Accurate determination of DNA yield from individual mosquitoes for population genomic applications

Abstract  Accurate estimates of DNA quantity are likely to become increasingly important for successful genomic screening of insect populations via recently developed, highly multiplexed genotyping assays and high-throughput sequencing methods. Here we show that genomic DNA extractions from single Anopheles gambiae Giles using a standard commercial kit-based methodology yield extracts with concentrations below the linear range of spectrophotometric absorbance at 260 nm. Concentrations determined by spectrophotometry were not reproducible, and are therefore neither accurate nor reliable. However, DNA quantification using a fluorescent nucleic acid stain (PicoGreen^®) gave highly reproducible concentration estimates, and indicated that, on average, single mosquitoes yielded approximately 300 ng of DNA. Such a total yield is currently insufficient for many high-throughput genome screening applications, necessitating whole genome amplification of all or most individuals in a population prior to genotyping.     

Ultrastructural changes in the dermal chromatophore unit of Hyla arborea during color change

Summary The structural changes in the chromatophores of Hyla arborea related to changes in skin color were studied by electron microscopy and reflectance microspectrophotometry. During a change from a light to a darker green color, the melanosomes of the melanophores disperse and finally surround the iridophores and partly the xanthophores. The iridophores change from cup-shape to a cylindrical or conical shape with a simultaneous change in the orientation of the platelets from being parallel to the upper surface of the iridophores to being more irregular. The xanthophores change from lens-shape to plate-shape. The color change from green to grey seems always to go through a transitional black-green or dark olive green to dark grey. During this change the xanthophores migrate down between the iridophores, and in grey skins they are sometimes found beneath them. The pterinosomes gather in the periphery of the cell, while the carotenoid vesicles aggregate around the nucleus. The iridophores in grey skin are almost ball-shaped with concentric layers of platelets. A lighter grey color arises from a darker grey by an aggregation of melanosomes. The chromatophore values previously defined for Hyla cinerea are applicable in Hyla arborea, and the ultrastructural studies support the assumptions previously made to explain these values.The author wishes to thank Drs. P. Budtz, J. Dyck and L.O. Larsen for valuable discussions and J. Dyck for kindly providing the spectrophotometer granted him by the Danish National Science Foundation. The skilled technical assistance of Mrs. E. Schiøtt Hansen is gratefully acknowledged. Permission was granted by the Springer-Verlag to republish the illustrations of W.J. Schmidt (1920)     

A new pathway in the photoreaction cycle of trans-bacteriorhodopsin and the absorption spectra of its intermediates

The intermediates of trans-bacteriorhodopsin (trans-bR) in the photoreaction cycle were investigated under two different conditions. In a low salt and neutral pH medium (10 mM phosphate buffer, pH 6.6), trans-bR was irradiated with 500 nm light at –190 C, resulting in formation of batho-trans-bR (batho-bR^t). On warming in the dark, batho-bR^t converted to lumi-trans-bR (lumi-bR^t), meta-trans-bR (meta-bR^t) and finally to trans-bR. The intermediates N and O, which had been detected by others by flash photolysis, were not observed. The thermal decay of lumi-bR^t in a high salt and high pH medium (10 mM borate buffer with l M NaCl, pH 10.0) proceeded simultaneously through two pathways; one to meta-bR^t and another to trans-bR. About 72% of lumi-bR^t converted to trans-bR directly and the residue converted to meta-bR^t. By use of this value, the absorption spectra of batho-bR^t (_max: 626 nm), lumi-bR^t (_max: 543 nm) and meta-bR^t (_max: 418 nm) were calculated. A photoreaction cycle of bacteriorhodopsin was proposed on the basis of the above findings.     

Folin-Ciocalteu分光光度法测定夏枯草总酚含量的研究(英文)

通过采用Folin-Ciocalteu试剂,以没食子酸为对照品,用紫外可见分光光度法,研究测定夏枯草中总酚含量。结果表明,在装有样品的10 mL容量瓶中依次加入Folin-Ciocalteu试剂0.5 mL,20%Na2CO31.7 mL,室温放置60 min后,在波长660 nm或760 nm测定吸光度。多酚质量浓度在0～10.3μg/mL范围内与吸光度有良好的线性关系,回归方程在660 nm为Y=94.542X-0.0067,R2=0.9999,加样回收率为101.7～105.0%;760 nm为Y=101.13X-0.0191,R2=0.9990,加样回收率为105.1～107.6%。本研究在稳定性、准确性和重复性方面都具有较好的实验结果,可为夏枯草多酚的定量分析提供方法参考。     

测定芦荟素和芦荟大黄素的硼砂光度法

利用硼砂与芦荟中两种主要有效成分的不同作用将可见分光光度法和荧光分光光度法结合提出了测定芦荟素和芦荟大黄素含量的新方法.芦荟素的测定下限为0.00004g/L;芦荟大黄素的测定下限为0.0005g/L.此方法重复性好,样品分析结果相对标准偏差小于7%,加标回收率92～110%.     

虎杖营养器官蒽醌类化合物含量的季节变化

采用差示分光光度法测定虎杖营养器官中蒽醌类化合物的含量.该法以醋酸镁甲醇液为显色剂,以大黄素作标准对照品.其回归方程y=0.0576x+0.0012,r=0.9962.结果表明,在一个生长期中,各营养器官总蒽醌的变化不明显,蒽醌类化合物在各营养器官中含量的顺序依次为根>叶>茎;三年生的根高于一年生根;幼嫩茎叶的含量高于成熟的茎叶.在三年生根中游离蒽醌的含量在7月和9月出现两个峰值,8月降至最低;一年生根、幼叶、幼茎和老茎中游离蒽醌的含量在7月左右最高;除根外,幼叶的蒽醌类化合物的含量较其它营养器官高,且其生物量大,因此幼叶具有一定的开发前景.     

Development of 99mTc radiolabeled A85380 derivatives targeting cerebral nicotinic acetylcholine receptor: Novel radiopharmaceutical ligand 99mTc-A-YN-IDA-C4

Nicotinic acetylcholine receptors (nAChRs) are pentameric ligand-gated ion channels that have been implicated in higher brain functions. To elucidate the functional mechanisms underlying nAChRs and contribute significantly to development of drugs targeting neurological and neuropsychiatric diseases, non-invasive nuclear medical imaging can be used for evaluation. In addition, technetium-99m (^99mTc) is a versatile radionuclide used clinically as a tracer in single-photon emission computed tomography. Because A85380 is known as a potent α4β2-nAChR agonist, we prepared A85380 derivatives labeled with ^99mTc using a bifunctional chelate system. A computational scientific approach was used to design the probe efficiently. We used non-radioactive rhenium (Re) for a ^99mTc analog and found that one of the derivatives, Re-A-YN-IDA-C4, exhibited high binding affinity at α4β2-nAChR in both the docking simulation (?19.3 kcal/mol) and binding assay (Ki = 0.4 ± 0.04 nM). Further, ^99mTc-A-YN-IDA-C4 was synthesized using microwaves, and its properties were examined. Consequently, we found that ^99mTc-A-YN-IDA-C4, with a structure optimized by using computational chemistry techniques, maintained affinity and selectivity for nAChR in vitro and possessed efficient characteristics as a nuclear medicine molecular imaging probe, demonstrated usefulness of computational scientific approach for molecular improvement strategy.     

Skin colour in north Indian populations

The skin colour of six endogamous groups of north India has been studied reflectometrically. The percentage reflectance at upper arm and forchead of 650 adult males (20–25 years) is described and discussed using 601 (425 nm), 605 (545 nm) and 609 (685 nm) filters of the “EEL” spectrophotometer. The inter-group heterogeneity is revealed by variance-ratio test. The t-test has been applied to study inter-group differences in pigmentation. Some correspondence has been found in skin colour with caste hierarchy, which holds only when the populations living in the same region are compared. These differences are assigned to adaptive adjustments superimposed by caste endogamy and assortative mating for skin colour.     

Proteomic analysis of injured spinal cord tissue proteins using 2-DE and MALDI-TOF MS

Spinal cord injury (SCI) induces a progressive pathophysiology affecting cell survival and neurological integrity via complex and evolving molecular cascades whose interrelationships are not fully understood. Acute injury to the spinal cord undergoes sequential pathological change including hemorrhage, edema, axonal and neuronal necrosis, and demyelination. In the present study, we aimed to establish the proteomic profiles and characterization of the total protein expressed in traumatic injured spinal cord tissue by using 2-DE and matrix assisted laser desorption/ionization-TOF MS (MALDI-TOF MS). We performed proteomic analysis using 2-DE and MS to describe total proteins and differential proteins expression between normal and traumatic injured spinal cord tissues. The study discovered 947 total proteins and analyzed 219 and 270 proteins from normal and injured tissue, respectively. After 24 h of traumatic damage induction, the injured spinal cord tissue up-regulated over 39 proteins including neurofilament light chain, annexin 5, heat shock protein, tubulin beta, peripherin, glial fibrillary acidic protein delta, peroxiredoxin 2, and apolipoprotein A. Twenty-one proteins showed reduction. The majority of the modulated proteins belonged to the 13 functional categories. Proteins that were identified with neural functional category in injured tissue were considered most likely to be involved in wound healing response coupled with neurogenesis and gliogenesis.