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In December 2019, the Chinese city of Wuhan was the center of origin of a pneumonia-like disease outbreak with an unknown causative pathogen. The CDC, China, managed to track the source of infection to a novel coronavirus (2019-nCoV; SARS-CoV-2) that shares approximately 79.6% of its genome with SARS-CoV. The World Health Organization (WHO) initially declared COVID-19 as a Public Health Emergency of International Concern (PHEIC) and later characterized it as a global pandemic on March 11, 2020. Due to the novel nature of this virus, there is an urgent need for vaccines and therapeutics to control the spread of SARS-CoV-2 and its associated disease, COVID-19. Global efforts are underway to circumvent its further spread and treat COVID-19 patients through experimental vaccine formulations and therapeutic interventions, respectively. In the absence of any effective therapeutics, we have devised h bioinformatics-based approaches to accelerate global efforts in the fight against SARS-CoV-2 and to assist researchers in the initial phase of vaccine and therapeutics development. In this study, we have performed comprehensive meta-analyses and developed an integrative resource, "CoronaVR" (http://bioinfo.imtech.res.in/manojk/coronavr/). Predominantly, we identified potential epitope-based vaccine candidates, siRNA-based therapeutic regimens, and diagnostic primers. The resource is categorized into the main sections "Genomes," "Epitopes," "Therapeutics," and Primers." The genome section harbors different components, viz, genomes, a genome browser, phylogenetic analysis, codon usage, glycosylation sites, and structural analysis. Under the umbrella of epitopes, sub-divisions, namely cross-protective epitopes, B-cell (linear/discontinuous), T-cell (CD4+/CD8+), CTL, and MHC binders, are presented. The therapeutics section has different sub-sections like siRNA, miRNAs, and sgRNAs. Further, experimentally confirmed and designed diagnostic primers are earmarked in the primers section. Our study provided a set of shortlisted B-cell and T-cell (CD4+ and CD8+) epitopes that can be experimentally tested for their incorporation in vaccine formulations. The list of selected primers can be used in testing kits to identify SARS-CoV-2, while the recommended siRNAs, sgRNAs, and miRNAs can be used in therapeutic regimens. We foresee that this resource will help in advancing the research against coronaviruses. [ABSTRACT FROM AUTHOR]

The tendency to the formation of macrolide resistance in campylobacteriosis pathogens is considered as a serious threat to public health due to ubiquity of campylobacter strains resistant to a wide range of antibiotics, primarily fluoroquinolones and tetracyclines. To assess the prevalence of resistant Campylobacter spp., we performed screening for macrolide sensitivity among 40 Campylobacter jejuni strains isolated from raw milk, poultry product, and washings from the equipment of the poultry processing plants. Phenotypic resistance to erythromycin, the most popular antibiotic for the treatment of campylobacteriosis, was revealed in 27.5% C. jejuni strains; 10% strains were resistant to azithromycin. The search and selection for gene markers of Campylobacter resistance to macrolides was performed. It was found that the resistance of C. jejuni to erythromycin is realized mainly via synthesis of proteins that protect ribosomes (the presence of coding sequences was detected in 45% of the studied strains) and the transmembrane pump mechanism (efflux pump CmeABC genes were found in 36% isolates); both mechanisms are transmissible. Chromosomal mutations in the 23S rRNA sequence detected in 18% strains seem to play a less significant role. [ABSTRACT FROM AUTHOR]

The object of this paper is to highlight the literacy primer Paso a paso, by José Henriques Figueira, for its relevance to teaching of reading and writing. This book, little known in Spain, was very popular in Uruguay or Argentina at the end of the 19th century and the beginning of the 20th. The content and the form of this didactic material is analyzed to verify what are the pedagogical foundations on which it is based and to evaluate if they could somehow be transferred today to the school practice. [ABSTRACT FROM AUTHOR]

Copyright of Gaceta Médica de Caracas is the property of Academia Nacional de Medicina and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)

Aims: Diagnosis of leprosy, a chronic infection caused by Mycobacterium leprae, predominantly depends on clinical manifestations and histopathological analysis, hampering rapid and accurate diagnostics. Our aim was to increase accuracy of leprosy diagnosis by improving M. leprae's DNA detection based on polymerase chain reaction (PCR) technique using new specific primers for the RLEP repetitive sequence. Methods and Results: The specific target region, RLEP, of M. leprae's genome was selected based on comparative genomics. After confirming the specificity of this region, using blastn analysis, primers were designed and tested for their in silico specificity. To evaluate the specificity and sensitivity of these primers in vitro, 184 blood samples from patients were used in qPCR. The new primer pair LYON1/LYON2 produced 91% positive samples, whereas the current primer pair LP1/LP2 produced 46%. Specificity and DNA detection limit test were carried out to compare the efficiency of the developed primer pair. The LYON1/LYON2 primer showed 100% specificity, whereas LP1/LP2 showed 64%. The DNA detection limit of LYON1/LYON2 was 10 copies of bacterial genomes per millilitre, whereas LP1/LP2 was 1000 copies of bacterial genomes per millilitre. Conclusions: In conclusion, the developed LYON1/LYON2 primer pair presented to be a specific and sensitive new molecular marker for the diagnosis of leprosy. Significance and Impact of the Study: The development of a specific primer pair for the detection of the M. leprae genome through qPCR technique contributes to a fast, sensitive and specific diagnosis, which is essential to prevent spreading and progression of this disease. [ABSTRACT FROM AUTHOR]

Sex identification is crucial for behavioral, ecological and conservation studies. In monomorphic bird species, traditional methods for sex assessment require potentially invasive sampling and manipulation of individuals, such as through cloacal examination. Thus, molecular methods involving non-destructive sampling are needed to reduce the stress of animals under study. In monomorphic Adélie penguins (Pygoscelis adeliae), the use of a penguin-specific primer pair (PL/PR) to identify sex has been evaluated through DNA extracted from nondestructive feather samples. With penguin-specific primers for the CHD1 gene found on the sex chromosomes, we efficiently identified the sex of all sampled individuals (n = 92, belonging to 46 complete nesting pairs in study colonies; n = 26 belonging to surveyed individuals in a control colony). DNA extraction and PCR protocols to improve amplification success are reported. Our results were also confirmed through Sanger sequencing used to determine the previously unpublished sequences of the CHD1 alleles in Adélie penguins, which were subsequently deposited in GenBank and used to compare to similar species. In phylogenetic reconstructions, CHD1Z and CHD1W sequences of the Adélie Penguin grouped together with other Pygoscelis and were the sister group of Spheniscus and Eudyptes genera. [ABSTRACT FROM AUTHOR]

In sugarcane, fungal attack of Sporisorium scitamineum causing smut disease has resulted in noticeable yield losses and reduced cane quality. In the current study, RAPD technique was used for the identification of pathogen pathotypes involved in infection and development of whip smut disease in sugarcane. To study the genetic diversity in S. scitamineum from 12 sugarcane-producing areas of Sindh, 36 isolates of S. scitamineum were collected and assessed using random amplification of polymorphic DNA (RAPD) method. The genetic variation is amplified by PCR using six RAPD primers each of them consist of ten base pairs. Cluster analysis of the RAPD data was performed and S. scitamineum isolates were divided into six different clusters. The outcomes of our current studies will be helpful in the examination of the molecular diversity of S. scitamineum and for the development of the genetic foundation for the differentiation of pathogenicity and detecting the new races. Moreover, our outcomes might offer beneficial regarding pathogen selecting principle in smut resistant sugarcane germplasm and distribution of varieties. [ABSTRACT FROM AUTHOR]

Gene therapy promotes the expression of missing or defective genes and can be curative following administration of a single dose. Gene therapy is prohibited in equine athletes by regulatory bodies due to the high potential for abuse and novel analytical methods are needed for detection. The goal of this study was to detect the administration of an experimental gene therapy: a recombinant adeno‐associated viral vector (rAAV) carrying a transgene for the anti‐inflammatory cytokine IL‐10 (rAAV‐IL10). Twelve horses were randomly assigned to receive an intra‐articular injection of rAAV‐IL10 or phosphate buffered saline (vehicle) into a middle carpal joint. Plasma and synovial fluid were collected on days 0, 1, 2, 4, 7, 14, 28, 56, and 84. Primer pairs were designed to detect two unique regions of rAAV. Using quantitative real time PCR, both sets of primers detected rAAV for 14–28 days in joints and up to 4 days in plasma, in all six treated horses. In synovial fluid, rAAV was detected on day 56 in 4/6 horses by both primer sets, and on day 84 in 1/6 horses by one primer set. In plasma, rAAV was detected for 7 days in 5/6 horses, 14 days in 2/6 horses, and 28 days in 1/6 horses by one primer set, and was detected for up to 14 days in 1/6 horses by the other primer set. This study is the first to validate that quantitative real time PCR can be used to systemically detect the local administration of a gene therapy product to horses. [ABSTRACT FROM AUTHOR]

Root and collar rot disease caused by Phytophthora capsici (Leonian) is one of the most serious diseases in pepper, Capsicum annuum L. Knowledge about resistant genes is limited in pepper accessions to P. capsici. In this study, a diverse collection of 37 commercial edible and ornamental genotypes, and implication of seven novel candidate DEGs genes (XLOC_ 021757, XLOC_021821 , XLOC_012788 , XLOC_011295 , XLOC_021928 , XLOC_015473 and XLOC_000341) were up-regulated on resistant and susceptible pepper cultivars, through real-time polymerase chain reaction (qPCR) at transplanting and maturing stages. All seven related defense-gene candidates were up-regulated in all inoculated accessions to P. capsici , but these genes were highly expressed in resistant ones, 19OrnP-PBI, 37ChillP-Paleo, and 23CherryP-Orsh. The transcriptional levels of the seven related candidate DEGs were 5.90, 5.64, 5.62, 5.18, 3.94, 3.69, 3.16 folds higher in the resistant pepper genotypes, than the control ones, non-inoculated genotypes respectively. The candidate genes expressed herein, will provide a basis for further gene cloning and functional verification studies, and also will aid in an understanding of the regulatory mechanism of pepper resistance to P. capsici. • DEGs of XLOC group, are likely to be defense-related genes to P. capsici infection • and showed to what extend play important roles in combating the fungal pathogen, P. capsici , • As was in pepper cv. "19OrnP-PBI", which is regarded as one of the most resistant pepper over here. • These studies may be contributing to decrease the devastating effects of Phytophthora species, • On Solanaceae and other related family, through the producing program for resistant pepper varieties. [ABSTRACT FROM AUTHOR]

The study was conducted on the visitor preference for different modules of National Mango Database developed by ICAR-Central Institute for Subtropical Horticulture, Lucknow, India. The purpose behind creating the database was to develop a portal for providing information on different aspects related to mango. A number of modules were developed on the portal https://mangifera.res.in. Important modules of the database are on 566 phytochemicals, 294 protein sequences, 540 primers, 30,000 nucleotides, 1690 EST, 66 field gene banks holding >4500 accessions, nurseries, processing units, exporters, species and varieties. A module on online phenology monitoring system and a web tool to compare test variety with reference variety was also developed as per DUS guidelines for suggesting reference variety after comparison with candidate's variety. Agumented knowledge in the database is attracting high number of visitors from all over the world for collecting useful information available in different modules of the portal. The present analysis deals with the popularity of different modules through collated information at the portal. Digital products as an outcome may be developed for the use by farmers, traders, scientists, students and mango lovers. On the basis of analysis made for hits it can be said that mango database becoming popular among people as a unique mango repository and people around the world are showing interest in its different modules. [ABSTRACT FROM AUTHOR]

The genus Pelophylax has been considered a model organism for understanding hybridization. Despite being the most diverse within the Western Palearctic group of Pelophylax , the ridibundus / bedriagae lineage that includes six species, remains largely understudied, revealing many knowledge gaps in regards to their evolution and conservation. Using genetic data from populations among species inhabiting contact zones could prove vital in filling these gaps. We tested 17 microsatellite markers for cross-species amplification in mainland Pelophylax species distributed in southern Balkans and evaluated their power to successfully detect population/species structure. Importantly, we examined their potential for identifying hybrids and backcrosses between known hybridized species. We detected 12 highly polymorphic loci that cross-amplified all species that showed no significant Linkage Disequilibrium and were able to discriminate among species and between parental and hybrids. We suggest their future use in genetic studies for the genus Pelophylax in Greece, including the identification of contact zones. [ABSTRACT FROM AUTHOR]

The analysis of telomere dynamics in birds is a growing research field providing important findings on ecological and environmental variations in individuals' aging, fitness and lifespan. Real‐time quantitative PCR (qPCR) has gained much interest for the evaluation of telomere length in birds. However, the assessment of several key preanalytical and analytical factors to optimize the method for achieving reproducible results, and the influence of these factors on the conclusions of each study, have been generally overlooked. In this study, we assessed the performance of eight commercially‐available qPCR master mixes in the amplification of telomere fragments in two bird species (zebra finch Taeniopygia guttata and red‐billed chough Pyrrhocorax pyrrhocorax). We observed that qPCR master mixes influence the telomere primer binding to target sequences and the amplification specificity within and between bird species, although PCR amplification efficiencies were very close to 100% for all master mixes. These findings indicated that the suitability of the master mix and other analytical factors must be carefully evaluated before starting a telomere dynamics study, especially when the technique has not previously been used in the species. We also showed that optimal PCR amplification efficiencies do not translate to good qPCR telomere assays, and that inference about amplification specificity based only on melting curve data can lead to misleading conclusions. Overall, this work highlights the complexity of qPCR optimization for the study of telomere length in birds. [ABSTRACT FROM AUTHOR]

Apple market takes third place in the world after bananas and grapes. Apple trees are affected by viruses and virus-like diseases, which cause significant economic losses. The most common viruses in the apple tree are Apple chlorotic leafspot virus (ACLSV), Apple stem pitting virus (ASPV), and Apple stem grooving virus (ASGV). Tomato ringspot virus (ToRSV) and apple mosaic virus (ApMV) also cause significant economic losses. Timely detection of viruses and using virus-free planting materials will reduce crop losses. In this work, we performed genetic analysis of ACLSV, ASGV, ASPV, ApMV, ToRSV, and developed RT-PCR test-system for simultaneous detection of these viruses in apple tree materials. The sequences of the developed primers for the test-system are patented. The planting materials imported from Turkey and Italy were investigated for the presence of apple viruses. The most common virus in the samples is ACLSV (60%) followed by ASPV (34%), ASGV (30%) and ApMV (2%). ToRSV virus was not detected at all. Analysis of samples showed that 60% of planting materials were infected with at least one virus, some samples were infected with 3 viruses simultaneously. Mandatory certification of imported planting materials will reduce crop losses of apple orchards. [ABSTRACT FROM AUTHOR]

This study was carried out in the biotechnology laboratories of the Seed Inspection and Certification Department - Ministry of Agriculture during 2019, to study the relation among molecular five inbred lines with hybrid crosses of maize. to estimate the genetic diversity and determine the degree of genetic similarity among them using the ISSR based on PCR technic with 6 primers. All primers produced 79 bands at a rate of 13.2 bands per primer. The number of amplified bands ranged from 10 to the primer UBC 835 and 16 in the UBC 897, with a molecular length ranged between 100 - 1700 bp. The total number and percentage of polymorphic bands was 71 and 90% respectively. The primers UBC 856 and UBC 866 the highest percentage of had polymorphic bands of 100%. While the UBC 835 showed the lowest (70%). The highest efficiency and specificity were 12.7% and 9.9% respectively in UBC835. Based on molecular marker data and genetic similarity using the UPGMA method in establishing the dendrogram, the study showed that genetic similarity between the inbred lines ranged from 0.105 to 0.40, the highest (0.40) between the two strains BK121 and Zm6 , While the lowest degree of similarity (0.105) was between the two strains BK121 and ART-817 . [ABSTRACT FROM AUTHOR]

Copyright of Revista CIENCIA UNEMI is the property of Universidad Estatal de Milagro (UNEMI) and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)

Diet and health status directly or indirectly affect the gut microbiota. Mongolians have a varied intestinal Lactobacillus. However, how the diet regulates intestinal Lactobacillus and affects health has not been fully discussed. In this research, we engaged 72 adult Mongolian participants. Conducting a physical examination and Dietary assessments to understood their dietary habits and health status. Then, Designed a pair of Lactobacillus -specific primers to seeking correlations between their diet, health, and intestinal Lactobacillus. The results show that the Lactobacillus -specific primer (Lac16s) used was highly effective in amplifying Lactobacillus sequences. The amplified fragment effectively differentiated various Lactobacillus species. We deciphered the Lactobacillus diversity in Mongolian participants, discovering notable differences between the normal and overweight individuals. Specifically, the overweight group exhibited higher abundances of Ligilactobacillus salivarius , Limosilactobacillus mucosae , Ligilactobacillus ruminis , and Limosilactobacillus reuteri , whereas the normal weight group had a pronounced presence of Lacticaseibacillus paracasei. A deeper analysis established that vitamin intake profoundly influences the Lactobacillus composition in Mongolian intestines. Additionally, the health of the participants was closely tied to the presence of Lacticaseibacillus paracasei and Limosilactobacillus mucosae. Scrutinizing the relationships between Lactobacillus strains and metabolic function revealed a significant link between most metabolic pathways, such as ascorbate and aldarate metabolism, fructose and mannose metabolism, and histidine metabolism, especially with Lacticaseibacillus paracasei. This strain may be influenced by dietary habits. In conclusion, we infer that the Lactobacillus composition in Mongolian intestines is intertwined with their diet and health. [ABSTRACT FROM AUTHOR]

The reinvestigation of tetrazene single crystalline material by means of X-ray methods resulted in a slightly different structure when compared to previously published data. Reaction conditions responsible for different crystalline modification formation were investigated. Newly described C form was found to be the primary reaction product and the combined action of temperature and the presence of water over time is required for the transition to the A form. Both forms were described by X-ray powder diffraction. Tetrazene was also subjected to infrared and Raman spectroscopy, which allowed differentiating between the forms. The molecule was isotopically labeled with 15N atoms at two different locations (ring and nitrogen sidechain) and employed in assigning vibrational modes to the resulting bands. Differences between sensitivities to mechanical stimuli of the two modifications were investigated and found industrially insignificant. In the same vein, the performance of either modification in primer composition and primer was identical.

Identification has similarities and differences with other kinds of assessment activity: quality assessment, control management and certification. The final result of identification is verification of compliance or detection of falsification. Common features are tests for definition of actual values. This paper studies the design of universal primers for type identification of fruit raw material (strawberry, gooseberry, cherry, raspberry, banana, wild rose, kiwi). To further verify the specificity of primers, sequencing of fragments is produced, which are read by each from the primer pairs. For this purpose, 8 polymerase chain reactions (PCR-reactions) are initiated, one from each primer pair corresponding to one type of raw material. A single alignment matrix for each of the studied objects is created as a result. Re-verification of each matrix is conducted for the presence of read errors or other disputed single- nucleotide substitutions. It is stated that the alignment matrices of the nucleotide sequences of raspberry, strawberry (fragaria viridis), gooseberry, wild rose, cherry, banana and kiwi are aligned on all sides and the protruding "bases" do not disturb the future work of programmes for the primers design. Universal non-intersecting primers are chosen to identify the fruit raw material under studying. As a result of the use of various software packages and of the database GenBank NCBI, we managed to find a suitable DNA zone for each of the tested samples of fruit raw material at the level of generic differentiation for further development on its basis of the universal primers. It is zone 18S rDNA. All the found sequences have both the conservative part for planting a pair of primers, and the variable one for reliable identification of species or for phylogenetic analysis. As part of the study, all samples of fruit raw material have been identified

The objective of the study was to describe how much rat and swine primer developed from cytochrome b could detect rat and pork in processed beef products sold in North Maluku. The settings of the study were the traditional markets and supermarkets in several cities in North Maluku such as Ternate, Tidore Kepulauan, West Halmahera, North Halmahera, Central Halmahera, South Halmahera, East Halmahera, Sula Island and Morotai Island. The data collection lasted between May and June, 2015. The samples were analyzed in the Biotechnology Lab of Unkhair in July, 2015. To detect rat and swine DNA, the researchers used the PCR (Polymerase Chain Reaction) method with Top Taq master mix Kit kit (250) (Catalog no. 200403) Swine Primer: Forward: 5'CTA CAT AAG ATAT ATC CAC CAC A 3 'Reverse: 5' ACA TTG TGG GAT CTT CTA GGT 3 'Product size: 290 bp. Rat Primer: forward SIM (5'-GACCTCCCAGCTCCATCAAACATCTCATCTTGATGAAA-3'). Reverse (5'GAATGGGATTTT GTTGGAGTTT-3 '). Out of 41 samples, sample 3, 4 and 5 taken in Jailolo contained rat DNA (positive); the samples were amplified with 499 base pair length (bp). In addition, sample 2, 7, 8 and 10 from Ternate as well as sample 4 from Morotai Island was also found positive (containing rat DNA). In terms of swine DNA, all of the samples came back negative. The amplification showed that none of the meatball samples contained pork. No pig DNA was amplified in the gel.

Abstract Background The accurate tracking of Cryptosporidium in faecal, water and/or soil samples in water catchment areas is central to developing strategies to manage the potential risk of cryptosporidiosis transmission to humans. Various PCR assays are used for this purpose. Although some assays achieve specific amplification from Cryptosporidium DNA in animal faecal samples, some do not. Indeed, we have observed non-specificity of some oligonucleotide primers in the small subunit of nuclear ribosomal RNA gene (SSU), which has presented an obstacle to the identification and classification of Cryptosporidium species and genotypes (taxa) from faecal samples. Results Using a novel bioinformatic approach, we explored all available Cryptosporidium genome sequences for new and diagnostically-informative, multi-copy regions to specifically design oligonucleotide primers in the large subunit of nuclear ribosomal RNA gene (LSU) as a basis for an effective nested PCR-based sequencing method for the identification and/or classification of Cryptosporidium taxa. Conclusion This newly established PCR, which has high analytical specificity and sensitivity, is now in routine use in our laboratory, together with other assays developed by various colleagues. Although the present bioinformatic workflow used here was for the specific design of primers in nuclear DNA of Cryptosporidium, this approach should be broadly applicable to many other microorganisms.

The pattern of genetic diversity among 92 genotypes of soybean from 5 different origins/sources (Pakistan, the USA, Asian Vegetable Research Development Centre (AVRDC), Japan and North Korea) was analyzed using randomly amplified polymorphic DNA markers. Out of 20 random primers 6 tested, 10 were polymorphic among genotypes and they yielded 107 markers, with an average of 10.7 markers per primer. The proportion of polymorphic bands within genotypes ranged from 0.47 to 0.71 with an average of 0.59. Pakistani and US genotypes exhibited the highest number of polymorphic bands (95%), while North Korean genotypes revealed the lowest (60%). The mean band frequency of the primers among genotypes was 0.57 with a range of 0.08–0.99. The Shannon’s index and Nei’s genetic diversity index revealed that primer OPF-06 showed maximum genetic diversity among the genotypes. Dendrogram constructed using Unweighted Pair Group Mean Average (UPGMA) method divided the genotypes into 5 main groups consisting of 13 clusters. The results of cluster analysis indicated that the genetic diversity between Pakistani and US or AVRDC genotypes is much larger than that between Pakistani and North Korean or Japanese genotypes. The Pakistani genotypes had distinct bands from plant introductions. Therefore, the Pakistani genotypes may be useful to soybean breeders.

Description of the subject. This paper discusses the influence of amplicon length on real-time PCR results. Objectives. The aim of the experiments was to show that amplicon size has an influence on detection. Method. Tests were performed on genomic and plasmid DNA. Double-dye probes and SYBR® Green were used for detection by real-time PCR. Primers were selected in order to produce fragments with increasing sizes. Experiments dealt with two targets: an endogenous target for soybean (part of the lectin gene) and a transgenic target (junction P35S-CTP of the MON40-3-2 soybean). Results. The results show that the kinetics of amplification curves evolve as a function of amplicon length, and smaller amplicons yield a higher level of fluorescence for the plateau phase. DNA degradation within the sample as well as the principles of fluorescence acquisition as a function of the chemistry used can also be factors. Conclusions. It was experimentally shown that the observed effect is linked to the suboptimal elongation temperature used in real-time PCR. Detection using SYBR® Green is less impacted as the loss of efficiency is partially compensated by the greater integration of SYBR® Green molecules in the larger fragments.

Objective: Testing of methods to enhance the shear bond strength (SBS) between orthodontic metal brackets and amalgam by sandblasting and different primers. Materials And Methods: Three hundred samples of amalgam restorations (KerrAlloy®) were prepared in self-cured acrylic blocks, polished, and divided into two groups: nonsandblasted and sandblasted. Each group was divided into five subgroups with different primers used in surface treatment methods, with a control group of bonded brackets on human mandibular incisors. Following the surface treatments, mandibular incisor brackets (Unitek®) were bonded on the amalgam with adhesive resin (Transbond XT®). The SBS of the samples was tested. The adhesive remnant index (ARI) and failure modes were then determined under a stereo-microscope. Two-way analysis of variance, Chi-square, and Kruskal–Wallis tests were performed to calculate the correlations between and among the SBS and ARI values, the failure modes, and surface roughness results. Results: There were statistically significant differences of SBS among the different adhesive primers and sandblasting methods (P < 0.05). The sandblasted amalgam with Assure Plus® showed the highest SBS (P < 0.001). Samples mainly showed an ARI score = 1 and mix-mode failure. There was a statistically significant difference of surface roughness between nonsandblasted amalgam and sandblasted amalgam (P < 0.05), but no significant differences among priming agents (P > 0.05). Conclusions: Using adhesive primers with sandblasting together effectively enhances the SBS between orthodontic metal brackets and amalgam. The two primers with the ingredient methacryloxydecyl dihydrogen phosphate (MDP) monomer, Alloy Primer® and Assure Plus®, were the most effective. Including sandblasting in the treatment is essential to achieve the bonding strength required.

Ocimum tenuiflorum Linn. (Lamiaceae) is an aromatic plant with a lot of potential medicinally, industrially and domestically. There are different morpho-types of O. tenuiflorum, with different shades of purple pigment on their leaves. Very few studies have been done to study the genetic variation among O. tenuiflorum morpho-types. Thus, in the present study, four O. tenuiflorum morpho-types that varied in pigmentation were characterised using RAPD marker as a molecular tool. Eight different primers collectively amplified 64 bands in the four morpho-types analysed. The RAPD technique revealed that the varied amount of purple pigment in O. tenuiflorum morpho-types is not due to environmental factor alone. The UPGMA clustering algorithm based on RAPD data grouped the four morpho-types into two major groups, with O. tenuiflorum (T2) with very light purple leaves and stem morpho-type, more related to O. tenuiflorum (T3) that have deep purple leaves and stem. The result was able to show the phylogenetic relationships within the species. The low level of genetic diversity observed in O. tenuiflorum can be attributed to the mode of reproduction and the reproductive biology of the species, due to the fact that they are likely to be natural hybrids produced through cross pollination.

Background Sexual development in Flammulina velutipes is controlled by two different mating type loci (HD and PR). The HD locus contains homeodomain (Hd) genes on two separate HD subloci: HD-a and HD-b. While the functionality of the HD-b sublocus has been largely confirmed, the status and content of the HD-a sublocus has remained unclear. Methods To examine the function of the HD-a sublocus, genome sequences of a series of F. velutipes strains were analyzed and tested through series of amplification by specific primer sets. Furthermore, activity of di-allelic HD-a locus was confirmed by crossing strains with different combinations of HD-a and HD-b subloci. Results Sublocus HD-b contained a large variety of fixed Hd1/Hd2 gene pairs, while the HD-a sublocus either contained a conserved Hd2 gene or, a newly discovered Hd1 gene that was also conserved. Identification of whole HD loci, that is, the contents of HD-a and HD-b subloci in a strain, revealed that strains with similar HD-b subloci could still form normal dikaryons if the two genes at the HD-a sublocus differed. At least di-allelic HD-a sublocus, is thus indicated to be actively involved in mating type compatibility. Conclusions HD-a sublocus is active and di-allelic. Using the new information on the HD subloci, primers sets were developed that specifically amplify HD-a or HD-b subloci in the majority of F. velutipes strains. In this way, unknown HD mating types of F. velutipes can now be quickly identified, and HD mating type compatibility conferred by HD-a or HD-b can be confirmed by PCR.

Fluorescence in situ hybridization (FISH) and 16S rRNA gene amplicon sequencing are commonly used for microbial ecological analyses in biological enhanced phosphorus removal (EBPR) systems, the successful application of which was governed by the oligonucleotides used. We performed a systemic evaluation of commonly used probes/primers for known polyphosphate-accumulating organisms (PAOs) and glycogen-accumulating organisms (GAOs). Most FISH probes showed blind spots and covered nontarget bacterial groups. Ca . Competibacter probes showed promising coverage and specificity. Those for Ca . Accumulibacter are desirable in coverage but targeted out-group bacteria, including Ca . Competibacter, Thauera , Dechlorosoma , and some polyphosphate-accumulating Cyanobacteria. Defluviicoccus probes are good in specificity but poor in coverage. Probes targeting Tetrasphaera or Dechloromonas showed low coverage and specificity. Specifically, DEMEF455, Bet135, and Dech453 for Dechloromonas covered Ca . Accumulibacter. Special attentions are needed when using these probes to resolve the PAO/GAO phenotype of Dechloromonas . Most species-specific probes for Ca . Accumulibacter, Ca . Lutibacillus, Ca . Phosphoribacter, and Tetrasphaera are highly specific. Overall, 1.4% Ca . Accumulibacter, 9.6% Ca . Competibacter, 43.3% Defluviicoccus , and 54.0% Dechloromonas in the MiDAS database were not covered by existing FISH probes. Different 16S rRNA amplicon primer sets showed distinct coverage of known PAOs and GAOs. None of them covered all members. Overall, 520F-802R and 515F-926R showed the most balanced coverage. All primers showed extremely low coverage of Microlunatus (<36.0%), implying their probably overlooked roles in EBPR systems. A clear understanding of the strength and weaknesses of each probe and primer set is a premise for rational evaluation and interpretation of obtained community results., Competing Interests: The authors declare no competing interests., (© The Author(s) 2024. Published by Oxford University Press on behalf of the International Society for Microbial Ecology.)

Real-time RT-PCR for the detection of epizootic hemorrhagic disease virus (EHDV) in clinical samples is a fast and sensitive tool for the diagnosis and confirmation of disease. Several real-time RT-PCR methods have been reported over the last 10 years. In this chapter, we describe seven duplex real-time RT-PCR assays to amplify part of genome segment 2 of EHDV to enable serotype identification. The assay includes the detection of an endogenous control gene-beta-actin., (© 2024. The Author(s), under exclusive license to Springer Science+Business Media, LLC, part of Springer Nature.)

The emergence of EHDV in Europe during the autumn of 2022 reinforces the need for molecular tools (RT-PCR) for rapid detection of animals infected with this virus. Viral genome testing can be performed on whole blood under anticoagulant, spleen, and bloody organ homogenates from ruminants. It can also be performed on cell culture following viral isolation tests. Various so-called classical or end-point RT-PCRs will be described, which permit the amplification of a part of the viral genome (targeting segment 7) allowing the detection of EHDV whatever the serotype (pan-RT-PCR) and also to amplify a portion of the gene coding the viral protein (VP) 2 enabling serotyping. The PCR amplification products are visualized by agarose gel electrophoresis. Sequencing of the type-specific RT-PCR amplification products allows for the serotype of the virus to be determined., (© 2024. The Author(s), under exclusive license to Springer Science+Business Media, LLC, part of Springer Nature.)

The purpose of this article was to evaluate the performance of a poly(aniline‐co‐anisidine)/iron waste composite (COIW) as an anticorrosive pigment for primer coatings. A procedure was outlined to prepare COIW and evaluate its anticorrosive properties following the electrochemical behavior and corrosion test of the coated steel. Various zinc‐rich epoxy primers were formulated by replacing part of the zinc powder (ZP) pigment with COIW. In all formulas, the ratio of pigment to epoxy resin was fixed at 2. In the pigment mix, the percentage of COIW was ranged from 5% to 60% by weight to the ZP. The performance of the formulated primers was estimated by the salt immersion test and potentiodynamic polarization measurement (corrosion potential, polarization resistance, and corrosion rate essays). It was demonstrated that COIW is effective in protecting steel from corrosion when it is used in combination with ZP. [ABSTRACT FROM AUTHOR]

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Thermophilic microorganisms are of special interest for phylogenetics and research in prokaryotic evolution, since many of them belong to deep branches of the tree of life. For this reason a number of broadly used universal primer systems for the 16S rRNA gene eliminate certain groups of thermophilic prokaryotes from their detection spectra. In the present work, the known 16S rRNA gene primer systems were analyzed in order to determine their ability to reveal members of deep phylogenetic lineages containing thermophilic microorganisms. It was shown that application of most of the published primer systems could result in elimination of certain groups of thermophilic prokaryotes. In silico analysis of existing primer systems was used to select the primer system for the V3‒V4 region of the 16S rRNA gene, which minimized elimination of thermophilic prokaryotic groups. Comparison of the proposed system with the previously published ones was carried out using high-throughput sequencing. Statistical analysis of the sequencing results based on the Shannon and Chao1 indexes revealed high efficiency of the proposed system for analysis of microbial communities of Kamchatka hot springs. [ABSTRACT FROM AUTHOR]

Ocimum tenuiflorum Linn. (Lamiaceae) is an aromatic plant with a lot of potential medicinally, industrially and domestically. There are different morpho-types of O. tenuiflorum, with different shades of purple pigment on their leaves. Very few studies have been done to study the genetic variation among O. tenuiflorum morpho-types. Thus, in the present study, four O. tenuiflorum morpho-types that varied in pigmentation were characterised using RAPD marker as a molecular tool. Eight different primers collectively amplified 64 bands in the four morpho-types analysed. The RAPD technique revealed that the varied amount of purple pigment in O. tenuiflorum morpho-types is not due to environmental factor alone. The UPGMA clustering algorithm based on RAPD data grouped the four morpho-types into two major groups, with O. tenuiflorum (T2) with very light purple leaves and stem morpho-type, more related to O. tenuiflorum (T3) that have deep purple leaves and stem. The result was able to show the phylogenetic relationships within the species. The low level of genetic diversity observed in O. tenuiflorum can be attributed to the mode of reproduction and the reproductive biology of the species, due to the fact that they are likely to be natural hybrids produced through cross pollination. [ABSTRACT FROM AUTHOR]

Seventeen genotypes viz., CIM707, 4-F, RH1, NIAB111, MS40, MNH552, CIM443, BH118, CIM1100, CIM473, CIM482, NNH554, CIM448, CIM240, CIM435, NNH93 and NIAB78 of Gossypium hirsutum L. were collected from different cotton research organizations of Pakistan. The current study was conducted at Department of Plant Breeding and Genetics, University of Agriculture, Faisalabad during the year 2017. These varieties were grown and used for molecular studies using SSR and RAPD markers to amplify the genomic DNA. In SSR analysis, 45 primers were found polymorphic while in RAPD analysis only 14 primers found polymorphic. Maximum polymorphism was revealed by BNL and NAU primers whereas minimum polymorphism was revealed by JESPER and CIR primers. It was also found that no single DNA primer could deduct polymorphism among all genotypes specifically while several primers amplified polymorphic DNA fragments among few genotypes. It is obvious from similarity matrix that the genetic similarity among seventeen genotypes ranged from 37.21 to 86.05%. The highest similarity was found between CIM1100 and CIM 448 (86.05%) while NIAB111 and RH1 were most dissimilar (37.21%). The dendrogram proved that the majority of the genotypes were clustered into three groups. However, cotton genotypes i.e. CIM707, CIM240, CIM443, CIM473, CIM435 and CIM482 were clustered in group A; CIM 1100, CIM448, MNH 552, and MNH 554 were clustered in group B, whereas NIAB 78 and MNH93 in group C. The four genotypes viz: MS40, 4F, NIAB111 and RH1 remained ungrouped. They do not form any cluster with other genotypes and maintained their individuality. This highlights that mostly cultivated species has relatively narrow genetic base might be due to employing similar parents for the development of new cultivars at a particular breeding institute. Thus there is a prime need for expending the genetic base of most commonly cultivated cotton through hybridization among less commonly cultivated species. [ABSTRACT FROM AUTHOR]

aim,1 Laith AL-Eitan,2 Nour Abdo,3 Asem Alqudah,4 Hatem Aman11Department of Biotechnology and Genetic Engineering, Jordan University of Science and Technology, Irbid 22110, Jordan; 2Department of Applied Biological Sciences, Jordan University of Science and Technology, Irbid 22110, Jordan; 3Department of Public Health, Faculty of Medicine, Jordan University of Science and Technology, Irbid 22110, Jordan; 4Department of Ophthalmology, Faculty of Medicine, Jordan University of Science and Technology, Irbid 22110, JordanCorrespondence: Rami AlkhatibDepartment of Biotechnology and Genetic Engineering, Jordan University of Science and Technology, Irbid 22110, JordanTel +9622776319493Email rqalkhatib@just.edu.jo Background: Glaucoma is a neurodegenerative disease that leads to progressive loss of retinal ganglion cells, causing irreversible visual field defects. At the present time, glaucoma is clinically defined but the exact etiology is unknown. The aim of this study is to genotype rs2472493 and rs2487032 SNIPs within ABCA1 gene in 52 Jordanian Arab patients with primary glaucoma and 96 control subjects, and also to investigate the genetic association of these SNPs with primary glaucoma. Methods: DNA was extracted from both patients and controls according to a well-established procedure. Then, DNA was amplified by PCR using specific primers for this gene. Analysis of polymorphisms was carried out by using DNA sequencing genotyping method. Results: The results showed that the two SNPs (rs2472493 and rs2487032) located upstream of ABCA1 gene have no significant associations with primary glaucoma disorder (P > 0.05). Conclusion: This study is the first of its kind to reveal no genetic association between ABCA1 gene and primary glaucoma disorder in Jordanian population of Arab descent. [ABSTRACT FROM AUTHOR]

Various honey bees, especially subspecies Apis mellifera, occur in Africa and are distribute across the continent. The genetic relationships and identical genetic characteristics between honey bee subspecies in Africa (African bee subspecies) have not been widely investigated using sequence analysis. On the other hand, bioinformatics are developed rapidly and have diverse applications. It is anticipated that bioinformatics can show the genetic relationships and similarities among subspecies. These points have high importance, especially subspecies with identical genetic characteristics can be infected with the same group of pathogens, which have implications on honey bee health. In this study, the mitochondrial DNA sequences of four African subspecies and Africanized bees were subjected to the analyses of base composition, phylogeny, shared gene clusters, enzymatic digestion, and open reading frames. High identical base composition was detected in the studied subspecies, and high identical results from all tests were found between A. m. scutellata and A. m. capensis followed by A. m. intermissa and A. m. monticola. The least genetic relationships were found between A. m. lamarckii and the other subspecies. This study presents insights into the genetic aspects of African bee subspecies and highlights similarity and dissimilarity aspects. Also, Africanized honey bees derived from A. m. scutellata showed high genetic similarities to other African bees, especially A. m. capensis. Additionally, specific primers to identify these subspecies were designed and tested. [ABSTRACT FROM AUTHOR]

The identification of the ubiquitous spore‐forming bacterium Brevibacillus laterosporus, whose interest in pharma, agriculture, and other industrial sectors is raising, mostly relies on 16S ribosomal RNA gene sequence analysis. However, due to bacterial gene homology, this method appears insufficient for a proper discrimination of this species, so that the availability of other target genes is necessary. Leveraging the morphological and genetic feature uniqueness of B. laterosporus, a sensitive and reliable detection and quantification method based on polymerase chain reaction (PCR) and quantitative PCR assays, respectively, was developed. Targeting a highly conserved spore surface protein‐related gene, B. laterosporus could be easily found in different matrices including soil, food, and insect body. Primer set selectivity was confirmed to be very specific and no false positives or negatives were observed using DNA of different bacterial species as a template. The method developed is also suitable for the rapid identification of newly isolated B. laterosporus strains. [ABSTRACT FROM AUTHOR]

This article investigates some tensions in Brazil that cross the decade of 1980, known as the Constituent Moment (1985-1988). It analyses the trajectory and acting of the pro-Popular Participation Movement in the constitution (PPPM), without exhausting the subject, from different booklets of the period. Exercises as a theoreticalmethodological tool a look on affections, expectations and fears expressed in the sources. To this end, it uses the discussions by Cultural History of Politics (Thomas Mergel) and studies of passions in history (Pierre Ansart), therefore pointing the discursive conflict and political disputes in a time known in the country as "transition" that represented the passage between the Military Dictatorship to a democratization regime. [ABSTRACT FROM AUTHOR]

Abstract Steel panels must be protected during storage and transportation because they can corrode. This protection used to be afforded by chromate primers, but nowadays tannin-based primers are being studied. These primers are not only used as protection but also as adhesion promoter if the steel surface was not well prepared (presence of oxides or low roughness). The objective of this work was to prepare lanthanum "tannate", employing tara and quebracho tannins. The "tannates" were incorporated in primers formulation and applied on steel SAE 1010 panels (previously exposed to the humidity chamber or with 6 μm of roughness depth). These primed panels were exposed to humidity chamber where corrosion and blistering degrees were evaluated. Adhesion test were done. Electrochemical assays such as ionic resistance and corrosion potential measurements were carried out. Results showed that the adhesion and the barrier properties of the primers were good. Besides, on low roughness steel, the primers diminished the corrosion current. Results were compared with traditional zinc tetroxychromate primer. [ABSTRACT FROM AUTHOR]

Citrus bacterial canker (CBC) occurs due to Xanthomonas axonopodis pv. Citri (Xac) is a pathogenic bacteria. It was first reported in Japan, USA and many areas of India in 19th century. Synchronized by the spread of the causative agent the diversity of labelled pathogen continues to spread with new strains appearing in many other regions of the world. However, its diagnosis is quite difficult due to its symptoms similarity with other bacterial strains. This piece of study was designed to identify Xanthomonas axonopodis using polymerase chain reaction (PCR). PCR is more accurate, precise and fast method which is used for detection and quantification of many bacterial species. Two primers Xac01 and Xac02 were used for PCR based CBC detection. These primers were based on rpf gene region which give very specific results in Xac identification and can separate two much similar bacterial strains. There is another Primer pair designed on the pthA gene which act as universal primer and usually used for detection of all varieties of citrus canker bacteria. Leaves sample from different infected Citrus paradisi plants were collected for further analysis. Results confirmed that three samples among five have CBC. It is concluded that using PCR we can detect citrus canker disease more specifically which will help in discovering new eradication methods for its control. [ABSTRACT FROM AUTHOR]

Highlights • The taxon-specific primers were designed and evaluated in vitro and in silico for the phage identification. • The DNA markers identified in the present work can be useful for the taxon establishment of newly isolated phages. Abstract The phage taxonomy is primarily based on the morphology derived from Transmission Electron Microscopic (TEM) studies. TEM based characterization is authentic and accepted by scientific community. However, TEM based identification is expensive and time consuming. After the phage isolation, before analysis TEM, a DNA based rapid method could be introduced. The DNA based method could dramatically reduce the number of samples analyzed by TEM and thereby increase the speed and reduce the cost of identification. In the present work, four environmental phage isolates were identified based on TEM studies and genome size. The identification of these four phages was validated using DNA based method. The taxon-specific DNA markers were identified through multiple sequence alignments. The primers were designed at conserved genes (DNA polymerase or integrase) of 4 different phage taxa viz. family Ackermannviridae , genus Jerseyvirus , genus T4virus , and genus P22virus. These primers were evaluated using both in vitro and in silico approach for the amplification of the target taxons. Majority of the primer sets were found to amplify member species of the targeted taxa in vitro. In In silico analysis, six primer sets intended for identification of family Ackermannviridae showed positive amplification of ≥86.7% classified species. Further, the primers targeting the genus Jerseyviru s and T4viru s showed the amplification of 53.8% and ≥84.6% species, respectively. The present work is a case study performed to explore the possibility of use of taxon-specific primers for identification and taxonomic studies of newly isolated phages to supplement the TEM. [ABSTRACT FROM AUTHOR]

Several herbicidal chemicals are available for use in agriculture to increase the yield. Despite their usage commonly, they have impacts on crops. Related with this, in the current study, the effects of a widely used herbicide Roundup, which has glyphosate as the active ingredient. Molecular and spectroscopic approaches were combined together in order to investigate the effects of glyphosate on Zea mays. Depending on the EC50 value that reduce the root growth to 50%, three different concentrations of Roundup were used as 4 ml/L, 8 ml/L and 16 ml/L. They were applied to Z. mays seeds for 48 and 72 hours. For RAPD (Random Amplified Polymorphic DNA) PCR analysis; 8 of 10 RAPD primers produced band patterns and it was found that five RAPD primers among them produced polymorphic band patterns and then were used to produce a total of 61 bands. Value of polymorphisms was 32.78%. Genomic template stability changed in RAPD profiles at various Roundup concentrations. In addition to PCR analysis, the root samples were studied by Fourier transform infrared (FT-IR) spectroscopy in order to obtain the effects of Roundup on the quantity of biomolecules, membrane fluidity, cell wall components, lipid and protein structure of Z. mays. Each concentration of Roundup caused significant effects on Z. mays cells, particularly, the most effective concentration level was found to be 16 ml/L. Our corresponding results revealed that Roundup has prominent impacts on structure and function of biomolecules of Z. mays, all of which might be important for its growth. In conclusion, the combination of RAPD and spectroscopic analysis would offer efficient tool to determine the effects of any pesticides on plant cells. [ABSTRACT FROM AUTHOR]

What changes did the content, structure, and production of Russian primers published in the Soviet Union undergo between 1941 and 1948--that is, during the Second World War and its aftermath? This article answers this question by analyzing language, content, iconography, and the printing process. The first section addresses key characteristics of primers printed between 1941 and 1944, while the second section focuses on the content of postwar primers printed between 1945 and 1948. The final section addresses challenges facing the textbook approval and circulation process experienced by the State Pedagogical Publishing House of the Russian Soviet Federal Socialist Republic (RSFSR) from 1945 to 1948. [ABSTRACT FROM AUTHOR]

This introduction addresses the origins, general assumptions and intentions of the special issue. The guest editors show how reading primers published and used around the end of the Second World War in several European countries may serve as an object of study in different disciplinary contexts. They present a broad working definition of the reading primer as an educational medium that lends itself to interdisciplinary research which takes into account aspects such as visual and textual content, materiality, and societal contexts of production, distribution and usage. The editors further highlight characteristics of current research into primers and argue in favor of more comparative approaches that reveal transnational dimensions of textbooks designed to teach children how to read and write. [ABSTRACT FROM AUTHOR]