qingquan Meng, Xiaomin Fang, Tao Zhang, Fuli Wu, Yibo Yang, Weilin Zhang, Jinbo Zan, Mengqi Tan Abstract The Lunpola Basin plays a key role in determining the elevation history of the central Tibetan Plateau, which is closely related to uplift dynamics and paleoclimatic evolution. However, accurate age constraints on the basin stratigraphy reflecting tectonic, paleo-elevation and climate information have long been vigorously debated, mostly due to a lack of absolute age controls. Here, we found two altered tuff layers in the middle-upper Dingqinghu Formation (Fm.) in the basin stratigraphic sequence. Detailed lithological, mineralogical and geochemical analyses of the tuffs indicate that they came from trachyandesite and identify analcime as the typical alteration mineral, suggesting primary deposition. LA-ICP-MS zircon U-Pb dating of the tuffs yields concordant ages of 20.6 ± 0.1 Ma and 20.7 ± 0.1 Ma, indicating that the end of the Dingqinghu Fm., i.e., the Lunpola stratigraphic sequence, and the uplift of the plateau to its current high elevations occurred much later than previously estimated. The contemporaneous thick black oil shales in the Lunpola deep lake and biocoenosis together suggest that the climate was humid and warm during this period.

Paleoceanographic changes across the Latest Danian Event in the South Atlantic Ocean and planktic foraminiferal response Publication date: 1 July 2019 Source: Palaeogeography, Palaeoclimatology, Palaeoecology, Volume 525 Author(s): Sofie Jehle, André Bornemann, Anna Friederike Lägel, Arne Deprez, Robert P. Speijer Abstract A number of short warming events occurred during Paleocene and Eocene, of which the "Paleocene-Eocene Thermal Maximum" (PETM, 56 Ma) is the most severe and most investigated event. The less known "Latest Danian Event" (LDE) at 62.2 Ma represents a 200 ky-lasting warming phase, superimposed on a long-term cooling trend after the Early Paleocene. South Atlantic ODP Site 1262 data, covering ~1 myr, indicate a warming of the entire water column by 1.5–2.6 °C, accompanying a prominent negative carbon isotope excursion (~0.9–1.1‰) and a long-term re-organization of the planktic foraminiferal fauna associated with the LDE. This study unravels a different paleoceanographic evolution of the upper ocean structure compared to results from Pacific ODP Site 1210. Unlike the Pacific, the Atlantic site lacks an apparent change of stratification as well as an overall dominance of thermocline dwelling planktic foraminifera species and a low abundance of surface dwelling photosymbiotic foraminifera. Within the LDE, indications for a slightly enhanced stratification of the upper water column and transient warming were indicated when surface dwelling planktic foraminifera became temporarily more abundant. The long-term evolution in planktic foraminifera with the disappearance of Praemurica at the LDE onset and a contemporaneous rise in Morozovella is similar to the trends reported from Shatsky Rise ODP Site 1210.

Early diversification of Ordovician chitinozoans on Baltica: New data from the Jägala waterfall section, northern Estonia Publication date: 1 July 2019 Source: Palaeogeography, Palaeoclimatology, Palaeoecology, Volume 525 Author(s): Jaak Nõlvak, Yan Liang, Olle Hints Abstract Chitinozoans are a group of biostratigraphically valuable microfossils which appeared in the Tremadocian and diversified during the rest of the Great Ordovician Biodiversification Event, to become extinct in the Devonian. The early diversification of chitinozoans on the Baltica palaeocontinent has remained poorly known due to preservation and collecting bias. In this study we document the distribution of chitinozoans through the upper Tremadocian to lower Darriwilian strata in the Jägala waterfall section from northern Estonia, in order to better understand the regional diversity patterns and biogeographic links. Fifty samples were studied from the 7.5 m succession of sandstones, marls and carbonates revealing 47 chitinozoan species and 11 genera. The lowermost productive samples, attributed to the late Tremadocian, represent the earliest rich chitinozoan fauna from Baltica. Combined with previous reports this shows a diversity peak in the Tremadocian with balanced total diversity reaching 17, possibly followed by a decline in the Floian and then a rather gradual increase through the Dapingian and early Darriwilian. The new diversity curve shows more than three times higher values for the Tremadocian and Dapingian than indicated in previous studies, and thus less abrupt radiation in the Darriwilian. The Jägala section provided also new data on the regional biozonal species Conochitina cucumis and Cyathochitina regnelli, which characterize the middle and upper Volkhov and lower Kunda regional stages, respectively. Several other biostratigraphically significant taxa typical of other regions were recovered from the Jägala section, including Conochitina decipiens and Desmochitina bulla. A new short-ranging species Laufeldochitina toilaensis sp. nov. is introduced, having potential to become an index species for middle Dapingian strata. The early Darriwilian assemblage from Jägala includes Rhabdochitina sp. A with up to 2.7 mm long vesicle, making it the largest chitinozoan ever reported.

Cancer

FOXO3a knockdown promotes radioresistance in nasopharyngeal carcinoma by inducing epithelial-mesenchymal transition and the Wnt/β-catenin signaling pathway Publication date: 28 July 2019 Source: Cancer Letters, Volume 455 Author(s): Min Luo, Cheng Wu, Ergang Guo, Shan Peng, Linli Zhang, Wei Sun, Dongbo Liu, Guangyuan Hu, Guoqing Hu Abstract Mutations in the forkhead box O 3a (FOXO3a) gene are closely related to the progression of several types of cancers. However, few studies explore the relationship between FOXO3a and nasopharyngeal carcinoma (NPC). Our findings demonstrate that silencing FOXO3a promotes tumor radioresistance of NPC in vitro and in vivo through inducing EMT and activating Wnt/β-catenin signal pathway. These data establish that FOXO3a can be a novel and reliable NPC marker and a potential therapeutic target against NPC.

CD100-plexin-B1 induces epithelial-mesenchymal transition of head and neck squamous cell carcinoma and promotes metastasis Publication date: 28 July 2019 Source: Cancer Letters, Volume 455 Author(s): Chen Zhang, Hongjiang Qiao, Weinan Guo, Yuan Liu, Luting Yang, Yufeng Liu, Boquan Jin, Meng Fu, Gang Wang, Wei Li Abstract Head and neck squamous cell carcinoma (HNSCC) is one of the most lethal cancers mainly due to the high rate of metastasis. Here, we find that the expression level of CD100 in HNSCC is positively correlated with the T category, pathological grade and lymph node metastasis of the tumor. The level of soluble CD100 (sCD100) is highly increased in serum of HNSCC patients, and sCD100 markedly induces the epithelial-mesenchymal transition (EMT) of HNSCC through its receptor, Plexin-B1 (PlxnB1), and promotes the metastasis in a xenograft mouse model. Furthermore, sCD100 promotes the stabilization of Snail through the regulation of the Vav1-Rac1/RhoA-p21-activated kinase pathway for the induction of EMT. Anti-CD100 antibody abolishes the CD100-induced EMT and prevents the metastasis of HNSCC, and anti-CD100 antibody also increases the drug sensitivity of HNSCC. Taken together, our study shows for the first time that CD100 induces the EMT of HNSCC and promotes the metastasis, and CD100 would be a candidate as a novel prognostic biomarker and a potential therapeutic target for HNSCC.

FA-NBs-IR780: Novel multifunctional nanobubbles as molecule-targeted ultrasound contrast agents for accurate diagnosis and photothermal therapy of cancer Publication date: 28 July 2019 Source: Cancer Letters, Volume 455 Author(s): Yamei Shen, Wei Lv, Hengli Yang, Wenbin Cai, Ping Zhao, Li Zhang, Jian Zhang, Lijun Yuan, Yunyou Duan Abstract Early accurate diagnosis and targeted therapy for cancer are essential to improve the prognosis of patients. With the emergence of molecular imaging, molecule-targeted ultrasound imaging for the non-invasive and precise detection of cancer has attracted increased attention. The investigation of molecule-targeted ultrasound contrast agents (UCAs) with excellent performance is urgently needed. In this study, we synthetized folic acid and IR-780 on self-made nanobubbles and prepared novel UCAs, named FA-NBs-IR780. The results showed that the conjugates had a uniform size distribution (591 ± 52 nm). In vitro and in vivo experiments demonstrated that FA-NBs-IR780 can target tumour cells via dual molecular targeting, perform enhanced-contrast ultrasound imaging and near-infrared fluorescence (NIRF) imaging for the precise detection of tumours, and induce targeted photothermal therapy in lesions irradiated at 808 nm. Ex vivo experiments further confirmed that FA-NBs-IR780 efficiently induced tumour cell apoptosis and inhibited tumour growth. The newly fabricated FA-NBs-IR780 were observed to be molecule-targeted dual-mode UCAs and may have potential applications in early accurate diagnosis and targeted therapy of cancer in the future.

miR-374a-5p promotes tumor progression by targeting ARRB1 in triple negative breast cancer Publication date: 10 July 2019 Source: Cancer Letters, Volume 454 Author(s): Dasom Son, Yesol Kim, Sera Lim, Hyeok-Gu Kang, Da-Hyun Kim, Jee Won Park, Woosung Cheong, Hyun Kyung Kong, Wonshik Han, Woong-Yang Park, Kyung-Hee Chun, Jong Hoon Park Abstract Triple negative breast cancer (TNBC) has higher aggressiveness and poorer outcomes compared with other subtypes of breast cancer. However, the genomic and molecular aberrations of TNBC are largely unknown. In this study, miR-374a-5p was discovered as a novel TNBC-specific miRNA and its functions and the molecular mechanisms involved were investigated. Combined gene expression profiling of miRNA-microarray and human transcriptome dataset analysis revealed that miR-374a-5p is specifically upregulated in TNBC patients. Functional studies using in vitro and in vivo models indicated that upregulated miR-374a-5p promotes tumor progression in TNBC. miR-374a-5p was also found to directly target arrestin beta 1 (ARRB1) that is specifically downregulated in TNBC patients in several human genomic datasets. Overexpressed ARRB1 reduced TNBC cell growth and migration, and the ARRB1 expression level is inversely correlated with the histological grade of the breast cancer and positively associated with TNBC patient survival, suggestive of a tumor-suppressive function of ARRB1 in breast cancer. Interestingly, increased ARRB1 activates AMPK in TNBC cells, associated with the expression of miR-374a-5p. Taken together, the findings suggest that miR-374a-5p is a potential prognostic marker of TNBC.

Glycochenodeoxycholate promotes hepatocellular carcinoma invasion and migration by AMPK/mTOR dependent autophagy activation Publication date: 10 July 2019 Source: Cancer Letters, Volume 454 Author(s): Lu Gao, Gang Lv, Rong Li, Wen-ting Liu, Chen Zong, Fei Ye, Xiao-yong Li, Xue Yang, Jing-hua Jiang, Xiao-juan Hou, Ying-ying Jing, Zhi-peng Han, Li-xin Wei Abstract Metastasis and recurrence severely impact the treatment effect of hepatocellular carcinoma (HCC). HCC complicated with cholestasis is more prone to recurrence and metastasis. Previous studies have implicated pathogenesis of HCC by bile acid; however, the underlying mechanism is unknown yet. Glycochenodeoxycholate (GCDC) is one of most important component of bile acid (BA). In the present study, the role of GCDC in HCC cells invasion was detected by in vitro and in vivo assays. GCDC was found to significantly enhance the invasive potential of HCC cells; Further studies showed that GCDC could induce autophagy activation and higher invasive capability in HCC cells. Interestingly, inhibition of autophagy by chloroquine (CQ) reversed this phenomenon. Subsequently, the correlation between TBA expression level and clinicopathological characteristics was analyzed in HCC patients. Clinically, high TBA level in HCC tissue was found to be associated with more invasive and poor survival in HCC patients. Mechanistic study showed that bile acid induced autophagy by targeting the AMPK/mTOR pathway in HCC cells. Therefore, our results suggest that bile acid may promote HCC invasion via activation of autophagy and the level of bile acid may serve as a potential useful indicator for prognosis of HCC patients.

Disruption of oncogenic liver-intestine cadherin (CDH17) drives apoptotic pancreatic cancer death Publication date: 10 July 2019 Source: Cancer Letters, Volume 454 Author(s): Xinjian Liu, Yue Huang, Hao Yuan, Xiaoqiang Qi, Yariswamy Manjunath, Diego Avella, Jussuf T. Kaifi, Yi Miao, Min Li, Kuirong Jiang, Guangfu Li Abstract Liver–intestine cadherin (CDH17) has been known to function as a tumor stimulator and diagnostic marker for almost two decades. However, its function in highly malignant pancreatic cancer (PC) has yet to be elucidated. Using different strategies including siRNA, shRNA, and CRISPR technology, we successfully induced knockdown and knockout of CDH17 in Panc02-H7 cells and established the corresponding stable cell lines. With these cells, we demonstrated that loss of CDH17 function not only suppressed Panc02-H7 cell growth in vitro but also significantly slowed orthotopic tumor growth in vivo, resulting in the significant life extension. In vitro studies demonstrated that impairing CDH17 inhibited cell proliferation, colony formation, and motility by mechanistically modulating pro- and anti-apoptosis events in PC cells, as CDH17 suppression obviously increased expression of Bad, cytochrome C, cleaved caspase 3, and cleaved PARP, and reduced expression of Bcl-2, Survivin, and pAkt. In vivo studies showed CDH17 knockout resulted in apoptotic PC tumor death through activating caspase-3 activity. Taken together, CDH17 functions as an oncogenic molecule critical to PC growth by regulating tumor apoptosis signaling pathways and CDH17 could be targeted to develop an anti-PC therapeutic approach.

ZY0511, a novel, potent and selective LSD1 inhibitor, exhibits anticancer activity against solid tumors via the DDIT4/mTOR pathway Publication date: 10 July 2019 Source: Cancer Letters, Volume 454 Author(s): Yan Li, Lei Tao, Zeping Zuo, Yang Zhou, Xinying Qian, Yiyun Lin, Hui Jie, Chunqi Liu, Zhuoling Li, Huaqin Zhang, Hu Zhang, Xiaobo Cen, Shengyong Yang, Yinglan Zhao Abstract Lysine-specific demethylase1 (LSD1) plays a crucial role in cancer and has become a promising target for cancer therapy. However, the mechanism underlying the role of LSD1 in oncogenesis is poorly understood, and more effective LSD1 inhibitors are needed. Here we report the biological activity of a novel LSD1 inhibitor named ZY0511. ZY0511 specifically inhibited LSD1 activity and the proliferation of various human cancer cells especially the HeLa and HCT116 cells. ZY0511 significantly increased the expression of DDIT4, a known mTORC1 suppressor, which was a direct downstream target of LSD1 confirmed by ChIP-PCR. ZY0511-induced LSD1 inhibition upregulated the expression of DDIT4 by altering histone H3K4 methylation levels at its promoter, thus suppressing mTORC1 activity. Knockdown of DDIT4 attenuated the anticancer effect of ZY0511. Intraperitoneal administration of ZY0511 significantly prevented the growth of HCT116 and HeLa xenografts in mice and showed no detectable toxicity. Moreover, DDIT4 expression was correlated with the sensitivity of human cancer cells to chemotherapy. Taken together, ZY0511 showed therapeutic potential for solid tumors, the induction of DDIT4 may be used as a predictive biomarker of LSD1 inhibitors.

Long non-coding RNA HULC activates HBV by modulating HBx/STAT3/miR-539/APOBEC3B signaling in HBV-related hepatocellular carcinoma Publication date: 10 July 2019 Source: Cancer Letters, Volume 454 Author(s): Yunxia Liu, Jinyan Feng, Mingming Sun, Guang Yang, Hongfeng Yuan, Yinqi Wang, Yanan Bu, Man Zhao, Shuqin Zhang, Xiaodong Zhang Abstract Long noncoding RNA HULC is identified and highly expressed in hepatocellular carcinoma (HCC). Hepatitis B virus (HBV) is a key driver of liver cancer. In the present study, we found that HULC remarkably elevated the levels of HBeAg, HBsAg, HBcAg, pgRNA, HBx, HBV DNA and covalently closed circular DNA (cccDNA), which activated the HBV replication in HBV-expressing hepatoma cells or de novo HBV-infected cell lines (PHH, HepG2-NTCP and dHepaRG). Mechanistically, HULC enhanced HBV cccDNA stability by down-regulating the APOBEC3B in hepatoma cells. HULC significantly up-regulated microRNA-539, which targeted the 3′UTR of APOBEC3B mRNA. Luciferase reporter gene assays revealed a putative STAT3-binding site located in the upstream of miR-539 promoter. Moreover, we identified that HULC was able to elevate HBx, which co-activated the STAT3 to stimulate the miR-539 promoter. Then, miR-539 down-regulated APOBEC3B and promoted HBV replication. Functionally, HULC enhanced the growth of hepatoma cells by activating HBV in vitro and in vivo, which could be blocked by overexpressing APOBEC3B. In conclusion, HULC activates HBV by modulating HBx/STAT3/miR-539/APOBEC3B signaling in HBV-related HCC.

IL1RN mediates the suppressive effect of methionine deprivation on glioma proliferation Publication date: 10 July 2019 Source: Cancer Letters, Volume 454 Author(s): Kaikai Wang, Huailei Liu, Jiaqi Liu, Xiaoxiong Wang, Lei Teng, Jun Zhang, Yi Liu, Yizheng Yao, Jun Wang, Yuan Qu, Xin Chen, Fei Peng, Hongbo Liu, Ning Wang, Yingqiang Zhong, Xu Hou, Haiping Jiang, Ozal Beylerli, Xiang Liao, Xinjian Zhang Abstract Metabolic abnormality is one of the hallmarks of cancer cells, and limiting material supply is a potential breakthrough approach for cancer treatment. Increasing researchers have been involved in the study of glioma cell metabolism reprogramming since the significance of IDH1 was confirmed in glioma. However, the molecular mechanisms underlying metabolic reprogramming induced by methionine deprivation regulates glioma cell proliferation remain unclear. Here we demonstrated that methionine deprivation inhibited glioma cell proliferation via downregulating interleukin 1 receptor antagonist (IL1RN) both in vitro and in vivo, methionine deprivation or knocking down IL1RN induced glioma cell cycle arrest. Moreover, we confirmed that IL1RN is a tumor associated gene and its expression is negatively correlated with the survival time of glioma patients. Altogether these results demonstrate a strong rationale insight that targeting amino acid metabolism such as methionine deprivation/IL1RN related gene therapy may offer novel direction for glioma treatment.

Combined class I histone deacetylase and mTORC1/C2 inhibition suppresses the initiation and recurrence of oral squamous cell carcinomas by repressing SOX2 Publication date: 10 July 2019 Source: Cancer Letters, Volume 454 Author(s): Xueyi Liang, Miao Deng, Chi Zhang, Fan Ping, Hongfei Wang, Yun Wang, Zhaona Fan, Xianyue Ren, Xiaoan Tao, Tong Wu, Jian Xu, Bin Cheng, Juan Xia Abstract Treatment of oral squamous cell carcinoma (OSCC) remains a challenge because of the lack of effective early treatment strategies and high incidence of relapse. Here, we showed that combined 4SC-202 (a novel selective class I HDAC inhibitor) and INK128 (a selective mTORC1/C2 inhibitor) treatment exhibited synergistic effects on inhibiting cell growth, sphere-forming ability, subcutaneous tumor formation and ALDH1+ cancer stem cells (CSCs) in OSCC. The initiation of OSCC was significantly inhibited by combined treatment in 4NQO-induced rat model. In addition, upregulated SOX2 was associated with advanced and metastatic tumors in OSCC patients and was responsible for the drug-resistance property of OSCC cells. The inhibitory effect of combined treatment on cell viability and ALDH1+ CSCs were attenuated by SOX2 verexpression. Furthermore, combined treatment can effectively overcome chemoresistance and inhibit the growth of recurrent OSCC in vitro and in vivo. Mechanistically, 4SC-202 and INK128 repressed SOX2 expression through miR-429/miR-1181-mediated mRNA degradation and preventing cap-dependent mRNA translation, respectively. These results suggest that combined class I histone deacetylase and mTORC1/C2 inhibition suppresses the carcinogenesis and recurrence of OSCC by repressing SOX2. Graphical abstract