Journal articles on the topic 'IF3'

Abstract Initiation factor IF3 is an essential protein that enhances the fidelity and speed of bacterial mRNA translation initiation. Here, we describe the dynamic interplay between IF3 domains and their alternative binding sites using pre-steady state kinetics combined with molecular modelling of available structures of initiation complexes. Our results show that IF3 accommodates its domains at velocities ranging over two orders of magnitude, responding to the binding of each 30S ligand. IF1 and IF2 promote IF3 compaction and the movement of the C-terminal domain (IF3C) towards the P site. Concomitantly, the N-terminal domain (IF3N) creates a pocket ready to accept the initiator tRNA. Selection of the initiator tRNA is accompanied by a transient accommodation of IF3N towards the 30S platform. Decoding of the mRNA start codon displaces IF3C away from the P site and rate limits translation initiation. 70S initiation complex formation brings IF3 domains in close proximity to each other prior to dissociation and recycling of the factor for a new round of translation initiation. Altogether, our results describe the kinetic spectrum of IF3 movements and highlight functional transitions of the factor that ensure accurate mRNA translation initiation.

Initiation of translation involves the assembly of a ribosome complex with initiator tRNA bound to the peptidyl site and paired to the start codon of the mRNA. In bacteria, this process is kinetically controlled by three initiation factors—IF1, IF2, and IF3. Here, we show that deletion of helix H69 (∆H69) of 23S rRNA allows rapid 50S docking without concomitant IF3 release and virtually eliminates the dependence of subunit joining on start codon identity. Despite this, overall accuracy of start codon selection, based on rates of formation of elongation-competent 70S ribosomes, is largely uncompromised in the absence of H69. Thus, the fidelity function of IF3 stems primarily from its interplay with initiator tRNA rather than its anti-subunit association activity. While retaining fidelity, ∆H69 ribosomes exhibit much slower rates of overall initiation, due to the delay in IF3 release and impedance of an IF3-independent step, presumably initiator tRNA positioning. These findings clarify the roles of H69 and IF3 in the mechanism of translation initiation and explain the dominant lethal phenotype of the ∆H69 mutation.

Novel therapeutic approaches are much needed for the treatment of osteosarcoma. Targeted radionuclide therapy (TRT) and radioimmunotherapy (RIT) are promising approaches that deliver therapeutic radiation precisely to the tumor site. We have previously developed a fully human antibody, named IF3, that binds to insulin-like growth factor 2 receptor (IGF2R). IF3 was used in TRT to effectively inhibit tumor growth in osteosarcoma preclinical models. However, IF3’s relatively short half-life in mice raised the need for improvement. We generated an Fc-engineered version of IF3, termed IF3δ, with amino acid substitutions known to enhance antibody half-life in human serum. In this study, we confirmed the specific binding of IF3δ to IGF2R with nanomolar affinity, similar to wild-type IF3. Additionally, IF3δ demonstrated binding to human and mouse neonatal Fc receptors (FcRn), indicating the potential for FcRn-mediated endocytosis and recycling. Biodistribution studies in mice showed a higher accumulation of IF3δ in the spleen and bone than wild-type IF3, likely attributed to abnormal spleen expression of IGF2R in mice. Therefore, the pharmacokinetics data from mouse xenograft models may not precisely reflect their behavior in canine and human patients. However, the findings suggest both IF3 and IF3δ as promising options for the RIT of osteosarcoma.

A Bacillus stearothermophilus in vitro translational system has been developed to study the expression of the three cistrons (infC, rpmI, and rplT) constituting the infC operon of this bacterium. When directed by homologous in vitro transcribed infC tricistronic mRNA, this system, which consists of partially purified and purified components of the B. stearothermophilus translational apparatus, synthesizes with high efficiency and specificity the three gene products (IF3, L35, and L20) in a ratio similar to that found in vivo (i.e., about 1:6:6). The three cistrons are translationally coupled and expressed in a specific temporal order: a low level of IF3 synthesis stimulates the expression of L35 which, in turn, greatly stimulates the synthesis of L20 and IF3. Protein L20 and an excess of IF3 were found to act as translational feedback inhibitors of the entire operon. The synthesis of IF3 displayed a strong dependence on IF2. This dependence as well as the repressibility by excess IF3 were found to be due to the presence of the rare AUU initiation triplet at the beginning of infC.Key words: translational coupling, IF3, IF2, L35, L20.

ABSTRACT A primer extension inhibition (toeprint) assay was developed using ribosomes and ribosomal subunits from Streptomyces lividans. This assay allowed the study of ribosome binding to streptomycete leaderless and leadered mRNA. Purified 30S subunits were unable to form a ternary complex on aph leaderless mRNA, whereas 70S ribosomes could form ternary complexes on this mRNA. 30S subunits formed ternary complexes on leadered aph and malE mRNA. The translation initiation factors (IF1, IF2, and IF3) from S. lividans were isolated and included in toeprint and filter binding assays with leadered and leaderless mRNA. Generally, the IFs reduced the toeprint signal on leadered mRNA; however, incubation of IF1 and IF2 with 30S subunits that had been washed under high-salt conditions promoted the formation of a ternary complex on aph leaderless mRNA. Our data suggest that, as reported for Escherichia coli, initiation complexes with leaderless mRNAs might use a novel pathway involving 70S ribosomes or 30S subunits bound by IF1 and IF2 but not IF3. Some mRNA-ribosome-initiator tRNA reactions that yielded weak or no toeprint signals still formed complexes in filter binding assays, suggesting the occurrence of interactions that are not stable in the toeprint assay.

<strong>Introduction</strong> Okra (<em>Abelmoschus esculentus</em> L. Moench) is a perennial vegetable of African origin, with production areas expanding throughout the tropical, sub-tropical and warm temperate regions of the world (Benchasri, 2012). Okra is a versatile crop produced forits pods, leaves, seedoil and protein, gums, and fiber in different parts of the world (Lamont,1999). It is produced in Vilanculos primarily for the immature pods, though leaves are consumed to a limited extent in rural communities. Important attributes of okra pods are shape and color, earliness, and totalmarketable yield. Since okra is handharvested, plant height and architecture,as well as the absence of spines are important tofacilitate harvesting (Simonne, et al, 2012).Okra is not only reach in nutrients (fats, proteins, carbohydrates, minerals and vitamins), but it is also believed to have medicinal properties. The climate in much of Mozambique presents favorable conditions for the production of Okra (INE, 2011) and because of the growing preference for Okra among consumers in Mozambique, its area of production had rapidly expanded to every part of the country, mainly in the provinces of Tete, Manica, Sofala and Inhambane. Okra provides a good alternative or supplemental income for smallholder farmers (COSTA <em>et al</em>., 1981) and its production in the district of Vilanculos requires the application of considerable amount of irrigation water to improve productivity. Small farmers-led irrigation in Mozambique takes place in diverse forms, including bucket irrigation, sprinkler and drip systems, furrow and small pumped irrigation systems. However, Mozambique has limited access to raw water supplies and the country as a whole is extremely vulnerable (ranks 3^rd amongst African countries) to extreme weather patterns (i.e. recurring droughts and flood events), which contribute to crop instability, food insecurity and malnutrition (USAID, 2010). However, information on watering (irrigation) frequencies for optimal growth and development of this valuable crop under the edaphic-climatic conditions in the district of Vilanculos is lacking. Climate-smart strategies to increase irrigation efficiency at small-farm level are essential in helping rural smallholders optimize production of okra and other vegetable crops under climate change. <strong>Materials and Methods</strong> <strong>1. Geographical location</strong> The field experiment was carried out between October and December 2016 on the campus of the Universidade Eduardo Mondlane-Escola Superior de Desenvolvimento Rural (UEM-ESUDER) located in the coastal town of Vilanculo within the district of Vilanculos, Province of Inhambane in Mozambique at coordinates 21&ordm;59&rsquo;30.6&rdquo; S, 035&ordm;16&rsquo;14.8&rdquo; E. and an average altitude of 49m. The district of Vilanculosoccupies the northern part of the province of Inhambane, bordering the district of Inhassoro to the north, the district of Massinga to the south, the districts of Mabote and Funhalouro to the west and the Indian Ocean to the east (MAE, 2005). <strong>2. Climatic condition</strong> The climate of the study area is Aw according to the K&ouml;eppen e Geiger, which corresponds to a dry tropical climate with two distinct seasons. A wet season spans from October to March, with an average annual precipitation of 1300 mm; and a dry season from April to September, with an average annual precipitation of 700 to 900 mm (MICOA, 2009 and MAE, 2005).Figure-1 illustrates the average temperature and precipitation patterns in Vilanculos. The driest month is July, which averages 17 mm of rainfall, and the month of February is the wettest with an average precipitation of 166 mm. On average, the temperatures are always high, with an annual average of 24^oC. <strong>3</strong>. <strong>Soils of the study area</strong> The soils in the district of Vilanculos are sandy and permeable in the coastal areas, and sandy-loam to loamy-clay in the interior. The study site was approximatively 10 km from the coastal line and its soils are predominantly sandy and permeable with low organic content (8.55%) and low water holding capacity (less than 5cm/m) (MAE, 2005; Maite, 2014). The growing season averages 120 to 149 days, and because of low precipitation and recurring drought periods during the growing seasons, the area&rsquo;s potential for rain-fed agriculture is marginal (Mafalacusser, 2013.) <em>Figure 1. Monthly temperature and precipitation patterns in the district of Vilanculo, Mozambique.</em> <em>Adapted from Climograma Vilanculos: </em><em>https://pt.climate-data.org/location/52395/</em> &nbsp; <strong>4. Experimental design</strong> The experiment was laid out on an area of 83.375m² within which 3 blocks of equal sizes were setup, each block divided into 4 parcels of 4 m² (2 m x 2 m) for a total of 12 parcels. &nbsp;The blocks were setup at 1m apart and the parcels within each block were spaced at 50cm. Treatments were then randomly assigned to each parcel, resulting in each parcel representing a unique treatment within each block. Table-1 describes the different treatments (irrigation frequencies) used in the experiment. <em>Table 1. Treatments details: Four (4) treatments randomly assigned to the experimental blocks.</em> IF1 <em>Irrigate twice a day in the mornings (5-6 AM), and afternoons (4-5 PM)</em> IF2 <em>Irrigate once a day in the afternoons (4-6PM)</em> IF3 <em>Irrigate once a day in the mornings (5-6 AM)</em> IF4 <em>Irrigate twice a day in the mornings (5-6 AM), and afternoons (4-5 PM) at 1- day intervals.</em> <strong>5. Soil preparation and planting</strong> Soil preparation consisted of removing natural vegetation from the experimental site with a manual hoe without revolving the top soil, and the soil surface was raked and leveled prior to subdivision of the experimental area into parcels, blocks and replications for the study.Seeds of okra (<em>Abelmoschus esculentus</em> L. Moench) variety Clemson Spineless, were soaked in water for 24 hours to stimulate germination, and then sown by placing two (2) seeds in each planting hole at an approximate depth of 5cm in each parcel. Fifteen (15) days following the emergence, the seedlings were thinned by removing the least developed plant in each planting hole, leaving only one (1) okra seedling per hole in each treatment. The seedling selection criteria was based on height, and the number of developed leaves. <strong>6. Soil fertilization</strong> Organic fertilizer in the form of bovine manure was applied seven (7) days prior to sowing and 15 days after seedling emergence, at the rate of 15 tons/ha each. All plants received the same fertilizer application. <strong>7. Irrigation</strong> A hand-held irrigation bucket was used for applying water to the experimental plots, a watering scheme consistent with the most popular irrigation system among smallholders in the study area. All parcels were uniformly irrigated with 13 liters of water, starting from one (1) day prior to seeding until fifteen (15) days after seeding when the okra plants developed up to three (3) fully expanded leaves. Starting from day-15 through the end of the experiment, plants in each parcel were irrigated according to the established watering schedule (Table-1.) <strong>8.</strong> <strong>Pests/Weed Control</strong> Weed control was manual and continuous throughout the study, by hand plucking at the onset of their appearance to avoid perturbation of soil and plants. The most common weeds were <em>amaranthus</em>, <em>commelina benghalensis</em> and cyperus<em> esculentus</em> species, with <em>amaranthus</em>and<em>cyperus esculentus</em> being the most dominants.Observed pests were acarids and caterpillars, and these were treated with pesticides Cipermetrina 25% E.C. and Fords. The occurrence of fungi during the experiment was sparse, but fungi was treated with the fungicide Bravo. <strong>Data Collection</strong> <strong>1. Pre-harvest Data&nbsp;&nbsp; </strong> Pre-harvest plant parameters consisted of plant height and stem diameter. Plant height was determined by measuring the height of the primary stem of random sample of plants in each treatment from the base at soil surface to the apex, using a metric ruler. At the same time, stem diameter was measured with a digital Caliper, within 300mm at 5 cm above the soil surface. Growth Rate of okra plants was determined from height measurements and expressed as <em>Absolute Growth Rate</em> (AGR), based upon increments of measured plant heights between sampling dates (<em>i.e. Height at day2- Height at day1</em>) over the time-period (<em>number of days</em>) between measurement dates. <strong>2. Post-harvest Data</strong> Post-harvest data included Fruit Length, Fruit Diameter, and Fruit Weight. Okra fruits were harvested twice; the first harvest took place 90 days after seeding, and the second harvest 8 days later, or 98 days after seeding. Harvest data were from random fruit samples under each treatment for both harvests. A graduated metric rulerwas used to measure fruit length, while fruit diameter was measured with a digital Caliper within at 5 cm from thebase of each fruit, and fruit weight was determined by weighing sample fruits per treatment from each harvest, with a digital scale equipped with a high precision strain gauge sensor system. <strong>Data Analyses&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp; </strong> All data collected from the experiment were analyzed with the statistical software SPSS(v.20, IBM SPSS Chicago). Analyses of Variance (ANOVA) were performed to determine differences between treatments, followed by mean comparisons based on Tukey HSD (&alpha; = 0.05). Homogeneity of the variances was verified through the test of Levene (&alpha; = 0.05), and graphical illustrations were produced with the Microsoft Excel program. <strong>Result and Discussion</strong> <strong>1. Plant Height</strong> The different irrigation treatments had no significant effect on plant height at each sampling dates. Okra plants grown in this experiment averaged 4.73cm at 7 days after sowing (DAS), and increased through the study to an average of 49.39 cm at 63 DAS (Fig.2.)&nbsp; In contrast with other studies, okra plant height at the end of this experiment was relatively shorter than heights observed elsewhere. For example, Saifullah and Rabbani (2009) reported from evaluation of different genotypes that okra plant height at final harvest ranged from 81.80cm to 196.17cm. It is probable that the conditions under which the present study was carried out did not favor greater shoot elongation of the okra plants. However, significant differences in plant heights were observed between sampling dates according to the growth progression of plants through the study period, and between treatments across sampling dates. As illustrated in Figure 2, plant heights during the first three (3) sampling dates (7-21 DAS) were significantly lower than plant heights at other sampling dates. <em>Figure 2. Average plant height per treatment per sampling date</em> &nbsp; Furthermore, plant heights at 28-35 DAS was significantly lower than heights at 63 DAS. Overall, plant heights continuously increased during the study period, except under IF2 (<em>watering once a day in the afternoons</em>), where plant height leveled off between 28 and 35 DAS, before increasing again through the end of the study period (Fig.2.) Starting from 35 DAS to 63 DAS, okra plants height under IF2 increased rapidly to match heights of plants under IF1 and surpass heights of plants under IF3 and IF4 at 63 DAS. Figure-3 further shows okra plant height averaged across treatments at each sampling date.&nbsp; Okra plant height followed a rather exponential pattern from 7 DAS until 35 DAS, and then the growth pattern became linear thereafter. Significant differences in plant heights were observed during the first phases of plant growth between 7 DAS and 21 DAS, and between the set of 7 DAS - 21 DAS and the remainder DAS. However, no significant differences in plant height were observed between sampling dates from 28 DAS through 63 DAS (Fig.3.) &nbsp; &nbsp; <em>Figure 3.Average plant height per sampling date across the treatments</em> &nbsp; Figure 4 illustrates the average plant height per treatment across sampling dates. Plants grown under IF3 (<em>watering once a day in the mornings</em>) were significantly taller (P&lt;0.05) than those under IF2 and IF4. Although watering plants twice a day (IF1) resulted in plant heights similar to those under IF3, these results suggest that in terms of okra plant growth, watering once a day in the mornings (IF3) was more efficient in promoting plant growth with less use of irrigation water, compared with IF1 (<em>watering twice a day</em>.) <em>Figure 4.Average plant height per treatment across sampling dates</em> &nbsp; Moreover, plants grown under IF4 which received irrigation water twice a day but at one-day intervals, significantly outperformed those under IF2 (<em>watering once a day in the afternoons</em>) in terms of plant height. Given equal amounts of irrigation water applied but at different intervals in or IF2 and IF4, the treatment IF4 appeared to be more efficient in promoting okra plant growth as compared with IF2 (Fig.4.) <strong>2. Plant Growth Rate&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp; </strong> Growth Rate of okra plants was determined from height measurements and expressed as <em>Absolute Growth Rate</em> (AGR), based upon increments of measured plant heights between sampling dates (<em>i.e. Height at day2- Height at day1</em>) over the time-period (<em>number of days</em>) between measurement dates.&nbsp; Figure 5 illustrates the AGR pattern of okra plants under each treatment at each sampling date, with dates expressed in number of days after sowing (DAS). <em>Figure 5. Average Plant Growth Rate (cm/day) per sampling date across the treatments</em> &nbsp; As illustrated in Fig.5, okra plants grown under IF1 and IF3 exhibited relatively similar AGR pattern during much of the study period, while those under IF2 and IF4 constitute the other pair of similar AGR. Averaged across treatments, the AGR of okra plants under this study was 0.568 cm/day at 14 DAS and then increased significantly (P&lt;0.05) to a maximum rate of 2.346 cm/day at 28 DAS. After 28 days or so of growth have passed, the AGR dropped to a low of 0.309 cm/day at 35 DAS, before leveling off to an average rate 0.536 cm/day for the remainder of the growth period (Fig.6.)&nbsp; <em>Figure 6. Average Growth Rate per sampling date across treatments</em> &nbsp; Not many studies appear to have been carried out on okra growth rate, but the AGR pattern exhibited under this study was similar to those observed in previous experiments (Hunt, 1978; Tarassoum and Lovane, 2019.) Similar to previous studies, the reduction in AGR after 28 DAS have passed was attributable to Phenology and/or resource allocation to functions other than plant height.Comparisons of AGR between treatments across sampling dates showed that the average AGR was significantly faster (P&lt;0.05) under IF2 and IF4 when compared with IF1 and IF3 (Fig.7), with no significant differences observed between the respective pairs of treatments.Strikingly, the two treatments (IF1 &amp; IF3) which produced taller plants (Fig.4) resulted in relatively lower overall AGR during the study period, compared with the pair of IF2 and IF4, which resulted in shorter plants. While plant height under IF2 and IF4 were lower during the first 21 days and after 35 DAS have passed (Fig. 3), the significantly rapid increase in their heights between 21 and 28 DAS resulted in an overall greater AGR compared with the other treatments. <em>Figure 7. Average Growth Rate per treatment across sampling date</em> &nbsp; &nbsp; <strong>3. Plant Diameter</strong> Table 5 presents the average plant diameter under each treatment. The different irrigation frequencies had no significant effects on okra plant diameter, which averaged from 7.83 cm to 7.86 cm across the treatments. <em>Table 5. Average plant diameter per treatment across sampling dates</em> Treatments (Irrigation Frequency) Average Plant Diameter (cm) IF1 7.83&ordf; IF2 7.83&ordf; IF3 7.86&ordf; IF4 7.83&ordf; <strong>Average Treatments</strong> <strong>7.85</strong> <em>Means followed by the same letters are not significantly different (P&gt;=0.05)</em> The average stem diameter of okra plants under IF3 was slightly above the diameter under other treatments, but the difference was not significant (P&gt;0.05.) However, like plant height, plant diameter also increased exponentially from an average of 2.5cm on the first sampling date (7 DAS), to about 10.0cm on the fourth sampling date (28 DAS) and remained constant for the remainder of the study period (Fig.8.)&nbsp; <em>Figure 8: Average plant diameter per sampling date across treatments</em> <strong>4. Fruit Characteristics</strong> Summarized in Table-6 are the okra fruit characteristics under the different irrigation frequencies. Fruit diameter, length and weight under this experiment averaged 0.1852 cm, 11.83 cm and 0.04625 kg per pod across the treatments. Mateus (2011) reported that fruit characteristics of Okra (<em>Abelmoschus Esculentus</em>) Clemson variety, averaged 1.7 cm in diameter, 7.5 cm in length and 10g in weight. Other studies reported fruit diameter from 1.26 to 2.86cm, fruit length from 5.46 to 17.25 cm, and individual fruit weights from 0.01528 to 0.02615kg (Owolarafe and Shotonde, 2004; Saifullah and Rabbani, 2009.) Considering data from these previous studies, it appeared that okra plants under the present experiment were shorter but produced heavier and taller fruits compared with the above referenced study results. The variability in okra fruit characteristics is attributable to genotypes, experimental designs or climate variances amongst study areas. <em>Table 6. Average fruit length, diameter and weight under different irrigation frequencies.</em> IRRIGATION FREQUENCIES VARIABLES Fruit Length (cm) Fruit Diameter (cm) Fruit Weight (kg) IF1 13.0208a 0.2000a 0.060a IF2 11.3542b 0.1913ab 0.041ab IF3 11.2417b 0.1704b 0.046ab IF4 11.7083 b 0. 1813b 0.038b <strong>AVERAGE</strong> <strong>11.83125b</strong> <strong>0.1852ab</strong> <strong>0.04625ab</strong> <em>Means followed by the same letters in a column are not significantly different (P&gt;=0.05)</em> As shown on the Table-6, irrigating plants twice a day (IF1) resulted in significantly longer pods (P&lt; 0.05) compared with all the other treatments. IF4 resulted in a slightly longer fruits when compared with plants grown under IF2 and IF3, but the differences were not statistically significant. Fruit diameter was similar under IF1 and IF2, with no significant differences observed between IF2, IF3 and IF4, although IF4 resulted in a slightly larger fruit diameter than IF3. As for fruit weight, plants grown under IF1, IF2 &amp; IF3 produced fruits of similar weights. The only statistical difference observed was between IF1 and IF4, with the later treatment resulting in significantly lower fruit weight. The results from this study suggest that even though IF1 resulted in longer fruits, the non-significant differences between IF1 &amp; IF2 in terms of fruit diameterand between IF1, IF2 &amp; IF3 in terms of fruit weight, irrigating okra plants once a day (IF2 or IF3) could yield marketable fruits while at the same time reduce the amount of irrigation water. <strong>Conclusions</strong> Okra is produced in the district of Vilanculos primarily for its immature pods, which are either sold in local market or consumed by the producing households. Smallholders comprise the vast majority of okra producers in the study area, growing small plots of various sizes under bucket or furrow irrigation systems to improve growth and yield. Differences in Okra plant characteristics (height and growth rate) under this study appeared to show variation between pairs of treatments, with IF1 &amp; IF3 constituting one pair, and IF2 &amp; IF4 constituting the other pair. Interestingly, the taller plants exhibited lower average growth rate when compared with the shorter plants (Fig.3 &amp; 6.) Differences in fruit characteristics were also observed between plants grown under different watering frequencies. While IF1 resulted in longer okra pods compared with the other treatments, fruit diameter was similar under IF1 and IF2, and fruit weight similar under IF1, IF2 &amp;IF3. These results seemed to suggest that watering okra plants once a day in the mornings (IF3) offer better alternatives for irrigation water-saving strategy for optimal plant height and fruit weight. Additional studies will further assess the once-a-day irrigation schedules at different day-intervals to determine optimal water-saving irrigation schedule for okra production in the district of Vilanculos.

Milk from different cattle breeds can present different casein and fat contents, which are reflected in different cheese yields (CY). However, CY is also related to some breed-related molecular characteristics. The aim of the present work was to quantify the effect of these characteristics by comparing a series of Parmigiano Reggiano (PR) cheese-making trials made with milks from Italian Brown (IB) and Italian Friesian (IF) cattle herds. Twelve trials were carried out in a cheese factory in one year (one trial per month), each one consisting of four vats processed in parallel: three vats contained milk from three different IF cattle herds (IF1, IF2 and IF3) and one contained milk from a single IB cattle herd. A 24-h CY prediction formula was developed with data from IF1, IF2 and IF3 trials (calibration) and successively validated by applying it to 12 PR trials made with IF milk in six different cheese factories (external validation). The predicted values of 24-h CY were no different to the actual ones in both calibration and external validation. Finally, the formula was tested on trials made with IB milk. In this case, the predicted values were lower than the actual ones. The quantity of IF milk casein necessary to give the same CY of IB milk was 0.20 g/100 g.

According to the standard model of bacterial translation initiation, the small ribosomal 30S subunit binds to the initiation site of an mRNA with the help of three initiation factors (IF1–IF3). Here, we describe a novel type of initiation termed “70S-scanning initiation,” where the 70S ribosome does not necessarily dissociate after translation of a cistron, but rather scans to the initiation site of the downstream cistron. We detailed the mechanism of 70S-scanning initiation by designing unique monocistronic and polycistronic mRNAs harboring translation reporters, and by reconstituting systems to characterize each distinct mode of initiation. Results show that 70S scanning is triggered by fMet-tRNA and does not require energy; the Shine–Dalgarno sequence is an essential recognition element of the initiation site. IF1 and IF3 requirements for the various initiation modes were assessed by the formation of productive initiation complexes leading to synthesis of active proteins. IF3 is essential and IF1 is highly stimulating for the 70S-scanning mode. The task of IF1 appears to be the prevention of untimely interference by ternary aminoacyl (aa)-tRNA•elongation factor thermo unstable (EF-Tu)•GTP complexes. Evidence indicates that at least 50% of bacterial initiation events use the 70S-scanning mode, underscoring the relative importance of this translation initiation mechanism.

SUMMARY Valuable information on translation initiation is available from biochemical data and recently solved structures. We present a detailed description of current knowledge about the structure, function, and interactions of the individual components involved in bacterial translation initiation. The first section describes the ribosomal features relevant to the initiation process. Subsequent sections describe the structure, function, and interactions of the mRNA, the initiator tRNA, and the initiation factors IF1, IF2, and IF3. Finally, we provide an overview of mechanisms of regulation of the translation initiation event. Translation occurs on ribonucleoprotein complexes called ribosomes. The ribosome is composed of a large subunit and a small subunit that hold the activities of peptidyltransfer and decode the triplet code of the mRNA, respectively. Translation initiation is promoted by IF1, IF2, and IF3, which mediate base pairing of the initiator tRNA anticodon to the mRNA initiation codon located in the ribosomal P-site. The mechanism of translation initiation differs for canonical and leaderless mRNAs, since the latter is dependent on the relative level of the initiation factors. Regulation of translation occurs primarily in the initiation phase. Secondary structures at the mRNA ribosomal binding site (RBS) inhibit translation initiation. The accessibility of the RBS is regulated by temperature and binding of small metabolites, proteins, or antisense RNAs. The future challenge is to obtain atomic-resolution structures of complete initiation complexes in order to understand the mechanism of translation initiation in molecular detail.

Abstract Recently, there has been a shortage of baby formula due to the COVID-19-related supply chain, unsanitary conditions, and contamination in the manufacturing company. There is genuine concern that contaminated baby formulas may have been inadvertently imported worldwide. For this purpose, five commercially available infant formulas were selected and coded as IF1, IF2, IF3, IF4, and IF5. Physicochemical, nutritional compositions, toxic element (Pb), microbial content, and identification of isolates in the infant formula were determined using standard methods. The pH and total titratable acidity ranged from 6.50±0.00 to 6.90±0.0 and 0.05±0.01 to 0.07±0.00 %, respectively. Moisture, ash, and protein ranged from 2.4±0.1 to 8.00±0.3 %; 1.9±0.01 to 2.7±0.4 %; 1.8±0.0 to 3.8±0.3 %, respectively. Crude fat ranged from 22.0±0.0 to 28.30±0.0 %, and fibre was not detected. Toxic element Pb was detected in all the samples, ranging from 1.2±0.1 to 13.51±0.9 mg/kg. Microbial counts ranged from 2.0×10 to 8.0×104 CFU/g for total viable, enterobacteriaceae 3.0×104 to 13.5×105 CFU/g; staphylococcal 5×103 to 1.0×104 CFU/g and fungal 1.1×10 to 2.4×105 CFU/g. Although the standard is that no pathogen should be in infant formula, several were identified including Klebsiella spp., Escherichia spp., and Staphylococcus spp. In conclusion, the nutritional composition obtained correlated with the product description. Manufacturers should work assiduously on taking steps to ensure the safety of their products by limiting the level of toxic elements and microbial contaminants which negatively affect the health of infants.

Protein biosynthesis in mitochondria is organized in a bacterial manner. However, during evolution, mitochondrial translation mechanisms underwent many organelle-specific changes. In particular, almost all mitochondrial translation factors, being orthologous to bacterial proteins, are characterized by some unique elements of primary or secondary structure. In the case of the organellar initiation factor 3 (IF3), these elements are several dozen amino acids long N- and C-terminal extensions. This study focused on the terminal extensions of baker’s yeast mitochondrial IF3, Aim23p. By in vivo deletion and complementation analysis, we show that at least one extension is necessary for Aim23p function. At the same time, human mitochondrial IF3 is fully functional in yeast mitochondria even without both terminal extensions. While Escherichia coli IF3 itself is poorly active in yeast mitochondria, adding Aim23p terminal extensions makes the resulting chimeric protein as functional as the cognate factor. Our results show that the terminal extensions of IF3 have evolved as the “adaptors” that accommodate the translation factor of bacterial origin to the evolutionary changed protein biosynthesis system in mitochondria.

Background: Osteosarcoma (OS) has an overall patient survival rate of ~70% with no significant improvements in the last two decades, and novel effective treatments are needed. OS in companion dogs is phenotypically close to human OS, which makes a comparative oncology approach to developing new treatments for OS very attractive. We have recently created a novel human antibody, IF3 to IGF2R, which binds to this receptor on both human and canine OS tumors. Here, we evaluated the efficacy and safety of radioimmunotherapy with 177Lu-labeled IF3 of mice bearing canine-patient-derived tumors and performed canine and human dosimetry calculations. Methods: Biodistribution and microSPECT/CT imaging with 111In-IF3 was performed in mice bearing canine OS Gracie tumors, and canine and human dosimetry calculations were performed based on these results. RIT of Gracie-tumor-bearing mice was completed with 177Lu-IF3. Results: Biodistribution and imaging showed a high uptake of 111In-IF3 in the tumor and spleen. Dosimetry identified the tumor, spleen and pancreas as the organs with the highest uptake. RIT was very effective in abrogating tumor growth in mice with some spleen-associated toxicity. Conclusions: These results demonstrate that RIT with 177Lu-IF3 targeting IGF2R on experimental canine OS tumors effectively decreases tumor growth. However, because of the limitations of murine models, careful evaluation of the possible toxicity of this treatment should be performed via nuclear imaging and image-based dosimetry in healthy dogs before clinical trials in companion dogs with OS can be attempted.

ABSTRACT The effect of paromomycin on the interaction of ribosomal subunits was studied. Paromomycin inhibited the antiassociation activity of initiation factor 3 (IF3). Furthermore, ribosomal subunits were associated to form 70S ribosomes by paromomycin even in the presence of 1 mM Mg2+. Paromomycin did not inhibit the binding of IF3 to the 30S ribosomal subunits. On the other hand, IF3 bound to the 30S subunits was expelled by paromomycin-induced subunit association (70S formation). These results indicate that the stabilization of 70S ribosomes by paromomycin may in part be responsible for its inhibitory effects on translocation and ribosome recycling.

Background: Osteosarcoma (OS) represents the most common primary bone tumor in humans and in companion dogs, being practically phenotypically identical. There is a need for effective treatments to extend the survival of patients with OS. Here, we examine the dosimetry in beagle dogs and cross-reactivity with human tissues of a novel human antibody, IF3, that targets the insulin growth factor receptor type 2 (IGF2R), which is overexpressed on OS cells, making it a candidate for radioimmunotherapy of OS. Methods: [89Zr]Zr-DFO-IF3 was injected into three healthy beagle dogs. PET/CT was conducted at 4, 24, 48, and 72 h. RAPID analysis was used to determine the dosimetry of [177Lu]Lu-CHXA”-IF3 for a clinical trial in companion dogs with OS. IF3 antibody was biotinylated, and a multitude of human tissues were assessed with immunohistochemistry. Results: PET/CT revealed that only the liver, bone marrow, and adrenal glands had high uptake. Clearance was initially through renal and hepatobiliary excretion in the first 72 h followed by primarily physical decay. RAPID analysis showed bone marrow to be the dose-limiting organ with a therapeutic range for 177Lu calculated to be 0.487–0.583 GBq. Immunohistochemistry demonstrated the absence of IGF2R expression on the surface of healthy human cells, thus suggesting that radioimmunotherapy with [177Lu]Lu-CHXA”-IF3 will be well tolerated. Conclusions: Image-based dosimetry has defined a safe therapeutic range for canine clinical trials, while immunohistochemistry has suggested that the antibody will not cross-react with healthy human tissues.

ABSTRACT The oxazolidinones represent a new class of antimicrobial agents which are active against multidrug-resistant staphylococci, streptococci, and enterococci. Previous studies have demonstrated that oxazolidinones inhibit bacterial translation in vitro at a step preceding elongation but after the charging ofN-formylmethionine to the initiator tRNA molecule. The event that occurs between these two steps is termed initiation. Initiation of protein synthesis requires the simultaneous presence of N-formylmethionine-tRNA, the 30S ribosomal subunit, mRNA, GTP, and the initiation factors IF1, IF2, and IF3. An initiation complex assay measuring the binding of [3H]N-formylmethionyl-tRNA to ribosomes in response to mRNA binding was used in order to investigate the mechanism of oxazolidinone action. Linezolid inhibited initiation complex formation with either the 30S or the 70S ribosomal subunits fromEscherichia coli. In addition, complex formation withStaphylococcus aureus 70S tight-couple ribosomes was inhibited by linezolid. Linezolid did not inhibit the independent binding of either mRNA or N-formylmethionyl-tRNA toE. coli 30S ribosomal subunits, nor did it prevent the formation of the IF2–N-formylmethionyl-tRNA binary complex. The results demonstrate that oxazolidinones inhibit the formation of the initiation complex in bacterial translation systems by preventing formation of theN-formylmethionyl-tRNA–ribosome–mRNA ternary complex.

We showed previously that the sequence and (or) structural elements important for specifying the many distinctive properties of Escherichia coli initiator tRNA are clustered in the acceptor stem and in the anticodon stem and loop. This paper briefly describes this and reviews the results of some recently published studies on the mutant initiator tRNAs generated during this work. First, we have studied the effect of overproduction of methionyl-tRNA transformylase (MTF) and initiation factors IF2 and IF3 on activity of mutant initiator tRNAs mat are defective at specific steps in the initiation pathway. Overproduction of MTF rescued specifically the activity of mutant tRNAs defective in formylation but not mutants defective in binding to the P site. Overproduction of IF2 increased me activity of all mutant tRNAs having the CUA anticodon but not of mutant tRNA having me GAC anticodon. Overproduction of IF3 had no effect on the activity of any of me mutant tRNAs tested. Second, for functional studies of mutant initiator tRNA in vivo, we used a CAU→CUA anticodon sequence mutant mat can initiate protein synthesis from UAG instead of AUG. In contrast with me wild-type initiator tRNA, the mutant initiator tRNA has a 2-methylthio-N6-isopentenyl adenosine (ms2i6A) base modification next to the anticodon. Interestingly, this base modification is now important for activity of the mutant tRNA in initiation. In a miaA strain of E. coli deficient in biosynthesis of ms2i6A, the mutant initiator tRNA is much less active in initiation. The defect is specifically in binding to the ribosomal P site.Key words: initiator tRNA, initiation Factors, formylation, P site binding, base modification.

One of the bottlenecks in relation to the production of forest seedlings is irrigation, especially in less-tech nurseries. The small volumes of substrate, where the seedlings develop, lead to low water storage. This fact can lead to water deficit and significant losses in the production of seedlings, generating even mortality. The objective of this study was to determine the best irrigation frequencies for Eucalyptus grandis seedling production in less-tech nurseries. The experiment was conducted between 2013/08/04 and 2013/23/07 (106 days) and conducted in Chapadão do Sul county, Brazil. The experiment was set up in a randomized complete block design, with five replications. Four irrigation frequencies were tested: IF1 (one daily irrigation - 11:00 a.m.), IF2 (two daily irrigations - 11:00 a.m. and 7:00 p.m.), IF3 (three daily irrigations - 07:00 a.m., 11:00 a.m. and 7:00 p.m.) and IF4 (four daily irrigations - 07:00 a.m., 11:00 a.m., 3:00 p.m. and 7:00 p.m.). The sample units were composed of eight seedlings in 50 cm3 conical tubes filled with soil and vermiculite in a volume ratio of 1:1. The irrigation depth was estimated by reference evapotranspiration (Penman-Monteith) multiplied by a crop coefficient equal to two. Plant height, number of definitive leaves, shoot dry mass (root and total), seed quality index, survival and efficiency of water use by eucalyptus seedlings were evaluated. The average daily irrigation depth in the experimental period was 5.2 mm. Based on the results, it is recommended for eucalyptus seedling producers, in less-tech nursery, irrigation management twice per days. (11:00 a.m. and 7:00 p.m.).

This research investigates the impacts of blended treated wastewater (TWW) reuse with freshwater (FW) and irrigation frequency on corn (Zea Maize L.) crop yield and NPK (nitrogen, phosphorus, and potassium) soil content. The experiment comprised of three irrigation frequencies IF1: daily, IF2: every other day, and IF3: every 3 days; and five blended water treatments T100(100%TWW), T75(75%TWW and 25%FW), T50(50%TWW and 50%FW), T25(25%TWW and 75%FW), and T0(100%FW), in four replications. Results indicate that the significant effect of the irrigation frequency was mainly on corn cobs yield and consequently crop yield. Crop yield increases as the ratio of TWW increased in the blended irrigation water, with the highest significant yield (58,036 kg/ha) by using pure TWW(T100) and the lowest yield (37,695 kg/ha) was obtained by using FW (T0). Regardless of the irrigation frequency, the highest soil NPK content was obtained by using pure TWW (T100), while the lowest NPK soil content was obtained by using FW treatment (T0). Available soil N, P, and K contents in T100 treatment were significantly higher than that in T0 treatment by 50.4%, 62%, and 53%, respectively. Thus, the use of TWW in agricultural irrigation could provide a good balance of plant nutrients which can markedly increase crop yield and reduced the need for expensive commercial fertilizers.

Abstract Mean amplitudes of vibration of IF3 have been calculated from vibrational spectroscopic data in the temperature range between 0 and 1000 K. Bond properties of the molecule are dis­ cussed on the basis of these results. Some comparison with relat­ed species are made.

A number of model mDNAs and mRNAs, about 19 nucleotides in length, were obtained by automated synthesis and T7 RNA polymerase directed transcription from synthetic DNA templates, respectively. These mDNAs and mRNAs had Shine-Dalgarno sequences that were four to eight nucleotides long and their sequence was similar to the R17 coat protein initiation site. The effect of S1 or initiation factors (IFs) on the rate and extent of ribosomal binding of these mDNAs and mRNAs was investigated by nitrocellulose filtration. No effect was detected, suggesting that the main function of S1 or IFs included neither the direct recognition or binding of short messengers to the 30S subunits, nor their rejection as "false" for possessing the wrong sugars or an insufficient length. A pulse–chase experiment with one of the mRNAs also showed that S1 or IF3 did not influence the exchange rate of mRNA bound to the 30S subunit.Key words: ribosome, initiation, mRNA, ribosomal protein S1, initiation factor IF3.

Abstract In bacteria, the assembly factors tightly orchestrate the maturation of ribosomes whose competency for protein synthesis is validated by translation machinery at various stages of translation cycle. However, what transpires to the quality control measures when the ribosomes are produced with assembly defects remains enigmatic. In Escherichia coli, we show that 30S ribosomes that harbour assembly defects due to the lack of assembly factors such as RbfA and KsgA display suboptimal initiation codon recognition and bypass the critical codon–anticodon proofreading steps during translation initiation. These premature ribosomes on entering the translation cycle compromise the fidelity of decoding that gives rise to errors during initiation and elongation. We show that the assembly defects compromise the binding of initiation factor 3 (IF3), which in turn appears to license the rapid transition of 30S (pre) initiation complex to 70S initiation complex by tempering the validation of codon–anticodon interaction during translation initiation. This suggests that the premature ribosomes harbouring the assembly defects subvert the IF3 mediated proofreading of cognate initiation codon to enter the translation cycle.

ABSTRACT By primer extension inhibition assays, 70S ribosomes bound with higher affinity, or stability, than did 30S subunits to leaderless mRNAs containing AUG or GUG start codons. Addition of translation initiation factors affected ribosome binding to leaderless mRNAs. Our results suggest that translation of leaderless mRNAs might initiate through a pathway involving 70S ribosomes or 30S subunits lacking IF3.

The processes of association and dissociation of ribosomal subunits are of great importance for the protein biosynthesis. The mechanistic details of these processes, however, are not well known. In bacteria, upon translation termination, the ribosome dissociates into subunits which is necessary for its further involvement into new initiation step. The dissociated state of the ribosome is maintained by initiation factor 3 (IF3) which binds to free small subunits and prevents their premature association with large subunits. In this work, we have exchanged IF3 in Escherichia coli cells by its ortholog from Saccharomyces cerevisiae mitochondria (Aim23p) and showed that yeast protein cannot functionally substitute the bacterial one and is even slightly toxic for bacterial cells. Our in vitro experiments have demonstrated that Aim23p does not split E. coli ribosomes into subunits. Instead, it fixes a state of ribosomes characterized by sedimentation coefficient about 60S which is not a stable structure but rather reflects a shift of dynamic equilibrium between associated and dissociated states of the ribosome. Mitochondria-specific terminal extensions of Aim23p are necessary for “60S state” formation, and molecular modeling results point out that these extensions might stabilize the position of the protein on the bacterial ribosome.

, Penelitian ini merupakan penelitian deskriptif, dengan metode penelitian deskriptif kualitatif dengan menggunakan prosedur pengumpulan data menggunakan teknik observasi, wawancara dan dokumentasi. yang digunakan menggunakan trianggulasi data, sumber, teknik dan waktu. Untuk mendapatkan hasil penelitian yang sesuai penulis melakukan wawancara dengan beberapa informan yakni Saudara Jerry Anggoro, S.S selaku ketua IF3 DKI Jakarta, Raihan Madani selaku Atlet freestyle soccer dan juga Drs.H. Muhammad Sarkawi selaku ketua KORMI (Komite Olahraga Rekreasi dan Masyarakat Indonesia) Penelitian ini bertujuan untuk mengambangkan strategi pengembangan olahraga rekreasi freestyle soccer pada KORMI DKI Jakarta sehingga olahraga rekreasi freestyle soccer dapat berkembang dan semakin meluas dikalangan masyarakat. Freestyle soccer merupakan bagian dari seni memainkan bola yang dilakukan dengan sebuah gaya dan trik yang memilik unsur keindahan dan unsur rekreasi untuk memainkan olahrga ini. Hasil penelitian menunjukkan bahwa organisasi IF3 DKI Jakarta Belum bersinergi dengan baik bersama KORMI DKI Jakarta Sehingga tidak berjalannya sistem organisasi yang mengakibatkan tidak berkembang nya olahraga rekreasi freestyle soccer di DKI Jakarta. Setelah melakukan observasi peneltian ini yang harus dilakukan adalah membenahi sistem yang ada di organisasi dan memperbaiki kekurangan organisasi dengan adanya observasi ini agar tujuan di dalamnya dapat tercapai sehingga olahraga rekreasi freestyle soccer dapat berkembang.

ABSTRACT We demonstrate that the transfer of fully charged aminoacyl-tRNAs into peptides directed by the MS2 RNA template requires both ATP and GTP, initiation factors (IF1, IF2, and IF3), elongation factors (EF-Tu, EF-Ts, and EF-G), and the ribosomal ATPase (RbbA). The nonhydrolyzable analogue AMPPCP inhibits the reactions, suggesting that hydrolysis of ATP is required for synthesis. The RbbA protein occurs bound to ribosomes and stimulates the ATPase activity of Escherichia coli 70S and 30S particles. The gene encoding RbbA harbors four ATP binding domains; the C-terminal half of the protein bears extensive sequence similarity to EF-3, a ribosome-dependent ATPase. Here, we show that the antibiotic hygromycin B selectively inhibits the ATPase activity of RbbA. Other antibiotics with similar effects on miscoding, streptomycin and neomycin, as well as antibiotics that impair peptide bond synthesis and translocation, had little effect on the ATPase activity of RbbA on 70S ribosomes. Immunoblot analysis indicates that at physiological concentrations, hygromycin B selectively releases RbbA from 70S ribosomes. Hygromycin B protects G1494 and A1408 in the decoding region, and RbbA enhances the reactivity of A889 and G890 of the 16S rRNA switch helix region. Cross-linking and X-ray diffraction data have revealed that this helix switch and the decoding region are in close proximity. Mutations in the switch helix (889-890) region affect translational fidelity and translocation. The binding site of hygromycin B and its known dual effect on the fidelity of decoding and translocation suggest a model for the action of this drug on ribosomes.

In prokaryotic systems, the initiation phase of protein synthesis is governed by the presence of initiation factors that guide the transition of the small ribosomal subunit (30S) from an unlocked preinitiation complex (30S preIC) to a locked initiation complex (30SIC) upon the formation of a correct codon–anticodon interaction in the peptidyl (P) site. Biochemical and structural characterization of GE81112, a translational inhibitor specific for the initiation phase, indicates that the main mechanism of action of this antibiotic is to prevent P-site decoding by stabilizing the anticodon stem loop of the initiator tRNA in a distorted conformation. This distortion stalls initiation in the unlocked 30S preIC state characterized by tighter IF3 binding and a reduced association rate for the 50S subunit. At the structural level we observe that in the presence of GE81112 the h44/h45/h24a interface, which is part of the IF3 binding site and forms ribosomal intersubunit bridges, preferentially adopts a disengaged conformation. Accordingly, the findings reveal that the dynamic equilibrium between the disengaged and engaged conformations of the h44/h45/h24a interface regulates the progression of protein synthesis, acting as a molecular switch that senses and couples the 30S P-site decoding step of translation initiation to the transition from an unlocked preIC to a locked 30SIC state.

AbstractIrrigation water scheduling is crucial to make the most efficient use of ever-decreasing water. As excessive irrigation decreases yield, while imprecise application also causes various environmental issues. Therefore, efficient management of irrigation frequency and irrigation level is necessary to sustain productivity under limited water conditions. The objective of the current study is to assess the water productivity at various irrigation regimes during peanut crop growing seasons (2014 and 2015) in Eastern Mediterranean, Turkey. The field experiments were conducted with treatments consisting of three irrigation frequencies (IF) (IF1: 25 mm; IF2: 50 mm; and IF3: 75 mm of cumulative pan evaporation (CPE)), and four irrigation water levels (WL1 = 0.50, WL2 = 0.75, WL3 = 1.0, and WL4 = 1.25). WL1, WL2, WL3, and WL4 treatments received 50, 75, 100, and 125 of cumulative pan evaporation. The CSM-CROPGRO-Peanut model was calibrated with experimental data in 2014 and evaluated with second-year experimental data (2015). The model simulated seed yield and final biomass (dry matter) reasonably well with low normalized root mean square error (RMSEn) in various irrigation intervals. The model simulated reasonably well for days to anthesis (RMSE = 2.53, d-stat = 0.96, and r2 = 0.90), days to physiological maturity (RMSE = 2.55), seed yield (RMSE = 1504), and tops biomass dry weight at maturity (RMSE = 3716). Simulation results indicated good agreement between measured and simulated soil water content (SWC) with low RMSEn values (4.0 to 16.8% in 2014 and 4.3 to 18.2% in 2015). Further results showed that IF2I125 irrigation regime produced the highest seed yield. Generally, model evaluation performed reasonably well for all studied parameters with both years’ experimental data. Results also showed that the crop model would be a precision agriculture tool for the extrapolation of the allocation of irrigation water resources and decision management under current and future climate.

Abstract The objective of this study was to compare serological responses in low-risk calves vaccinated with various IBR-BVD-PI3-BRSV-Mannheimia haemolytica toxoid vaccines. Bovine herpes virus-1 (BHV-1) and M. haemolytica (MH) leukotoxin titers were analyzed to investigate any potential BHV-1 antigen interference to MH leukotoxin levels when administering multi-valent vaccines. Serology responses of BVD, BRSV, and PI3 were not within the scope of this study. Before enrolling in the study, weaned calves born to dams at two University of Arkansas beef cattle research stations that were not previously vaccinated with MH toxoid or BHV-1 were bled and sera harvested. Crossbred steer and heifer calves [n = 364; steers = 168, heifers = 196; body weight (BW) = 187.9 ± 2.8 kg] were used for this study at the two locations. Calves were stratified by MH titers, BHV-1 titers, and sex, and allotted to 1 of 5 treatments: 1) “BS GOLD” = IBR-BVD-PI3-BRSV-MH (Bovi-Shield GOLD One Shot, 2 mL/SQ; Zoetis); 2) “P5 + PRE” = IBR-BVD-PI3-BRSV-MH (Pyramid 5 + Presponse SQ, 2 mL/SQ; Boehringer Ingelheim Animal Health); 3) “BV Once” = IBR-BVD-PI3-BRSV-MH-PM (Bovilis Vista Once SQ, 2 mL/SQ; Merck Animal Health); 4) “IF3 + One Shot” = IBR-BVD-PI3-BRSV (Inforce 3, 2 mL/intranasal; Zoetis) and BVD-MH (One Shot BVD, 2 mL/SQ; Zoetis); and 5) “BN + BV” = IBR-BVD-PI3-BRSV (Bovilis Nasalgen 3-PMH, 2 mL/intranasal; Merck Animal Health) and BVD (Bovilis Vista BVD CFP, 2 mL/SQ; Merck Animal Health). Cattle were weighed on d 0, 7, 14, and 28 and blood samples were taken for MH leukotoxin titers. Serum samples were collected on d 0 and 28 for BHV-1 titers. For statistical analyses, data were analyzed using the PROC MIXED procedure of SAS. Statistical significance was determined at P ≤ 0.05, with tendencies at 0.05 &amp;lt; P ≤ 0.1. There was an overall treatment effect for MH serum antibody titers with BV Once and BN + BV vaccinated calves exhibiting decreased MH serum titers compared with calves vaccinated with BS GOLD, P5 + PRE, and IF3 + One Shot (P = 0.0457). There was a treatment by day interaction for BHV-1 titers (P = 0.0336). On d 28, BHV-1 serum titers were greatest in BS GOLD and BN + BV vaccinated calves, least in calves that received IF3 + One Shot vaccines, and intermediate for BV Once vaccinated calves. Morbidity and average daily gain did not differ between treatments (P &amp;gt; 0.27) throughout the 28-d study period.

The COVID-19 pandemic has impacted all aspects of life, including teaching and learning activities. Therefore, the entire community must be ready to do online learning, which will have different processes and results from normal learning. This study aims to determine the level of student satisfaction in online learning during the pandemic. This study uses attributes on Webqual 4.0 and processed using the integration of IPA and Kano. Based on the results of research conducted on 194 respondents, it was found that attributes US1 and IT4 were in the Beginning jewelry category, IT7 attributes were in the Defenseless Strategy point category, IT2 attributes were in the Fatal category, attributes IF1, IF2, IT3 was in the Major weapon category, attributes IF4, IF5 are included in the precious treasure category, and other attributes are included in the removed category because they have the Kano Indifferent category.

Unicellular tubes are components of internal organs and capillaries. It is unclear how they meet the architectural challenge to extend a centered intracellular lumen of uniform diameter. In an RNAi-based Caenorhabditis elegans screen, we identified three intermediate filaments (IFs)—IFA-4, IFB-1, and IFC-2—as interactors of the lumenal membrane-actin linker ERM-1 in excretory-canal tubulogenesis. We find that IFs, generally thought to affect morphogenesis indirectly by maintaining tissue integrity, directly promote lumenogenesis in this capillary-like single-cell tube. We show that ERM-1, ACT-5/actin, and TBB-2/tubulin recruit membrane-forming endosomal and flux-promoting canalicular vesicles to the lumen, whereas IFs, themselves recruited to the lumen by ERM-1 and TBB-2, restrain lateral vesicle access. IFs thereby prevent cystogenesis, equilibrate the lumen diameter, and promote lumen forward extension. Genetic and imaging analyses suggest that IFB-1/IFA-4 and IFB-1/IFC-2 polymers form a perilumenal triple IF lattice, sandwiched between actin and helical tubulin. Our findings characterize a novel mechanism of capillary-like lumenogenesis, where a tensile trilayered cytoskeletal endotube transforms concentric into directional growth.

ABSTRACT The crystal structure of a small, basic DNA binding protein, Sso10b2, from the thermoacidophilic archaeon Sulfolobus solfataricus was determined by the Zn multiwavelength anomalous diffraction method and refined to 1.85 Å resolution. The 89-amino-acid protein adopts a βαβαββ topology. The structure is similar to that of Sso10b1 (also called Alba) from the same organism. However, Sso10b2 contains an arginine-rich loop RDRRR motif, which may play an important role in nucleic acid binding. There are two independent Sso10b2 proteins in the asymmetric unit, and a plausible stable dimer could be deduced from the crystal structure. Topology comparison revealed that Sso10b2 is similar to several RNA-binding proteins, including IF3-C, YhhP, and DNase I. Models of the Sso10b2 dimer bound to either B-DNA or A-DNA have been constructed.