Comparison of Time Course Detection of Human Male DNA from Blood Stains on Various Objects on Surface in a Natural Environment and in a Laboratory Using Loop-Mediated Isothermal Amplification (LAMP)

In forensic study, the biological evidence can easily degrade, especially DNA. Degraded and environmentally challenged samples can produce numerous problems in forensic DNA analysis including loss of band product. Loop-mediated isothermal amplification or LAMP is one of the DNA analysis techniques used in forensic study. This study explores the limitations of the efficiency of the LAMP technique on abandoned DNA. For the DNA template, 8 male and 2 female blood-stained samples were taken from the surfaces, namely, brick, cloth, and tile from inside, and buried outside the laboratory. The LAMP reaction was used to amplify the SRY gene for detecting male DNA. All the blood-stained samples were stored for 1, 7, 15, 30, and 45 day (s). The LAMP product from the blood-stained samples on all the surfaces that were kept in a laboratory was detected using the gel electrophoresis technique from day 1 until day 45. However, the LAMP product on day 30 and 45 was smear and dim. The LAMP product from the blood-stained samples buried outside the laboratory was observed using the gel electrophoresis technique within day 30 (smear and dim). To increase the efficiency of detection, the qLAMP technique detected product on all the male samples from all the surfaces buried outside the laboratory for 45 days. The results indicate that this LAMP condition was possible detecting male DNA and the environmental factors are the main influence on the sensitivity of the LAMP technique. In addition, the qLAMP technique can increase the capacity and sensitivity of the detection.

EXTRACTION AND PURITY DNA OF Culex spp MOSQUITO IN KEMELAK VILLAGE, BINDUNG LANGIT, OGAN KOMERING ULU

Culex spp are mosquito vectors that have a very wide distribution capability and are carriers of pathogens that can interfere with human and animal health. The wide distribution makes Culex spp a dangerous threat. DNA extraction is one of the important steps in obtaining genetic information and genetic analysis. Good quality DNA is used for activities such as the use of molecular markers, genome library creation, and sequencing. This study aims to determine the quality, concentration and purity of Culex spp mosquito DNA in Kemelak Bindung Langit Village, OKU Regency. It is hoped that the sample can be used for further research analysis on Mitochondria D-Loop Sequences in Culex spp mosquitoes. Quantitative measurement of DNA in the form of concentration and purity of DNA using Nanodrop Thermo cycle while qualitative DNA using electrophoresis technique. The results of the isolation of the mosquito genome DNA, obtained clear DNA bands without any degradation (smear) and the concentration results for the four samples ranged from 10-100 ng/µL and the DNA purity was good, ranging from 1.8 to 2.00.

Sample stacking – Capillary electrophoretic determination of nitrate and nitrite contents as nitric oxide metabolites in honey varieties originated from Anatolia

Nitrate and nitrite ions taken from food are the sources of bioavailable nitric oxide (NO) in the nitrogen cycle. Some beneficial effects of honey on health are attributed to the ability of honey to increase NO production. The variation of nitrate and nitrite levels of honey samples collected from different Anatolia regions were clarified using capillary electrophoresis technique. The sensitivities of both anions were improved with the application of the sample stacking method. Separation buffer consisted of 30 mmol L−1 formic acid and 30 mmol L−1 sodium sulfate at a pH of 4.0. The CE technique revealed that 18 honey samples contained nitrate anion ranged between 2.53 and 31.8 mg kg−1. Nitrite amounts were found in lower amounts in the honey samples as between non-detected and 0.533 mg kg−1. The observed differences in nitrate levels between honey varieties may be a way to determine honey's origin.

Cytosolic 5′-Nucleotidase II Silencing in Lung Tumor Cells Regulates Metabolism through Activation of the p53/AMPK Signaling Pathway

Cytosolic 5′-nucleotidase II (cN-II) is an allosteric catabolic enzyme that hydrolyzes IMP, GMP, and AMP. The enzyme can assume at least two different structures, being the more active conformation stabilized by ATP and the less active by inorganic phosphate. Therefore, the variation in ATP concentration can control both structure and activity of cN-II. In this paper, using a capillary electrophoresis technique, we demonstrated that a partial silencing of cN-II in a pulmonary carcinoma cell line (NCI-H292) is accompanied by a decrease in adenylate pool, without affecting the energy charge. We also found that cN-II silencing decreased proliferation and increased oxidative metabolism, as indicated by the decreased production of lactate. These effects, as demonstrated by Western blotting, appear to be mediated by both p53 and AMP-activated protein kinase, as most of them are prevented by pifithrin-α, a known p53 inhibitor. These results are in line with our previous observations of a shift towards a more oxidative and less proliferative phenotype of tumoral cells with a low expression of cN-II, thus supporting the search for specific inhibitors of this enzyme as a therapeutic tool for the treatment of tumors.

Occurrence of Meloidogyne arenaria in black pepper (Piper nigrum L.) in the extreme south of the State of Bahia, Brazil

Summary Samples of black pepper root with the presence of galls from the most southern region of the State of Bahia, Brazil, were characterized biochemically and morphologically using three criteria: i) observation of the anterior region of the males; ii) analysis of female perineal configuration and iii) polyacrylamide gel electrophoresis technique. Meloidogyne arenaria was found. This is the first report of this pathosystem in the State.

Protein corona profile of graphene oxide allows detection of glioblastoma multiforme using a simple one-dimensional gel electrophoresis technique: a proof-of-concept study

Protein Corona-based blood test for the detection of glioblastoma multiforme.

openPFGE: An open source and low cost pulsed-field gel electrophoresis equipment

DNA electrophoresis is a fundamental technique in molecular biology that allows the separation of DNA molecules up to ~50 Kbp. Pulsed-field gel electrophoresis [PFGE] is a variation of the conventional DNA electrophoresis technique that allows the separation of very large DNA molecules up to ~10 Mbp. PFGE equipment is very expensive and it becomes an access barrier to many laboratories. Also, just a few privative designs of the equipment are available and it becomes difficult for the community to improve or customize their functioning. Here, we provide an open source PFGE equipment capable of the separation of DNA molecules up to, at least, ~2 Mbp and at low cost: USD$850, about 3% of the price of typical commercial equipment.Specifications table

HAPTOGLOBIN POLYMORPHISM AND CHRONIC TENDINOPATHY

Introduction: Tendinopathy is a pathology of tendons of multifactorial origin displayed clinically as chronic degenerative tissue resulting in dysfunction and painful musculoskeletal system. Genetic implications in tendinopathy are not fully elucidated. Haptoglobin (Hp) polymorphism has been investigated for potential role in several diseases such as cardiovascular diseases. This study aims at investigating Hp polymorphism potential association with chronic tendinopathy in Jordan. Materials and Methods: A hospital-based case-control study, of 101 patients diagnosed with chronic tendinopathy and 104 age and gender matched controls, was employed and Hpphenotypes distributions were determined using Sodium Dodecyl Sulphate Polyacrylamide gel electrophoresis technique. Results: showed a significant association between tendinopathy cases and controls in relation to Hpphenotypes frequencies (Fisher’s Exact Test, [Formula: see text]). Odd ratios showed that the odds of tendinopathy cases in Hp2.1 group were reduced while odds in Hp2.2 and Hp1.1 were increased by 3.22 and 7.34 times, respectively, as likely compared to the controls. Conclusion: Results suggested that Hppolymorphism might play a role in chronic tendinopathy genesis.