Imaging blood vessels and lymphatic vessels in the zebrafish.

Blood vessels supply tissues and organs with oxygen, nutrients, cellular, and humoral factors, while lymphatic vessels regulate tissue fluid homeostasis, immune trafficking, and dietary fat absorption. Understanding the mechanisms of vascular morphogenesis has become a subject of intense clinical interest because of the close association of both types of vessels with pathogenesis of a broad spectrum of human diseases. The zebrafish provides a powerful animal model to study vascular morphogenesis because of their small, accessible, and transparent embryos. These unique features of zebrafish embryos permit sophisticated high-resolution live imaging of even deeply localized vessels during embryonic development and even in adult tissues. In this chapter, we summarize various methods for blood and lymphatic vessel imaging in zebrafish, including nonvital resin injection-based or dye injection-based vessel visualization, and alkaline phosphatase staining. We also provide protocols for vital imaging of vessels using microangiography or transgenic fluorescent reporter zebrafish lines.

Evaluation of the potassium adsorption capacity of a potassium adsorption filter during rapid blood transfusion.

The concentration of extracellular potassium in red blood cell concentrates (RCCs) increases during storage, leading to risk of hyperkalemia. A potassium adsorption filter (PAF) can eliminate the potassium at normal blood transfusion. This study aimed to investigate the potassium adsorption capacity of a PAF during rapid blood transfusion. We tested several different potassium concentrations under a rapid transfusion condition using a pressure bag. The adsorption rates of the 70-mEq/l model were 76·8%. The PAF showed good potassium adsorption capacity, suggesting that this filter may provide a convenient method to prevent hyperkalemia during rapid blood transfusion.

Corrective osteotomy for volar instability of the distal radioulnar Joint associated with radial shaft malunion.

Volar instability of the distal radioulnar joint is an uncommon wrist disorder. We report three cases of recurrent volar instability of the distal radioulnar joint secondary to fracture of the radial shaft. In all cases, X-rays showed a volar apex deformity of the radial shaft. Opening wedge osteotomy and iliac bone grafting was performed on the distal diaphysis of the radius instead of on the radial shaft, in order to adjust the distal radioulnar joint more easily. Pre-operative dislocations and painful clunks disappeared in all three patients. However, slight instability of the distal radioulnar joint remained in all cases. Osteoarthritis of the distal radioulnar joint was noted in one patient 31 months after the operation. All of the patients were satisfied with the results and did not desire further operations.

Dynamic gadolinium-enhanced MR imaging of pituitary adenomas: usefulness of sequential sagittal and coronal plane images.

Dynamic magnetic resonance (MR) imaging for pituitary adenomas is usually performed in a coronal direction; however, small lesions between slices, or lesions located at the anterior or posterior aspect of the pituitary gland might be overlooked on MR images in only the coronal direction. The purpose of our study was to evaluate whether consecutive dynamic MR images in the coronal and sagittal planes improve detection of pituitary adenomas. Eighteen patients with pituitary microadenomas and nine with healthy pituitary glands were included in this study. MR images were performed with 1.5 T superconductive units and commercially-available head coils. After a 5 ml gadolinium contrast injection, eight serial dynamic sagittal images were obtained. Within 3 or 6 min, this was followed by a 10-15 ml gadolinium injection and acquisition of eight serial dynamic coronal images. Dynamic MR images and conventional noncontrast- and contrast-enhanced sagittal and coronal T1-weighted images were evaluated independently in a blind fashion by two neuroradiologists regarding the depiction of pituitary microadenomas. The sensitivities of dynamic enhanced MR imaging in the detection of microadenomas were 61.1% in sagittal direction, 72.2% in coronal direction respectively, and were superior to those of conventional noncontrast- and contrast-enhanced T1-weighted imaging (22.2-50%). The sensitivity of a combination of sagittal and coronal dynamic enhanced MR imaging for the detection of microadenomas was 88.9% and was superior to those of conventional noncontrast- and contrast-enhanced T1-weighted imaging combining sagittal and coronal directions (61.1%, 61.1%) (P<0.05, P<0.05, respectively). The specificity and accuracy of dynamic enhanced MR imaging with combination of sagittal and coronal images was 88.9% respectively. Dynamic gadolinium-enhanced MR imaging, especially using both sagittal and coronal planes, was concluded to be useful for the detection of pituitary microadenomas.

Ideographic characters call for extra processing to correspond with phonemes.

Cortical areas used in the copying of Japanese ideographic characters and syllabic characters were studied using functional magnetic resonance imaging in healthy volunteers. Complexity of characters was controlled to illustrate differences resulting from character to sound conversion differences between the ideographic and syllabic characters. Statistical comparisons indicated extensive activation in the fusiform gyrus, posterior portions around the intraparietal sulcus and in the conjunction area of BA 6, 9 and 44 (which is assumed to be Exner's area) during the copying of ideographic characters. These findings suggested that indirectness between ideographic characters and their pronunciation demands extra processing such as semantic mediation and intensive grapheme processing in comparison with syllabic characters.

Tumour enhancement with newly developed Mn-metalloporphyrin (HOP-9P) in magnetic resonance imaging of mice.

The purpose of the study is to evaluate the tumour enhancing characteristics and biodistribution of a newly developed metalloporphyrin derivative, HOP-9P (13, 17-bis (1-carboxypropionyl) carbamoylethyl-3, 8-bis (1-phenylpropyloxyethyl)-2,7,12,18-tetra- methyl-porphynato manganese (III)). Seven mice bearing SCC VII tumours were imaged using T1-weighted conventional spin echo magnetic resonance images before and 5 min, 2 h and 24 h after intravenous injection of 0.1 mmol/kg of HOP-9P. For the acquired images, signal intensities of the tumour, muscle and oil-phantom were measured. Then, tumor/oil and tumor/muscle signal intensity ratios were calculated. Nineteen mice were sacrificed before or after the administration of HOP-9P (at 5 min, 2 h and 24 h), and the biodistribution of manganese in the tumour, muscle, liver, blood and kidneys was measured using optical emission spectrometers and was expressed as micrograms of manganese per gram of tissue. The tumour/muscle signal intensity ratio at 24 h (3.18 +/- 0.34) was significantly higher than precontrast ratio (1.77 +/- 0.20) (P < 0.05). The biodistribution assessment of manganese demonstrated that HOP-9P gradually and consistently accumulated in the tumour to reach the highest concentration at 24 h (3.49 +/- 1.22 micro gMn/g). It is concluded that HOP-9P is a potential tumour-specific MR contrast agent.

Assessment of gadolinium-enhanced time-resolved three-dimensional MR angiography for evaluating renal artery stenosis.

OBJECTIVE: The purpose of this study was to assess the image quality of gadolinium-enhanced time-resolved three-dimensional (3D) MR angiography and to evaluate its accuracy in revealing renal artery stenosis. SUBJECTS AND METHODS: Thirty-nine patients underwent MR angiography using an ultrafast 3D Fourier transform spoiled gradient-recalled acquisition in the steady state (TR/TE range, 2.6/0.7--0.8). Five seconds after administration of 15--20 mL gadodiamide hydrate, four or five consecutive data sets with imaging times of 7.0--7.6 sec were acquired during a single breath-hold. A timing examination was not performed. Image quality was assessed using quantitative analysis (signal-to-noise, contrast-to-noise, and venous-to-arterial enhancement ratios) and qualitative analysis (presence of venous overlap, presence of artifacts, and degree of renal arterial enhancement). MR angiography depiction of the renal artery stenosis was evaluated using conventional angiography as the standard of reference. RESULTS: On the best arterial phase, average aortic signal-to-noise ratio (+/-SD) was 74.5 +/- 24.4, aorta-to--inferior vena cava contrast-to-noise ratio was 70.8 +/- 23.4, and inferior vena cava--to-aorta venous-to-arterial enhancement ratio was 0.03 +/- 0.04. No venous overlap was seen in 38 of 39 patients. Substantial enhancement of renal arteries was seen in all patients without any noticeable artifacts. MR angiography correctly depicted the degree of stenosis in 44 of 47 normal arteries, 13 of 16 mildly stenotic arteries, five of five moderately stenotic arteries, three of four severely stenotic arteries, and one of one occluded artery. Sensitivity and specificity for revealing greater than 50% stenosis was 100%. CONCLUSION: Time-resolved 3D MR angiography can provide high-quality arteriograms. Its performance in revealing renal artery stenosis is comparable with that of conventional angiography.

Which morphologies of synovial folds result from degeneration and/or aging of the radiohumeral joint: an anatomic study with cadavers and embryos.

The synovial folds of the radiohumeral joints in cadaveric elbows from 179 elderly subjects and 40 embryos were investigated macroscopically and histologically to determine any morphologic changes caused by aging or degeneration. The anterior and posterior folds found in the elderly population shared characteristics of folds seen in embryos, with some modifications, and were thought to originate from the primitive septum. Proportionally, the length, width, and thickness of these folds were consistent between adults and embryos. However, the embryonic folds showed a homogenous morphology. In contrast, in the adult the anterior fold was characterized by a shorter and narrower villous pattern, and the posterior fold tended to be wider. Lateral extension of the anterior or posterior folds was also observed. Moreover, the lateral fold, never seen in embryos, was present and characterized by a hard plicate pattern in the adult. These derived or specific morphologies in adults probably result from alterations in the movement of the radial head caused by aging.

The vascular anatomy of the developing zebrafish: an atlas of embryonic and early larval development.

We have used confocal microangiography to examine and describe the vascular anatomy of the developing zebrafish, Danio rerio. This method and the profound optical clarity of zebrafish embryos make it possible to view the entire developing vasculature with unprecedented resolution. A staged series of three-dimensional images of the vascular system were collected beginning shortly after the onset of circulation at 1 day postfertilization through early- to midlarval stages at approximately 7 days postfertilization. Blood vessels in every region of the animal were imaged at each stage, and detailed "wiring patterns" were derived describing the interconnections between every major vessel. We present an overview of these data here in this paper and in an accompanying Web site "The interactive atlas of zebrafish vascular anatomy" online at (http://eclipse.nichd.nih.gov/nichd/lmg/redirect.html). We find a highly dynamic but also highly stereotypic pattern of vascular connections, with different sets of primitive embryonic vessels severing connections and rewiring in new configurations according to a reproducible plan. We also find that despite variation in the details of the vascular anatomy, the basic vascular plan of the developing zebrafish shows strong similarity to that of other vertebrates. This atlas will provide an invaluable foundation for future genetic and experimental studies of vascular development in the zebrafish.

Regional specificity in degenerative changes in finger joints: an anatomical study using cadavers of the elderly.

Rehabilitation therapists and orthopedists regard degenerative changes (DCs) of the articular cartilage or osteoarthritis in the finger joint as the major factors that interfere with normal grasp and pinch. However, previous research has concentrated on DCs in the carpometacarpal joint (CM) of the thumb. The present study revealed some general tendencies concerning the occurrence of DCs: (1) distal joint dominance between joints in a thumb or finger; (2) distal surface dominance in a joint; (3) radioulnar dominance in a joint; and (4) fifth-finger specificity (i.e., high incidences of both DCs and osteophytes). Moreover, according to our evaluation of multiple and solitary occurrences (i.e., how many segments carried DCs) we consider that DCs in the radioulnar segments advance not only to the complementary surface (i.e., mirror-image lesion formation) but also to other adjacent segments in the early stage before progression to cartilage defects occurs in the primarily affected segment. In addition, osteophytes were not associated with the cartilage defect in the adjacent segment, but, rather, occurred at opposite or distant segments in the joint. These results are discussed in relation to lateral pinch and precision grasping, with special reference to the hypothetical stress on these movements in the finger joint.

Interferon-gamma increases IL-12 mRNA expression and attentuates allergic late-onset airway responses in the Brown Norway rat.

Interferon gamma is a T-helper cell (Th)-1-type cytokine that has been suggested to inhibit the development of an atopic Th2-type profile of cytokine expression. The aim of this study was to investigated the effect of exogenous rat interferon gamma on antigen-induced airway responses, and on Th1 and Th2-type cytokine messenger ribonucleic acid (mRNA) expression in the Brown Norway rat. Rats were actively sensitized to ovalbumin and 14 days later underwent an aerosolized ovalbumin challenge. Animals were intratracheally administered either interferon gamma (3,000 U) or control solvent 30 min prior to, and 2 and 4 h following, antigen challenge. Lung resistance was monitored over an 8-h time period. Using in situ hybridization and immunocytochemistry, the levels of Th1- (interleukin-12) and Th2-type (interleukin4 and -5) cytokine mRNA, and major basic protein expression in the bronchoalveolar lavage fluid of these rats 8 h after ovalbumin challenge were also determined. Administration of interferon gamma attenuated the development of the late-onset airways response in ovalbumin-sensitized antigen-challenged rats (p<0,05). The expression of interleukin-4 and -5 mRNA in the bronchoalveolar lavage fluid of interferon gamma treated rats was significantly attenuated compared to ovalbumin-challenged saline-treated controls (p<0.001). This was accompanied by a significant increase in the expression of interleukin-12 mRNA, and a reduction in eosinophil numbers. Intratracheal administration of interferon gamma modulates the allergic late-onset airways response in rats, and this is associated with a reduction in the expression of T-helper cell 2-type cytokines and an increase in interleukin-12 messenger ribonucleic acid expression within the airways. The present results support a role for interferon gamma in the pathophysiology of acute allergic airway responses, possibly by virtue of its ability to modulate T-helper cell 1- 2-type cytokine expression within the lungs.

Dynamic MR dacryocystography: a new method for evaluating nasolacrimal duct obstructions.

OBJECTIVE: The purpose of this study was to evaluate the clinical performance of newly implemented dynamic MR dacryocystography. CONCLUSION: Dynamic MR dacryocystography, which requires neither ionizing radiation nor chemical contrast media with high viscosity, may be a useful tool for depicting nasolacrimal obstructions.

Software-triggered contrast-enhanced three-dimensional MR angiography of the intracranial arteries.

OBJECTIVE: We investigated the effectiveness of software-triggered contrast-enhanced three-dimensional (3D) MR angiography in evaluating intracranial arteries. SUBJECTS AND METHODS: We studied 38 patients with suspected brain lesions. Imaging was performed using a 1.5-T superconducting MR system with a commercially available head coil. To monitor signal intensity changes we used software to place a tracker volume at the basilar artery or the internal carotid artery. A 20-ml bolus of gadodiamide hydrate was administered through the antecubital vein at a rate of 2-4 ml/sec, followed by a saline flush. Three-dimensional MR angiography using a spoiled gradient-echo sequence with centric K-space ordering was triggered by the arrival of the contrast bolus in the tracker volume. Imaging times ranged from 12 to 20 sec. We used MR images to assess the effectiveness of contrast-enhanced 3D MR angiography in revealing intracranial arteries with minimal venous overlap. RESULTS: The software triggered imaging on the arrival of the contrast bolus in 81.6% of examinations. In 77.6% of examinations, the resulting MR angiograms revealed intracranial arteries with minimal venous overlap. CONCLUSION: Software-triggered contrast-enhanced 3D MR angiography with centric K-space ordering is a promising technique for viewing intracranial arteries.

An impairment of barrier size and charge selectivity of glomerular basement membrane in streptozotocin-induced diabetes and prevention by pharmacological therapy.

We examined barrier size (pore size) and charge selectivity (anionic sites) of the glomerular basement membrane (GBM) in experimental diabetes. For estimation of the pore size we employed a new method, tissue negative staining. In diabetic rats, enlarged pores and decreased numbers of anionic sites of GBM were observed. Both insulin treatment and angiotensin-converting enzyme inhibitors prevented these changes. The renoprotective effect of the two drugs is discussed in the article.

MRA of intracranial aneurysm models: a comparison of contrast-enhanced three-dimensional MRA with time-of-flight MRA.

PURPOSE: The purpose of our study was to compare contrast-enhanced 3D MR angiography (MRA) using ultrafast spoiled gradient-recalled acquisition in the steady state (SPGR) with 2D and 3D time-of-flight (TOF) MRA for visualization of intracranial lateral saccular aneurysm models. METHOD: We used lateral saccular aneurysm models with a height of 10 mm and neck sizes of 2.5 and 10 mm. Imaging was performed using a 1.5 T MR system with a head coil. Contrast-enhanced 3D MR angiograms were obtained using 3D ultrafast SPGR sequence with and without the MR Smartprep technique. Two-dimensional and 3D TOF MR angiograms were also obtained. RESULTS: Contrast-enhanced multiphase 3D MR angiograms taken every 5 s after injection of contrast medium proved superior to the other MRA techniques for delineating lateral saccular aneurysm models. Contrast-enhanced 3D MRA images taken with inadequate delay after MR Smartprep trigger showed poor visualization of the aneurysm model. CONCLUSION: Use of contrast-enhanced multiphase 3D MRA with ultrafast SPGR with shorter TR and TE resulted in clear images of the lateral saccular aneurysm model.

Morphogenesis of the primary arterial trunks of the forelimb in the rat embryos: the trunks originate from the lateral surface of the dorsal aorta independently of the intersegmental arteries.

It has been believed that the primary arterial trunk of the mammalian forelimb is derived from the 7th intersegmental artery. Here we examined the early morphogenesis of the arteries and nerves in the forelimb region by adopting a method that combined intravascular dye-injection with nerve staining to whole mounted rat embryos. The study was carried out on greater numbers of specimens at smaller intervals of embryonic stages and from earlier stages than those in previous reports. We report that: (1) The multiple primary arterial trunks in the forelimb region (primary subclavians) originate directly from the lateral surface of the dorsal aorta independently of the intersegmental arteries, previous to the formation of limb buds. (2) The tips of the 8th (and the 9th) primary subclavians that originate from the aorta near the origin of the 8th (or the 9th) intersegmental artery bend cranially and/or caudally. With the formation of limb bud, they extend to form the longitudinal trunks in the presumptive axillary region. The primary arteries in the free arm region branch off from this longitudinal trunk, and one of them develops into the axial artery. (3) The origins of the primary subclavians shift their positions on the surface of the dorsal aorta and approach the origins of the neighboring intersegmental arteries to join them, and then replace the latter. Consequently, the primary subclavians appear to be "the lateral branches of the in tersegmental arteries." (4) The 8th primary subclavian is dominant at first, but is replaced by the 7th primary subclavian, which develops into the definitive subclavian artery. (5) With the brachial nerve plexus formation, the axillary arterial plexus derived from the longitudinal trunk develops to form two stems of the axillary artery.

Initial ultrastructural changes in pore size and anionic sites of the glomerular basement membrane in streptozotocin-induced diabetic rats and their prevention by insulin treatment.

BACKGROUND/AIM: The present study was conducted to elucidate the mechanism(s) of the development of early diabetic nephropathy, examining ultrastructural changes employing electron microscopy, especially changes in pore size of the glomerular basement membrane (GBM) of streptozotocin (STZ)-induced diabetics rats. METHODS: Urinary albumin excretion rate (UAE), pore size of the lamina densa of the GBM visualized directly by the tissue negative staining method, and number of anionic sites (AS) in the corresponding portion of the lamina rara externa were determined for 6 weeks in diabetic rats without and with insulin treatment. RESULTS: The UAE of the diabetic rats increased with time and was significantly greater than that of the nondiabetic control rats after 4 weeks (p < 0.01), while insulin treatment suppressed the increased UAE of diabetic rats. The median values in both short diameter and long dimension of the pores in the diabetic group were markedly increased at the 2nd week as compared with those in the nondiabetic control rats, whereas no significant change was found in the pore size of the diabetic rats with insulin treatment. Moreover, the number of AS in the GBM of the diabetic rats was significantly (p < 0.001) decreased from the 2nd week onward. Insulin treatment also prevented a decrease in AS number in diabetic rats. CONCLUSIONS: It is suggested from these results that an impairment of barrier size selectivity occurs at a very early stage of STZ-induced diabetes in rats, which may enhance the abnormality of the charge-selective properties of the GBM. In addition, insulin treatment may protect this barrier system through normalizing blood glucose control in STZ-diabetic rats.

Tagged MR imaging of intracranial aneurysm models.

We used tagged MR imaging to investigate the flow in two lateral and two terminal saccular intracranial aneurysm models of different neck sizes. Imaging was performed with a 1.5-T superconducting MR system using 2D fast spatial modulation of magnetization (SPAMM) sequences with an intersegmental delay of 25 milliseconds. The flow in the saccular aneurysm models varied with the shape and size of the neck: flow was faster in wider-necked aneurysms than in those with narrower necks.

[Kinematic MRI using short TR single shot fast spin echo (SSFSE) in evaluating swallowing].

PURPOSE: To determine the utility of short TR single shot fast spin echo (SSFSE) MR imaging for evaluating swallowing. MATERIALS AND METHODS: Five healthy volunteers underwent kinematic MR imaging of swallowing with a 1.5T MR scanner using the short TR (300 ms) SSFSE sequence. Twenty phases of sagittal sections were acquired within 6 sec, where the temporal resolution was 300 ms. For oral contrast medium, we used prune yogurt juice with Fe added. RESULTS: The image contrast of short TR SSFSE was found to be somewhere like that of T1-weighted images. In all cases, both the buccal and pharyngeal stages of swallowing were successfully depicted. The Fe-added prune yogurt juice performed as a positive contrast medium and helped determine anatomical structures in the buccal stage. CONCLUSION: Short TR (300 ms) SSFSE was useful in evaluating swallowing. The combined use of Fe-added prune yogurt juice was helpful in enhancing the surface of the oropharynx.

[MR imaging of the gastrointestinal tract with half-Fourier single-shot fast spin echo (SSFSE)].

UNLABELLED: Our objective was to implement a non-invasive magnetic resonance imaging (MRI) technique combined with concentrated milk ingestion for depicting the gastrointestinal (GI) tract and detecting gastrointestinal motility and transit. The half-Fourier SSFSE (single-shot fast spin echo) sequence was optimized on the basis of a phantom study. In order to determine the feasibility of milk ingestion as a substitute for contrast medium, ten human volunteers were examined with SSFSE after two types of liquid ingestion (i.e., milk and water). The snapshot images provided subsecond data acquisition for each coronal plane, allowing visualization of peristalsis in the gastrointestinal tract in an almost real-time fashion, without motion-related image degradation, as would normally be seen using conventional MRI. RESULTS: There was no significant difference between concentrated milk and water in terms of depiction of the upper gastrointestinal tract; however, 10 min and 30 min after ingestion, concentrated milk showed better delineation of the intestine than that observed after water ingestion (p < 0.01). CONCLUSION: MR gastrointestinal imaging is a non-invasive method that allows gastrointestinal depiction as well as analysis of motility and passage. Especially with concentrated milk ingestion, the distal intestines were well depicted with adequate contrast filling and distention.