Blood velocity measurement in human conjunctival vessels

The bulbar conjunctiva is one of the few areas in which blood flow in the peripheral vasculature can be directly and noninvasively observed in the human. Although extensive literature exists describing morphological changes which correlate with a variety of systemic diseases in this vasculature, little quantitative data is available on hemodynamics in either normal or abnormal states. The hemodynamic data available are primarily subjective assessments of "low flow." Approaches to place the subjective assessment on more quantitative grounds have usually been based on photographic techniques that have intrinsic inadequacies. The objective of the work reported here was to develop a system capable of providing sequential blood velocity data potentially useful for providing quantitative information on blood flow and its change in the microvessels of the human conjunctiva. The method that has evolved uses a standard Zeiss slit-lamp to image a subject's conjunctival vessels by using a 1-inch Newvicon TV camera with an electronic magnification of 2x. The video image is simultaneously recorded on a video tape recorder (VTR) to an overall system magnification of approximately 4 microm/raster line. The data acquisition phase requires approximately 5 minutes of patient time, whereas the actual determination of blood velocity in individual vessels is done offline through a modification of the dual-slit videodensimetric method. Two independently controllable video cursors are placed axially over the vessel image with the VTR in the still-frame mode. For each consecutive video field, the position of two reference points on the vessel and the position of each cursor relative to these and to each other are encoded into a computer to track the moving image caused by normal eye movement. The computer then determines new cursor coordinates to ensure a constant position within the vessel. The electrical signals obtained for each cursor site and for each video field are cross-correlated to yield the average blood velocity over the sampled time interval. The system has been calibrated in vitro from 0.2 to 2.5 mm/sec, evaluated in experimental animals, and used to measure blood velocity (0.3 to 1.5 mm/sec) in human conjunctival venules with diameters ranging from 20 to 50 microm. At this writing, blood velocity has been recorded during a period of about 3 months in the same vessel of several postmyocardial infarction patients. Thus, the method appears suitable for determining sequential changes in small vessel blood flow in patients over extended periods of time.     

Development of blood vessels in the human eyeball

Blood vessels of 167 eyeballs of human embryos and fetuses have been studied, using macro- microscopical, histological and morphometrical methods. Time of appearance and differentiation of blood vessels in the eye anlage has been determined. Topography and architecture of the vitreous body artery are described in detail, as well as the vascular tunic of the lens and the pupil membrane. Regularities and time of reversal development of the vascular formations mentioned are followed during the whole intrauterine development. The data obtained add to extend our knowledge on the role of disturbances of the normal morphogenesis of the eyeball blood bed in pathogenesis of its congenital diseases.     

Optical biosensors for real-time measurement of analytes in blood plasma

The preparation of assemblies consisting of multiple molecular layers of bovine serum albumin (BSA), monoclonal antibodies against horseradish peroxidase (anti-HRP), and monoclonal antibodies against methotrexate (anti-MTT), as well as interaction of the assemblies with human blood plasma were observed using a grating coupler and Young interferometer (YI). The assemblies could be arranged according to decreasing amounts of nonspecific deposits bound irreversibly to them from blood plasma as follows-an adsorbed antibody monolayer saturated with adsorbed BSA, antibody multilayers linked with polycations, antibodies covalently immobilized on a BSA layer densely crosslinked with glutaraldehyde (GA), slightly crosslinked BSA double layer, slightly crosslinked antibody double layers. The occurrence of human serum albumin (HSA), human fibrinogen (Fg), IgG, and IgM in the plasma deposits was studied by binding the respective antibodies. IgG, IgM, and Fg were detected in plasma deposits on the immobilized assemblies while the composition of a plasma deposit on the unmodified sensor surface reflected roughly the plasma composition containing mainly adsorbed HSA and Fg. A crosslinked anti-HRP double layer was immobilized on a waveguiding branch of YI and a similar anti-MTT double layer was immobilized on the other branch. The sensor response to blood plasma was fairly decreased owing to a compensation of the respective optical changes in the two branches, in which a similar non-specific adsorption took place. The addition of HRP or MTT to plasma induced specific responses of the corresponding branches.     

Adrenergic innervation of human mesenteric blood vessels.

A fluorescent histochemical technique has been applied to study the adrenergic innervation of human superior mesenteric arteries obtained at autopsy. Specific catecholamine fluorescence was demonstrated in the smaller branches of this artery taken from three infants and one child. No specific fluorescence was seen in arteries from three adult subjects.     

Superficial anastomoses of blood vessels in the human heart.

We investigated superficial interarterial, intra-arterial, arterio-venous and veno-venous anastomoses in 100 hearts from men and women taken at random who died at 20-85 years of age. By means of careful dissection and injection into the blood vessels of a differently stained 12% solution of vinilit in acetone, we studied the number and localization of the anastomoses and their appearance. Interarterial anastomoses, i.e. anastomoses between the ramification of the left and the right coronary artery, were detected in 33 hearts. Intra-arterial anastomoses, i.e. anastomoses between the ramifications of one and the same artery, were seen in 5 hearts. Arterio-venous anastomoses were found in 39 hearts. Veno-venous anostomoses were found in 49 hearts. The veno-venous anastomoses display a great many variations, being commonly the regular finding in human veins.     

Development of the cardiac blood vessels in staged human embryos

Serial paraffin sections (mostly stained with hematoxylin and eosin) of 52 human embryos at stages ranging from 13 to 20 (approximate ovulation age of 5-8 weeks) were examined. The first sign of definitive blood vessels was found to be localized in the apical incisure of the heart of an embryo at stage 14. Blood vessels of this kind closely resembled 'blood islands' in appearance, being composed of primitive erythroblasts surrounded by an outer layer of endothelium. At stage 16, a funnel-like invagination of the endothelium was recognized in the posterior wall of the sinus venosus. This structure was considered to represent one of the cardiac veins (probably the middle cardiac vein). A faint endothelial sprout of the left coronary artery was detected at stage 18, while the right one was observed later, at stage 19. Finally, at stage 20, both of the coronary arteries invariably existed with a covering of mesenchymal cells.     

Axial mechanical properties of fresh human cerebral blood vessels

Human cerebral blood vessels are frequently damaged in head impact, whether accidental or deliberate, resulting in intracranial bleeding. Additionally, the vasculature constitutes the support structure for the brain and, hence, plays a key role in the cranial load response. Quantification of its mechanical behavior, including limiting loads, is thus required for a proper understanding and modeling of traumatic brain injury--as well as providing substantial assistance in the development and application of preventive measures. It is believed that axial stretching is the dominant loading mode for the blood vessels, regardless of the nature of the insult. Eighteen arteries and fourteen veins were obtained from the cortical surface of the cerebral temporal lobe of patients undergoing surgery. These vessels were stretched to failure in the longitudinal direction, either quasi-statically or dynamically. The significance of specimen and experiment parameters was determined using multivariate analysis of variance (MANOVA) testing. Results demonstrate that the arteries were considerably stiffer than the veins, carrying approximately twice as much stress at failure but withstanding only half as much stretch. No significant rate dependence was measured over a strain rate range of more than four orders of magnitude (0.01 to 500 s -1).     

Morphological aspects of age-related adaptation of the arterial vessels of the human kidneys in various cardiovascular diseases

The investigation has been performed in 190 normal kidneys and at certain cardiovascular diseases (atherosclerosis of the abdominal aorta, hypertension, chronic ischemic cardiac disease and their combination). Principally similar morphological structures are mobilized in the process of the renal arterial bed adaptation both normal and at the cardio-vascular diseases studied. Transition from certain functional changes to the organic ones in the muscular tunic and in the elastic membranes of large arteries, in correlation of afferent and deferent glomerular arterioles of all cortical layers is stated, as well as adaptive rearrangement of the microvessels and of the juxtamedullary shunt.     

Isolating and defining cells to engineer human blood vessels

A great deal of attention has been recently focused on understanding the role that bone marrow-derived putative endothelial progenitor cells (EPC) may play in the process of neoangiogenesis. However, recent data indicate that many of the putative EPC populations are comprised of various haematopoietic cell subsets with proangiogenic activity, but these marrow-derived putative EPC fail to display vasculogenic activity. Rather, this property is reserved for a rare population of circulating viable endothelial cells with colony-forming cell (ECFC) ability. Indeed, human ECFC possess clonal proliferative potential, display endothelial and not haematopoietic cell surface antigens, and display in vivo vasculogenic activity when suspended in an extracellular matrix and implanted into immunodeficient mice. Furthermore, human vessels derived became integrated into the murine circulatory system and eventually were remodelled into arterial and venous vessels. Identification of this population now permits determination of optimal type I collagen matrix microenvironment into which the cells should be embedded and delivered to accelerate and even pattern number and size of blood vessels formed, in vivo. Indeed, altering physical properties of ECFC-collagen matrix implants changed numerous parameters of human blood vessel formation, in host mice. These recent discoveries may permit a strategy for patterning vascular beds for eventual tissue and organ regeneration.     

Microarchitectonics and microtopography of blood vessels of the human rectum

By means of injecting the vessels with 0.2% solution of silver nitrate after Ranvier, solutions of Indian ink and gas soot, impregnation after V.V. Kuprianov, the blood vessels of the rectal wall have been studied. The material has been obtained from 50 corpses of persons of various age, not suffering from any diseases of the gastrointestinal tract. The structural organization of the microcirculatory tract has been studied layer-by-layer, at all levels by means of atraumatic lamination of the tunics. Terminal links of the microcirculatory bed form zonal functional complexes of microvessels specific for each part of the rectum. They regularly repeat in a certain tunic and owing to this the whole organization of the microcirculatory bed acquires features of a definite polymeric structure which consists of homonomic complexes of microvessels. Their regularity, as regards their topography and quantity, definitely differs in each tunic and layer of the rectal wall. Precapillary sphincters and arteriolovenular anastomoses are revealed; they perform an active regulation of blood circulation in the organ. Diameters of the microvessels and density of the microcirculatory network have some slight fluctuations (differences) in functionally poorly active tunics of the rectal wall--the serous and submucous, especially in its rectosigmoid part. Differences of these parameters are especially expressed within the limits of the rectal ampule and its mucous and muscular tunics.     

Optical chip immunoassay for hCG in human whole blood

We report on the development of an integrated optic chip sensor for performing rapid and sensitive immunoassays with human whole blood using human chorionic gonadotropin (hCG) as the model system. The optical chip is based on the Hartman interferometer, which uses a single planar lightbeam to address multiple interferometers, each comprising a signal/reference pair of sensing regions. The binding of antigen to specific capture antibodies on the signal sensing region causes a change in the refractive index of the surface layer, which is detectable by its effect on the evanescent field of the guided lightbeam. The reference-sensing region is coated with an irrelevant antibody, which optically cancels a large fraction of the non-specific adsorption that occurs on the specific-sensing region when the sensor is tested with clinical specimens. This work extends previous experiments with buffer and human serum to measurements in undiluted whole human blood. Optical chips were stored dry after surface functionalization, and rehydrated with blood. Colloidal gold nanoparticles conjugated to a second anti-hCG monoclonal antibody were used to provide signal amplification, thereby enhancing assay sensitivity, in a one-step procedure with the gold conjugate added to the test sample immediately prior to measurement. Background signals due to non-specific binding (NSB) in blood were found to be higher than those previously reported with human serum. In addition, a high level of background signal was found with the gold conjugate, which had not been observed in experiments with either buffer or serum. Nevertheless, hCG could be detected at 0.5 ng/ml within 10 min of sample application. The sensor response was linear over the concentration range 0.5-5 ng/ml hCG, as compared with the clinically-relevant range 0.3-1.5 ng/ml. Detection at higher concentrations was affected by scattering from large amounts of bound gold nanoparticles. However, initial binding rate measurements could be used to maintain assay quantitation.     

Optical flow sensor for continuous invasive measurement of blood flow velocity

Continuous monitoring of intrapulse measurement of blood flow in humans is currently not achievable with clinically available instruments. In this paper, we demonstrate a method of measuring the instantaneous variations in flow during pulsatile blood flow with an optical flow sensor comprising a fiber Bragg grating sensor and illumination from a 565 nm Light‐Emitting‐Diode. The LED illumination heats the blood and fluctuations in temperature, due to variations in flow, are detected by the fiber sensor. A set of experiments at different flow rates (20 to 900 mL/min) are performed in a simulated cardiac circulation setup with pulsatile flow. Data are compared with an in‐line time of flight ultrasound flow sensor. Our results show that the optical and ultrasonic signals correlate with Pearson coefficients ranging from ?0.83 to ?0.98, dependent on the pulsatile frequency. Average flow determined by ultrasound and the optical fiber sensor showed a parabolic relationship with R² = 0.99. An abrupt step change in flow induced by occlusion and release of the circuit tubing demonstrated that the optical fiber and ultrasound sensor had similar response. The method described is capable of intrapulse blood flow measurement under pulsatile flow conditions, with potential applications in medicine where continuous blood flow sensing is desired.