We report a facile route to the first molecular compounds with the Al-O-M-O-Al (M=Ti, Zr) structural motif. Synthesis of L(Me)Al(mu-O)M(NMe2)2(mu-O)Al(Me)L [L=CH{N(Ar)(CMe)}2, Ar=2,6-iPr2C6H3; M=Ti (7), Zr (8)] was accomplished by reacting the monometallic hydroxide precursor L(Me)Al(OH) (1) with Ti(NMe2)4 or Zr(NMe2)4 under elimination of Me2NH in good yield. The crystal structural data confirm the trimetallic Al-O-M-O-Al core in both 7 and 8. Preliminary investigation on catalytic activity of these complexes reveals low activity of these complexes in ethylene polymerization as compared to the related oxygen-bridged metallocene-based heterobimetallic complexes L(Me)Al(mu-O)M(Me)Cp2 (M=Ti, Zr) which could be attributed to the relatively lower stability of the supposed cationic intermediate as revealed by DFT calculations.

Two single oxygen-bridged heterobimetallic oxides of Al(III) with group 4 metals (Ti, Hf) have been prepared. The reaction of LAlMeOH (1) [L = CH(N(Ar)(CMe))2, Ar = 2,6-iPr2C6H3] with dimethylmetallocenes of Ti and Hf in toluene (80 degrees C) and ether (room temperature), respectively, resulted in the formation of LAl(Me)(mu-O)M(Me)Cp2 [M = Ti (2), Hf (3)] in moderate to good yield. Compounds 2 and 3 were characterized by elemental analysis, IR, NMR (1H and 13C), EI-MS, and single-crystal X-ray structural analysis. Furthermore, compound 2 showed good catalytic activity in ethylene and styrene homopolymerization, while compound 3 is less active in ethylene polymerization. The styrene polymerization yields atactic polystyrene.

Partial abstraction of apical methyl groups is observed in aminopyridinatoniobium alkyne complexes, as exemplified with the synthesis and characterization of 1. This results in cationic complex fragments, which facilitate the polymerization of ethylene and in the presence of methylalumoxane the selective dimerization of 1-butene.

Background/aim: Myofibroblastoma of the breast is a rare benign mesenchymal tumor whose morphology is similar to that of spindle-cell lipoma. The few hitherto genetically investigated mammary myofibroblastomas have been shown to have had loss of material from chromosome 13, changes that are also common in spindle-cell lipoma. Our aim was to add to the existing knowledge of genetic aberrations in mammary myofibroblastoma by investigating another such tumor.

Materials and methods: Cytogenetic and array comparative genome hybridization (aCGH) analyses were performed on a surgically removed mammary myofibroblastoma from a 76-year-old man.

Results: Short-term cultured cells from the tumor showed the karyotype 45,XY,-13[3]/44~45,idem,add(19)(q13)[cp2]. aCGH detected loss of one entire chromosome 13 and heterozygous loss from 19q between sub-band 19q13.12 and 19qter.

Conclusion: These findings add to the evidence that loss of 13q material is typical of mammary myofibroblastomas.

Keywords: Mammary myofibroblastoma; chromosome 13; cytogenetics; deletion.

Ferroptosis is an iron- and lipid peroxidation-dependent form of regulated cell death. The release of labile iron is one of the important factors affecting sensitivity to ferroptosis. Yes-associated protein (YAP) controls intracellular iron levels by affecting the transcription of ferritin heavy chain (FTH) and transferrin receptor (TFRC). However, whether YAP regulates iron metabolism through other target genes remains unknown. Here, we observed that the system Xc^- inhibitor erastin inhibited the binding of the WW domain and PSY motif between YAP and transcription factor CP2 (TFCP2), and then suppressed the transcription of ferritin light chain (FTL) simultaneously mediated by YAP, TFCP2 and forkhead box A1 (FOXA1). Furthermore, inhibition of FTL expression abrogated ferroptosis-resistance in cells with sustained YAP expression. Unlike FTH, which exhibited first an increase and then a decrease in transcription, FTL transcription continued to decline after the addition of erastin, and a decrease in lysine acetyltransferase 5 (KAT5)-dependent acetylation of FTL was also observed. In lung adenocarcinoma (LUAD) tissues, lipid peroxidation and labile iron decreased, while YAP, TFCP2 and FTL increased compared to their adjacent normal tissues, and the lipid peroxidation marker 4-hydroxynonenal (4-HNE) was negatively correlated with the level of FTL or the degree of LUAD malignancy, but LUAD tissues with lower levels of 4-HNE showed a higher sensitivity to ferroptosis. In conclusion, the findings from this study indicated that the suppression of FTL transcription through the inhibition of the YAP-TFCP2-KAT5 complex could be another mechanism for elevating ferroptosis sensitivity and inducing cell death, and ferroptotic therapy is more likely to achieve better results in LUAD patients with a lower degree of lipid peroxidation.

Keywords: LUAD; TFCP2; YAP; erastin; labile iron; lipid peroxidation.

The CP2 transcription factors are highly conserved in metazoans, where they are divided into two groups: grainyhead and late SV40 factor (LSF). We traced their evolutionary history in the Hexapoda using over 500 insect transcriptomes, to test the hypothesis that the evolution of holometaboly involved novel isoforms of these genes. All insects appear to express at least one grainyhead and one LSFlike gene, regardless of life cycle, as in most known metazoa. No major evolutionary events in these gene families occurred during the evolution of insects.

The overuse of chloramphenicol (CAP) due to its low price is detrimental to ecological safety and human health. An earthworm gut content dwelling bacterium, Aeromonas media SZW3, was isolated with capability of CAP biodegradation, and the CAP degradation efficiency reached 55.86% at day 1 and 67.28% at day 6. CAP biodegradation kinetics and characteristic of strain SZW3 determined the factors that affect CAP biodegradation. Thirteen possible biodegradation products were identified, including three novel biodegradation products (CP1, CP2 and CP3), and three potential biodegradation pathway were proposed. Biodegradation reactions include amide bond hydrolysis, nitro group reduction, acetylation, aminoacetylation, dechlorination and oxidation. Genome analysis suggested that the coding gene of RarD (CAP resistance permease), CAP O-acetyltransferase, nitroreductase and haloalkane dehalogenase may be responsible for CAP biodegradation. The proposed complete biodegradation pathway and genome analysis by strain SZW3 provide us new insight of the transformation route and fate of CAP in the environment.

Keywords: Aeromonas media SZW3; Biodegradation pathway, Genome analysis; Chloramphenicol (CAP) biodegradation.

Rice tungro disease (RTD) is a recurring disease affecting rice farming especially in the South and Southeast Asia. The disease is commonly diagnosed by visual observation of the symptoms on diseased plants in paddy fields and by polymerase chain reaction (PCR). However, visual observation is unreliable and PCR can be costly. High-throughput as well as relatively cheap detection methods are important for RTD management for screening large number of samples. Due to this, detection by serological assays such as immunoblotting assays and enzyme-linked immunosorbent assay are preferred. However, these serological assays are limited by lack of continuous supply of antibodies as reagents due to the difficulty in preparing sufficient purified virions as antigens. This study aimed to generate and evaluate the reactivity of the recombinant coat proteins of Rice tungro bacilliform virus (RTBV) and Rice tungro spherical virus (RTSV) as alternative antigens to generate antibodies. The genes encoding the coat proteins of both viruses, RTBV (CP), and RTSV (CP1, CP2 and CP3) were cloned and expressed as recombinant fusion proteins in Escherichia coli. All of the recombinant fusion proteins, with the exception of the recombinant fusion protein of the CP2 of RTSV, were reactive against our in-house anti-tungro rabbit serum. In conclusion, our study showed the potential use of the recombinant fusion coat proteins of the tungro viruses as alternative antigens for production of antibodies for diagnostic purposes.

DH (Doubled haploid) is the immortal mapping population and an outcome of single meiotic cycle, contributed from male partner. An improved procedure was developed for high frequency androgenesis in japonica genotypes, K-332 and GS-88 and their F1s. A total of 207 fertile, green, di-haploid plants were generated from K-332 × GS-88 hybrids using the improved anther culture protocol. The investigation was carried out to evaluate callus induction potential and regeneration response for the genotypes and the derived F1s on N6 media and modified N6 media (N6M). Whereas, N6 failed to induce callusing, agarose solidified N6M media supplemented with 4% maltose, growth regulators; NAA (2 mg/l), 2, 4-D (0.5 mg/l), Kinetin (0.5 mg/l), and silver nitrate induced high calli percentage of 27.6% in F1s, 9.5% and 6.7% in GS-88 and K-332 respectively. Murashige and Skoog (MS) media supplemented with 3% sucrose, and the hormonal combination BAP (2 mg/l), Kinetin (1 mg/l) and NAA (1 mg/l) induced high green shoot regeneration rates (0-60.0%). The effect of cold pre-treatment at 4°C and the stage of anther collection and their interaction was studied. The effect of cold pre-treatment (CP) of collected boots at 4°C (for CP2: 2, CP4: 4, CP6: 6 and CP8: 8 days) at different stages of panicle emergence (BES4-6: 4-6, BES7-10: 7-10, BES11-13: 11-13, BES>13: more than 13 inches was worked out in relation to the effect on response of calli induction, albino regeneration, green plant regeneration and number of shoots/green calli. CP referred to the number of days for which the collected boots were incubated before they were inoculated. BES was the length (inches) between flag leaf and penultimate leaf at the time of boot collection. We concluded that CP6 and BES7-10 showed better response to callus proliferation and regeneration of plantlets across genotypes. The appropriate pre-treatment, stage of anther collection and favourable media composition resulted in high calli induction and green plant regeneration rates in recalcitrant japonica genotypes. The modified N6 media resulted into efficient callus induction and is expected to be useful for studies which aim at rapid generation of mapping populations for genetic studies.

Pathology and putative virulence factor expression of three Histomonas meleagridis isolates differing in geographic origin, cell passage number (56 or 100), or cell populations grown from a monoculture were compared. Turkey poults inoculated with the high cell passage number isolates or monoculture isolates varied in gross lesion severity and weight gain (P<0.0001). Screening of a published H. meleagridis cDNA library identified forty- eight cysteine proteinases (CP) and one superoxide dismutase (Fe-SOD) proposed to function in either tissue damage and/or invasion and oxidative defense. The Fe-SOD and eight CPs were analyzed using real time polymerase chain reaction. CP2, CP3, and CP8 showed significant differences in expression among the field isolates (P ≤ 0.05). The high passage isolates had decreased CP2, CP3 and CP4 expression when compared with their field isolate. CP7 did not differ between field isolates or the 56-passaged isolate. The Fe-SOD gene showed significant differences in expression among the various isolates. When exposing cultured H. meleagridis to air, Fe-SOD expression decreased rapidly during the first hour of air exposure but increased progressively through the next 3 h. This study provides information on gross pathology and virulence factors associated with various isolates of Histomonas meleagridis which can aid in determining the pathogenetic mechanisms used by this organism.

Keywords: Blackhead disease; Entamoeba histolytica; Histomonasmeleagridis; Trichomonas vaginalis; Turkeys.

(Arginine-alanine-aspartic acid-alanine)₄ ((RADA)₄) nanoscaffolds are excellent candidates for use as peptide delivery vehicles: they are relatively easy to synthesize with custom bio-functionality, and assemble in situ to allow a focal point of release. This enables (RADA)₄ to be utilized in multiple release strategies by embedding a variety of bioactive molecules in an all-in-one "construct". One novel strategy focuses on the local, on-demand release of peptides triggered via proteolysis of tethered peptide sequences. However, the spatial-temporal morphology of self-assembling nanoscaffolds may greatly influence the ability of enzymes to both diffuse into as well as actively cleave substrates. Fine structure and its impact on the overall effect on peptide release is poorly understood. In addition, fractal networks observed in nanoscaffolds are linked to the fractal nature of diffusion in these systems. Therefore, matrix morphology and fractal dimension of virgin (RADA)₄ and mixtures of (RADA)₄ and matrix metalloproteinase 2 (MMP-2) cleavable substrate modified (RADA)₄ were characterized over time. Sites of high (glycine-proline-glutamine-glycine+isoleucine-alanine-serine-glutamine (GPQG+IASQ), CP1) and low (glycine-proline-glutamine-glycine+proline-alanine-glycine-glutamine (GPQG+PAGQ), CP2) cleavage activity were chosen. Fine structure was visualized using transmission electron microscopy. After 2 h of incubation, nanofiber networks showed an established fractal nature; however, nanofibers continued to bundle in all cases as incubation times increased. It was observed that despite extensive nanofiber bundling after 24 h of incubation time, the CP1 and CP2 nanoscaffolds were susceptible to MMP-2 cleavage. The properties of these engineered nanoscaffolds characterized herein illustrate that they are an excellent candidate as an enzymatically initiated peptide delivery platform.

Regions required for chicken glycine decarboxylase gene transcription were examined. A region between -82 and +22 (-82/+22) with motifs similar to binding sites for Sp1, NF-Y and CP2 was assigned to the proximal promoter active in both chicken hepatoma cell line, LMH, and hepatocytes in primary culture. In LMH cells, a genomic region, KX, between KpnI (-4155) and XbaI (-2113) sites changed promoter activity with the aid of four additional genomic regions termed upstream regulator regions for suppression (UpRS) and activation (UpRA) of transcription. Those precise segments are UpR1S (-376/-346), UpR1A (-345/-291), UpR2S (-137/-108) and UpR2A (-107/-83). Within KX, -4155/-3605 activates and -3604/-3367 suppresses the promoter. -3366/-3024 activates or suppresses the promoter, probably with different UpR counterparts. -2197/-2113 restores the actions of -3366/-3024. While in LMH cells, the upstream UpRs abrogate the functions of immediately downstream UpRs, UpR1S or UpR2S or both may be at least less active in hepatocytes than in LMH cells. Nuclear extracts from various chicken tissues and LMH cells had UpR2A binding proteins in different populations, suggesting that together with the UpRs, the segments in KX are involved in the regulation of cell type-specific transcription of this gene.

Cysteine/cystine redox couple plays a vital role in maintaining the cellular redox state of living bio-systems. However, the dynamic changes of the intracellular cysteine/cystine pool mediated by cysteine/glutamate transporter (System X_C^-) in various cellular processes have remained incompletely understood. Here we established a series of strategies around fluorescent probe CP2 for effectively monitoring the intracellular cysteine level in living cells and in vivo zebrafish model, therefore for tracking dynamic changes of system X_C^--mediated cysteine/cystine pool under ferroptosis. Living cell imaging and in vivo visualization both revealed that intracellular cysteine levels could be significantly reduced upon blocking system X_C^- activity and inducing ferroptotic cell death. This kind of blocking was potent against cisplatin-resistant A549R cancer cells, which might bring new potential for treating drug-resistant carcinoma through regulating system X_C^- and processes of ferroptosis. With further improvements in monitoring tools, understanding of system X_C^--mediated cystine/cysteine pool and ferroptosis could certainly offer crucial information in more physiological or pathological processes.

Keywords: Cysteine/glutamate transporter (system X(C)(−)); Ferroptosis; Fluorescent imaging strategy; Intracellular cysteine/cystine pool; Zebrafish.

Stemness and epithelial-mesenchymal transition (EMT) are two fundamental characteristics of metastasis that are controlled by diverse regulatory factors, including transcription factors. Compared with other subtypes of breast cancer, basal-type or triple-negative breast cancer (TNBC) have high frequencies of tumor relapse. However, the role of alpha-globin transcription factor CP2 (TFCP2) has not been reported as an oncogenic driver in those breast cancers. Here we show that TFCP2 is a potent factor essential for EMT, stemness, and metastasis in breast cancer. TFCP2 directly bound promoters of epidermal growth factor (EGF) and transforming growth factor alpha (TGF-α) to regulate their expression and stimulate autocrine signaling via epidermal growth factor receptor (EGFR). These findings indicate that TFCP2 is a new anti-metastatic target and reveal a novel regulatory mechanism in which a positive feedback loop comprising EGF/TGF-α and AKT can control malignant breast cancer progression.

Purpose: The aim of the study was to compare the lingualized implant placement creating a buccal cantilever with prosthetic-driven implant placement exhibiting excessive crown-to-implant ratio.

Materials and methods: Based on patient's CT scan data, two finite element models were created. Both models were composed of the severely resorbed posterior mandible with first premolar and second molar and missing second premolar and first molar, a two-unit prosthesis supported by two implants. The differences were in implants position and crown-to-implant ratio; lingualized implants creating lingually overcontoured prosthesis (Model CP2) and prosthetic-driven implants creatingan excessive crown-to-implant ratio (Model PD2). A screw preload of 466.4 N and a buccal occlusal load of 262 N were applied. The contacts between the implant components were set to a frictional contact with a friction coefficient of 0.3. The maximum von Mises stress and strain and maximum equivalent plastic strain were analyzed and compared, as well as volumes of the materials under specified stress and strain ranges.

Results: The results revealed that the highest maximum von Mises stress in each model was 1091 MPa for CP2 and 1085 MPa for PD2. In the cortical bone, CP2 showed a lower peak stress and a similar peak strain. Besides, volume calculation confirmed that CP2 presented lower volumes undergoing stress and strain. The stresses in implant components were slightly lower in value in PD2. However, CP2 exhibited a noticeably higher plastic strain.

Conclusion: Prosthetic-driven implant placement might biomechanically be more advantageous than bone quantity-based implant placement that creates a buccal cantilever.

Keywords: Bone resorption; Dental prosthesis, Implant-supported; Dental stress analysis; Finite element analysis.

Members of the Trichodoridae can cause substantial crop losses directly by feeding on plant roots and indirectly as vectors of tobraviruses; both vector and virus are polyphagous. Although trichodorid nematodes are important pests of agricultural crops, no data are available on the presence or extent of these nematodes in the Czech Republic. In June 2005, three soil samples from the rhizosphere of a Quercus sp. at Cerveny Vrch yielded a population of Trichodorus similis Seinhorst, 1963. Specimens were extracted from soil by a decanting-sieving method, heat killed, and fixed in triethanolamine formalin (TAF), and processed and mounted in anhydrous glycerin. Nematodes were identified by morphological and morphometrical characters (2). Classical identification of these nematodes was further verified by molecular study. A single, male specimen was temporarily mounted in distilled water on a glass slide and relaxed with gentle heat. Measurements and photographs were taken, and the specimen was transferred to a 0.5-ml Eppendorf tube containing 0.25 M NaOH. Total genomic DNA was prepared by a rapid technique (4). Morphometric data of the male specimen used for DNA study are: body length 867 μm; body width 81 μm; onchiostyle length 44 μm; spicule length 36 μm; distance of anterior cervical papilla (CP)1 from anterior end 39 μm, CP1 to CP2 25 μm, CP2 to CP3 22 μm; posterior precloacal supplement (SP1) to cloacal opening 27 μm, distance SP1 to SP2 32 μm, distance SP2 to SP3 39 μm. The following primers were used in the PCR reaction: species-specific sense primer SIMIREV2 (5'-CACTCGTCGGACTCAAACC-3') and universal antisense primer UNIVERSAL (5'-CCCGTCGCTACTACCGATT-3') (1). A single fragment of approximately 452 bp was amplified. The D2 and D3 expansion regions of the large subunit 28S rDNA were amplified using the primer D2A (5'-ACAAGTACCGTGAGGGAAAGTTG-3') and D3B (5'-TCGGAAGGAACCAGCTACTA-3' (3). The region was sequenced after purification of PCR products from the gel slice with a Qiagen gel purification kit (Qiagen Inc., Valencia, CA). The obtained sequence was deposited in Genbank (Accession No. DQ832183). The obtained sequence was compared by BLAST in NCBI and the results showed strong similarities with T. similis (Accession No. AM180730). To our knowledge, this is the first report of T. similis associated with a deciduous forest in the Czech Republic. Taking into account the agricultural importance of trichodorids and tobraviruses as plant pathogens, there is a need for a comprehensive survey of these taxa in the Czech Republic. The damage level threshold is in the case of virus vector species equivalent to a single nematode. Therefore, information on these plant parasites would be useful for developing nematode management strategies. References: (1) K. Boutsika et al. Plant Pathol. 53:110, 2004. (2) W. Decraemer and P. Baujard. Fundam. Appl. Nematol. 21:37, 1998. (3) P. De Ley et al. Nematology 2:591, 1999. (4) J. M. Stanton. Australas. Plant Pathol. 27:112, 1998.

Background: The SARS-CoV-2 pandemic has had profound implications on healthcare institutions.

Aims: This study aims to assess and compare referral patterns during COVID-19 to corresponding dates for the preceding 3 years (2017-2019), in order to preemptively coordinate the logistics of the surgical unit for similar future experiences.

Methods: Retrospective review for our institution, a national tertiary referral centre for spine pathology. Two distinct time-points were chosen to represent the varied levels of social restriction during the current pandemic: (i) study period 1 (SP1) from 11 November 2020 to 08 June 2020 represents a national lockdown, and (ii) study period 2 (SP2) from 09 June 2020 to 09 September 2020 indicates an easing of restrictions. Both periods were compared to corresponding dates (CP1: 11 March-08 June and CP2 09 June-09 September) for the preceding 3 years (2017-2019). Data collected included age, gender, and mechanism of injury (MOI) for descriptive analyses. MOIs were categorised into disc disease, cyclist, road-traffic-accident (RTA), falls < 2 m, falls > 2 m, malignancy, sporting injuries, and miscellaneous.

Results: All MOI categories witnessed a reduction in referral numbers during SP1: disc disease (-29%), cyclist (-5%), RTAs (-66%), falls < 2 m (-39%), falls > 2 m (-17%), malignancy (-33%), sporting injuries (-100%), and miscellaneous (-58%). Four of 8 categories (RTAs, falls < 2 m, malignancy, miscellaneous) showed a trend towards return of pre-lockdown values during SP2. Two categories (disc disease, falls > 2 m) showed a further reduction (-34%, -27%) during SP2. One category (sporting injuries) portrayed a complete return to normal values during SP2 while a notable increase in cyclist-related referrals was witnessed (+ 63%) when compared with corresponding dates of previous years.

Conclusion: Spinal injury continues to occur across almost all categories, albeit at considerably reduced numbers. RTAs and falls remained the most common MOI. Awareness needs to be drawn to the reduction of malignancy-related referrals to dissuade people with such symptoms from avoiding presentation to hospital over periods of social restrictions.

Keywords: COVID-19; Coronavirus; Referrals; SARS-CoV-2; Spinal injury; Spine surgery.

Identifying how plant species diversity varies across environmental gradients remains a controversial topic in plant community ecology because of complex interactions among putative factors. This is especially true for grasslands where habitat loss has limited opportunities for systematic study across broad spatial scales. Here we overcome these limitations by examining restored plant community responses to a large-scale precipitation gradient under two common Conservation Reserve Program (CRP) restoration approaches. The two restoration strategies examined were <em>CP2</em>, which seeds a relatively low number of species, and <em>CP2</em>5, which seeds a higher number of species. We sampled plant communities on 55 CRP fields distributed along a broad precipitation gradient (410-1170 mm mean annual precipitation) spanning 650 km within the grassland biome of North America. Mean annual precipitation (MAP) was the most important predicator of plant species richness which had a positive, linear response across the gradient. To a lesser degree, restoration practices also played a role in determining community diversity. The linear increase in species richness across the precipitation gradient reflects the species pool increase from short to tallgrass prairie communities and explained most of the richness variation. These findings provide insight into the diversity constraints and fundamental drivers of change across a large-scale gradient representing a wide variety of grassland habitats. Across a broad environmental gradient, initial planting differences between restoration practices had lower effects on plant diversity than expected. This suggests that new strategies are needed to effectively establish diverse plant communities on large-scale, restorations such as these.

Keywords: Conservation Reserve Program (CRP); Kansas (USA); conservation practices (CPs); environmental gradients; plant community ecology; plant diversity constraints; productivity-diversity relationship; restored grasslands; species pool hypothesis.

Circular RNAs (circRNAs) are correlated with the occurrence and progression of differentiated thyroid cancer (THCA). However, the regulatory mechanism of circRNAs in differentiated THCA is unclear. In the present study, we analyzed the circRNA microarray dataset (GSE93522) of thyroid tumors and discovered that circRNA HACE1 (circHACE1) was downregulated in differentiated THCA. We detected circHACE1 expression by quantitative real-time polymerase chain reaction (qRT-PCR). Gain-of-function experiments were performed to analyze the biological function of circHACE1 in differentiated THCA cells in vitro. The regulatory mechanism of circHACE1 in differentiated THCA was explored through bioinformatics analysis, dual-luciferase reporter, RIP (RNA immunoprecipitation), and/or RNA pull-down assays. The biological function of circHACE1 in THCA was confirmed by xenograft assay. We verified that circHACE1 was downregulated in differentiated THCA. Also, differentiated THCA patients with low circHACE1 expression were associated with TNM grade, lymphoid node metastasis, tumor size, and poor prognosis. CircHACE1 overexpression decreased xenograft tumor growth in vivo and induced cell cycle arrest, apoptosis, impeded proliferation, migration, and invasion in differentiated THCA cells in vitro. CircHACE1 could function as a microRNA (miR)-346 sponge and regulated Tfcp2L1 (transcription factor CP2 like 1) expression. MiR-346 overexpression offset circHACE1 elevation-mediated effects on malignant behaviors of differentiated THCA cells. Furthermore, Tfcp2L1 silencing counteracted the suppressive impact of miR-346 inhibitor on the malignancy of differentiated THCA cells. In conclusion, circHACE1 adsorbed miR-346 and elevated Tfcp2L1 expression, thus curbing cell malignancy in differentiated THCA, manifesting that circHACE1 might be a target for differentiated THCA treatment.

Keywords: Thyroid cancer; Transcription factor CP2 like 1; circHACE1; miR-346.

Thorough knowledge of the germination behavior of weed species could aid in the development of effective weed control practices, especially when glyphosate resistance is involved. A study was conducted using two glyphosate-resistant (GR) (SGW2 and CP2) and two glyphosate-susceptible (GS) (Ch and SGM2) populations of Chloris virgata, an emerging and troublesome weed species of Australian farming systems, to evaluate their germination response to different alternating temperature (15/5, 25/15 and 35/25°C with 12 h/12 h light/dark photoperiod) and moisture stress regimes (0, -0.1, -0.2, -0.4, -0.8 and -1.6 MPa). These temperature regimes represent temperatures occurring throughout the year in the eastern grain region of Australia. Seeds germinated in all the temperature regimes with no clear indication of optimum thermal conditions for the GR and GS populations. All populations exhibited considerable germination at the lowest alternating temperature regime 15/5°C (61%, 87%, 49%, and 47% for Ch, SGM2, SGW2, and CP2, respectively), demonstrating the ability of C. virgata to germinate in winter months despite being a summer annual. Seed germination of all populations was inhibited at -0.8 and -1.6 MPa osmotic potential at two alternating temperature regimes (15/5 and 35/25°C); however, some seeds germinated at 25/15°C at -0.8 MPa osmotic potential, indicating the ability of C. virgata to germinate in arid regions and drought conditions. Three biological parameters (T10: incubation period required to reach 10% germination; T50: incubation period required to reach 50% germination; and T90: incubation period required to reach 90% germination) suggested late water imbibition with increasing moisture stress levels. The GR population SGW2 exhibited a distinctive pattern in T10, T50, and T90, possessing delayed germination behaviour and thus demonstrating an escape mechanism against pre-plating weed management practices. Knowledge gained from this study will help in developing site-specific and multi-tactic weed control protocols.

We investigated the use of citrus pulp (CP) as a novel prebiotic capable of exerting microbiota and immunomodulating capacities to alleviate weaning stress. Inulin (IN), a well-known prebiotic, was used for comparison. Hundred and 28 male weaned piglets of 21 days old were assigned to 32 pens of 4 piglets each. Piglets were assigned to one of the four treatments, i.e., control, IN supplemented at 0.2% (IN0.2%), and CP supplemented either at 0.2% (CP0.2%) or at 2% (CP2%). On d10-11 and d31-32 post-weaning, one pig per pen was euthanized for intestinal sampling to evaluate the growth performance, chyme characteristics, small intestinal morphology, colonic inflammatory response and barrier integrity, metabolite profiles [gas chromatography-mass spectrometry (GC-MS), and liquid chromatography-mass spectrometry (LC-MS)], and microbial populations. The IN treatment and the two CP treatments induced higher small intestinal villus height to crypt depth ratios in comparison with the control diet at both sampling times. All treatments decreased acidic goblet cell absolute counts in the crypts in comparison to the control diet of the duodenum on d10-11 and d31-32. The gene expression of β-defensin 2 was downregulated in colonic tissues following the IN and CP2% inclusion on d31-32. On d31-32, piglets fed with IN and CP0.2% showed lower mRNA levels of occludin and claudin-3, respectively. Not surprisingly, flavonoids were observed in the colon in the CP treatments. Increased colonic acetate proportions on d10-11, at the expense of branched-chain fatty acid (BCFA) levels, were observed following the CP2% supplementation compared to the control diet, inferring a reduction of proteolytic fermentation in the hindgut. The beneficial microbial community Faecalibacterium spp. was promoted in the colon of piglets fed with CP2% on d10-11 (p = 0.04; false discovery rate (FDR) non-significant) and on d31-32 (p = 0.03; FDR non-significant) in comparison with the control diet. Additionally, on d31-32, CP2% increased the relative abundance of Megasphaera spp. compared to control values (p = 0.03; FDR non-significant). In conclusion, CP2% promoted the growth of beneficial bacterial communities in both post-weaning time points, modulating colonic fermentation patterns in the colon. The effects of CP supplementation were similar to those of IN and showed the potential as a beneficial feed supplement to alleviate weaning stress.

Keywords: barrier function; citrus pulp; gut morphology; inflammation; intestinal health; inulin; metabolites; microbiota.

Additive manufacturing enabled the development of personalized, ideally fitting medical devices. The topography of the surface of the 3D-printed implant may not only facilitate its integration but also cause its rejection, as the surface may become a reservoir for different bacterial strains. In this study, the innovative, raw, 3D- printed fracture fixation plates, manufactured by using selective laser melting (SLM) from Ti-6Al-4V were compared with commercially available, surface-modified plates commonly used in orthopedic surgery. The topography surface of the plates was studied by atomic force microscopy. Susceptibility to the development of biofilm was tested for Staphylococcus epidermidis, Staphylococcus aureus and Streptococcus mutans by using crystal violet staining of biomass, confocal, and scanning electron microscopy (SEM). 3D- printed plates showed higher roughness (Sa=131.0 nm) than commercial plates (CP1 and CP2), Sa= 60.67 nm and Sa=55.48 nm, respectively. All strains of bacteria colonized 3D- printed raw plates more densely than commercial plates. The microscopic visualization showed biofilm mostly in irregular cavities of printed plates while on commercial plates it was mainly located along the edges. The research has indicated that there is need for further development of this technology to optimize its effectiveness and safety.

Keywords: 3D printing; Biofilm; Biomaterials; Fracture fixation devices; Implant infections; Orthopedic implants; Patient-specific implants.

This study investigated attentional bias toward game-related cues in Internet gaming disorder (IGD) using electrophysiological markers of late positive potential (LPP) and identifying the sources of LPP. In addition, the association between LPP and decision-making ability was investigated. The IGD (n = 40) and healthy control (HC; n = 39) participants viewed a series of game-related and neutral pictures, while their event-related potentials (ERPs) were recorded. LPPs were calculated as the mean amplitudes between 400 and 700 ms at the centro-parietal (CP3, CP1, Cpz, CP2, and CP4) and parietal (P3, P1, Pz, P2, and P4) electrode sites. The source activations of LPP were estimated using standardized low-resolution brain electromagnetic tomography (sLORETA). In addition, decision-making ability was evaluated by the Cambridge Gambling Task. Higher LPP amplitudes were found for game-related cues in the IGD group than in the HC group. sLORETA showed that the IGD group was more active in the superior and middle temporal gyri, which are involved in social perception, than in the HC group, whereas it was less active in the frontal area. Individuals with IGD have deficits in decision-making ability. In addition, in the HC group, the lower the LPP when looking at the game-related stimuli, the better the quality of decision-making, but not in the IGD group. Enhanced LPP amplitudes are associated with emotional arousal to gaming cues and decision-making deficits in IGD. In addition, source activities suggest that patients with IGD perceive game-related cues as social stimuli. LPP can be used as a neurophysiological marker of IGD.

Interacting with objects in our environment usually leads to audible noise. Brain responses to such self-initiated sounds have been shown to be attenuated, in particular the so-called N1 component measured with electroencephalography (EEG). This attenuation has been proposed to be the effect of an internal forward model that allows for cancellation of the sensory consequences of a motor command. In the current study we asked whether the attenuation due to self-initiation of a sound also affects a later event-related potential - the so-called motion-onset response - that arises in response to moving sounds. To this end, volunteers were instructed to move their index fingers either left or rightward which resulted in virtual movement of a sound either to the left or to the right. In Experiment 1, sound motion was induced with in-ear head-phones by shifting interaural time and intensity differences and thus shifting the intracranial sound image. We compared the motion-onset responses under two conditions: a) congruent, and b) incongruent. In the congruent condition, the sound image moved in the direction of the finger movement, while in the incongruent condition sound motion was in the opposite direction of the finger movement. Clear motion-onset responses with a negative cN1 component peaking at about 160 ms and a positive cP2 component peaking at about 230 ms after motion-onset were obtained for both the congruent and incongruent conditions. However, the motion-onset responses did not significantly differ between congruent and incongruent conditions in amplitude or latency. In Experiment 2, in which sounds were presented with loudspeakers, we observed attenuation for self-induced versus externally triggered sound motion-onset, but again, there was no difference between congruent and incongruent conditions. In sum, these two experiments suggest that the motion-onset response measured by EEG can be attenuated for self-generated sounds. However, our result did not indicate that this attenuation depended on congruency of action and sound motion direction.

Keywords: Auditory evoked potentials; Efference copy; Electroencephalography; Motion-onset response; Sensorimotor integration; Sound motion.