Miracle-dz/UniFAR-FTUs huggingface dataset:
Mutations in Domain I of Bacillus thuringiensis -Endotoxin CryIAb Reduce the Irreversible Binding of Toxin to Manduca sexta Brush Border Membrane Vesicles (*)

Site-directed mutagenesis was used to generate CryIAb mutants at the selected N-terminal positions to study the function of domain I. Structurally stable mutant proteins were tested for toxicity, receptor binding kinetics, and pore function. Substitutions of tyrosine at position 153 with arginine (Y153R) or alanine (Y153A) did not affect toxicity appreciably, whereas replacing this tyrosine with aspartic acid (Y153D) resulted in a great loss of toxicity. Mutation of alanine at position 92 to glutamic acid (A92E) almost completely abolished toxicity. The initial receptor binding was unchanged as measured by competition binding assays among all mutant proteins. Reduced pore function, however, was observed for mutants A92E and Y153D as tested by voltage clamping. Further studies with specially designed association and dissociation binding assays showed that irreversible binding of these two mutant toxins to Manduca sexta brush border membrane vesicles was significantly reduced. The decrease in irreversible binding was correlated with the changes in toxicity and may reflect a severely disturbed membrane insertion process in these two mutant toxins, leading to reduced pore function and toxicity. The results support the model that domain I is involved in membrane integration and pore formation.
Phosphorylation of p27Kip1 by Epstein-Barr Virus Protein Kinase Induces Its Degradation through SCFSkp2 Ubiquitin Ligase Actions during Viral Lytic Replication*

Epstein-Barr virus (EBV) productive replication occurs in an S-phase-like cellular environment with high cyclin-dependent kinase (CDK) activity. The EBV protein kinase (PK), encoded by the viral BGLF4 gene, is a Ser/Thr protein kinase, which phosphorylates both viral and cellular proteins, modifying the cellular environment for efficient viral productive replication. We here provide evidence that the EBV PK phosphorylates the CDK inhibitor p27Kip1, resulting in ubiquitination and degradation in a proteasome-dependent manner during EBV productive replication. Experiments with BGLF4 knockdown by small interfering RNA and BGLF4 knock-out viruses clarified that EBV PK is involved in p27Kip1 degradation upon lytic replication. Transfection of the BGLF4 expression vector revealed that EBV PK alone could phosphorylate the Thr-187 residue of p27Kip1 and that the ubiquitination and degradation of p27Kip1 occurred in an SCFSkp2 ubiquitin ligase-dependent manner. In vitro, EBV PK proved capable of phosphorylating p27Kip1 at Thr-187. Unlike cyclin E-CDK2 activity, the EBV PK activity was not inhibited by p27Kip1. Overall, EBV PK enhances p27Kip1 degradation effectively upon EBV productive replication, contributing to establishment of an S-phase-like cellular environment with high CDK activity.
Novel Dextranase Catalyzing Cycloisomaltooligosaccharide Formation and Identification of Catalytic Amino Acids and Their Functions Using Chemical Rescue Approach*

Background: Catalytic residues and molecular mechanism of GH-66 enzymes were hitherto unknown. Results: Novel dextranase produced isomaltotetraose and cyclo-isomaltosaccharides. Its nucleophile (Asp340) and acid/base catalyst (Glu412) were identified by a chemical rescue approach. Conclusion: Three GH-66 enzyme types were newly classified for the first time. Significance: This work elucidates production of isomaltotetraose and cycloisomaltosaccharides, classification of GH-66, identification of catalytic residues, and novel dextran-forming type chemical rescue. A novel endodextranase from Paenibacillus sp. (Paenibacillus sp. dextranase; PsDex) was found to mainly produce isomaltotetraose and small amounts of cycloisomaltooligosaccharides (CIs) with a degree of polymerization of 7–14 from dextran. The 1,696-amino acid sequence belonging to the glycosyl hydrolase family 66 (GH-66) has a long insertion (632 residues; Thr451–Val1082), a portion of which shares identity (35% at Ala39–Ser1304 of PsDex) with Pro32–Ala755 of CI glucanotransferase (CITase), a GH-66 enzyme that catalyzes the formation of CIs from dextran. This homologous sequence (Val837–Met932 for PsDex and Tyr404–Tyr492 for CITase), similar to carbohydrate-binding module 35, was not found in other endodextranases (Dexs) devoid of CITase activity. These results support the classification of GH-66 enzymes into three types: (i) Dex showing only dextranolytic activity, (ii) Dex catalyzing hydrolysis with low cyclization activity, and (iii) CITase showing CI-forming activity with low dextranolytic activity. The fact that a C-terminal truncated enzyme (having Ala39–Ser1304) has 50% wild-type PsDex activity indicates that the C-terminal 392 residues are not involved in hydrolysis. GH-66 enzymes possess four conserved acidic residues (Asp189, Asp340, Glu412, and Asp1254 of PsDex) of catalytic candidates. Their amide mutants decreased activity (11,500 to 140,000 times), and D1254N had 36% activity. A chemical rescue approach was applied to D189A, D340G, and E412Q using α-isomaltotetraosyl fluoride with NaN3. D340G or E412Q formed a β- or α-isomaltotetraosyl azide, respectively, strongly indicating Asp340 and Glu412 as a nucleophile and acid/base catalyst, respectively. Interestingly, D189A synthesized small sized dextran from α-isomaltotetraosyl fluoride in the presence of NaN3.
Predicting Youth Life Leadership Skills Development among Senior 4-H Members: A Tri-State Study.

Leadership development has been and continues to be a major goal of most youth programs. The 4-H youth development program of the Cooperative Extension Service is no exception. The 4-H program states its mission as, “To help youth and volunteers in their development through educational programs using the knowledge base of the land grant universities of the United States (United States Department of Agriculture, 1986, p. 4). " Commitment to the mission involves support of specific program objectives including helping youth develop leadership capabilities, personal standards and values, positive selfconcepts and effective communication skills. (United States Department of Agriculture, 1986, p. 5). As the world enters the 21st century, many youth programs, including 4-H are focusing on the effectiveness of their leadership training. A general perception prevails that participation in a variety of activities or programs such as public speaking or holding office develops leadership life skills and self-understanding. The question remains How accurate is this perception?
Biocatalysts in Control of Phytopatogenic Fungi and Methods for Antifungal Effect Detection

The diseases in plants can be reduced with chemical control, in the case of plant fungi illness can be controlled using fungicides. Chemical fungicides are toxic substances that are used to prevent growth or kill fungi harmful to plants, animals or humans. Their use presents many environmental problems such as: the development of resistance to insecticides in pest populations, the resurgence of the populations treated, chemical waste accumulation, risks and legal complications, destruction of beneficial species. Moreover, it is need to be taking in consideration high cost of fumigants, equipment, labor work and material. Thus, to minimize an environmental damage it is very important to substitute chemical control by the biological control. Biocontrol means to use natural enemies or their metabolites against pathogens causing diseases. This method provides a decrease of pathogenic microorganisms’ population. The main advantages are: little or no adverse side effects to the other biological systems different from pathogenic microorganisms; rare resistance of pathogenic forms to biological control; not harmful to environment; the favorable relation cost vs benefit; prevention of food contamination by chemicals. Biocontrol methods apply to regulate phytopathogenic fungi growth by using an antagonistic microorganism (fungi or bacteria) and / or its derivatives as an active ingredient. These biofungicides, called like that due to its biological origin, are easily assimilated into the environment and they are alternative techniques to replace the use of chemicals in agriculture. Development of new biocontrol approaches and biofungicides, as well as methodologies for detection of antifungal activity are the one of the important goals of biotechnology. A fungal disease can be described as polycyclic whether the causative agent is capable of producing spores and re-infect plants during a growing season or monocyclic when the causative agent must wait for a new season. This classification applies to regions with four seasons where pathogens must produce specific structures to survive the winter. Phytopathogenic fungi can also be distinguished by the types of produced spores and the method by which they penetrate into the plant. Once the pathogen has penetrated, it
The Credit Default Swap Market's Reaction to Earnings Announcements

This paper examines the efficiency of the CDS market by conducting a comparative event study in which both the CDS and the stock markets' responses to earnings announcements are considered. I find that both markets have statistically significant reactions to earnings announcements and both markets anticipate these informational events up to 90 trading days prior to announcement. I further find that neither markets' reaction to earnings announcements is entirely efficient as there is evidence of both over- and under-reaction to earnings news. However, results are sensitive to both the categorization of earnings and the model used to generate abnormal performance.
5-Hydroxytryptamine 1A and 2B Serotonin Receptors in Neurite Outgrowth: Involvement of Early Growth Response Protein 1

Neurotransmitters play important roles in neurogenesis; in particular, acetylcholine and serotonin may regulate neurite elongation. Acetylcholine may also activate transcription factors such as early growth response protein 1 (EGR-1), which plays a role in neurite extension. N18TG2 neuroblastoma cells (which do not produce neurotransmitters and constitutively express muscarinic acetylcholine receptors) were transfected with constructs containing the cDNA for choline acetyltransferase, 5-hydroxytryptamine 1A (5-HT1A) and 5-HT2B serotonin receptors to study acetylcholine and serotonin interplay in neurite outgrowth. 5-HT1A receptor stimulation causes a decrease in EGR-1 levels and inhibition of neurite outgrowth; 5-HT2B stimulation, however, has no effect. Muscarinic cholinergic stimulation, on the other end, increases EGR-1 levels and fiber outgrowth. Inhibition of EGR-1 binding reduces fiber outgrowth activity. When both cholinergic and 5-HT1A receptors are stimulated, fiber outgrowth is restored; therefore, acetylcholine counterbalances the inhibitory effect of serotonin on neurite outgrowth. These results suggest that EGR-1 plays a role in the interplay of acetylcholine and serotonin in the regulation of neurite extension during development.
Biological effect of far‐infrared therapy on increasing skin microcirculation in rats

Background/purpose: Insufficient microcirculation of skin leads to acute and chronic tissue ischemia in cases of trauma, reconstructive surgery, diabetes mellitus and peripheral arterial occlusive disease. The autonomic nervous system and nitric oxide (NO) play important roles in maintaining blood perfusion of the skin. Far‐infrared (FIR) therapy provides low energy of light emitted from an artificial radiator and has been used to treat many vascular‐related disorders. Nevertheless, the mechanisms through which FIR works remain unclear. The present study aims to test the hypothesis that the effect of FIR is through increasing skin microcirculation by a mechanism other than its thermal effect.
On the verification problem of nonregular properties for nonregular processes

Investigate the verification problem of infinite-state processes w.r. t. nonregular properties, i.e. nondefinable by finite-state /spl omega/-automata. We consider processes in the algebra PA (Process Algebra) which provides sequential and parallel (merge) composition, nondeterministic choice and recursion. The algebra PA integrates and strictly subsumes the algebras BPA (Basic Process Algebra, i.e. context-free processes) and BPP (Basic Parallel Processes). On the other hand, we consider properties definable in a new temporal logic called CLTL (Constrained Linear-Time Logic) which is an extension of the linear-time temporal logic LTL with two kinds of constraints on traces: constraints on the numbers of occurrences of states expressed using Presburger formulas (occurrence constraints), and constraints on the order of appearance of states expressed using finite-state automata (pattern constraints). Pattern constraints allow to capture all the /spl omega/-regular properties whereas occurrence constraints allow to define nonregular properties. Then, we present (un)decidability results concerning the verification problem for the different classes of processes mentioned above and different fragments of CLTL.
Does evidence of network effects on firm performance in pooled cross‐section support prescriptions for network strategy?

Strategic prescriptions drawn from pooled cross-sectional evidence of firm performance effects are not necessarily warranted. This is because firm characteristics can influence both the mean and variance of firm performance. Strategic inferences are warranted if empirically observed effects reflect increases in mean firm performance. If they reflect increases in firm performance variance, however, such inferences are warranted only if the increased odds of achieving high performance compensate sufficiently for the concomitantly increased risk of realizing poor performance. Our simulation study, which contrasts firm performance effects in pooled cross-section and within-firm over time, counsels caution when basing strategic prescriptions on pooled cross-sectional studies of firm performance in general, and in the case of network effects in particular.
Corporate governance and performance of small and medium sized enterprise (SME): evidence from Bangladesh

This study examines corporate governance mechanisms and firm performance of medium and small sized enterprises (SMEs) in Bangladesh. From an observation of 769 firm years it is found that there is a significant positive relationship between the following: largest block holding and firm performance; executives pay and firm performance; and debt ratio and firm performance under market based performance measure in Bangladeshi SMEs. While it is found that there is a significant negative relationship between debt to equity ratio and firm performance under ROA performance measure. However, it is vindicated that there is no relationship between other corporate governance variables with firm performance of SMEs under any of the measures. The findings support mostly the principal-agency relationship instead that of the theory on stewardship principles. The findings imply that the underlying differences among SMEs give rise to a further consideration for good corporate governance practices. As well, the ramification of this study can be considered an extension to the literature on SMEs in the context of a less developed country.
Examination of intra and interrater reliability with a new ultrasonographic reference atlas for scoring of synovitis in patients with rheumatoid arthritis

Objective Synovitis in patients with rheumatoid arthritis (RA) may be scored semiquantitatively (0–3) for B-mode (BM) and power Doppler (PD) ultrasonography. The objective was to assess the reliability of BM and PD examinations with a novel ultrasonographic atlas as reference. Methods Representative ultrasound images (including scores 0–3) of BM and PD from 24 different joints were collected to develop an ultrasonographic atlas. Ten RA patients were assessed twice by five rheumatologists performing BM and PD scoring (0–3) of 16 joints bilaterally (metacarpophalangeal 1–5, wrist (radiocarpal, intercarpal, radioulnar), elbow, knee, talocrural and metatarsophalangeal 1–5), with the novel ultrasonographic atlas as a reference. Results The median (range) percentages of exact agreements for BM/PD assessments were 73.1 (70.3–80.6)/83.7 (76.7–87.6) and for close agreement 98.1 (96.2–99.7)/98.0 (96.8–98.4) with weighted κ values of median (range) 0.77 (0.70–0.83) for BM and 0.83 (0.73–0.86) for PD. The intrarater intraclass correlation coefficients (ICC) for BM/PD scores were 0.95 (0.93–0.99)/0.97 (0.95–0.99) and interrater ICC were 0.95 (0.86–0.99)/0.97 (0.94–1.00). Scoring of 32 joints was completed in median 15 min (range 12–20). Conclusion With the use of an ultrasonographic atlas as reference high intra and interrater reliability was found for BM and PD scoring. This novel atlas may be a useful resource in clinical practice and research.
Safety and immunogenicity of bacterial and tobacco plant cell line derived recombinant native and mutant Escherichia coli heat-labile toxin in chickens

The safety and immunogenicity of the mammalian mucosal adjuvants, Escherichia coli wild-type heat-labile holotoxin (LT) and E. coli mutant LT (LTA-K63/LTB), were examined in 1-day-old chicks and 10-day-old to 21-day-old broilers. Biologically active, E. coli recombinant wild-type LT and recombinant LTA-K63/LTB produced in a transgenic Nicotiana tabacum (NT-1) tobacco cell line (SLT102) were tested for safety and antigenicity following various routes of administration. Safety was assessed by clinical signs, body weight gain, gross organ pathology and wet organ weight, and histopathology. Antigenicity was assessed using LT-B-specific serum IgG enzyme-linked immunosorbent assay. Parenteral administration of E. coli recombinant wild-type LT did not have any discernible effect on bird health and was well tolerated at levels up to 400 µg per dose. Recombinant, SLT102-derived mutant LT derived from SLT102 cells retained in vitro ganglioside binding and was safe and antigenic following repeated mucosal administration to birds. The highest systemic LT-B-specific IgG titres were detected in birds that received three on-feed doses of SLT102-derived mutant LT. Among the various SLT102-derived mutant LT preparations tested, whole, wet cells or whole cell lysates were the most antigenic. These results demonstrate for the first time that E. coli-derived recombinant, wild-type LT holotoxin is well tolerated following multiple administrations to young birds at body weight doses previously reported to be enteropathogenic and toxic in mammalian species. Moreover, these data also demonstrate the feasibility of using recombinant wild-type and mutant LT produced in transgenic NT-1 tobacco cells as safe and potent vaccine adjuvants in poultry.
Interlaboratory Study of Free Cyanide Methods Compared to Total Cyanide Measurements and the Effect of Preservation with Sodium Hydroxide for Secondary‐ and Tertiary‐Treated Waste Water Samples

Several methods exist for the measurement of cyanide levels in treated wastewater,typically requiring preservation of the sample with sodium hydroxide to minimize loss of hydrogen cyanide gas (HCN). Recent reports have shown that cyanide levels may increase with chlorination or preservation. In this study, three flow injection analysis methods involving colorimetric and amperometric detection were compared within one laboratory, as well as across separate laboratories and equipment. Split wastewater samples from eight facilities and three different sampling periods were tested. An interlaboratory confidence interval of 3.5 ppb was calculated compared with the intralaboratory reporting limit of 2 ppb. The results show that free cyanide measurements are not statistically different than total cyanide levels. An artificial increase in cyanide level is observed with all methods for preserved samples relative to nonpreserved samples, with an average increase of 2.3 ppb. The possible loss of cyanide without preservation is shown to be statistically insignificant if properly stored up to 48 hours. The cyanide increase with preservation is further substantiated with the method of standard additions and is not a matrix interference. The increase appears to be correlated with the amount of cyanide observed without preservation, which appears to be greater in those facilities that disinfect their wastewater with chlorine, followed by dechlorination with sodium bisulfite.
Combination antiretroviral therapy and duration of pregnancy

ObjectiveTo assess the association between type and timing of initiation of antiretroviral therapy in pregnancy and duration of pregnancy. DesignProspective study. MethodsData on 3920 mother–child pairs were examined (3015 mother–child pairs from the European Collaborative Study and 905 from the Swiss Mother + Child HIV Cohort Study). Factors examined included gestational age, antiretroviral therapy during pregnancy, maternal CD4 count, viral load, illicit drug use (IDU) and mode of delivery. Deliveries at less than 37 weeks were defined as premature. ResultsThe prematurity rate was 17% and median gestational age 39 weeks. Twenty-three per cent (896 of 3920) of women received antiretroviral therapy during pregnancy: 64% (573 of 896) zidovudine monotherapy, 24% (215) combination therapy without protease inhibitors (PI) and 12% (108) combination therapy with PI. In multivariate analysis, adjusted for maternal CD4 count and IDU, odds ratio (OR) of prematurity was 2.60 [95% confidence interval (CI), 1.43–4.75] and 1.82 (95% CI, 1.13–2.92) for infants exposed to combination therapy with and without a PI, respectively, compared to no treatment. Exposure to monotherapy was not associated with prematurity, but severe immunosuppression and IDU in pregnancy were. Women on combination therapy from before pregnancy were twice as likely to deliver prematurely as those starting therapy in the third trimester (OR, 2.17; 95% CI, 1.03–4.58). ConclusionsPregnancy issues should be discussed when making decisions about initiation of combination antiretroviral therapy for HIV-infected women. Elective caesarean section to reduce vertical transmission at 36 weeks rather than 38 weeks may be advisable in women on combination therapy with PI.