bluuebunny/crossref_metadata_2025 huggingface dataset:
6,7‐Benzotropolone Syntheses Based on Ring‐Closing Metatheses and Four‐Electron Oxidations
Authors: Kreibich Michael , Gemander Manuel , Peter David , Yadav Dharmendra B. , de Koning Charles B. , Fernandes Manuel A. , Green Ivan R. , van Otterlo Willem A. L. , Brückner Reinhard
Published: March 2020

Four homoallyl ortho‐vinylaryl ketones (10a‐d) – 1,8‐dienes of sorts – were prepared by several approaches. In the presence of 1–2 mol‐% Grubbs‐II catalyst, they ring‐closed to give 6,7‐dihydrobenzocyclohepten‐5‐ones (11a‐d) in 90–96 % yield. With SeO2 the parent compound (11a) delivered benzocyclohepten‐5‐one (13a) and/or selenium‐containing compounds (18–22) but no more than traces of 6,7‐benzotropolone (5a). However, 5a was accessible from compound 11a via the sodium enolate and allowing it to react with a stream of oxygen (43 % yield). The sodium enolates of the substituted 6,7‐dihydrobenzocyclohepten‐5‐ones 11b–d and oxygen underwent analogous 4‐electron oxidations. This furnished the substituted 6,7‐benzotropolones 11b‐d. In contrast, the corresponding lithium enolates were inert towards oxygen. The 6,7‐dihydrobenzocyclohepten‐5‐one 11d was also accessed differently, namely by a Grubbs‐II catalyst‐mediated RCM/C=C migration tandem reaction of the allyl ortho‐allylaryl ketone 73 – another 1,8‐diene of sorts (90 % yield).
Pain modulation by illusory body rotation: A new way to disclose the interaction between the vestibular system and pain processing
Authors: Daniel Aster , Barker Leon , Martini Matteo
Published: March 2020

Background

Clinical and experimental evidence advocates a structural and functional link between the vestibular and other sensory systems. For instance, visuo‐vestibular and vestibular–somatosensory interactions have been widely reported. However, whether visual inputs carrying vestibular information can modulate pain is not yet clear. Recent evidence using natural vestibular stimulation or moving visual stimuli, points at an unspecific effect of distraction.

Methods

By using immersive virtual reality (VR), we created a new way to prompt the vestibular system through the vision of static visual cues, studying the possible interaction with pain. Twenty‐four healthy participants were visually immersed in a virtual room which could appear with five different degrees of rotation in the sagittal axis, either towards the right, left or with no rotation. Participants' heat pain thresholds and subjective reports of perceived body rotation, sense of presence and attention were measured.

Results

‘Being’ in a tilted room induced the sensation of body rotation in our participants, even though they were always in an upright position. We also found that rotating the visual scenario can modulate the participants' pain thresholds, determining a significant increase when a left tilt is displayed. In addition, a positive correlation between the perceived body midline rotation and pain threshold was found when the virtual room was titled 15 degrees toward the left. Importantly, all VR conditions were found to be equally distractive.

Conclusions

Vestibular information present in static visual cues can modulate experimentally‐induced acute pain according to a side‐dependent manner and bypassing supramodal attentional mechanisms. These findings may help refining pain management approaches based on multimodal stimulation.

Significance

This study explored how the visualization of static environments in immersive virtual reality can lead to pain threshold modulation through the activation of the vestibular system. Immersion into rotated virtual environments led to the illusory sensation of body rotation, and this sensation was found to be related with a modulation of pain perception. Possible analgesic effects due to distraction could be ruled out. These results expand our current knowledge about how the visual, vestibular and somatosensory (pain) systems interact. These findings may influence future pain treatment strategies based on multisensory stimulation.
Minocycline reduces experimental muscle hyperalgesia induced by repeated nerve growth factor injections in humans: A placebo‐controlled double‐blind drug‐crossover study
Authors: Dunn James S. , Nagi Saad S. , Mahns David A.
Published: March 2020

Background

Hyperalgesia is a heightened pain response to a noxious stimulus and is a hallmark of many common neuropathic and chronic pain conditions. In a double‐blind placebo‐controlled drug‐crossover trial, the effects of concomitant and delayed minocycline treatment on the initiation and resolution of muscle hyperalgesia were tested.

Methods

An initial cohort (n = 10) received repeated injections (5 µg: days 0, 2 and 4) of nerve growth factor (NGF) in the flexor carpi ulnaris muscle of the forearm and pressure pain thresholds were collected at day 0 (control), day 7 (peak) and day 14 (recovery). A second cohort (n = 18) underwent an identical procedure, however, half received a placebo between days 0 and 7 before switching to minocycline from days 7 to 14 (P1/M2), while the remaining subjects received minocycline (day 0: 200mg then 100mg b.i.d. for 7 days) before switching to placebo (M1/P2).

Results

The initial cohort exhibited a diffuse muscular pain hypersensitivity with a decrease in pressure pain thresholds at day 7 before a partial return to normalcy at day 14. The P1/M2 treatment group exhibited an identical peak in hypersensitivity at day 7, however, after switching to minocycline in week 2 showed a significant reduction in muscle hyperalgesia compared with the initial cohort at day 14. The M1/P2 treatment group had significantly less (~43%) hyperalgesia at day 7 compared with the other groups.

Conclusions

The study indicates that the administration of minocycline can reduce experimentally induced muscle pain regardless of the time of administration.

Significance

In a double‐blind placebo‐controlled drug‐crossover study, the common antibiotic minocycline was found to reduce the muscle hyperalgesia induced by intramuscular injection of nerve growth factor. The results of the study showed that both concomitant (pre‐emptive) and delayed administration of minocycline can ameliorate the onset and facilitate the resolution of experimentally induced muscle hyperalgesia.
Effectiveness of percutaneous electrical nerve stimulation for musculoskeletal pain: A systematic review and meta‐analysis
Authors: Plaza‐Manzano Gustavo , Gómez‐Chiguano Guido F. , Cleland Joshua A. , Arías‐Buría Jose L. , Fernández‐de‐las‐Peñas César , Navarro‐Santana Marcos J.
Published: March 2020

Background and Objective

To evaluate the effects of percutaneous electrical stimulation (PENS) alone or as an adjunct with other interventions on pain and related disability in musculoskeletal pain conditions.

Databases and Data Treatment

Search of MEDLINE, EMBASE, AMED, CINAHL, EBSCO, PubMed, PEDro, Cochrane Library, SCOPUS and Web of Science databases. Randomized controlled trials where at least one group received any form of PENS for musculoskeletal condition. Studies had to include humans and collect outcomes on pain and related disability in musculoskeletal pain. Risk of bias was assessed by the Cochrane Guidelines, the quality of evidence by using the GRADE approach. Standardized mean differences (SMD) were calculated.

Results

Sixteen studies were included and included heterogeneous musculoskeletal conditions with short‐ or midterm follow‐ups. PENS alone had a large effect (SMD −1.22, 95% CI −1.66 to −0.79) on pain and a small effect (SMD −0.33, 95% CI −0.61 to −0.06) on related disability at short‐term as compared with sham. A moderate effect of PENS alone (SMD −0.71, 95% CI −1.23 to −0.19) on pain when compared with other interventions was observed. The inclusion of PENS with other interventions had a moderate effect for decreasing pain at short‐ (SMD −0.70, 95% CI −1.02 to −0.37) and midterm (SMD −0.68, 95% CI −1.10 to −0.27). No effect at midterm (SMD −0.21, 95% CI −0.52 to 0.10) on related disability was seen. The risk of bias was generally low; but the heterogenicity of the results downgraded the level of evidence.

Conclusion

There is low level of evidence suggesting the effects of PENS alone or in combination for pain, but not related disability, in musculoskeletal pain.

Level of Evidence

Therapy, level 1a.

Registration number: CRD42019131331.

Significance

This meta‐analysis investigating the effectiveness of PENS for the management of pain and related disability in musculoskeletal pain conditions found that PENS could decrease level of pain intensity but not relateddisability in musculoskeletal pain disorders.
Preliminary design and evaluation of large‐diameter superconducting cable toward GW‐class hybrid energy transfer of electricity, liquefied natural gas, and liquefied nitrogen
Authors: Chen Yu , Jiang Shan , Chen Xiao Yuan , Wang Ya Fang , Li Tao
Published: March 2020

Considering the temperature rises caused by the friction loss and heat leakage, long‐distance and high‐capacity transportation of liquefied natural gas (LNG) is currently not very feasible by using a cryogenic pipeline. As a fire‐proof and pollution‐free liquid, liquefied nitrogen (LN2) is introduced to cool the LNG pipe and high temperature superconducting (HTS) cable simultaneously for exploring a new hybrid energy transfer (HET) concept of the LNG, LN2, and electricity. In this new concept, both the LNG pipe and HTS cable are inserted into the same LN2 pipe, and these two coaxial assemblies are designed to have very large diameters for enhancing the energy transfer capacity. As case studies, two HTS cables rated at 2 kA and 10 kA are structurally optimized by using a particle swarm optimization method. Two performance indicators of required tape length and generated AC loss per meter cable are introduced to evaluate the capital cost and operating cost with thousands of optimized parameters. The results show that this new HET concept has significant potentials to avoid excessive LNG temperature rise and reduce the HTS tape and cooling costs, and thus lays some bases for the use in future power and energy transfer applications.
Effects of Herbicides on Flowering
Authors: Carpenter David J. , Mathiassen Solvejg K. , Boutin Céline , Strandberg Beate , Casey Carlene S. , Damgaard Christian
Published: March 2020

Herbicides have been shown to reduce flower production and to delay flowering, with results varying among herbicides and tested plant species. We investigated the effects of herbicides on flowering in an extensive greenhouse study conducted in Canada and Denmark. The effects of low doses of 5 different herbicides (bromoxynil, ioxynil + bromoxynil, metsulfuron-methyl, clopyralid, and glyphosate), simulating realistic drift scenarios (1 and 5% recommended field rates), on plant flowering were examined using 9 wild plant species exposed at either the seedling (6- to 8-leaf) or flower bud stage. Following herbicide exposure, initial flowering date as well as flower production over time were recorded over the growing period. The effect of herbicides on cumulative flower numbers and flowering time were modeled using Gompertz growth models. Significant delays to peak flowering and/or reductions in flower production were observed in at least one plant species for all tested herbicides, with glyphosate often exhibiting the greatest negative effects, that is, plant death. Except for ioxynil + bromoxynil, there was no clear evidence of either the seedling or the flower bud stage being more sensitive. Overall, 58% of all species × life stage × herbicide treatments resulted in either a statistically significant or a strong decline in flower production with herbicide application rates up to 5% of recommended field rates, whereas significant or strong delays in peak flowering were also detected but were slightly less common. Effects at 1% label rates were minimal. Simultaneous delays to peak flowering and reductions in total flower production occurred in approximately 25% of all cases, indicating that herbicide application rates simulating realistic drift scenarios would likely have negative effects on wild floral communities. Environ Toxicol Chem 2020;39:1244–1256. © 2020 SETAC
Cryptic aberrations may allow more accurate prognostic classification of patients with myelodysplastic syndromes and clonal evolution
Authors: Svobodova Karla , Lhotska Halka , Hodanova Lucie , Pavlistova Lenka , Vesela Denisa , Belickova Monika , Vesela Jitka , Brezinova Jana , Sarova Iveta , Izakova Silvia , Lizcova Libuse , Siskova Magda , Jonasova Anna , Cermak Jaroslav , Michalova Kyra , Zemanova Zuzana
Published: March 2020

The karyotype of bone‐marrow cells at the time of diagnosis is one of the most important prognostic factors in patients with myelodysplastic syndromes (MDS). In some cases, the acquisition of additional genetic aberrations (clonal evolution [CE]) associated with clinical progression may occur during the disease. We analyzed a cohort of 469 MDS patients using a combination of molecular cytogenomic methods to identify cryptic aberrations and to assess their potential role in CE. We confirmed CE in 36 (8%) patients. The analysis of bone‐marrow samples with a combination of cytogenomic methods at diagnosis and after CE identified 214 chromosomal aberrations. The early genetic changes in the diagnostic samples were frequently MDS specific (17 MDS‐specific/57 early changes). Most progression‐related aberrations identified after CE were not MDS specific (131 non‐MDS‐specific/155 progression‐related changes). Copy number neutral loss of heterozygosity (CN‐LOH) was detected in 19% of patients. MDS‐specific CN‐LOH (4q, 17p) was identified in three patients, and probably pathogenic homozygous mutations were found in TET2 (4q24) and TP53 (17p13.1) genes. We observed a statistically significant difference in overall survival (OS) between the groups of patients divided according to their diagnostic cytogenomic findings, with worse OS in the group with complex karyotypes (P = .021). A combination of cytogenomic methods allowed us to detect many cryptic genomic changes and identify genes and genomic regions that may represent therapeutic targets in patients with progressive MDS.
Geochemistry and palynology of the upper Albian at the Abu Gharadig Basin, southern Tethys: Constraints on the oceanic anoxic event 1d
Authors: Mansour Ahmed , Gentzis Thomas , Carvajal‐Ortiz Humberto , Tahoun Sameh S. , Wagreich Michael
Published: March 2020

The mid‐Cretaceous period was a time of abnormal polar temperature warming and consequent global oceanic anoxia that enhanced the widespread accumulation of organic matter‐rich marine sediments. The current contribution focuses on the late Albian oceanic anoxic subevent 1d (OAE 1d) that represents a period of increased perturbation of the global carbon cycle. Palynological and geochemical investigations, including TOC/Rock‐Eval pyrolysis and stable carbon isotope of organic matter (δ13CTOC), were conducted for the upper part of the Kharita Formation from the Abu Gharadig Basin, north Western Desert, Egypt, to (a) confirm the age of this unit, (b) assess the depositional environments and prevalent redox conditions, and (c) confirm the occurrence of the OAE 1d within the sedimentary record in this part of the southern Tethys. Based on the TOC content and Rock‐Eval pyrolysis parameters, the organic matter in the upper Kharita Formation is expected to have a limited hydrocarbon generation potential consisting of gas only. The sedimentation patterns and role of continental weathering and runoff influx were assessed through trace element ratios and relationships. Redox conditions were interpreted based on Stotal–TOC relationship, Stotal–TOC–Fetotal ternary plot, Rock‐Eval S2, trace element proxies and relationships (including Cr vs. V + Ni, V/Cr vs. Stotal, and Stotal/Fe vs. Mn/Stotal). The ratio of TOC/Ptotal was implemented to assess both marine primary productivity and oceanic anoxia during this period. Biostratigraphic constraints showed that the upper part of the Kharita Formation spanned a late Albian age (Cretacaeiporites densimurus Interval Zone). Results showed that the OAE 1d was characterized by organic matter deposition during dysoxic‐suboxic to anoxic redox conditions in a fluvio‐deltaic to shallow marine depositional environment in this part of the Tethys. A low to moderate primary productivity was also prevalent during a fairly high rate of sediment accumulation. Regional correlation of positive δ13CTOC excursion during the OAE 1d throughout the Tethys and Atlantic oceans matched well, in several parts, with the current study interval that confirm the global distribution of the OAE 1d. In this region of the southern Tethys, an overall collapse of water column stratification during the OAE 1d principally related to decreased terrestrial input at low relative sea level, as evidenced from freshwater algae Pediastrum and pteridophyte spore Crybelosporites pannuceus; however, orbital forcing likely induced monsoonal activity at low‐latitudes in the SE, N, and W Tethys.
Functional connectivity of the frontotemporal network in preattentive detection of abstract changes: Perturbs and observes with transcranial magnetic stimulation and event‐related optical signal
Authors: Xiao Xue‐Zhen , Shum Yu‐Hei , Lui Troby K.‐Y. , Wang Yang , Cheung Alexandra T.‐C. , Chu Winnie C. W. , Neggers Sebastiaan F. W. , Chan Sandra S.‐M. , Tse Chun‐Yu
Published: March 2020

Current theories of automatic or preattentive change detection suggest a regularity or prediction violation mechanism involving functional connectivity between the inferior frontal cortex (IFC) and the superior temporal cortex (STC). By disrupting the IFC function with transcranial magnetic stimulation (TMS) and recording the later STC mismatch response with event‐related optical signal (EROS), previous study demonstrated a causal IFC‐to‐STC functional connection in detecting a pitch or physical change. However, physical change detection can be achieved by memory comparison of the physical features and may not necessarily involve regularity/rule extraction and prediction. The current study investigated the IFC–STC functional connectivity in detecting rule violation (i.e., an abstract change). Frequent standard tone pairs with a constant relative pitch difference, but varying pitches, were presented to establish a pitch interval rule. This abstract rule was violated by deviants with reduced relative pitch intervals. The EROS STC mismatch response to the deviants was abolished by the TMS applied at the IFC 80 ms after deviance onset, but preserved in the spatial (TMS on vertex), auditory (TMS sound), and temporal (200 ms after deviance onset) control conditions. These results demonstrate the IFC–STC connection in preattentive abstract change detection and support the regularity or prediction violation account.
Genetic Pathways in Nonalcoholic Fatty Liver Disease: Insights From Systems Biology
Authors: Sookoian Silvia , Pirola Carlos J. , Valenti Luca , Davidson Nicholas O.
Published: March 2020

Nonalcoholic fatty liver disease (NAFLD) represents a burgeoning worldwide epidemic whose etiology reflects multiple interactions between environmental and genetic factors. Here, we review the major pathways and dominant genetic modifiers known to be relevant players in human NAFLD and which may determine key components of the heritability of distinctive disease traits including steatosis and fibrosis. In addition, we have employed general assumptions which are based on known genetic factors in NAFLD to build a systems biology prediction model that includes functional enrichment. This prediction model highlights additional complementary pathways that represent plausible intersecting signaling networks that we define here as an NAFLD‐Reactome. We review the evidence connecting variants in each of the major known genetic modifiers (variants in patatin‐like phospholipase domain containing 3, transmembrane 6 superfamily member 2, membrane‐bound O‐acyltransferase domain containing 7, glucokinase regulator, and hydroxysteroid 17‐beta dehydrogenase 13) to NAFLD and expand the associated underlying mechanisms using functional enrichment predictions, based on both preclinical and cell‐based experimental findings. These major candidate gene variants function in distinct pathways, including substrate delivery for de novo lipogenesis; mitochondrial energy use; lipid droplet assembly, lipolytic catabolism, and fatty acid compartmentalization; and very low‐density lipoprotein assembly and secretion. The NAFLD‐Reactome model expands these pathways and allows for hypothesis testing, as well as serving as a discovery platform for druggable targets across multiple pathways that promote NAFLD development and influence several progressive outcomes. In conclusion, we summarize the strengths and weaknesses of studies implicating selected variants in the pathophysiology of NAFLD and highlight opportunities for future clinical research and pharmacologic intervention, as well as the implications for clinical practice.
Cell‐Free Virus‐Host Chimera DNA From Hepatitis B Virus Integration Sites as a Circulating Biomarker of Hepatocellular Cancer
Authors: Li Chiao‐Ling , Ho Ming‐Chih , Lin You‐Yu , Tzeng Sheng‐Tai , Chen Yun‐Ju , Pai Hsin‐Yung , Wang Ya‐Chun , Chen Chi‐Ling , Lee Yu‐Hsin , Chen Ding‐Shinn , Yeh Shiou‐Hwei , Chen Pei‐Jer
Published: March 2020

Background and Aims

Early recurrence of hepatocellular carcinoma (HCC) after surgical resection compromises patient survival. Timely detection of HCC recurrence and its clonality is required to implement salvage therapies appropriately. This study examined the feasibility of virus‐host chimera DNA (vh‐DNA), generated from junctions of hepatitis B virus (HBV) integration in the HCC chromosome, as a circulating biomarker for this clinical setting.

Approach and Results

HBV integration in 50 patients with HBV‐related HCC was determined by the Hybridization capture‐based next‐generation sequencing (NGS) platform. For individual HCC, the vh‐DNA was quantified by specific droplet digital PCR (ddPCR) assay in plasma samples collected before and 2 months after surgery. HBV integrations were identified in 44 out of 50 patients with HBV‐related HCC. Tumor‐specific ddPCR was developed to measure the corresponding vh‐DNA copy number in baseline plasma from each patient immediately before surgery. vh‐DNA was detected in 43 patients (97.7%), and the levels correlated with the tumor sizes (detection limit at 1.5 cm). Among the plasma collected at 2 months after surgery, 10 cases (23.3%) still contained the same signature vh‐DNA detected at baseline, indicating the presence of residual tumor cells. Nine of them (90%) experienced HCC recurrence within 1 year, supporting vh‐DNA as an independent risk factor in predicting early recurrence. Analysis of circulating vh‐DNA at recurrence further helped identify the clonal origin. A total of 81.8% of recurrences came from original HCC clones sharing the same plasma vh‐DNA, whereas 18.2% were from de novo HCC.

Conclusions

vh‐DNA was shown to be a circulating biomarker for detecting the tumor load in majority of patients with HBV‐related HCC and aided in monitoring residual tumor and recurrence clonality after tumor resection.
Annexin A6 Is Critical to Maintain Glucose Homeostasis and Survival During Liver Regeneration in Mice
Authors: Alvarez‐Guaita Anna , Blanco‐Muñoz Patricia , Meneses‐Salas Elsa , Wahba Mohamed , Pollock Abigail H. , Jose Jaimy , Casado Mercedes , Bosch Marta , Artuch Rafael , Gaus Katharina , Lu Albert , Pol Albert , Tebar Francesc , Moss Stephen E. , Grewal Thomas , Enrich Carlos , Rentero Carles
Published: March 2020

Background and Aims

Liver regeneration requires the organized and sequential activation of events that lead to restoration of hepatic mass. During this process, other vital liver functions need to be preserved, such as maintenance of blood glucose homeostasis, balancing the degradation of hepatic glycogen stores, and gluconeogenesis (GNG). Under metabolic stress, alanine is the main hepatic gluconeogenic substrate, and its availability is the rate‐limiting step in this pathway. Na+‐coupled neutral amino acid transporters (SNATs) 2 and 4 are believed to facilitate hepatic alanine uptake. In previous studies, we demonstrated that a member of the Ca2+‐dependent phospholipid binding annexins, Annexin A6 (AnxA6), regulates membrane trafficking along endo‐ and exocytic pathways. Yet, although AnxA6 is abundantly expressed in the liver, its function in hepatic physiology remains unknown. In this study, we investigated the potential contribution of AnxA6 in liver regeneration.

Approach and Results

Utilizing AnxA6 knockout mice (AnxA6−/−), we challenged liver function after partial hepatectomy (PHx), inducing acute proliferative and metabolic stress. Biochemical and immunofluorescent approaches were used to dissect AnxA6−/− mice liver proliferation and energetic metabolism. Most strikingly, AnxA6−/− mice exhibited low survival after PHx. This was associated with an irreversible and progressive drop of blood glucose levels. Whereas exogenous glucose administration or restoration of hepatic AnxA6 expression rescued AnxA6−/− mice survival after PHx, the sustained hypoglycemia in partially hepatectomized AnxA6−/− mice was the consequence of an impaired alanine‐dependent GNG in AnxA6−/− hepatocytes. Mechanistically, cytoplasmic SNAT4 failed to recycle to the sinusoidal plasma membrane of AnxA6−/− hepatocytes 48 hours after PHx, impairing alanine uptake and, consequently, glucose production.

Conclusions

We conclude that the lack of AnxA6 compromises alanine‐dependent GNG and liver regeneration in mice.
Combined use of cytology, p16 immunostaining and genotyping for triage of women positive for high‐risk human papillomavirus at primary screening
Authors: Cuzick Jack , Adcock Rachael , Carozzi Francesca , Gillio‐Tos Anna , De Marco Laura , Del Mistro Annarosa , Frayle Helena , Girlando Salvatore , Sani Cristina , Confortini Massimo , Zorzi Manuel , Giorgi‐Rossi Paolo , Rizzolo Raffaella , Ronco Guglielmo ,   the New Technologies for Cervical Cancer Screening Working Group
Published: March 2020

Human papillomavirus (HPV) testing is very sensitive for primary cervical screening but has low specificity. Triage tests that improve specificity but maintain high sensitivity are needed. Women enrolled in the experimental arm of Phase 2 of the New Technologies for Cervical Cancer randomized controlled cervical screening trial were tested for high‐risk HPV (hrHPV) and referred to colposcopy if positive. hrHPV‐positive women also had HPV genotyping (by polymerase chain reaction with GP5+/GP6+ primers and reverse line blotting), immunostaining for p16 overexpression and cytology. We computed sensitivity, specificity and positive predictive value (PPV) for different combinations of tests and determined potential hierarchical ordering of triage tests. A number of 1,091 HPV‐positive women had valid tests for cytology, p16 and genotyping. Ninety‐two of them had cervical intraepithelial neoplasia grade 2+ (CIN2+) histology and 40 of them had CIN grade 3+ (CIN3+) histology. The PPV for CIN2+ was >10% in hrHPV‐positive women with positive high‐grade squamous intraepithelial lesion (61.3%), positive low‐grade squamous intraepithelial lesion (LSIL+) (18.3%) and positive atypical squamous cells of undetermined significance (14.8%) cytology, p16 positive (16.7%) and, hierarchically, for infections by HPV33, 16, 35, 59, 31 and 52 (in decreasing order). Referral of women positive for either p16 or LSIL+ cytology had 97.8% sensitivity for CIN2+ and women negative for both of these had a 3‐year CIN3+ risk of 0.2%. Similar results were seen for women being either p16 or HPV16/33 positive. hrHPV‐positive women who were negative for p16 and cytology (LSIL threshold) had a very low CIN3+ rate in the following 3 years. Recalling them after that interval and referring those positive for either test to immediate colposcopy seem to be an efficient triage strategy. The same applies to p16 and HPV16.
Proteasome inhibitors and Smac mimetics cooperate to induce cell death in diffuse large B‐cell lymphoma by stabilizing NOXA and triggering mitochondrial apoptosis
Authors: Dietz Anna , Dalda Nahide , Zielke Svenja , Dittmann Jessica , van Wijk Sjoerd J.L. , Vogler Meike , Fulda Simone
Published: March 2020

Copy number gains and increased expression levels of cellular Inhibitor of Apoptosis protein (cIAP)1 and cIAP2 have been identified in primary diffuse large B‐cell lymphoma (DLBCL) tissues. Second mitochondria‐derived activator of caspases (Smac) mimetics were designed to antagonize IAP proteins. However, since their effect as single agents is limited, combination treatment represents a strategy for their clinical development. Therefore, we investigated the Smac mimetic BV6 in combination with proteasome inhibitors and analyzed the molecular mechanisms of action. We discovered that BV6 treatment sensitizes DLBCL cells to proteasome inhibition. We show a synergistic decrease in cell viability and induction of apoptosis by BV6/Carfilzomib (CFZ) treatment, which was confirmed by calculation of combination index (CI) and Bliss score. BV6 and CFZ acted together to trigger activation of BAX and BAK, which facilitated cell death, as knockdown of BAX and BAK significantly reduced BV6/CFZ‐mediated cell death. Activation of BAX and BAK was accompanied by loss of mitochondrial membrane potential (MMP) and activation of caspases. Pretreatment with the caspase inhibitor N‐benzyloxycarbonyl‐Val‐Ala‐Asp‐fluoromethylketone (zVAD.fmk) rescued BV6/CFZ‐induced cell death, confirming caspase dependency. Treatment with CFZ alone or in combination with BV6 caused accumulation of NOXA, which was required for cell death, as gene silencing by siRNA or Clustered Regularly Interspaced Short Palindromic Repeats (CRISPR)/Cas9‐mediated NOXA inactivation inhibited BV6/CFZ‐induced cell death. Together, these experiments indicate that BV6 and CFZ cooperatively induce apoptotic cell death via the mitochondrial pathway. These findings emphasize the role of Smac mimetics for sensitizing DLBCL cells to proteasome inhibition with important implications for further (pre)clinical studies.
Non‐clinical single‐ and repeated‐dose toxicity studies of ET‐26 hydrochloride in rats
Authors: Zhang YiNan , Zhang YuJun , Deng ChaoYi , Gong DeYing , Kang Yi , Liu Jin , Zhang WenSheng
Published: March 2020

ET‐26 hydrochloride (ET‐26HCl), a novel analog of etomidate, induces as effective sedation, with good cardiac and respiratory stability, as etomidate but with mild adrenocortical suppression. The objective of this study was to evaluate the potential adverse effects of ET‐26HCl in rats. In a single‐dose toxicity study, abnormal urine color (red) was observed in all groups: control (100%), 8 mg/kg (10%), 16 mg/kg (50%), and 20 mg/kg (70%) ET‐26HCl, which returned to normal on the day of dosing. There were no mortalities or serious toxicological signs; the maximum tolerable dose of ET‐26HCl was 20 mg/kg. In the repeated‐dose toxicity study, deaths occurred in the 12‐ (13.33% of males) and 16‐mg/kg/day (20% of males and 3.33% of females) groups. Abnormal urine color (red or brown) was detected in the control group (10%) and all treatment groups (30%, 46.67%, and 40% at 8, 12 and 16 mg/kg/day, respectively), at a frequency of 1.43% in the control group, 4.76% in 8 mg/kg/day, 7.62% in 12 mg/kg/day, and 4.29% in 16 mg/kg/day. Increases in neutrophils and plasma fibrinogen were temporary and recoverable effects. Macroscopic and histopathologic changes were found only at the injection sites: abnormal skin color, scabbing, thrombus, ulceration, and inflammation. During the recovery period, there was evidence of reversibility, including fibroblast proliferation and vessel recanalization. The no‐observed‐adverse‐effect level of ET‐26HCl was 8 mg/kg/day. Toxicokinetic variables of ET‐26HCl, except the calculated initial concentration in females on Day 1, showed a dose‐dependent increase to exposure, with no gender difference and no evidence of accumulation.
Nanobiocomposite based on natural polyelectrolytes for bone regeneration
Authors: Belluzo M. Soledad , Medina Lara F. , Molinuevo M. Silvina , Cortizo M. Susana , Cortizo Ana M.
Published: March 2020

We developed a composite hydrogel based on chitosan and carboxymethyl cellulose with nanometric hydroxyapatite (nHA) as filler (ranging from 0.5 to 5%), by ultrasonic methodology to be used for bone regeneration. The 3D porous‐structure of the biocomposite scaffolds were confirmed by Scanning Electron Microscopy and Microtomography analysis. Infrared analysis did not show specific interactions between the organic components of the composite and nHA in the scaffold. The hydrogel properties of the matrices were studied by swelling and mechanical tests, indicating that the scaffold presented a good mechanical behavior. The degradation test demonstrated that the material is slowly degraded, while the addition of nHA slightly influences the degradation of the scaffolds. Biocompatibility studies carried out with bone marrow mesenchymal progenitor cells (BMPC) showed that cell proliferation and alkaline phosphatase activity were increased depending on the matrix nHA content. On the other hand, no cytotoxic effect was observed when RAW264.7 cells were seeded on the scaffolds. Altogether, our results allow us to conclude that these nanobiocomposites are promising candidates to induce bone tissue regeneration.
Bioactive micropatterning of biomaterials for induction of endothelial progenitor cell differentiation: Acceleration of in situ endothelialization
Authors: Royer Caroline , Guay‐Bégin Andrée‐Anne , Chanseau Christel , Chevallier Pascale , Bordenave Laurence , Laroche Gaétan , Durrieu Marie‐Christine
Published: March 2020

Synthetic grafts do not provide an appealing surface for endothelial cells to adhere and colonize the inner surface. To promote in situ endothelialization the following aspect has to be taken into account, endothelial progenitor cells (EPCs) needs to be mobilized on the surface of the graft. The surface of the graft has to be sufficiently biocompatible to create a prone environment for the EPCs to adhere, proliferate and, differentiate to form a layer and subsequently improve graft patency. In this work, two active molecules GRGDS and sitagliptin, were chosen for their abilities to recruit, enhance adhesion and induce differentiation of endothelial progenitor cells. They were grafted on PET surfaces in order to provide restrained cues triggering cell alignment and evaluate the influence of such structuration on EPCs fate. We then analyze cell behavior onto functionalized biomaterials. Their abilities to control EPCs fate were demonstrated via RT‐qPCR, immunofluorescence, and enzymatic tests. The GRGDS/sitagliptin 100 × 10 surface enables to reduce the stemness phenotype on EPCs and induce the expression of endothelial lineage markers. These results highlight the importance of spatial patterning cues in guiding EPCs organization and function, which may have clinical relevance in the development of vascular grafts that promote patency.
Biomaterials and biosensors in intestinal organoid culture, a progress review
Authors: Huang Jinjian , Jiang Yungang , Ren Yanhan , Liu Ye , Wu Xiuwen , Li Zongan , Ren Jianan
Published: March 2020

As an emerging technology, intestinal organoids are promising new tools for basic and translational research in gastroenterology. Currently, culture of intestinal organoids relies mostly on a type of tumor‐derived scaffolds, namely Matrigel, which may pose tumorigenic risks to organoid implantation. Apart from the traditional detection methods, such as tissue slicing and fluorescence staining, the monitoring of intestinal organoids requires real‐time biosensors that can adapt to their three‐dimensional dynamic growth patterns. In this review, we summarized the recent advances in developing definite hydrogel scaffolds for intestinal organoid culture and identified key parameters for scaffold design. In addition, classified by different substrate compositions like pH, electrolytes, and functional proteins, we concluded the existing live‐imaging biosensors and elucidated their underlying mechanisms. We hope this review enhances the understanding of intestinal organoid culture and provides more practical approaches to investigate them.
Review on calcium‐ and magnesium‐based silicates for bone tissue engineering applications
Authors: Venkatraman Senthil Kumar , Swamiappan Sasikumar
Published: March 2020

Bone is a self‐engineered structural component of the human body with multifaceted mechanical strength, which provides indomitable support to the effective functioning of the human body. It is indispensable to find a suitable biomaterial for substituting the bone as the bone substitute material requirement is very high due to the rate of bone fracture and infection lead to osteoporosis in human beings increases rapidly. It is not an easy task to design a material with good apatite deposition ability, a faster rate of dissolution, superior resorbability, high mechanical strength, and significant bactericidal activity. Since the synthetic hydroxyapatite was not able to achieve the dahlite phase of hydroxyapatite (natural bone mineral phase), silicates emerged as an alternate biomaterial to meet the need for bone graft substitutes. All silicates do not exhibit the properties required for bone graft substitutes, as their composition and methodology adopted for the synthesis are different. Calcium, magnesium, and silicon play a major role in the formation of bone mineral and their metabolism during bone formation. In this review, the relationship between composition and activity of calcium, magnesium‐based silicates have been discussed along with the future scope of these materials for hard tissue engineering applications.
Antibody‐modified magnetic nanoparticles as specific high‐efficient cell‐separation agents
Authors: Saei Arezoo , Asfia Shima , Kouchakzadeh Hasan , Rahmandoust Moones
Published: March 2020

Separation of tumor cells is a promising approach that helps not only in early detection of cancer but also as an efficient tool that holds great importance in prohibiting cancer cell mutation, drug resistance to treatments, and in granting successful adjuvant therapies. As one of the highly efficient processes for the separation of single cells, tumor cells, and specific proteins from fresh whole blood, a magnetic iron oxide nanoparticle (IONP)‐based immunomagnetic separation technique has been developed in this article. The synthesized IONPs were modified with antibodies (Abs) against human epithelial growth factor receptor 2 (HER2), which is overexpressed and/or amplified in about 15% of breast cancer patients with several types of human cancer cells. The prepared Ab‐conjugated IONPs (Ab‐IONPs) attach HER2‐positive cancer cells exclusively and can serve as specific high‐efficient single‐cell separation agents. The results showed that the magnetic IONPs have been successfully attached to the Abs via 1‐ethyl‐3‐(3‐dimethylaminopropyl) carbodiimide/N‐hydroxysuccinimide linkers. Maximum targeting efficiency of the Ab‐IONP complex, which was 94.5 ± 0.8% for BT474 and 70.6 ± 0.4% for mixture of cells (BT474 and MCF7), was achieved with a minimum amount of Abs, to provide an economically efficient single‐cell detection device.
Correlation of lipid peroxidation and nitric oxide metabolites, trace elements, and antioxidant enzymes in patients with sickle cell disease
Authors: Antwi‐Boasiako Charles , Dankwah Gifty Boatemaah , Aryee Robert , Hayfron‐Benjamin Charles , Aboagye George , Campbell Andrew D.
Published: March 2020

Background

Lipid peroxidation plays a very important role in sickle cell pathophysiology. The formation of malondialdehyde (MDA) in patients with sickle cell disease (SCD) may lead to endothelial dysfunction. Nitric oxide (NO) is a known vasodilator which plays a role in endothelial function. The current study determined the association between MDA and NO metabolites (NOx), trace elements, and antioxidant enzymes (SOD and CAT) in patients with SCD. The ratio of MDA/NOx was also determined as an index of oxidative stress in the study groups.

Methods

This was a cross‐sectional study involving 90 patients with SCD and 50 “healthy” controls. Blood samples (n = 140) were collected from the study groups. The plasma, sera, and red cells were kept at −20°C for biochemical analyses. Hemoglobin (Hb) and NOx levels were determined in the plasma using Labsystem Multiskan MS and Griess reagent system, respectively. Super oxide dismutase (SOD) and catalase (CAT) levels were determined in the red cells using assay kits from Cayman chemicals. Lipid peroxidation biomarker MDA was determined in the sera using the TBARS assay. Levels of iron (Fe), copper (Cu), and zinc (Zn) were also determined in the sera using Variant 240FS. MDA and NOx ratio was computed for the study groups and compared.

Results

Levels of Hb, NOx, SOD, CAT, and Zn were significantly lower in the patients with SCD (P < .001). MDA, Fe, and MDA/ NOx ratio were, however, significantly higher in the patients with SCD (P < .001). There was no significant correlation between MDA and NOx, SOD, CAT, Fe, and Zn in the study groups. MDA, however, correlated positively and significantly with Cu in the HbSS patients with vaso‐occlusive crises (VOC). Gender did not affect the levels of oxidative stress markers.

Conclusions

Findings from this study suggest a link between lipid peroxidation and Cu in HbSS patients with VOC. Increased MDA/NOx ratio may contribute to sickle cell pathophysiology by promoting oxidative stress.
Echocardiographic detection of left atrium and left ventricle tumoral invasion via the left upper pulmonary vein, leading to the diagnosis of lung cancer
Authors: Uygur Begum , Celik Omer , Birant Ali , Demir Ali R. , Erturk Mehmet
Published: March 2020

Left atrial (LA) invasion by lung cancer via hematogenous pathways is relatively uncommon. Herein we report the case of a 68‐year‐old male without any medical history, in whom lung cancer was diagnosed by transesophageal echocardiographic detection of the LA and left ventricle tumoral invasion via the left upper pulmonary vein. The primary source of tumor was found out by computed tomography.
Synthesis of chromenone, pyrimidinone, thiazoline, and quinolone derivatives as prospective antitumor agents
Authors: El‐Helw Eman A. E. , El‐Badawy Azza A.
Published: March 2020

Hydrazide‐hydrazone namely, 2‐cyano‐N′‐((1‐phenyl‐3‐[thiophen‐2‐yl]‐1H‐pyrazol‐4‐yl)methylene)acetohydrazide (3) underwent a series of reactions with some chemical reagents to construct new biologically active N‐heterocycles, for example, chromenone, benzochromenone, thiazoline, and quinolone derivatives. Treating the nitrile derivative 3 with 2,4‐dichlorobenzaldehyde and pyrazole aldehyde 1 afforded the corresponding condensed products. Some of the synthesized compounds were screened for their in vitro antitumor activities against two different human tumor cell lines including hepatocellular liver carcinoma (HepG2) and breast adenocarcinoma (MCF7) activities. Compound 3 was the most potent against the two tumors.
Harnessing neoantigen specific CD4 T cells for cancer immunotherapy
Authors: Brightman Spencer E. , Naradikian Martin S. , Miller Aaron M. , Schoenberger Stephen P.
Published: March 2020

The goal of precision immunotherapy is to direct a patient's T cell response against the immunogenic mutations expressed on their tumors. Most immunotherapy approaches to-date have focused on MHC class I-restricted peptide epitopes by which cytotoxic CD8+ T lymphocytes (CTL) can directly recognize tumor cells. This strategy largely overlooks the critical role of MHC class II-restricted CD4+ T cells as both positive regulators of CTL and other effector cell types, and as direct effectors of antitumor immunity. In this review, we will discuss the role of neoantigen specific CD4+ T cells in cancer immunotherapy and how existing treatment modalities may be leveraged to engage this important T cell subset.
Genotyping of enteroviruses and human parechoviruses highlights their diversity in Northwest Ethiopia
Authors: Gelaw Aschalew , Pietsch Corinna , Tigabu Zemene , Liebert Uwe G.
Published: March 2020

Enteroviruses (EVs) and human parechoviruses (HPeVs) infections are associated with various forms of disease, including gastroenteritis. As information on the molecular epidemiology of these viruses is limited in Ethiopia, the genetic diversity of EV and HPeV was investigated in the Northwestern part of the country. Of the total 450 stool samples obtained from infants and young children with diarrhea, 157 (34.9%) were positive for EV and 49 (10.9%) for HPeV RNA when tested by real‐time reverse transcription polymerase chain reaction. Genotyping was performed by sequencing of the EV VP1 gene and the HPeV VP3/VP1 gene, respectively. Genotyping of EV was successful in 118 samples. Thereof, 82 (69.5%) belonged to non‐polio EVs as a broad range of genotypes within species C, B, and A. Sabin polioviruses were found in 36 cases. HPeV sequences were also heterogeneous with a relative dominance of genotype 3. In conclusion, diverse EV and HPeV genotypes were found cocirculating in Northwest Ethiopia. The findings highlight the importance of continuous surveillance of these viruses in Ethiopia.
The application of in vitro‐derived human neurons in neurodegenerative disease modeling
Authors: D'Souza Gary X. , Rose Shannon E. , Knupp Allison , Nicholson Daniel A. , Keene Christopher Dirk , Young Jessica E.
Published: March 2020

The development of safe and effective treatments for age‐associated neurodegenerative disorders is an on‐going challenge faced by the scientific field. Key to the development of such therapies is the appropriate selection of modeling systems in which to investigate disease mechanisms and to test candidate interventions. There are unique challenges in the development of representative laboratory models of neurodegenerative diseases, including the complexity of the human brain, the cumulative and variable contributions of genetic and environmental factors over the course of a lifetime, inability to culture human primary neurons, and critical central nervous system differences between small animal models and humans. While traditional rodent models have advanced our understanding of neurodegenerative disease mechanisms, key divergences such as the species‐specific genetic background can limit the application of animal models in many cases. Here we review in vitro human neuronal systems that employ stem cell and reprogramming technology and their application to a range of neurodegenerative diseases. Specifically, we compare human‐induced pluripotent stem cell‐derived neurons to directly converted, or transdifferentiated, induced neurons, as both model systems can take advantage of patient‐derived human tissue to produce neurons in culture. We present recent technical developments using these two modeling systems, as well as current limitations to these systems, with the aim of advancing investigation of neuropathogenic mechanisms using these models.