bluuebunny/crossref_metadata_2025 huggingface dataset:
Abstract P-27: The 30S Ribosomal Subunit Assembly Factor Rbfa Plays a Key Role in the Formation of the Central Pseudoknot and in the Correct Docking of Helix 44 of the Decoding Center
Authors: Maksimova Elena , Korepanov Alexey , Kravchenko Olesya , Baymukhametov Timur , Myasnikov Alexander , Vassilenko Konstantin , Afonina Zhanna , Stolboushkina Elena
Published: October 2021

Background: Ribosome biogenesis is a complicated multi-stage process. In the cell, 30S ribosomal subunit assembly is fast and efficient, proceeding with the help of numerous assembly protein factors. The exact role of most assembly factors and mechanistic details of their operation remain unclear. The combination of genetic modification with cryo-EM analysis is widely used to identify the role of protein factors in assisting specific steps of the ribosome assembly process. The strain with knockout of a single assembly factor gene accumulates immature ribosomal particles which structural characterization reveals the information about the reactions catalyzed by the corresponding factor. Methods: We isolated the immature 30S subunits (pre-30S subunits) from the Escherichia coli strain lacking the rbfA gene (ΔrbfA) and characterized them by cryo-electron microscopy (cryo-EM). Results: Deletion of the assembly factor RbfA caused a substantial distortion of the structure of an important central pseudoknot which connects three major domains of 30S subunit and is necessary for ribosome stability. It was shown that the relative order of the assembly of the 3′ head domain and the docking of the functionally important helix 44 depends on the presence of RbfA. The formation of the central pseudoknot may promote stabilization of the head domain, likely through the RbfA-dependent maturation of the neck helix 28. The cryo-EM maps for pre-30S subunits were divided into the classes corresponding to consecutive assembly intermediates: from the particles with completely unresolved head domain and unfolded central pseudoknot to almost mature 30S subunits with well-resolved body, platform, and head domains and with partially distorted helix 44. Cryo-EM analysis of ΔrbfA 30S particles revealing the accumulation of two predominant classes of early and late intermediates (obtained at 2.7 Å resolutions) allowed us to suggest that RbfA participate in two stages of the 30S subunit assembly and is deeper involved in the maturation process than previously thought. Conclusion: In summary, RbfA acts at two distinctive 30S assembly stages: early formation of the central pseudoknot including the folding of the head, and positioning of helix 44 in the decoding center at a later stage. An update to the model of factor-dependent 30S maturation was proposed, suggesting that RfbA is involved in most of the subunit assembly process.
Abstract P-3: The Structure of Self-Assembled Surfactant Micellar Networks by in situ Cryo-Electron Tomography
Authors: Chesnokov Yury , Shibaev Andrey , Kamyshinsky Roman , Kralin Vyacheslav , Philippova Olga , Orekhov Anton
Published: October 2021

Background: Surfactant molecules can form various self-assembled structures in aqueous solutions, including spherical and cylindrical micelles, lamellae, vesicles, etc. Elongated cylindrical (wormlike) micelles can entangle and form a dense network. The study of the un-perturbed native structure of wormlike micelles in such networks presents a great challenge, since the micelles are formed due to weak non-covalent interactions and may easily break when external conditions are changed. In this work in situ cryo-electron tomography (cryo-ET) was applied to reveal the relaxed structure of such entangled systems. Methods: To prepare samples for the cryo-ET study 1 µl of the aqueous surfactant-containing solution was applied to the glow discharged grid, blotted with filter paper for 10 sec, drained for 60 sec to allow for the relaxation of the system and plunge-frozen with Vitrobot Mark IV. The vitrified sample was transferred to Versa 3D cryo-focused ion beam / scanning electron microscope (cryo-FIB/SEM) to prepare thin (100-150 nm) sections of the sample. Cryo-ET study was conducted using Titan Krios. IMOD and Avizo software packages were used for data processing. Results: In this work, wormlike micelles formed by a mixture of an anionic and a cationic surfactant were investigated at the excess of the anionic surfactant. Cryo-ET study of the obtained lamellae demonstrated the formation of two different phases, consisting of straight rods oriented along the grid substrate (phase 1) and isotropic network formed by wormlike micelles (phase 2) above it. The topology of the second phase corresponded to the branched saturated network or entangled network depending on cation/anion ratio of the sample. However, the analysis of the thin samples obtained without cryo-FIB demonstrated only the presence of the metastable phase (phase 1), which could lead to false conclusions regarding the morphology of the micelles. Conclusion: Here we discuss the influence of different sample preparation approaches on the sample structure and demonstrate that the native un-perturbed conformation of charged cylindrical surfactant micelles in the dense network is that of a slightly bent rod or a wormlike chain with high persistence length.
Abstract P-31: Assembly of the Complex of the 30S Ribosomal Subunit and the Ribosome Maturation Factor P from Staphylococcus aureus for Structural Studies by Cryo-Electron Microscopy
Authors: Garaeva Natalia , Bikmullin Aydar , Klochkova Evelina , Validov Shamil , Yusupov Marat , Usachev Konstantin
Published: October 2021

Background: Staphylococcus aureus (S. aureus) is one of the main human pathogens causing numerous nosocomial soft tissue infections and is among the best-known causes of bacterial infections. The bacterial 70S ribosome consists of two subunits, designated the 30S (small) and 50S (large) subunits. The small subunit (30S) consists of 16S ribosomal RNA (rRNA), from which the assembly of 30S begins, and 21 ribosomal proteins (r-proteins). The ribosome maturation factor P (RimP protein) binds to the free 30S subunit. Strains lacking RimP accumulate immature 16S rRNA, and fewer polysomes and an increased amount of unassociated 30S and 50S subunits compared to wild-type strains are observed in the ribosomal profile. Structural studies of the 30S subunit complex and the ribosome maturation factor RimP will make it possible in the future to develop an antibiotic that slows down or completely stops the translation of Staphylococcus aureus, which will complicate the synthesis and isolation of its pathogenic factors. Here we present the protocol of the in vitro reconstruction of S. aureus 30S ribosome subunit in a complex with RimP for further structural studies by cryo-electron microscopy. Methods: Recombinant RimP protein from S. aureus was expressed in E. coli and purified by Ni-NTA chromatography and size exclusion chromatography. Reconstitution of the 30S–RimP complex was performed by mixing RimP protein with 30S ribosome. Unbound RimP protein was removed by Amicon Ultra Concentration (Merk KGaA, Darmstadt, Germany) with a cut-off limit of 100 kDa. The presence of RimP protein in the resulting 30S-RimP complex was confirmed by SDS-PAGE, and the quality of the final sample was analyzed by the negative staining EM. Results: Finally, by in vitro reconstruction, the 30S-RimP complex from S. aureus was obtained for further structural studies by cryo-electron microscopy.
Abstract P-37: Phosphorous Mapping in Inactivated SARS-Cov-2 Particles by Electron Energy Loss Spectroscopy
Authors: Moiseenko Andrey , Kozlovskaya Lubov , Ishmukhametov Aydar , Egorov Alexey , Shaitan Konstantin , Kirpichnikov Mikhail , Sokolova Olga
Published: October 2021

Background: The severe COVID-19 pandemic started in December 2019 is caused by the SARS-CoV-2 virus. The SARS-CoV-2 virion consists of a positive-sense single-stranded RNA (ssRNA), bound with the nucleocapsid N protein and surrounded by a lipid membrane with the embedded glycoprotein S and the transmembrane proteins M and E. The structure of inactivated SARS-CoV-2 virions is crucial for the development of vaccine-induced immunity. Here we characterized the nucleic acid distribution within β-propiolactone inactivated whole-virion SARS-CoV-2 vaccine CoviVac. Methods: We used EELS to verify the presence of phosphorus (P) inside the β-propiolactone inactivated virions. Electron microscopy was performed with a JEM-2100 200kV LaB6 transmission electron microscope (JEOL, Japan) equipped with a Gatan GIF Quantum ER energy filter (Gatan, USA) operating in spectrometer mode, along with a High-Angle Annular Dark-Field (HAADF) scanning transmission electron microscopy (STEM) detector. The cooling holder model 21090 (JEOL, Japan) was operated at -182 °С to reduce the contamination effects and to enhance the specimen's stability under the electron beam. We employ a negative stain with 2% (NH4)2MoO4 rather than uranyl acetate since the Uranium O4,5 peak (edge at 96 eV) is close to the P L2,3 peak (edge at 132 eV) and interferes with the accurate background interpolation. Results: The intensity under the P peak after the background subtraction was used for STEM-EELS mapping. We observed the characteristic P signal from the inner part of the virion but not from the bare grid. The observed P signal could arise from either viral RNA or lipids of the virus membrane, and since the P signal is highly heterogeneous, it is more likely to originate from RNA. Conclusion: So far, phosphorous mapping in individual virions using EELS was done only with samples prepared using highly specialized techniques, which minimized the sample thickness, including the substrate thickness. Here, we performed elemental mapping on ordinary samples of whole viruses. All investigated virions contained P signal, but its spatial distribution and intensity differed significantly. This clearly reflects the non-even distribution of the genomic RNA, which, apparently, accompanies their inner heterogeneity, previously observed by in-situ cryo-electron tomography.
Abstract P-38: Tunable Soft Networks of Wormlike Micelles and Clay Particles
Authors: Molchanov Vyacheslav , Orekhov Anton , Arkharova Natalia , Kuklin Alexandr , Rogachev Andrei , Philippova Olga
Published: October 2021

Background: Over the past few decades, there has been a great deal of interest in the aqueous self-assembly of surfactant molecules into giant wormlike micelles (WLMs). These cylindrical aggregates undergo reversible breakdown processes and in favorable cases can grow up to few tens of micrometers that is comparable with the length of high molecular weight polymer. The viscoelastic properties of WLMs can be easily modified by different additives like salts or polymers. A new emerging research area consists of tuning the WLM solution properties by inorganic nanoparticles. It suggests, in particular, the use of networks of entangled WLMs as a matrix for producing soft nanocomposites with different kinds of embedded nanoparticles that are promising for controlled release, template synthesis, and oilfield applications. These materials can combine adaptive rheological properties of the WLM matrix and the functionality of nanofiller. Methods: Rheometry and cryo-transmission electron microscopy were combined to investigate the structure and properties of mixed WLMs of zwitterionic oleylamidopropyl dimethyl betaine and anionic sodium dodecyl sulfate surfactants and platelike particles of bentonite clay. Results: This system demonstrates the formation of giant linear long-lived WLMs, which even at extremely low surfactant concentrations reach a sufficient length to entangle with each other and form a temporally persistent network. The stability of these micelles can be due to electrostatic attraction between the headgroups of the anionic and zwitterionic surfactants and favorable volume/length hydrophobic ratio in the surfactant mixture. At increasing surfactant concentration, the long-lived linear micelles transform into fast-breaking branched micelles. Stable viscoelastic suspensions of clay particles in semi-dilute solutions of WLM were elaborated. They represent a novel type of soft nanocomposite with the tunable matrix. Structural studies revealed that the clay is dispersed in a dense network of entangled WLM in the form of 100-nm tactoids. Rheological investigations demonstrated that clay particles can induce an increase of viscosity and relaxation time by up to one order of magnitude. The effect of the clay becomes more pronounced with increasing content of anionic surfactant, when the micelles become branched. This behavior was explained by the stabilization of micelle-nanoclay junction points due to the screening of the repulsion between positively charged fragments of zwitterionic head groups by added anionic surfactant. Conclusion: The pronounced effect of nanoparticles on the viscoelasticity of the network formed by branched WLMs was observed for the first time. The nanoparticles-WLM junctions were confirmed by cryo-TEM data. The elaborated systems are of interest for many industrial applications.
Abstract P-46: Structure of A. Baumannii Phage Tapaz, Revealed with Cryo-Electron Microscopy
Authors: Moiseenko Andrey , Wang Yueqi , Litvinov Daniil , Popova Anastasia , Shneider Mikhail , Miroshnikov Konstantin , Sokolova Olga
Published: October 2021

Background: Acinetobacter baumannii is an opportunistic pathogen and one of the six most important multidrug resistant microorganisms in hospitals worldwide. Some of its strains are resistant to most of the antibiotics, A. baumannii is included into the Priority 1 part of Global Priority List of Antibiotic-resistant Bacteria. Phage therapy is considered to be an alternative strategy to antibiotic treatments. Methods: A. baumannii strain NIPH601 cells were grown till OD6000.4 and infected with the phage at MOI 10:1. After complete lysis took place cell debris was spined down and phage particles were precipitated with the PEG6000 (final concentration 10% PEG 6000, 0.5 NaCl). Virus particles were collected by centrifugation, resuspended at SM buffer and applied on CsCl step gradient. Gradient was spinned down for 2 hours at 40000g and the fraction containing phage particles was collected and dialyzed against SM buffer. Purified phage particles were applied to Quantifoil 1.2/1.3 grids and plunge-froze in Vitrobot Mark IV (TFS) Micrographs were collected in HKU, Shenzhen campus with Titan Krios cryoelectron microscope (TFS), equipped with Gatan K3 direct electron detector. The micrographs were acquired with 1.06 Å pixel size and 1.5 um average defocus value in counting mode with 50 frames and 1.2 e/Å2/frame dose rate. All image processing was performed with Relion3.0 software, except for the particle picking step performed with cryolo. Results: Lytic A. baumannii phage TaPaz belongs to the family Myoviridae. BLAST search over NCBI “nr” (non-redundant) database revealed close homology with previously published sequences of Acinetobacter phage vB_AbaM_B9 and Acinetobacter phage BS46. However, no structural information about any homologous proteins was found among the PDB structures. The cryo-EM map was reconstructed with single particle analysis independently for the capsid, tail and baseplate regions. The capsid was reconstructed at 3.9 Å resolution with I3 symmetry applied (Fig. 1A). The baseplate region of the phage was reconstructed at 3.5 Å resolution with C3 symmetry (Fig. 1B). The tail region was reconstructed at 2.6 Å resolution with helical symmetry (Rise 36.4 Å, Twist 25.7 deg). Initial atomic model for the tail region was built from sequence with Deeptracer and was further refined in coot (Fig. 1C). Conclusion: We successfully obtained the near-atomic resolution structural map of phage TaPaz. The data obtained contribute to enhancing knowledge of structural diversity of bacterial viruses infecting A. baumannii.
Abstract P-6: Automated Pipeline for Parametrization of the Coarse-Grained Models for Biomolecular Simulations
Authors: Orekhov Philipp
Published: October 2021

Background: Antibiotic-resistant strains of Staphylococcus aureus cause human infections that are difficult to treat and can lead to death. Bacteriophage (phage) phi812K1/420 from the family Myoviridae infects 95% of clinical isolates of S. aureus and therefore is a promising candidate for a phage therapy agent. As the native phage particle approaches its host cell, phage receptor-binding proteins make a contact with the host cell wall. This interaction triggers a cascade of structural changes in the baseplate resulting in phage tail contraction and genome ejection. Mechanistic description of the baseplate re-organization, however, remains unknown. Methods: Using cryo-electron microscopy (cryo-EM), we studied the baseplate of the phage phi812K1/420. Also, selected proteins involved in the host cell wall binding and penetration were produced in recombinant form and their structures were solved using X-ray crystallography and cryo-EM single-particle reconstruction. Results: We reconstructed the phage baseplate in native and contracted states. The reconstruction of the native baseplate reaches a resolution of 4 Å, which enables us to discern individual protein structures. Solved protein structures will be fitted into the reconstruction of the contracted baseplate. Conclusion: Our results provide the first structural characterization of contractile phage infecting a Gram-positive bacterium. Comparison of the two distinct baseplate states will allow us to describe the molecular mechanism of the initial stage of phage infection in detail.
Análise comparativa da eficiência alocativa das despesas públicas municipais utilizando mineração de dados
Authors: Guelman Davis Paula , De Souza Antônio Artur
Published: October 2021

Objetivo da pesquisa: Analisar, comparativamente, a eficiência alocativa das despesas públicas dos municípios brasileiros utilizando técnicas de mineração de dados. Entende-se, como eficiente, a melhor relação entre o volume de despesas e resultados nos indicadores socioeconômicos, ou seja, menores despesas e melhores resultados quando comparado aos outros municípios.

Enquadramento teórico: Teorias da Federação, especificamente o Federalismo Fiscal, na qual uma das principais proposições é a de que tributos e despesas publicas descentralizadas são mais eficientes.

Metodologia: Coleta e a integração de dados financeiros e operacionais. Transformação em indicadores. Limpeza e seleção de indicadores para transformação em atributos categóricos (discretização). Geração de árvores de decisão, a partir de algoritmos de mineração de dados, permitindo identificar características em comum dos municípios com desempenho destacado.

Resultados: Viabilidade e importância do uso da mineração de dados sobre conjuntos abrangentes de dados. Foram identificados padrões relevantes nas áreas de educação, saúde e desenvolvimento, como a importância de se despender recursos na formação dos docentes, e caracterizadas discrepâncias regionais em termos do desempenho do Sistema Único de Saúde (SUS).

Originalidade: Nos estudos anteriores, foi possível constatar uma repetição da metodologia Data Envelopment Analysis (DEA) para estudo da eficiência das despesas públicas. Em contraste com esses estudos, utilizamos grandes conjuntos de dados municipais, de múltiplas fontes e áreas, aplicando técnicas de mineração de dados para encontrar padrões relevantes.

Contribuições teóricas e práticas: Proposição de uma metodologia de estudo da eficiência da despesa pública municipal, válida do ponto de vista teórico e prático, em múltiplas áreas de atuação, com as técnicas e ferramentas da Mineração de Dados.
Investimento em educação diminui a criminalidade: análise dos municípios do Rio Grande do Sul usando econometria espacial
Authors: Fochezatto Adelar , Petry Guilherme , Braatz Jacó , Marcondes Henrique Romão
Published: October 2021

A criminalidade tem sido um dos principais problemas da sociedade brasileira. No Brasil, percebe-se uma concentração de crimes na faixa etária entre 15 e 19 anos, que coincide com a idade escolar, principalmente de ensino médio. Vários estudos defendem que investimentos na primeira infância para promover a aprendizagem são fundamentais, pois é neste período da vida que a pessoa desenvolve suas habilidades e as noções de moralidade e civilidade. Esta aprendizagem acumula capital humano e, ao longo do ciclo de vida, promove melhores oportunidades, diminuindo a propensão à criminalidade. Além disso, a simples retenção do aluno na escola e o seu envolvimento com os estudos podem diminuir as chances dos jovens se envolverem com a contravenção. O acesso, a retenção e o rendimento do aluno na escola dependem de investimentos em infraestrutura e em recursos humanos na área da educação. Este estudo procura identificar se há associação entre gastos em educação e taxas de criminalidade nos municípios do Rio Grande do Sul. Para isto é usado um modelo simples de econometria espacial, que permite verificar também efeitos de transbordamentos entre municípios. Os resultados obtidos permitem afirmar que um aumento dos gastos públicos em educação tende a diminuir a criminalidade.
EDUCATIONAL WARGAMING: Design and Implementation into Professional Military Education
Authors: Combe II P. C.
Published: October 2021

In light of the Commandant’s Planning Guidance, there is a renewed emphasis on educational wargaming in professional military education (PME). While wargaming has a long history in PME, there is currently a gap in the academic literature regarding wargaming as an adult educational tool. Scientific study has focused on adult education theory and models generally, highlighting the identification of four different learning experiences, each tied to a learning style: concrete experience, which suits those with a diverging earning style; abstract conceptualization, which suits those with the converging learning style; reflective observation, for those with an assimilating learning style; and active experimentation, which works well for those with an accommodating learning style. By effectively engaging each of these four experiences, educational wargaming can have utility for a diverse array of learning styles.
ASSESSMENT STRATEGIES FOR EDUCATIONAL WARGAMES
Authors: Kuehn Kate
Published: October 2021

Purposeful integration of assessment within educational wargame design is increasingly essential as military education expands those activities within its curriculum. This multimethod case study examines key challenges and strategies for assessment within educational wargaming practice. Drawing insights from faculty interviews, academic documents, and faculty meeting observations, the study identifies six key assessment challenges: gamesmanship, lack of control, multiple faculty roles, receptiveness to feedback, evaluation of individuals in teams, and fairness of evaluation. It then discusses how experienced faculty mitigate these challenges throughout the assessment design process from identifying outcomes to ensuring the quality of evaluation.
WARGAMING DEVELOPMENT SERIES: Developing Impactful Wargame Narratives through Storytelling
Authors: Gordon Stephen M. , Yates Walt , Gordon Andrew
Published: October 2021

Nothing connects people more powerfully than well-told stories. Humans have been telling each other stories since long before they could write them down. Sharing stories is a critical part of building trust with others, and that trust is essential to creating meaningful connections with people. Great stories have structure and purpose; they appeal to our deepest emotions and are most compelling when they challenge or change our perceptions of reality. There are rules to the methods and techniques that create great stories. This article explores the benefits and challenges of applying successful storytelling techniques to designing wargame narratives that balance creative ambitions with achievable timelines. Wargames that incorporate such techniques will surface new trends and better inform future conflict planning.
ENZYMATIC AND AFLATOXIN PRODUCTION POTENTIAL OF ASPERGILLUS FLAVUS
Authors: Sana Saba
Published: October 2021

Fungi especially Aspergillus species are potential candidates for production of mycotoxins and industrially important enzymes. Aspergillus flavus isolates (129) recovered from soil mixed with animal rations (n=145) had aflatoxins (17.82%) and Enzymes (10.37%) production potential. Quantity of detected Aflatoxins varied for different isolates i.e., 3.25 to 11622.24ng, 21.34 to 194.47ng and 3.36 to 40.12ng per mL of Sabouraud’s dextrose broth in case of AFB1, AFB2 and AFG1 as determined by High performance liquid chromatography. Optimization of non-toxigenic starch hydrolyzing A. flavus was carried out at different incubation temperatures (22, 30 and 37°C), pH (4.5, 6 and 7.5) and substrates including maize, wheat bran and rice husk (1, 3 and 5% each) for incubation period of 7 days. In optimization experiments for starch hydrolysis, most of the A. flavus (86%) produced highest enzyme (IU) at 37°C and pH 6 quantified by Dinitrosalicylic method. Most of the isolates were able to produce enzymes using rice husk followed by maize. The highest quantity of enzyme was produced by A. flavus (179.88+1.71IU) using one percent of maize at pH 6 and 37°C. It was concluded that indigenous non-toxigenic A. flavus can be used in food industry as biological source of starch hydrolyzing enzymes.
BIOCHEMICAL AND HISTOPATHOLOGICAL INVESTIGATIONS OF ANTIDIABETIC POTENTIAL OF POLYHERBAL FORMULATION IN ALLOXAN INDUCED DIABETIC RATS
Authors: Iftikhar Asra
Published: October 2021

Herbal therapies always remain a potential source of glycemic control in diabetic patients. Plants or combination of plants improve the healing action and lessen the adverse effects compared to synthetic drugs. The present research was designed to evaluate antidiabetic potential of polyherbal formulation consisting of Sphaeranthus indicus, Caesalpinia bonduc, Bunium persicum, Artemisia absinthium, Cuminum cyminum, Swertia chirata, Gymnema sylvestra, and Citrullus colocynthis in alloxanized diabetic rats. Quantitative phytochemical, RP-HPLC, DPPH and in vitro alpha amylase analyses of formulation were performed. Rats were grouped into six groups (n=15/group) named as control, diabetic control, glibenclamide treated, Treated I, Treated II and Treated III. Hyperglycemic rats of treated groups I, II and III were administered polyherbal formulation at dose rate of 200, 400 and 600mg/kg respectively for 8 weeks after induction of diabetes with alloxan monohydrate (150, IP). The results have shown that alloxan significantly increased fasting blood glucose (531.81±4.29) and serum glucose (526.0±6.26) levels. However, 8-weeks treatment of hyperglycemic rats with polyherbal formulation significantly decreased the fasting blood glucose (156.82±4.51), serum glucose (148.8±4.04) and increased C-peptide and liver glycogen levels. The levels of ALT, AST, ALP, TAC and TOS were also restored towards normal. Histopathological analysis of pancreas also showed increase in size and number of pancreatic β cells after polyherbal formulation treatment compared to diabetic group. This study shows that polyherbal formulation has antidiabetic potential as it can improve performance of pancreatic β cells and insulin secretory capacity by reducing hyperglycemia and oxidative stress induced β cells apoptosis
IMPROVING THE PRODUCTIVITY AND SUGAR RECOVERY OF CANE BY POTASH NUTRITION UNDER DIFFERENT PLANTING METHODS
Authors: Aslam Cheema Muhammad Waqas
Published: October 2021

Potassium-nutrition is a subject of great consideration, as research has revealed its importance in increasing sugar recovery in sugarcane. While it cannot achieve its genetic expression of yield without proper planting technology. Therefore, the present study was conducted to optimize the planting method and K level to improve the yield and sugar recovery of sugarcane at University of Agriculture Faisalabad in 2014-2016. Treatments comprised of four planting methods viz. i) 90 cm spaced pits with a diameter of 90 cm, ii) 90 cm spaced pits with a diameter of 90 cm in diagonal fashion, iii) 90 cm spaced double row strips and iv) 120 cm spaced trench planting in combination with four K nutrition levels i.e., 0, 100, 200 and 300 kg/ha K2O. Results revealed that K application improved the growth, cane yield and sugar recovery irrespective of planting method. However, maximum number of tillers were recorded in sugarcane sown in diagonal pit planting at 90 cm with 100 kg/ha K2O. Likewise, Leaf area index and net assimilation rate were substantially improved with K application in all planting methods. More cane weight (15%) was obtained at sugarcane sown in 120 cm spaced trenches with 200 kg/ha K2O during both the years. Stripped cane yield was maximum in diagonal pit planting at 90 cm with 100 kg/ha K2O (113.7 t/ha) during the plant crop year and diagonal pit plantation + 200 kg/ha K2O (98.22 t/ha) during the ratoon crop year. Likewise, maximum potassium use efficiency (KUE) i.e. 99.5 and 88.2 kg /kg were recorded in planting of sugarcane in diagonal pits at 90 cm with 100 kg/ha K2O during plant crop year and 120 cm trenches +100 kg/ha K2O during the ratoon crop year, respectively. Sugar recovery was also enhanced by all the K nutrition levels over control. The cultivation of sugarcane in 90 cm spaced pits with the supplementation of potash at 100 kg/ha gave the maximum sugar yield of 15.8 t/ha in plant crop year and 13.2 t/ha in the ratoon crop year. The combined economic analysis over two years (plant + ratoon year) revealed that sugarcane planting was more beneficial at 120 cm spaced trenches, with 100 kg potash/ha (3678$); which was followed by 90 cm diagonal pit plantation +100 kg/ha K2O which gave the combined benefits of 3611$. Sugar cane may be planted in 90 cm diagonal pits and 120 cm spaced trenches with 100 kg/ha potash to improve the cane yield and sugar recovery
EFFECT OF PHOSPHORIC ACID AND POTASSIUM HUMATE ON GROWTH AND YIELD OF MAIZE IN SALINE-SODIC SOIL
Authors: Ameen Muhammad
Published: October 2021

Phosphorus (P) is the 2nd most deficient nutrient among plants (<10 mg P kg−1 soil) in >90% soil of Pakistan. The crop requirement is achieved by fertilization however effectiveness of the current P (Phosphorus) containing fertilizers applied as broadcast is less (20%). Application of phosphoric acid (PA) and potassium humate (PH) not only increases the nutrients availability to plant but also improves soil characteristics. A pot experiment was carried out to evaluate the behavior of PA and PH in saline-sodic soil for nutrient availability in maize. Twenty-seven plastic pots were used in this experiment. PA was applied according to recommended dose of Punjab Agriculture Department (NPK 200-150-200 kg ha-1 ), while the PH was applied according to the Agrolix fertilizer private limited recommendation (50 and 100 kg ha-1 ). The plant growth parameters (length and dry weight of root and shoot), ionic interaction (N, P, K, Fe, Zn, Cu and Mn in root, shoot and grain) and grain weight were recorded. The results showed 241% and 315% more shoot and root dry weight as compared to control with combined application of PA and PH. The length of shoot and root were improved to 181% and 232% as compared to control. The grain yield was positively correlated with shoot dry weight (r2 = 0.6618). Application of PA decreased the soil electrical conductivity (EC) and sodium adsorption ratio (SAR) about 91% and 96% respectively after one cropping season. Grain yield had highly significant (r2= 0.829) negative correlation with soil ECe (Electrical conductivity) whereas it was also negatively correlated with soil SAR (r2=0.5942). Results further explained the significant positive correlation between phosphorus and iron concentration in grain (r2 = 0.7454). However, phosphorus concentration was not correlated with grain zinc concentration (r2 = 0.1798).
Implementing Specialized Vestibular Physiotherapy in An Emergency Department: A Process Evaluation
Authors: Ip Kelvin , Lloyd Melanie , Luscombe Allison , Hitch Danielle
Published: October 2021

Background: Dizziness and vertigo-like symptoms, often caused by common peripheral vestibular disorders such as Benign Paroxysmal Positional Vertigo (BPPV), may have a significant detrimental impact on function and quality of life. The impact of these symptoms often result in Emergency Department (ED) presentations. Evidence based clinical practice guidelines strongly recommend the use of physical assessment and treatment maneuvers for the assessment, diagnosis and treatment of these symptoms. The aim of this study was to evaluate the process of implementing specialized vestibular physiotherapy in an emergency department, from the clinician perspective.Methods: This implementation study utilized a retrospective mixed methods process evaluation to understand how specialized vestibular physiotherapy operated in an Australian emergency department. The PARiHS Framework was embedded within the methodology and analytical approach of the study, to ensure a comprehensive approach which was closely aligned to implementation science. Nine clinicians retrospectively completed the Organizational Readiness for Change Assessment (ORCA), Acceptability of Intervention Measure (AIM), Intervention Appropriateness Measure (IAM) and Feasibility of Intervention Measure (FIM). Seven clinicians also participate in a focus group or interview.Results: A range of barriers and facilitators to the implementation process were identified by participants, some of which spanned multiple domains of the PARiHS framework. Relationships with service leaders, champions and medical staff were found to be a key facilitator to implementation, along with a generally held perception that specialized vestibular physiotherapy was acceptable and feasible. The main barrier identified was a lack of capacity to deliver and support this innovation, both within the physiotherapy workforce and the broader multidisciplinary team.Conclusions: This study demonstrates the process of implementation of a specialized vestibular physiotherapy team in an ED setting was generally well received by clinicians, but also involved some challenges and barriers. Services looking to implement specialized vestibular physiotherapy in the ED may refer to the recommendations arising from the findings of this study to guide their approach to innovation.
Intact Predictive Motor Sequence Learning in Autism Spectrum Disorder
Authors: Rybicki Alicia J. , Galea J. M. , Schuster Bianca , Hiles C. , Fabian C. , Cook J. L.
Published: October 2021

Background. Atypical motor learning has been suggested to underpin the development of motoric challenges (e.g., handwriting difficulties) in autism. Bayesian accounts of autistic cognition propose a mechanistic explanation for differences in the learning process in autism. Specifically, that autistic individuals overweight incoming, at the expense of prior, information and are thus less likely to a) build stable expectations of upcoming events and b) react to statistically surprising events. Although Bayesian accounts have been suggested to explain differences in learning across a range of domains, to date, such accounts have not been extended to motor learning.Methods. 28 autistic and 35 non-autistic controls (IQ > 70) completed a computerised task in which they learned sequences of actions. On occasional “surprising” trials, an expected action had to be replaced with an unexpected action. Sequence learning was indexed as the reaction time difference between blocks which featured a predictable sequence and those that did not. Surprise-related slowing was indexed as the reaction time difference between surprising and unsurprising trials.Results. No differences in sequence-learning or surprise-related slowing were observed between the groups. Bayesian statistics provided anecdotal to moderate evidence to support the conclusion that sequence learning and surprise-related slowing were comparable between the two groups. Conclusions. We conclude that individuals with autism do not show atypicalities in response to surprising events in the context of motor sequence-learning. These data demand careful consideration of the way in which Bayesian accounts of autism can (and cannot) be extended to the domain of motor learning.
Classification of Hypertensive and Normotensive Subjects Using Bilateral Differential Biopotential Signals
Authors: Sarkar Arindam , Goswami Bhaswati , Ghosh Ratna
Published: October 2021

Hypertension or high blood pressure is a severe health issue in the modern world, especially in this pandemic scenario, that can cause many heart related diseases or even death, and it is increasing day by day. For this reason, a
reliable, automatic and easy to use system for hypertensive subject detection is an important focus for the researchers. Biopotential signals can play a pivotal role in this regard. Though, few strategies were proposed based on electrocardiogram (ECG) or electrodermal (EDA) signals, but those require special circuitry, as well as trained persons. In this article, a method is proposed to classify hypertensive and normotensive subjects using differential biopotential
signals. Neither special circuitry, nor much expertise is required for handling this system. It was assumed that progression of rest is dependent upon blood pressure. To serve the purpose, signals were acquired from both hypertensive and normotensive subjects bilaterally for 10 continuous minutes. Result of the random forest (RF) classification establishes that from the analysis of the progression of the bilaterally acquired differential biopotential signals, hypertensive subjects can be distinguished from normotensive subjects.
Optimization of an Economical Medium Composition For The Coculture of Clostridium Butyricum And Bacillus Coagulans
Authors: Li Yonghong , Wang Yun , Liu Yingying , Li Xuan , Li Keke , Feng Lifei
Published: October 2021

Clostridium butyricum is a spore-forming probiotic existing in the intestines of humans and animals which can promote the enhancement of beneficial bacteria and maintain intestinal microecological balance. However, it is difficult to improve the production level of C. butyricum by conventional fermentation process. In this study, a co-fermentation process of C. butyricum DL-1 and Bacillus coagulans ZC2-1 was established to improve the viable counts and spore yield of C. butyricum, and the formula of coculture medium was optimized by flask fermentation. The results showed that the optimum medium composition is bran 10 g/L, corn steep powder 15 g/L, peptone 15 g/L, K2HPO4 1 g/L and MnSO4 0.5 g/L. Cultured stationarily in the optimal medium for 36 h, the number of viable bacteria of C. butyricum DL-1 reached 1.5×108 CFU/mL and the spore forming rate was 92.6%. The results revealed an economical and effective medium composition for the coculture of C. butyricum and B. coagulans. The co-fermentation process established in this study provides a new fermentation mode for the industrial production of other absolute anerobic bacteria.
The effectiveness of oral immunotherapy in patients with Sesame anaphylaxis using Omalizumab
Authors: Salari Fereshteh , Bemanian Mohammad Hassan , Fallahpour Morteza , Mahdaviani Seyed Alireza , Shokri Sima , Khoshmirsafa Majid , Seif Farhad , Nabavi Mohammad , Arshi Saba
Published: October 2021

Objective

Sesame allergy is the most prevalent allergy to seeds. Oral immunotherapy (OIT) is defined as continuous consumption of an allergen at special doses and time. Omalizumab (Anti-IgE) increases tolerance to allergens used in OIT. This study evaluated the effectiveness of a new sesame OIT protocol in patients with sesame anaphylaxis in combination with omalizumab.
Methods

In this prospective open-label interventional trial study, 11 patients with a history of sesame anaphylaxis were enrolled after approval by Oral Food Challenge (OFC) test. At baseline, skin prick test (SPT) and skin prick to prick (SPP) test were performed. Serum sesame-specific IgE (sIgE) levels were measured. The maintenance phase was continued at home with daily sesame intake for 4 months. At the end of month 4, the OFC and above-mentioned tests were repeated to evaluate the treatment effectiveness.
Results

All 11 patients who underwent sesame OIT after 4 months could tolerate a dietary challenge of 22 ml tahini (natural sesame seed, equal to 5,000 mg of sesame protein and higher) and the average of wheal diameter in the SPT and SPP tests significantly decreased after desensitization.
Conclusion

This OIT protocol may be a promising desensitization strategy for patients with sesame anaphylaxis. Also, omalizumab obviously reduced the severity of reactions.
Robot-assisted Anterior Odontoid Screw for the Treatment of Type Ⅱ Odontoid Fractures: Safety and Effectiveness Analysis
Authors: Zhao Songchuan , Bo Yang , Du Jinpeng , Yan Liang , Hao Dingjun , Wang Haosheng , Wei Jianmin , Gao Lin
Published: October 2021

Background: Anterior odontoid screw fixation is considered to be preferred surgical treatment for the type Ⅱ odontoid fractures. However, due to the high difficulty to insert odontoid screw with barehand, the high risk of screw misalignment and damage to surrounding important tissue structures, we urgently need robot-assisted screw insert navigation technology to improve the safety and accuracy of inserting odontoid screws.Methods: We retrospectively analyzed 7 patients with type II odontoid fractures who underwent Tinavi robot-assisted screw insert technology from May 2018 to May 2019 at our hospital. All patients had received 64-row CT scans and 3D reconstructions completed preoperatively, and magnetic resonance (MRI) were performed to verify the severity of odontoid fractures, soft tissue injuries and vertebral artery height. Postoperative CT was repeated in 6 months after surgery to evaluate cervical stability and confirm whether the screw had breached the bone cortex, the accuracy of screw placement based on Rampersaud A-D grade. Functional recovery was assessed using the post-traumatic Mayor scoring system for the cervical spine.Results: All 7 patients successfully completed the robot-assisted operation without nerve and blood vessel damage. What is the operation time 103.3 minutes, intraoperative blood loss 11.1 ml. The angulation and displacement of the fracture were basically corrected by closed reduction during the operation. Postoperative CT of these 7 patients showed that the cervical spine was stable, the accuracy of “perfect” and “clinically acceptable” odontoid screw implantation was 100% (7/7), none of the seven odontoid screws breached the bone cortex. Reexamination of X-rays showed that the fractures were all healed, and the average fracture healing time was average 13.7weeks (12-15weeks). During the follow-up period, 7 patients had no surgical complications, postoperative cervical spine trauma mayo score: excellent in 6 cases and good in 1 case. Conclusion: Tinavi robot-assisted screw insert technology is a minimally invasive, accurate, safe and feasible method for the treatment of type Ⅱ odontoid fractures.
FOXA3: A Novel Tumor Suppressor in Esophageal Squamous Cell Carcinoma
Authors: Zhang Siang , Jin Yuxiang , Han Qianyu , Zhao Xuewei , Xue Lei
Published: October 2021

Background: Forkhead box A(FOXA) family, have been widely investigated in cancer research. However, currently the study of FOXA3 in malignant tumors is still limited compared with other family members. FOXA1 and FOXA2 both play a role in the development and progression of esophageal cancer and Barrett's esophagus, but the exact role of FOXA3 in esophageal squamous cell carcinoma (ESCC) has not been elucidated.Methods: The expression of FOXA3 in ESCC tissues was determined by real-time PCR immunohistochemistry staining. The relationship between FOXA3 and overall survival rate of patients was analyzed by Kaplan-Meier analysis, and Cox regression analysis was used to analyze independent prognostic risk factors. Cell proliferation was analyzed by CCK-8 assay, EdU assay and colony forming assay. Cell migration was analyzed by wound healing assay and Transwell assay. Tumorigenesis by subcutaneous injection was performed to detect the inhibition of tumors by FOXA3 in vivo.Results: We found that FOXA3 is down-regulated in ESCC tissues and low FOXA3 expression significantly correlated with a poor prognosis of ESCC. Additionally, after up-regulating the FOXA3 expression, the abilities of proliferation, migration and invasion in ESCC cells were significantly inhibited. Moreover, over-expression of FOXA3 can inhibit tumor growth in vivo either via tumorigenesis assay by subcutaneous injection.Conclusions: Low FOXA3 expression significantly correlated with a poor prognosis of ESCC and the over-expression of FOXA3 could inhibit the growth and metastasis of ESCC, which indicated that FOXA3 maybe a novel biomarker and therapeutic target for ESCC.
WITHDRAWN: Identification of Novel Biomarkers and Targets for Natural Products in Inhibiting Prostate Cancer
Authors: Li Wenji , Xu Wei , Sun Kai , Wang Fujun , Wong Tin Wui
Published: October 2021

Background: Prostate cancer (PCa) is a common urinary system malignancy. The lack of specific and sensitive biomarkers for the diagnosis and prognosis of PCa makes it important to seek alternatives. Meanwhile, targeted PCa inhibitors are limited. Natural products that potentially target PCa may offer a useful approach.Methods: Expression profile datasets about PCa from GEO were analyzed. Core differential genes were identified by String and Cytoscape. GEPIA and HPA were utilized to further validate the key genes. The targets of natural products were obtained from the Drugbank, Therapeutic Target Database, BindingDB, PubChem, and chEMBL databases, and PCa therapeutic targets were generated from the GeneCards, OMIM, and PharmGkb databases. Cytoscape was also used to screen the core modules and disease-drug targets. Construction of molecular docking models of drug-core targets was performed by Autodock to confirm the accuracy of the targets.Results: Four identified biomarkers, CENPF, TPX2, TK1 and CCNB1 were verified by HPA. Five novel PCa biomarkers, RRM2, UBE2C, TOP2A, BIRC5 and ZWINT were also identified. All the nine markers indicated poor prognosis for PCa patients were verified by GEPIA. PCa carcinogenesis is found to be mainly associated with hepatic fibrosis pathway, ILK signaling, NRF2-mediated oxidative stress response and many others. Four key PCa targets for curcumin (EP300, RELA, EGFR, NFKB1), seven for taxol (PTEN, EGFR, ERBB2, TP53, KRAS, AR, AKT1) and two for ursolic acid (GSK3B, RELA) were identified by Cytoscape combined KEGG and verified by Autodock.Conclusions: The novel identified biomarkers in our study would be valuable for the diagnosis and prognosis of PCa. Key targets of curcumin, paclitaxel, and ursolic acid in PCa could lay a solid foundation for precise treatment and molecularly targeted therapy for PCa.