Alteration of Blood Flow in a Venular Network by Infusion of Dextran 500: Evaluation with a Laser Speckle Contrast Imaging System

This study examined the effect of dextran-induced RBC aggregation on the venular flow in microvasculature. We utilized the laser speckle contrast imaging (LSCI) as a wide-field imaging technique to visualize the flow distribution in venules influenced by abnormally elevated levels of RBC aggregation at a network-scale level, which was unprecedented in previous studies. RBC aggregation in rats was induced by infusing Dextran 500. To elucidate the impact of RBC aggregation on microvascular perfusion, blood flow in the venular network of a rat cremaster muscle was analyzed with a stepwise reduction of the arterial pressure (100 → 30 mmHg). The LSCI analysis revealed a substantial decrease in the functional vascular density after the infusion of dextran. The relative decrease in flow velocity after dextran infusion was notably pronounced at low arterial pressures. Whole blood viscosity measurements implied that the reduction in venular flow with dextran infusion could be due to the elevation of medium viscosity in high shear conditions (> 45 s^-1). In contrast, further augmentation to the flow reduction at low arterial pressures could be attributed to the formation of RBC aggregates (< 45 s^-1). This study confirmed that RBC aggregation could play a dominant role in modulating microvascular perfusion, particularly in the venular networks.     

Monitoring Hypoxia Induced Changes in Cochlear Blood Flow and Hemoglobin Concentration Using a Combined Dual-Wavelength Laser Speckle Contrast Imaging and Doppler Optical Microangiography System

A synchronized dual-wavelength laser speckle contrast imaging (DWLSCI) system and a Doppler optical microangiography (DOMAG) system was developed to determine several ischemic parameters in the cochlea due to a systemic hypoxic challenge. DWLSCI can obtain two-dimensional data, and was used to determine the relative changes in cochlear blood flow, and change in the concentrations of oxyhemoglobin (HbO), deoxyhemoglobin (Hb) and total hemoglobin (HbT) in mice. DOMAG can obtain three-dimensional data, and was used to determine the changes in cochlear blood flow with single vessel resolution. It was demonstrated that during a hypoxic challenge there was an increase in the concentrations of Hb, a decrease in the concentrations of HbO and cochlear blood flow, and a slight decrease in the concentration of HbT. Also, the rate of change in the concentrations of Hb and HbO was quantified during and after the hypoxic challenge. The ability to simultaneously measure these ischemic parameters with high spatio-temporal resolution will allow the detailed quantitative analysis of several hearing disorders, and will be useful for diagnosing and developing treatments.     

Reproducibility of Non-Invasive Assessment of Skin Endothelial Function Using Laser Doppler Flowmetry and Laser Speckle Contrast Imaging

Background

Endothelial dysfunction precedes atherosclerosis. Vasodilation induced by acetylcholine (ACh) is a specific test of endothelial function. Reproducibility of laser techniques such as laser-Doppler-flowmetry (LDF) and Laser-speckle-contrast-imaging (LSCI) to detect ACh vasodilation is debated and results expressions lack standardization. We aimed to study at a 7-day interval (i) the inter-subject reproducibility, (ii) the intra-subjects reproducibility, and (iii) the effect of the results expressions over variability.

Methods and Results

Using LDF and LSCI simultaneously, we performed two different ACh-iontophoresis protocols. The maximal ACh vasodilation (peak-ACh) was expressed as absolute or normalized flow or conductance values. Inter-subject reproducibility was expressed as coefficient of variation (inter-CV,%). Intra-subject reproducibility was expressed as within subject coefficients of variation (intra-CV,%), and intra-class correlation coefficients (ICC). Fifteen healthy subjects were included. The inter-subject reproducibility of peak-ACh depended upon the expression of the results and ranged from 55% to 162% for LDF and from 17% to 83% for LSCI. The intra-subject reproducibility (intra-CV/ICC) of peak-ACh was reduced when assessed with LSCI compared to LDF no matter how the results were expressed and whatever the protocol used. The highest intra-subject reproducibility was found using LSCI. It was 18.7%/0.87 for a single current stimulation (expressed as cutaneous vascular conductance) and 11.4%/0.61 for multiple current stimulations (expressed as absolute value).

Conclusion

ACh-iontophoresis coupled with LSCI is a promising test to assess endothelial function because it is reproducible, safe, and non-invasive. N°: {"type":"clinical-trial","attrs":{"text":"NCT01664572","term_id":"NCT01664572"}}NCT01664572.     

电针对小鼠肝脏血流灌注量影响的激光散斑成像显示

目的：应用激光散斑成像技术连续监测电针过程小鼠肝脏表面血流灌注图像,研究电针不同穴位对肝脏血流灌注量的影响,探讨激光散斑技术在针灸效应研究中的应用价值。方法：采用Moor．FLPI激光散斑血流成像系统分别对足三里组、曲泉组、非经非穴组正常小鼠电针30min以及不电针对照组连续观察30min过程中肝脏表面血流灌注量变化进行观察,分析电针不同穴位、各个时点肝脏血流变化的规律。结果：（1）肝脏激光散斑图显示电针后各电针组肝脏表面整体血流灌注均增加,肝门附近区域灌注量增加幅度大于肝脏边缘区域;（2）电针各时点各电针组肝脏血流灌注量均出现增加,电针0～20min灌注量增加率为足三里组〉曲泉组〉非经非穴组;电针25～30min为足三里组〉非经非穴组〉曲泉组。结论：激光散斑血流成像技术能够精确记录显示电针过程肝脏表面的微循环变化情况,电针可以增强正常小鼠肝脏血流灌注量,电针增加肝脏血流灌注的效应存在穴位特异性。     

Synchronized Renal Blood Flow Dynamics Mapped with Wavelet Analysis of Laser Speckle Flowmetry Data

Full-field laser speckle microscopy provides real-time imaging of superficial blood flow rate. Here we apply continuous wavelet transform to time series of speckle-estimated blood flow from each pixel of the images to map synchronous patterns in instantaneous frequency and phase on the surface of rat kidneys. The regulatory mechanism in the renal microcirculation generates oscillations in arterial blood flow at several characteristic frequencies. Our approach to laser speckle image processing allows detection of frequency and phase entrainments, visualization of their patterns, and estimation of the extent of synchronization in renal cortex dynamics.     

Contrast Enhanced Ultrasound Imaging for Assessment of Spinal Cord Blood Flow in Experimental Spinal Cord Injury

Reduced spinal cord blood flow (SCBF) (i.e., ischemia) plays a key role in traumatic spinal cord injury (SCI) pathophysiology and is accordingly an important target for neuroprotective therapies. Although several techniques have been described to assess SCBF, they all have significant limitations. To overcome the latter, we propose the use of real-time contrast enhanced ultrasound imaging (CEU). Here we describe the application of this technique in a rat contusion model of SCI. A jugular catheter is first implanted for the repeated injection of contrast agent, a sodium chloride solution of sulphur hexafluoride encapsulated microbubbles. The spine is then stabilized with a custom-made 3D-frame and the spinal cord dura mater is exposed by a laminectomy at ThIX-ThXII. The ultrasound probe is then positioned at the posterior aspect of the dura mater (coated with ultrasound gel). To assess baseline SCBF, a single intravenous injection (400 µl) of contrast agent is applied to record its passage through the intact spinal cord microvasculature. A weight-drop device is subsequently used to generate a reproducible experimental contusion model of SCI. Contrast agent is re-injected 15 min following the injury to assess post-SCI SCBF changes. CEU allows for real time and in-vivo assessment of SCBF changes following SCI. In the uninjured animal, ultrasound imaging showed uneven blood flow along the intact spinal cord. Furthermore, 15 min post-SCI, there was critical ischemia at the level of the epicenter while SCBF remained preserved in the more remote intact areas. In the regions adjacent to the epicenter (both rostral and caudal), SCBF was significantly reduced. This corresponds to the previously described “ischemic penumbra zone”. This tool is of major interest for assessing the effects of therapies aimed at limiting ischemia and the resulting tissue necrosis subsequent to SCI.     

How to Build a Multiscale Model in Biology

Biological processes span several scales in space, from the single molecules to organisms and ecosystems. Multiscale modelling approaches in biology are useful to take into account the complex interactions between different organisation levels in those systems. We review several single- and multiscale models, from the most simple to the complex ones, and discuss their properties from a multiscale point of view. Approaches based on master equations for stochastic processes, individual-based models, hybrid continuous-discrete models and structured PDE models are presented.     

Application of Thinned-Skull Cranial Window to Mouse Cerebral Blood Flow Imaging Using Optical Microangiography

In vivo imaging of mouse brain vasculature typically requires applying skull window opening techniques: open-skull cranial window or thinned-skull cranial window. We report non-invasive 3D in vivo cerebral blood flow imaging of C57/BL mouse by the use of ultra-high sensitive optical microangiography (UHS-OMAG) and Doppler optical microangiography (DOMAG) techniques to evaluate two cranial window types based on their procedures and ability to visualize surface pial vessel dynamics. Application of the thinned-skull technique is found to be effective in achieving high quality images for pial vessels for short-term imaging, and has advantages over the open-skull technique in available imaging area, surgical efficiency, and cerebral environment preservation. In summary, thinned-skull cranial window serves as a promising tool in studying hemodynamics in pial microvasculature using OMAG or other OCT blood flow imaging modalities.     

A Method for 2-Photon Imaging of Blood Flow in the Neocortex through a Cranial Window

The ability to image the cerebral vasculature (from large vessels to capillaries) and record blood flow dynamics in the intact brain of living rodents is a powerful technique. Using in vivo 2-photon microscopy through a cranial window it is possible to image fluorescent dyes injected intravenously. This permits one to image the cortical vasculature and also to obtain measurements of blood flow. This technique was originally developed by David Kleinfeld and Winfried Denk. The method can be used to study blood flow dynamics during or after cerebral ischemia, in neurodegenerative disorders, in brain tumors, or in normal brain physiology. For example, it has been used to study how stroke causes shifts in blood flow direction and changes in red blood cell velocity or flux in and around the infarct. Here we demonstrate how to use 2-photon microscopy to image blood flow dynamics in the neocortex of living mice using fluorescent dyes injected into the tail vein.Download video file.^{(39M, mov)}     

532nm 激光血管内照射血液实验方法

根据现有的低强度５３２ｎｍ激光器的输出特性,建立了简便、可靠的激光血管内照射血液实验方法。通过选择合适的滤光片,可以确保５３２ｎｍ激光的单纯性;通过分束监视,实现了激光输出功率的准确计量;在实验过程中,根据监视功率值对激光输出功率进行实时调节,可以使其比较稳定地保持在设定附近。该实验方法亦可供其它激光生物效应实验参考。     

How to Build a Standardized Country-Specific Environmental Food Database for Nutritional Epidemiology Studies

There is a lack of standardized country-specific environmental data to combine with nutritional and dietary data for assessing the environmental impact of individual diets in epidemiology surveys, which are consequently reliant on environmental food datasets based on values retrieved from a heterogeneous literature. The aim of this study was to compare and assess the relative strengths and limits of a database of food greenhouse gas emissions (GHGE) values estimated with a hybrid method combining input/output and LCA approaches, with a dataset of GHGE values retrieved from the literature. France is the geographical perimeter considered in this study, but the methodology could be applied to other countries. The GHGE of 402 foodstuffs, representative of French diet, were estimated using the hybrid method. In parallel, the GHGE of individual foods were collected from existing literature. Median per-food-category GHGE values from the hybrid method and the reviewed literature were found to correlate strongly (Spearman correlation was 0.83), showing similar rankings of food categories. Median values were significantly different for only 5 (out of 29) food categories, including the ruminant meats category for which the hybrid method gave lower estimates than those from existing literature. Analysis also revealed that literature values came from heterogeneous studies that were not always sourced and that were conducted under different LCA modeling hypotheses. In contrast, the hybrid method helps build reliably-sourced, representative national standards for product-based datasets. We anticipate this hybrid method to be a starting point for better environmental impact assessments of diets.     

Know Your Enemy: How to Build and Vanquish a Global Fungal Scourge

The 8th International Conference on Cryptococcus and Cryptococcosis, chaired by Maurizio Del Poeta (Medical University of South Carolina), and organized together with June Kwon-Chung (National Institute of Allergy and Infectious Diseases), Stuart Levitz (University of Massachusetts Medical School), and John Perfect (Duke University), occurred in May 2011. This meeting brought together the world's leading researchers on Cryptococcus and cryptococcosis, including basic scientists, epidemiologists, and clinicians, to discuss new developments in Cryptococcus biology. With more than 60 oral presentations and 180 posters, this meeting enhanced our understanding of pathogenicity of Cryptococcus and served as a robust forum that facilitated cross-disciplinary discussions, research, and clinical collaborations. Due to space constraints, this brief overview highlights only a few of the topics discussed in this meeting, focusing on the evolution of virulence, host and pathogen interactions, fungal and host signaling, new advances of genomics studies on Cryptococcus, and the current status of the outbreak caused by C. gattii. The 8th International Conference on Cryptococcus and Cryptococcosis brought together scientists from across the globe in the beautiful historical downtown setting of Charleston to share their latest findings and highlight advances in Cryptococcus research. With more than 250 participants, this meeting was the largest gathering of the Cryptococcus international community in the 24-year history. Here, we review the advances presented and the current state of knowledge in the field.     

MITICS (MALDI Imaging Team Imaging Computing System): a new open source mass spectrometry imaging software

MITICS is a new software developed for MALDI imaging. We tried to render this software compatible with all types of instruments. MITICS is divided in two parts: MITICS control for data acquisition and MITICS Image for data processing and images reconstruction. MITICS control is available for Applied BioSystems MALDI-TOF instruments and MITICS Image for both Applied BioSystems and Bruker Daltonics ones. MITICS Control provides an interface to the user for setting the acquisition parameters for the imaging sequence, namely set instruments acquisition parameters, create the raster of acquisition and control post-acquisition data processing, and provide this settings to the automatic acquisition software of the MALDI instrument. MITICS Image ensures image reconstruction, files are first converted to XML files before being loaded in a database. In MITICS image we have chosen to implement different data representations and calculations for image reconstruction. MITICS Image uses three different representations that have shown to ease extraction of information from the whole data set. It also offers image reconstruction base either on the maximum peak intensity or the peak area. Image reconstruction is possible for single ions but also by summing signals of different ions. MITICS was validated on biological cases.     

Usefulness of Speckle Tracking Imaging to Assess Myocardial Contractility in Intra-Abdominal Hypertension: Study in a Mini-Pig Model

This study evaluated the usefulness of speckle tracking imaging (STI) in assessment of myocardial contractility in intra-abdominal hypertension experimentally induced in mini-pigs. To this effect, 12 mini-pigs were anesthetized with intravenous injection of 3?% sodium pentobarbital, hemorrhaged to reach the shock status, and resuscitated with excessive volume of lactated Ringer??s solution. The animals were either sham-operated (study group 1) or underwent treatment with intra-abdominal volume increment (study group 2). Observations were made prior to induction of shock, 1?h after shock, 2?h after induction of intra-abdominal hypertension, and 8 and 12?h after treatment. The heart rate and mean artery pressure were conventionally measured. STI was used to assess radial and circumferential strains of segmental ventricular wall. The results obtained demonstrated that myocardial contractility, as manifested by radial and circumferential strains of different ventricular wall segments, was decreased after induction of intra-abdominal hypertension. Treatment with intra-abdominal volume increment was able to decrease heart rate and intra-bladder pressure (indicator of effectiveness of treatment) and significantly improved myocardial contractility of involved ventricular wall segments. In conclusion, STI is a useful method to assess myocardial regional functions.     

Imaging by Delayed Light Emission (Phytoluminography) as a Method for Detecting Damage to the Photosynthetic System

An improved apparatus for obtaining luminescence (delayed light emission) images of plants is described. It consists of a phosphoroscope equipped with an imaging lens and an electronic image intensifier. It is also equipped with light-sources for obtaining images with reflected light and fluorescence light. It is shown that damage to the photosynthetic system caused by virus, insects, high or low temperature, ultraviolet radiation, or herbicide, and also chioroplast senescence as part of a normal developmental process, can be followed by this non-destructive method. In many cases changes which are not visible in fluorescence images are clearly seen in luminescence images.     

Application of a Microcomputer-Based System to Control and Monitor Bacterial Growth

A modular microcomputer-based system was developed to control and monitor various modes of bacterial growth. The control system was composed of an Apple II Plus microcomputer with 64-kilobyte random-access memory; a Cyborg ISAAC model 91A multichannel analog-to-digital and digital-to-analog converter; paired MRR-1 pH, pO₂, and foam control units; and in-house-designed relay, servo control, and turbidimetry systems. To demonstrate the flexibility of the system, we grew bacteria under various computer-controlled and monitored modes of growth, including batch, turbidostat, and chemostat systems. The Apple-ISAAC system was programmed in Labsoft BASIC (extended Applesoft) with an average control program using ca. 6 to 8 kilobytes of memory and up to 30 kilobytes for datum arrays. This modular microcomputer-based control system was easily coupled to laboratory scale fermentors for a variety of fermentations.     

Use of an Implantable Loop Recorder in a Chimpanzee (Pan troglodytes) to Monitor Cardiac Arrhythmias and Assess the Effects of Acupuncture and Laser Therapy

Cardiovascular disease is a leading cause of death in captive chimpanzees and is often associated with myocardial fibrosis, which increases the risk of cardiac arrhythmias. In this case report, we present a 36-y-old male chimpanzee (Pan troglodytes) diagnosed with frequent ventricular premature complexes (VPC). We placed a subcutaneous implantable loop recorder for continual ECG monitoring to assess his arrhythmias without the confounding effects of anesthetics. During his initial treatment with the antiarrhythmia medication amiodarone, he developed thrombocytopenia, and the drug was discontinued. After reviewing other potential therapies for the treatment of cardiac arrhythmias, we elected to try acupuncture and laser therapy in view of the positive results and the lack of adverse side effects reported in humans. We used 2 well-known cardiac acupuncture sites on the wrist, PC6 (pericardium 6) and HT7 (heart 7), and evaluated the results of the therapy by using the ECG recordings from the implantable loop recorder. Although periodic increases in the animal''s excitement level introduced confounding variables that caused some variation in the data, acupuncture and laser therapy appeared to decrease the mean number of VPC/min in this chimpanzee.Abbreviations: HT7, acupuncture site heart 7; ILR: implantable loop recorder; PC6: acupuncture site pericardium 6; VPC, ventricular premature complexesCardiovascular disease is one of the leading causes of death in captive chimpanzees (Pan troglodytes).^9,15 This incidence represents a similarity to humans, in whom cardiovascular disease is also the leading cause of death,⁸ although the pathologic processes differ between these species. Although human heart disease generally results from coronary artery atherosclerosis, heart disease in chimpanzees is associated with a diffuse interstitial myocardial fibrosis;³¹ however, both processes can be fatal. A survey of 87 deceased adult chimpanzees found evidence of cardiac disease in 68% of the cases, among which it was the primary cause of death in 34%.²⁶ At another NHP facility, 38% (n = 13) of all deaths reported in a 6-y period were associated with sudden cardiac death.¹⁴ All of these animals had been diagnosed with various degrees of cardiomyopathy prior to death, and 12 of the 13 cases were diagnosed with interstitial myocardial fibrosis according to histology.¹⁴Myocardial fibrosis is correlated with an increased risk of cardiac arrhythmias^2,7 and is a common cause of sudden cardiac death in chimpanzees, occurring much more frequently in male than in female chimps.^12,29,31 In chimpanzees, interstitial myocardial fibrosis has a random distribution throughout the myocardium. As the fibrosis progresses, it is thought to disrupt cardiac electrical signals, potentially leading to a fatal arrhythmia that presents as a sudden cardiac death.³¹ The mechanism underlying interstitial myocardial fibrosis is not entirely understood but might be related to age-associated activation of fibroblasts and diminished reparative stimuli.¹ In addition, inflammation might play a role in the development of interstitial myocardial fibrosis, given that cardiac fibrosis and remodeling are common sequelae to inflammation.¹¹ Other factors that increase the risk of developing cardiac arrhythmias in chimpanzees include age (older than 30 y), male sex, and structural heart disease.^4,12Given the high rate of mortality caused by cardiovascular disease in chimpanzees, detecting the disease early and beginning appropriate therapeutics are important. Electrocardiography is a simple diagnostic test that can detect cardiac rhythm abnormalities. However, anesthesia can serve as a confounding variable during ECG analysis of sedated chimpanzees. For the safety of both people and animals during physical examinations, chimpanzees are sedated by using tiletamine–zolazepam; anesthesia is then maintained with isoflurane. These anesthetic agents can cause decreases in heart rate, blood pressure, and cardiac output, as well as respiratory depression.²⁰ In humans, the inhalant agent isoflurane prolongs the QT interval, and this prolonged cardiac repolarization can cause ventricular arrhythmias.²⁸ Given the limited application of tiletamine–zolazepam, which primarily is used as a veterinary drug, no information regarding the association of tiletamine–zolazepam with cardiac arrhythmias in humans is available. However, cardiac arrhythmias are not uncommon in sedated chimpanzees, in which the most common arrhythmia is ventricular premature complexes (VPC).^4,27 Although VPC can occur in sedated chimpanzees with no apparent adverse effects, multiform VPC are associated with the highest risk of mortality.⁴Acknowledging the confounding effects of anesthesia in evaluation of ECG and cardiac arrhythmias, assessing ECG data from a nonsedated chimpanzee likely would be beneficial. In this report, we present a case in which we subcutaneously implanted a cardiac loop recorder to obtain ECG data longitudinally from a nonsedated chimpanzee. This technique was used successfully previously to investigate cardiac arrhythmias in 4 adult chimpanzees.¹³ Here we present the ECG findings from a male chimpanzee with cardiac arrhythmia and describe the effects of both antiarrhythmic pharmaceutical agents and the use of integrative medicine in treating the arrhythmia.     

Laser Impact on Marine Planktonic Diatoms: An Experimental Study Using a Flow Cytometry System

A flow cytometry system was used to evaluate the impact of pulsed laser irradiations from an Nd:YAG laser on two marine coastal water diatoms, Chaetoceros gracilis and Skeletonema costatum. Three flow speeds, i.e. 9, 18 and 27 ml min^{m 1} and three laser fluences, i.e. 0.025, 0.05 and 0.1 J cm^{m 2} pulse^{m 1} were tested during this study. The reduction in cell density and chlorophyll a (chl a) concentrations were monitored by reference to non-irradiated samples as controls. Upon irradiation, the cell density and the chl a concentrations became reduced significantly compared to the control (one way ANOVA p <0.001 for the cell density in both the species and p <0.05 for chl a concentrations in both species). A maximum mortality of 0.77 log₁₀ (about 83%) for C. gracilis and 0.68 log₁₀ (about 78%) for S. costatum was observed at 9 ml min^{m 1} flow speed and 0.1 J cm^{m 2} laser fluence. The maximum reduction observed in the chl a concentration was about 26% (control 0.413 and sample 0.306 mg ml^{m 1}) for C. gracilis and 27% (control 0.222 and sample 0.16 mg ml^{m 1}) for S. costatum, when the flow rate was 9 ml min^{m 1} and the fluence 0.1 J cm^{m 2}. In general, mortality increased with an increase in the laser fluence. The results thus show if the cooling water is laser-irradiated to mitigate biofouling, this could result in significant damage to the planktonic flora of the flowing seawater system, which in turn might reduce algal biofilm formation on industrially important structures. The reduction in the chl a concentration showed that the laser irradiations also could result in a significant reduction in the primary productivity of the cooling water.     

How to (or not to) communicate science

Protagonists for 'the public understanding of science' still sometimes fail to recognize that there is also a need for 'the scientists' understanding of the public' and that for most of science most of the time we are all public. 'Science' is communicated to 'the public' through popular books, museums, TV, the Internet, but far too often the present state of scientific belief is presented uncritically as the onward march of truth as discovered by Euro-American males. This has contributed to a widespread public concern, if not mistrust, in many areas of science, not least genetics and neuroscience. Although researchers often criticize the media for misrepresenting their work, the hype and simplifications often begin with the press releases put out by the researchers, their institutions and the scientific journals themselves. I conclude by looking more optimistically at the ways in which, by bringing natural science into theatre, novels and other art forms, the fragmentation of our culture may be diminished.