ABSTRACT
BACKGROUND: Ninety percent of cervical cancer (CC) diagnoses and deaths occur in low and middle-income countries (LMICs). Especially in these countries, where human and material resources are limited, there is a need for real-time screening methods that enable immediate treatment decisions (i.e., 'see and treat'). OBJECTIVE: To evaluate whether handheld vital microscopy (HVM) enables real-time detection of microvascular alterations associated with cervical intraepithelial neoplasia (CIN) and CC. METHODS: A cross-sectional study was conducted in an oncologic hospital and outpatient clinic, and included ten healthy controls, ten women with CIN, and ten women with CC. The microvasculature was assessed in four quadrants of the uterine cervix using HVM. The primary outcome was the presence of abnormal angioarchitecture (AA). Secondary outcomes included capillary loop density (CD), total vessel density (TVD), functional capillary density (FCD), and the proportion of perfused vessels (PPV). RESULTS: 198 image sequences of the cervical microvasculature were recorded. Compared to healthy controls, significantly more abnormal image sequences were observed in women with high-grade CIN (11 % vs. 44 %, P < 0.001) and women with CC (11 % vs. 69 %, P < 0.001). TVD, FCD, and PPV were lower in women with CIN and CC. CONCLUSIONS: HVM enables easy, real-time, non-invasive assessment of cervical lesions through the detection of microvascular alterations. Thereby, HVM potentially provides an opportunity for point-of-care screening, which may enable immediate treatment decisions (see and treat) and reduce the number of unnecessary surgical interventions.
Subject(s)
Uterine Cervical Dysplasia , Uterine Cervical Neoplasms , Female , Humans , Uterine Cervical Neoplasms/diagnosis , Uterine Cervical Neoplasms/pathology , Microscopy , Cross-Sectional Studies , Microcirculation , Uterine Cervical Dysplasia/diagnosis , Uterine Cervical Dysplasia/pathology , Uterine Cervical Dysplasia/surgeryABSTRACT
The goal of hemodynamic resuscitation is to optimize the microcirculation of organs to meet their oxygen and metabolic needs. Clinicians are currently blind to what is happening in the microcirculation of organs, which prevents them from achieving an additional degree of individualization of the hemodynamic resuscitation at tissue level. Indeed, clinicians never know whether optimization of the microcirculation and tissue oxygenation is actually achieved after macrovascular hemodynamic optimization. The challenge for the future is to have noninvasive, easy-to-use equipment that allows reliable assessment and immediate quantitative analysis of the microcirculation at the bedside. There are different methods for assessing the microcirculation at the bedside; all have strengths and challenges. The use of automated analysis and the future possibility of introducing artificial intelligence into analysis software could eliminate observer bias and provide guidance on microvascular-targeted treatment options. In addition, to gain caregiver confidence and support for the need to monitor the microcirculation, it is necessary to demonstrate that incorporating microcirculation analysis into the reasoning guiding hemodynamic resuscitation prevents organ dysfunction and improves the outcome of critically ill patients.
Subject(s)
Critical Care , Microcirculation , Resuscitation , Critical Care/trends , Hemodynamics , Artificial IntelligenceABSTRACT
BACKGROUND: The effect of thoracic epidural analgesia (TEA) on splanchnic blood flow during abdominal surgery remains unclear. The purpose of this study was to examine whether the hemodynamic effects of TEA resulted in microcirculatory alterations to the intestinal serosa, which was visualized using incident dark-field (IDF) videomicroscopy. METHODS: An observational cohort study was performed. In 18 patients, the microcirculation of the intestinal serosa was visualized with IDF. Microcirculatory and hemodynamic measurements were performed prior to (T1) and after administering a bolus of levobupivacaine (T2). If correction of blood pressure was indicated, a third measurement was performed (T3). The following microcirculatory parameters were calculated: microvascular flow index, proportion of perfused vessels, perfused vessel density and total vessel density. Data are presented as median [IQR]. RESULTS: Mean arterial pressure decreased from 73 mmHg (68-83) at T1 to 63 mmHg (±11) at T2 (p = 0.001) with a systolic blood pressure of 114 mmHg (98-128) and 87 (81-97), respectively (p = 0.001). The microcirculatory parameters of the bowel serosa, however, were unaltered. In seven patients, blood pressure was corrected to baseline values from a MAP of 56 mmHg (55-57), while microcirculatory parameters remained constant. CONCLUSION: We examined the effects of TEA on the intestinal serosal microcirculation during abdominal surgery using IDF imaging for the first time in patients. Regardless of a marked decrease in hemodynamics, microcirculatory parameters of the bowel serosa were not significantly affected. TRIAL REGISTRY NUMBER: ClinicalTrials.gov identifier NCT02688946.
Subject(s)
Anesthesia, Epidural , Anesthetics, Local , Intestine, Small/physiopathology , Levobupivacaine , Microcirculation , Serous Membrane/physiopathology , Aged , Anesthetics, Local/pharmacology , Arterial Pressure/drug effects , Cohort Studies , Female , Humans , Intestine, Small/blood supply , Intestine, Small/diagnostic imaging , Levobupivacaine/pharmacology , Male , Microcirculation/drug effects , Microscopy, Video , Middle Aged , Serous Membrane/blood supply , Serous Membrane/diagnostic imaging , Thoracic VertebraeABSTRACT
BACKGROUND: Recognition of a non-viable bowel during colorectal surgery is a challenging task for surgeons. Identifying the turning point in serosal microcirculatory deterioration leading up to a non-viable bowel is crucial. The aim of the present study was to determine whether sidestream darkfield (SDF) imaging can detect subtle changes in serosal microcirculation of the sigmoid after vascular transection during colorectal surgery. METHODS: A prospective observational clinical study was performed at a single medical centre. All eligible participants underwent laparoscopic sigmoid resection and measurements were taken during the extra-abdominal phase. Microcirculation was measured at the transected bowel and 20 cm proximal to this point. Microcirculatory parameters such as Microvascular Flow Index (MFI), proportion of perfused vessels (PPV), perfused vessel density (PVD), total vessel density (TVD) and the Heterogeneity Index were determined. Data are presented as median (interquartile range) or mean ± standard deviation. RESULTS: A total of 60 SDF images were acquired for 10 patients. Perfusion parameters and perfused vessel density were significantly lower at the transected bowel compared with the non-transected measurements [MFI 2.29 (1.96-2.63) vs 2.96 (2.73-3.00), p = 0.007; PPV 74% (55-83) vs 94% (86-97), p = 0.007; and PVD 7.61 ± 2.99 mm/mm2 versus 10.67 ± 1.48 mm/mm2, p = 0.009]. Total vessel density was similar between the measurement locations. CONCLUSIONS: SDF imaging can identify changes of the bowel serosal microcirculation. Significantly lower serosal microcirculatory parameters of the vascular transected bowel was seen compared with the non-transected bowel. The ability of SDF imaging to detect subtle differences holds promise for future research on microvascular cut-off values leading to a non-viable bowel.
Subject(s)
Colon, Sigmoid/surgery , Diagnostic Techniques, Cardiovascular , Intraoperative Care/methods , Serous Membrane/blood supply , Serous Membrane/diagnostic imaging , Aged , Colon, Sigmoid/blood supply , Feasibility Studies , Female , Humans , Male , Microcirculation/physiology , Middle Aged , Prospective StudiesABSTRACT
Microcirculatory alterations play a pivotal role in sepsis-related morbidity and mortality. However, since the microcirculation has been a "black box", current hemodynamic management of septic patients is still guided by macrocirculatory parameters. In the last decades, the development of several technologies has shed some light on microcirculatory evaluation and monitoring, and the possibility of incorporating microcirculatory variables to clinical practice no longer seems to be beyond reach. The present review provides a brief summary of the current technologies for microcirculatory evaluation, and attempts to explore the potential role and benefits of their integration to the resuscitation process in critically ill septic patients.
Subject(s)
Blood Gas Analysis/methods , Microcirculation , Microscopy, Video , Monitoring, Physiologic , Point-of-Care Systems , Sepsis/physiopathology , Arterioles/physiopathology , Blood Gas Analysis/instrumentation , Capillaries/physiopathology , Carbon Dioxide/blood , Hemodynamics , Humans , Hypoxia/physiopathology , Oximetry/instrumentation , Oximetry/methods , Oxygen/blood , Partial Pressure , Resuscitation , Sepsis/blood , Spectroscopy, Near-Infrared/instrumentation , Spectroscopy, Near-Infrared/methodsABSTRACT
AIMS: Vascularisation of the vagina is necessary for optimal function and support of the surrounding organs. We evaluated whether vaginal microcirculation, as representative of vascularisation, differs between women with and without pelvic organ prolapse (POP). METHODS: In 17 women with POP-Q stage ≥2 and 10 women without POP measurements were performed using sidestream dark-field imaging (groups were not matched). POP and non-POP sites were compared in women with a single compartment prolapse (n = 7). Morphology of the microvessels was scored using the microcirculatory architecture and capillary tortuosity scores at four regions of the vaginal wall. Capillary density measurements were performed and microvascular flow was assessed according to the microvascular flow index (MFI) score. RESULTS: Architecture and tortuosity scores were similar for each anatomical region between women with and without POP and between the POP and non-POP site. A statistically significant difference in capillary density in the left vaginal wall between women with and without POP was observed (25.8 vs. 34.0, P = 0.049). No significant differences in capillary density were observed between the POP and non-POP site. Average MFI scores revealed a continuous flow for all four regions in patients with and without POP. CONCLUSIONS: Vaginal microcirculatory architecture, capillary density, and microvascular flow are similar in women with and without POP. Our method to assess the microcirculation of the vaginal wall is consistent in women with POP, which generates an opportunity to assess vaginal microcirculation in the (surgical) treatments of POP. Neurourol. Urodynam. 35:764-770, 2016. © 2015 Wiley Periodicals, Inc.
Subject(s)
Microcirculation/physiology , Microvessels/physiopathology , Pelvic Organ Prolapse/physiopathology , Vagina/blood supply , Adult , Aged , Aged, 80 and over , Female , Humans , Middle Aged , Vagina/physiopathologyABSTRACT
In neonates, cardiovascular system development does not stop after the transition from intra-uterine to extra-uterine life and is not limited to the macrocirculation. The microcirculation (MC), which is essential for oxygen, nutrient, and drug delivery to tissues and cells, also develops. Developmental changes in the microcirculatory structure continue to occur during the initial weeks of life in healthy neonates. The physiologic hallmarks of neonates and developing children make them particularly vulnerable during critical illness; however, the cardiovascular monitoring possibilities are limited compared with critically ill adult patients. Therefore, the development of non-invasive methods for monitoring the MC is necessary in pediatric critical care for early identification of impending deterioration and to enable the initiation and titration of therapy to ensure cell survival. To date, the MC may be non-invasively monitored at the bedside using hand-held videomicroscopy, which provides useful information regarding the microcirculation. There is an increasing number of studies on the MC in neonates and pediatric patients; however, additional steps are necessary to transition MC monitoring from bench to bedside. The recently introduced concept of hemodynamic coherence describes the relationship between changes in the MC and macrocirculation. The loss of hemodynamic coherence may result in a depressed MC despite an improvement in the macrocirculation, which represents a condition associated with adverse outcomes. In the pediatric intensive care unit, the concept of hemodynamic coherence may function as a framework to develop microcirculatory measurements towards implementation in daily clinical practice.
Subject(s)
Critical Illness/therapy , Hemodynamic Monitoring/trends , Microcirculation/physiology , Pediatrics/methods , Hemodynamic Monitoring/methods , Hemodynamics/physiology , Humans , Infant , Infant, Newborn , Intensive Care Units, Neonatal/organization & administration , Pediatrics/standardsABSTRACT
BACKGROUND: Lactate, acetate, and gluconate are anions used in balanced resuscitation fluids, of which lactate and acetate are considered bicarbonate precursors. This study investigated the role of the liver in the ability of balanced and unbalanced solutions to correct acid-base alterations and renal haemodynamics and microvascular oxygenation in a rat model of resuscitated haemorrhagic shock. METHODS: Ringer's lactate, Ringer's acetate, PlasmaLyte, or normal saline were administered following haemorrhagic shock in the presence or absence of a 70% partial liver resection. Renal haemodynamics and microvascular oxygenation (by oxygen-dependent quenching of phosphorescence) were measured as well as concentrations of lactate, gluconate, and acetate in plasma and urine. Kidney wet and dry weight was also assessed. RESULTS: Partial liver resection resulted in increased liver enzymes compared with control and shock groups (P < 0.01). Haemorrhagic shock decreased systemic and renal perfusion and reduced microvascular kidney oxygenation with lactic acidosis (P < 0.01). Resuscitation with balanced fluids did not fully restore renal oxygenation (P < 0.01). Ringer's acetate and PlasmaLyte increased bicarbonate content and restored pH better than Ringer's lactate or saline after partial liver resection (P < 0.01). Liver resection caused an increase in plasma gluconate after PlasmaLyte resuscitation (P < 0.05). CONCLUSIONS: Acetate-buffered balanced fluids show superior buffering effects compared with Ringer's lactate or saline. Gluconate is partially metabolized by the liver, although it does not contribute to acid-base control because of its excretion in urine. Acetate is metabolized regardless of liver function and may be the most efficient bicarbonate precursor. Lactate infusion tends to overwhelm the metabolism capacity of the residual liver.
Subject(s)
Bicarbonates/metabolism , Fluid Therapy , Liver/physiology , Shock, Hemorrhagic/therapy , Acetates/metabolism , Acid-Base Equilibrium , Animals , Kidney/metabolism , Lactic Acid/metabolism , Male , Rats , Rats, Sprague-Dawley , Resuscitation , Shock, Hemorrhagic/metabolismABSTRACT
Normal functioning of myocardium requires adequate oxygenation, which in turn is dependent on an adequate microcirculation. NADH-fluorimetry enables a direct evaluation of the adequacy of tissue oxygenation while the measurement of quenching of Pd-porphyrine (PpIX) phosphorescence enables quantitative measurement of microvascular pO2. Combination of these two techniques provides information about the relation between microvascular oxygen content and parenchymal oxygen availability in Langendorff hearts. In normal myocardium there is heterogeneity at the microcirculatory level resulting in the existence of microcirculatory weak units, originating at the capillary level, which reoxygenate the slowest upon reoxygenation after an episode of ischemia. Sepsis and myocardial hypertrophia alter the pattern of oxygen transport whereby the microcirculation is disturbed at the arteriolar/arterial level. NADH fluorimetry also reveals a disturbance of mitochondrial oxygen availability in sepsis. Furthermore it is shown that these techniques can also be applied to various organs and tissues in vivo.
Subject(s)
Coronary Circulation , Energy Metabolism , Mitochondria, Heart/metabolism , Animals , In Vitro Techniques , Microcirculation , Myocardium/metabolism , NAD/analysis , Oxygen/metabolism , RatsABSTRACT
Incident dark field imaging (IDF) is a new generation handheld microscope for bedside visualization and quantification of microcirculatory alterations. IDF is the technical successor of sidestream dark field imaging (SDF), currently the most used device for microcirculatory measurements. In (pre)term neonates the reduced thickness of the skin allows non-invasive transcutaneous measurements. The goal of this study was to compare the existing device (SDF) and its technical successor (IDF) in preterm neonates. We hypothesized that IDF imaging produces higher quality images resulting in a higher vessel density. After written informed consent was given by the parents, skin microcirculation was consecutively measured on the inner upper arm with de SDF and IDF device. Images were exported and analyzed offline using existing software (AVA 3.0). Vessel density and perfusion were calculated using the total vessel density (TVD) proportion of perfused vessels (PPV) and perfused vessel density. The microcirculation images quality score was used to evaluate the quality of the video images. In a heterogeneous group of twenty preterm neonates (median GA 27.6 weeks, range 24-33.4) IDF imaging visualized 19.9% more vessels resulting in a significantly higher vessel density (TVD 16.9 vs. 14.1/mm, p value < 0.001). The perfusion of vessels could be determined more accurately in the IDF images, resulting in a significant lower PPV (88.7 vs. 93.9%, p value 0.002). The IDF video images scored optimal in a higher percentage compared to the SDF video images. IDF imaging of the cutaneous microcirculation in preterm neonates resulted in a higher vessel density and lower perfusion compared to the existing SDF device.
Subject(s)
Image Enhancement/instrumentation , Infant, Premature/physiology , Microcirculation/physiology , Microscopy/instrumentation , Skin Physiological Phenomena , Skin/blood supply , Blood Flow Velocity/physiology , Equipment Design , Equipment Failure Analysis , Female , Humans , Infant, Newborn , Male , Microscopy, Video/instrumentation , Microvessels/cytology , Microvessels/physiology , Reproducibility of Results , Sensitivity and SpecificityABSTRACT
BACKGROUND: Despite many clinical trials and investigative efforts to determine appropriate therapeutic intervention(s) for shock, this topic remains controversial. The use of i.v. fluid has represented the cornerstone for the treatment of hypoperfusion for two centuries. METHODS: As a part of International Acute Dialysis Quality Initiative XII Fluids Workgroup meeting, we sought to incorporate recent advances in our understanding of vascular biology into a more comprehensive yet accessible approach to the patient with hypoperfusion. In this workgroup, we attempted to develop a framework that incorporates key aspects of the vasculature into a diagnostic approach. RESULTS: The four main components of our proposal involve the assessment of the blood flow (BF), vascular content (vC), the vascular barrier (vB), and vascular tone (vT). Any significant perturbation in any of these domains can lead to hypoperfusion at both the macro- and micro-circulatory level. We have termed the BF, vC, vB, and vT diagnostic approach the vascular component (VC) approach. CONCLUSIONS: The VC approach to hypoperfusion has potential advantages to the current diagnostic system. This approach also has the distinct advantage that it can be used to assess the systemic, regional, and micro-vasculature, thereby harmonizing the approach to clinical vascular diagnostics across these levels. The VC approach will need to be tested prospectively to determine if this system can in fact improve outcomes in patients who suffer from hypoperfusion.
Subject(s)
Blood Vessels/physiopathology , Fluid Therapy/methods , Fluid Therapy/standards , Hemodynamics/physiology , Delphi Technique , Dialysis , Humans , Microcirculation , Perfusion , Regional Blood Flow/physiology , Sepsis/therapyABSTRACT
Hypoxia stimulates angiogenesis through the binding of hypoxia-inducible factors to the hypoxia-response element in the vascular endothelial growth factor (Vegf) promotor. Here, we report that deletion of the hypoxia-response element in the Vegf promotor reduced hypoxic Vegf expression in the spinal cord and caused adult-onset progressive motor neuron degeneration, reminiscent of amyotrophic lateral sclerosis. The neurodegeneration seemed to be due to reduced neural vascular perfusion. In addition, Vegf165 promoted survival of motor neurons during hypoxia through binding to Vegf receptor 2 and neuropilin 1. Acute ischemia is known to cause nonselective neuronal death. Our results indicate that chronic vascular insufficiency and, possibly, insufficient Vegf-dependent neuroprotection lead to the select degeneration of motor neurons.
Subject(s)
Cell Hypoxia/genetics , Endothelial Growth Factors/genetics , Lymphokines/genetics , Motor Neurons/pathology , Nerve Degeneration/genetics , Response Elements/genetics , Amyotrophic Lateral Sclerosis/genetics , Amyotrophic Lateral Sclerosis/pathology , Animals , Axons/physiology , Binding Sites , Electrophysiology , Endothelial Growth Factors/metabolism , Humans , Lymphokines/metabolism , Mice , Mice, Knockout , Motor Neurons/physiology , Muscle Contraction , Muscle Fibers, Skeletal/pathology , Muscular Atrophy/genetics , Muscular Atrophy/pathology , Nerve Degeneration/pathology , Nerve Degeneration/physiopathology , Nerve Tissue Proteins/genetics , Nerve Tissue Proteins/metabolism , Neuropilin-1 , Peripheral Nerves/pathology , Promoter Regions, Genetic , Receptor Protein-Tyrosine Kinases/genetics , Receptor Protein-Tyrosine Kinases/metabolism , Receptors, Growth Factor/genetics , Receptors, Growth Factor/metabolism , Receptors, Vascular Endothelial Growth Factor , Sequence Deletion , Spinal Cord/physiology , Vascular Endothelial Growth Factor A , Vascular Endothelial Growth FactorsABSTRACT
Ischemia/reperfusion injury and inflammation are associated with microcirculatory dysfunction, endothelial injury and glycocalyx degradation. This study aimed to assess microcirculation in the sublingual, intestinal and the (remnant) liver in patients undergoing major liver resection, to define microcirculatory leukocyte activation and its association with glycocalyx degradation. In this prospective observational study, the microcirculation was assessed at the beginning of surgery (T0), end of surgery (T1) and 24 h after surgery (T2) using Incident Dark Field imaging. Changes in vessel density, blood flow and leukocyte behaviour were monitored, as well as clinical parameters. Syndecan-1 levels as a parameter of glycocalyx degradation were analysed. 19 patients were included. Sublingual microcirculation showed a significant increase in the number of rolling leukocytes between T0 and T1 (1.5 [0.7-1.8] vs. 3.7 [1.7-5.4] Ls/C-PCV/4 s respectively, p = 0.001), and remained high at T2 when compared to T0 (3.8 [3-8.5] Ls/C-PCV/4 s, p = 0.006). The microvascular flow decreased at T2 (2.4 ± 0.3 vs. baseline 2.8 ± 0.2, respectively, p < 0.01). Duration of vascular inflow occlusion was associated with significantly higher numbers of sublingual microcirculatory rolling leukocytes. Syndecan-1 increased from T0 to T1 (42 [25-56] vs. 107 [86-164] ng/mL, p < 0.001). The microcirculatory perfusion was characterized by low convection capacity and high number of rolling leukocytes. The ability to sublingually monitor the rolling behaviour of the microcirculatory leukocytes allows for early identification of patients at risk of increased inflammatory response following major liver resection.
Subject(s)
Hemodynamics , Hepatectomy , Leukocytes , Liver/blood supply , Liver/surgery , Microcirculation , Time-Lapse Imaging , Aged , Biomarkers/blood , Blood Gas Analysis , Comorbidity , Female , Hepatectomy/methods , Humans , Leukocyte Count , Leukocytes/cytology , Male , Microvascular Density , Middle AgedABSTRACT
Analysis of membrane potential recordings upon microelectrode impalement of four types of macrophages (cell lines P388D1 and PU5-1.8, cultured mouse peritoneal macrophages, and cultured human monocytes) reveals that these cells have membrane potentials at least two times more negative than sustained potential values (E(s)) frequently reported. Upon microelectrode entry into the cell (P388D1), the recorded potential drops to a peak value (E(p)) (mean -37 mV for 50 cells, range -15 to -70 mV) within 2 ms, after which it decays to a depolarized potential (E(n)) (mean -12 mV) in about 20 ms. Thereafter, the membrane develops one or a series of slow hyperpolarizations before a final sustained membrane potential (E(s)) (mean -14 mV, range -5 to -40) is established. The mean value of the peak of the first hyperpolarization (E(h)) is -30 mV (range -10 to -55 mV). The initial fast peak transient, measured upon microelectrode entry, was first described and analyzed by Lassen et al. (Lassen, U.V., A.M. T. Nielson, L. Pape, and L. O. Simonsen, 1971, J. Membr. Biol. 6:269-288 for other change in the membrane potential from its real value before impalement to a sustained depolarized value. This was shown to be true for macrophages by two-electrode impalements of single cells. Values of E(p), E(n), E(h), E(s), and membrane resistance (R(m)) measured for the other macrophages were similar to those of P388D1. From these results we conclude that E(p) is a better estimate of the true membrane potential of macrophages than E(s), and that the slow hyperpolarizations upon impalement should be regarded as transient repolarizations back to the original membrane potentials. Thus, analysis of the initial fast impalement transient can be a valuable aid in the estimation of the membrane potential of various sorts of small isolated cells by microelectrodes.
Subject(s)
Macrophages/physiology , Animals , Ascitic Fluid/cytology , Cell Line , Cells, Cultured , Electric Conductivity , Humans , Membrane Potentials , Mice , Microelectrodes , Monocytes/physiologyABSTRACT
The present study has shown that changes in ionic channel currents accompany the phagocytosis of particles by mononuclear phagocytes. The patch-clamp technique in the cell-attached configuration was applied to human monocyte-derived macrophages to measure the activity of single transmembrane ionic channels in intact cells. During such measurements, IgG-opsonized and non-opsonized latex particles were offered for phagocytosis under continuous video-microscopical observation. Single particles were presented to the phagocytes at a membrane location some distance from that of the patch electrode. After a lag period following particle attachment, enhanced inward and outward time-variant single channel currents coinciding with particle engulfment were observed. On the basis of current-voltage characteristics and membrane potential measurements, the outward-directed channels were identified as K+ channels. Phagocytosis was also accompanied by slow transient changes in background membrane currents, probably due to changes in the membrane potential of the phagocytosing cell. Phagocytosis of IgG-coated latex particles differed from phagocytosis of uncoated or albumin-coated particles by a shorter lag time between particle attachment and the onset of enhanced ionic channel activity.
Subject(s)
Ion Channels/physiology , Macrophages/physiology , Phagocytosis , Electric Conductivity , Electrophysiology , Humans , In Vitro Techniques , Opsonin Proteins , Receptors, Fc/physiologyABSTRACT
Sidestream Dark Field (SDF) imaging, a stroboscopic LED ring-based imaging modality, is introduced for clinical observation of the microcirculation. SDF imaging is validated by comparison to Orthogonal Polarization Spectral imaging. Nailfold capillary diameters and red blood cell velocities were measured using both techniques and equal quantitative results were obtained. An image quality system was developed to quantitatively compare the quality of sublingually-acquired microcirculatory images using OPS and SDF imaging. Venular contrast, sharpness, and quality were shown to be comparable for OPS and SDF imaging. However, capillary contrast and quality were shown to be significantly higher using SDF imaging. Venular granularity, in addition, was more clearly observable using SDF imaging.
ABSTRACT
Endotracheal intubation in mice is necessary for experiments involving intratracheal instillation of various substances, repeated pulmonary function assessments and mechanical ventilation. Previously described methods for endotracheal intubation in mice require the use of injection anaesthesia to immobilize the animal during the intubation procedure or the use of a volatile anaesthetic prior to intubation for immobilization. With these methods, the control of anaesthetic depth during the intubation procedure is absent. We describe a method for simple and rapid intratracheal intubation in mice for mechanical ventilation, using a self-built plastic support to facilitate the intubation procedure. General anaesthesia is maintained by means of inhalation through a non-rebreathing circuit connected to the plastic support. This set-up gives the operator control of anaesthetic depth and sufficient time to perform the intubation procedure. A purpose-made laryngoscopic blade is used to facilitate the intubation tube entering the trachea. The blade of the purpose-made laryngoscope is constructed as a retraction guide and is curved for easy handling. Under direct vision, the epiglottis is gently lifted by the laryngoscopic blade while the intubation tube is pushed into the trachea. Following this novel intubation technique, we were able to mechanically ventilate mice for at least 2 h without severely disturbing blood gases. Histological evaluation of the lungs and microscopic evaluation of the trachea and larynx showed no signs of trauma related to the intubation technique or mechanical ventilation.
Subject(s)
Intubation, Intratracheal/veterinary , Mice , Animals , Blood Gas Analysis , Intubation, Intratracheal/instrumentation , Intubation, Intratracheal/methods , Lung/anatomy & histology , Male , Mice, Inbred C57BL , Respiration, Artificial/veterinarySubject(s)
Diagnostic Imaging/instrumentation , Microcirculation/pathology , Animals , Brain/blood supply , Brain/surgery , Cricetinae , Humans , Light , Photons , Skin/blood supply , Tongue/blood supplyABSTRACT
OBJECTIVE: In this study we evaluated the inter-observer agreement in the assessment of gingival capillary density using Orthogonal Polarization Spectral Imaging. METHODS: In this study gingival capillary density of 100 healthy subjects was determined by 2 independent observers. Agreement was quantified by calculation of the mean differences between the observers and the standard deviation of this difference and the limits of agreement. Reliability was quantified by means of the intraclass correlation coefficient (ICC). RESULTS: Fifty males and 50 females were included in the study. The mean age for males was 20+/-1.2 years and for females 20+/-1.4. OPS images showed remarkable good quality images of the gingival microcirculation. The interclass correlation between the 2 observers was 0.63 while the interclass correlation for the 6 measurements in observer 1 was 0.95 and 0.94 for observer 2. The mean capillary density for females in observer 1 was 83.69+/-16.4 and 83+/-16.0 in observer 2, versus 60.55+/-12.3 for observer 1 and 60.4+/-12.1 for males. The mean quantitative functional capillary density in male students was 60.48+/-10.7, compared to 83.45+/-13.5 in female students. CONCLUSIONS: OPS imaging enabled for the first time direct in vivo visualization and quantification of human functional gingival capillary density in healthy medical students. The inter-observer agreement was found to be good to fair on the quantification of gingival capillary density between the two independent observers. The intracorrelation coefficient (0.95) was excellent when assessing the reliability of one observer.
Subject(s)
Gingiva/blood supply , Image Processing, Computer-Assisted , Microcirculation , Adult , Capillaries/anatomy & histology , Humans , Microscopy, Polarization , Observer Variation , Reference Values , Sensitivity and SpecificityABSTRACT
Hypertrophic hearts contain areas of hypoperfusion which can be visualized by increased NADH surface fluorescence during in vitro perfusion without oxygen-carrying particles under constant pressure and pacing. By contrast, fluorescence remained low when non-hypertrophic hearts were used instead. When during perfusion of normal hearts the pH of the medium was lowered from 7.5 to 7.0, areas of high fluorescence appeared in a few minutes. The high fluorescent areas under conditions of cardiac hypertrophy or pH 7.0 perfusion could be reduced by addition of superoxide dismutase. It indicates that oxygen free radicals interfere with proper flow regulation in areas of low pH. Fluorescence in hypertrophic hearts also diminished during addition of albumin-bound oleate to the standard, glucose-containing, medium. This is in agreement with our earlier finding of fatty acid protection from acidosis-initiated loss of capillary flow (Biochim. Biophys. Acta, 1033 (1990) 214-218). In contrast to low concentrations of free fatty acids, high concentrations interfere with tissue oxygenation. This has been illustrated by the use of 1 mM octanoate, which after a few min caused the appearance of high fluorescent areas. We conclude that decompensation of flow in hypoperfused areas of heart, as occurs in hypertrophy, may be stimulated by acidosis and oxygen free radicals.