scholarly journals Genome Features of a New Double-Stranded RNA Helper Virus (LBCbarr) from Wine Torulaspora delbrueckii Killer Strains

2021 ◽  
Vol 22 (24) ◽  
pp. 13492
Author(s):  
Manuel Ramírez ◽  
Rocío Velázquez ◽  
Antonio López-Piñeiro ◽  
Alberto Martínez
Keyword(s):  
High Throughput Sequencing ◽  
Polymerase Activity ◽  
Helper Virus ◽  
Amino Acid Sequences ◽  
Dsrna Virus ◽  
Medium Size ◽  
Torulaspora Delbrueckii ◽  
Killer Strain ◽  
Large Size ◽  
Helper Activity

The killer phenotype of Torulaspora delbrueckii (Td) and Saccharomyces cerevisiae (Sc) is encoded in the genome of medium-size dsRNA viruses (V-M). Killer strains also contain a helper large size (4.6 kb) dsRNA virus (V-LA) which is required for maintenance and replication of V-M. Another large-size (4.6 kb) dsRNA virus (V-LBC), without known helper activity to date, may join V-LA and V-M in the same yeast. T. delbrueckii Kbarr1 killer strain contains the killer virus Mbarr1 in addition to two L viruses, TdV-LAbarr1 and TdV-LBCbarr1. In contrast, the T. delbrueckii Kbarr2 killer strain contains two M killer viruses (Mbarr1 and M1) and a LBC virus (TdV-LBCbarr2), which has helper capability to maintain both M viruses. The genomes of TdV-LBCbarr1 and TdV-LBCbarr2 were characterized by high-throughput sequencing (HTS). Both RNA genomes share sequence identity and similar organization with their ScV-LBC counterparts. They contain all conserved motifs required for translation, packaging, and replication of viral RNA. Their Gag-Pol amino-acid sequences also contain the features required for cap-snatching and RNA polymerase activity. However, some of these motifs and features are similar to those of LA viruses, which may explain that at least TdV-LBCbarr2 has a helper ability to maintain M killer viruses. Newly sequenced ScV-LBC genomes contained the same motifs and features previously found in LBC viruses, with the same genome location and secondary structure. Sequence comparison showed that LBC viruses belong to two clusters related to each species of yeast. No evidence for associated co-evolution of specific LBC with specific M virus was found. The presence of the same M1 virus in S. cerevisiae and T. delbrueckii raises the possibility of cross-species transmission of M viruses.

scholarly journals Genome Organization of a New Double-Stranded RNA LA Helper Virus From Wine Torulaspora delbrueckii Killer Yeast as Compared With Its Saccharomyces Counterparts

2020 ◽  
Vol 11 ◽  
Author(s):  
Manuel Ramírez ◽  
Rocío Velázquez ◽  
Matilde Maqueda ◽  
Alberto Martínez
Keyword(s):  
Genome Organization ◽  
Sequence Comparison ◽  
High Throughput Sequencing ◽  
Killer Toxin ◽  
Medium Size ◽  
High Sequence Identity ◽  
Torulaspora Delbrueckii ◽  
Killer Yeast ◽  
Dsrna Viruses ◽  
Extra Sequence

Wine killer yeasts such as killer strains of Torulaspora delbrueckii and Saccharomyces cerevisiae contain helper large-size (4.6 kb) dsRNA viruses (V-LA) required for the stable maintenance and replication of killer medium-size dsRNA viruses (V-M) which bear the genes that encode for the killer toxin. The genome of the new V-LA dsRNA from the T. delbrueckii Kbarr1 killer yeast (TdV-LAbarr1) was characterized by high-throughput sequencing (HTS). The canonical genome of TdV-LAbarr1 shares a high sequence identity and similar genome organization with its Saccharomyces counterparts. It contains all the known conserved motifs predicted to be necessary for virus translation, packaging, and replication. Similarly, the Gag-Pol amino-acid sequence of this virus contains all the features required for cap-snatching and RNA polymerase activity, as well as the expected regional variables previously found in other LA viruses. Sequence comparison showed that two main clusters (99.2–100% and 96.3–98.8% identity) include most LA viruses from Saccharomyces, with TdV-LAbarr1 being the most distant from all these viruses (61.5–62.5% identity). Viral co-evolution and cross transmission between different yeast species are discussed based on this sequence comparison. Additional 5′ and 3′ sequences were found in the TdV-LAbarr1 genome as well as in some newly sequenced V-LA genomes from S. cerevisiae. A stretch involving the 5′ extra sequence of TdV-LAbarr1 is identical to a homologous stretch close to the 5′ end of the canonical sequence of the same virus (self-identity). Our modeling suggests that these stretches can form single-strand stem loops, whose unpaired nucleotides could anneal to create an intramolecular kissing complex. Similar stem loops are also found in the 3′ extra sequence of the same virus as well as in the extra sequences of some LA viruses from S. cerevisiae. A possible origin of these extra sequences as well as their function in obviating ssRNA degradation and allowing RNA transcription and replication are discussed.

scholarly journals Relationships and Evolution of Double-Stranded RNA Totiviruses of Yeasts Inferred from Analysis of L-A-2 and L-BC Variants in Wine Yeast Strain Populations

2016 ◽  
Vol 83 (4) ◽  
Author(s):  
Nieves Rodríguez-Cousiño ◽  
Rosa Esteban
Keyword(s):  
Wine Yeast ◽  
Helper Virus ◽  
Medium Size ◽  
Coat Proteins ◽  
Double Stranded Rna ◽  
Content Type ◽  
Protein Toxin ◽  
Killer Toxins ◽  
Helper Activity ◽  
A Minor

ABSTRACT Saccharomyces cerevisiae killer strains secrete a protein toxin active on nonkiller strains of the same (or other) yeast species. Different killer toxins, K1, K2, K28, and Klus, have been described. Each toxin is encoded by a medium-size (1.5- to 2.3-kb) M double-stranded RNA (dsRNA) located in the cytoplasm. M dsRNAs require L-A helper virus for maintenance. L-A belongs to the Totiviridae family, and its dsRNA genome of 4.6 kb codes for the major capsid protein Gag and a minor Gag-Pol protein, which form the virions that separately encapsidate L-A or the M satellites. Different L-A variants exist in nature; on average, 24% of their nucleotides are different. Previously, we reported that L-A-lus was specifically associated with Mlus, suggesting coevolution, and proposed a role of the toxin-encoding M dsRNAs in the appearance of new L-A variants. Here we confirm this by analyzing the helper virus in K2 killer wine strains, which we named L-A-2. L-A-2 is required for M2 maintenance, and neither L-A nor L-A-lus shows helper activity for M2 in the same genetic background. This requirement is overcome when coat proteins are provided in large amounts by a vector or in ski mutants. The genome of another totivirus, L-BC, frequently accompanying L-A in the same cells shows a lower degree of variation than does L-A (about 10% of nucleotides are different). Although L-BC has no helper activity for M dsRNAs, distinct L-BC variants are associated with a particular killer strain. The so-called L-BC-lus (in Klus strains) and L-BC-2 (in K2 strains) are analyzed. IMPORTANCE Killer strains of S. cerevisiae secrete protein toxins that kill nonkiller yeasts. The “killer phenomenon” depends on two dsRNA viruses: L-A and M. M encodes the toxin, and L-A, the helper virus, provides the capsids for both viruses. Different killer toxins exist: K1, K2, K28, and Klus, encoded on different M viruses. Our data indicate that each M dsRNA depends on a specific helper virus; these helper viruses have nucleotide sequences that may be as much as 26% different, suggesting coevolution. In wine environments, K2 and Klus strains frequently coexist. We have previously characterized the association of Mlus and L-A-lus. Here we sequence and characterize L-A-2, the helper virus of M2, establishing the helper virus requirements of M2, which had not been completely elucidated. We also report the existence of two specific L-BC totiviruses in Klus and K2 strains with about 10% of their nucleotides different, suggesting different evolutionary histories from those of L-A viruses.

AISI TYPE S2

Alloy Digest ◽  
1982 ◽  
Vol 31 (12) ◽  
Keyword(s):  
Physical Properties ◽  
Tensile Properties ◽  
Tool Steel ◽  
Shock Loading ◽  
Heat Treating ◽  
Medium Size ◽  
Hard Case ◽  
Large Size ◽  
Steel Mills ◽  
Aisi Type

Abstract AISI Type S2 is a water-hardening tool steel with extreme toughness and resistance to shock loading. Even at a hardness of Rockwell C 59-60, it will bend before it breaks. When hardened in medium-size and large-size pieces, it acquires a hard case and a tough core. Sizes under 3/4-inch (19mm) diameter will water harden to the center. The extreme toughness of Type S2 makes it suitable for use in many applications where no other tool steel will hold up. Its many uses include chisels, rivet busters, spike mauls, screw drivers, punches and sledges. This datasheet provides information on composition, physical properties, hardness, elasticity, and tensile properties. It also includes information on forming, heat treating, machining, and joining. Filing Code: TS-408. Producer or source: Tool steel mills.

scholarly journals Exercise Intensity and Technical Involvement in U9 Team Handball: Effect of Game Format

2021 ◽  
Vol 18 (11) ◽  
pp. 5663
Author(s):  
Georgios Ermidis ◽  
Rasmus C. Ellegard ◽  
Vincenzo Rago ◽  
Morten B. Randers ◽  
Peter Krustrup ◽  
...  
Keyword(s):  
Heart Rate ◽  
Locomotor Activity ◽  
Exercise Intensity ◽  
High Intensity ◽  
High Speed ◽  
Medium Size ◽  
Total Distance ◽  
Team Handball ◽  
Large Size ◽  
Game Format

The purpose of this study was to quantify the exercise intensity and technical involvement of U9 boys’ and girls’ team handball during different game formats, and the differences between genders. Locomotor activity (total distance, distance in speed zones, accelerations, and decelerations), heart rate (HR), and technical involvement (shots, goals, and duels) metrics were collected during various 15 min game formats from a total of 57 Danish U9 players (37 boys and 20 girls). Game formats were a small size pitch (20 × 13 m) with 3 vs 3 players and offensive goalkeepers (S3 + 1) and 4 vs 4 players (S4), a medium size pitch (25.8 × 20 m) with 4 vs 4 (M4) and 5 vs 5 (M5) players, and a large size pitch (40 × 20 m) with 5 vs 5 (L5) players. Boys and girls covered a higher total distance (TD) of high-speed running (HSR) and sprinting during L5 games compared to all other game formats (p < 0.05; ES = (−0.9 to −2.1), (−1.4 to −2.8), and (−0.9 to −1.3) respectively). Players covered the highest amount of sprinting distance in L5 games compared to all other game formats (p < 0.01; ES = 0.8 to 1.4). In all the game formats, players spent from 3.04 to 5.96 min in 180–200 bpm and 0.03 min to 0.85 min in >200 bpm of the total 15 min. In addition, both genders had more shots in S3 + 1 than M5 (p < 0.01; ES = 1.0 (0.4;1.7)) and L5 (p < 0.01; ES = 1.1 (0.6;2.2)). Team handball matches have high heart rates, total distances covered, and high-intensity running distances for U9 boys and girls irrespective of the game format. Locomotor demands appeared to be even higher when playing on larger pitches, whereas the smaller pitch size and fewer players led to elevated technical involvement.

scholarly journals Heterogeneity of penicillin-non-susceptible group B streptococci isolated from a single patient in Germany

2019 ◽  
Vol 75 (2) ◽  
pp. 296-299 ◽  
Author(s):  
Mark van der Linden ◽  
Rafael Mamede ◽  
Natascha Levina ◽  
Peter Helwig ◽  
Pedro Vila-Cerqueira ◽  
...  
Keyword(s):  
High Throughput Sequencing ◽  
Genomic Analysis ◽  
Amino Acid Sequences ◽  
Capsular Type ◽  
Group B Streptococci ◽  
The Us ◽  
Type Ia ◽  
The Usa ◽  
Group B ◽  
Host Evolution

Abstract Objectives Streptococcus agalactiae [group B streptococci (GBS)] have been considered uniformly susceptible to penicillin. However, increasing reports from Asia and North America are documenting penicillin-non-susceptible GBS (PRGBS) with mutations in pbp genes. Here we report, to the best of our knowledge, the first two PRGBS isolates recovered in Europe (AC-13238-1 and AC-13238-2), isolated from the same patient. Methods Two different colony morphologies of GBS were noted from a surgical abscess drainage sample. Both were serotyped and antimicrobial susceptibility testing was performed by different methodologies. High-throughput sequencing was done to compare the isolates at the genomic level, to identify their capsular type and ST, to evaluate mutations in the pbp genes and to compare the isolates with the genomes of other PRGBS isolates sharing the same serotype and ST. Results Isolates AC-13238-1 and AC-13238-2 presented MICs above the EUCAST and CLSI breakpoints for penicillin susceptibility. Both shared the capsular type Ia operon and ST23. Genomic analysis uncovered differences between the two isolates in seven genes, including altered pbp genes. Deduced amino acid sequences revealed critical substitutions in PBP2X in both isolates. Comparison with serotype Ia clonal complex 23 PRGBS from the USA reinforced the similarity between AC-13238-1 and AC-13238-2, and their divergence from the US strains. Conclusions Our results support the in-host evolution of β-lactam-resistant GBS, with two PRGBS variants being isolated from one patient.

scholarly journals Hearing gain after myringoplasty in relation to the size of tympanic membrane perforation

2020 ◽  
Vol 25 (1) ◽  
pp. 54-59
Author(s):  
Sutanu Kumar Mondal ◽  
Ashim Kumar Biswas ◽  
Md Mahmudul Huq ◽  
Md Hasan Ali ◽  
Md Kamruzzaman ◽  
...  
Keyword(s):  
Hearing Loss ◽  
Tympanic Membrane ◽  
Bone Conduction ◽  
Tympanic Membrane Perforation ◽  
Medium Size ◽  
Hearing Gain ◽  
Bone Conduction Threshold ◽  
Large Size ◽  
Membrane Perforation ◽  
Air Conduction

Objectives: To assess hearing gain after successful myringoplasty in relation to the size of tympanic membrane perforation. Methods: This cross-sectional study was done in the department of otolaryngology and head neck surgery, BSMMU, Sahbag, Dhaka during the period of January 2009 to December 2010.A total of 60 patients were under went myringoplasty operation after taking detailed history, clinical examination and investigation. Preoperative and postoperative hearing assessment was done. Analysed data presented by various tables, graphics and figures. Results: In case of small size perforation preoperative mean bone conduction threshold was 7.66 dB, mean air conduction threshold was 34.14 dB and mean air bone gap was 26.48 dB. In case of medium size perforation preoperative mean bone conduction threshold was 9.61 dB, mean air conduction threshold was 44.48 dB. Mean air bone gap was 34.87 dB. In case of large size perforation preoperative mean bone conduction threshold was 13.12 dB, mean air conduction threshold was 59 dB, and mean air bone gap was 45.88 dB. Hearing loss increases with increasing size of perforation. Ahmed and Rahim (1979) showed in the study that hearing loss increases with increasing the size of the perforation which was relevant in the study. After myringoplasty post-operative mean air bone gap was 21.24 dB in small size, 21.74 dB in medium sized and 24 dB in large size. From the record improvement of mean air bone gap or hearing gain was 5.24 dB in small size perforation respectively. The different of air bone gap closure between small and medium size perforation was statistically significant by unpaired’ test. Conclusion: Hearing gain after myringoplasty is better in large size perforation. Bangladesh J Otorhinolaryngol; April 2019; 25(1): 54-59

Discovery and genome characterization of a new Nepovirus infecting grapevine

Plant Disease ◽  
2020 ◽  
Author(s):  
Maher Al Rwahnih ◽  
Olufemi Joseph Alabi ◽  
Min Sook Hwang ◽  
Tongyan Tian ◽  
Dimitre Mollov ◽  
...  
Keyword(s):  
High Throughput Sequencing ◽  
Phylogenetic Analyses ◽  
Virus Transmission ◽  
Amino Acid Sequences ◽  
Molecular Characteristics ◽  
Vitis Vinifera L ◽  
Sequencing Analysis ◽  
Biological Index ◽  
Chenopodium Amaranticolor ◽  
Sequence Contigs

In 2012, dormant canes of a proprietary wine grape (Vitis vinifera L.) accession were included in the collection of the University of California-Davis Foundation Plant Services. No virus-like symptoms were elicited when bud chips from propagated own-rooted canes of the accession were graft-inoculated onto a panel of biological index grape varieties. However, chlorotic ring symptoms were observed on sap inoculated Chenopodium amaranticolor Coste & A. Rein and C. quinoa Willd. plants, indicating the presence of a mechanically transmissible virus. Transmission electron microscopy of virus preps from symptomatic C. quinoa revealed spherical, non-enveloped virions of ~27 nm in diameter. And nepovirus-like haplotypes of sequence contigs were detected in both the source grape accession and recipient C. quinoa plants using high throughput sequencing analysis. A novel bipartite nepovirus-like genome was assembled from these contigs and the termini of each RNA segment were verified by RACE assays. The RNA1 (7,186-nt) of the virus encode a large polyprotein P1 of 231.1 kDa while the RNA2 (4,460-nt) also encode a large polyprotein P2 of 148.9 kDa. Each of the polyadenylated RNA segment is flanked by 5′- (RNA1=156-nt; RNA2=170-nt) and 3′- (RNA1=834-nt; RNA2=261-nt) untranslated region sequences that shared >90% identities between their corresponding sequences. Maximum-likelihood phylogenetic analyses of the conserved Pro-Pol amino acid sequences of Secoviridae species revealed the clustering of the new virus within the nepovirus clade. Considering its biological and molecular characteristics, and based on current criteria, we propose that the novel virus, named as grapevine nepovirus A (GNVA), be assigned as a member of the genus Nepovirus.

Vibration Transmissibility on Foot during Controlling and Operating Car Accelerator Pedal

2013 ◽  
Vol 471 ◽  
pp. 151-155
Author(s):  
A.R. Yusoff ◽  
Baba M. Deros ◽  
D.D.I Daruis
Keyword(s):  
Lower Extremity ◽  
Medium Size ◽  
Accelerator Pedal ◽  
Vehicle Engine ◽  
Large Size ◽  
Honestly Significant Difference ◽  
Significant Difference ◽  
Vibration Transmissibility ◽  
Post Hoc ◽  
The Relationship

The purpose of this paper is to discuss the vibration transmissibility from a cars body to pedal-pad, and the lower extremity, especially on the foot. The vibration transmissibility will occur when the foot interacts with the pedal-pad once the vehicle engine is turned on. The vibrations transmissibility will cause drivers discomfort, especially on the lower extremity due to fatigue and tiredness of the foot. The relationship between vibration transmissibility with the different sizes pedal-pad to the foot, and whether it can contribute to the foot discomfort during handling and controlling pedal-pad while driving is investigated. The variables are the different sizes of pedal-pad. Data was recorded from foot on three different sizes of pedal-pads. The data was measured in root mean square (r.m.s) unit, of the frequency weighted acceleration (m/s2) for every minute. The result for the foot-transmitted-vibration shows the ratio value of medium sized pedal-pad is greater than 1; compared to small sized and large sized pedal-pads which is less than 1. Analysis of One-way Anova shows that the mean of different sizes of pedal-pad is significant. Tukeyspost-hocwith HSD (Honestly Significant Difference) test on the foot transmitted-vibration proves that the means for medium size pedal-pad is the highest compared with the small and large size pedal-pads.

Diet, feeding behaviour and habitat utilisation of the blue stingray Dasyatis chrysonota (Smith, 1828) in South African waters

2003 ◽  
Vol 54 (8) ◽  
pp. 957 ◽  
Author(s):  
David A. Ebert ◽  
Paul D. Cowley
Keyword(s):  
Surf Zone ◽  
Stomach Contents ◽  
Size Class ◽  
Behaviour Pattern ◽  
Medium Size ◽  
Secondary Importance ◽  
Size Classes ◽  
Important Prey ◽  
Large Size ◽  
Large Size Class

Analysis of stomach contents for Dasyatis chrysonota revealed that diet varied with size and habitat. The diet of all size classes in the surf zone was comprised primarily of Callianassa spp., Donax spp. and unidentified polychaete species. The medium and large size classes fed primarily on Donax spp., whereas the very large size class fed mainly on Callianassa spp. Polychaetes were of secondary importance as prey for the medium size class. The diet of D. chrysonota in the nearshore zone consisted mainly of Balanoglossus capensis and Callianassa spp. Balanoglossus capensis decreased from an index of relative importance (IRI) of 75.3% for the medium size class to 59.9% for the very large size class, whereas Callianassa spp. increased from 22.8% to 39.4% between the medium and the very large size classes. The offshore zone was the only area in which small size class D. chrysonota were caught. The diet of these small D. chrysonota was primarily polychaetes and amphipods. Polychaetes increased in importance in the medium size class, but declined in each successively larger size class. Conversely, Pterygosquilla armata capensis became the single most important prey item for the very large size class, comprising an IRI of 50.9%. The behaviour pattern used by D. chrysonota to locate and extract prey is described.

scholarly journals Implementation of Lean Manufacturing in Saudi Manufacturing Organisations: An Empirical Study

2011 ◽  
Vol 339 ◽  
pp. 250-253 ◽  
Author(s):  
M. Azharul Karim ◽  
Mubarak Aljuhani ◽  
Ray Duplock ◽  
Prasad Yarlagadda
Keyword(s):  
Saudi Arabia ◽  
Lean Manufacturing ◽  
Organization Culture ◽  
Medium Size ◽  
Maintenance Optimization ◽  
Manufacturing Companies ◽  
Skilled Workers ◽  
Planning Systems ◽  
Large Size ◽  
Management Commitment

Abstract: In this paper, the level of lean manufacturing implementation by Saudi manufacturing companies is investigated, the extent of application of lean manufacturing practice is identified and the benefits and barriers of Lean implementation are evaluated. The results reported in this paper are based on data collected from a survey using a standard questionnaire administered to 120 manufacturers in Saudi Arabia. Evidence indicates that large size companies are more likely to implement and gain the advantages of lean manufacturing than small and medium size companies. The most implemented lean manufacturing tools are Computerized Planning Systems, TQM, Maintenance Optimization and CIP. Main barriers against lean manufacturing implementation include the organization culture, lack of management commitment and lack of skilled workers. Results also show that benefits gained from lean manufacturing implementation are significant and are correlated with the level of implementation of lean strategies.

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