The appearance of DNA bands pattern based on the result of primary selection of RAPD orchid Phaius spp.

This study aimed to examine the appearance of the DNA band pattern resulted from the selection of RAPD primers on Phaius spp. namely Phaius tankervillae, Phaius montanus, Phaius collasus and Phaius amboinensis. The research material was performed in the Center for Plant Conservation of the LIPI Bogor Botanical Gardens. Molecular analysis was carried out at the Laboratory of the Center for Horticultural and Tropical Studies IPB using RAPD. The study showed that the 2 primers RAPD OPA 02 and OPA 16 can be used for DNA amplification of orchids Phaius spp (Phaius tankervillae, Phaius montanus Phaius collasus and Phaius amboinensis) because they produce clear DNA bands. The result of PCR amplification on Phaius tankervillae, Phaius montanus, Phaius collasus, and Phaius amboinensis using OPA 02 and OPA 16 primers produced 11 and 9 DNA bands, respectively, with an average of 5 DNA bands per primer. In the band pattern at 800 bp on OPA 02 primers resulting sharp and clear band pattern quality.

Vnir reflectance spectroscopyof Mercury like glasses

<p>Glassy materials have been recognized over Mars, Moon and many different meteorites (Farrand et al. 2016; Delano 1986; Varela & Kurat 2004). Planetary glasses result from impact events but they are also found as volcanic products (Farrand et al 2016). Morlock et al. (2017) and Morlok et al. (2021) investigated by means of different experimental techniques (bi-directional diffuse reflectance FTIR, in situ FTIR microscopy, Raman, EPMA and optical microscopy) a suite of synthetic samples with composition similar to those inferred for different Hermean terrains. Here we extended the study of the same materials to the VNIR region (bidirectional reflectance spectroscopy: 350 to 2500 nm). We analyzed 8 different samples with different chemical compositions, produced under different oxygen fugacity conditions We prepared eight granulometric classes between 0 and 250 μm, namely: 0-25; 25-63; 63-100; 100-125; 125-150; 150-180; 180-200 and 200-250 μm. The dominant feature in the VNIR region is due to the Fe absorption band at about 1 μm accompanied, in the more oxidized samples, by a smaller feature at 480 nm likely due to ferric oxide development. Iron free samples (FeO < 0.1 wt%) show characteristic spectral shapes with a distinctive feature at about 640 nm attributable to TiO2. Even for very low FeO content, it is possible to observe a weak yet clear band at about 900-1000 nm due to Fe absorption which explain the dominance of the spectral features due to Fe absorption at higher FeO content. Additional small bands at higher wavelengths (1300-1400 and 1900 nm) suggest a low content of water and/or –OH species in the samples. We investigated the spectral features as a function of composition, grain size and oxidation in order to gain as much information as possible on the nature of the spectra and compare them with remote sensing data or meteorites VNIR comparison. Our data on synthetic and realistic Hermean compositions will allow a better understanding of remotely acquired VisNIR spectra, which will be particularly helpful in view of the upcoming beginning of the BepiColombo ESA/JAXA mission.</p> <p> </p> <p>Acknowledgments: The authors acknowledge financial contribution from the Italian Space Agency (ASI) under ASI-INAF agreement 2017-47-H.0 (Simbio-SYS). CC, EB are also supported by agreement ASI-INAF n.2018-16-HH.0.</p>

Expression of Proteinase-Activated Receptor 2 During Colon Volvulus in the Horse

Large colon volvulus in horses is associated with a poor prognosis, especially when ischemic-reperfusion injury of the affected intestinal tract develops. Proteinase-activated receptor 2 (PAR2) plays an important role in the pathogenesis of inflammation in the gastrointestinal tract. The aim of this study was to evaluate the distribution and expression of PAR2 in colonic pelvic flexure of horses spontaneously affected by large colon volvulus (CVH group). Eight horses admitted for severe abdominal colon volvolus and which underwent surgery were included. Colon samples were collected after enterotomy. Data previously obtained from healthy horses were used as a control group. Histologic evaluation was carried out to grade the severity of the colon lesions. Immunofluorescence, western blot and quantitative polymerase chain reaction (RT-qPCR) were carried out on colon samples to evaluate PAR2 expression. In addition, the transcriptional profile of cytokines and chemokines was evaluated using RT2 Profiler™ PCR Array Horse Cytokines & Chemokines. Three out of the eight patients were euthanised due to clinical deterioration. Immunostaining for PAR2 was observed in the enterocytes, intestinal glands and neurons of the submucosal and myenteric plexi. In the CVH horses, the expression of PAR2 mesenger RNA (mRNA) did not differ significantly from that of the healthy animals; western blots of the mucosa of the colon tracts showed a clear band of the expected molecular weight for PAR2 (~44 kDa) and a band smaller than the expected molecular weight for PAR2 (25kDa), suggesting its activation. The gene expressions for C-X-C motif ligand 1 (CXCL1); interleukin 8 (IL8), macrophage inflammatory protein 2 beta (MIP-2BETA) were upregulated in the colic horses as compared with the colons of the healthy horses. Therefore, in the present study, the expression and activation of PAR2 in the colons of horses in the presence of an inflammatory reaction like that occurring in those with spontaneous colon volvulus was confirmed.

Eliminating the isolated states from the band structure of doped TiO2: A first principle study

Band structure of anatase [Formula: see text] could be modified by doping foreign atoms. With ab-initio calculations, considering praseodymium, nitrogen and gadolinium as potential dopants, some mono-, co- and tri-doped models are introduced. The structure modification due to each model is compared with a standard [Formula: see text] system. Pr doping reduced the band gap of [Formula: see text] by introducing Pr 4f states below the conduction band, and the Fermi level appears below the conduction band minimum. The addition of nitrogen in Pr-doped [Formula: see text] pushed back the Fermi level to its intrinsic position while reducing the band gap. The tri-doped system comprising of Pr, N and Gd as dopants provided a populated band gap. The impurity states associated with Gd appeared in the middle of the band gap. The density of states analysis reveals that Pr 4f states change the location of N 2p states in the band structure. Instead of appearing as isolated states, the N 2p states are mixed with O 2p states while the Pr 4f states are coupled with the Ti 3d states in Pr, N-[Formula: see text]. The Pr, N, Gd tri-doped [Formula: see text] system provided strong visible light absorption due to the stepwise transition of electrons by Gd f states. While providing reasonable visible light absorption, the Pr, N-[Formula: see text] is expected to provide strong visible light photoactivity due to the clear band gap. Due to the absence of isolated states, the generated photo-excited carriers would be efficiently utilized in the oxidation/reduction processes.

On the electronic structure of methyl butyrate and methyl valerate

We present novel results of the analysis of the electronic structure of two aliphatic esters: methyl butyrate and methyl valerate. High-resolution photoabsorption spectra were collected and analyzed over the energy range 4.0–10.8 eV and showed for both the molecules not only a clear band of the HOMO to LUMO transition, but also vibronic structure associated with the first Rydberg-valence transition. Photoelectron spectra recorded from 9 to over 28 eV revealed many ionization states with the first adiabatic ionization energies found to be 9.977 eV and 9.959 eV for methyl butyrate and methyl valerate, respectively. Ab initio calculations have been performed in order to help assign the photoabsorption and photoelectron features. Photolysis life times in the atmosphere were calculated revealing that photolysis is not competitive over hydroxyl radical scavenging in the process of removal of these esters from the atmosphere. Graphical abstract

PCR and Sequencing Base Detection of Gummosis Disease on Citrus aurantifolia Caused by Lasiodiplodia theobromae and Evaluation of Its Antagonisms

Diplodia gummosis disease caused by Lasiodiplodia theobromae is an economically important postharvest fruit decay that occurs on all types of citrus grown in Bangladesh. The present investigation was conducted to isolate and identify the pathogenic fungus responsible for postharvest diplodia gummosis disease (DGD) of citrus as well as the evaluation of its biological control through microbial antagonists. DGD causing pathogen was identified by physiological, morphological and molecular methods. The pathogenic fungus was isolated from the surface of postharvest lime fruits. The optimum growth of the fungi was observed in maltose, 2% NaCl, 15% sugar and pH 7 containing PDA medium at 35⁰C. PCR products of the internal transcribed spacer (ITS) region of the fungus showed approximately 650 bp size clear band in gel electrophoresis. The amplified region of the fungus showed 99.62% similarities with the sequences of L. theobromae. Artificially inoculation of the fungus in malta, musumbi, sweet orange, lime, and guava fruits showed similar size clear band and typical diplodia gummosis disease symptom. Methanol extracts of Datura metal displayed the highest inhibition (75.25%) against the isolated fungus. Non-pathogenic fungi Trichoderma viride showed the highest antagonistic efficiency followed by Neofusicoccum mangifera against the isolated fungus. The tested soil bacteria did not show significant antagonistic activity against the isolated fungus. Therefore, the DGD of citrus control system should be integrated into the overall citrus postharvest decay control system to reduce all citrus postharvest diseases and to protect fresh citrus fruit values.

Recombinant cold shock domain containing protein is a potential antigen to detect specific antibody during early and late infections of Haemonchus contortus in goat

Background: Haemonchus contortus (H. contortus) is the most abundant nematode causing haemonchosis with major economic losses to the small ruminant industry farming worldwide. Effective prognosis and treatment depend upon the early diagnosis of H. contortus infection. To date, no widely-approved methods are available to identify prepatent H. contortus infection. The aim of this study was to evaluate the diagnostic potential of recombinant cold shock H. contortus protein (rHc-CS) during early and late infections of H .contortus in goat. Results: Purified rHc-CS exhibited a clear band, with a molecular weight about 38 kDa. No eggs of H. contortus were detected in feces collected at 14 days post infection. However, Specific anti rHc-CS antibodies were detectable in sera of all infected goats during early stage (2nd week of infection) and late stage (3rd to 14th week of infection) using immunoblotting assay. Furthermore, no cross reactivity was observed against most commonly found pathogens (Trichinella spiralis, Fasciola hepatica, and Toxoplasma gondii) and uninfected goats. The format variables for rHc-CS indirect-ELISA were optimized. The optimum antigen coating concentration was found 0.28μg/well at 37℃ 1h and overnight at 4°C. Optimum dilution ratio of serum and rabbit anti-goat IgG was recorded 1:100 and 1:4000 respectively. The best blocking buffer was 5% bovine serum albumin (BSA) and the best time for blocking, serum incubation and TMB reaction was recorded as 60, 120 and 10 minutes respectively. The cut-off value for positive and negative interpretation was determined as 0.352 (OD450). The diagnostic specificity and sensitivity of the rHc-CS, both were recorded 100%. Conclusion: These results demonstrated that rHc-CS is a potential immunodiagnostic antigen to detect specific antibodies at early and late H. contortus infections in goat.

ANALYSIS OF nrLSU GENE TO SUPPORT IDENTIFICATION OF FUNGUS BELONGING TO Cordyceps GENUS AND CLAVICIPITACEAE FAMILY

Nucleotide sequences of the nuclear large ribosomal subunit (nrLSU) have been used in fungal systematics for a long time. nrLSU was also used in Cordyceps and related genera within the Clavicipitaceae family. A previously identified sample by morphology and ITS was used in this research to analyze the ability of nrLSU to support the identification of entomopathogenic fungi. Our results show that we successfully amplified nrLSU gene using the primer pair LR0R and LR5. The PCR product on agarose gel showed a clear band at 950 bp. Sequencing method was then adopted and proofread before molecular phylogenetic analysis was applied with reference sequences obtained from the publication of Sung et al. Once again, this analysis confirms the DL0004 specimen as Cordyceps neovolkiana.

Optimasi Konsentrasi DNA dan MgCl2 pada Reaksi Polymerase Chain Reaction-Random Amplified Polymorphic DNA untuk Analisis Keragaman Genetik Tanaman Faloak (Sterculia quadrifida R.Br) (Optimization of DNA and MgCl2 Concentrations in Polymerase Chain Reacti

Abstrak Faloak merupakan tanaman yang tumbuh di lahan kritis. Sebagai upaya mendukung pemuliaan dan konservasi tanaman faloak diperlukan informasi keragaman genetiknya. Salah satu metode analisis keragaman genetik adalah menggunakan penanda DNA yang berbasis PCR. Untuk itu diperlukan kondisi PCR (Polymerase Chain Reaction) yang tepat sehingga diperoleh hasil yang dapat dianalisis lebih lanjut. Penelitian ini bertujuan menentukan kondisi optimum PCR-RAPD (Polymerase Chain Reaction-Random Amplified Polymorphic DNA) tanaman faloak. Ekstraksi DNA dilakukan dengan metode CTAB. Optimasi dilakukan dengan menggunakan beberapa konsentrasi DNA cetakan dan MgCl2. Kondisi optimum PCR-RAPD tanaman faloak yang menghasilkan pita produk PCR yang jelas diperoleh menggunakan 50 ng/ul DNA, 3 mM MgCl2 serta jumlah siklus termal 45 x. Kata kunci : PCR-RAPD, optimasi, tanaman faloak Abstract Faloak is a plant that grows on critical lands. In an effort to support breeding and conservation of faloak, information about its genetic diversity is required. One of the methods of genetic diversity analysis is using PCR-based DNA markers. For that purpose, proper PCR conditions is needed in order to obtain results that can be further analyzed. This study aimed to determine the optimum conditions for PCR-RAPD of faloak plants. DNA extraction was conducted using CTAB. Optimization was done by using several concentrations of DNA templates and MgCl2. The optimum conditions of PCR-RAPD of faloak plants that produce clear band of PCR products were obtained using 50 ng/ ul DNA, 3 mM MgCl2 and 45x thermal cycles Keywords : PCR-RAPD, optimization, faloak plant

Optimization of PCR Conditions for DNA Amplification of Common Buckwheat Using EST Primers

Under optimal conditions the PCR reaction is very efficient; microgram quantities may be synthesized from a single molecule of substrate DNA. DNA of four lines of common buckwheat (Kyusu, Canada, Miyazaki and Botansoba) was used to optimize PCR reaction and cycling program of 26 primers for DNA amplification of common buckwheat. Annealing temperature (Ta), PCR cycle number and MgCl2 concentration were considered optimum if the single clear band was observed. Of the 26 primers Ta of only 10 primers could be optimized. Three primer pairs performed best at Ta of 54°C. The optimum concentration of MgCl2 was found to be 1.5mM for all primer pairs. Similarly the number of PCR cycles was found to be 40 for all 10 primer pairs except for primer pair 57. Optimized PCR conditions were used for subsequent studies such as transferability of EST primers to other Fagopyrum species and construction of linkage map.Nepal Agric. Res. J. Vol. 8, 2007, pp. 1-6DOI: http://dx.doi.org/10.3126/narj.v8i0.11563