Direct comparison of anti-inflammatory effects of 14-, 15-, and 16-membered macrolide antibiotics in experimental inflammation model induced by carrageenan in rats.

Some macrolide antibiotics, which share a basic lactone ring structure, also exhibit anti-inflammatory actions in addition to their antibacterial activities. However, no study has directly compared anti-inflammatory effects on acute inflammation among macrolide antibiotics with the distinct size of the lactone ring. In this study, we evaluated and compared the anti-inflammatory activities of four 14-membered macrolides (erythromycin, clarithromycin, roxithromycin, oleandomycin), one 15-membered macrolide (azithromycin), and three 16-membered macrolides (midecamycin, josamycin, leucomycin) using a rat carrageenan-induced footpad edema model. All macrolide antibiotics were intraperitoneally administered to rats one hour before the induction of inflammatory edema with 1% λ -carrageenan. The anti-inflammatory effects on acute inflammation were evaluated by changing the edema volume. All 14-membered and 15-membered macrolide antibiotics significantly suppressed the development of edema. Conversely, none of the 16-membered macrolide antibiotics inhibited the growth of edema. In conclusion, compared to 16-membered macrolide antibiotics, 14-membered and 15-membered macrolide antibiotics have stronger anti-inflammatory effects. Further research should be done to determine why different lactone ring sizes should have distinct anti-inflammatory effects.

Anti-edematous effects of epinastine, cetirizine and its enantiomers in λ-carrageenan-induced edema in rat hind paw.

Urticaria is induced by the histamine released from mast cells which develops wheals (edema) as a visual feature. In clinical practice, second-generation histamine H1 -receptor blockers are routinely used as the first-line symptomatic treatment for urticaria. Nevertheless, not much research has directly examined the second-generation histamine H1-receptor blockers' ability to reduce edema. In this study, we directly evaluated the anti-edematous activities of three second-generation histamine H1-receptor blockers available in the market (epinastine hydrochloride, cetirizine hydrochloride, and levocetirizine hydrochloride) using a λ-carrageenan-induced footpad edema model. One hour before the induction of edema with 1% λ -carrageenan injection, all second-generation histamine H1 -receptor blockers (5, 10, 50 and 100 mg/kg) were subcutaneously administered to rats. At 0.5 and 3 hours after λ -carrageenan administration, the edema volume was evaluated using a Plethysmometer. Epinastine hydrochloride significantly suppressed the edema growth in a dose-dependent manner. Cetirizine hydrochloride showed a slight anti-edematous effect, while levocetirizine significantly inhibited the development of edema in a dose-dependent manner. On the other hand, dextrocetirizine did not prevent edema from growing. In summary, second-generation histamine H1 -receptor blockers, at least those examined in this study, may be able to reduce the clinical symptoms of urticaria associated with edema. Levocetirizine hydrochloride is also anticipated to have stronger anti-edematous effects than cetirizine hydrochloride because levocetirizine is responsible for cetirizine's anti-edematous activity.

Anti-Inflammatory Actions of Expectorants in a Rat Carrageenan-Induced Footpad Edema Model.

S-Carboxymethyl-L-cysteine (SCMS) exhibits sputum-regulating and anti-inflammatory actions. Previous studies reported the anti-inflammatory effects of SCMS on chronic inflammatory diseases, but no study has examined these effects on acute inflammatory diseases. In this study, we investigated the anti-inflammatory effects of SCMS in a rat carrageenan-induced footpad edema model, which is routinely used as an acute inflammation model. Expectorants were administered to rats with footpad edema induced by subcutaneously administering 1%λ-carrageenan to the footpad of the left posterior limb, and the dose dependency of the anti-inflammatory effects was evaluated. As a result, even when the dose of SCMS was increased to 400 mg/kg, there were no inhibitory effects on edema. Furthermore, we examined the inhibitory effects of other expectorants (ambroxol hydrochloride, N-acetyl-L-cysteine, L-cysteine ethylester hydrochloride, and L-cysteine methylester hydrochloride), which were reported to exhibit anti-inflammatory effects on chronic inflammation, on edema. However, none of these expectorants inhibited edema.

Proteomics-based identification of novel proteins in temporal tendons of patients with masticatory muscle tendon--aponeurosis hyperplasia.

Masticatory muscle tendon-aponeurosis hyperplasia (MMTAH) is a new disease associated with limited mouth opening that is often misdiagnosed as a temporomandibular disorder; subsequently, patients are mistakenly treated with irreversible operations. Due to the poor presentation and characterization of symptoms, the underlying pathological conditions remain unclear. We have previously conducted a proteomic analysis of tendons derived from one MMTAH subject and one facial deformity subject using two-dimensional fluorescence difference gel electrophoresis and liquid chromatography coupled with tandem mass spectrometry. However, the results were obtained for only one subject. The aim of the present study was to confirm the expression of specific molecules in tendon tissues from multiple subjects with MMTAH by applying two-dimensional polyacrylamide gel electrophoresis with matrix-assisted laser desorption ionization time-of-flight mass spectrometry. Of the 19 proteins identified in tendons from both MMTAH and facial deformity patients, fibrinogen fragment D and beta-crystallin A4 were up-regulated, whereas myosin light chain 4 was down-regulated in MMTAH. We also found fibrinogen to be expressed robustly in tendon tissues of MMTAH patients. Our data provide the possibility that the distinctive expression of these novel proteins is associated with the pathology of MMTAH.

[Complete resection for giant thymic carcinoma after simultaneous combination chemotherapy].

A 19-year-old man visited our hospital complaining of dyspnea. Chest X-ray and computed tomography (CT) showed a huge mass in the right anterior mediastinum. We diagnosed this as invasive thymoma by microscopic examination of specimens obtained by echo-guided needle biopsy. The patient underwent 6 courses chemotherapy [1st course : carboplatin (CBDCA) + doxorubicin hydrochloride (DXR) + vincristine sulfate (VCR) + cyclophosphamide (CPA), 2nd, 3rd-6th course : cisplatin (CDDP) + ADM + VCR + CPA]. At achievement of partial response (the reduction rate of the tumor size : 91.4%), the tumor was completely resected. The pathological examination of the resected specimens yielded a diagnosis of large cell carcinoma. Preoperative chemotherapy with ADOC regimen may be effective in advanced thymic carcinoma.

Radial magnetic resonance imaging and pathological findings of acetabular labrum in dysplastic hips.

Radial MRI findings and pathological changes were comparatively examined in the acetabular labrum of 11 hips of 11 patients, who underwent total hip arthroplasty for osteoarthritis due to acetabular dysplasia. Diffuse high signal pattern on the radial MR images corresponded to histological degeneration of the labrum. High signal pattern which was equivalent to the synovial fluid, showed an intralabral tear. In the obscure areas of MR images, severe impairment of the labrum such as rupture, detachment, and displacement were found.

Gizzard myoglobin contents and feeding habits in avian species.

In an attempt to consider physiological function of myoglobin (Mb), we determined Mb contents of gizzard smooth muscles with special reference to feeding habits in 85 avian species of 19 orders. The Mb content in 44 species of herbivorous birds was 7.52+/-3.81 mg/g wet muscle, which was significantly higher than the value of 2.34+/-1.74 mg/g in 41 species of carnivorous ones (P<0.001). Buffering capacity, as determined by in vitro titration method, was 37.3+/-5.5 slykes/g in gizzard smooth muscles of 75 species and 60.7+/-10.5 slykes in breast skeletal muscles of 77 species (P<0.001), which suggests a significantly higher dependence, almost comparable to cardiac muscles, of the gizzard muscular function on aerobic metabolism. Together with the fact that blood circulation in the gizzard is very low at resting, and might be further limited during activity, we conclude that the higher Mb content in gizzards of herbivorous birds is an adaptation, to allow storage and/or facilitated diffusion of oxygen, during process of high mechanical work required to grind down hard and fibrous vegetable food under the conditions of limited circulatory supply.

[A case of atypical polypoid adenomyoma with intramural myoma-like growth].

We encountered a case of a large atypical polypoid adenomyoma (APA) which showed characteristic intramural appearance. As many cases of APA of the uterus have been reported in recent years, macro- and microscopic findings of the lesion and its clinical characteristics have been investigated. However the pathogenesis and the natural history of the lesion remained to be solved owing to still limited number of the cases. We report this case to discuss the macroscopic findings and clinicopathologic characteristics of the lesion.

Isolation and properties of myoglobins from rat (Rattus norvegicus) skeletal muscles.

One major (Mb I) and two minor (Mbs II and III) myoglobins, were isolated by a heat-denaturation-gel-filtration-chromato-focusing procedure from aqueous extract of rat skeletal muscles. The primary structure of the major component, as determined by an automatic Edman degradation method, revealed amino acid replacements at nine positions compared with murine myoglobin. As in murine myoglobin, a cysteine residue, highly reactive to p-chloromercuribenzoate (PMB), was present at position 66. The two minor components, with higher anodic mobilities and with SH group unreactive to PMB, were found to be derived from Mb I through mixed disulfide formation with cysteine (Mb II) or glutathione (Mb III) at the Cys66 residue. In view of the hydropathy profile, secondary structures of rat and mouse myoglobins showed few differences from each other. The three rat myoglobins displayed essentially identical oxygen equilibrium properties with neither homotropic nor heterotropic allosteric interactions, which agreed very well with those computed from the kinetic measurements.

A novel method for determination of alpha1,6fucosyltransferase activity using a reducing oligosaccharide from egg yolk as a specific acceptor.

A new method for determination of alpha1,6fucosyltransferase activity has been described. Recently, the disialyl-biantennary undecasaccharide was prepared in high yield from egg yolk [(1996), Carbohydr Lett 2: 137-42]. By treatment of this oligosaccharide with neuraminidase and beta-galactosidase, we readily obtained an asialo-agalacto-biantennary heptasaccharide (GlcNAcbeta 1,2Manalpha1,6[GlcNAcbeta1,2Manalpha1,3]Manbeta1 ,4GlcNAcbeta1,4GlcNAc). Using this asialo-agalacto-oligosaccharide as an acceptor, fucosyltransferases from human plasma and extracts of various human hepatoma cell lines were assayed in the presence of GDP-[3H]fucose. The reaction mixture was applied to a column of GlcNAc-binding, Psathyrella velutina lectin coupled gel. All the fucosylated acceptor were bound to the column which was eluted with 50 mM GlcNAc. Structural analyses revealed that only the innermost GlcNAc residue of the acceptor was fucosylated through an alpha1,6-linkage, and the oligosaccharide prepared could be used as a specific acceptor for alpha1,6fucosyltransferase. The present method was used to screen plasma alpha1,6fucosyltransferase in several patient groups, and significantly elevated activities were found in samples from patients with liver diseases, including chronic hepatitis, liver cirrhosis, and hepatocellular carcinoma.

Circulatory arrest with profound hypothermia during the surgical treatment of large internal carotid artery aneurysm--case report.

A 43-year-old male presented with a cerebral aneurysm manifesting as right facial paresthesia, without neurological deficit. Angiography revealed a large aneurysm (22 mm) of the left internal carotid artery. Intravascular treatment using placement of a detachable coil was attempted, but the coil did not stay in the aneurysmal cavity and the procedure was abandoned. The patient did not tolerate the transient balloon occlusion test of the left internal carotid artery. Therefore, the aneurysm was clipped through an open craniotomy with profound hypothermia (20 degrees C) with cardiac arrest (24 minutes). The aneurysmal dome was collapsed, allowing easy dissection of the posterior communicating artery. The closed chest method was used during the extracorporeal cardiopulmonary bypass. Postoperative angiography revealed complete neck clipping with preservation of carotid blood flow. The patient recovered well and resumed his employment. Circulatory arrest with hypothermia provides several benefits for the surgical treatment of large and giant aneurysms.

Oxygen uptake in maximally contracting muscle perfused at low flow with high O2 affinity erythrocytes.

To analyze the roles of blood flow and blood-tissue diffusion in determining oxygen (O2) availability to maximally exercising skeletal muscle, we investigated the influence of low flow on the O2 uptake (VO2)-venous PO2 (PVO2) relation with normal and high O2 affinity erythrocytes. We prepared normal and high O2 affinity erythrocytes by incubating human citrate-phosphate-dextrose-stored erythrocytes with and without cyanate. We perfused maximally contracting gastrocnemius muscle at 4-Hz isometric twitch with solutions containing normal and high O2 affinity erythrocytes for 3 min in anesthetized dogs. During stimulation, perfusion flow was maintained low at 50 ml.min-1.100 g-1 with a perfusion pressure of 50 mmHg. PVO2 for normal affinity erythrocytes (25.6 Torr) was significantly (p < 0.05) higher than that for high O2 affinity erythrocytes (15.9 Torr). Nevertheless, the VO2 for normal and high O2 affinity erythrocytes was approximately the same value, 6.5 ml.min-1.100 g-1 (p > 0.05). The O2 extraction rate was approximately 80% in normal and high O2 affinity erythrocytes. These results indicate that VO2 at low flow was independent of PVO2 in maximally contracting skeletal muscle, although PVO2 was reversely correlated with O2 affinity. We suggest that this PVO2-independency of VO2 at low flow is caused by long red-cell transit time in the capillary without diffusion-limitation and/or a 20% constant arterio-venous shunt of arterial O2 delivery.

Occurence of a sialylglycopeptide and free sialylglycans in hen's egg yolk.

Free sialylglycans (FSGs) and a sialylglycopeptide (SGP) as components of hen's egg yolk were found and their chemical structures were determined. SGP and FSGs were isolated from fresh egg yolk by treatment with phenol, gel filtration and successive chromatographies on columns of anion- and cation-exchangers. They were localized in the yolk plasma. The glycan moiety of SGP, which was liberated by PNGase digestion, was studied for the chemical structure by HPLC mapping with p-aminobenzoic ethylester-derivatization, sugar composition analysis, 1H nuclear magnetic resonance and matrix-assisted laser desorption/ionization time-of-flight (MALDI-TOF) mass spectrometry and the glycomoiety was found to be an N-linked disialyl-biantennary glycan. The amino acid sequence of the peptide moiety of SGP was determined to consist of Lys-Val-Ala-Asn-Lys-Thr, the Asn of which is modified with the disialylglycan moiety. FSGs were determined to be two free disialyl-biantennary glycans whose reducing end was either Man beta1-4GlcNAc (FSG-I) or Man beta1-4GlcNAc beta1-4GlcNAc (FSG-II). Since the molar value of SGP present in one egg yolk (2.8 micromol) is comparable to those of well-known major yolk proteins, low density lipoprotein, lipovitellins and phosvitin, it can be considered that SGP is one of the major components in hen's egg yolk.

Effect of blood blow and haematocrit on the relationship between muscle venous PO2 and oxygen uptake in dog maximally contracting gastrocnemius in situ.

1. To clarify the limiting factors for peak VO2 (VO2,peak), we measured muscle venous PO2, (PVO2) and VO2,peak under various O2 delivery conditions (arterial O2 concentration x flow) via the alteration of blood flow and haematocrit (Hct) in anaesthetized dog gastrocnemius muscle (n = 11) stimulated by 1 Hz isometrically and tetanically. 2. Two levels of flow (high and moderate) were maintained by using a pump (150 and 100 mL/min per 100 g, respectively). Haematocrit was adjusted to 45 and 30% by isovolaemic haemodilution with dextran (MW 40,000). 3. The decrease in Hct induced a 31% decrease in O2 delivery (P < 0.05), a 9-12% decrease in PVO2, a 23% decrease in VO2,peak (P < 0.05) and a 12-14% increase in O2 extraction (VO2/O2 delivery) at both flow levels. The decrease in flow induced a 24-25% decrease in O2 delivery (P < 0.05), a 15-17% decrease in PVO2 (P < 0.05), a 9% decrease in VO2,peak (P < 0.05) and a 19-22% increase in O2 extraction (P < 0.05) at both Hct levels. 4. The results suggest that the limiting factor of VO/,peak, unrelated to O2 diffusion limitation (i.e. the change in the heterogeneity of VO2 to O2 delivery ratio among capillaries), due to lowering Hct may be different from that due to lowering blood flow in maximally contracting muscle.

Flow-dependent influence of high-O2-affinity erythrocytes on peak VO2 in exercising muscle in situ.

To evaluate the influence of a high-O2 affinity of the erythrocyte and of flow rate on muscle's ability to extract O2 and develop force, we perfused dog gastrocnemius contracting isometrically at 4 Hz with normal-O2-affinity perfusate or high-O2-affinity perfusate at high and moderate flows (200 and 100 ml . min-1 . 100g-1, respectively). High-O2-affinity perfusate was prepared by incubating human citrate-phosphate-dextrose-stored erythrocytes with buffered saline containing cyanate (4 degrees C, 18 h) and normal-affinity perfusate by storing 2,3-diphosphoglycerate-rejuvenated erythrocytes in the same solution without cyanate. PO2 when blood is half oxygenated was 30.6 Torr for normal perfusate and 18.1 Torr for high-affinity perfusate. During 4-Hz stimulation, the tension developed by the muscle increased incrementally (positive staircase) to reach a peak value after 1.2-1.6 min for the normal perfusate and 0.6-0.7 min for the high-affinity perfusate (P < 0.05). The rate of decline during the early fatigue (measured from the onset of tension decline to 3 min) with high-affinity perfusate was significantly faster than it was with normal perfusate (P < 0.05). These findings suggest that both the staircase effect and the early fatigue are related to O2 availability, which is restricted when erythrocytes have a high O2 affinity. The peak O2 uptake values measured at 3 and 5 min were significantly lower (by 14-24%) with high-affinity perfusate than with normal perfusate at a given level of O2 delivery (arterial O2 content x flow) (P < 0.05). PO2 of venous effluent was proportionally related to peak O2 uptake. The present results indicate that neither blood flow nor O2 delivery is the sole determinant of the muscle's ability to extract O2.

Mixed disulphide formation in myoglobin of mouse (Mus musculus).

The myoglobin, isolated from murine skeletal muscles by chromatofocusing, showed the three components, one major and two minor, with different electrophoretic mobilities. The major component with the isoelectric point (pI) of 7.55 had one reactive SH/mole, while the others with pI of 7.32 and 7.16 showed none, which could be rendered fully reactive by treating the proteins with beta-mercaptoethanol. The three components were the same in their molecular weight (18 kDa), amino-acid composition with one Cys residue and oxygenation properties. By a sensitive high-performance liquid chromatography method, the occurrence of cysteine or glutathione mixed disulphide was verified in the two minor components. We conclude from these results and incubation experiments with low-molecular-weight thiols that the two minors were derived from the major by a mixed disulphide formation with either cysteine or glutathione of the cysteinyl SH at the 66th sequence.

Correlation between a sandwich ELISA and an in-vitro bioassay for erythropoietin in human plasma.

A sandwich-type enzyme-linked immunosorbent assay (ELISA) for human erythropoietin (EPO) using two anti-EPO monoclonal antibodies has been compared with an in-vitro EPO bioassay based on CFU-E colony formation in fetal mouse liver cell cultures. In normal subjects and non-uraemic anaemic patients the plasma EPO values estimated with the ELISA correlated well with those by the bioassay, and also inversely with the values of blood Hb, PCV and RBC counts. Dose-response curves for plasma and standard recombinant human EPO in the ELISA were parallel to each other. These results further confirm the validity for the ELISA in measuring circulating human EPO.

Oxygen affinities (P50) of myoglobins from four vertebrate species (Canis familiaris, Rattus norvegicus, Mus musculus and Gallus domesticus) as determined by a kinetic and an equilibrium method.

Rate constants of the reaction with oxygen of myoglobins from four vertebrate species (Canis familiaris, Rattus norvegicus, Mus musculus and Gallus domesticus) and the isolated alpha A and beta A chains of human adult hemoglobin (HbA) were determined by the stopped-flow-spectrophotomeric method. Half-saturation oxygen pressure (P50) of the proteins calculated from the rate constants, assuming a simple bimolecular reaction model, agreed very well with those directly determined by oxygen equilibria. The proteins used were freshly prepared, and fully characterized by electrophoretic and ultracentrifugal analyses. Sulphydryl groups in the Hb chains were ascertained to be completely regenerated.

Erythropoietin response to acute hypobaric or anaemic hypoxia in gentamicin-administered rats.

In an attempt to assess the erythropoietin (Epo) production site(s) in rat kidney, Epo response to hypoxia and renal histopathological changes were studied in rats administered with graded doses of gentamicin. Male Sprague-Dawley rats of 9 to 11 weeks old were used. Following a 14-day subcutaneous administration (67.5 or 33.8 mg kg-1 day-1) of gentamicin, a nephrotoxic aminoglycoside, selective proximal tubular lesions were produced. These gentamicin-administered rats were compared with normal rats with respect to Epo response to hypoxia. Two different kinds of hypoxic load, either 0.35 atm hypobaric hypoxia (PIO2 = 46 torr) or acute anaemia (Ht: 29.3 +/- 0.2% and [Hb]: 9.7 +/- 0.3 g dl-1) by withdrawing of blood corresponding to 1-2% of body weight was used. During the hypoxic period of up to 48 h, the peak renal venous plasma Epo titres of 3.1 +/- 0.6 and 4.3 +/- 0.6 U ml-1 was observed at the 6th h in normal hypobaric hypoxic and anaemic rats, compared with the prehypoxic value of 0.7 +/- 0.1 U ml-1. The Epo titres then declined gradually. In the rats which were administered gentamicin, Epo response pattern was the same as that observed in the normal rats, but the peak value decreased significantly to 0.8 +/- 0.3 and 1.1 +/- 0.4 U ml-1 in hypoxic and anaemic rats (P < 0.05). Histological examination revealed the selective damage to renal proximal convoluted tubules. The Epo response was reduced by the tissue damages, and restoration of the gentamicin-induced tissue injury was accompanied with restored Epo response to hypoxia.(ABSTRACT TRUNCATED AT 250 WORDS)

High affinity of blood for oxygen reduces oxygen uptake in contracting canine gracilis muscle.

To clarify the influence of blood flow with high-oxygen (O2)-affinity blood on oxygen consumption (VO2) in submaximally exercising skeletal muscle, we perfused the isolated dog gracilis (n = 8) contracting under 1 Hz stimulation alternatively with normal and high-O2-affinity blood, with a constant arterial O2 content (Ca,O2) and varying perfusion rates. The average P50 (oxygen partial pressure (PO2) for half-saturation at pH 7.40, PCO2 of 40 mmHg at 37 degrees C) of the high-O2-affinity blood prepared by carbamylation was 15.5 mmHg, and that of the normal blood 33.7 mmHg. With normal blood perfusion, the average VO2 above 6 ml min-1 (100 g)-1 of O2 delivery (Ca,O2 x flow) was 4.38 ml min-1 (100 g)-1 (range 4.17-4.68 ml min-1 (100 g)-1, and VO2 at the O2 delivery range of 6-5 and 4-2.5 ml min-1 (100 g)-1 decreased to 3.96 and 2.43 ml min-1 (100 g)-1, respectively. The PO2 of venous effluent (Pv,O2) at the O2 delivery of 6 ml min-1 (100 g)-1 was 33 mmHg. With low-P50 blood perfusion, VO2 was significantly less than with normal blood, both below the O2 delivery level of 6 ml min-1 (100 g)-1 and above it, even in the fairly high O2 delivery range of 8.5-12 ml min-1 (100 g)-1 (P < 0.05). Thus, high blood flow did not compensate for the reduced VO2 caused by high-O2-affinity blood. At values of Pv,O2 less than 33 mmHg, VO2 with low-P50 blood was not significantly different from that with normal blood (P > 0.05). The reduced VO2 in submaximally exercising skeletal muscle might be due to a slower O2 dissociation from the high-O2-affinity red cells and to a limited O2 diffusion resulting from the lower Pv,O2 value (which reflects mean end-capillary PO2).