Thoracic Hemisection in Rats Results in Initial Recovery Followed by a Late Decrement in Locomotor Movements,with Changes in Coordination Correlated with Serotonergic Innervation of the Ventral Horn

Lateral thoracic hemisection of the rodent spinal cord is a popular model of spinal cord injury, in which the effects of various treatments, designed to encourage locomotor recovery, are tested. Nevertheless, there are still inconsistencies in the literature concerning the details of spontaneous locomotor recovery after such lesions, and there is a lack of data concerning the quality of locomotion over a long time span after the lesion. In this study, we aimed to address some of these issues. In our experiments, locomotor recovery was assessed using EMG and CatWalk recordings and analysis. Our results showed that after hemisection there was paralysis in both hindlimbs, followed by a substantial recovery of locomotor movements, but even at the peak of recovery, which occurred about 4 weeks after the lesion, some deficits of locomotion remained present. The parameters that were abnormal included abduction, interlimb coordination and speed of locomotion. Locomotor performance was stable for several weeks, but about 3–4 months after hemisection secondary locomotor impairment was observed with changes in parameters, such as speed of locomotion, interlimb coordination, base of hindlimb support, hindlimb abduction and relative foot print distance. Histological analysis of serotonergic innervation at the lumbar ventral horn below hemisection revealed a limited restoration of serotonergic fibers on the ipsilateral side of the spinal cord, while on the contralateral side of the spinal cord it returned to normal. In addition, the length of these fibers on both sides of the spinal cord correlated with inter- and intralimb coordination. In contrast to data reported in the literature, our results show there is not full locomotor recovery after spinal cord hemisection. Secondary deterioration of certain locomotor functions occurs with time in hemisected rats, and locomotor recovery appears partly associated with reinnervation of spinal circuitry by serotonergic fibers.     

Diagnosis of Clinical Bovine Mastitis by Fine Needle Aspiration Followed by Staining and Scanning Electron Microscopy in a Prototheca zopfii Outbreak

Fruit anthracnose of ugurassa caused by Colletotrichum acutatum is hereby reported for the first time in Sri Lanka and it is proposed that C. acutatum is considered together with C. gloeosporioides, as a causal agent of this disease. C. acutatum was characterised by fusiform conidia and white to orange colonies with slight shades of light mouse grey aerial mycelia. C. gloeosporioides produced grey colonies with a dark mouse grey centre and conidia were cylindrical. The other differences between the ugurassa isolate of C. acutatum and C. gloeosporioides were slower growth at temperatures ranging from 15-30 degrees C and extremely high tolerance of two fungicides, carbendazim and thiophanate methyl.     

A flavoenzyme model: Facile oxidation of thiols by a flavin immobilized in cationic polyelectrolytes

The reactions of a polymer-bound flavin with thiols (2-mercaptoethanol, glutathione, thiophenol, and 1,4-butanedithiol) are remarkably accelerated, when compared with that of a monomeric flavin. The rate enhancements observed were 30- to 6000-fold. In particular, thiophenol which had been believed not to be oxidized by flavin in nonenzymatic systems was oxidized most rapidly among the monothiols examined. The reaction rates were improved by incorporation of a dodecyl group into the flavin-containing polymer. Therefore, the hydrophobic nature of the cationic polymer matrix was concluded to be responsible for the large rate enhancement among other factors.     

Morphologic Characteristics of Choroid in the Major Choroidal Thickening Diseases,Studied by Optical Coherence Tomography

We investigated morphologic features of choroid in the choroidal thickening diseases, including central serous chorioretinopathy (CSC), polypoidal choroidal vasculopathy (PCV), and Vogt-Koyanagi-Harada disease (VKH), by a novel tomographic classification system of the choroid. This cross-sectional study involved 30 patients with active CSC, 30 patients with active PCV, and 27 patients with active VKH, and 30 normal controls. Utilizing enhanced depth imaging optical coherence tomography, we classified the morphology of the choroid into five categories: 1) Standard (S), 2) Dilated outer layer and Attenuated inner layer (DA), 3) Darkened (D), 4) Marbled (M), and 5) Pauci-Vascular (PV) types. Additional tomographic characteristics of the choroid such as choroidal vascular dilation, convolution, scleral invisibility, and choroidal hyper- or hypo-thickening were identified as well. The distribution of five choroidal tomographic morphology and additional tomographic characteristics in each group were analyzed. The DA type was observed in the CSC group more frequently than in the normal control group (53.3% vs 3.3%, P < 0.001). Additional tomographic characteristics, such as choroidal vascular dilation (76.7%), and choroidal hyper-thickening (36.7%), were more prevalent in the CSC group than in the control group. The PCV group showed higher prevalence of DA type (33.3% vs. 3.3%, P = 0.006) than the control group. The VKH group showed a significantly higher frequency of the D type (63.0%), convolution (40.7%), and scleral invisibility (70.4%) than controls (0% for all three findings). In conclusion, CSC and PCV shared common morphologic characteristics of choroid, including dilated outer vascular layer and focally attenuated innermost layer. Dense hypo-reflectivity and convolution of choroid were the specific tomographic markers for acute VKH. A new tomographic classification system of choroid may provide discrimination ability and insight into major pachychoroidopathies.     

Spectral absorption by screening pigment granules in the compound eyes of a moth and a wasp

Summary The spectral absorption by single ommin containing pigment granules or clusters of granules from compound eyes was measured spectrophotometrically between 300 and 700 nm. The measurements were made on fresh and fixed slices from compound eyes of Celerio euphorbiae and Vespa spec. In the visible part of the spectrum there is an absorption maximum between 540 and 550 nm, situated nearly 30 nm more towards the red than that of pure ommin in solution. A frequently found side maximum of variable height at about 450 nm is probably caused by oxidized xanthommatin occurring additionally within the granules. The absorption increases from 350 nm towards shorter wavelengths, and gradually declines between 550 and 700 nm.This work was supported partly by the Swedish Medical Research Council, Stiftelsen Gustaf och Tyra Svenssons Minne, Reservationsanslaget, and partly by the Air Force Office of Scientific Research through the European Office of Aerospace Research, OAR, United States Air Force under grant number EOOAR-68-0036.     

Investigation of Retinal Morphology Alterations Using Spectral Domain Optical Coherence Tomography in a Mouse Model of Retinal Branch and Central Retinal Vein Occlusion

Retinal vein occlusion is a leading cause of visual impairment. Experimental models of this condition based on laser photocoagulation of retinal veins have been described and extensively exploited in mammals and larger rodents such as the rat. However, few reports exist on the use of this paradigm in the mouse. The objective of this study was to investigate a model of branch and central retinal vein occlusion in the mouse and characterize in vivo longitudinal retinal morphology alterations using spectral domain optical coherence tomography. Retinal veins were experimentally occluded using laser photocoagulation after intravenous application of Rose Bengal, a photo-activator dye enhancing thrombus formation. Depending on the number of veins occluded, variable amounts of capillary dropout were seen on fluorescein angiography. Vascular endothelial growth factor levels were markedly elevated early and peaked at day one. Retinal thickness measurements with spectral domain optical coherence tomography showed significant swelling (p<0.001) compared to baseline, followed by gradual thinning plateauing two weeks after the experimental intervention (p<0.001). Histological findings at day seven correlated with spectral domain optical coherence tomography imaging. The inner layers were predominantly affected by degeneration with the outer nuclear layer and the photoreceptor outer segments largely preserved. The application of this retinal vein occlusion model in the mouse carries several advantages over its use in other larger species, such as access to a vast range of genetically modified animals. Retinal changes after experimental retinal vein occlusion in this mouse model can be non-invasively quantified by spectral domain optical coherence tomography, and may be used to monitor effects of potential therapeutic interventions.     

Spectral Characteristics of Phragmites australis and Its Response to Riverine Nitrogen and Phosphorus Contents in River Reaches Restored by Reclaimed Water

再生水是城市景观河湖的重要补给水源, 然而再生水中含量较高的氮和磷营养盐会引起水体富营养化, 破坏水生态平衡。以再生水补给的潮白河为研究区, 运用高光谱技术分析了挺水植物芦苇(Phragmites australis)叶片的光谱特征, 并结合水质数据, 通过拟合模型, 探究了芦苇对再生水中氮和磷的响应关系。结果表明, 各采样点水体的总氮(TN)和总磷(TP)含量分别介于1.85-18.16 mg·L^-1及0.01-0.36 mg·L^-1之间, 叶绿素a (Chl a)和溶解氧(DO)含量的范围分别为0.60-47.45 μg·L^-1与4.24-11.4 mg·L^-1。水体富营养化较为严重, 但仍处于富氧环境。多重方差分析表明, 不同采样点之间水体的TN、TP和Chl a含量差异显著(P<0.05)。由光谱反射率及反射率一阶导数曲线可知, 水体TN含量越高, 叶片光谱在可见光区的反射率越小, 红边位置也越向波长长的方向移动(即红移)。相关分析表明, 水体TN和TP含量与吸光度值log(1/R)在可见光区的相关性较强, 且TN与log(1/R)的相关系数高于TP。芦苇叶片光谱可在一定程度上区分水体TN含量差异, 但TP对光谱特征的影响模式不明显。光谱指数与水体TN含量之间的拟合模型中, 基于光化学指数(PRI)、修正叶绿素吸收指数(MCARI)和导数叶绿素指数(DCI)的模型能够解释水体TN含量变化的62.4%-70.9% (P<0.05), 可用于再生水氮含量的定量监测。该研究证明了植物光谱技术在水体富营养化监测上的可行性, 为保障再生水修复河道水质和生态安全提供了科学依据。     

再生水补给河道内芦苇的光谱特征及其对水体氮和磷含量的响应

再生水是城市景观河湖的重要补给水源, 然而再生水中含量较高的氮和磷营养盐会引起水体富营养化, 破坏水生态平衡。以再生水补给的潮白河为研究区, 运用高光谱技术分析了挺水植物芦苇(Phragmites australis)叶片的光谱特征, 并结合水质数据, 通过拟合模型, 探究了芦苇对再生水中氮和磷的响应关系。结果表明, 各采样点水体的总氮(TN)和总磷(TP)含量分别介于1.85-18.16 mg·L^-1及0.01-0.36 mg·L^-1之间, 叶绿素a (Chl a)和溶解氧(DO)含量的范围分别为0.60-47.45 μg·L^-1与4.24-11.4 mg·L^-1。水体富营养化较为严重, 但仍处于富氧环境。多重方差分析表明, 不同采样点之间水体的TN、TP和Chl a含量差异显著(P<0.05)。由光谱反射率及反射率一阶导数曲线可知, 水体TN含量越高, 叶片光谱在可见光区的反射率越小, 红边位置也越向波长长的方向移动(即红移)。相关分析表明, 水体TN和TP含量与吸光度值log(1/R)在可见光区的相关性较强, 且TN与log(1/R)的相关系数高于TP。芦苇叶片光谱可在一定程度上区分水体TN含量差异, 但TP对光谱特征的影响模式不明显。光谱指数与水体TN含量之间的拟合模型中, 基于光化学指数(PRI)、修正叶绿素吸收指数(MCARI)和导数叶绿素指数(DCI)的模型能够解释水体TN含量变化的62.4%-70.9% (P<0.05), 可用于再生水氮含量的定量监测。该研究证明了植物光谱技术在水体富营养化监测上的可行性, 为保障再生水修复河道水质和生态安全提供了科学依据。     

Optical measurement of aqueous potassium concentration by a hydrophobic indicator in lipid vesicles

An assay was developed for K+ in aqueous solution at neutral pH. The method was based on the change in optical absorbance of the hydrophobic indicator 7-(n-decyl)-2-methyl-4-(3',5'-dichlorophen-4'-one)indonaphthl++ +-1-ol (MEDPIN) in phospholipid vesicles. Formation of a ternary complex between a valinomycin-K+ pair and the anionic form of MEDPIN in the bilayer resulted in an absorption band at 584 nm. K+ concentration was determined by monitoring the MEDPIN absorbance at 584 nm and MEDPIN quenching of lissamine rhodamine B sulfonylphosphatidylethanolamine (L-RhB-PE) fluorescence by an energy-transfer mechanism. Both the fluorescence intensity and lifetime of L-RhB-PE decreased by more than 25% upon addition of 50 mM K+. Kinetic studies using stopped-flow photometry showed a single-exponential reaction of MEDPIN and valinomycin in vesicles with aqueous K+ (maximum rate 1.7 s-1) that was dependent upon [valinomycin] and [K+]. The lipid surface charge was shown to influence the ratio of anionic to neutral MEDPIN at constant pH, and to alter the sensitivity of MEDPIN absorbance to aqueous [K+]. A 1:20 neutral/negative lipid mole ratio was optimal for K+ detection at pH 7.4. Spectroscopic and kinetic data suggest that the optical response of MEDPIN to K+ involves the formation of a ternary complex between K+, valinomycin and MEDPIN.     

Andes Hantavirus-Infection of a 3D Human Lung Tissue Model Reveals a Late Peak in Progeny Virus Production Followed by Increased Levels of Proinflammatory Cytokines and VEGF-A

Andes virus (ANDV) causes hantavirus pulmonary syndrome (HPS), a severe acute disease with a 40% case fatality rate. Humans are infected via inhalation, and the lungs are severely affected during HPS, but little is known regarding the effects of ANDV-infection of the lung. Using a 3-dimensional air-exposed organotypic human lung tissue model, we analyzed progeny virus production and cytokine-responses after ANDV-infection. After a 7–10 day period of low progeny virus production, a sudden peak in progeny virus levels was observed during approximately one week. This peak in ANDV-production coincided in time with activation of innate immune responses, as shown by induction of type I and III interferons and ISG56. After the peak in ANDV production a low, but stable, level of ANDV progeny was observed until 39 days after infection. Compared to uninfected models, ANDV caused long-term elevated levels of eotaxin-1, IL-6, IL-8, IP-10, and VEGF-A that peaked 20–25 days after infection, i.e., after the observed peak in progeny virus production. Notably, eotaxin-1 was only detected in supernatants from infected models. In conclusion, these findings suggest that ANDV replication in lung tissue elicits a late proinflammatory immune response with possible long-term effects on the local lung cytokine milieu. The change from an innate to a proinflammatory response might be important for the transition from initial asymptomatic infection to severe clinical disease, HPS.     

Optical resolution of racemic 2-hydroxy octanoic acid by lipase-catalyzed hydrolysis in a biphasic membrane reactor

Racemic 2-hydroxy octanoic acid methyl ester was optically resolved by lipase-catalyzed hydrolysis in a biphasic membrane reactor using hydrophilic/hydrophobic capillary membranes. In a buffer/hexane biphasic membrane reactor using hydrophilic ultrafiltration membranes, (S)-2-hydroxy octanoic acid was recovered from the aqueous phase at 59–67% yield and 0.9–0.92 enantiomeric excess (ee), and the ester of (R)-isomer was recovered from the organic phase at 73–75% yield and 0.92–0.99 ee.     

Characteristics of protein splicing in trans mediated by a semisynthetic split intein

Protein splicing in trans results in the ligation of two protein or peptide segments linked to appropriate intein fragments. We have characterized the trans-splicing reaction mediated by a naturally expressed, approximately 100-residue N-terminal fragment of the Mycobacterium tuberculosis intein and a synthetic peptide containing the 38 C-terminal intein residues, and found that the splicing reaction was very versatile and robust. The efficiency of splicing was nearly independent of temperature between 4 and 37 degrees C and pH between 6.0 and 7.5, with only a slight decline at pH values as high as 8.5. In addition, there was considerable flexibility in the choice of the C-terminal intein fragment, no significant difference in protein ligation efficiency being observed between reactions utilizing the N-terminal fragment and either the naturally expressed 107-residue C-terminal portion of the intein, much smaller synthetic peptides, or the 107-residue C-terminal intein fragment modified by fusion of a maltose binding protein domain to its N-terminus. The ability to use different types of the C-terminal intein fragments and a broad range of reaction conditions make protein splicing in trans a versatile tool for protein ligation.     

Characteristics of mutagenesis induced by nitrosomethylurea in a mammalian multilocus system in vitro

The mutagenic effect of short- and long-term MNU exposition causing alkylation and that combined with carbamoylation, correspondingly, at the four specific gene loci of the CHO-AT3-2 Chinese hamster line was studied. The increase both in MNU mutagenic effects and in the range of induced changes resulted from intensification of carbamoylating processes. True point mutations occurred mainly on alkylation. When carbamoylation is increased, the quantity and variety of mutations change towards accumulation of the number of other genetic lesions, for example, frameshift mutations and deletions.     

Optical properties of a plasma produced by the tunneling ionization of atoms of a matter in the field of a circularly polarized wave

General features of the absorption and reflection of a test wave by a nonequilibrium plasma produced in the tunneling ionization of atoms of a matter by a circularly polarized laser pulse are described. Because of the highly anisotropic distribution of photoelectrons, the optical properties of a nonequilibrium plasma differ considerably from those of a plasma with a Maxwellian electron velocity distribution. Physically, an anomalous behavior of the absorption coefficient and of the phase shift stems from the fact that electron kinetics in the skin layer is modified by the alternating magnetic field of the test wave.     

Metabolic Characteristics of a Glycogen-Accumulating Organism in Defluviicoccus Cluster II Revealed by Comparative Genomics

Glycogen-accumulating organisms (GAOs) may compete with phosphate-accumulating organisms (PAOs) for short-chain fatty acids (VFAs) in anaerobic polyhydroxyalkanoates (PHA) synthesis, but no consequently aerobic polyphosphate accumulation in enhanced biological phosphorus removal (EBPR) process, thus deteriorating the EBPR process. They are detected frequently in the deteriorated EBPR process, but their metabolisms are still far from our comprehensions for there is seldom pure culture. In this study, a nearly complete draft genome of a GAOs in Defluviicoccus cluster II, GAO-HK, is recruited from the metagenome of activated sludge in a full-scale industrial anoxic/aerobic wastewater plant. Comparative genomics reveal similar metabolisms of PHA and glycogen in GAOs of GAO-HK, Defluviicoccus tetraformis TFO71 (TFO71) and Competibacter phosphatis clade IIA (CPIIA), and PAOs of Accumulibacter clade IIA UW-1 (UW-1) and Tetrasphaera elongata Lp2 (Lp2). Although there are similar gene cassettes related with polyphosphate metabolism in these GAOs and PAOs, especially for Defluviicoccus-relative bacteria and UW-1, ppk1 in GAOs are diverse from those in the identified PAOs, implying the difference of polyphosphate metabolism in GAOs and PAOs. Additionally, genes related to the dissimilatory denitrification are absent in TFO71 and GAO-HK, implying that additional nitrate or nitrite may favor PAOs over Defluviicoccus-relative GAOs. Therefore, PAOs suffering from competition of Defluviicoccus-relative GAOs might be rescued with the additional nitrate/nitrite, which is important to improve the stability of EBPR processes.     

人宫颈癌细胞Hela移植模型肿瘤生长的荧光分析和卡尺测量的比较

目的建立稳定表达绿色荧光蛋白的人宫颈癌细胞系,建立移植瘤模型并比较移植模型肿瘤生长的荧光分析和卡尺测量的优缺点。方法以Lipofectamine 2000介导chickenβ-actin-GFP-NEO转染人宫颈癌细胞Hela,经梯度浓度G418筛选获得稳定表达绿色荧光蛋白的细胞克隆并扩大培养。BALB/cA-nu裸鼠皮下接种1×10^6个发光细胞使其成瘤,利用活体荧光成像系统和游标卡尺观察肿瘤的生长情况。结果获得了稳定表达GFP的人宫颈癌细胞株,将其接种到裸鼠体内可成瘤。活体荧光成像观察发现,1至3周随着肿瘤体积逐渐增大,平均荧光光子数逐渐增加;4周时随着肿瘤出现明显坏死,平均荧光光子数呈现下降趋势,而游标卡尺测量结果显示肿瘤在4至5周仍然不断的增大。结论绿色荧光蛋白能够在人宫颈癌细胞Hela中长期稳定表达,用绿色荧光蛋白标记的人宫颈癌细胞Hela建立的裸鼠肿瘤模型可以为人宫颈癌研究提供理想的实验材料,应用小动物活体成像系统能够客观定量评价活的肿瘤细胞在动物体内的生长情况,而不是肿瘤体积的变化。     

Spectral broadening of the Soret band in myoglobin: an interpretation by the full spectrum of low-frequency modes from a normal modes analysis

In this work the temperature dependence of the Soret band line shape in carbon-monoxy myoglobin is re-analyzed by using both the full correlator approach in the time domain and the frequency domain approach. The new analyses exploit the full density of vibrational states of carbon-monoxy myoglobin available from normal modes analysis, and avoid the artificial division of the entire set of vibrational modes coupled to the Soret transition into "high-frequency" and "low-frequency" subsets; the frequency domain analysis, however, makes use of the so-called short-times approximation, while the time domain one avoids it. Time domain and frequency domain analyses give very similar results, thus supporting the applicability of the short-times approximation to the analysis of hemeprotein spectra; in particular, they clearly indicate the presence of spectral heterogeneity in the Soret band of carbon-monoxy myoglobin. The analyses also show that a temperature dependence of the Gaussian width parameter steeper than the hyperbolic cotangent law predicted by the Einstein harmonic oscillator and/or a temperature dependence of inhomogeneous broadening are not sufficient to obtain quantitative information on the magnitude of an-harmonic contributions to the iron-heme plane motion. However, the dependence of the previous two quantities may be used to obtain semiquantitative information on the overall coupling of the Soret transition to the low-frequency modes and therefore on the dynamic properties of the heme pocket in different states of the protein.