Histology and chronological magnetic resonance images of congenital spinal deformity: An experimental study in mice model.

BACKGROUND: The natural history of the congenital spinal deformity and its clinical magnitude vary widely in human species. However, we previously reported that the spinal deformities of congenital scoliosis mice did not progress throughout our observational period according to soft X-ray and MRI data. In this study, congenital vertebral and intervertebral malformations in mice were assessed via magnetic resonance (MR) and histological images. METHODS: Congenital spinal anomalies were chronologically assessed via soft X-ray and 7 T MR imaging. MR images were compared to the histological images to validate the findings around the malformations. RESULTS: Soft X-ray images showed the gross alignment of the spine and the contour of the malformed vertebrae, with the growth plate and cortical bone visible as higher density lines, but could not be used to distinguish the existence of intervertebral structures. In contrast, MR images could be used to distinguish each structure, including the cortical bone, growth plate, cartilaginous end plate, and nucleus pulposus, by combining the signal changes on T1-weighted imaging (T1WI) and T2-weighted imaging (T2WI). The intervertebral structure adjacent to the malformed vertebrae also exhibited various abnormalities, such as growth plate and cartilaginous end plate irregularities, nucleus pulposus defects, and bone marrow formation. In the chronological observation, the thickness and shape of the malformed structures on T1WI did not change. CONCLUSIONS: Spinal malformations in mice were chronologically observed via 7 T MRI and histology. MR images could be used to distinguish the histological structures of normal and malformed mouse spines. Malformed vertebrae were accompanied by adjacent intervertebral structures that corresponded to the fully segmented structures observed in human congenital scoliosis, but the intervertebral conditions varied. This study suggested the importance of MRI and histological examinations of human congenital scoliosis patients with patterns other than nonsegmenting patterns, which may be used to predict the prognosis of patients with spinal deformities associated with malformed vertebrae.

Two synergistic types of muscles were detected during forearm rotation exercise by T2 cumulative frequency curves using 0.2 T magnetic resonance imaging.

The purpose of this study was the detection and characterization of synergistic muscle activity. Using T2-map MRI, T2 values for 10 forearm muscles in 11 healthy adult volunteers were obtained in the resting state and after isotonic forearm supination and pronation exercises with the elbow extended. T2 was normalized by Z = (T2e-T2r)/SDr, where T2e was T2 after exercise, while T2r and SDr were the reference values of 34 ms and 3 ms, respectively. Using the cumulative frequency curves of Z values (CFZ), we detected 2 and 3 synergistic muscles for supination and pronation, respectively, and divided these into 2 types, one activated by exercise strength dependently, and the other, independent of exercise strength, activated by only a smaller fraction of the participants. We also detected co-contraction for the supination. Thus, CFZ is a useful visualization tool to detect and characterize not only synergistic muscle, but also co-contraction muscle.

Pulsative venous return from the branchial vessels to the heart of the bivalve Mytilus galloprovincialis supports the constant-volume mechanism.

In bivalves and gastropods, ventricle contraction causes a negative pressure in the auricles and increases venous return from the afferent oblique vein (AOV): the constant-volume (CV) mechanism. The flow in the AOV should be a pulsative flow synchronized with the ventricular contraction. The flow in the heart and adjacent vessels of Mytilus galloprovincialis were measured by magnetic resonance imaging to confirm this hypothesis. Under a regular heartbeat, pulsative flows in the AOV and branchial vessels (BVs) were almost completely synchronized with the flow in the aorta, while filling of the ventricle was in the opposite phase. Flows in the BVs were directed to the posterior direction, and a pair of BVs in the gill axes (the efferent BVs) were connected to the AOV. Based on the images of the whole pathway of the AOV in an oblique slice, we confirmed that haemolymph flow was evoked from the efferent BVs and flow into the ventricle via the auricle was completed in a single heartbeat. Therefore, the walls of the AOV and BVs could resist negative transmural pressure caused by the ventricular contraction. In conclusion, the auricle, the AOV and the BVs, including the gill filaments, act as a suction pump. The pulsative venous return is driven by the negative pressure of the AOV as in the CV mechanism, and the negative pressure in the efferent BVs could draw haemolymph from the sinus via the gill and the afferent BVs. Therefore, Mytilus can start and stop its heartbeat as necessary.

Central administered xenin induced Fos expression in nesfatin-1 neurons in rats.

Xenin is a 25-amino acid peptide identified in human gastric mucosa, which is widely expressed in peripheral and central tissues. It is known that the central or peripheral administration of xenin decreases food intake in rodents. Nesfatin-1/NUCB2 (nesfatin-1) has been identified as an anorexic neuropeptide, it is often found co-localized with many peptides in the central nervous system. After the intracerebroventricular administration of xenin on nesfain-1-like immunoreactivity (LI) neurons, we examined its effects on food intake and water intake in rats. As a result, Fos-LI neurons were observed in the organum vasculosum of the laminae terminalis (OVLT), the median preoptic nucleus (MnPO), the subfornical organ (SFO), the supraoptic nucleus (SON), the paraventricular nucleus (PVN), the arcuate nucleus (Arc), the lateral hypothalamic area (LHA), the central amygdaloid nucleus (CAN), the dorsal raphe nucleus (DR), the locus coeruleus (LC), the area postrema (AP) and the nucleus of the solitary tract (NTS). After the administration, the number of Fos-LI neurons was significantly increased in the LC and the OVLT, the MnPO, the SFO, the SON, the PVN, the Arc, the LHA, the CAN, the DR, the AP and the NTS, compared with the control group. After the administration of xenin, we conducted double immunohistochemistry for Fos and nesfatin-1, and found that the number of nesfatin-1-LI neurons expressing Fos were significantly increased in the SON, the PVN, the Arc, the LHA, the CAN, the DR, the AP and the NTS, compared with the control group. The pretreatment of nesfatin-1 antisense significantly attenuated this xenin-induced feeding suppression, while that of nesfatin-1 missense showed no improvement. These results indicate that central administered xenin may have anorexia effects associated with activated central nesfatin-1 neurons.

Foot extension and retraction in the clam Calyptogena okutanii without any Keber's valve: an inflatable fastener bag model.

In order to investigate the foot manipulation of a clam without a Keber's valve, Calyptogena okutanii was examined by light microscopy, magnetic resonance imaging and computed tomography. The foot chamber was divided into two compartments by a dense muscle fastener zone (FZ) comprising a pedal artery and sinuses in the mid-sagittal plane in between muscles running in the anterior-posterior oblique direction. The distal part of the foot chamber (inflatable fastener bag, IFB) had a loose superficial muscle layer. The proximal part of the foot chamber (visceral reservoir, VR) was covered by a dense superficial muscle layer. The outlet of the VR was connected with the hinge ligament duct, consisting of the hinge ligament, a pair of shells and the pericardium. Based on these anatomical structures, foot extension starts from contraction of muscles in the FZ, so that flow in the FZ is stopped. Then, the superficial muscles of the foot contract, and the pressure of the IFB increases so that the foot can extend. Foot retraction starts from the relaxation of muscles in the FZ so that the hemolymph returns to the VR. The hinge ligament duct allows a constant return flow from the foot chamber to the gills and the heart. The heart rate and the flow in the FZ, which decreased and increased during the foot extension and retraction, respectively, supported this model. In conclusion, the FZ of Calyptogena okutanii could be an alternative to Keber's valve in Anodonta, playing a similar role.

The kidney of the Nodularia freshwater mussel has a larger filtration-size and counter-current system with improved water excretion compared with the seawater mussel Mytilus.

Histological studies and magnetic resonance imaging were employed to analyze the kidney structure and function of the freshwater mussel, Nodularia douglasiae. The Nodularia kidney consists of proximal, intermediate and distal tubules. The epithelia of the renal tubules were composed of a single layer of cuboidal cells. The proximal and distal tubules run in opposite directions underneath the pericardial cavity. Molecular weight cut-off (MWCO) values for the kidney filtration were detected by MR tracer injections: gadolinium-diethylenetriaminepentaacetic acid (GdDTPA) at 0.55 kDa, an oligomer-based contrast agent (CH3-DTPA-Gd) at 2.2 kDa, as well as Gd-DTPA-polylysine at 10, 22, and 110 kDa. The T1w-MRI intensity and T1 relaxation rate (R1) of the pericardial cavity and renal tubules increased with tracers smaller than 10 kDa. The other tracers showed only minimal or no increase. Thus, we concluded that the MWCO of the kidney is 22 kDa, 50 times larger than that for the Mytilus living in seawater. Since the R1 values of the renal tubules were similar to those of the pericardial cavity, the kidney did not concentrate filtrated tracers. The slow decay of the magnetic resonance (MR) tracers from the renal tubules indicated a low filtration rate, suggesting that the counter-current system reabsorbs useful solutes without reabsorption of water. The higher MWCO may be beneficial to maintain the tubular oncotic pressure and allow excretion of excess water. In conclusion, a main renal function of the freshwater mussel is the excretion of water, opposite to that of the seawater mussel and vertebrates, which preserve water.

MRI-based kinematics of the menisci through full knee range of motion.

PURPOSE: The meniscal kinematics in the full knee range of motion (ROM) have not been demonstrated by MRI, because the narrow bore of the superconducting magnet prevents full knee motion. The purpose of this study was to the investigate meniscal kinematics associated with femorotibial kinematics using an open-structure MRI unit that allows kinematic analysis of the menisci in full knee ROM. METHODS: Non-weight-bearing MR images of the right knee of 10 subjects were acquired at six angles of knee flexion (0°, 30°, 60°, 90°, 120°, and full flexion) using a compact 0.2-T MRI system. The positions of the anterior and posterior horns of the medial and lateral menisci (MM/LM) and the medial and lateral femoral condyles (MFC/LFC) were measured at each angle of flexion. RESULTS: Significant posterior LFC movement was observed in all sets of adjacent flexion angles of 60°-90° or more, indicating medial pivot motion of the femur. Significant differences in LM position were observed between adjacent flexion angles of 60°-90° or more. The positional relationship between the posterior horn of MM and the MFC was statistically significant in all but 60° flexion. The positional relationship between LM and LFC was significant at flexion angles of ≤90° in the anterior horn and at 60°, 90°, and full flexion in the posterior horn. CONCLUSION: Motion patterns of the menisci were analogous to those of the femoral condyle. Medial pivot motion of the femur caused the greatest posterior movement of the LM. Meniscal kinematics followed the femorotibial kinematics. Comprehension of meniscal kinematics in full knee ROM is important for understanding of injury mechanisms, planning meniscus transplant, and making postoperative care program for meniscus surgery.

Microhemorrhage in a Rat Model of Neonatal Shaking Brain Injury: Correlation between MRI and Iron Histochemistry.

Previous studies have shown that neonatal shaking brain injury (SBI) causes transient microhemorrhages (MHs) in the gray matter of the cerebral cortex and hippocampus. Iron deposits and iron-uptake cells are observed surrounding MHs in this SBI model, suggesting local hypoxic-ischemic conditions. However, whether the shaken pups suffered systemic hypoxic-ischemic conditions has remained uncertain. Further, histopathological correlations of MHs on magnetic resonance imaging (MRI) are still unclear. The present study examined MHs after neonatal SBI using a combination of histochemical and susceptibility-weighted imaging (SWI) analyses. Systemic oxygen saturation analyses indicated no significant difference between shaken and non-shaken pups. MHs on postnatal day 4 (P4) pups showed decreased signal intensity on SWI. Iron histochemistry revealed that these hypointense areas almost completely comprised red blood cells (RBCs). MHs that appeared on P4 gradually disappeared by P7-12 on SWI. These resolved areas contained small numbers of RBCs, numerous iron-positive cells, and punctate regions with iron reaction products. Perivascular iron products were evident after P12. These changes progressed faster in the hippocampus than in cortical areas. These changes in MHs following neonatal SBI may provide new insights into microvascular pathologies and impacts on brain functions as adults.

Detection of muscle activity with forearm pronation exercise using T2-map MRI.

[Purpose] We aimed to detect muscle activity during a forearm pronation exercise using a 0.2 T MRI system. [Participants and Methods] We recruited healthy adult volunteers (7 males, 4 females). Transverse relaxation time (T2) values for 10 forearm muscles were obtained from transverse multiple-spin-echo MR images of one-third of the ulna, lengthwise from the olecranon, in the resting state and after isotonic forearm pronation exercise at three strength levels (5, 15, and 25% of the maximum voluntary contraction). Z values were calculated as (T2e - T2r)/SDr, where T2e, T2r and SDr were T2 after exercise, 34 ms, and 3 ms, respectively. A Z value of 2.56 was used as the threshold for defining muscle activation. [Results] T2 values increased significantly in the pronator teres muscle (agonist), while those in the supinator muscle (antagonist) showed no change. The sensitivity and specificity values obtained were high and low, respectively, for all of the three exercise strength levels employed. In some of the participants, activity was detected in the flexor carpi radialis, extensor carpi ulnaris, and extensor digitorum. [Conclusion] Using T2-map MRI, we detected activity in primary and secondary mover muscles. We also found individual variations in the use of forearm muscles during pronation.

Rat mandibular condyle and fossa grew separately then unified as a single joint at 20 days old, which was the weaning age.

Magnetic resonance imaging (MRI) was used to observe growth of the mandibular condyle, mandibular fossa, and articular disc as a single unit. Changes in each component's relative position and size were observed using 7-tesla MRI. Mandibular condyle chondrocytes' growth was evaluated with immunohistochemistry, using the expression of zinc transporter ZIP13. Three-dimensional T1-weighted (T1w) MRI was used to obtain images of the TMJ of Sprague Dawley rats at 4-78 days old (P4-78) with a voxel resolution of 65 µm. Two-dimensional T1w MR images were acquired after a subcutaneous injection of the contrast reagent gadolinium diethylenetriamine pentaacetic acid (Gd-DTPA). The T1w MR images showed that the mandibular condyle was located posterior to the mandibular fossa until P20; however, it then moved to a location underneath the mandibular fossa. In the Gd-DTPA enhanced images, the articular disc was identified as a region with lower signal intensity from P20. The number of ZIP13-positive chondrocytes at P6 was larger than the number at P24. In conclusion, the mandibular condyle with cartilage and disc grows on the posterior side of the mandibular fossa until P20, which was the weaning age. Then, the condyle fit into the mandibular fossa and completed the functional unit.

Size-selective filtration of the atrial wall estimated from the accumulation of tracers in the kidney of the mussel Mytilus galloprovincialis.

In order to determine the molecular weight cut-off (MWCO) for the atrial wall filtration into kidneys of the Mytilus galloprovincialis, we employed five magnetic resonance (MR) tracers: manganese chloride (Mn2+), gadolinium chloride (Gd3+), manganese-ethylenediaminetetraacetic acid (MnEDTA), gadolinium-diethylenetriamine pentaacetic acid (GdDTPA) and oligomer-based contrast agent (CH3-DTPA-Gd). After injection of the MR tracers (1 or 2 mmol l-1×0.1 ml) into the visceral mass, T1-weighted MR imaging (T1w-MRI) and the longitudinal relaxation rates (1/T1=R1) were measured at 20°C. The MR tracers were distributed uniformly in the visceral mass within 1 h after injection. The T1w-MRI intensity and R1 of the kidney (R1K) were increased by Mn2+ and MnEDTA, with urine concentrations estimated at 210 and 65 µmol l-1, respectively. The rest of the tracers showed only minimal or no increase. When the mussels were additionally incubated in seawater with 10 µmol l-1 MnCl2, R1K was increased in the GdDTPA group, but not in the GdCl3 group. Therefore, Gd3+ might have inhibited renal accumulation of Mn2+ and Gd3+ Incubation in seawater with 10 µmol l-1 MnEDTA showed no increase in the R1K, but additional incubation with 10 µmol l-1 MnCl2 caused an increase in R1K It is suggested that injected MnEDTA was filtrated as MnEDTA per se, and not likely separated into free Mn2+ Thus, we concluded that the MWCO of the atrial wall of the M. galloprovincialis is around 0.5 kDa, which is almost 1/100 of that for vertebrate animals, and suggests a reduction in efforts to reabsorb metabolites and osmolytes from the urine.

T2 Distribution in the Forearm Muscles and the T2 Threshold for Defining Activated Muscle.

PURPOSE: In order to detect exercised muscles by the increase in T2, we have defined a Gaussian T2 distribution and reference values (T2r and SDr) in resting state muscles, and a threshold for detecting exercised muscles. METHODS: The subjects were healthy adult volunteers (14 males and 12 females). Multiple-spin-echo (MSE) MR images were obtained with 10 TE values from 10 to 100 ms using a 0.2T MRI system. T2 values for 10 forearm muscles were obtained in the resting state and after isometric wrist flexion exercise with 5%, 15%, and 25% of the maximum voluntary contraction (MVC). Z values were obtained by (T2e - T2r)/SDr, where T2e was T2 after exercise. Based on sample size calculations, three thresholds (ZT = 1.00, 2.56, and 3.07) were applied to agonist and antagonist muscles. RESULTS: A normal distribution of T2 was detected in resting muscles at 34 ± 3 ms (mean ± standard deviation [SD]) in 26 subjects using the Kolmogorov-Smirnov test, the Shapiro-Wilk test, and the Jarque-Bera test (P > 0.05). No gender differences were shown between the T2 or SD, and a similar result was obtained in 12 measurements on a single subject (P < 0.01). The T2r and SDr were used for reference values. The threshold ZT = 1.00 showed the highest sensitivity (0.86) even with 5% MVC, but it showed a lower specificity (0.85) than the other thresholds. ZT = 3.07 showed the highest specificity (1.0), but it showed a lower sensitivity (0.36) with the 5% MVC, compared with ZT = 2.56 (0.50). The receiver operating characteristics analysis also supported these results. CONCLUSION: We found that the T2 distribution in muscles was Gaussian, suggesting that a one-sample t-test can be applied, and that ZT = 2.56 could cover low-intensity exercise with high specificity and a low false-positive rate.

Roles of Keber's valve and foot chamber for foot manipulation in the mussel Nodularia douglasiae.

In order to analyse the roles of Keber's valve for foot manipulation in the mussel Nodularia douglasiae, the anatomy and haemolymph flow in the cardiovascular system were detected by magnetic resonance imaging. The superficial layer of the foot was covered by a dense muscle layer, which extended to the dorsal side and connected with the shell. This closed space, the foot chamber, had an inlet (anterior aorta) and an outlet (Keber's valve). At rest, in the beginning of the systolic phase, flows in the anterior aorta and the pedal artery increased, followed by the pedal and visceral sinuses. Then these flows ceased at the end of the systolic phase, followed by inflow to the ventricle in the diastolic phase; therefore, the compliance of the foot chamber is low enough to transfer pressure pulses to the visceral sinus. Extension of the foot started with relaxation of the foot muscle, so the compliance of the foot chamber increased. Then, Keber's valve closed so that the haemolymph filled the foot haemocoel. Retraction of the foot is initiated by the opening of Keber's valve. Judging from these results Keber's valve and the foot chamber are essential for circulation at rest, foot extension and retraction.This article has an associated First Person interview with the first author of the paper.

Accumulation and excretion of manganese ion in the kidney of Mytilus galloprovincialis.

T1-weighted magnetic resonance imaging (T1w-MRI) was employed to detect the accumulation of manganese ion (Mn2+) in urine in the kidney of the mussel Mytilus galloprovincialis, and the longitudinal relaxation rates (1/T1=R1) were measured. When the mussel was exposed to seawater containing 10 µmol l-1 Mn2+, the T1w-MRI intensity and R1 of the kidney, stomach and digestive glands were increased. Mn2+ might be taken into the hemolymph via the gastrointestinal tract, and then filtrated into the pericardium via the auricles. Although the image intensity in the pericardium was not affected by manganese, an image intensity enhancement was observed in the distal part of the renopericardial communication canals between the pericardium and the kidneys, indicating Mn2+ was concentrated in the excretion pathway. As the seawater Mn2+ concentration ([Mn2+]SW) was increased from 3 to 50 µmol l-1, R1 of the kidney (R1K) was elevated. When the mussels were immersed in 3-10 µmol l-1 [Mn2+]SW for 24 h, the Mn2+ concentration in the kidney ([Mn2+]K) showed a 15-fold increase compared with the ambient [Mn2+]SW In the range of [Mn2+]SW from 10 to 50 µmol l-1, R1K reached a plateau level that corresponded to 200 µmol l-1 [Mn2+]K As [Mn2+]K fell transiently, voluntary excretion of urine from the kidney was assumed. The decreases in intensity were not synchronized between the right and left kidneys, and the closure of the shells might not be essential for urinary excretion. The voluntary excretion suggested an additional explanation for the large range in metal concentratons in the kidneys of the mussel.

Repeated mild shaking of neonates induces transient cerebral microhemorrhages and anxiety-related behavior in adult rats.

Growing evidence suggests that neonatal cerebral microhemorrhages (MHs) are implicated in neuropsychiatric diseases in adults. Although animal studies have identified the progression of the underlying mechanisms of MHs, few studies have investigated the histopathology and behavioral outcomes. In this study, we created an experimental rat model of MHs using a new experimental device for repeated mild shaking brain injury (SBI) in the neonatal period and examined temporal changes in MHs using susceptibility weighted imaging (SWI) and iron histochemistry. SWI demonstrated transient MHs in the gray matter of the cerebral cortex and hippocampus in injured rats. Iron histochemical staining demonstrated leakage of iron and iron-positive cells surrounding MHs. This staining pattern lasted for a long time and continued after disappearance of hemorrhagic signals on SWI. These data suggested the presence of iron-associated gray matter injury after MHs. In the open field test, these injured rats showed anxiety-related behavior as adults. This model may be useful for exploring the underlying mechanisms of changes that occur after MHs and the behavioral outcomes of repeated mild SBI in early development.

Changes in T2-weighted MRI of supinator muscle, pronator teres muscle, and extensor indicis muscle with manual muscle testing.

[Purpose] In order to detect muscle activity with manual muscle testing, T2-weighted magnetic resonance (T2w-MR) images were detected by a 0.2 T compact MRI system. [Subjects and Methods] The subjects were 3 adult males. Transverse T2-weighted multi-slice spin-echo images of the left forearm were measured by a 39 ms echo-time with a 2,000 ms repetition time, a 9.5 mm slice thickness, 1 accumulation and a total image acquisition time of 4 min 16 s. First, T2w-MR images in the resting condition were measured. Then, manipulative isometric contraction exercise (5 sec duration) to the supinator muscle, the pronator teres muscle or the extensor indicis muscle was performed using Borg's rating of perceived exertion (RPE) scale of 15-17. The T2w-MR images were measured immediately after the exercise. [Results] T2w-MR image intensities increased significantly in the supinator muscle, the pronator teres muscle and the extensor indicis muscle after the exercise. However, the image intensities in the rest of the muscle did not change. [Conclusion] Using T2w-MR images, we could detect muscle activity in a deep muscle, the supinator muscle, and a small muscle, the extensor indicis muscle. These results also support the reliability of the manual muscle testing method.

A portable infrared photoplethysmograph: heartbeat of Mytilus galloprovincialis analyzed by MRI and application to Bathymodiolus septemdierum.

Infrared photoplethysmogram (IR-PPG) and magnetic resonance image (MRI) of the Mytilus galloprovincialis heart were obtained simultaneously. Heart rate was varied by changing temperature, aerial exposure and hypoxia. Higher heart rates (35-20 beat min-1) were usually observed at 20°C under the aerobic condition, and typical IR-PPG represented a single peak (peak v). The upward and downward slopes of the peak v corresponded to the filling and contracting of the ventricle, respectively. A double-peak IR-PPG was observed in a wide range of heart rates (5 to 35 beats min-1) under various conditions. The initial peak v corresponded to the filling of the ventricle, and the origin of the second peak (v') varied with the heart rate. A flat IR-PPG with a noise-level represented cardiac arrest. Although large movement of the shells and the foot caused slow waves or a baseline drift of the IR-PPG, the heart rate can be calculated from the v-v interval. Based on these results, we assembled a portable IR-PPG recording system, and measured the heartbeats of Bathymodiolus septemdierum (Mytilidae) for 24 h on a research vessel just after sampling from the deep sea, showing that IR-PPG is a noninvasive, economical, robust method that can be used in field experiments.

Involvement of myristoylated alanine-rich C kinase substrate phosphorylation and translocation in cholecystokinin-induced amylase release in rat pancreatic acini.

Cholecystokinin (CCK) is a gastrointestinal hormone that induces exocytotic amylase release in pancreatic acinar cells. The activation of protein kinase C (PKC) is involved in the CCK-induced pancreatic amylase release. Myristoylated alanine-rich C kinase substrate (MARCKS) is a ubiquitously expressed substrate of PKC. MARCKS has been implicated in membrane trafficking in several cell types. The phosphorylation of MARCKS by PKC results in the translocation of MARCKS from the membrane to the cytosol. Here, we studied the involvement of MARCKS in the CCK-induced amylase release in rat pancreatic acini. Employing Western blotting, we detected MARCKS protein in the rat pancreatic acini. CCK induced MARCKS phosphorylation. A PKC-δ inhibitor, rottlerin, inhibited the CCK-induced MARCKS phosphorylation and amylase release. In the translocation assay, we also observed CCK-induced PKC-δ activation. An immunohistochemistry study showed that CCK induced MARCKS translocation from the membrane to the cytosol. When acini were lysed by a detergent, Triton X-100, CCK partially induced displacement of the MARCKS from the GM1a-rich detergent-resistant membrane fractions (DRMs) in which Syntaxin2 is distributed. A MARCKS-related peptide inhibited the CCK-induced amylase release. These findings suggest that MARCKS phosphorylation by PKC-δ and then MARCKS translocation from the GM1a-rich DRMs to the cytosol are involved in the CCK-induced amylase release in pancreatic acinar cells.

Changes to Muscle T2after Single-finger Exercise Measured with 0.2T MR Imaging.

We constructed an arm holder for muscle exercise from a forearm-shaped plastic shell and magnetic resonance (MR) imaging position markers and determined the echo time (39 ms) for T2-weighted spin-echo MR imaging from T2values of the exercised (50 ms) and resting (32 ms) muscle at 0.2 tesla. The smallest detectable muscle was the extensor digiti minimi muscle (cross-sectional area 25 mm²). This combination could be useful to monitor finger exercise in patients undergoing physical therapy.

Structure and Size-selective Permeability of the Synovial Membrane of the Temporomandibular Joint of the Mouse Measured by MR Imaging at 7T.

PURPOSE: We analyzed the anatomical structure of the temporomandibular joint (TMJ) and molecular weight dependency of synovial membrane permeability in mice using 7-tesla magnetic resonance (MR) imaging. METHODS: We obtained 3-dimensional (3D) T1-weighted gradient echo (3D-T1W) and 3D T2-weighted rapid acquisition with relaxation enhancement (3D-T2W RARE) MR images of the TMJ of male C57BL6 mice with voxel resolution of 65 µm. Two-dimensional (2D) T1w images were measured every 45 s before and after bolus intravenous (IV) injection of contrast reagents: gadolinium diethylenetriamine pentaacetic acid (Gd-DTPA; 0.5 kDa); oligomer-based contrast agent (CH3-DTPA-Gd; 2.1 kDa); gadolinium-labeled polylysine (Gd-polylysine; 10 kDa); and gadolinium-labeled albumin (Gd-albumin; 74 kDa). RESULTS: T1W images depicted the temporal bone and mandibular condyle as regions with lower signal intensity and the disc as a region of intermediate intensity. In the Gd-DTPA-enhanced T1W and T2W images, the articular disc could be identified as a region with lower signal intensity than that of the upper and lower joint cavities. After IV injection of Gd-DTPA or CH3-DTPA-Gd, the signal intensity of the joint cavities increased within 10 min, but this increase was not shown with Gd-polylysine and Gd-albumin. CONCLUSION: The structural findings obtained by MR imaging agreed with those obtained by hematoxylin-eosin staining under light microscopy. Contrast-enhanced MR imaging suggested that smaller (<2.1 kDa) but not larger (>10 kDa) molecules can permeate the synovial membrane. Our results suggest the utility of MR imaging for analyzing the structure of the TMJ as well as permeability of the synovial membrane.