Distribution of the axillary nerve to the subacromial bursa and the area around the long head of the biceps tendon.

PURPOSE: Patients with a shoulder disorder often complain of pain on the anterior or lateral aspect of the shoulder. Such pain has been thought to originate from the suprascapular nerve. However, taking into consideration the distinctive course of the axillary nerve, the axillary nerve is likely to supply branches to the structure around the shoulder joint. This study was conducted to clarify the division, course, and distribution of the branches which originate from the axillary nerve and innervate structures around the shoulder joint. METHODS: The division, course, and distribution of the branches which originate from the axillary nerve and innervate structures around the shoulder joint were examined macroscopically by dissecting 20 shoulders of 10 adult Japanese cadavers. RESULTS: The thin branches from the anterior branch of the axillary nerve were distributed to the subacromial bursa and the area around the long head of the biceps tendon. The branches from the main trunk of the axillary nerve or the branch to the teres minor muscle were distributed to the infero-posterior part of the shoulder joint. CONCLUSION: The pain on the anterior or lateral aspect of the shoulder, which has been thought to originate from the suprascapular nerve, might be related to the thin branches which originate from the axillary nerve and innervate the subacromial bursa and the area around the long head of the biceps tendon. CLINICAL RELEVANCE: These results would be useful to consider the cause of the shoulder pain or to prevent the residual pain after the biceps tenodesis.

An artery accompanying the sciatic nerve (arteria comitans nervi ischiadici) and the position of the hip joint: a comparative histological study using chick, mouse, and human foetal specimens.

Birds and reptiles always carry a long and thick artery accompanying the sciatic nerve (i.e., the sciatic artery), whereas mammals do not. We attempted to demonstrate a difference in courses of the nerve and artery in fetuses in relation with the hip joint posture. Eight mid-term human fetuses (15-18 weeks), five mouse fetuses (E18) and five chick embryos (11 days after incubation) were examined histologically. Thin feeding arteries in the sciatic nerve were consistently observed in human fetuses in spite of the long, inferiorly curved course of the nerve around the ischium. The tissue around the human sciatic nerve was not so tight because of the medial and inferior shift of the nerve away from the hip joint. The fetal hip joint position differed among the species, being highly flexed in humans and almost at right angle flexion in mice and chicks. Because of deep adduction of the hip joint in the mouse, the knee was located near the midline of the body. The mouse sciatic nerve ran through the tight tissue along the head of the femur, whereas the chick nerve ran through the loose space even in the gluteal region. In birds, evolution of the pelvis including the hip joint without adduction seemed to make the arterial development possible. In mammals, highly flexed or adducted hip joint seemed to be one of the disturbing factors against development of the long and thick artery. A slight change in posture may cause significant arterial variation.

An anatomic study of structure and innervation of the serratus anterior muscle.

BACKGROUND: The structure and function of the serratus anterior muscle are partitioned into three parts. If the morphological characteristics in each part can be demonstrated in more detail, the cause of dysfunction will probably be identifiable more accurately. The purpose of this study was to demonstrate the details of the structure and innervation in each part of the serratus anterior muscle. MATERIALS AND METHODS: This macroscopic anatomic study was conducted using ten sides from five cadavers. The structure and innervation in each part of this muscle were examined. RESULTS: In the superior part, the independent branch was divided from a branch innervating the levator scapulae muscle. In the middle part, the long thoracic nerve descended on one-third of the anterior region between the origin and insertion. In the inferior part, the long thoracic nerve which ramified into many branches and branches from the intercostal nerves were distributed on all sides. CONCLUSION: This study demonstrated that the innervation of the serratus anterior muscle was different in each part. The difference indicates that the superior part has an intimate relation with the levator scapulae muscle while the middle and inferior parts could be the actual serratus anterior muscle. Moreover, the distribution of branches from the intercostal nerves shows that the inferior part has a connection with some trunk elements. Understanding these characteristics of innervation is useful to identify the cause of dysfunction. In addition, we assert that the constant distribution of branches from the intercostal nerves is significant for the morphology.

Time-resolved optical mammography and its preliminary clinical results.

We have been developing an optical mammography prototype consisting of a multi-channel time-resolved spectroscopy system for breast cancer screening. The system utilizes the time-correlated single photon counting method, and the detector modules and the signal processing circuits were custom-made to obtain a high signal to noise ratio and high temperature stability with a high temporal resolution. Pulsed light generated by a Ti: Sapphire laser was irradiated to the breast, and the transmitted light was collected by optical fibers placed on the surface of a hemispherical gantry filled with an optical matching fluid. To reconstruct a 3D image of the breast, we employed a method using a time-resolved photon path distribution based on the assumption that scattering and absorption are independent of each other. We verified the possibility of human breast imaging by using a three-dimensional phantom model, which provides a simulation of human breast cancer, in the gantry. The clinical study was also started in January 2007. In a comparative study with conventional modalities, the breast cancers were detected as regions of optically higher absorption. Moreover, the results suggest that optical mammography is useful in monitoring the effects of chemotherapy.

Temperature effects on the development of Enteromyxum spp. (Myxozoa) in experimentally infected tiger puffer, Takifugu rubripes (Temminck & Schlegel).

The effects of water temperature on the development of the enteric myxosporeans, Enteromyxum fugu and Enteromyxum leei, were investigated in experimentally infected tiger puffer, Takifugu rubripes. After naïve tiger puffer were fed gut tissue infected with both E. fugu and E. leei, they were divided into separate tanks and kept at different constant temperature regimes between 10 and 25 degrees C. Regardless of the water temperature tested, E. fugu was consistently detected with a high prevalence of infection (60-100%), although no sporulation occurred at 10 and 15 degrees C. Development of E. leei and the onset of disease were suppressed by low water temperatures (<15 degrees C). However, a temperature increase to 20 degrees C promoted the development of E. leei, followed by an increase of disease rate in the fish. The present study demonstrates that water temperatures below 15 degrees C have an inhibitory effect on the development of E. fugu and E. leei, resulting in suppression of enteromyxosis at low temperatures.

Pheochromocytoma of the urinary bladder revealed with cerebral hemorrhage.

The case was a 51-year-old man, who has been undergoing treatment with oral medication for hypertension for three years. The patient was admitted to the author's clinic for hemorrhage in the left putamen. He was diagnosed as having primary pheochromocytoma of the bladder from such symptoms as paroxysmal blood pressure elevation after urination, mild increase in catecholamine levels before and after urination, and from the results of 131I-MIBG scintigraphy, and cystoscopy, and underwent excision of the bladder tumor. Upon endocrinological examination, only mild increases in catecholamine levels were found. Therefore, constant monitoring of blood pressure and 131I-MIBG scintigraphy were useful for a definitive diagnose.

Filamentous phage associated with recent pandemic strains of Vibrio parahaemolyticus.

A group of pandemic strains of Vibrio parahaemolyticus has recently appeared in Asia and North America. We demonstrate that a filamentous phage is specifically associated with the pandemic V. parahaemolyticus strains. An open reading frame unique to the phage is a useful genetic marker to identify these strains.

Elevated serum soluble CD44 level in sarcoidosis.

Sarcoidosis is a chronic multi-organ granulomatous disease of unknown etiology. Several studies have suggested an involvement of immunologic background in sarcoidosis. The lymphocyte surface marker CD44 is a multifunctional molecule which mediates the adhesion of lymphocytes to the extracellular matrix. Recently, we developed a system to quantitate soluble CD44 (sCD44) which we employed to determine serum and bronchoalveolar lavage fluid (BALF) levels of sCD44 to obtain further insights into immunologic aspects of sarcoidosis. Serum sCD44 levels were measured in 13 consecutive patients with sarcoidosis and 56 normal healthy controls using enzyme-linked immunoabsorbent assay. BALF sCD44 levels were also measured in 11 patients with sarcoidosis and 10 normal healthy controls. In patients with sarcoidosis, the serum sCD44 level was significantly higher than that of normal controls (348.5+/-164.2 ng/ml vs 145.4+/-22.9 ng/ml; p<0.001). Also BALF sCD44 levels tended to be higher in sarcoidosis than in normal controls (23.7+/-13.4 ng/ml vs 18.1+/-8.4 ng/ml), but no statistically significant difference was recognized. We also found that there was a positive correlation between the serum sCD44 and angiotensin converting enzyme (r=0.78). Our data indicate that sCD44 may be related to immunologic background and may be a useful new marker of sarcoidosis.

Orientations and locations of local anesthetics benzocaine and butamben in phospholipid membranes as studied by 2H NMR spectroscopy.

This paper presents experimental evidence that an aromatic compound that has a quadrupole moment locates in a polar headgroup region in the lipid membranes, but not in a membrane interior hydrophobic region, and discusses correlation to the site of action of benzocaine and butamben on sodium channels. The 2H NMR spectra of benzocaine-d4, benzocaine-d5, butamben-d4, and butamben-d9 in the model membranes were observed. The 2H NMR spectra of perdeuterated palmitic acid and selectively deuterated palmitic acids at C2, C3, C5, C6, C9, or C10, which were inserted into the lipid membranes, were also observed. The phosphatidylserine (PS), phosphatidylcholine (PC), and liquid mixtures composed of PS, PC, and phosphatidylethanolamine (PE), which contain or do not contain cholesterol, were employed. A moment analysis was applied to the 2H NMR spectra of palmitic-d31 acid. An order parameter, S(CD), for each carbon segment was calculated from the observed quadruple splitting. We concluded that in the lipid mixture containing cholesterol, the aromatic rings of benzocaine and butamben locate around the glycerol moiety of the lipids and that when there exists no cholesterol, they locate a little more inside from the headgroup region, directing, in both cases, their amino groups upward (polar region) and the ethyl and butyl groups downward (hydrophobic region). These data cast a question on the site of action of the neutral local anesthetics in the sodium channels.

A filamentous phage associated with recent pandemic Vibrio parahaemolyticus O3:K6 strains.

A specific serotype, O3:K6, of Vibrio parahaemolyticus has recently been causing epidemics of gastroenteritis in Southeast Asia, Japan, and North America. To examine whether the new O3:K6 strains possess characteristics that may exacerbate outbreaks, we compared V. parahaemolyticus O3:K6 strains with non-O3:K6 strains using strains isolated from individuals with traveler's diarrhea at Kansai Airport Quarantine Station, Osaka, Japan. All 24 O3:K6 strains possessed a common plasmid, pO3K6 (DNA size, 8,782 bp, with 10 open reading frames [ORFs]). The gene organization of pO3K6 was similar to that of Vf33, a filamentous phage previously described in V. parahaemolyticus. We isolated a phage (phage f237) from the culture supernatant of V. parahaemolyticus O3:K6 strain KXV237, which formed a turbid plaque on an indicator strain. The genome of f237 was single-stranded DNA, and the double-stranded DNA obtained by treatment of the genome with DNA polymerase was identical to that of pO3K6 when analyzed by agarose gel electrophoresis after HindIII digestion. Furthermore, the N-terminal amino acid sequence of the f237 major coat protein was found in ORF4 of pO3K6. Our results showed that pO3K6 is a replicative form of f237. Among the ORFs found in the f237 genome, the sequence of ORF8 had no significant homology to those of any proteins in databases. ORF8 was located on a region corresponding to the distinctive region of Vf33, and its G+C content was apparently lower than that of the remaining DNA sequence of f237. By colony hybridization, ORF8 was detected only in O3:K6 strains isolated since 1996 and was not found in O3:K6 strains isolated before 1996 and clinical V. parahaemolyticus strains other than those of serotype O3:K6. Thus, this study shows that f237 is exclusively associated with recent V. parahaemolyticus O3:K6 strains. The ORF8 gene can be a useful genetic marker for the identification of the recently widespread O3:K6 strains of V. parahaemolyticus.

mkp-1 encoding mitogen-activated protein kinase phosphatase 1, a verotoxin 1 responsive gene, detected by differential display reverse transcription-PCR in Caco-2 cells.

The major cytotoxic effect of the verotoxins (VTs) produced by strains of VT-producing Escherichia coli is the inhibition of host-cell protein synthesis, but VTs are also suspected to play a role in apoptotic cell signaling and cytokine release. Four differentially expressed genes, including mkp-1 (encoding mitogen-activated protein kinase phospatase 1), were detected by differential display reverse transcription-PCR (DD RT-PCR) stimulated by VT1 in Caco-2 cells. Northern blot analysis showed the induction of mkp-1 mRNA 6 h after VT1 stimulation. Neither mutant VT1 (mutVT1), harboring two mutations in the A subunit (E167Q-R170L), nor cycloheximide induced mkp-1 mRNA, but mkp-1 mRNA was detected with both wild-type VT1 (wtVT1) and anisomycin, a 28S rRNA inhibitor. Therefore, we concluded that the A subunit of VT1 was essential for mkp-1 induction. Increased amounts of phosphorylated c-Jun protein were also found with wtVT1 and anisomycin. Although the precise mechanism of induction of MKP-1 is unknown, we hypothesized that 28S rRNA not only was a sensor for ribotoxic stress, but also was involved in the signal cascade of MKP-1. This is the first report of detection by DD RT-PCR of cellular genes induced by bacterial toxins.

[Marked airway constriction due to relapsing polychondritis].

A 40-year-old woman was admitted to our hospital presenting with dry cough, sore throat, and fever. Her right auricle was markedly swollen and her ocular conjunctiva were reddened, suggesting scleritis. A chest computed tomogram showed narrowing of the trachea due to enlarged mediastinal soft tissues. These clinical findings suggested the patient had relapsing polychondritis. A cartilage biopsy from her right auricle was taken to confirm the diagnosis. Pathological findings revealed loosening and dissolution of cartilage and infiltration of lymphocytes, which were consistent with relapsing polychondritis. Although prednisolone was given to the patient, her symptoms were not alleviated. Methylprednisolone pulse therapy and oral diaminodiphenylsulfone were added to the patient's treatment regimen. With this combination, her symptoms gradually subsided. Constriction of the airway has been cited as one of the prognostic factors in relapsing polychondritis. In view of the danger of sudden death caused by airway obstruction, close observation of the patient in this case was considered necessary.

Serum soluble CD44 levels for monitoring disease states in acute leukemia and myelodysplastic syndromes.

To determine the clinical implications of soluble CD44 (sCD44) levels in hematologic neoplasias, we developed an enzyme-linked immunosorbent assay for sCD44 using two monoclonal antibodies to the standard 90 kDa form, and assessed the serum concentration of sCD44 in normal healthy volunteers, patients with acute leukemia, myelodysplastic syndromes (MDS), and those with chronic myeloid leukemia (CML). Compared to that in normal individuals (n=51; 145. 1 24.6 ng/ml), the serum sCD44 level was significantly elevated in patients with acute myeloid leukemia (AML; n=18; 331.9 99.0 ng/ml, P=0.0001), acute lymphoid leukemia (ALL; n=16; 551.3 427.8 ng/ml, P=0.0001) and CML (n=18; 262.0 97.5 ng/ml, P=0.0001). The sCD44 level was slightly elevated in patients with MDS (n=43; 173.8 54.9 ng/ml, P=0.0071). In patients with acute leukemia, serum sCD44 concentrations decreased significantly in response to treatment and reached nearly normal levels after complete remission (P=0.0005 in AML and P=0.0032 in ALL). The sCD44 levels in patients with MDS increased after they developed acute leukemia, whereas no significant difference in sCD44 levels was observed between the chronic and the blastic phases in patients with CML. Our results indicate that serum sCD44 levels may be a useful marker for monitoring response to treatment and disease progression, especially in acute leukemia.

Serum concentrations of free ubiquitin and multiubiquitin chains.

Ubiquitin, which can conjugate with cellular proteins, is classified into two forms: free ubiquitin and multiubiquitin chains. The latter is active as a signal for degradation of the targeted proteins. We found both forms in human serum and, using two immunoassays, quantitated them in sera from healthy subjects and patients with some diseases. Because of putative leakage of erythrocyte ubiquitin, hemolytic serum and serum obtained after long incubation (> 1-2 h) of blood at room temperature were excluded. Serum concentrations of multiubiquitin chains and free ubiquitin were substantially higher in rheumatoid arthritis and hemodialysis patients, respectively, than healthy subjects. Additionally, in acute viral hepatitis, serum multiubiquitin chain concentrations were increased in the acute phase, decreased in the recovery phase, and correlated with alanine and aspartate aminotransferase activities (r = 0.676 and 0.610, P < 0.0001 and < 0.001, respectively). Therefore, serum ubiquitin may have prognostic value.

Malignant aortic body tumor in a Holstein cow.

A malignant aortic body tumor was observed in a 5-year-old female Holstein cow. The neoplastic mass, of 22 x 17 x 15 cm in size, was located at the base of the left atrium, having irregular lobular structures. The tumor cells had slightly eosinophilic cytoplasm, and a round or oval nucleus. Metastasis was only present in the premediastinal lymph node. The tumor cells exhibited intense immunoreactivity for neuron-specific enolase (NSE) and synaptophysin, and were moderately positive for chromogranin A. Electronmicroscopy revealed membrane-limited granules in the cytoplasm. The cultured cells were spindle in shape, and having projectional cytoplasm. They were intensely positive for NSE, synaptophysin, chromogranin A, and neurofilament (200 kD). Consequently, this case was diagnosed as a malignant aortic body tumor from the neuroecrodermal origin.

Locations of local anesthetic dibucaine in model membranes and the interaction between dibucaine and a Na+ channel inactivation gate peptide as studied by 2H- and 1H-NMR spectroscopies.

To study the molecular mechanisms of local anesthesia, locations of local anesthetic dibucaine in model membranes and the interactions of dibucaine with a Na+ channel inactivation gate peptide have been studied by 2H- and 1H-NMR spectroscopies. The 2H-NMR spectra of dibucaine-d9 and dibucaine-d1, which are deuterated at the butoxy group and at the 3 position in its quinoline ring, respectively, have been observed in multilamellar dispersions of the lipid mixture composed of phosphatidylcholine, phosphatidylserine, and phosphatidylethanolamine. 2H-NMR spectra of deuterated palmitic acids incorporated, as a probe, into the lipid mixture containing cholesterol have also been observed. An order parameter, SCD, for each carbon segment was calculated from the observed quadrupole splittings. Combining these results, we concluded that first, the butoxy group of dibucaine is penetrating between the acyl chains of lipids in the model membranes, and second, the quinoline ring of dibucaine is located at the polar region of lipids but not at the hydrophobic acyl chain moiety. These results mean that dibucaine is situated in a favorable position that permits it to interact with a cluster of hydrophobic amino acids (Ile-Phe-Met) within the intracellular linker between domains III and IV of Na+ channel protein, which functions as an inactivation gate. To confirm whether the dibucaine molecule at the surface region of lipids can really interact with the hydrophobic amino acids, we synthesized a model peptide that includes the hydrophobic amino acids (Ac-GGQDIFMTEEQK-OH, MP-1), the amino acid sequence of which corresponds to the linker part of rat brain type IIA Na+ channel, and the one in which Phe has been substituted by Gln (MP-2), and measured 1H-NMR spectra in both phosphate buffer and phosphatidylserine liposomes. It was found that the quinoline ring of dibucaine can interact with the aromatic ring of Phe by stacking of the rings; moreover, the interaction can be reinforced by the presence of lipids. In conclusion, we wish to propose that local anesthesia originates from the pi-stacking interaction between aromatic rings of an anesthetic molecule located at the polar headgroup region of the so-called boundary lipids and of the Phe in the intracellular linker between domains III and IV of the Na+ channel protein, prolonging the inactivated state and consequently making it impossible to proceed to the resting state.

Rapid analysis of four bilirubins in domestic animal sera using high-performance liquid chromatography.

A rapid method was developed to analyze delta-bilirubin (B delta), diconjugated bilirubin (DCB), monoconjugated bilirubin (MCB), and unconjugated bilirubin (Bu) by direct injection of sera using high-performance liquid chromatography (HPLC) with an internal-surface reversed-phase silica support (ISRP) column. Sharp bilirubin peaks were obtained using a simple mobile phase of acetonitrile: 0.5 M Tris-HCl buffer (20:80, v/v, pH 7.2). A variable-wavelength detector set at 450 nm, 0.01 absorbance unit full scale (AUFS), and a recorder set at 4 mm/min were used for detection. Peaks for B delta, DCB, MCB and Bu appeared at 4.4, 6.4, 9.2 and 14.5 min, respectively, in human serum from subject with obstructive jaundice which was used as a bilirubin standard throughout this experiment. The mean recovery rate after direct addition of Bu in swine serum was 91.9% and that of DCB was 95.9%. When sera from icteric cattle, pigs and horses were analyzed using the direct injection technique, four bilirubin peaks were obtained and there was reliable correlation between the sum of the bilirubin peak heights observed on HPLC and the total bilirubin value measured by a standard reference procedure.

Rapid and quantitative analysis of bilirubin in equines by high-performance liquid chromatography.

Rapid and quantitative analytical methods for bilirubin using high-performance liquid chromatography (HPLC) with UV detection were developed for samples from equines at a meat inspection site. Sharp HPLC peaks for bilirubins, unconjugated bilirubin (UCBL) and conjugated bilirubin (CBL), were obtained using a simple mobile phase of methanol:0.5 M Tris-HCl buffer (65:35, v/v, pH 7.4). A variable wavelength detector set at 450 nm, 0.01 AUFS and a recorder set at 4 cm/min were used for detection. Peaks for UCBL and CBL occurred at 7.1 min and 4.9 min, the lower limits of detection ranged between 0.16 and 0.78 microgram/ml, respectively. Orange II, for which the retention time was 3.6 min, was selected as an internal standard. When the samples were analysed from healthy equines and those suspected of being jaundiced due to a yellow colour change in the carcasses, only UCBL peaks were recognized, and CBL peaks were never obtained. Aqueous humour is a very common sample at meat inspections but UCBL or CBL peaks were all absent in both healthy and clinical samples from equines. There were reliable correlations between UCBL peaks (microV) using HPLC and total bilirubin value (mg/dl) measured by the total bilirubin assay kit in serum. A correlation between peak height (microV) on HPLC from serum samples and that of pericardial fluid samples was obtained.