Primer

Objective Plants that have a symbiotic relationship with arbuscular mycorrhizal fungi (AMF) compared with non-mycorrhizal plants have a higher ability to resist against various biotic and abiotic environmental stresses such as drought, salinity, pests, and plant diseases. This study aim is studying different species of AMFs in Kerman province. The identification of these microorganisms is to provide the basis for further studies on the use of these microorganisms in germination, growth, and proliferation of medicinal plants. Methods To molecular and morphological identification of arbuscular mycorrhizal fungi, sampling was done from the rhizosphere of medicinal plants in some areas of Kerman province. Arbuscular mycorrhizal fungi spores were isolated from the sampled soils by the wet
sieve method. The grouping was done based on spore microscopic slides and
morphological characteristics such as spore color, shape, surface ornamentation, size,
wall structure and spore characteristics in water. DNA was extracted from single spores.
A Part of ribosomal DNA was amplified by nested PCR using SSUmaf and LSUmAr
primers in the first step, SSUmCf, and LSUmBr in the second step. After horizontal
agarose gel electrophoresis and see the related band the PCR product was sent for
sequencing. Then sequencing results were analyzed using BLAST, Gene Runner and
Clustal Omega Softwares, and the phylogenetic tree was plotted using MEGA 7 software.
Results
According to morphological and molecular studies, Viscospora viscosa, Septoglomus
deserticola, Funneliformis caledonium, Funneliformis mosseae, and Diversispora
spurca were identified. Finally, the existence and manner of species identity were
investigated in the phylogeny tree.
Conclusion
Since the morphological features used in the identification and classification of
mycorrhizal fungi are limited, molecular methods can be used for safer classification, and
in this way, Recognize differences of Similar species morphologically and similarity of
different species in appearance.

A conventional PCR and a SYBR Green real-time PCR assays for the detection and quantification of Phytophthora cryptogea, an economically important pathogen, have been developed and tested. A conventional primer set (Cryp1 and Cryp2) was designed from the Ypt1 gene of P. cryptogea. A 369 bp product was amplified on DNA from 17 isolates of P. cryptogea. No product was amplified on DNA from 34 other Phytophthora spp., water moulds, true fungi and bacteria. In addition, Cryp1/Cryp2 primers were successfully adapted to real-time PCR. The conventional PCR and real-time PCR assays were compared. The PCR was able to detect the pathogen on naturally infected gerbera plants and on symptomatic artificially infected plants collected 21 days after pathogen inoculation. The detection limit was 5 × 103 P. cryptogea zoospores and 16 fg of DNA. Real-time PCR showed a detection limit 100 times lower (50 zoospores, 160 ag of DNA) and the possibility of detecting the pathogen in symptomless artificially infected plants and in the re-circulating nutrient solution of closed soilless cultivation systems.

Создание латгальских Букварей прошло сложный путь от религиозных книг, основанных на Катехизисе и популярных молитвах, до современного Букваря. Менялись их названия, происходила систематизация самого латгальского языка. Это заметно даже на примере названий одного и того же учебника при его переизданиях. Например, «Eysa mociba del masu bernu» (1835), «Eysa mociba del mozu bernu» (1839), «Eysa mociba del mozu barnu» (1849) и т.д.
Роль белорусского фактора в истории создания и распространения латгальского учебного книгоиздания нам кажется весьма существенной. Некоторые из авторов первых латгальских учебников были уроженцами Беларуси (например, Т. Кучинский), получили образование и трудились в Беларуси до переезда в Латгалию (например, М. Рот). Первые латгальские Буквари были изданы в столице белорусско-литовского государства – Вильне, а после также в белорусских типографиях в Могилеве и Полоцке. Лишь со второй половины 19 в. Буквари на латгальском письменном языке начинают издаваться собственно в Латгалии. На протяжении всего 19 – начала 20 в. развитие учебной книжности в Латгалии происходило под контролем администрации Витебской губернии и Виленского генерал-губернаторства. Крайне важную роль в создании первых латгальских Букварей сыграли преподаватели и выпускники Виленской и Полоцкой академий, Краславской семинарии и других учреждений, самым тесным образом связанных с Беларусью.
Как и белорусские издания, латгальские Буквари имели малый (карманный) формат и, обычно, небольшой объем. В отличие от латышских изданий из Курляндии и Лифляндии, они издавались не готикой, а распространенной в ВКЛ антиквой. Следует отметить и еще одну общую черту: первые Буквари на латгальском и белорусском языках издавались латинским шрифтом на основе правил польской латиницы и лишь в начале 20 в. оба народа окончательно перешли на национальные нормы латиницы, в которых для передачи специфических звуков использовались знаки с диакритикой (например, «SZ» перешло в «Š», «CZ» – в «Č» и т.д.).
Все латгальские Буквари того времени были построены на основании так называемого буквослагательного метода, распространенного в учебной практике Беларуси, Латвии, Литвы, Польши и других стран. Конечно же, белорусские и латгальские Буквари роднило содержание: их основу составляли религиозные тексты (Катехизис, молитвы, поучения и пр.). В этом отношении они были похожи друг на друга значительно больше, чем на Буквари иных народов.
Первые учебники создавались католическими священниками, преимущественно монахами-иезуитами из Вильны и Полоцка. Вероятно, авторы-иезуиты благополучно усвоив практику педагогической работы и учебного книгоиздания в ВКЛ, активно внедряли ее в отношении населения Латгалии. В дальнейшем практику работы иезуитов переняли базилиане, миссионеры и пиары из Витебской и Виленской губерний.
После восстания 1830–1831 гг. царская власть начала ограничивать влияние католической церкви на просвещение населения как в Беларуси, так и в Латгалии, а после восстания 1863–1864 гг. и вовсе был взят курс на массовую русификацию населения «Северо-Западного края» Российской империи. Для белорусов, латгальцев, латышей, литовцев и иных жителей края настойчиво предлагалось обучение исключительно на русском языке. И лишь в начале 20 в., когда был снят запрет на развитие национальной письменности, белорусские и латгальские Буквари вышли из подполья и начали издаваться официально. С этого времени их пути идут самостоятельно, хотя во многом параллельно.
И все же понимание общности истории наших культур имеет огромное значение именно сегодня, ведь оно не только свидетельствует о имеющемся опыте конструктивного и плодотворного взаимодействия на протяжении многих веков, но и является почвой для развития сотрудничества в настоящее время.

Six primers for the amplification of three non-coding regions of chloroplast DNA via the polymerase chain reaction (PCR) have been designed. In order to find out whether these primers were universal, we used them in an attempt to amplify DNA from various plant species. The primers worked for most species tested including algae, bryophytes, pteridophytes, gymnosperms and angiosperms. The fact that they amplify chloroplast DNA non-coding regions over a wide taxonomic range means that these primers may be used to study the population biology (in supplying markers) and evolution (inter- and probably intraspecific phylogenies) of plants.

The object of this analysis was to compare the organic and the conventional farming on the basis of primer database and modeling at the location of Hortobágy that is a well-known organic farming region in Hungary. 20 model variations were worked out for the conventional and 20 for the organic farming situation. According to the results, there is no significant difference considering the costs per hectare in the two modeled farming methods, although the structures of the production costs show huge deviations. The cost per production unit is typically higher in the organic model. Considering most of the model variations, organic farming is proved to be more profitable, but this economic pre-eminence is not explicit by any means.

Introduction: Inpatient training and evidence-based medicine (EBM) curricula are fundamental components of medical education. Teaching EBM And Clinical topics in the Hospital (TEACH) Cards is an inpatient curricular tool developed to help guide efficient, discussion-based teaching sessions. TEACH Cards aims to increase frequency of inpatient teaching, improve exposure to the breadth of inpatient topics, advance EBM skills, and improve efficiency in answering clinical questions. Methods: TEACH Cards is a set of 25 topic-based cards, each addressing an adult inpatient medicine topic by asking background questions and encouraging learners to write and answer foreground questions. Residents and faculty from a family medicine residency rotating on an adult inpatient medicine service during the 6-month study period were invited to complete a prerotation survey, use the TEACH Cards, and then complete a postrotation survey. Results: Out of 54 potential participants, 35% completed both the p...

The teak Catalase gene was amplified and cloned using consensus PCR primers designed based on sequences from other plant species. The obtained teak Cat DNA sequences were used to develop specific primers. The specific primer set could successfully amplify a specific DNA fragment from teak in all populations studied. Using PCR-SSCP, 5 different alleles were detected. The tested nuclear gene primer set had considerable potential as a DNA marker for population analysis in teak. This approach could be used to specifically amplify fragments in other plant species and applied to study evolution, population genetics, outcrossing rate and mating patterns.

In the present work we evaluateTrypanosoma cruziDNA detection by PCR using the nuclear oligonucleotides BP1/BP2 as primers. These primers are targeted to the 5′ and 3′ ends of the coding region for the flagellar protein F29. An amplification product of BP1/BP2 is a DNA band 692 bp long. Titration assays were performed to evaluate the minimum amount of parasite DNA that can be detected by this assay, resulting in 10 fg (equivalent to about 1/20 of the genome). The assay was also performed usingT. cruziDNA from different strains, clones, and human-derived isolates obtaining, in all cases, amplification products. No DNA amplification was observed when the PCR was performed using DNA fromLeishmania braziliensis,but whenT. rangeliDNA was used, a 615-bp-long fragment was amplified. Under appropriate gel conditionsT. cruziandT. rangeliDNA amplicons could be differentiated. When both conventional xenodiagnosis and PCR detection of parasite DNA in the feces of insect vectors fed with blood from infected patients were compared, 10 of 20 samples were positive by both techniques. However, 2 other samples with positive serology were also positive by PCR. When PCR was performed on blood samples from infected and uninfected individuals, 62 of 65 serologically positive human samples amplified the BP1/BP2 692-bpT. cruziDNA fragment (sensitivity >95%). The 3 negative samples were positive when Southern blot hybridization was performed using the radiolabeled PCR amplification product as probe (sensitivity 100%).

Introduction: Family medicine residency programs (FMRPs) endeavor to meet evidence-based medicine (EBM) subcompetencies through the milestones project. Comprehensive descriptions of clinical pharmacists’ contributions in teaching EBM within the context of residency are limited.  Methods: Over a study period of 2 months, clinical pharmacists across five FMRPs in four states were invited to track their interactions with physician residents. EBM resources, skills, and targeted milestone data were collected. Pharmacists also quantified their nonpatient care contributions to EBM.  Results: Of the 16 clinical pharmacists invited, 16 (100%) participated in the October and 12 (75.0%) in the March collection period. A total of 598.9 half days over 2 months (42 working days) of available teaching time were reported. The tracking tool captured 1,253 EBM teaching encounters with a total average of 2.1 encounters per half day. Of those encounters, point-of-care references were most commonly used...

Introduction Care of patients with chronic medical and mental health conditions can be a source of frustration for primary care clinicians and may present a challenge in modeling effective interventions for medical learners. Mind-body medicine (MBM) interventions have shown success for a variety of conditions, and training in MBM has been associated with decreased burnout and improved professional satisfaction.8 We piloted MBM collaborative visits led by faculty physicians and facilitated by medical learners. We then assessed their efficacy treating patients with complex needs. Methods We conducted a series of eight weekly 2.5-hour MBM interventions for groups of five to eight participants (52 in total) with chronic health conditions. Matched-pair hypothesis t tests analyzed nine health indicators measured pre- and postintervention: the Patient Health Questionnaire-9 (PHQ-9) as well as participants' perceived mental and physical health, stress and stress coping, agency, and capa...

In the present study, the effect of primer and slurry composition on the washcoat characteristics of γ-alumina on stainless steel substrate has been investigated. Washcoating was done following a two-step procedure: primer coating followed by slurry coating. Washcoat was characterized by SEM, adherence test and BET surface area measurement. For coating of the primer, Disperal with or without polyvinyl alcohol (PVA) was used. Without PVA, the sol settled in the microchannels whereas at high concentrations of PVA, the sol was too viscous. The optimum primer composition was found to be 2% Disperal and 4% PVA. In the subsequent slurry coating step, various properties of the slurry such as pH, viscosity, solid loading were optimized to obtain well-adhered, uniform washcoat on stainless steel substrate. PVA was added to the slurry to increase the binding and also enhance the viscosity, resulting in better adherence. An adverse effect of adding PVA was a reduction in the surface area. To r...

Many different single nucleotide polymorphisms (SNPs) genotyping methods have been developed recently. However, most of them are expensive. Using restriction enzymes for SNP genotyping is a cost-effective method. However, restriction enzyme mining for SNPs in a genome sequence is still challenging for researchers who do not have a background in genomics and bioinformatics. In this review, the basic bioinformatics tools used for restriction enzyme mining for SNP genotyping are summarized and described. The objectives of this paper include: i) the introduction of SNPs, genotyping and PCR-restriction fragment length polymorphism (RFLP); ii) a review of components for genotyping software, including tools for primer design only or restriction enzyme mining only; iii) a review of software providing the flanking sequence for primer design; iv) recent advances in PCR-RFLP tools and natural and mutagenic PCR-RFLP; v) highlighting the strategy for restriction enzyme mining for SNP genotyping;...

Truffles are hypogeous ectomycorrhizal fungi. They belong to the genus Tuber and are currently considered a hot spot in fungal biology due to their ecological and economic relevance. Among all the species, Tuber magnatum is the most appreciated because of its special taste and aroma. The aim of this work was to set up a protocol to detect T. magnatum in soil and to assess its distribution in a natural truffle-ground. We used the β-tubulin gene as a marker to identify T. magnatum in the soil. This gene allowed us to trace the distribution of the fungus over the entire truffle-ground. Tuber magnatum was found, in one case, 100 m from the productive host plant. This study highlights that T. magnatum mycelium is more widespread than can be inferred from the distribution of truffles and ectomycorrhizas. Interestingly, a new haplotype – never described from fruiting body material – was identified. The specific detection of T. magnatum in the soil will allow to unravel the ecology of this fungus, following its mycelial network. Moreover, this new tool may have practical importance in projects aimed to increase large-scale truffle production, checking for T. magnatum persistence in plantations.