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PubMedDate not supplied

Laparoscopic fluorometry: a new minimally invasive tool for investigation of the intestinal microcirculation.

The aim of this study was to develop and establish a new system of laparoscopic fluorometry for the purpose of investigating the intestinal microcirculation. In 25 pigs (German Landrace, 16-25 kg body weight), ischemia was established in two segments (A, irreversible; B, reversible ischemia; C, internal control) of the small intestine by a laparoscopic technique. Microcirculation in the segments was assessed by laparoscopy at a second-look operation 24 h later by means of the fluorescence system Endoscan. The fluorescence of the three bowel segments was measured by arbitrary dye fluorescence units (DFU) 15 min after starting reperfusion, before and after injection of sodium fluorescein (NaFlu, 0.25 mg/kg body weight). The dividing line between viable and nonviable bowel tissue was established from the inflow and outflow rates of NaFlu with the aid of ROC (receiver operating characteristic) curves. The specificity and sensitivity of the new method were evaluated by correlating the results with the viability of each intestinal segment as predicted by three laparoscopically experienced surgeons and by histological examination. By means of the calculated separation sharpness (fluorescence index at 2 min >0.5, outflow factor of NaFlu at 10 min >20%), the overall predictions of intestinal viability in all 25 animals achieved a sensitivity of 93.5% and a specificity of 94.1% by laparoscopic fluorometry, versus a sensitivity of 70.8% and a specificity of 87.5% for the prediction of bowel viability by ordinary laparoscopic technique. Used as an adjunct to conventional laparoscopy, laparoscopic fluorometry brought significant gains in sensitivity and specificity in the distinction between reversible and irreversible intestinal ischemia.

Animals↗

PubMed2004

Analysis of leukocyte rolling and migration--using inhibitors in the undisturbed microcirculation of the rat mesentery--on inflammatory stimulation.

AIM: Our aim was to develop a method of migration analysis using the undisturbed microcirculation of rat mesentery, and using the new method, analyze leukocyte migration in casein-induced inflammation. METHOD: Sprague Dawley (SD) rats were injected with tumor necrosis factor (TNF) alpha, interleukin (IL)-1alpha, or casein intraperitoneally. Following this, the rats were sacrificed and the mesentery tissue removed was fixed and stained with Giemsa. The leukocytes were counted as a rolling index in the venules and as a migration index in the perivascular area. RESULTS: There was no relation between the diameter of venules and leukocyte migration. The time change curves of leukocyte activity in casein inflammation show about a 1 h difference between rolling and migration. From inhibitor experiments of casein-induced migration at 2 h, it has been suggested that selectin-related rolling is necessary. Platelet-activating factor (PAF) also appears partially involved. CONCLUSION: The improved undisturbed microcirculation method is helpful not only for rolling analysis but also in analysis of leukocyte migration. Casein inflammation analyzed using this method revealed that rolling is necessary and also suggested that partial involvement of PAF is necessary for pathogenesis of leukocyte extravasations.

Animals↗

PubMedDate not supplied

Expression of cytochrome P450-4A isoforms in the rat cremaster muscle microcirculation.

OBJECTIVE: This study sought to identify any specific cytochrome P450 (CYP450) -4A enzyme isoforms expressed in arterioles and/or the surrounding parenchymal tissue of the rat cremaster muscle. METHODS: RT-PCR was used to detect the presence of specific CYP450-4A isoforms in isolated muscle fibers and arterioles from the cremaster muscle of Sprague-Dawley rats; CYP450-4A protein expression was determined by Western blotting. RESULTS: CYP450-4A3 mRNA was expressed in isolated muscle fibers and in cremasteric arterioles, while CYP450-4A8 mRNA was expressed only in cremasteric arterioles. CYP450-4A1 and CYP450-4A2 mRNA were not expressed in arterioles and skeletal muscle cells, although all four isoforms were strongly expressed in the liver. CYP450-4A protein was detected in both the isolated muscle fibers and in the isolated arterioles. CONCLUSIONS: The present study identifies the specific pattern of cytochrome P450-4A isoform expression in arterioles and parenchymal cells of the skeletal muscle microcirculation, and supports the hypothesis that the cytochrome P-450 enzymes may play a role in the regulation of microvascular function in the skeletal muscle microcirculation.

Animals↗

PubMedDate not supplied

The microcirculation in severe malaria.

Severe malaria in humans and animals is initiated by interactions between malaria-infected cells, host blood cells (including monocytes, T cells and platelets) and endothelial cells of the microcirculation. Adhesion to vascular cells, and possible vascular obstruction in severe human disease, involves interaction between host receptors and parasite-derived proteins, such as the variant antigen Plasmodium falciparum erythrocyte membrane protein 1 (PfEMP1). Our understanding of how different PfEMP1 variants may target infected erythrocytes to specific sites, such as the placenta, is rapidly increasing. However, in most instances downstream immune-mediated inflammatory processes appear more central than parasite accumulation to development of severe malaria. Using genetically-manipulated animal models of severe malaria, key roles for CD8 T cells and mediators such as lymphotoxin in the pathogenesis of murine disease have been established. Experimental and human studies suggest vascular deposition of activated platelets may have a central role. Here, we review some recent advances in the understanding of severe malaria pathogenesis from human and animal studies, focusing on events at the level of the microcirculation, and highlight the role for activated host cells in initiating the pathology of the disease.

Animals↗

PubMed2005

Protective effects of ischemic preconditioning on the intestinal mucosal microcirculation following ischemia-reperfusion of the intestine.

OBJECTIVE: The small bowel villi are extremely sensitive to ischemia-reperfusion (IR) injury and a range of microcirculatory disturbances contribute to structural and functional changes. The aim of this study was to determine the protective effects of ischemic preconditioning (IPC) of the intestine on the mucosal villous microcirculation during IR injury of the intestine and whether heme oxygenase (HO) is involved in the protection. METHODS: Rats were allocated into 4 groups: (1) sham, (2) IR consisting of 30 min of ischemia followed by 2 h of reperfusion, (3) IPC, as in IR group, but preceded by 10 min of ischemia and 10 min of reperfusion, and (4) with administration of zinc protoporphyrin, an HO inhibitor before IPC and IR. The mucosa of an exteriorized segment of ileum was visualized. Mucosal perfusion index (MPI), red blood cell (RBC) velocity and leukocyte-endothelial interactions during reperfusion were assessed continuously using in vivo fluorescence microscopy. HO activity in the ileum was assessed at the end of the reperfusion period. RESULTS: IPC improved the MPI by 26% and the RBC velocity by 29% on comparison to IR. IR led to a 52% increase in leukocyte-endothelial interactions on comparison to IPC. The administration of zinc protoporphyrin reversed the beneficial effects of IPC. There was a two fold increase of HO activity in IPC compared to IR, whereas zinc protoporphyrin significantly reduced the HO activity. CONCLUSIONS: IPC conferred a protective effect on the villous microcirculation possibly via HO and might prove to be an effective strategy for the amelioration of IR injury.

Animals↗

PubMed2006

Targeted therapeutic applications of acoustically active microspheres in the microcirculation.

The targeted delivery of intravascular drugs and genes across the endothelial barrier with only minimal side effects remains a significant obstacle in establishing effective therapies for many pathological conditions. Recent investigations have shown that contrast agent microbubbles, which are typically used for image enhancement in diagnostic ultrasound, may also be promising tools in emergent, ultrasound-based therapies. Explorations of the bioeffects generated by ultrasound-microbubble interactions indicate that these phenomena may be exploited for clinical utility such as in the targeted revascularization of flow-deficient tissues. Moreover, development of this treatment modality may also include using ultrasound-microbubble interactions to deliver therapeutic material to tissues, and reporter genes and therapeutic agents have been successfully transferred from the microcirculation to tissue in various animal models of normal and pathological function. This article reviews the recent studies aimed at using interactions between ultrasound and contrast agent microbubbles in the microcirculation for therapeutic purposes. Furthermore, the authors present investigations involving microspheres that are of a different design compared to current microbubble contrast agents, yet are acoustically active and demonstrate potential as tools for targeted delivery. Future directions necessary to address current challenges and advance these techniques to clinical practicality are also discussed.

Animals↗

PubMedDate not supplied

Potassium channels in the peripheral microcirculation.

Vascular smooth muscle (VSM) cells, endothelial cells (EC), and pericytes that form the walls of vessels in the microcirculation express a diverse array of ion channels that play an important role in the function of these cells and the microcirculation in both health and disease. This brief review focuses on the K+ channels expressed in smooth muscle and endothelial cells in arterioles. Microvascular VSM cells express at least four different classes of K+ channels, including inward-rectifier K+ channels (Kin), ATP-sensitive K+ channels (KATP), voltage-gated K+ channels (Kv), and large conductance Ca2+-activated K+ channels (BKCa). VSM KIR participate in dilation induced by elevated extracellular K+ and may also be activated by C-type natriuretic peptide, a putative endothelium-derived hyperpolarizing factor (EDHF). Vasodilators acting through cAMP or cGMP signaling pathways in VSM may open KATP, Kv, and BKCa, causing membrane hyperpolarization and vasodilation. VSMBKc. may also be activated by epoxides of arachidonic acid (EETs) identified as EDHF in some systems. Conversely, vasoconstrictors may close KATP, Kv, and BKCa through protein kinase C, Rho-kinase, or c-Src pathways and contribute to VSM depolarization and vasoconstriction. At the same time Kv and BKCa act in a negative feedback manner to limit depolarization and prevent vasospasm. Microvascular EC express at least 5 classes of K+ channels, including small (sKCa) and intermediate(IKCa) conductance Ca2+-activated K+ channels, Kin, KATP, and Kv. Both sK and IK are opened by endothelium-dependent vasodilators that increase EC intracellular Ca2+ to cause membrane hyper-polarization that may be conducted through myoendothelial gap junctions to hyperpolarize and relax arteriolar VSM. KIR may serve to amplify sKCa- and IKCa-induced hyperpolarization and allow active transmission of hyperpolarization along EC through gap junctions. EC KIR channels may also be opened by elevated extracellular K+ and participate in K+-induced vasodilation. EC KATP channels may be activated by vasodilators as in VSM. Kv channels may provide a negative feedback mechanism to limit depolarization in some endothelial cells.

Animals↗

PubMedDate not supplied

Special edition of Microcirculation commemorating the 50th anniversary of the Microcirculatory Society, Inc.

This special edition of Microcirculation brings together the proceedings from the Symposium held in the spring of 2004 in the Natcher Auditorium on the campus of the National Institutes of Health to commemorate the 50th Anniversary of the Microcirculatory Society, Inc. Twelve brief reviews are introduced with the goals being to present a collective sense of the history of research in microcirculation, provide insight into where established topics of inquiry stand today, and to define emerging topics of investigation for future research efforts.

Animals↗

PubMedDate not supplied

Effects of ET(A) receptor antagonism on proinflammatory gene expression and microcirculation following hepatic ischemia/reperfusion.

BACKGROUND: The objective of this study was to investigate the effect of a specific endothelin(A) receptor antagonist (ET(A)-RA) on mRNA expression of genes encoding vasoactive mediators and proinflammatory cytokines and on the microhemodynamics (assessed by measurement of laser Doppler flow and tissue blood gases) following complete vascular exclusion of the porcine liver. STUDY DESIGN: Sixteen adult German landrace pigs were subjected to 120 min of warm hepatic ischemia by total vascular exclusion. To avoid portal congestion, a passive porto-femoro/jugular bypass was implanted. The animals were divided into 2 groups: the control group received saline solution and the therapy group was given the selective ET(A)-RA BSF 208075. Hepatic microcirculation was evaluated by p(O(2)) and p(CO(2)) measurement with the Paratrend sensor and by laser Doppler flow measurement. Liver tissue samples were collected 1 h after reperfusion and quantitative mRNA expression for prepro-ET-1, pro-IL-1beta, pro-IL-6, pro-TNF-alpha, eNOS was analyzed using the TaqMan system. Additionally, immunohistochemical analysis using a semiquantitative score for ET-1 was performed. Postischemic liver damage was monitored by measurement of liver enzymes and assessed by histological analysis using a semiquantitative scoring classification. RESULTS: Partial oxygen pressure in the hepatic tissue and laser Doppler flow were significantly improved in the therapy group. One hour after reperfusion, quantitative RT-PCR revealed significantly lower expression of prepro-ET-1, eNOS, pro-TNF-alpha, and pro-IL-6 in the therapy group compared to controls. Immunohistochemical analysis demonstrated significantly reduced ET-1 immunostaining after therapy. Furthermore, blockade of ET(A) receptors prevents tissue damage. CONCLUSIONS: Treatment with the selective ET(A)-RA BSF 208075 has protective effects on microcirculation after 120 min liver ischemia and reperfusion. The authors were able to show that ET(A)-RA not only affects the expression of vasoactive genes, but also decreases gene expression of proinflammatory cytokines such as TNF-alpha and IL-6.

Animals↗

PubMed2006

Capillary hemodynamics and oxygen pressures in the aging microcirculation.

Healthy aging acts to redistribute blood flow (Q) and thus O2 delivery (QO2 ) among and within the exercising muscles such that QO2 to highly oxidative muscle fibers may be compromised. Within the microcirculation of old muscles capillary hemodynamics are altered and the matching of QO2 to oxidative requirements (VO2) is impaired such that at exercise onset the microvascular O2 pressure falls below that seen in their younger counterparts. This is important because the microvascular O2 pressure denotes the sole driving force for blood-myocyte O2 transfer and any compromise may slow VO2 kinetics and reduce exercise tolerance. This review considers the microcirculatory evidence for a reduced perfusive (QO2 ) and diffusive O2 flux within aged muscle and highlights the pressing need for intravital microscopy studies of the muscle microcirculation during exercise.

Aging↗

PubMed1999

Changes in hepatic microcirculation and histomorphology in brain-dead organ donors: an experimental study in rats.

OBJECTIVE: To assess the effect of brain death on hormonal homeostasis, hepatic microcirculation, and histomorphology in organ donors. DESIGN: Prospective randomised experimental study. SETTING: Institute for Surgical Research, Germany. SUBJECTS: 12 male Sprague-Dawley rats INTERVENTIONS: 6 rats acted as controls, and 6 had brain death induced by inflation of an intracranial balloon. MAIN OUTCOME MEASURES: Mean arterial pressure, serum concentration of antidiuretic hormone (ADH), thyroxine (T4), free-T4, triiodothyronine (T3) and free-T3, bile production, intravital fluorescence microscopy and electron microscopic appearances. RESULTS: After induction of brain death mean arterial pressure rose within 5 minutes followed by significant hypotension (p < 0.01). ADH concentration was reduced (p < 0.01), as was bile production (p < 0.05). There was impaired sinusoidal perfusion and increased interaction between leucocytes and endothelium in the hepatic microvasculature. The electron microscopic analysis showed vacuolisation of hepatocytes. CONCLUSION: Macrohaemodynamics, ADH homeostasis, and the hepatic microcirculation deteriorate after brain death, which leads to histomorphological damage of hepatocytes and compromised liver function.

Animals↗

PubMed2006

Cerebral microcirculation shear stress levels determine Neisseria meningitidis attachment sites along the blood-brain barrier.

Neisseria meningitidis is a commensal bacterium of the human nasopharynx. Occasionally, this bacterium reaches the bloodstream and causes meningitis after crossing the blood-brain barrier by an unknown mechanism. An immunohistological study of a meningococcal sepsis case revealed that neisserial adhesion was restricted to capillaries located in low blood flow regions in the infected organs. This study led to the hypothesis that drag forces encountered by the meningococcus in the bloodstream determine its attachment site in vessels. We therefore investigated the ability of N. meningitidis to bind to endothelial cells in the presence of liquid flow mimicking the bloodstream with a laminar flow chamber. Strikingly, average blood flows reported for various organs strongly inhibited initial adhesion. As cerebral microcirculation is known to be highly heterogeneous, cerebral blood velocity was investigated at the level of individual vessels using intravital imaging of rat brain. In agreement with the histological study, shear stress levels compatible with meningococcal adhesion were only observed in capillaries, which exhibited transient reductions in flow. The flow chamber assay revealed that, after initial attachment, bacteria resisted high blood velocities and even multiplied, forming microcolonies resembling those observed in the septicemia case. These results argue that the combined mechanical properties of neisserial adhesion and blood microcirculation target meningococci to transiently underperfused cerebral capillaries and thus determine disease development.

Animals↗

PubMed2006

Evaluating microcirculation by pulsatile laser Doppler signal.

Laser Doppler flowmetry (LDF) is a popular method for monitoring the microcirculation, but it does not provide absolute measurements. Instead, the mean flux response or energy distribution in the frequency domain is generally compared before and after stimulus. Using the heartbeat as a trigger, we investigated whether the relation between pressure and flux can be used to discriminate different microcirculatory conditions. We propose the following three pulsatile indices for evaluating the microcirculation condition from the normalized pressure and flux segment with a synchronized-averaging method: peak delay time (PDT), pressure rise time and flux rise time (FRT). The abdominal aortic blood pressure and renal cortex flux (RCF) signals were measured in spontaneously hypertensive rats (SHR) and Wistar Kyoto rats (WKY). The mean value of the RCF did not differ between SHR and WKY. However, the PDT was longer in SHR (87.14 +/- 5.54 ms, mean +/- SD) than in WKY (76.92 +/- 2.62 ms; p < 0.001). The FRT was also longer in SHR (66.56 +/- 1.98 ms) than in WKY (58.02 +/- 1.77 ms; p < 0.001). We propose that a new dimension for comparing the LDF signals, which the results from the present study show, can be used to discriminate RCF signals that cannot be discriminated using traditional methods.

Animals↗

PubMed2000

Image correlation method for measuring blood flow velocity in microcirculation: correlation 'window' simulation and in vivo image analysis.

To elucidate the function of the microcirculation system, it is very important to know the blood flow velocity and its distribution in the microvessels. We have developed an automated system for measuring blood flow velocity in microcirculation by image correlation. The 'window' in the image correlation method is equivalent to the sensors in various other measurement methods. We performed simulations with virtual blood flow images consisting of random dots before measuring actual ones, and examined the optimum window shape and size. We found that by reducing the size of a circular window to the size of erythrocytes we could measure in vivo blood flow images with high accuracy. We recorded them with a high-speed video camera system at high temporal resolution, and measured the velocity in microvessels of normal Wistar Kyoto (WKY) and spontaneously hypertensive rats (SHR). SHR had higher blood velocity than WKY even though the vessel diameters were the same. Using this method to measure the blood flow velocity profile at the bent corner of SHR's arteriole at the heart systole, we found that erythrocytes flow faster at the inner side of the bend, so the vessel wall was exposed locally to higher shear stress in the hypertensive condition.

Animals↗

PubMed2003

Mechanistic probing of gaseous signal transduction in microcirculation.

Nitric oxide (NO) and carbon monoxide (CO) serve as activators of soluble guanylate cyclase (sGC) in vitro, and the latter serves as a microvascular relaxant for the liver, a major organ for heme oxygenase-dependent heme degradation and gas generation. Another important determinant of local sGC activities is superoxide anion, which scavenges NO and/or activates sGC directly. Altered bioavailability of the oxygen-derived species and its functional outcomes remain unknown, because information on amounts and distribution of these molecules has hardly been examined in vivo. Our recent studies provided evidence for such complex actions of multiple gases in vivo. Intravital visualization of NO in microcirculation revealed that two distinct sources, NO synthase-1 and -3, play a major role in the maintenance of NO in arteriolar and venular walls, respectively. Besides its vasorelaxing action in the hepatic microcirculation, CO could induce vasoconstriction in the resistant artery where NO is abundantly available; systemic blood pressure was elevated in transgenic mice overexpressing heme oxygenase-1 site-specifically in vascular smooth muscle cells. Such a relationship between the gases has also been demonstrated by mechanistic bioprobing of sGC function using novel monoclonal antibodies. This article aims to provide an overview of advances in visual assessment of the generation and reception of oxygen-derived gaseous mediators in vivo.

Animals↗

PubMed1997

Eicosanoids in bovine retinal microcirculation.

The vasoactive effects of the arachidonic acid-derived cyclooxygenase product, prostaglandin F2 alpha and PGH2 analogue U46619, on ex vivo perfused (25 microliters/min) bovine retinal microcirculation were assessed. First, we determined the ability of the cultured bovine retinal microvascular endothelial cells to synthesize eicosanoids from 14C-arachidonic acid. Bovine retinal microvascular endothelial cell cultures synthesized detectable amounts of PGE2, PGF2 alpha, PGI2, and PGH2 analogue; PGI2 product being slightly greater. The central retinal arteriole was cannulated and microcirculation continuously perfused with PSS with constant flow. Vasoactive compounds were applied in ten-fold concentration increments to PSS in the muscle chamber containing retinal preparation. The diameters of retinal microarterioles were measured using a closed-circuit video camera system. In ex vivo bovine retinal microvascular model studies, PGF2 alpha and PGH2 analogue U46619, caused dose-dependent vasoconstriction of all vessels, and these responses were attenuated by specific TP (TXA2) receptor antagonist, SQ 29,548, at 10(-6) M.

15-Hydroxy-11 alpha,9 alpha-(epoxymethano)prosta-5↗

PubMed2005

Low-level laser therapy accelerates collateral circulation and enhances microcirculation.

OBJECTIVE: To evaluate the efficacy of low-level laser therapy (LLLT) on collateral circulation and microcirculation if a blood vessel is occluded. BACKGROUND DATA: Investigators have attempted prostaglandin and ultrasound therapy to promote improvements in the vascular bed of deprived tissue after an injury, which may lead to occlusion of the blood vessels. MATERIALS AND METHODS: Thirty-four adult rabbits were used in this study, two of them considered 0-h reading group, while the rest were divided into two equal groups, with 16 rabbits each: control and those treated with LLLT. Each rabbit underwent two surgical operations; the medial aspect of each thigh was slit, the skin incised and the femoral artery exposed and ligated. The site of the operation in the treated group was irradiated directly following the operation and for 3 d after, one session daily for 10 min/session. The laser system used was a gallium-aluminum-arsenide (Ga-Al-As) diode laser with a wavelength of 904 nm and power of 10 mW. Blood samples collected from the femoral artery above the site of the ligation were sent for examination with high-performance liquid chromatography (HPLC) to determine the levels of adenosine, growth hormone (GH) and fibroblast growth factor (FGF). Tissue specimens collected from the site of the operation, consisting of the artery and its surrounding muscle fibers, were sent for histopathological examination to determine the fiber/capillary (F/C) ratio and capillary diameter. Blood samples and tissue specimens were collected at 4, 8, 12, 16, 20, 24, 48 and 72 h postoperatively from the animals of both groups, control and treated. RESULTS: Rapid increases in the level of adenosine, GH, and FGF occurred. The F/C ratio and capillary diameter peaked at 12-16 h; their levels declined gradually, reaching normal values 72 h after irradiation in the treated group. Numerous collateral blood vessels proliferated the area, with marked increases in the diameters of the original blood vessels. CONCLUSIONS: The results indicated that LLLT accelerated collateral circulation and enhanced microcirculation and seemed to be unique in the normalization of the functional features of the injured area, which could lead to occlusion of the regional blood vessels.

Adenosine↗

PubMed2006

Short-time effects of laser needle stimulation on the peripheral microcirculation assessed by laser Doppler spectroscopy and near-infrared spectroscopy.

OBJECTIVE: The aim of the present study was to evaluate immediate effects of a standardized laser needle stimulation over a defined acupuncture point on the microvascular blood flow and muscle oxygenation in the human forearm. BACKGROUND DATA: Recently, it has been shown that laser stimulation improves tissue perfusion. This is relevant since adequate blood supply is an important factor in the treatment of pain syndromes. METHODS: The study was designed as a randomized, double-blinded, placebo-controlled trial. Thirty-three healthy non-smoking males were randomly assigned to a control group (n = 15) with no laser irradiation and to an intervention group (n = 18) for which laser needle irradiation was performed on the right forearm at acupuncture point Pe6. Non-invasive blood flow measurements (laser Doppler spectroscopy [LDS]) were performed before, during, and after intervention. Additionally, the dynamic changes in muscle oxygenation of the m. flexor carpi ulnaris were investigated using near-infrared spectroscopy (NIRS). RESULTS: Repeated measures MANOVA demonstrated a statistically significant interaction between time and group (p = 0.034, effect size = 0.39), indicating that peripheral blood flow was influenced by laser needle application. In contrast, tissue oxygenation was not affected by the experimental treatment. CONCLUSION: It has been demonstrated that laser needle stimulation may improve peripheral microcirculation under standardized conditions, whereas tissue oxygenation remained unchanged. Further research is required to determine the influence of various parameter settings and irradiation treatments on the peripheral microcirculation. Moreover, different acupuncture points should be investigated in order to appraise the clinical effectiveness of laserneedle stimulation.

Acupuncture Points↗