Optical imaging of mouse articular cartilage using the glycosaminoglycans binding property of fluorescent-labeled octaarginine.

OBJECTIVE: The aim of the current study was to examine the cartilage-specific binding property of polyarginine peptides (R4, 8, 12, and 16) and specifically to test octaarginine peptides for the optical imaging of articular cartilage in experimentally induced arthritis in mice. METHODS: Four rhodamine-labeled polyarginine peptides each with a different-length arginine chain (R4, 8, 12, or 16) were injected into the knee joints of C57BL/6J mice (n=20). The joints were excised 1h later and the fluorescent signal intensity in cartilage cryosections was compared for the four peptides. To examine the substrate of R8 in cartilage, femoral condyles obtained from another set of mice were treated with chondroitinase ABC (Ch'ase ABC), keratanase or heparitinase then immersed in R8-rhodamine. Fluorescent signals were examined by fluorescent microscopy. Next, R8-rhodamine was injected into the right knee joints of three control and three collagen antibody-induced arthritis (CAIA) mice, and fluorescent intensity in normal and degenerative cartilage was semi-quantitatively analysed on the histological sections using image software. Finally, femoral condyles from normal mice (n=2) and CAIA mice (n=2) were immersed in R8-rhodamine and calcein, then imaged using optical projection tomography (OPT). RESULTS: Fluorescent signals were specifically detected in the cartilage pericellular matrix from the surface to the tide mark but were completely absent in the calcified layer or bone marrow. The number of arginine residues significantly influenced peptide accumulation in articular cartilage, with R8 accumulating the most. The fluorescent signal in the femoral condylar cartilage diminished when it was treated with Ch'ase ABC. R8 accumulation was significantly decreased in the degenerative cartilage of CAIA mice, and this was demonstrated both histologically and in three-dimensional (3D)-reconstruction image by OPT. CONCLUSION: R8 may be a useful new experimental probe for optical imaging of normal and arthritic articular cartilage.

An effective combination of carbohydrases that enables reduction of dietary protein in broilers: importance of hemicellulase.

After observing the effects of purified cellulase (C), hemicellulase (H), pectinase (P), and their combinations on the in vitro digestibility of a corn-soybean meal broiler diet, we examined the associations between pectin breakdown and the digestibilities of CP and DM by using free galacturonic acid (GA) as an index of pectin breakdown. There was no significant effect of the single enzymes except for H. However, the enzyme combinations H + P, C + H, and C + H + P significantly increased CP and DM digestibilities, whereas the combination of C + P was not effective. Because H has activities of both H and P, these enzymes were considered to be important in stimulating digestion. Furthermore, when the enzymes increased CP and DM digestibilities, GA concentration was significantly higher, and clear correlations between CP and DM digestibilities and GA concentration were observed, whereas correlations between the digestibilities and concentration of glucose or xylose + mannose as indices of cellulose and hemicellulose breakdown, respectively, were not significant. From these observations, we hypothesized that a mixture of enzymes could increase the protein digestibility of broiler feed. Thus, in the in vivo experiment, low-protein (19% CP) diets made mainly of corn and soybean meal with or without mixed enzymes were prepared and given to broiler chicks. The birds given the diet containing mixed enzymes showed significantly higher BW gain, with higher CP and DM digestibilities than the birds given the diet without the mixed enzymes. Moreover, the growth rate was same as that of the birds given the normal (21% CP) diet. The results indicate that the mixed enzyme preparation can effectively degrade indigestible cell constituents and thus enable the protein of the broiler feed to become more digestible. Furthermore, the results indicate the importance of H as a rate-limiting factor of cell wall breakdown.

Immunohistochemical localization of histamine N-methyltransferase in guinea pig tissues.

Histamine plays important roles in gastric acid secretion, inflammation, and allergic response. Histamine N-methyltransferase (HMT; EC 2.1.1.8) is crucial to the inactivation of histamine in tissues. In this study we investigated the immunohistochemical localization of this enzyme in guinea pig tissues using a rabbit polyclonal antibody against bovine HMT. The specificity of the antibody for guinea pig HMT was confirmed by Western blotting and the lack of any staining using antiserum preabsorbed with purified HMT. There was strong HMT-like immunoreactivity (HMT-LI) in the epithelial cells in the gastrointestinal tract, especially in the gastric body, duodenum, and jejunum. The columnar epithelium in the gallbladder was also strongly positive. Almost all the myenteric plexus from the stomach to the colon was stained whereas the submucous plexus was not. Other strongly immunoreactive cells included the ciliated cells in the trachea and the transitional epithelium of the bladder. Intermediately immunoreactive cells included islets of Langerhans, epidermal cells of the skin, alveolar cells in the lung, urinary tubules in the kidney, and epithelium of semiferous tubules. HMT-LI was present in specific structures in the guinea pig tissues. The widespread distribution of HMT-LI suggests that histamine has several roles in different tissues.

Ectopic osteogenesis with biphasic ceramics of hydroxyapatite and tricalcium phosphate in rabbits.

Porous calcium phosphate ceramics consisting of hydroxyapatite (HA) and tricalcium phosphate (TCP) with different HA to TCP ratio were implanted intramuscularly in rabbits for six months in order to carry out a comparative study on osteogenic activity of the ceramics. Bone formation was detected only in HT73 (HA to TCP ratio, 7-3) specimens. Other implants, HT28 (2-8) and HT010 (0-10), could not induce bone. After a six-month period of implantation, HT28 and HT010 implants showed obvious degradation of the implants changing their shape and size macro and microscopically. Microscopically, they showed aggregates of fine particles and appearance of multinucleated cells. However, HT73 implants was less degraded and could maintain their original structure macro and microscopically. This study showed that HT73 ceramics can induce bone in rabbit muscle tissue and it is considered that maintenance of porous structure, that is, degradation rate of the materials may be one of the affecting factors in ceramic-induced osteogenesis.

Pancreatic insulo-acinar portal systems in humans, rats, and some other mammals: scanning electron microscopy of vascular casts.

Scanning electron microscopy of vascular casts showed that in the mouse, rat, and guinea pig, the pancreatic endocrine islets were frequently interlobular in position and emitted insulo-venous efferent vessels directly draining into veins. In these animals, the intralobular islets, located within the exocrine lobules, issued insulo-acinar portal vessels continuous with the lobular capillaries in addition to the insulo-venous efferent vessels. In humans, monkeys, cows, pigs, dogs, cats, and rabbits, essentially all islets in the pancreas were intralobular in location and emitted the insulo-acinar portal vessels only. In man and animals examined, especially in the murine species, many lobules lacked an islet, therefore the insular control over the exocrine pancreas seemed to be effected in more or less restricted areas of lobules.

Blood vascular bed and pericapillary space in rat parathyroid glands.

The blood vascular bed and pericapillary space of the rat parathyroid gland were studied by scanning electron microscopy of vascular casts, freeze-cracked tissue blocks, and NaOH-treated tissue specimens. The findings were supplemented by transmission light and electron microscopy of sectioned tissue samples. The rat parathyroid gland contained a rich network of freely anastomosing capillaries. These capillaries were surrounded by marked pericapillary spaces that were demarcated by basal lamina of both capillaries and parenchymal cells. The pericapillary spaces contained many collagen fibrils and frequently issued some projections running deep into the sheets of parathyroid cells. The latter projections may be useful to supply the parenchymal cells located far from the capillaries. The collagen fibrils may regulate the flow of tissue fluid in the pericapillary space and convey parathyroid hormone, which is released at the apicolateral domain, into the capillaries.

Blood flow patterns in the rat pancreas: a simulative demonstration by injection replication and scanning electron microscopy.

Scanning electron microscopy of vascular casts prepared by arterial injections of intentionally reduced amounts of resin showed that in the rat pancreas, the casting medium fills blood capillaries in the endocrine islets more promptly than those in the exocrine lobules and secretory ducts. Furthermore, the exocrine lobules containing endocrine islets allowed a more rapid resin flow through the insulo-acinar portal route than those lobules lacking an islet. The capillaries of secretory ducts were the last portions to be filled with resin. Since the resin used in this study was as viscous as blood and injected under a physiological pressure, the microcirculatory patterns demonstrated by the present method reflect the physiological flow pattern of blood in the pancreas.

Distribution of adrenomedullin-containing perivascular nerves in the rat mesenteric artery.

Distribution of adrenomedullin (AM)-containing perivascular nerve fibers was studied in rat mesenteric arteries. Many fibers containing AM-like immunoreactivity (LI) were observed in the adventitia. AM-LI fibers were abolished by cold storage denervation or capsaicin but not 6-hydroxydopamine. Double immunostainings showed colocalization of AM-LI with calcitonin gene-related peptide (CGRP)-LI. The dorsal root ganglia had many AM-positive cells and AM mRNA detected by RT-PCR. Electron microscopy study revealed high proportions of immunogold labeling for AM and colocalization of both AM-LI and CGRP-LI in unmyelinated nerve axons. These results suggest that AM-containing perivascular nerves are distributed in the rat mesenteric artery.

A novel serpin-like protein, B-43, exists in both neurons and astrocytes: an immunohistochemical study in the parietal region of the bovine brain.

The presence of a novel member of serine proteinase inhibitor, B-43, was immunohistochemically indicated in both neurons and astrocytes in the parietal region of the bovine brain. B-43-like immunoreactivity was detected in pyramidal cells in the cortex and GFAP-positive astroglial cells in the white matter. The processes of B-43 may play a cooperative role with glia-derived nexin/protease nexin-1 and alpha 1-antichymotrypsin in the brain.

Basic imaging properties of a computed radiographic system with photostimulable phosphors.

We measured the characteristic curve, modulation transfer function (MTF), and the Wiener spectrum of a commercially available computed radiographic (CR) system with photostimulable phosphor plate (imaging plate, IP). The characteristic curve (system response) obtained by an inverse-square x-ray sensitometry showed a wide dynamic range (order of 10(3) in maximum). The slit technique was employed to determine the MTF's, such as IP MTF, presampling MTF including the unsharpness of the detector (IP) and the blurring effect of the sampling aperture, and laser-printer MTF. It was found that the MTF of the standard type of IP was comparable to that of medium-speed screen/film systems. The noticeable degradation of resolution in our CR system, however, occurred at the stage of image data sampling: the presampling MTF was inferior to the IP MTF due to the effect of the scattering and resultant spreading of the incidence laser beam and the emitted luminescence. The noise was characterized by means of digital Wiener spectrum using uniformly exposed noise data. Exposure ranges could be separated into different sections depending upon the noise sources, such as quantum mottle at low exposure and system structure noise at high exposure.

Plasma and testicular testosterone in experimental diabetic rats.

Testicular and plasma testosterone levels were found to be decreased markedly in streptozotocin diabetic rats compared with those of controls. Treatment with 6 units NPH insulin daily for one week almost normalized plasma testosterone levels parallel to the increase in body and liver weights in diabetic rats, while testosterone levels in testicles were not significantly changed. Plasma prolactin and LH levels were unchanged among control, diabetic and diabetic insulin-treated rats. Thus, testosterone reduction in the testis might play a role in diabetic impotence.

Effect of prostacyclin infusion on active and inactive renin release in the isolated perfused kidney.

The effect of prostacyclin infusion into the renal artery of the isolated perfused hog kidney on the release of active and inactive renin was investigated. Infusion of prostacyclin at a rate of 0.1 microgram/min resulted in a significant increase (p less than 0.01) in active renin and a significant fall (p less than 0.01) in inactive renin. Prostacyclin also increased urinary kallikrein excretion (p less than 0.05). The results indicate that the kidney secretes not only active renin but also inactive renin, and suggest that prostacyclin stimulates the conversion of inactive renin to the active form through the activation of the renal kallikrein system.

A simple method for determining the modulation transfer function in digital radiography.

The authors developed a simple method for determining the presampling modulation transfer function (MTF). which includes the unsharpness of the detector and the effect of the sampling aperture, in digital radiographic (DR) systems. With this method, the presampling MTF is determined by the Fourier transform of a ;finely sampled' line spread function (LSF) obtained with a slightly angulated slit in a single exposure. Since the effective sampling distance becomes much smaller than the original sampling distance of the DR system, the effect of aliasing on the MTF calculations can be eliminated. The authors applied this method to the measurement of the presampling MTF of a compound radiographic system and examined the directional dependence, the effect of exponential extrapolation, and the effect of different sampling distances. It is shown that the technique of multiple slit exposure and exponential extrapolation of the LSF tail, which has been commonly used in analog seven-film systems, can be employed in DR systems. The authors determined the glare fraction in order to estimate the component of low-frequency drop mainly due to ;glare'

Obstructive hydrocephalus due to a giant aneurysm of the internal carotid bifurcation.

A rare case of obstructive hydrocephalus associated with a giant aneurysm of the internal carotid artery is presented. The giant aneurysm, which was located in the prepontine cistern, was initially thought to be of basilar artery origin. Cerebral computed angiotomography had the most diagnostic value in disclosing the partially thrombosed giant aneurysm and its relationship with the surrounding arteries. After undergoing a ventriculoperitoneal shunt operation, the patient's clinical symptoms markedly improved.

Resorption of the postero-superior corner of the lateral part of the mandibular condyle correlates with progressive TMJ internal derangement.

The purpose of this study was to test the hypothesis that resorption of postero-superior corner of lateral part of the mandibular condyle (RLC) occurs with the advancement of TMJ internal derangement (ID). One hundred and sixty-seven TMJs in 122 patients were studied. RLC was assessed on plane radiographs of frontal oblique projection. The correlation between the prevalence of RLC and radiological stage of ID was analysed and clinical correlations to the radiological stage of ID and the prevalence of RLC were discussed. The higher prevalence of RLC in joints with advanced ID was statistically significant (Mann-Whitney U-test, P< 0.05). There was no correlation between the prevalence of RLC and neither OA on the articular surface of the condyle nor clinical symptoms/signs. The result of this study suggests that RLC appears to occur in joints with advanced ID as a result of remodelling changes secondary to ID.

The interosseous and lumbrical muscles in the human hand, with special reference to the insertions of the interosseous muscles.

The interosseous and lumbrical muscles in twenty-five hands of Japanese adult cadavers were dissected. The palmar and dorsal interosseous muscles continued, with few exceptions, into the wing tendons. The dorsal interosseous muscles gave off tendons which pierced the transverse laminae or passed deep to the transverse laminae, and attached to the bases of the proximal phalanges. The palmar interosseous muscles seldom had such attachments. The palmar and dorsal interosseous muscles sometimes gave off additional tendons which passed superficial to the transverse laminae and attached to the bases of the proximal phalanges. These latter attachments were typical in the contrahentes muscles. Thus, the present findings suggest that the human dorsal interosseous muscles are composite muscles derived from the dorsal abductor, flexor brevis and contrahens muscles, and that the human palmar interosseous muscles are composite muscles derived from the flexor brevis and contrahens muscles. The lumbrical muscles rarely gave off accessory slips with atavistic attachments to the proximal phalanges.

Homology of the adductor pollicis and contrahentes muscles: a study of monkey hands.

The deep palmar muscles in monkey hands were studied. The contrahentes muscles mainly arose from the capitate bone, descended palmar to the deep palmar branch of the ulnar nerve and the palmar metacarpophalangeal nerves, and attached to the proximal phalanges or wing tendons of the second, fourth and fifth fingers. In relation to the deep palmar branch of the ulnar nerve and the palmar metacarpophalangeal nerves, the contrahentes muscles are homologous with the adductor pollicis and flexor indicis radialis muscles. The contrahentes muscles occasionally gave off some accessory slips which blended with the interosseous muscles. These findings suggest that the human adductor pollicis muscle is a well-developed remnant of a contrahens muscle, and that the human interosseous muscles contain some remnant of the contrahentes muscle. In fact, a well-developed remnant of a contrahens muscle was found in the fourth finger of a human hand. It is further considered that the human adductor pollicis muscle contains an element of the interosseous muscle of the thumb.

Typology of abdominal arteries, with special reference to inferior phrenic arteries and their esophageal branches.

Origins and distribution of the human inferior phrenic arteries were studied by dissecting 68 Japanese adult cadavers. The inferior phrenic arteries were usually observed as paired (left and right) vessels. Their origins were summarized as follows: a) the aorta itself (85/138 cases, 61.6%), b) the ventro-visceral arteries (celiaco-mesenteric system of the aorta) including the celiac trunk (39/138 cases, 28.2%) and the left gastric artery (4/138 cases, 2.9%), and c) the latero-visceral arteries (adreno-renal system of the aorta) including the middle adrenal artery (4/138 cases, 2.9%) and the renal artery (6/138 cases, 4.3%). The left and right arteries occasionally originated in common trunk from the aorta, celiaco-mesenteric system or adreno-renal system (22/138 cases, 15.9%). A typological diagram explaining these variations is given. The inferior phrenic arteries, especially the left ones, sometimes issued visceral or esophageal branches. This fact indicates that the inferior phrenic arteries are homologous with the celiac trunk and mesenteric arteries. It is further discussed that the celiac trunk and mesenteric arteries are originally paired vessels, through introduction of our previous typological diagram of the abdominal arteries.

Perineuronal sulfated proteoglycans, cell surface glycoproteins and dark neurons in the cingulate cortex of newborn and adult rats.

Many neurons in the adult rat cingulate cortex possess perineuronal sulfated proteoglycans detectable with cationic iron colloid and aldehyde fuchsin, or cell surface glycoproteins reactive to lectin Vicia villosa or soybean agglutinin. The perineuronal sulfated proteoglycans develop three to four weeks after birth. The cell surface glycoproteins develop at earlier stage or two to three weeks after birth. Dark or active neurons begin to appear three to four weeks after birth. These findings indicate that the brain matures after birth or during weaning period.

Perineuronal sulfated proteoglycans in the adult rat brain: histochemical and electron microscopic studies.

Neurons of cerebellar nuclei in the rat brain had a marked surface coat which was stained with cationic iron colloid or aldehyde fuchsin. Neurons with a similar surface coat were also noted in the retrosplenial cortex. The surface coat was stained doubly with cationic iron colloid and aldehyde fuchsin. Digestion with hyaluronidase eliminated the stainability of the surface coat to both agents. Combined digestion with chondroitinase ABC, heparitinase and keratanase eliminated the cationic iron colloid staining but did not interfere with the aldehyde fuchsin staining. Electron microscopy of ultrathin sections revealed that the iron particles were deposited in the perineuronal tissue spaces. These findings indicate that the surface coat consists of sulfated proteoglycans which occupy, as the extracellular matrix, the perineuronal tissue spaces. Many neurons in the retrosplenial cortex were labeled with lectin Vicia villosa agglutinin. Double staining revealed that these lectin-labeled neurons are usually reactive to cationic iron colloid. Few neurons in the cerebellar nuclei were labeled with lectin V. villosa agglutinin.